Datasets:

iNeil77

/

pseudo-mini-pile

like 4

. The variations of blood pressure (BP) during exercise are important when there is a doubt about the normality of the values or a possibility of future abnormality. This is particularly true in children and adolescents with borderline or variable values, whose capacity of cardiovascular adaptation to sport is to be evaluated or who have a family history of hypertension. This study was undertaken to establish the profile of blood pressure on exercise during bicycle ergometry with reference to the height of the child or adolescent. BP and heart rate were measured in 651 healthy children aged 5 to 18 years during stress testing by bicycle ergometry. The sample was divided according to sex and height (120-139, 140-159, 160-180 cm), and according to resting BP values. During simple progressive exercise, the work required is increased by 10 watts every 2 minutes for children less than 140 cm tall, and by 20 watts when over 140 cm tall. BP and heart rate were measured during the last 30 seconds of each stage. Heart rate and systolic BP increased progressively with work load; on the other hand, the diastolic pressure did not change on exercise. The regression equation and the slope of systolic BP with respect to heart rate are reported, and allow comparison of different groups according to height, sex and resting systolic BP. The increase in systolic BP was the same in all groups except for boys taller than 160 cm in whom it was significantly greater (p less than 0,01). Sex and resting BP had no significant effect on the slope of systolic BP on exercise. The same applied to exercise lasting more than 6 minutes. The mean values and standard deviation of systolic BP at a heart rate of 150/min were, for each height group in ascending order: 121.5 +/- 19, 129 +/- 22 and 151 +/- 26 mmHg for boys, and 118.5 +/- 26, 126 +/- 21 and 137 +/- 26 mmHg for girls. In order to simplify the interpretation of the results obtained individually, charts in the form of an ellipse were plotted containing the usual values (95% of a healthy population) of systolic BP on exercise at the heart rate of 150/min, for each sex. They were established with reference to resting systolic BP and height. The diagnostic and prognostic value of this test will have to be evaluated by comparison with values obtained in hypertensive children.

The McHenry County Sheriff’s Office and Lake in the Hills Police Department have canceled SWAT training that was scheduled to take place Wednesday at a vacant village-owned building. Lake in the Hills police said Monday that the training was canceled because the funeral for fallen McHenry County Sheriff's Deputy Jacob Keltner is set that day. Keltner was shot and killed Thursday while trying to serve a warrant to a known fugitive in Rockford. A SWAT team is composed of members from several local police departments and is activated to assist agencies during high-risk situations outside normal police procedures, such as barricaded suspects, hostage rescue, active shooters and high-risk search or arrest warrants.

Stars of Vanuatu love story dance along red carpet at premiere of ''Tanna'' at Venice Film Festival. Rough Cut - no reporter narration. ROUGH CUT (NO REPORTER NARRATION) The cast of "Tanna", a film about forbidden love set in the South Pacific island of Vanuatu, performed a well-appreciated song and dance routine ahead of the film's Venice Film Festival premiere. "Tanna" set in the South Pacific island of Vanuatu premiered in Venice on Tuesday (September 8). The film was made by Australian documentary-makers, Bentley Dean and Martin Butler, and based around the love of Wawa (Marie Wawa) and Dain (Mungau Dain). The pair are deeply in love with one another, but Wawa is placed under pressure to marry a man from a rival tribe in a symbolic attempt to bring the warring groups closer together. However, Wawa and Dain flee and try to live together in various parts of the island, while they are pursued by their own tribe and the rival Imedin group.

Robertson Davies and Music Robertson Davies once described himself as a Failed Musician, but the musical education that Davies received, the role that music played in his life, and the use he made of music and musicians in his novels show that this is true only in the narrow sense that he did not earn a living from music. A proficient melopoetic artist, weaving music and musicians into his literary works with consummate skill and artistry, he was also professionally active as a music critic and a librettist. The German composer E.T.A. Hoffmann is the only historical musician to be given his real name and appear as a speaking character in any of the novels, but many of the authors fictional musicians are based on real-life models, and despite his great love of music, Davies often portrayed musicians as uncouth reprobates. Music, which sometimes provides structural foundations for Davies literary works and other times serves as a leitmotif, is often a portal to unseen worlds, including the realm of the dead and of supernatural phenomena, or the domains of religious experience or creativity.

Role of Neoadjuvant Chemotherapy with Cyclophosphamide, Adriamycin, 5-Fluorouracil (Caf Regimen) in Down Staging in Breast Cancer Background: The aim of the study is to assess the tumor response to neoadjuvant chemotherapy with cyclophosphamide, adriamycin, 5-fluorouracial (CAF regimen) in terms of decrease in breast tumor size (partial or complete clinically).To assess clinically the axillary lymph node status after neoadjuvant chemotherapy (conversion from palpable to nonpalpable). Methods: Thirty female patients of breast cancer were studied for down staging with two cycles of CAF regimen given at interval of 21 days. After 21 days of second cycle patients staging noted for effects. Results: Thirty female patients of breast cancer were studied. Maximum no. of patients between 31-40 years, mean age 46 years and median age 45 years, youngest patients 18 years, oldest patients 70 years, 22 patients responded to chemotherapy, out of 22, 1 (3.3%) showed a complete clinical response, 21 (70%) partial clinical response. Pre-menopausal 9/13 (69.2%) and post menopausal 13/17 (76.4%) showed clinical response, statistically not significant difference (df=1, x=1.33, p>0.05). Change in tumor size 40.09±25.20 sq, cm mean size to 21.88±27.43 sq. cm after chemotherapy was highly significant change (t=6.242, p<0.001). Overall response to chemotherapy was 73.3%, in stage II-87.5%, stage IIIA-75% and stage IIIB-50%. The overall response to axillary lymph node was 56.6%, statistically highly significant (p<0.001). Main side effects nausea and vomiting (60%) and hair loss, 43.3%, but none necessitated stoppage of chemotherapy. As a consequence to primary chemotherapy, conservation surgery (lumpectomy with axillary clearance) could be done in 43.3% of patients.Conclusion: CAF Preoperative chemotherapy regime is a satisfactory modality of treatment for stage II and III breast cancer with positive response rate of 73.3%. The down staging thus obtained permits breast conservation surgery in 43.3% of patients. The chemotherapy regime is well accepted by patients. INTRODUCTION The management of breast cancer ranges from loco/regional control with modified radical mastectomy to multipronged approach of breast conservation surgical techniques. Breast conservation is possible only if the tumor size is reduced. Neo adjuvant chemotherapy is being used frequently before surgery in large and locally advanced breast cancers aiming diminution of primary tumor size and knocking out putative micrometastasis to improve survival. MATERIALS AND METHODS The study was conducted from May, 2005 to Nov 2007 in the Department of General Surgery in conjunction with Department of Radiotherapy, Guru Nanak Dev Hospital, attached to Government Medical College, Amritsar. Informed consent was taken from all the patients included in the study. Patient population: Thirty female patients with breast carcinoma, stage II and stage III (TNM according to standard AJCC 1997 staging) in were studied. Two cycles of CAF or FAC regimen Swenerton were administered preoperatively as follows: -Cyclophosphamide 500 mg/m 2 I.V. day 1. Adriamycin (Doxorubicin) 50 mg/m 2 I.V. day 1,5-Fluorouracil 500 mg/m 2 I.V. day 1 and 8. The cycle repeated after 21 days. Inclusion Criteria: a) T 1 /T 2 /T 3 with N 0 /N 1 and M 0 ; Section: Surgery b) Any T with N 2 /N 3 and M 0 ; c) T 3 /T 4 with any N and M 0. Exclusion Criteria: Patient unfit for chemotherapy; previous antitumor chemotherapy, inflammatory breast carcinoma, concurrent other malignancy, of distant metastases and cardiovascular impairment were excluded. Evaluation: a) A detailed history and systemic examination was carried out. The size of the breast lump of the patient was measured with Vernier calipers or measuring tape. Evaluation of response was carried out in terms of change in the TNM staging (before and after neo adjuvant chemotherapy). Down staging was evaluated after 21 days from completion of second dose of chemotherapy. The response was assessed as complete response, partial response, no change or progressive disease by standard criteria as Haywards et al. RESULTS Maximum number of patients fell in the age groups of 31-40 years. The mean age of study population was 46 years and median age was 45 years. The youngest patient was 18 years female and the oldest patient was a 70 years female. Out of 30 patients, 22 patients responded to neo adjuvant chemotherapy (73.3%). Out of 22 patients, complete response was shown by 1 (3.3%) patient while the remaining 21 (70%) patients showed partial response. Patients in age group 11-20 years and 41-50 years had maximally responded to chemotherapy (i.e. 100%), compared to those in age group 61-70 years (i.e. 33.3%). Out of 13 premenopausal patients 9 (69.2%) had clinical response to chemotherapy. In post menopausal group, out of 17 patients, only 13 (76.4%) patients had clinical response to chemotherapy. Difference between response with neo adjuvant chemotherapy among premenopausal and postmenopausal was found to be statistically not significant (d f = 1; x 2 = 1.33; p>0.05) Mean tumor size in present study before chemotherapy was 40.09+25.20 sq.cm (range: 6.82-147.00 sq.cm) and that after chemotherapy was 21.88+27.43 cm 2 (range: 0-127.40 cm 2 ). Change in tumor size after chemotherapy was found to be statistically highly significant (t = 6.242; p<0.001).Present study showed 87.50% response to chemotherapy in stage II tumor, 75% response in stage IIIA and 50% response in stage IIIB tumor. Overall response to chemotherapy was 73.3% in stage II and stage III patients. Statistically highly significant difference was observed in clinical response to stage II tumor compared to that of stage III tumor (p<0.001). Before neo adjuvant chemotherapy, twenty three patients were having clinically palpable auxiliary lymph nodes. Out of which 20 patients were having mobile lymph nodes (N 1 ) and 3 patients were having fixed axillary lymph nodes (N 2 ).After neo adjuvant chemotherapy, only ten patients were having mobile palpable lymph nodes (N 1 ). So overall response to axillary lymph nodes was 56.6%. Change in lymph nodes status from clinically palpable to non-palpable was found to be statistically highly significant (p<0.001). The main side effects observed were nausea and vomiting (60%) and hair loss (43.3%) in all patients but none necessitated discontinuation of chemotherapy. Subsequent to preoperative chemotherapy, conservative surgery (lumpectomy with axillary clearance and segmental mastectomy with axillary clearance) could be done in 43.3% of the patients. Following surgery, all patients were discharged after removal of stitches. None had postoperative complications after surgery and follow up was done for prognosis and disease free survival for 12 months. DISCUSSION In our study, out of 30 patients 22 have responded to neo adjuvant chemotherapy (73.3%). Out of 22 patients, complete clinical response was shown by one patient (3.3%) while the remaining patients (70%) showed partial clinical response. Different studies have shown clinical response range from 70% to 85% and clinically complete response 6.6 to 27%. Hence, clinical response in present study also fell in the same range, but complete clinical response is less than that of different studies. The reasons for lower result in clinical complete response may be:(A) Our study group was very small (i.e. 30 patients) compared to other studies.(B)We have given only two cycles of CAF regimen preoperatively compared to 2 to 5 cycles of chemotherapy in other studies. Stage III a T0N2M0 T1N2M0 T2N2M0 T3N1M0 T3N2M0 ---13 3 ---2 -6. Section: Surgery Any T N3M0 --Present study showed that patients in age groups 11-20 years and 41-50 years had maximally responded to chemotherapy (100%) compared to those in age group 61-70 years (33.3%). But the correlation between age of patients and change in tumor size after neo adjuvant chemotherapy was found to be statistically not significant (r= -0.13859; p>0.05). It showed that there exists a negative but not significant correlation between age and change in tumor size which implies that as the age increases, the change in tumor size with neo adjuvant chemotherapy decreases although not significantly. Different studies have also shown that age is no specific criterion for clinical response to chemotherapy. Mean tumor size in present study before chemotherapy was 40. One study showed that the response to chemotherapy decreased as the tumor size increased (tumor size <5cm 48% response, tumor size 5-10 cm 20% response and tumor size >10cm 4% response). Similarly a study concluded that tumor size < or = 2cm responded maximally to chemotherapy (p<0.001). Another study concluded that tumor size had marked prognostic significance following chemotherapy. So prognostic significance of tumor size in present study correlates with other studies. So results of our study also correlate with other studies. In present study, 23 patients were having clinically palpable axillary lymph nodes out of which 20 patients were having mobile lymph nodes (N 1 ) and 3 patients were having fixed axillary lymph nodes (N 2 ). After chemotherapy, only 10 patients were having mobile palpable lymph nodes (N 1 ). So overall response to axillary lymph nodes was 56.6% and change in lymph node status from clinically palpable to non palpable was found to be statistically highly significant (p<0.001). In one study, 170 patients of locally advanced breast cancer were treated with neoadjuvant chemotherapy and 63% patients had negative axillary lymph nodes after chemotherapy. Similarly, a study showed that metastatic lymph nodes were found to be a sovereign predictor of tumor response as well as relapse. Hence, clinical response to axillary lymph nodes after neoadjvuant chemotherapy in present study correlates with the result of other studies. The prognostic value is greatest in aggressive tumour subtypes. The pathological response to preoperative therapy exhibits a complex interaction between the regimen delivered, the pathological complete response improvement, and long-term outcome, is not well understood. Side effects of chemotherapy regimen: Patients had nausea and vomiting as most common side effects during chemotherapy (60% cases), was controlled by antiemetic and antacid drugs. Second most common side effect noted among patients was loss of hair (43.3%). All the 30 patients completed two cycles of neoadjuvant chemotherapy. Procedures undertaken after chemotherapy: As a result of down staging of tumor with neoadjuvant chemotherapy, breast conservation surgery was possible in 13 patients (43.3%). Following surgery, all patients were discharged after removal of stitches. None had post-operative complications. All patients received 5 cycles of chemotherapy after surgery and follow up was done for prognosis and disease free survival for 12 months. CONCLUSION Based upon this study, we conclude that preoperative chemotherapy is a satisfactory modality of treatment for stage II and stage III breast cancer with a positive response rate of 73.3%. The down staging thus obtained permits breast conservation

The diagnosis of diabetes mellitus Assessment of diabetes control The OaTT should not be used to assess the degree of diabetes control. While this statement might appear obvious (and unnecessary) some practitioners still order an OaTT for this purpose, or to see "if the diabetes is improving". Similarly, the glycosylated haemoglobin and serum glycosamine levels should not be used to diagnose diabetes. However, they have now become well established means of assessing control over a period of time."

Q: If the ocean vanished, how long and how badly would it stink? For no apparent reason (this is a song, not a story, so I'm pretty economical with exposition) nearly every tetrapod on Earth has vanished, and water is gone from the oceans. Just sort of hand-waved out of existence; too many clouds if it all evaporated. The viewpoint character, the only remaining tetrapod we know about, is sitting on the lawn of a hotel in coastal New England looking out to sea, or what used to be the sea. There's nothing but mud. The water went away, leaving fish, whales, a thousand thousand slimy things, etc. (What? Whales are tetrapods? Look, a guitar solo!) It's been six or eight weeks since what I'll call The Event. What would a former harbor smell like? I don't want to dwell on anything ghoulish or disgusting, but it's a key part of the scene. A: The ocean is responsible of absorbing a large quantity of the heat Earth receives from the sun, and stabilizing Earth's rotation and atmosphere. Since the Earth's temperatures would be drastically different between day and night, and the atmosphere would also be greatly disturbed by the sudden new topography of the surface, you could expect the rotting process of sea and land animals to be incredibly fast. I'd expect the air to be unbreathable (if this word exists) between 2-3 weeks from the Event. Your lone tetrapod survived six weeks. Air is bathed in death's perfume, but he doesn't care anymore.

Jack White has spoken about being influenced by rap music recently, also describing Nicki Minaj as “brilliant” in a new interview. The former White Stripes frontman released his third solo album ‘Boarding House Reach’ on March 23. It saw White work with hip-hop musicians who had previously played with the likes of Kendrick Lamar and Kanye West. Speaking in a new interview with Clash, White revealed that he’s been recently listening to a lot of Kanye, Minaj, A Tribe Called Quest and “’80s and ’90s hip-hop specifically”. White went on to discuss Minaj’s work, saying that he was “shocked” by some of her lyrics. This comes after White made headlines for saying that he doesn’t understand what DJ Khaled does. Khaled’s 2017 Rihanna collaboration ‘Wild Thoughts’ heavily sampled Santana’s 1999 classic ‘Maria Maria’ and White recently described it as “just Santana’s song in its entirety”.

Chinese five spice powder is a blend of spices (often containing more than 5 spices) with the warm musty aroma of anise, cinnamon, cloves and pepper. It’s a signature flavor of many Chinese dishes including Char Siu and Cantonese Roast Pork. The addition of maltose (also known as rice syrup) to the marinade creates glossy glaze as the chicken roasts. While I prefer using maltose for its neutral taste and viscous texture, if you can’t find it, a neutral tasting honey will work as well. By marinating the chicken in the sauce for a day or two it not only imbues a stunning mahogany shade to the chicken, it draws out excess liquid, giving the chicken a firm, almost duck-like texture. Serve this five spice chicken over a bed of rice with some sauteed greens. If you want some extra sauce to pour over the chicken and rice, you can boil the remaining marinade, skimming off any foam that floats to the top until it’s thick and glossy. Whisk together the dark soy sauce, hoisin sauce, maltose, oyster sauce, garlic and five spice powder in a small bowl until well combined. Pour this marinade over the chicken thighs, flipping them over several times to coat evenly and marinate for at least 24 hours, or up to 48 hours. When you're ready to roast the chicken, prepare a baking sheet lined with a wire rack, and preheat the oven to 390 degrees F (200 C). Adding a layer of aluminum foil between the rack and baking sheet will make cleanup easier as the sugar in the marinade tends to burn to the pan. Remove the chicken from the marinade and place on the rack, skin-side up. Reserve the marinade for basting. Roast the chicken for 6 minutes, and then brush the tops with some of the remaining marinade. Continue roasting for another 6-8 minutes, or until the edges are lightly charred and the chicken registers 160 degrees F on an instant read thermometer. Remove the chicken from the oven and let it rest for 10 minutes before slicing and serving on top of rice.

Use of a Cell Transformation Assay with Established Cell Lines, and a Metabolic Cooperation Assay with V79 Cells for the Detection of Tumour Promoters: A Review Extensive studies on the safety evaluation of chemicals have indicated that a considerable number of non-genotoxic chemicals are carcinogenic. Tumour promoters are likely to be among these non-genotoxic carcinogens, and their detection is considered to be an important approach to the prevention of cancer. In this review, the results are summarised for in vitro transformation assays involving established cell lines, and for an assay for inhibition of gap junctional intercellular communication for the detection of tumour promoters, which involves V79 cells. Although the number of chemicals examined is still too small to permit a full evaluation of the correlation between in vitro cell transformation and in vivo carcinogenic-ity, it is clear that the sensitivity of the focus formation assay is very high. In the case of the metabolic cooperation assay, the sensitivity appears to be rather poor, but the assay can be considered to be useful because of its simple procedure and its considerable database. These in vitro assays for tumour promoters are recommended as useful tools for the detection of non-genotoxic carcinogens.

Synthesis and Biological Evaluation of Cyclic Analogues from Nitrone LQB-278: A New Potential Antileukemia Compound Background/Aim: A new set of LQB-nitrones and analogues was synthesized to evaluate anticancer activity based on the substitution of the terpenyl moiety of the antileukemic compound LQB-278 by the conformationally restricted cinnamyl ether. Materials and Methods: A structure-activity relationship study was performed in vitro on Jurkat cells to screen the antileukemic activity of LQB-nitrones and analogues and elucidate the mechanisms of action of the most active derivatives. Results: The cynamyl ramification and its ortho position aldehyde substitution improved the antileukemic activity. Three compounds showed an in vitro antiproliferative action, but only 5b induced apoptosis. Analysis of the molecular mechanisms showed increased expression of the cell cycle inhibitor p21CIP1/WAF1/Sdi1, caspase 3, Fas receptor, and Bax/Bcl-2 ratio. Conclusion: The cinnamyl derivative 5b (LQB-461) presented higher antileukemic effects than the prototype terpenyl nitrone, inducing Jurkat cell death by activating both extrinsic and intrinsic pathways of apoptosis. Therefore, this compound is a new promising candidate drug against leukemia.

Compaction of RFID Devices Using Data Compression When a coding system used to represent letters or numbers in transmitting messages has many unused codewords, it will be more than reasonable to compress the messages. Electronic product code (EPC) which is widely used by many RFID devices has plentiful unused codewords. As a matter of fact, the unused codewords are most by far of the codewords. In this paper, we check the suitableness of a commonly used compression algorithm for EPC of RFID devices.

Slavery in ancient Rome Origins In his Institutiones (161 AD), the Roman jurist Gaius wrote that: [Slavery is] the state that is recognized by the ius gentium in which someone is subject to the dominion of another person contrary to nature. — Gaius, Institutiones 1.3.2 The 1st century BC Greek historian Dionysius of Halicarnassus indicates that the Roman institution of slavery began with the legendary founder Romulus giving Roman fathers the right to sell their own children into slavery, and kept growing with the expansion of the Roman state. Slave ownership was most widespread throughout the Roman citizenry from the Second Punic War (218–201 BC) to the 4th century AD. The Greek geographer Strabo (1st century AD) records how an enormous slave trade resulted from the collapse of the Seleucid Empire (100–63 BC). The Twelve Tables, Rome's oldest legal code, has brief references to slavery, indicating that the institution was of long standing. In the tripartite division of law by the jurist Ulpian (2nd century AD), slavery was an aspect of the ius gentium, the customary international law held in common among all peoples (gentes). The "law of nations" was neither considered natural law, thought to exist in nature and govern animals as well as humans, nor civil law, belonging to the emerging bodies of laws specific to a people in Western societies. All human beings are born free (liberi) under natural law, but slavery was held to be a practice common to all nations, who might then have specific civil laws pertaining to slaves. In ancient warfare, the victor had the right under the ius gentium to enslave a defeated population; however, if a settlement had been reached through diplomatic negotiations or formal surrender, the people were by custom to be spared violence and enslavement. The ius gentium was not a legal code, and any force it had depended on "reasoned compliance with standards of international conduct." Vernae (singular verna) were slaves born within a household (familia) or on a family farm or agricultural estate (villa). There was a stronger social obligation to care for vernae, whose epitaphs sometimes identify them as such, and at times they would have been the children of free males of the household. The general Latin word for slave was servus. Slavery and warfare Throughout the Roman period many slaves for the Roman market were acquired through warfare. Many captives were either brought back as war booty or sold to traders, and ancient sources cite anywhere from hundreds to tens of thousands of such slaves captured in each war. These wars included every major war of conquest from the Monarchical period to the Imperial period, as well as the Social and Samnite Wars. The prisoners taken or re-taken after the three Roman Servile Wars (135–132, 104–100, and 73–71 BC, respectively) also contributed to the slave supply. While warfare during the Republic provided the largest figures for captives, warfare continued to produce slaves for Rome throughout the imperial period. Piracy has a long history of adding to the slave trade, and the period of the Roman Republic was no different. Piracy was particularly affluent in Cilicia where pirates operated with impunity from a number of strongholds. Pompey was credited with effectively eradicating piracy from the Mediterranean in 67 BC. Although large scale piracy was curbed under Pompey and controlled under the Roman Empire, it remained a steady institution and kidnapping through piracy continued to contribute to the Roman slave supply. Augustine lamented the wide scale practice of kidnapping in North Africa in the early 5th century AD. Trade and economy During the period of Roman imperial expansion, the increase in wealth amongst the Roman elite and the substantial growth of slavery transformed the economy. Although the economy was dependent on slavery, Rome was not the most slave-dependent culture in history. Among the Spartans, for instance, the slave class of helots outnumbered the free by about seven to one, according to Herodotus. In any case, the overall role of slavery in Roman economy is a discussed issue among scholars. Delos in the eastern Mediterranean was made a free port in 166 BC and became one of the main market venues for slaves. Multitudes of slaves who found their way to Italy were purchased by wealthy landowners in need of large numbers of slaves to labor on their estates. Historian Keith Hopkins noted that it was land investment and agricultural production which generated great wealth in Italy, and considered that Rome's military conquests and the subsequent introduction of vast wealth and slaves into Italy had effects comparable to widespread and rapid technological innovations. Augustus imposed a 2 percent tax on the sale of slaves, estimated to generate annual revenues of about 5 million sesterces—a figure that indicates some 250,000 sales. The tax was increased to 4 percent by 43 AD. Slave markets seem to have existed in every city of the Empire, but outside Rome the major center was Ephesus. Demography Estimates for the prevalence of slavery in the Roman Empire vary. Estimates of the percentage of the population of Italy who were slaves range from 30 to 40 percent in the 1st century BC, upwards of two to three million slaves in Italy by the end of the 1st century BC, about 35% to 40% of Italy's population. For the empire as a whole during the period 260–425 AD, according to a study done by Kyle Harper, the slave population has been estimated at just under five million, representing 10–15% of the total population of 50–60 million+ inhabitants. An estimated 49% of all slaves were owned by the elite, who made up less than 1.5% of the empire's population. About half of all slaves worked in the countryside where they were a small percentage of the population except on some large agricultural, especially imperial, estates; the remainder the other half were a significant percentage 25% or more in towns and cities as domestics and workers in commercial enterprises and manufacturers. Roman slavery was not based on ideas of race. Slaves were drawn from all over Europe and the Mediterranean, including Gaul, Hispania, North Africa, Syria, Germany, Britannia, the Balkans, Greece, etc. Those from outside of Europe were predominantly of Greek descent, while the Jewish ones never fully assimilated into Roman society, remaining an identifiable minority. The slaves (especially the foreigners) had higher mortality rates and lower birth rates than natives, and were sometimes even subjected to mass expulsions. The averaged recorded age at death for the slaves of the city of Rome was extraordinarily low: seventeen and a half years (17.2 for males; 17.9 for females). By comparison, life expectancy at birth for the population as a whole was in the mid-twenties (36% percent of men could expect to reach the age of 62 and 27% of women if they succeeded in reaching the age of 10). The overall impact of slavery on the Italian genetics was insignificant though, because the slaves imported in Italy were native Europeans, mostly from Germanic and Slavic tribes and very few if any of them had extra European origin. This has been further confirmed by recent biochemical analysis of 166 skeletons from three non-elite imperial-era cemeteries in the vicinity of Rome (where the bulk of the slaves lived), which shows that only one individual definitely came from outside of Europe (North Africa), and another two possibly did, but results are inconclusive. In the rest of the Italian peninsula, the fraction of non-European slaves was definitively much lower than that. Auctions and sales New slaves were primarily acquired by wholesale dealers who followed the Roman armies. Many people who bought slaves wanted strong slaves, mostly men. Child slaves cost less than adults although other sources state their price as higher. Julius Caesar once sold the entire population of a conquered region in Gaul, no fewer than 53,000 people, to slave dealers on the spot. Within the empire, slaves were sold at public auction or sometimes in shops, or by private sale in the case of more valuable slaves. Slave dealing was overseen by the Roman fiscal officials called quaestors. Sometimes slaves stood on revolving stands, and around each slave for sale hung a type of plaque describing his or her origin, health, character, intelligence, education, and other information pertinent to purchasers. Prices varied with age and quality, with the most valuable slaves fetching high prices. Because the Romans wanted to know exactly what they were buying, slaves were presented naked. The dealer was required to take a slave back within six months if the slave had defects that were not manifest at the sale, or make good the buyer's loss. Slaves to be sold with no guarantee were made to wear a cap at the auction. Debt slavery Nexum was a debt bondage contract in the early Roman Republic. Within the Roman legal system, it was a form of mancipatio. Though the terms of the contract would vary, essentially a free man pledged himself as a bond slave (nexus) as surety for a loan. He might also hand over his son as collateral. Although the bondsman could expect to face humiliation and some abuse, as a legal citizen he was supposed to be exempt from corporal punishment. Nexum was abolished by the Lex Poetelia Papiria in 326 BC, in part to prevent abuses to the physical integrity of citizens who had fallen into debt bondage. Roman historians illuminated the abolition of nexum with a traditional story that varied in its particulars; basically, a nexus who was a handsome but upstanding youth suffered sexual harassment by the holder of the debt. In one version, the youth had gone into debt to pay for his father's funeral; in others, he had been handed over by his father. In all versions, he is presented as a model of virtue. Historical or not, the cautionary tale highlighted the incongruities of subjecting one free citizen to another's use, and the legal response was aimed at establishing the citizen's right to liberty (libertas), as distinguished from the slave or social outcast (infamis). Cicero considered the abolition of nexum primarily a political maneuver to appease the common people (plebs): the law was passed during the Conflict of the Orders, when plebeians were struggling to establish their rights in relation to the hereditary privileges of the patricians. Although nexum was abolished as a way to secure a loan, debt bondage might still result after a debtor defaulted. Types of work Slaves worked in a wide range of occupations that can be roughly divided into five categories: household or domestic, imperial or public, urban crafts and services, agriculture, and mining. Epitaphs record at least 55 different jobs a household slave might have, including barber, butler, cook, hairdresser, handmaid (ancilla), wash their master's clothes, wet nurse or nursery attendant, teacher, secretary, seamstress, accountant, and physician. A large elite household (a domus in town, or a villa in the countryside) might be supported by a staff of hundreds. The living conditions of slaves attached to a domus (the familia urbana), while inferior to those of the free persons they lived with, were sometimes superior to that of many free urban poor in Rome. Household slaves likely enjoyed the highest standard of living among Roman slaves, next to publicly owned slaves, who were not subject to the whims of a single master. Imperial slaves were those attached to the emperor's household, the familia Caesaris. In urban workplaces, the occupations of slaves included fullers, engravers, shoemakers, bakers, mule drivers, and prostitutes. Farm slaves (familia rustica) probably lived in more healthful conditions. Roman agricultural writers expect that the workforce of a farm will be mostly slaves, managed by a vilicus, who was often a slave himself. Slaves numbering in the tens of thousands were condemned to work in the mines or quarries, where conditions were notoriously brutal. Damnati in metallum ("those condemned to the mine") were convicts who lost their freedom as citizens (libertas), forfeited their property (bona) to the state, and became servi poenae, slaves as a legal penalty. Their status under the law was different from that of other slaves; they could not buy their freedom, be sold, or be set free. They were expected to live and die in the mines. Imperial slaves and freedmen (the familia Caesaris) worked in mine administration and management. In the Late Republic, about half the gladiators who fought in Roman arenas were slaves, though the most skilled were often free volunteers. Successful gladiators were occasionally rewarded with freedom. However gladiators, being trained warriors and having access to weapons, were potentially the most dangerous slaves. At an earlier time, many gladiators had been soldiers taken captive in war. Spartacus, who led the great slave rebellion of 73–71 BC, was a rebel gladiator. Servus publicus A servus publicus was a slave owned not by a private individual, but by the Roman people. Public slaves worked in temples and other public buildings both in Rome and in the municipalities. Most performed general, basic tasks as servants to the College of Pontiffs, magistrates, and other officials. Some well-qualified public slaves did skilled office work such as accounting and secretarial services. They were permitted to earn money for their personal use. Because they had an opportunity to prove their merit, they could acquire a reputation and influence, and were sometimes deemed eligible for manumission. During the Republic, a public slave could be freed by a magistrate's declaration, with the prior authorization of the senate; in the Imperial era, liberty would be granted by the emperor. Municipal public slaves could be freed by the municipal council. Treatment and legal status According to Marcel Mauss, in Roman times the persona gradually became "synonymous with the true nature of the individual" but "the slave was excluded from it. servus non habet personam ('a slave has no persona'). He has no personality. He does not own his body; he has no ancestors, no name, no cognomen, no goods of his own." The testimony of a slave could not be accepted in a court of law unless the slave was tortured—a practice based on the belief that slaves in a position to be privy to their masters' affairs would be too virtuously loyal to reveal damaging evidence unless coerced. Rome differed from Greek city-states in allowing freed slaves to become citizens. After manumission, a male slave who had belonged to a Roman citizen enjoyed not only passive freedom from ownership, but active political freedom (libertas), including the right to vote. A slave who had acquired libertas was thus a libertus ("freed person", feminine liberta) in relation to his former master, who then became his patron (patronus). As a social class, freed slaves were libertini, though later writers used the terms libertus and libertinus interchangeably. Libertini were not entitled to hold public office or state priesthoods, nor could they achieve senatorial rank. During the early Empire, however, freedmen held key positions in the government bureaucracy, so much so that Hadrian limited their participation by law. Any future children of a freedman would be born free, with full rights of citizenship. Although in general freed slaves could become citizens, with the right to vote if they were male, those categorized as dediticii suffered permanent disbarment from citizenship. The dediticii were mainly slaves whose masters had felt compelled to punish them for serious misconduct by placing them in chains, branding them, torturing them to confess a crime, imprisoning them or sending them involuntarily to a gladiatorial school (ludus), or condemning them to fight with gladiator or wild beasts (their subsequent status was obviously a concern only to those who survived). Dediticii were regarded as a threat to society, regardless of whether their master's punishments had been justified, and if they came within a hundred miles of Rome, they were subject to reenslavement. Roman slaves could hold property which, despite the fact that it belonged to their masters, they were allowed to use as if it were their own. Skilled or educated slaves were allowed to earn their own money, and might hope to save enough to buy their freedom. Such slaves were often freed by the terms of their master's will, or for services rendered. A notable example of a high-status slave was Tiro, the secretary of Cicero. Tiro was freed before his master's death, and was successful enough to retire on his own country estate, where he died at the age of 99. However, the master could arrange that slaves would only have enough money to buy their freedom when they were too old to work. They could then use the money to buy a new young slave while the old slave, unable to work, would be forced to rely on charity to stay alive. Several emperors began to grant more rights to slaves as the empire grew. Claudius announced that if a slave was abandoned by his master, he became free. Nero granted slaves the right to complain against their masters in a court. And under Antoninus Pius, a master who killed a slave without just cause could be tried for homicide. Legal protection of slaves continued to grow as the empire expanded. It became common throughout the mid to late 2nd century AD to allow slaves to complain of cruel or unfair treatment by their owners. Attitudes changed in part because of the influence among the educated elite of the Stoics, whose egalitarian views of humanity extended to slaves. There are reports of abuse of slaves by Romans, but there is little information to indicate how widespread such harsh treatment was. Cato the Elder was recorded as expelling his old or sick slaves from his house. Seneca held the view that a slave who was treated well would perform a better job than a poorly treated slave. As most slaves in the Roman world could easily blend into the population if they escaped, it was normal for the masters to discourage slaves from running away by putting a tattoo reading "Stop me! I am a runaway!" or "tax paid" if the slaves were owned by the Roman state on the foreheads of their slaves. For this reason, slaves usually wore headbands to cover up their disfiguring tattoos and at the Temple of Asclepius, the Greek god of healing, in Ephesus, archeologists have found thousands of tablets from escaped slaves asking Asclepius to make their tattoos on their foreheads disappear. Crucifixion was the capital punishment meted out specifically to slaves, traitors, and bandits. Marcus Crassus was supposed to have concluded his victory over Spartacus in the Third Servile War by crucifying 6,000 of the slave rebels along the Appian Way. Rebellions and runaways Moses Finley remarked, "fugitive slaves are almost an obsession in the sources". Rome forbade the harbouring of fugitive slaves, and professional slave-catchers were hired to hunt down runaways. Advertisements were posted with precise descriptions of escaped slaves, and offered rewards. If caught, fugitives could be punished by being whipped, burnt with iron, or killed. Those who lived were branded on the forehead with the letters FUG, for fugitivus. Sometimes slaves had a metal collar riveted around the neck. One such collar is preserved at Rome and states in Latin, "I have run away. Catch me. If you take me back to my master Zoninus, you'll be rewarded." There was a constant danger of servile insurrection, which had more than once seriously threatened the republic. The 1st century BC Greek historian Diodorus Siculus wrote that slaves sometimes banded together to plot revolt. He chronicled the three major slave rebellions: in 135–132 BC (the First Servile War), in 104–100 BC (the Second Servile War), and in 73–71 BC (the Third Servile War). Serfdom In addition to slavery, the Romans also practiced serfdom. By the 3rd century AD, the Roman Empire faced a labour shortage. Large Roman landowners increasingly relied on Roman freemen, acting as tenant farmers, instead of slaves to provide labour. The status of these tenant farmers, eventually known as coloni, steadily eroded. Because the tax system implemented by Diocletian assessed taxes based on both land and the inhabitants of that land, it became administratively inconvenient for peasants to leave the land where they were counted in the census. In 332 AD Emperor Constantine issued legislation that greatly restricted the rights of the coloni and tied them to the land. Some see these laws as the beginning of medieval serfdom in Europe. Classical Roman religion The religious holiday most famously celebrated by slaves at Rome was the Saturnalia, a December festival of role reversals during which time slaves enjoyed a rich banquet, gambling, free speech and other forms of license not normally available to them. To mark their temporary freedom, they wore the pilleus, the cap of freedom, as did free citizens, who normally went about bareheaded. Some ancient sources suggest that master and slave dined together, while others indicate that the slaves feasted first, or that the masters actually served the food. The practice may have varied over time. Macrobius (5th century AD) describes the occasion thus: Meanwhile the head of the slave household, whose responsibility it was to offer sacrifice to the Penates, to manage the provisions and to direct the activities of the domestic servants, came to tell his master that the household had feasted according to the annual ritual custom. For at this festival, in houses that keep to proper religious usage, they first of all honor the slaves with a dinner prepared as if for the master; and only afterwards is the table set again for the head of the household. So, then, the chief slave came in to announce the time of dinner and to summon the masters to the table. Saturnalian license also permitted slaves to enjoy a pretense of disrespect for their masters, and exempted them from punishment. The Augustan poet Horace calls their freedom of speech "December liberty" (libertas Decembri). In two satires set during the Saturnalia, Horace portrays a slave as offering sharp criticism to his master. But everyone knew that the leveling of the social hierarchy was temporary and had limits; no social norms were ultimately threatened, because the holiday would end. Another slaves' holiday (servorum dies festus) was held August 13 in honor of Servius Tullius, the legendary sixth king of Rome who was the child of a slave woman. Like the Saturnalia, the holiday involved a role reversal: the matron of the household washed the heads of her slaves, as well as her own. The temple of Feronia at Terracina in Latium was the site of special ceremonies pertaining to manumission. The goddess was identified with Libertas, the personification of liberty, and was a tutelary goddess of freedmen (dea libertorum). A stone at her temple was inscribed "let deserving slaves sit down so that they may stand up free." Female slaves and religion At the Matralia, a women's festival held June 11 in connection with the goddess Mater Matuta, free women ceremonially beat a slave girl and drove her from the community. Slave women were otherwise forbidden participation. Slave women were honored at the Ancillarum Feriae on July 7. The holiday is explained as commemorating the service rendered to Rome by a group of ancillae (female slaves or "handmaids") during the war with the Fidenates in the late 4th century BC. Weakened by the Gallic sack of Rome in 390 BC, the Romans next had suffered a stinging defeat by the Fidenates, who demanded that they hand over their wives and virgin daughters as hostages to secure a peace. A handmaid named either Philotis or Tutula came up with a plan to deceive the enemy: the ancillae would put on the apparel of the free women, spend one night in the enemy camp, and send a signal to the Romans about the most advantageous time to launch a counterattack. Although the historicity of the underlying tale may be doubtful, it indicates that the Romans thought they had already had a significant slave population before the Punic Wars. Mystery cults The Mithraic mysteries were open to slaves and freedmen, and at some cult sites most or all votive offerings are made by slaves, sometimes for the sake of their masters' wellbeing. The cult of Mithras, which valued submission to authority and promotion through a hierarchy, was in harmony with the structure of Roman society, and thus the participation of slaves posed no threat to social order. Stoic philosophy The Stoics taught that all men were manifestations of the same universal spirit, and thus by nature equal. Stoicism also held that external circumstances (such as being enslaved) did not truly impede a person from practicing the Stoic ideal of inner self-mastery: It has been said that one of the more important Roman stoics, Epictetus, spent his youth as a slave. Early Christianity Both the Stoics and some early Christians opposed the ill-treatment of slaves, rather than slavery itself. Advocates of these philosophies saw them as ways to live within human societies as they were, rather than to overthrow entrenched institutions. In the Christian scriptures, equal pay and fair treatment of slaves was enjoined upon slave masters, and slaves were advised to obey their earthly masters, even if their masters are unfair, and lawfully obtain freedom if possible. Certain senior Christian leaders (such as Gregory of Nyssa and John Chrysostom) called for good treatment for slaves and condemned slavery, while others supported it. Christianity gave slaves an equal place within the religion, allowing them to participate in the liturgy. According to tradition, Pope Clement I (term c. 92–99), Pope Pius I (158–167) and Pope Callixtus I (c. 217–222) were former slaves. In literature Although ancient authors rarely discussed slavery in terms of morals, because their society did not view slavery as the moral dilemma we do today, they included slaves and the treatment of slaves in works in order to shed light on other topics—history, economy, an individual's character—or to entertain and amuse. Texts mentioning slaves include histories, personal letters, dramas, and satires, including Petronius' Banquet of Trimalchio, in which the eponymous freedman asserts "Slaves too are men. The milk they have drunk is just the same even if an evil fate has oppressed them." Many literary works may have served to help educated Roman slave owners navigate acceptability in the master-slave relationships in terms of slaves' behavior and punishment. To achieve this navigation of acceptability, works often focus on extreme cases, such as the crucifixion of hundreds of slaves for the murder of their master. We must be careful to recognize these instances as exceptional and yet recognize that the underlying problems must have concerned the authors and audiences. Examining the literary sources that mention ancient slavery can reveal both the context for and contemporary views of the institution. The following examples provide a sampling of different genres and portrayals. Plutarch Plutarch mentioned slavery in his biographical history in order to pass judgement on men's characters. In his Life of Cato the Elder, Plutarch revealed contrasting views of slaves. He wrote that Cato, known for his stringency, would resell his old servants because "no useless servants were fed in his house," but that he himself believes that "it marks an over-rigid temper for a man to take the work out of his servants as out of brute beasts." Cicero A prolific letter writer, Cicero even wrote letters to one of his administrative slaves, one Marcus Tullius Tiro. Even though Cicero himself remarked that he only wrote to Tiro "for the sake of keeping to [his] established practice," he occasionally revealed personal care and concern for his slave. Indeed, just the fact that Tiro had enough education and freedom to express his opinions in letters to his master is exceptional and only allowed through his unique circumstances. First, as an administrative slave, Tiro would have enjoyed better living and working conditions than the majority of slaves working in the fields, mines, or workhouses. Also, Cicero was an exceptional owner, even taking Tiro's education into his own hands. While these letters suggest a familiarity and connection between master and slave, each letter still contains a direct command, suggesting that Cicero calculatingly used familiarity in order to ensure performance and loyalty from Tiro. Roman comedies In Roman comedy, servi or slaves make up the majority of the stock characters, and generally fall into two basic categories: loyal slaves and tricksters. Loyal slaves often help their master in their plan to woo or obtain a lover (the most popular plot-driving element in Roman comedy). They are often dim, timid, and worried about what punishments may befall them. Trickster slaves are more numerous and often use their masters' unfortunate situation to create a "topsy-turvy" world in which they are the masters and their masters are subservient to them. The master will often ask the slave for a favor and the slave only complies once the master has made it clear that the slave is in charge, beseeching him and calling him lord, sometimes even a god. These slaves are threatened with numerous punishments for their treachery, but always escape the fulfillment of these threats through their wit. Depictions of slaves in Roman comedies can be seen in the work of Plautus and Publius Terentius Afer. Dartmouth associate professor Roberta Stewart has stated that Plautus’ plays represent slavery "as a complex institution that raised perplexing problems in human relationships involving masters and slaves.” Terence added a new element to how slaves were portrayed in his plays, due to his personal background as a former slave. In the work Andria, slaves are at the centerpiece off the plot. In this play, Simo, a wealthy Athenian wants his son, Pamphilius, to marry one girl but Pamphilius has his sights set on another. Much of the conflict in this play revolves around schemes with Pamphilius's slave, Davos, and the rest of the characters in the story. Many times throughout the play, slaves are allowed to engage in activity, such as the inner and personal lives of their owners, that wouldn't normally be seen with slaves in every day society. This is a form of satire by Terence due to the unrealistic nature of events that occurs between slaves and citizens in his plays. Emancipation Freeing a slave was called manumissio, which literally means "sending out from the hand". The freeing of the slave was a public ceremony, performed before some sort of public official, usually a judge. The owner touched the slave on the head with a staff and he was free to go. Simpler methods were sometimes used, usually with the owner proclaiming a slave's freedom in front of friends and family, or just a simple invitation to recline with the family at dinner. A felt cap called the Pileus was given to the former slave as symbol of manumission. Slaves were freed for a variety of reasons; for a particularly good deed toward the slave's owner, or out of friendship or respect. Sometimes, a slave who had enough money could buy his freedom and the freedom of a fellow slave, frequently a spouse. However, few slaves had enough money to do so, and many slaves were not allowed to hold money. Slaves were also freed through testamentary manumission, by a provision in an owner's will at his death. Augustus restricted such manumissions to at most a hundred slaves, and fewer in a small household. Already educated or experienced slaves were freed the most often. Eventually the practice became so common that Augustus decreed that no Roman slave could be freed before age 30. Freedmen A freed slave was the libertus of his former master, who became his patron (patronus). The two had mutual obligations to each other within the traditional patronage network. The terms of his manumission might specify the services a libertus owed. A freedman could "network" with other patrons as well. As a social class, former slaves were libertini. Men could vote and participate in politics, with some limitations. They could not run for office, nor be admitted to the senatorial class. The children of former slaves enjoyed the full privileges of Roman citizenship without restrictions. The Latin poet Horace was the son of a freedman, and an officer in the army of Marcus Junius Brutus. Some freedmen became very powerful. Many freedmen had important roles in the Roman government. Freedmen of the Imperial families often were the main functionaries in the Imperial administration. Some rose to positions of great influence, such as Narcissus, a former slave of the Emperor Claudius. Other freedmen became wealthy. The brothers who owned House of the Vettii, one of the biggest and most magnificent houses in Pompeii, are thought to have been freedmen. A freedman designed the amphitheater in Pompeii. A freedman who became rich and influential might still be looked down on by the traditional aristocracy as a vulgar nouveau riche. Trimalchio, a character in the Satyricon, is a caricature of such a freedman.

Knowledge-Based Flow of Control in Computer-Aided Learning In this paper I examine the utilisation of knowledge representation in Computer-Aided Learning (CAL) with the aim of establishing knowledge-based CAL techniques that are best suited to current technology. Most existing knowledge-based CAL systems attempt to generate the entire instructional sequence directly from a domain knowledge base. Such systems suffer from several limitations. These limitations include: 1. It is questionable whether the techniques exist to produce such systems for any but a highly restricted set of domains. 2. Even for those domains in which such systems can be produced the overheads are prohibitive for most purposes. Given these limitations, I argue that knowledge representation should be utilised in CAL only for those aspects of the instructional process for which it results in substantial gains without prohibitive overheads. I demonstrate that one aspect of CAL for which this holds is for managing flow of control within instructional material. I provide a detailed description of feature networks. These are a variant of M.A.K. Hallidays system network formalism. Feature networks are a knowledge representation formalism that efficiently encodes exactly the knowledge that is required for knowledge-based flow of control. It is shown that computer based lessons that utilise feature networks for control flow of control are extremely economic in terms of both authoring time and computer resources while providing highly responsive tuition. DABIS, a system that embodies the methodology outlined above, has been implemented and is described.

What's the best way to identify knockoff fashion from designer items? What’s the Best Way to Identify Knockoffs From Designer Items? Fake designer bags at a Hong Kong market in 2007. Fakes are harder and harder to differentiate from the real ones, as production houses that produce the real ones are starting to “knock off” the very work that they do for expensive luxury labels. However, the best way to recognize fakes is to look at the details. Listed below are five good details to look for when distinguishing a fake from a real product. Stitching on fakes is sloppily done. The lines are not clean, the threads are not trimmed, and often times the seams are not sewn straight. The quality of the fabric or leather will be compromised in fakes, because as you know, the way to get costs down is to use a cheaper alternative. If you can produce a fake in China, it’s way cheaper than going to a mill in Italy to have something custom made. Hence the fakes will feel cheaper to your touch. Again, you get what you pay for. Zippers and buttons or findings will be of substandard quality. Buttons will feel lighter if they are made of plastic-coated in metal instead of real metal. Zippers will come off the tracks or lose teeth prematurely if they are fake. The goodies. The real bags and shoes always come with the dust bags, the box, even ID cards for certain designers. Cheap imitations will not have these, or if they do, even the dust bags will have a cheap feel to them. Don’t get caught trying to buy fakes abroad. It is actually illegal in some countries, and rumor has it you could get caught at customs—not a great way to make your way back to the U.S. at all. Fashion and Style: Does watching the Victoria’s Secret fashion show make women feel inadequate? Fashion Trends: Why is Lululemon so popular? Fashion and Style: What is the difference between made to measure and bespoke?

Factors affecting the occurrence of dark-cutting beef and selected carcass traits in finished beef cattle. A data set was used to determine how various factors affect the occurrence of dark-cutting beef and selected carcass traits in finished beef cattle. Data were collected in 1989 and 1990 from one packer with plants located in Amarillo, TX; Boise, ID; Dakota City, NE; and Garden City, KS. The data set consisted of 3,659 lots consisting of 724,639 cattle. Compared with those at Boise and Dakota City, cattle slaughtered in Amarillo and Garden City had a higher incidence of dark cutters (1.1 vs.3%; P <.01) and a lower quality grade (50 vs 64% Choice plus Prime; P <.01). The highest incidences of dark cutters occurred during August, September, and October (1.1 to 1.4%; P <.01), with incidences of.4 to.7% during the other months. Carcass quality grade was higher during January, February, and March compared with May through November (60 to 62% Choice plus Prime vs 52 to 58%; P <.01). As the number of cattle in a lot increased, the incidence of dark cutters increased from.4 to 1.2% (P <.01), and quality grade declined from 62 to 52% Choice plus Prime (P <.01). As the mean weight of cattle in the lot increased, the incidence of dark cutters declined from.94 to.6% (P <.01), and carcasses grading Choice plus Prime increased from 56 to 62% (P <.01). With cattle held over a weekend or holiday, ("carry cattle") the incidence of dark cutters increased from.8 to 1.6% (P <.01). We conclude that packing plant location, month of the year, weight of cattle, carry cattle, and number of cattle in a lot are most likely to influence the incidence of dark cutters and carcass quality traits.

PORTLAND, Ore. (AP) Samuel Armenteros scored his first MLS goal in the 81st minute and the Portland Timbers won their fifth straight game with a 2-1 victory over Los Angeles FC on Saturday. Carlos Vela tied the match for LAFC in the 74th minute after Cristhian Paredes scored for the Timbers to open the second half. The Timbers (5-3-2) are 4-0 at home after starting the season winless for five straight matches on the road because of construction at Providence Park. Expansion LAFC (6-3-2) dropped just its third match on the road this year. The Timbers broke through with Paredes’ goal in the 52nd minute, which came on a rebound of Diego Valeri‘s free kick. It was the first MLS goal for the Paraguayan midfielder, who just turned 20 on Friday. Vela curled a shot over Timbers’ goalkeeper Jeff Antinella for his team-leading seventh goal of the season. It snapped Portland’s run of 353 minutes without conceding a goal. Armenteros, who was subbed in for Fanendo Adi in the 75th minute, scored some six minutes later and celebrated by running up the stairs behind the goal to the stands where he hugged Timbers Army supporters. Portland had three straight shutouts heading into the game, but they got hit in the ninth minute when defender Liam Ridgewell left the game with a non-contact injury. Although he was able to walk off the field unaided, he was replaced by Julio Cascante. Adi had a clear strike that caromed off the crossbar in the ninth minute for probably the best chance in the first half. Both teams are likely to see World Cup absences as early as next week. Officially, MLS teams don’t have to release players until May 28, although it’s likely they’ll let them go sooner in the interest of good relations with the national teams and their federations. The Timbers have two players that were on provisional World Cup rosters released earlier in the week. Midfielder David Guzman was on the roster for Costa Rica, which went ahead and named its World Cup squad, so he’s likely assured a spot in Russia. Midfilder Andy Polo was named to Peru’s provisional roster. LAFC’s players named to preliminary rosters include forward Carlos Vela (Mexico), forward Marco Urena (Costa Rica), Steve Beitashour (Iran) and defender/midfielder Omar Gaber (Egypt). Defender Laurent Ciman is expected to be on Belgium’s provisional roster, which will be announced Monday. Urena is recovering from facial surgery and was not available for LAFC on Saturday.

Acute syndesmosis injuries associated with ankle fractures: current perspectives in management. Ankle syndesmosis injuries frequently occur with ankle fractures, but their treatment remains controversial. Although specific clinical and radiographic diagnostic measures are generally well-accepted, there remains a lack of consensus with respect to the treatment of these injuries. Controversy arises at almost every phase of treatment including: type of fixation (screw size, type of implant), number of cortices required for fixation, and need for hardware removal. Regardless of fixation technique chosen, the most important goal should be anatomic reduction and restoration of the syndesmosis and ankle mortise as this is the only significant predictor of functional outcome.

Mathias Cormann defended Tony Abbott's "no cuts to the ABC and SBS" pre-election promise, saying the Prime Minister told the truth and they were not cuts but efficiency dividends. The Communications Minister on Wednesday detailed a $254 million funding cut to the ABC, which means the national broadcaster will receive $5.2 billion over four years, rather than $5.5 billion. The SBS will also have its budget cut by $25.2 million. Reduced funding: Financce Minister Mathias Cormann says the ABC is not having its funding cut, it is facing an efficiency dividend, Credit:Alex Ellinghausen When asked about Mr Abbott's pre-election comments, Senator Cormann said "well, they're not cuts". "The Prime Minister absolutely told the truth. We are not making cuts. What we are making sure happens with the ABC is what happens with every other taxpayer funded organisation across government and that is to ensure that it operates as efficiently as possible," he said.

Was it Right New York Knicks Coachs Decision to Recruit a Point Guard? The aims of this study was to analyze New York Knicks Coachs decision in recruiting a point guard. In the 2019-2020 season, NY Knicks ended up in ranked 25 of 30 NBA teams. NY Knicks is ranked 29th for FG% and 24th for 3PA and 3P%. NY Knicks need a shooter to strengthen the team. But interestingly, the coach recruited a point guard. In the first 25 games in 2020-2021 season without this point guard, NY Knicks recorded 11 wins and 14 loses and is in 9th place on the eastern conference. In the next 25 games after new point guard joined, NY Knicks recorded 14 wins and 11 loses and is in 7th position. This means that NY Knicks' game has increased and has the potential for the playoffs. The research method uses descriptive quantitative. The data source is taken from the NBA official website.

Glucose responsive neurons of the paraventricular thalamus control sucrose-seeking behavior Feeding behavior is governed by homeostatic needs and motivational drive to obtain palatable foods. Here, we identify a population of glutamatergic neurons in the paraventricular thalamus, which express the glucose transporter Glut2 (Scl2a2) and project to the nucleus accumbens. These neurons are activated by hypoglycemia and, in freely moving mice, their activation by optogenetics or Slc2a2 inactivation increases motivated sucrose but not saccharin-seeking behavior. These neurons may control sugar overconsumption in obesity and diabetes. remained similarly low in NG2KO mice and control mice (Suppl. Fig. 1a) indicating similar learning capacity. However, NG2KO mice performed better in this task (Fig. 1a,b). Next, we subjected mice to a progressive ratio (PR) schedule, which consisted of a systematic withinsession increase in the number of nosepokes required to earn each successive reward. NG2KO mice again performed better, with higher number of active nosepokes, of rewards obtained, and a higher maximal operant response (breakpoint) indicating higher motivation (Fig. 1c-e). Similar behavioral tasks were then performed with the non-caloric artificial sweetener saccharin. In FR1 (Fig. 1f,g) and PR (Fig. 1h-j) experiments NG2KO mice and control mice performed identically indicating that neuronal Glut2-dependent glucose sensing responds to sucrose and is insensitive to artificial sweeteners. Operant conditioning is associated with activity of the nucleus accumbens (NAc) 13, 14. We therefore investigated whether Glut2-positive neurons or axons could be observed in the NAc using Slc2a2-cre;Rosa26tdtomato mice 15. We observed numerous red fibers within the medial part of the NAc, mainly close to the anterior commissure and around the ventral end of the lateral ventricle (Suppl. Fig. 2a). To determine the origin of those fibers, we performed retrograde tracing experiments in which the cholera toxin B subunit-AlexaFluor 488 conjugate (CTxB) was injected bilaterally in the NAc of Slc2a2-cre;Rosa26tdtomato mice ( Fig. 2a and Suppl. Fig. 2b). Three weeks later, we screened the entire brain for tdtomato cell bodies labeled with CTxB. No colabeling with the rare Glut2-neurons present in the ventral tegmental area, the prefrontal cortex, the hippocampus or the different hypothalamic nuclei could be found. Co-labeled cells were found only in the paraventricular thalamic nucleus (PVT) (Fig. 2b) where 91 ± 1.7% of the PVT Glut2 neurons were also labeled with CTxB. Next, we assessed whether the firing activity of tdtomato positive neurons was responsive to changes in extracellular glucose concentrations (Fig. 2c). In the presence of 5 mM glucose these neurons showed a low basal activity, which was markedly enhanced in the presence of 0.5 mM glucose. Returning the glucose to 5 mM restored the initial firing activity (Fig. 2df). The same glucose-dependent increase in firing activity was seen in the presence a cocktail of synaptic inhibitors indicating that glucose-sensing is cell-autonomous (Suppl. Fig. 3a). When glycolysis was inhibited by 10 mM glucosamine in the presence of 5 mM glucose, firing was significantly increased (Fig. 2g-i). Fructose at 5 mM failed to suppress firing induced by 0.5 mM glucose (Suppl. Fig. 3b). Finally, patch clamp analysis of Glut2 neurons from mice previously injected with 2-deoxy-D-glucose showed that their firing frequency at 5 mM glucose was identical to that seen during exposure to 0.5 mM glucose of Glut2neurons from nave mice and did not change when glucose was lowered to 0.5 mM (Fig. 2jl). Tdtomato negative cells were also recorded in the presence of 5 or 0.5 mM glucose; they were found to be unresponsive to hypoglycemia (Suppl. Fig. 3c). Thus, PVT Glut2 neurons are activated by low glucose concentrations or by glycolysis inhibition; their firing rate cannot be suppressed by fructose, and they are responsive to in vivo induced neuroglucopenia. To investigate the nature of Glut2 projections to the NAc, Slc2a2-cre mice were injected in the PVT with an AAV-DIO-ChR2-eYFP 16 (Fig. 3a). Brain slices obtained three weeks later displayed numerous eYFP-positive neurons in the PVT and fibers in the NAc (Fig. 3b,c). Electrophysiological recordings from medium spiny neurons (MSNs) of the NAc together with local light-stimulation of the ChR2-expressing terminals (Fig. 3c) revealed postsynaptic inward currents (Fig. 3d), which could be totally abolished by tetrodotoxin, a blocker of action potential-dependent neurotransmitter release (Fig. 3d,e). The typical fast kinetics of those inward currents suggested they were -amino-3-hydroxy-5-methyl-4isoxazolepropionic receptors (AMPARs) excitatory post-synaptic currents (EPSCs). This was confirmed by the application of the AMPAR antagonist DNQX that entirely suppressed light-induced EPSCs in all the cells tested ( Fig. 3f-g). Thus, light-stimulation of NAcprojecting PVT Glut2-neurons triggers glutamate release onto NAc MSNs. To test the functional role of the PVT Glut2 excitatory inputs onto the NAc, we used an operant conditioning protocol associated with in vivo optogenetic activation of these neurons. Slc2a2-cre mice and control mice were injected in the PVT with an AAV-DIO-ChR2-eYFP and equipped with a fiber-optic cannula. Following recovery, mice were tested during operant conditioning to obtain sucrose. At the beginning of each behavioral session, mice were connected to a light source via an optical fiber and stimulated during the entire duration of the session. During the training/learning period (FR1 schedule), the ChR2expressing mice consumed significantly more reward (Fig. 3h) and performed more active nosepokes (Fig. 3i) than the control mice. Moreover, during the progressive ratio schedule mice expressing ChR2 showed increased motivated sucrose-seeking behavior by obtaining more rewards and showing increased breakpoint (Fig. 3j,k). To confirm that increased motivated behavior was light-dependent we tested the same mice in a progressive ratio schedule one month later without light stimulation. No difference in motivated behavior was seen between genotypes (Suppl. Fig. 4b,c). A similar experiment was repeated but with light stimulation through fiber-optic cannulas placed bilaterally at the NAc level. Light stimulation increased motivated sucrose seeking behavior as assessed in FR1 and progressive ratio schedules (Suppl. Fig. 5). When tested three days later without light stimulation, the breakpoint value of control mice remained the same and ChR2-expressing mice exhibited similar motivation as controls. Note that learning during the FR1 protocol was slower as compared to the previous experiment (Fig 3h,i), probably because these mice were implanted with two (instead of one) optical fibers, which may hamper access to the nosepoke holes. Finally, we inactivated Slc2a2 selectively in the PVT of Slc2a2 loxP/loxP mice by stereotactic injection of an AAV-Cre-GFP (Fig 3l). PCR analysis of genomic DNA extracted from the PVT and fluorescence microscopy analysis confirmed proper injection of the virus and recombination of Slc2a2 floxed allele in the AAV-Cre-GFP injected mice (Suppl. Fig. 6). These mice displayed increased motivation to get sucrose compared to control mice as reflected by the higher number of rewards and increased breakpoint (Fig. 3m,n). Collectively, our data show that Glut2-neurons from the PVT and projecting to the NAc form a population of hypoglycemia-activated, glutamatergic neurons, which induce EPSCs in MSN neurons. Their firing rate can be increased by glycolysis inhibition, but cannot be suppressed by fructose, indicating that they are controlled by glucose metabolism. When activated by hypoglycemia, optogenetics, or by Slc2a2 inactivation, they increase motivated sugar-feeding behavior indicating a dominant role in motivated sugar seeking. The possible relevance of these results to human physiology is suggested by the observations that mild hypoglycemia induces marked increase in synaptic activity in this part of the thalamus 17 and that variants in SLC2A2 are associated with increased intake of sugar-containing foods 18 and conversion from impaired glucose tolerance to type 2 diabetes 19. This newly identified neuronal pathway may, thus, represent a new target for the prevention of metabolic disease aiming at restoring the normal balance between the homeostatic and hedonic control of food intake. Mice All procedures were approved by the veterinary office of Canton de Vaud (Switzerland) under the veterinary license number VD2513. Slc2a2 loxP/loxP, Slc2a2 /loxP ;Nes Cre/+ (NG2KO), Slc2a2-cre and Slc2a2-cre;Rosa26tdtomato mice were as described previously20-22. NG2KO and Slc2a2 loxP/loxP mice were on a mixed C57BL/6;SV129 background; Slc2a2-cre and Glut2-cre;Rosa26tdtomato mice were on a C57BL/6 background. All studies used littermates as controls. For all experiments, mice were agematched and randomly assigned to experimental groups to ensure a non-biased animals distribution. No blinding was used. Unless otherwise stated, animals were collectively housed (maximum 5 individuals per cage) on a 12-hour light/dark cycle (lights on at 7 am) and fed with a standard chow (Diet 3436, Provimi Kliba AG). Viral vectors All viral constructs were produced by the Vector Core of the Gene Therapy Center at the University of North Carolina (UNC, North Carolina, USA). Operant conditioning Adult NG2KO males and control littermates (10 to 15 weeks old) were housed individually under a 12-h reversed light/dark cycle (lights on at 8:30 pm) at a constant temperature (22 ± 1°C) and had ad libitum access to water. Starting from the first day of experiment, mice were slightly food restricted and received ~3 g of laboratory chow per day. No more than 15 % of body weight reduction over the entire duration of the experiment was allowed. Mice were trained in operant conditioning chambers (Med Associates) under a fixed ratio 1 schedule of reinforcement (FR1) during 30-min daily sessions. Animals had choice between two nosepoke ports, an active nosepoke hole associated with a 3-sec light cue and a concomitant delivery of a liquid reward through a central spout equipped with infrared head entry detector and an inactive nosepoke hole that remained inoperative. Each active nosepoke triggered the delivery of 10 l of a 10 % sucrose or 0.2 % saccharin solution. Liquid rewards remained available for 3 sec once access to the central spout was detected. Supplementary entries in the active nosepoke in the absence of head entry detection above the liquid dipper and entries in the inactive nosepoke were recorded but had no further consequence. After a training period of 7 to 10 days, the animals were engaged into three consecutive sessions (one 90 min session per day) under a progressive-ratio schedule of reinforcement (PR). Under this schedule the mice were required to progressively increase the number of active nosepokes between two successive rewards based on the progression sequence given by23: response ratio (rounded to nearest integer) = (5e (reward 0.2) ) − 5. Hence, the progressive-ratio schedule followed the progression: 1, 2, 4, 6,9,12,15,20,25,32,40,50,62,77,95,118,145,178, 219, etc. The maximal number of active nosepokes performed to reach the final ratio was defined as the breakpoint, a value reflecting animals' motivation to get the reward. Active/inactive nosepokes and reward obtained were monitored online using an analog/digital interface coupling the operant chambers to a computer running the MED-PC behavioral software suite (Med Associates). Neuronal retrograde tracing Four adult Slc2a2-cre;Rosa26tdtomato males (10 weeks old) were anesthetized with isoflurane (5% (w/v) for induction followed by 2.5% for maintenance (Attane, Piramal Healthcare)) and placed in a stereotaxic frame (David Kopf Instruments). Nucleus accumbens (NAc) was infused bilaterally with cholera toxin subunit B (CTxB) conjugated with AlexaFluor 488 (Molecular Probes, Life Technologies). The following stereotaxic coordinates were used: AP +1.5 / ML ± 1.4 / DV -4,5 mm (10° angle). A volume of 0.4 l of 0.25 % CTxB solution was injected in each hemisphere through 33 Gauge stainless steel injector (Hamilton) coupled to a precision pump (Harvard Apparatus) with a injection rate of 0.1 l. min -1. Two weeks later, animals were anesthetized (isoflurane) and perfused transcardially with a 0.1 M phosphate buffer (PBS, pH 7.4) followed by 4% paraformaldehyde in PBS. The perfused brains were dissected and postfixed with 4% paraformaldehyde in PBS at 4 °C for 1 h and then placed overnight at 4 °C in a solution containing 30% sucrose in PBS. Brains were frozen in isopentane and maintained at -20 °C until further analysis. Twenty-five micrometers cryosections were prepared (Leica) and mounted onto glass slides with Mowiol anti-fade medium (Sigma-Aldrich). Images were taken using an Axio Imager D1 microscope interfaced with a Axiovision software or a LSM 510 Meta inverted confocal laser scanning microscope with LSM software 3.5 (Zeiss). For neuronal quantification, four sections containing PVT were screened in each mouse (n = 4) and Glut2 neurons were counted (n = 891) to determine which percentage of those neurons were colabeled with CTxB. Injection sites were verified for each mouse at the end of the experiment. Electrophysiology 4 to 8 weeks old males used for electrophysiology were deeply anesthetized with isoflurane prior to decapitation, brain removed and immediately submerged in a ice-cold slushy ACSF solution saturated with 95 % O 2 / 5 % CO 2 containing (in mM): 125 NaCl, 2.5 KCl, 1.25 NaH2PO4, 1 MgCl2, 2 CaCl2, 26 NaHCO3, 12.5 sucrose and 5 mM glucose (300 ± 5 mOsm). Acute coronal sections (250 m) containing PVT or NAc were obtained using a vibratome (VT1000S; Leica) and maintained at 32°C for at least 1 h before recording. Experiments were performed using an upright epifluorescence microscope (BX51WI; Olympus) mounted on a motorized stage and coupled to micromanipulators (Sutter instruments). Brain slices were placed in a recording chamber and continuously superfused at a rate of 2 ml. min -1 with oxygenated ACSF maintained at 32 -34 °C. Borosilicate pipettes (resistance = 3 to 5 M; Harvard Apparatus) were shaped with a horizontal micropipette puller (Sutter Instruments). Firing rate of PVT neurons at different extracellular glucose concentrations (5 or 0.5 mM) was monitored after a 10-15 min baseline through loose cell-attached recordings in current-clamp mode using a MultiClamp 700B amplifier (Molecular Devices). Intra-pipette solution contained (in mM): 130 potassium gluconate, 10 HEPES, 0.2 EGTA, 5 NaCl, 1 MgCl, 10 Na-phosphocreatinine, 4 MgATP and 0.5 Na 2 GTP (pH 7.2-7.3, 275 ± 5 mOsm). When mentioned in the text, a cocktail of synaptic inhibitors was used and was composed of picrotoxin (100 M; GABAA receptor antagonist), DNQX (10 M; AMPA and kainate receptor antagonist), DL-AP5 (100 M; NMDA receptor antagonist) and strychnine (1 M; glycine receptor antagonist)(Tocris Bioscience). For ex vivo optogenetic experiments in medium spiny neurons of the NAc, neurons were voltage clamped at −70 mV in whole-cell configuration and light-induced currents were recorded with a intra-pipette solution containing (in mM): 117 cesium methanesulfonate, 20 HEPES, 0.4 EGTA, 2.8 NaCl, 5 TEA-Cl, 2.5 MgATP and 0.5 Na 2 GTP (pH 7.2-7.3, 275 ± 5 mOsm). Blue light (473 nm) pulses were delivered on the brain slice through the microscope objective. TTX 1 M and DNQX 10 M were bath applied in the perfusion chamber. Currents traces were constructed by averaging 15 consecutive photocurrents. Signals were filtered at 2 kHz, digitized at 10 kHz and collected online using a pClamp 10 data acquisition system (Molecular Devices). Optogenetics 10 to 12 weeks old adult Slc2a2-cre or Slc2a2 +/+ males were used for optogenetics experiments. Under isoflurane anesthesia mice were placed in a stereotaxic frame (David Kopf Instruments) and PVT was infused with the viral vector rAAV2-Ef1-DIO-hChR2(H134R)-eYFP (titer = 3.08 x 10 12 particles. ml -1 ) to allow the expression of channelrhodopsin-2 in a Cre-dependent manner (0.25 l delivered at 0.1 l. min -1 ). To ensure a good expression of the ChR2 in the entire PVT the virus was injected at two coordinates: AP -0.4 / ML +0.8 / DV -3.5 mm and AP -1.6 / ML +0.7 / DV -3 mm with a 10° angle to avoid the damage to the superior sagittal sinus. Three weeks after viral injection mice were either used for ex vivo electrophysiological experiments or involved in in vivo operant conditioning behavioral tasks as described above. For ex vivo experiments, 4 ms light pulses were delivered by a high-power collimated blue LED system (M470L2-C1 + LEDD1B; Thorlabs) through the optical path of the microscope. For in vivo experiments, optical fiber cannulas (Precision Fiber Products) were lowered either unilaterally in PVT (AP -0.4 / ML +0.8 / DV -3.4 mm, 10° angle) or bilaterally in NAc (AP +1.5 / ML ±1.35 / DV -4.2 mm, 10° angle) and fixed to the skull with surgical screws (PlasticsOne), tissue adhesive (VetBond; 3M) and dental cement (Paladur; Heraeus-Kulzer). A DSPP laser (LRS-0473-GFM-00100-05; Laserglow Technologies) was used to deliver light pulses (10 ms light pulses at 20 Hz, 1 s on/1s off, 10-15 mW) through an optical fiber (0.22 NA, 200 m core diameter; Doric Lenses) that was attached to the optic cannula at the beginning of each experimental session (FR1 or PR). Light-stimulation was applied during the entire duration of the experimental session. To avoid the optical fiber from twisting and to allow mice to move freely, a rotary joint (FRJ 11; Doric Lenses) was placed outside the operant chamber right on top of the animal. Injection sites and optical fiber cannula placement were verified for each mouse at the end of the experiment. PVT Glut2 deletion 10 to 15 weeks old adult Slc2a2 loxP/loxP males were injected with the viral vectors rAAV5-Cre-GFP (titer = 5 x 10 12 particles. ml -1 ) or rAAV5-TRUFR-GFP (titer = 6.9 x 10 12 particles. ml -1 ) at the following injections sites: AP -0.4 / ML +0.8 / DV -3.5 mm (10° angle) and AP -1.6 / ML +0.7 / DV -3.0 mm (10° angle). Each injection consisted in an infusion of 0.25 l virus at a speed of 0.1 l. min -1. At the end of the operant conditioning experiments, mice were anesthetized (isoflurane), sacrificed and deletion of Glut2 was determined via PCR amplification of genomic DNA from dissected brain tissue containing PVT as previously described24. PCR amplification gels were made in duplicates. Injection site was verified for each mouse at the end of the experiment. Statistical analysis Statistical significance was calculated using GraphPad Prism 5.0c (GraphPad Software). The sample size for each experiment was determined based on published studies using similar experimental design together with pilot experiments from our laboratory. This allowed us to determined the sample size required for each experiment to ensure a statistical power of 0.8 and an alpha level of 0.05. Two-sided Student's t-tests were used to analyze statistical significance between two experimental groups. To ensure variance similarity between compared groups, a Fischer's F-test was performed before each t-test. When multiple comparisons was required, one-way or two-way ANOVA was performed followed by a Bonferroni's post-hoc test. The data distribution was assumed to be normal. Data are represented as means ± s.e.m. The levels of statistical significance are indicated as follow: *** P<0.001 ; ** P<0.01 ; * P<0.05. Data availability Data are available upon request from the first author (Bernard.Thorens@unil.ch). Europe PMC Funders Author Manuscripts Europe PMC Funders Author Manuscripts

The effective performance measures for store managers on restaurant chain growth Chain organization has become the most prevalent form of the restaurant industry. Facing asymmetric information, however, many chains encounter difficulties in monitoring chain store managers who serve different markets. A proper incentive scheme for store managers must be designed to solve the agency problem. However, measuring performance is a critical element of any incentive system. Thus, this study examined the appropriate performance measures of assessing the performance of a store manager. Results revealed that emphasizing quality-related performance measures of assessing store managers produces significant effects on chain growth. Furthermore, placing greater emphasis on quality-related performance measures produces a greater effect on chain growth if the operation can be easily standardized. Contrary to the negative effect of stock-based compensation, bonus-based compensation could strengthen the positive relationship between quality-related performance measures and chain growth. The alignment among quality-related performance measures, operation standardization, and reward modes was considered crucial for managing restaurant chains.

Inadequate Reference Datasets Biased toward Short Non-epitopes Confound B-cell Epitope Prediction* X-ray crystallography has shown that an antibody paratope typically binds 1522 amino acids (aa) of an epitope, of which 25 randomly distributed amino acids contribute most of the binding energy. In contrast, researchers typically choose for B-cell epitope mapping short peptide antigens in antibody binding assays. Furthermore, short 611-aa epitopes, and in particular non-epitopes, are over-represented in published B-cell epitope datasets that are commonly used for development of B-cell epitope prediction approaches from protein antigen sequences. We hypothesized that such suboptimal length peptides result in weak antibody binding and cause false-negative results. We tested the influence of peptide antigen length on antibody binding by analyzing data on more than 900 peptides used for B-cell epitope mapping of immunodominant proteins of Chlamydia spp. We demonstrate that short 712-aa peptides of B-cell epitopes bind antibodies poorly; thus, epitope mapping with short peptide antigens falsely classifies many B-cell epitopes as non-epitopes. We also show in published datasets of confirmed epitopes and non-epitopes a direct correlation between length of peptide antigens and antibody binding. Elimination of short, ≤11-aa epitope/non-epitope sequences improved datasets for evaluation of in silico B-cell epitope prediction. Achieving up to 86% accuracy, protein disorder tendency is the best indicator of B-cell epitope regions for chlamydial and published datasets. For B-cell epitope prediction, the most effective approach is plotting disorder of protein sequences with the IUPred-L scale, followed by antibody reactivity testing of 1630-aa peptides from peak regions. This strategy overcomes the well known inaccuracy of in silico B-cell epitope prediction from primary protein sequences. Knowledge of B-cell epitopes of proteins is essential in many fields of applied biomedical research, such as antibody diagnostics and therapeutics, vaccines, as well basic research. Laboratory methods for identification of such epitopes are time-consuming and labor-intensive. Hence, any reduction in the need for discovery and confirmatory wet-lab research by epitope prediction algorithms is highly desirable. Among in silico predictive methods from primary sequence information, epitope prediction algorithms are distinguished for their lack of reliability. This underperformance prompted us to examine current approaches to B-cell epitope prediction by use of extensive data on epitopes and confirmed non-epitope regions of the Chlamydia spp. proteome, accumulated in research on chlamydial molecular serology. Recent three-dimensional antibody-antigen complex studies show that about 15-22-aa 2 antigen peptide residues are structurally involved in binding of epitopes to 17-aa residues in antibody complementarity-determining regions (CDRs; paratopes). Among these 15-22 structural epitope residues, about 2-5 aa, termed functional residues, contribute most of the total binding energy to antibodies. These functional residues lie only in a very small fraction of B-cell epitopes closely spaced to each other and embedded among the structural residues, representing the classical concept of continuous B-cell epitopes. In the vast majority (90%) of B-cell epitopes, functional as well as structural residues are randomly distributed within 15-150-aa linear antigen sequences, essentially representing discontinuous epitopes. Thus, a peptide antigen can effectively bind an antibody only if it contains the majority of the functional residues, and only a small fraction of the short peptides of 4 -11 aa will contain sufficient functional residues for high affinity binding. Therefore, short peptide targets in B-cell epitope mapping and prediction may represent an inherent, unsolvable conundrum, because most of these short peptides, even from proven dominant epitope regions, will fail to bind antibodies strongly and therefore will give many falsenegative (non-epitope) results. Mammalian immune systems can be forced to generate antibodies against virtually any molecule, regardless of antigen origin, by using excessive amounts of adjuvants and antigens. However, the antibody response did evolve in response to infections that generate much lower antigen exposure, thus antibodies may be preferentially directed toward proteins and peptide regions with certain biological, structural, and physiochemical properties that determine optimal epitopes. Antibody formation during an immune response to any given epitope is inherently stochastic due to the random availability of a cognate B-cell receptor within the large pool of circulating B-cells, all with different B-cell receptors generated by recombination of the immunoglobulin gene. Another level of stochasticity in the antibody response to any given protein is the exposure of a protein to the immune system. Wang et al. report that only 4.2% of about 900 Chlamydia trachomatis (Ctr) proteins induce natural antibody responses in 40% of human hosts. Therefore, any peptide of the remaining 95.8% non-immunodominant proteins is unlikely to elicit antibodies, regardless of its B-cell epitope properties. Hence, for accurate evaluation of epitope prediction methods, epitope/non-epitope data should be derived from testing of known immunodominant proteins, with multiple rather than single sera to account for the stochasticity of the antibody response. B-cell epitope prediction has been first based on various properties of individual amino acids (aa) such as hydrophilicity, hydrophobicity, solvent accessibility, flexibility, or ␤-turn propensity, and combinations thereof (10 -16). However, even the best combinations of aa propensity scales performed only marginally better than random sequence selection. With the availability of B-cell epitope databases, antigenicity scales and machine learning approaches (19 -24) have been attempted, and improved prediction accuracy has been reported. Nevertheless, due to epitope redundancy, the predictive power may have been overestimated because these algorithms performed poorly on independent data. Therefore, B-cell epitope prediction algorithms must be evaluated on independent datasets that had not been used to train/develop the algorithms/scales. The ever increasing number of solved three-dimensional protein structures has allowed the development and testing of numerous complex algorithms for prediction of physicochemical and structural properties of proteins directly from the aa sequences. Among these properties (scales), disorder tendency describes protein regions without defined three-dimensional structure that are inherently flexible, hydrophilic, solvent accessible, and thermally mobile (high B-factor). Incidentally, all of these properties are shared with B-cell epitopes ; thus, protein disorder tendency is a prime candidate scale for B-cell epitope prediction due to its multifaceted properties. This investigation is an extension of a comprehensive study that identified immunodominant B-cell epitopes of Chlamydia spp.. After encountering numerous failures of in silico B-cell epitope prediction, we used the first principles established above to analyze the shortcomings of B-cell prediction methodology. Using pools of hyperimmune mouse sera, we determined epitope/non-epitope regions of immunodominant Chlamydia spp. proteins by use of long 16 -40-aa peptide antigens. These data created epitope/non-epitope datasets for accurate testing of numerous B-cell epitope prediction or aa/protein property algorithms/scales (henceforth termed scales). Subsequent testing revealed that public datasets were biased toward short epitope/non-epitope antigens, and removal of these short antigens dramatically increased prediction accuracy of most scales. We show that in general machine learning methods cannot predict epitopes with high accuracy; rather, many scales designed for prediction of protein properties, particularly disorder tendency, identify B-cell epitopes with better accuracy. B-cell Epitope Peptide Reactivity with Anti-chlamydial Hyperimmune Sera-Hyperimmune sera were raised in mice as described previously. Briefly, 9 -50 mice were challenged three times with high but non-lethal intranasal chlamydial inocula, to mimic antibody production following natural infections. Bovine sera used were obtained from animals with PCRconfirmed natural Chlamydia spp. infection. Peptide antigens were chemically synthesized with N-terminal biotin, captured onto streptavidin-coated microtiter plates, and incubated with hyperimmune sera. Primary antibodies were detected with horseradish peroxidase-conjugated secondary antibodies in chemiluminescent ELISA, and data were expressed as relative light units/s (rlu/s) and for ease of display were divided by 1,000 (rlu/s 10 3 ). All peptides were analyzed on white microtiter plates by use of specific positive pooled hyperimmune sera as well as negative control sera in wells coated with specific peptides and in a non-coated well. For the final background-corrected results, 150% of the background signal (mean 2 S.D.) in the non-coated well of each serum was subtracted from its specific peptide signals. To avoid false-positive results in quantitative evaluation of the reactivity of any peptide with individual mouse sera, and with bovine sera from naturally infected cattle, we used a more stringent cutoff of 10,000 rlu/s. Overall methods are described in detail by Rahman et al.. B-cell Epitope/Non-epitope Datasets-These datasets are described briefly below, and a detailed description is provided in supplemental Table S1, and sequences are provided in the supplemental Appendix. Concatenated Epitope/Non-epitope Virtual Proteins-Epitopes and non-epitopes of the Lbtope_Fixed_non_redundant and Lbtope_ Confirm datasets were grouped by sequence length. Concatenated polyproteins of the sequences of each group were constructed by randomly combining all sequences. Similarly, concatenated polyproteins of Swiss-Prot sequences were constructed by randomly combining all sequences of the BCP12, BCP14, or BCP18 datasets. Concatenated Virtual Proteins of 50-Amino Acid-extended Epitopes/Non-epitopes Embedded in Random Sequences-All 16 -20-aa epitopes of Lbtope_Confirm and fbcpred.pos.nr80 datasets were extended symmetrically to 50 aa with source protein sequences. These fragments were interspersed with random 150-aa Swiss-Prot sequences into a concatenated virtual polyprotein. Similar polyproteins were assembled from all 16 -33-aa non-epitopes of the Lbtope_Confirm dataset and all epitopes/non-epitopes of the Chl_18Prot and Chl_43Prot datasets. For assembly of additional non-epitope datasets, random 50-aa peptides of the Bcpreds epitope source proteins (Bcpreds_ Prot), Swiss-Prot proteins, and the C. trachomatis proteome were similarly linked with 150-aa interspacing sequences into concatenated polyproteins. B-cell Epitope/Non-epitope Annotation of Individual Chlamydia spp. Proteins-All residues of 18 immunodominant proteins of Chlamydia spp. were annotated in the Chl_18Prot dataset as Pos (positive, epitope), Neg (negative, non-epitope), or NT (not tested, unknown epitope status). The annotation is based on the reactivity of 16 -20-aa peptide antigens with murine and/or bovine sera. For peptide datasets, 10-aa-spaced peptides of these proteins were used. Computation of Amino Acid Residue Scores for Physicochemical, Structural, and Evolutionary Protein Properties-Websitebased freeware algorithms/scales (10 -15, 17-20, 22-24, 29 -55) for protein properties were used to calculate individual residue scores for aa sequences of individual or polyproteins. Moving window scores were assigned to the center residue of the particular window. When required, missing scores for Nand C-terminal residues were inserted using scores of the adjacent residues. If algorithms/scales did not provide an output score for internal residues/windows, the minimum score of this protein was inserted. The polymorphism score was calculated by inverting the multiple sequence conservation score of the AACon algorithm in the Jalview freeware. Comparison of Receiver Operating Characteristic (ROC) Curves of Protein Property Scales for B-cell Epitope Prediction- Bimodal epitope/non-epitope classification was achieved by F test classification based on the linear predictor variable in discriminant analysis with the software package JMP Pro 11 (SAS Institute Inc., Cary, NC). This software was also used to construct ROC curves and calculation of area under the ROC curve (AUC) for ranking of protein property scales for B-cell epitope prediction. Data formatting was performed in Microsoft Office Excel 2013, and all additional statistical analyses were performed by the Statistica 7.1 software package (Statsoft, Tulsa, OK). Differences between means of peptide reactivities and/or background were analyzed by one-tailed paired Student's t test, and p values 0.05 were considered significant. The significance of differences between B-cell epitope prediction scales was tested by one-tailed paired Student's t tests of AUCs of ROC analyses. If multiple independent test datasets were available, the mean AUC values of these scales for these datasets were compared. If multiple proteins of a single dataset were available, the mean AUC values for these proteins were compared. If ROC curves for different B-cell epitope prediction scales were analyzed for a single dataset, specificity was sampled in 0.05 increments for sensitivities from 0.05 to 0.85 and in 0.02 increments from 0.90 to 0.98, and the specificity means were compared, or the mean accuracy values at 40, 60, 80, 90, and 95% sensitivity were compared. Antibody Binding Increases with Length of Peptide Antigens- Within a peptide B-cell epitope, 15-22 residues are typically structurally involved in antibody binding. Sivalingam and Shepherd reasoned that clustering or random distribution of the structural residues would determine the length of peptide antigens required for antibody binding. In this study, we tested length-dependent peptide antigen reactivity for previously identified epitope regions of the chlamydial outer membrane protein A, OmpA. Seven-12-aa peptide antigens invariably produced lower ELISA signals than longer ones (Fig. 1A). Occasionally, extensive elongation of peptide antigens may mask structural residues and reduce the signal relative to a slightly shorter peptide antigen of optimum length ( Fig. 1A; 32versus 24-aa Cpe peptides). To quantify the effect of peptide length on antibody binding, peptide antigens of different lengths from 17 epitope regions of OmpA and inclusion membrane A (IncA) proteins of Chlamydia spp. were tested (Fig. 1B). Compared with short 7-12-aa peptides, intermediate 16-aa peptides produced 1.8-fold ELISA signal intensity, and long 24 -40-aa peptides produced 4.1-fold signal intensity (p value 10 2, one-tailed Student's t test, relative log-transformed signal). Importantly, the main 14.3-fold reactivity increase was achieved by elongation of the 13 lowest reactive short peptides from 7-12 to 24 -40 aa (p value 10 4 ), whereas elongation of the 13 lowest reactive intermediate 16-aa peptides from 24 to 40 aa produced a moderate 3.3-fold increase (p value 10 4 ). To identify optimum peptide antigen lengths, we tested the central 16-and 24 -40-aa peptide antigens of 55 unique epitopes on 28 Chlamydia spp. proteins with pooled mouse sera (Fig. 1C). As observed before, 20% of elongated peptides produced a reduced signal, presumably due to epitope masking. However, long 24 -40-aa peptides produced on average a 2.1fold higher signal than the corresponding 16-aa peptides (p value 10 4 ). The reactivity of the 28 lowest reactive 16-aa peptides increased 9.1-fold for the respective long 24 -40-aa peptides (p value 10 4 ). To confirm the host independence of length-dependent peptide reactivity, another set of chlamydial peptides yielded equivalent results with bovine sera (Fig. 1D). Evaluation of B-cell Epitope Prediction Algorithms Confounded by Over-representation of Short False-negative Epitopes in Public Datasets-Many investigators typically use short peptide antigens of 4 -11 aa for epitope mapping, with results added to public reference databases that are used for the development of B-cell epitope prediction algorithms/scales (10 -24). These datasets may therefore be biased toward short epitopes and many false-negative epitope determinations due to the marginal antibody binding of short peptides. This may explain the poor, close to random, epitope prediction accuracy that most epitope prediction scales show in practical application, even if they scored highly in evaluation with public datasets. We hypothesized that removing short epitope/nonepitope sequences from public datasets would allow correct performance ranking of B-cell epitope prediction scales. To test this hypothesis, we used the "Lbtope_Variable_non_redundant" dataset with 8,011 B-cell epitopes and 10,868 nonepitopes, retrieved by Singh et al. from experimentally validated epitopes as well as non-epitopes from the Immune Epitope Database (IEDB). Importantly, 5-10-aa non-epitopes include 50% and 5-16-aa non-epitopes include 80% of all non-epitopes deposited in the parent IEDB. Among the 6 -11-, 12-15-, 16 -20-, and 21-30-aa sequences, the Lbtope_ Variable_non_redundant dataset contains 2.12, 1.49, 0.93, and 0.54 numbers of non-epitopes compared with epitopes. These data indicate that short non-epitope sequences are over-represented in the public knowledge base. For analysis shown in Fig. 2, epitopes and non-epitopes were grouped by length, and all sequences of each length group were randomly concatenated into a single virtual protein. Hydrophilicity of all consecutive non-overlapping 20-aa peptide windows of each concatenate was predicted by use of the Parker ProtScale in ExPASy, a parameter used in B-cell epitope prediction. Results in Fig. 2A show that in the 12-15-, 16 -20-, and 21-30-aa length concatenates, the hydrophilicity scores of epitope and non-epitope virtual proteins differ highly significantly (p value 10 6, Student's t test). In contrast, the hydrophilicity of epitope and non-epitope virtual proteins is not different for the 6 -11-aa length concatenates (p value 0.052). Thus, for peptides longer than 11 aa, the hydrophilicity scale discriminates between epitopes and non-epitopes but not for shorter peptides. Similarly, with values of 0.50 for the AUC ROC curve, additional scales in Fig. 2B show random distribution of epitope versus non-epitope prediction for the 6 -11-aa concatenates. In contrast, these scales show highly significantly increased prediction accuracy for concatenates of peptides longer than 11 amino acids, indicating significant discrimination between epitopes and non-epitopes (Fig. 2B). Analysis of the "Lbtope_ Confirm" subset, composed of epitopes/non-epitopes that were at least twice independently experimentally validated, con- FIGURE 2. B-cell epitope prediction score and performance in dependence on peptide length. A, hydrophilicity scores of epitopes and nonepitopes are grouped by length in the Lbtope_Variable_non_redundant dataset. Hydrophilicity (Parker) scores were obtained by using default settings in the ProtScale tools of the ExPASy server. Length-dependent hydrophilicity (95% CI) of epitopes and non-epitopes is shown in green and red, respectively, and the p values for differences are shown in green. B, epitope length-dependent prediction performance (area under receiver operating characteristic curve) of different prediction scales in the Lbtope_Variable_non_redundant dataset. ***, p value 10 6 for comparison of any scale to any other scale of 6 -11-aa epitopes versus longer epitopes. Hydrophobicity (Miyazawa,30), a ProtScale for hydrophobicity; Bepipred, a hidden Markov model combined with the Parker hydrophilicity scale ; IUPred-L, an algorithm for protein disorder tendency. C, all 6 -20-aa epitopes and non-epitopes of the Lbtope_Confirm dataset grouped into 6 -11-, 12-15-, and 16 -20-aa peptides are compared with Swiss-Prot 12-, 14-, and 18-aa peptides of the Bcpreds BCP12, BCP14, and BCP18 datasets. The p value for hydrophilicity score differences between epitopes and non-epitopes is shown in green and between non-epitopes and Swiss-Prot random peptides in blue. All epitopes have higher hydrophilicity scores than Swiss-Prot random peptides (p value 10 5 ). firmed the result of the Lbtope_Variable_non_redundant dataset ( Fig. 2C). An additional finding is that the 16 -20-aa nonepitopes do not have a significantly higher score than the random Swiss-Prot peptides, although the 6 -11-and 12-15-aa non-epitopes do (Fig. 2C). This suggests that the Lbtope_Confirm dataset may have a high frequency of incorrect identification of short 6 -15-aa peptides, particularly 6 -11-aa, as non-epitopes. Evaluation Accuracy for B-cell Epitope Prediction Depends on Epitope/Non-epitope Discrimination in Test Datasets-Ideal algorithms/scales for prediction of B-cell epitopes should discriminate known epitopes from experimentally validated nonepitopes and identify epitopes within complete source proteins and proteomes. Since prediction accuracy should ideally be validated with multiple datasets, we evaluated the prediction performance by use of four positive datasets of experimentally validated B-cell epitopes and five negative datasets of experimentally validated non-epitopes or of random peptides from proteomes ( Table 1). All 16 -20-aa epitope/non-epitope sequences of the datasets were centered within their 50-aa source protein sequences, and these 50-aa sequences were randomly concatenated into a single virtual protein, interspersed with random 150-aa sequences from the Swiss-Prot database (supplemental Table S1 and Appendix). For evaluation of B-cell epitope prediction, we used the original score of each algorithm with default settings for each amino acid residue, thus each epitope/non-epitope sequence received individual scores for the 20 central residues. Correct or incorrect epitope prediction of all residues was evaluated by AUC in ROC analysis. In Table 1, the column for each scale indicates epitope versus non-epitope discrimination (AUC in ROC analysis) of the scale for each compared combination of positive and negative datasets. The average AUC column, next to the rightmost column, indicates epitope/non-epitope discrimination averaged over all tested scales and therefore ranks the combined discrimination in both positive and negative datasets. The four positive datasets can be ranked by their AUC in comparison with the negative Swiss-Prot or Ctr-Proteome datasets, clearly showing significantly higher discrimination for both chlamydial datasets than for the fBcpreds and Lbtope_Confirm datasets (p value 0.02, one-tailed paired Student's t test). The average AUC row, next to the bottom row in Table 1, indicates epitope/non-epitope discrimination averaged over the 12 tested pairs of datasets and shows that disorder tendency -cell epitope prediction accuracy (AUC of ROC curves) in dependence on the evaluation dataset a Datasets with experimentally identified epitopes/non-epitopes or random peptides are shown (see supplemental Table S1). b AUC data are shown for the best performing scale of each category as defined for all scales tested (see supplemental Tables S2 and S3). c Antigenicity scale, IEDB tool for antibody epitope prediction. d Support vector machine model (SVM) trained on the Lbtope_Confirm dataset. e Surface accessibility scale, IEDB tool for epitope prediction. f ␤-turn scale, IEDB tool for epitope prediction. g Average of seven propensity scales for epitope prediction. h Flexibility scale, IEDB tool for epitope prediction. i Quality of scales or datasets was ranked by AUC, with rank number determined by 1 for the highest AUC and addition of 1 for each 0.01 AUC reduction; antigenicity and Lbtope scales were excluded from ranking because antigenicity is a negative predictor and Lbtope was trained on the analysis dataset. j Not applicable for quality ranking because the datasets served to train the Lbtope support vector machine model. k Highest AUC in the compared dataset. discriminated best of all algorithms tested (AUC 0.75 for IUPred-L ), highly significantly better than Bepipred (AUC 0.70), the next-best algorithm in the 12 AUC comparisons of positive and negative datasets (p value 10 3, onetailed paired Student's t test). Since the machine learning Lbtope algorithm was trained on the Lbtope_Confirm dataset, it performed extremely well in this dataset (AUC 0.97, 0.84, and 0.81), but very poorly, close to randomization, in all other datasets (average AUC 0.57). In contrast, the protein disorder scale IUPred-L consistently discriminated best (Table 1), with AUCs depending on the datasets (0.58 -0.88). Wide Amino Acid Context and Standardized Scoring Maximize B-cell Epitope Prediction Accuracy-Most amino acid propensity and B-cell prediction scales score the context-dependent epitope likelihood for each amino acid residue by averaging the adjacent 4 residues (10 -19). In contrast, protein disorder prediction operates in a wider sequence context. To simulate the wider sequence context, we asked the question if scores for long peptides improved prediction accuracy of narrow-context scales and, if so, what the peptide length dependence of such an improvement was. In Table 2, we determined B-cell epitope prediction accuracy for single scores for the central 1-aa epitope/non-epitope residue and contrasted it to prediction by single average scores for the central 10-, 15-, 20-, 25-, or 30-aa residues. The mean AUC values of all 12 comparisons of four to five non-epitope datasets (as in Table 1) are shown in Table 2. The results indicate optimal B-cell epitope prediction for 20 -30-aa peptide scores. Long peptide scoring improves prediction substantially for narrow-context scales such as hydrophilicity, hydrophobicity, flexibility, or Bepipred but not for wide-context protein disorder tendency scales such as IUPRed-L and VSL2B. Table 2 shows prediction of epitopes/non-epitopes embedded in virtual concatenated polyproteins. In this approach, dif-ferences between the widely divergent average scores of source proteins cannot be offset by standardization (i.e. set to mean 0 and S.D. 1). To eliminate B-cell prediction bias induced by selection of epitope source proteins, we analyzed scores standardized for each protein of the 18 chlamydial protein datasets (Chl_18Prot; supplemental Table S1). Table 3 shows the results for comparison of experimentally validated epitopes of these proteins to experimentally validated non-epitopes (Pos versus Neg) or the total remaining protein (Pos versus Neg NT). Standardization substantially improves B-cell epitope prediction accuracy for disorder and solvent accessibility scales, but not for individual amino acid propensity scales such as hydrophilicity or hydrophobicity. Similar to results for concatenated polyproteins (Table 2), standardized scores of long peptides improve performance of narrow-context but not of wide-context scales ( Table 3). The polymorphism scale for the 18 chlamydial protein dataset was derived by inverting the Jalview AACon conservation score calculated from multiple sequence alignments of these proteins with the available homologous Chlamydia sequences. Thus, it is completely independent of individual amino acid properties and quantifies only evolutionary sequence change at each residue. Standardization of polymorphism scores in Table 3 improves prediction because chlamydial proteins have widely divergent rates of evolution. Importantly, averaging over 25-aa residues again provides maximum prediction accuracy, suggesting that wide-context properties in general are the best predictors of B-cell epitopes from their primary amino acid sequences. Protein Disorder Most Accurately Predicts B-cell Epitopes-For epitope/non-epitope discrimination in the ROC curve in Fig. 3A, sensitivity at given specificity (or specificity at given sensitivity) of the IUPred-L disorder or the combined scale is higher than that of Bepipred or LBTope (p value from the maximum (bold red font) are shown in red font. b A single score of the central residue was considered for each peptide in the datasets shown in Table 1. c Scores of the central 10/15/20/25/30aa residues were averaged to a single peptide score. d Comparison of 1-aa (single residue) versus 10-aa peptide scoring. e Comparison of 10-aa versus 25-aa peptide scoring. F/f AUC significantly higher for 10-aa peptide scoring than for 1-aa scoring ( F, 10 6 p value 10 3 ; 10 3 p value 10 2 ; one tailed paired Student's t-test with 12 AUC values). G/g AUC significantly higher for 25-aa peptide scoring than for 10-aa peptide scoring ( G, 10 4 p value 10 3 ; 10 -3 P-value 0.023). 10 4, one-tailed paired Student's t test). Similarly, when epitopes were discriminated from the complete remaining proteins in Fig. 3B, IUPred-L scale performed significantly better than Bepipred or LBTope. In final testing of the overall prediction approach applied to the 18 individual proteins of the Chl_18Prot dataset, IUPred-L scale also best discriminated individual epitope residues from the whole remaining protein (average AUC of IUPred-L 0.91, minimum 0.74, maximum 1.00, S.D. 0.08; Table 4). Tables 1 and 2, most epitope/non-epitope sequences were derived from public datasets of variable and largely unknown discrimination accuracy. For maximum accuracy, we therefore selected the 18 chlamydial protein datasets (Chl_18Prot; supplemental Table S1) with extensively validated epitopes as well as non-epitopes on each protein, all identified in a single investigation. For the Chl_18Prot dataset, 151 standardized primary scales for B-cell epitope prediction were evaluated (supplemental Tables S2 and S3). To improve B-cell epitope prediction, investigators frequently combine scales. To test this concept, we evaluated 126 combined scales that were derived by linear combination of 2-14 standardized primary scales ( Fig. 4 and Tables S2 and S3). In Fig. 4, we asked whether the combined scales, derived from 25-aa moving averages of the primary scales, improve B-cell epitope prediction. Results show that the combination of scales only incrementally improves B-cell epitope prediction (Fig. 4). The best combination of the primary scales provides only a 3.8% improvement of prediction accuracy over IUPred-L protein disorder in five tests at 40, 60, 80, 90, and 95% sensitivities (Fig. 4C, p value 0.049, paired Student's t test, with five accuracy values). Collectively, the dominant conclusion is that the main improvement for B-cell epitope prediction References 11,19,24,30,31, cited in the table. a The Chl-18Prot dataset was analyzed. Pos, Positive (epitopes); Neg, negative (non-epitope); NT, not tested (epitope or non-epitope status is unknown). Pos versus Neg indicates epitopes were compared to non-epitopes; and Pos vs NegNT indicates epitopes were compared to the total remaining protein. Standardization of individual protein scores improves B-cell epitope prediction Average AUC values that differ by 0.01 from the maximum (bold red font) are shown in red font. b Solvent accessibility (ASA_Spine-X) residue solvent accessibility ; polymorphism is sequence divergence in multiple sequence alignment, calculated by inverting the conservation score of AACon in the Jalview freeware. c Original non-standardized score for central 1-aa residue in the peptides. These scores were obtained with individual protein sequences as input. d Original scores were standardized (mean 0 and S.D. 1) for each of the 18 chlamydial proteins, and the standardized score for the central 1-aa residue in the peptides is shown. e Difference in AUC values between standardized and non-standardized scores. f Sensitivity at a given specificity is significantly higher in ROC curves for standardized versus non-standardized scores ( f, 10 6 p value 0.01; one-tailed paired Student's t test). g Peptide scores were calculated using the average of standardized scores for the central 5-, 9-, 17-, 25-, 33-, 41-or 49-aa residues. h Difference in AUC values between standardized scores of 25-and 9-aa peptides. i Sensitivity at given specificity is significantly higher in ROC curves for 25-aa versus 9-aa standardized scores ( i, 10 -6 p value 0.01; one-tailed paired Student's t test). JULY 8, 2016 VOLUME 291 NUMBER 28 JOURNAL OF BIOLOGICAL CHEMISTRY 14591 comes from using the optimal IUPred-L primary scale (Tables 3, supplemental Tables S2 and S3, and Fig. 4). Underperformance of Machine-learning B-cell Prediction Algorithms-In evaluation of B-cell epitope prediction algorithms, scores of most physicochemical, structural, and evolutionary protein properties are higher than those of machine learning algorithms (Fig. 5). In addition, the discrimination power of all scales is higher when epitopes are tested against the remaining protein than against experimentally validated nonepitopes (Table 3 and Fig. 5A). As a consequence, the prediction performance against the remaining total protein sequences is also consistently higher for all scales. An explanation for this counterintuitive observation is that non-epitopes had initially been selected as candidate epitopes by high scores in prediction scales ( Fig. 2) but failed to react with antibodies. The higher scores for tested non-epitopes thus induced a pre-selection bias that makes evaluation of B-cell epitope prediction scales more difficult. Fig. 5B compares B-cell epitope prediction scales that were among the best combinations of the primary scales in our study with several publicly available algorithms/scales that almost uniformly perform poorly. This poor discriminatory power of machine learning algorithms most likely results from suboptimal training datasets with an over-representation of short nonepitopes. For example, Lbtope was trained on 80% short 6 -16-aa confirmed non-epitopes. In contrast, Bcpreds was trained by use of random Swiss-Prot peptides as non-epitopes, equal in length to confirmed epitopes, and they performed better than Lbtope (0.06 -0.10 AUC value difference between Bcpreds and Lbtope; Fig. 5B). Among the published combined B-cell epitope prediction scales, only Bepipred showed acceptable performance, better than the accurate Parker hydrophilicity scale (0.06 -0.09 ⌬AUC compared with Parker hydrophilicity; see Table 3). Bepipred nevertheless requires long peptide scores for optimal performance (0.07-0.10 ⌬AUC between 25-aa peptide and default scoring; Table 3), and it is not a pure machine learning algorithm because it combines a protein property scale, Parker hydrophilicity with a hidden Markov model. Dominant Properties of B-cell Epitope Regions-Our evaluation of the discriminatory power of B-cell prediction algorithms in the extensively experimentally confirmed Chl_18Prot dataset allowed us to deduce some critical global properties that define natural B-cell epitope regions. Clearly, the dominant property is the propensity for a disordered state of amino acids in B-cell epitopes. This property is linearly correlated to hydrophilicity (inverted Miyazawa hydrophobicity scale ; R 2 0.66, p value 10 6 ; linear regression analysis of the 25-aa peptide scores centered around each residue of the Chl_18Prot dataset), flexibility (Karplus and Schulz ; R 2 0.56, p value 10 6 ; solvent accessibility (Spine-X ; R 2 0.49, p value 10 6 ), evolutionary mutation rate (R 2 0.43, p value 10 6 ), coils in secondary structure (PSIPRED ; R 2 0.42, p value 10 6 ), and ␤-turns (Levitt ; R 2 0.40, p value 10 6 ). Thus, due to multi-collinearity, the multifaceted properties of protein disorder tendency synthesizes all of these properties into a single descriptor (Fig. 5). The physicochemical, structural, and evolutionary properties of B-cell epitope regions discriminate them sufficiently to translate into significant differences in amino acid composition to the remaining total proteins. B-cell epitopes are enriched for proline, followed by glutamic and aspartic acids, asparagine, threonine, alanine, and serine (Fig. 5C). Epitopes are also relatively depleted of leucine, isoleucine, tryptophan, phenylalanine, tyrosine, and cysteine. Proposed B-cell Epitope Prediction-As a result of the preceding analyses, an easily implemented approach for accurate B-cell epitope prediction has emerged that should be useful for investigators in many fields of antibody research. Fig. 6 demonstrates the application of the previous findings for B-cell epitope prediction in an actual example for which we generated epitope scanning data of the complete chlamydial protein IncA. In Fig. 6A, the default IUPred-L and VSL2B disorder scores of Table 3). Plots of epitope-positive rate versus false-positive rate for the 18 chlamydial protein dataset are shown. A, prediction of epitopes from confirmed non-epitopes (25-aa epitopes/non-epitopes spaced 10 aa). The combined scale represents the arithmetic mean of two disorder scales, IUPred-L and VSL2B, and one solvent accessibility scale, Accessible Surface Area, Spine-X. B, prediction of epitopes from the total remaining proteins (non-epitope plus non-tested regions). In both datasets (A and B), the combined scale and the single disorder (IUPred-L) scale performed best (highest sensitivity at given specificity or vice versa), significantly better than Bepipred or LBTope (one-tailed paired Student's t test, p value 10 4 ). TABLE 4 Epitope prediction accuracy (AUC) averaged for individual proteins of the 18-chlamydial protein dataset a a Original scores obtained with default options for the algorithm/scale were smoothed by a sliding window method in which the score for each residue was averaged for the adjacent 12 residues (25-aa moving window). Smoothed scores of residues were standardized for each of the 18 chlamydial proteins and discrimination of epitope residues from remaining total residues was tested for each of the 18 proteins individually. b Coils (Spine-X) indicate the coils predicted in secondary structure. the IncA protein are plotted against IncA residue number. Solvent accessibility and hydrophilicity are shown in Fig. 6, B and C. In Fig. 6D, noise was reduced by smoothing the scores as 25-aa moving averages, and comparison was improved by standardizing the data. Comparison of these IncA epitope prediction plots with actual IncA peptide reactivity in Fig. 6E clearly shows that IUPred-L predicted scores best match experimental observations and confirm the superior B-cell epitope discriminatory power of protein disorder tendency as calculated by IUPred-L. Fig. 6E displays optimal prediction approaches by the combined scores of scales shown in Fig. 6D, and smoothed and original default IUPred-L scores. While combined scores have marginally better discriminatory power (Fig. 5), for practical purposes we consider the accuracy of IUPred-L sufficient. Also, given the wide-context nature of protein disorder scales, scores are sufficiently stable to even render smoothing unnecessary, allowing direct use of default plots obtained from the IUPred-L webserver for B-cell epitope prediction. Therefore, 16 -30-aa peptide antigens for laboratory testing can be selected directly from peak disorder regions of the IUPred-L plot. Discussion The results of this study suggest strategies for B-cell epitope identification that deviate from current approaches that many investigators use. Our approach initially identifies protein regions that harbor B-cell epitopes rather than immediately focusing on identifying peptide antigens of specified length. B-cell epitope regions can be predicted with high accuracy simply by selection of the peak regions from the IUPred-L disorder plot of a protein antigen (Fig. 6E). Next, these high probability epitope regions should be confirmed with 16 -30-aa-long peptide antigens using pooled antisera. Fine mapping of highly reactive regions with overlapping 16-aa peptides, using the individually reactive antisera of the pool, identifies regions with several functional aa residues embedded among structural epitope residues. Further reduction in peptide antigen length entails mapping with very short 6 -12-aa peptides. Success at this stage relies on stochastic identification (Fig. 2) of closely spaced randomly distributed functional residues that maximally contribute to antibody binding. Antibody binding of such short peptides is, however, typically low (Fig. 1), most likely because antigens of less than 16 aa will not bind to the complete CDR of an antibody. This approach is derived from the conclusive evidence that short 7-12-aa peptide antigens of confirmed Chlamydia spp. epitopes bind antibodies poorly (Fig. 1), and therefore many of these epitopes would be falsely classified as non-epitopes if they were identified by short peptide mapping. The poor reactivity of short peptide antigens combined with data in Fig. 2 strongly suggest that many of the short non-epitopes in public B-cell epitope datasets are likely to be actual epitopes. Most investigators who develop B-cell epitope prediction algorithms/scales draw training and test datasets from public databases such as IEDB. These reference datasets are suboptimal due to over- untrained protein property scales, such as protein disorder tendency, that were developed for different reasons nevertheless predict B-cell epitopes with higher accuracy than specifically developed B-cell epitope prediction scales/machine learning algorithms (supplemental Tables S2 and S3 and Fig. 5B). A fundamental conundrum in B-cell epitope prediction is the conceptual and methodological approach that leads to the eventual identification of a B-cell epitope. Vastly preferable is the use of x-ray crystallography-solved three-dimensional structures of antigen-antibody complexes. Such data define precisely the actual determinants of a protein antigen that specifically contact an antibody, in essence the set of protein resi-dues that are buried under a cognate antibody in the antibodyantigen complex. However, only 26 -107 non-identical three-dimensional structures of antigen-antibody complexes have been generated by different investigators from the Protein Data Bank crystallographic database (3-7, 56 -62). Such data were used for training and development of several B-cell epitope prediction methods such as CEP, DiscoTope, Rapberger's method, Ellipro, PEPITO, and Epitopia (56 -62). The major shortcoming is the requirement for the three-dimensional structure of the protein antigen. In practice, this limitation is currently insurmountable because we do not know the threedimensional structure of most proteins. Datasets Confound B-cell Epitope Prediction In practice, B-cell epitopes are commonly determined by use of peptide antigens and their ability to capture antibodies. This approach does not identify which residues of the peptide are in binding contact with antibody CDR residues and which actually contribute to the antigen-antibody complex formation. Nonetheless, antibody-reactive peptide sequences, particularly those identified by systematic mapping with overlapping peptides, are commonly referred to as B-cell epitopes (16,. This terminology is justified because even non-binding residues are specifically required to provide the structural context for binding residues, thus the linear peptide sequence is still an indispensable, if not complete, characterization of a B-cell epitope. It is important, however, to understand that epitope prediction from linear peptide sequences will weigh the total combined contributions of binding (functional) and spacer (structural) amino acids to an epitope. Nevertheless, B-cell epitope prediction from the primary amino acid sequence of a protein is a valid and, for practical purposes, highly desirable approach. In addition, tens of thousands of B-cell epitope/non-epitope sequences have been deposited in IEDB. Thus, the sequence-based B-cell epitope datasets provide a viable basis for training and development of B-cell epitope prediction algorithms (10 -22). The profound conundrum for epitope prediction by use of linear peptide sequence-based methods, however, is the fact that more than 90% of all B-cell epitopes are not linear, composed of immediately neighboring binding residues, but they are discontinuous. In almost all B-cell epitopes, the typical 2-5 dominant binding residues will be discontinuously arranged randomly in the linear epitope sequence. Nevertheless, in the majority of epitopes these binding residues are still closely spaced. For instance, Sivalingam and Shepherd show that 30-aa peptides will encompass the functional residues of 75% of all B-cell epitopes. Thus, increased lengths of peptide antigens will increase the probability of capturing more of the residues of any epitope that are required for high affinity antibody binding (Fig. 1). In addition, long peptides may increase the probability of capturing different antibody clones that may bind the same epitope region differently. For instance, C. trachomatis OmpA serovar-specific peptide serology has used 6 -10-aa peptides, with inconsistent results (66 -70). In our study, we observed strong but completely serovar-specific antibody reactivity by use of 16-aa peptide antigens. Importantly, inclusion of conserved adjacent residues shared among chlamydial species, in addition to the 7-10 central polymorphic serovar-determinant OmpA residues, was required for strong, yet specific, antibody binding. Conceptually, a peptide antigen captures antibodies if it can fold to complement the binding region of the cognate antibody. Because of such structural constraints, the length of peptide antigens may also negatively influence antibody binding. For instance, if the few randomly spaced dominant binding residues are obstructed by structural constraints such as misfolding, masking by non-epitope residues, or peptide aggregation, antibody binding may be compromised. Our study clearly shows that moderate elongation of peptide antigens strongly enhances antibody binding, while more extensive elongation reduces antibody binding again in 20% of B-cell epitopes, pre-sumably by masking epitope residues (Fig. 1). The implication of this fact is that an optimal sequence length exists that most reliably discriminates between true epitopes and non-epitopes and that sequences of that length should be used to generate datasets for the development of B-cell prediction methods. A protein surface can be thought of as a continuous landscape of epitopic regions, and any region of this landscape may be identified as an epitope under specific conditions (56,. For instance, Singh et al. reported that all nonepitopes in the LBTope_Confirm dataset have been reported as "non-epitopes" in at least two studies. Yet 8.3% of these nonepitopes are reported as "epitopes" in the fBcpreds dataset. Thus, binary classification of antigen regions into epitopes or non-epitopes is problematic because all epitopes of most antigens are not known, and defining B-cell epitopes and nonepitopes is a challenging task due to the variability in epitope discovery assays and the stochastic antibody responses to protein antigens and their epitopes. Muller et al. found almost the entire histone 2A protein antigenic when they forced highest B-cell stimulation and antibody reactivity by excessive use of adjuvants and high antigen doses. In contrast, raising antisera in our study by experimental infection rather than by forced immunization very likely resulted in much lower adjuvantation and lower antigenic stimulus by physiologically processed native protein antigens. Thus, antibodies likely were generated mainly against exposed antibody-binding regions of highly expressed proteins. In addition, targeting known immunodominant proteins by the use of antisera pooled from multiple individuals maximized correct epitope/nonepitope discrimination by offsetting the inherent stochasticity of antibody formation in individuals and by minimizing falsenegative results. We observed a clear trend that certain protein regions are a more frequent source of B-cell epitopes than others, and we think that our study identified the distinctive properties of such preferentially antibody-recognized regions. Kringelum et al. determined by x-ray crystallography that hydrophobic amino acids of epitopes located closest to the antibody, and charged amino acids most distant, but that the amino acid composition of equally surface-exposed non-epitopes did not differ significantly from epitopes. However, the amino acid composition of epitopes deviated significantly from the whole protein. We compared properties of B-cell epitope regions with experimentally confirmed non-epitope regions or the remaining protein regions, but we do not know about surface exposure. Similarly, we report that many protein properties of B-cell epitope regions differ substantially from the total remaining proteins (Fig. 5), making these properties candidates for B-cell epitope prediction. Accessibility of the antigen by cognate B-cell receptors or antibodies is the central concept in molecular recognition of epitope by the paratope, and thus highly surface-exposed hydrophilic/charged epitope residues will first interact with the antibody (Fig. 5C). Although hydrophobic amino acids except for alanine, the smallest one, are under-represented in epitopes, those that are present may "provide the glue" in the final stabilization of the antigen-antibody complex by hydrophobic interaction. All non-covalent antigenantibody interactions are thought to be driven by shape complementarities in the complex formation. Thus, paratopes may interact preferentially with flexible regions of an antigen rather than with highly structured regions. Precisely because of relaxed structural constraints, such protein regions should accommodate higher amino acid substitution rates, favoring under immunoselective pressure the emergence of escape mutants. Important antigenic regions of viral and bacterial proteins have been identified as disordered regions of these protein antigens. However, x-ray crystallography studies have not specifically reported the localization of B-cell epitopes in disordered protein regions (3-7, 56 -65). The most likely explanation for this discrepancy to our results is the fact that the Protein Data Bank database is biased toward proteins of common interest that are easy to produce and crystallize. Many expressed proteins cannot be crystallized, and among the main factors for this failure is the presence of even small numbers (1-10 aa) of disordered residues that are well known to have deleterious effects on crystallization. For convenient determination of three-dimensional structures, disordered protein regions are removed from expressed proteins. As a result, disordered proteins or protein regions are rare in the Protein Data Bank database compared with whole proteomes (79 -81). Moreover, crystal packing is thought to enforce certain disordered regions to become ordered, resulting in incorrect characterization of disordered protein residues. In addition, disordered segments crystallized together with binding antibodies are usually classified as ordered structure in the antigen-antibody complex, despite their lack of ordered structure in the unbound state. Thus, datasets generated by crystallography may inherently under-represent B-cell epitopes with high disorder tendency. Protein disorder tendency has also not been proposed for B-cell epitope prediction from primary amino acid sequences, although many protein property scales, particularly aa propensity scales, have been tested and recommended for B-cell epitope prediction (16,. In our study, the IUPred-L dis-order scale has the highest epitope discriminatory power in all datasets. We explain this discrepancy by the typical experimental approach with which investigators test sequence-based epitope prediction methods as follows: wide-context disorder properties of proteins will not be correctly determined by solely analyzing the typically short peptide sequences of databases. To achieve correct results, we elongated test peptides with source protein sequences and embedded them in a wider context of random Swiss-Prot sequences (supplemental Table S1 and supplemental Appendix). As a norm in investigations addressing protein disorder, protein residues are binary-classified as either "ordered" or "disordered." In contrast, disorder prediction algorithms quantify the probability of protein disorder, and binary classification converts the prediction scores by using an arbitrary cutoff at a predetermined threshold. By these criteria, many epitopes would not classify as disordered. However, relative to the moving average score of the whole source protein, B-cell epitopes consistently score highest for protein disorder tendency. For actual B-cell epitope prediction, the IUPred-L protein disorder scale consistently performs best (87% specificity at 80% sensitivity, 86% accuracy; Fig. 4). However, if a 25-aa moving average score is used, several other protein property scales such as hydrophilicity (Parker), hydrophobicity (Miyazawa), solvent accessibility (Spine-X), or Bepipred perform similarly. In fact, scoring by narrow-context scales for long 20 -30-aa peptides reflects protein disorder tendency such as the Globplot-2 algorithm predicts protein disorder tendency by a wide-context hydrophilicity score. It is noteworthy that even the best combination of top performing scales does not substantially increase prediction performance (Fig. 4), due to multi-collinearity of these scales. The best performing combined scale (Figs. 4 and 5 and supplemental Tables S2 and S3), derived from smoothed and standardized 25-aa peptide scores of three primary scales, improves prediction accuracy only marginally (90 -92% specificity at 80% sensitivity, 88 -90% accuracy; Fig. 4). Our data show that wide-context disorder scores or long 20 -30-aa peptide scores of narrow-context scales are optimal for B-cell epitope prediction (Tables 2 and 3), consistent with the higher antibody binding of 16 -30-aa peptide antigens (Fig. 1). Compared with highly structured protein regions, disordered regions may have several functional advantages for efficient interactions with partner molecules, such as the capacity of initiating binding by long range electrostatic interactions, high flexibility, binding plasticity and speed, minimal steric restrictions in binding, and the ability to form very stable intertwined complexes (26,27,. Hence, our investigation merges theoretical advances in protein biophysics with very practical aspects of protein interaction, the identification of peptide sequences best suited for recognition by CDRs of antibodies. Author Contributions-K. S. R. and B. K. planned the experiments; K. S. R. and E. U. C. performed the experiments; K. S. R. and B. K. analyzed the data; K. S. R., B. K., and K. S. contributed reagents and essential material; and K. S. R. and B. K. wrote the paper.

Economic Security of Economic Entities in Crises The study examines a set of measures that affect the level of economic security of the country as a whole, individual regions and companys. Current economic conditions determine the state of the economy, which is determined from a position of macroeconomic control of the state over the use of national resources to protect the economic interests of the country at national and international levels, from the perspective of microeconomics resource utilisation to ensure stable operation of the company. The purpose of the study is to identify the features of the functioning of economic entities in current economic conditions of management and substantiate ways to improve their economic security. In the course of the study, methods of comparison, induction, deduction, observation, and descriptive were used. The results of the study were the substantiation of the synthesis of factors of external and internal threats to the economic security of economic entity; determination of the role of the information field and its organisation in the accumulation of relevant information to take measures to ensure a stable financial condition and long-term development, as well as a systematisation of measures of state support for business in the context of the 2020 corona-crisis. INTRODUCTION In modern economical literature, the problems associated with ensuring the economic security of an individual economic entity are not thoroughly studied. At the same time, it should be noted that the economic security of an economic entity is primarily predetermined by the existing economic security of a particular region and the state as a whole. The stable economic situation within the region and the country as a whole and the practical absence of external threats can positively influence the economic security of an individual economic entity. Consequently, the whole set of measures that affect the economic security level of the country in general, individual regions and companies are closely inter-related. In this regard, the terminology and indicators of economic security should be uniform and logically linked at all levels: federal, regional and economic entity level. DISCUSSION In the economic literature, the authors in determining the economic security of the company emphasised protection from external and internal threats, i.e. the ability of the organisation to ensure its stability, independence, sustainability and effective satisfaction of needs. L.I. Abalkin, E.M. Buchwald hold this point of view.. Other authors, including S.Yu. Glazyeva, E.A. Oleynikova, define economic security in terms of protecting their vital interests.. According to M.M. Kalashnikov, L.P. Goncharenko and others, the economic security of a company is the state of the most effective use of corporate resources to prevent threats and ensure the stable functioning of the company at present and in the future. At the same time, all authors agree that at present, each company should work to ensure economic security to protect the interests of the company from both external and internal threats in its sustainable development. Jang Yong defines the country's security as the preservation of independence, ensured by the national development and the growth of population standards. C. Murdoch clarifies that changes in income and wealth accompany the threat to security; it is reflected in the level of employment, inflation, market access, etc., which ultimately affects economic sovereignty. In cases of significant changes in external conditions to the negative side in the context of natural disasters, war, pandemics and other force majeure circumstances, the company finds itself in a difficult economic situation, to protect itself from which it is practically impossible without the support from outside the state. According to BBC NEWS Russian service, since the beginning of the pandemic, the authorities of different states have allocated huge sums of money to fight it and support various sectors of the economy (figure 1). The internal business conditions of the company, related to the organisational structure, management, information security, finances, personnel, directly depend on the company, are entirely determined and controlled by it. However, to ensure the regular and effective functioning of the company, it is necessary to monitor the external environment and interact with it. To develop a system for ensuring the economic security of a company, necessary to determine the position that the company adheres to overcoming threats or ensuring security. The choice of position determines the further direction of actions for the formation of economic security. In the first case, all attention is focused on the threat determining, its danger to the company, developing actions to protect against it. This is a condition for protecting the whole company and a separate element, the structure of the company. In the second case, attention is focused on the facts of the financial and economic activities of the company to ensure normal functioning and reduce the likelihood of not achieving the goal of its activities. Thus, this is a sufficient condition for the safety of the company to achieve its goal of the activity. Within the framework of the company's activities, a successful combination of these approaches is possible. At the same time, it should be remembered that each of them requires its methods of achieving economic security. Achieving a high level of economic security of an economic entity is impossible without a stable state of its solvency. An organisation that is not able to fully meet its obligations becomes mostly dependent. That is why the activities carried out in the organisation to achieve a high level of solvency occupy an important place in ensuring and maintaining the proper level of economic security. Crisis periods of economic development represent the greatest danger of the destructive effect of several factors on the production, technological and human resources potential of the organisation. Such factors include a poor regulatory framework, shortage of qualified personnel, significant wear of production equipment, shortage of working capital, tight monetary and fiscal policies of the state, insolvency of partners, and several others. According to its content, several types of threats to the economic security of an economic entity should be noted: catastrophic, informational, competitive and organisational. Thus, based on the correspondence of actual and normative values of technical and economic parameters and magnitude of deviations from the threshold values of economic security organisation may be: normal, when the indicators of economic security are within thresholds; pre-crisis, when the values of at least one of the indicators of economic security threshold, while others approached the barrier values crisis, when there are excess barrier values most significant indicators of economic security; critical when violated almost all the threshold values of economic safety of an economic entity. The main functional components of the economic security of the organisation can be represented by financial, political and legal, technical and technological, information, environmental and power. At the same time, the financial component is characterised by the rational use of fixed and working capital, the capital of the organisation, the optimal exchange rate of securities on the stock exchange. The technical and technological component is characterised by the compliance of the technologies used in the organisation with international standards, their competitiveness and the possibility of further improvement. The political and legal component is related to the compliance of both the organisation and its employees with the necessary legal requirements. In the modern world, the information component of the economic security of an economic entity is critical. At the same time, information and analytical support of the organisation's activities require specific operations related to the prediction of potential threats, the collection of official information, its elaboration and sorting, as well as measures to prevent unauthorised access to information. Recently, the requirements for the environmental component have been tightened in terms of cost optimisation for compliance with environmental standards for technologies and products, minimising the cost of fines for violating environmental legislation. The creation of a system for monitoring the state and dynamics of development of organisations contributes to the early warning of danger and the adoption of necessary measures for protection and counteraction. Monitoring Advances in Social Science, Education and Humanities Research, volume 527 should be the result of the interaction of all interested services of the company. When monitoring must be a principle of continuous monitoring of the condition of the monitored object based on the current condition and trends in the development of its potential, and General economic development, the political situation and the actions of other system-wide factors. RESULTS The economic realities of the existence of the country's economy since 2014 in the context of severe economic sanctions have led to a new approach to the definition of economic security. In the current crisis conditions, economic entities had to not only continue their activities, providing various sectors of the economy with the necessary, but also quickly rebuild, be able to resist the current situation quickly. A new regulatory act was developed and adopted -Decree of the President of the Russian Federation, May 13, 2017, No. 208 "On the Strategy of Economic Security of the Russian Federation for the period up to 2030" (hereinafter -the Strategy), which comprehensively reveals the threats to Russia's economic security for near-term and the conditions for ensuring economic security. Conditions for ensuring economic security are the implementation of a set of political, organisational, socioeconomic, informational, legal and other measures aimed at countering and eliminating threats. As the main directions of ensuring economic security provided by the Strategy, there are two main ones: reducing the risks of doing business associated with the possibility of using formal reasons for stopping it or unjustified selective decisions regarding business entities; developing and taking measures for social protection of various segments of the population. The timely adoption of the document was confirmed by the measures taken by the Government since March 2020. This is confirmed by unprecedented measures to support small and medium-sized businesses, cityforming and system-forming companies, as well as social support for various segments of the population since the introduction of restrictive measures due to the spread of coronavirus infection in March 2020. Scientific studies conducted by NAFI experts show that almost half of the respondents are pessimistic about the prospects for normalisation of the situation in their industry, believing that the situation will normalise no earlier than in a year. About 31% believe that the situation can return to normal only in a few months. At the same time, entrepreneurs call first of all to postpone the payment of taxes or cancel them, to provide subsidies or financial support from the state, to introduce rental and credit holidays, issue interest-free loans to support business, as well as reduce or cancel mandatory contributions to the Social Insurance Fund. At the same time, in these areas of business, it was most necessary to restructure such business processes as personnel management; marketing; planning; online sales and several others. Support from the state was received by specific industries that are significantly dependent on consumer demand:  to support airlines affected by the cancellation of flights due to coronavirusmore than 23 billion rubles.;  for the development of domestic tourism -15 billion rubles;  subsidies to tour operators -3.5 billion rubles;  the program to stimulate demand in the automotive industry -25 billion rubles;  for the purchase of aircraft and helicopters for air ambulance -15 billion rubles;  for the program of subsidising loans to developers -12 billion rubles. Active measures to support citizens and businesses, primarily small and medium-sized businesses, provide not only confidence in the future, but also political stability. The receipt of significant support measures from the state in developed European countries at the initial stage of the crisis in march 2020 ensured the livelihoods of companies and the population. As the main directions of state support, the EU countries apply an increase in the amount of loans and deferred payment of interest, direct subsidies to small businesses, a ban on the termination of lease agreements. In Russia, business operations economic security and the reduction of threats to its liquidation, a significant reduction in jobs, in 2020 is ensured through the application of the following measures:  state aid;  concessional lending,  instalments and deferred tax payments;  instalment payment of debts when filing a claim until October 1, 2020;  prolongation of planned and audit audits;  automatic renewal of 15 types of licenses;  reduction of insurance premium rates;  extension of the terms of payment of administrative fines from 60 to 180 days for small and medium-sized businesses, except for fines for violation of traffic rules and trade in alcoholic beverages;  increase in advances on government contracts;  extension of the lease of state or municipal real estate. Pivotal measures to ensure the economic security of companys in 2020 are taken if business entities meet several conditions: Particular attention should be paid to the economic security of various categories of citizens in crises. Protection of the labour population in current difficult conditions is ensured by the preservation of jobs, the prevention of staff reduction by more than 10%, the provision of loans for up to 6 months at a 0% rate for the payment of the wages. Suppose you lose your job after March 1, 2020, as a result of the closure of companies due to coronavirus. In that case, you can remotely register as an unemployed person, pass free retraining programs, receive unemployment benefits from 4,500 rubles to a maximum of 12,130 rubles with an extension of payments for 3 months, payments of 3,000 rubles for each child. Measures are being implemented to support families with children through lump-sum payments, extending the terms of payment of benefits, changing the method of calculating family income when assigning social support, the possibility of disposing of maternity capital using electronic services, doubling the minimum child care allowance to 1.5 years. CONCLUSION To ensure the economic security of economic entity, a generic method to confront the threats do not exist and require the development of a comprehensive approach, including the definition of security, identifying security threats, developing threat protection, the formation of a security with a comprehensive approach to security management. In turn, an integrated approach to the system of ensuring the economic security of the company requires the formation of a multi-level structure with a comprehensive assessment of economic security at each level of company management. The average level allows you to assess the security of system objects that are previously grouped based on external economic threats. At the lower level, a set of measures should be formed to ensure the security of objects and structures of the company. The personal contribution of the authors lies in the synthesis of factors of external and internal threats of economic safety of an economic entity, the role definition information field and its organisations to collect relevant information to adopt measures for the sustainable financial state and future development and systematisation of measures of the state support of business in terms of 2020 corona-crisis. The study does not take into account all risk factors and only confirms one of the most critical trends in the system of economic securitythe need for a comprehensive assessment of the level of economic security and generalisation of support measures taking into account existing threats and risks.

Diet and Nutritional Status of Rural Preschool Children in the State of Orissa Abstract The data on diet and nutritional status of preschool children at State level is available, however the same is not available at district level for planning appropriate action programmes at that level. Hence in keeping view of this objective the present paper focused on to assess the diet and nutritional status of rural preschool children at district level in the State of Orissa. This study was conducted in rural areas of 30 districts in the State of Orissa. Dietary history was obtained from 1,605 preschool children and about 6,803 children (1-6 Yrs) were covered for anthropometry and clinical examination. The data revealed that the mean intakes of majority of foods except cereals and millets, roots and tubers and vegetables were below the recommended dietary intakes in both the age groups of preschool children. Similarly, in general, the intakes of majority of nutrients were below the RDA. The overall prevalence of Kwashiorkor and Marasmus was 1.1% each. The prevalence of Bitots spots was 2.7%. About 63% of preschool children were stunted and underweighed. Majority of preschool children was subsisting on inadequate diets and the diets were grossly deficient of micronutrients such as riboflavin, vitamin A and iron. The poor dietary intakes are reflected by higher prevalence of stunting and underweight.

This invention relates to a method for synchronizing the clock signal in a digital data receiver to decode pulse position encoded signals. In pulse position modulation (PPM) the time position of pulses carries the message to be communicated. The positioning of pulses is referenced to a clock signal which defines slot times in which signals are expected to appear. For example, at a clock frequency of 14.7 MHz, and using ten clock signals to define a slot, each slot time is 680 nanoseconds long. For a 32-slot PPM frame, a pulse appearing in the first slot would represent the number 0, a pulse appearing in the second slot would represent the number 1, and so on, counting upward to the last slot which would be the number 32, or in hexadecimal representation #1 F. Thus, we can see that five binary bits are encoded and transmitted in a 32-slot PPM frame. When receiving a PPM signal, it is necessary to decide in which slot an arriving pulse belongs, usually in "real time" as the pulses are received. If a pulse is incorrectly assigned to the wrong slot, due to variations in the time position of the pulse or the slot, the digital signal is incorrectly decoded. It is desirable to have a method for correctly assigning pulses to slots, and for correcting variations in the time positions of the slots by changing the phase or timing of the clock signal.

Development and validation of an automated solidphase extractionLCMS/MS method for the bioanalysis of fluoxetine in human plasma A widerange, specific, and precise liquid chromatography tandem mass spectrometric (LCMS/MS) technique for quantifying fluoxetine (FLX) in human plasma was developed using the RapidTrace ® automated solidphase extraction (SPE) method; the analyte and internal standard (IS) were extricated on Oasis MCX SPE cartridges. Acetonitrile and 5 mM ammonium formate buffer (90:10 v/v) were used as mobile phase to achieve chromatographic separation on the reverse phase (C 18 column). The analyte and IS were ionized using +ve electrospray ionization approach which was further traced by multiplereaction monitoring on a tandem mass spectrometer. To quantify the FLX and FLXd5, the parenttodaughter ion transition of m/z of 310.0/44.1 and 315.0/44.0 was used, respectively. The method demonstrated a linear active limit of 0.2030 ng/ml with recoveries ranging from 63.04% to 79.39% for quality control samples and 61.25% for IS samples. The concentrations over the calibration range demonstrated acceptable precision and accuracy. Due to the high inconsistency of the FLX concentration data, the minimum threshold of the assay was kept at 0.20 ng/ml. The flow rate was maintained at 500 L/min, and the time for sample analysis for each injection was 3.5 min. The method was found to be specific, sensitive, and faster with minimum utilization of organic solvents and was utilized further for metabolic and pharmacokinetic studies. analyzed by using the +ve ion electrospray ionization with the multiple-reaction monitoring (MRM) technique. The approach was validated following the USFDA Bioanalytical Method Validation Guideline. INTRODUCTION Fluoxetine (FLX) is mainly administered for the management of major depressive disorder (MDD). MDD is characterized by mood depression, losing interest in daily activities, altered cognitive functions, and deteriorating physical health, and these symptoms altogether result in a low quality of life. The pharmacotherapy of MDD utilizes FLX and aims to elevate the mood without causing significant side effects and to prevent the relapse of the diseases. Chemically, FLX "(R,S)-N-methyl-3-phenyl-3- propan-1-amine" is containing both R and S racemates in equimolar amount and categorized as an inhibitor of selective serotonin reuptake (SSRI). SSRIs are considered safer than other antidepressant drugs due Journal of Advanced Pharmaceutical Technology & Research | Volume 12 | Issue 3 | July-September 2021 to mild adverse effects, hence approved for pregnancy, teenagers, and children populations. FLX is metabolized by N-demethylation into active metabolite norfluoxetine in the liver. However, active metabolites' production takes a few days due to the longer half-life of the parent compound. Thus, it becomes challenging to measure the active metabolite in the plasma. It requires conducting studies with a longer duration, which adds to limitations as these studies involving human volunteers. As a result, most of the analytical studies rely on the measurement of FLX levels in biological fluids for pharmacokinetic and pharmacodynamic extrapolations. Several methods are available for the quantification of FLX, including the high-pressure liquid chromatographic (HPLC) methods with fluorescence detection, electrochemical detection, and gas chromatography, and HPLC along with MS detection. However, all the methods involve manual extraction of the compound of interest from the biological samples and subsequent chromatographic separation and quantification. The sample preparation process in the chromatographic analysis is considered a very crucial step. When it is manual, it makes the process very tiresome and time-consuming and leads to compromised final results with reduced precision and accuracy. In addition to compromised results, due to manual extraction, these methods have long run time, low sensitivity, narrow range, and require a large volume of biological samples. As per the available literature up to now, no methods are available for the quantification of FLX with this broad range and with automatic extraction process using the RapidTrace ® (Biotage) solid-phase extraction (SPE) unit. Here, we established a simple, fast, responsive, and robust tandem LC-MS/MS technique for the quantitative assessment of FLX in human K3EDTA plasma. An automated SPE system was used to increase the process accuracy and precision. Plasma concentrations of FLX were analyzed by using the +ve ion electrospray ionization with the multiple-reaction monitoring (MRM) technique. The approach was validated following the USFDA Bioanalytical Method Validation Guideline. Working standards and chemicals The reference working standards of FLX (98.71%) and FLX-d5 (99.9%) were procured from Clearsynth Ltd., India. Acetonitrile and methanol, liquid ammonia (25% NH 3 ), orthophosphoric acid, and ammonium formate were procured from Sigma-Aldrich, Germany. In-house preparation used Milli-Q water (Direct-Q WPS) in the study. Instrument and conditions Chromatographic/mass spectrometric condition Shimadzu HPLC was used to perform chromatographic separation on an Ascentis ® Express C18 reverse-phase column, which includes a pump (LC-10ADVR), column oven (CTO-10AVP), and an autosampler (SIL-HTC). The column oven was integrated with an autosampler; 10 L of extracted samples was directed to the column at Standard solution preparation The stock solutions of FLX and FLX-d5 were prepared in ACN and Milli-Q water (70:30 v/v) at 1 mg/ml. All the stock dilutions were prepared in diluent, water, and methanol (50:50 v/v) under the low-light condition and stored in refrigerator. The calibration curve and quality controls (QCs) were obtained by diluting a sufficient stock solution to obtain plasma concentration between 0.201 and 30.00 ng/ml and for IS as 50 ng/mL in the diluent. Sample preparation/extraction The automated SPE technique was opted to process the plasma samples using RapidTrace ® (Biotage) SPE system. The drug extraction from plasma samples was achieved through MCX cartridges (30 mg/cc). The extracted samples were evaporated to complete dryness at 45°C ± 2°C using a nitrogen evaporator (RapidVap Vertex evaporator). The dried extracts were reconstituted with mobile phase (500 L) and transfer into vials for further examination . Method validation USFDA guideline for analytical method validation was chosen to validate the method to test the selectiveness, linearity, precision, accuracy, matrix impact, and drug recovery. In addition to this, other validation parameters tested were analyte and IS stability, benchtop stability, benchtop stability during extraction, freeze-thaw stabilities, long-term stabilities, and standard stock solutions and reference solution stabilities. The reproducibility of the method was evaluated by determining reinjection reproducibility, dilution integrity, and ruggedness. RESULTS AND DISCUSSION For measuring the concentration of FLX in biological matrices, few HPLC, gas chromatography mass spectrometry (GCMS) and liquid chromatography mass spectrometry (LCMS) methods are available. These methods, however, have some drawbacks, therefore, LC-MS/MS approach for measuring FLX was considered by utilizing the RapidTrace ® automated SPE system, and detailed processes are given in Table 1. To create a reliable, consistent, and reproducible LC-MS/MS method chromatographic, separation conditions were optimized after MS/MS optimization to offer adequate separation and peak symmetry with an appropriate response. Chromatographic and mass spectrometric conditions are established in Table 2. The method was developed and validated keeping in mind a bioequivalence pharmacokinetic approach, in which healthy subjects were involved. The evaluation of parameters such as selectiveness, linearity, precisions and accuracies, drug recovery, stabilities (freeze-thaw, benchtop, long-term, and stock solution stabilities), and ruggedness was completed for the validation of the method. The selectivity was performed on different lots of hemolyzed and lipemic plasma samples. Insignificant interference was found in the various plasma sets tested at the RT of FLX and FLX-d5 . A graph of a representation of regression calibration curve is provided in Figure 2. The coefficient of variation (CV) of the experiment was calculated by using QCs as a % mark scale of sample concentration. Table 4 shows the precision and accuracy statistics for the interday and intraday periods. The chromatograms of FLX and FLX-d5 blank samples, blank + IS samples, limit of quantitation (LOQ) and upper limit of quantitation (ULOQ) are presented in Figures 3-6, respectively. The presence matrix effect will alter the ionization pattern of analytes present in the matrix. Thus, the technique was designed to evade any effect on the matrix. It was found that the absolute matrix effect was 98.4%, 98.5%, and 99.5% at lower, middle, and higher levels, respectively. This indicated that matrix effect in the presence of plasma was found to be negligible. The mean overall recovery of FLX was found to be 68.63%, with a precision of 13.58%. Whereas, the mean overall recovery of FLX-d5 was found to be 61.2%, with a precision of 4.7%. Table 5 shows the comparative recoveries and complete matrix effects with procedure proficiencies for FLX and FLX-d5. The stability was performed in both matrices, namely aqueous as well as plasma, and the results of the stability exercise are tabulated in Table 6. It is evident and apparent that analytes as well as IS stability samples were found to be stable under different storage circumstances. The QC samples were prepared to determine dilution accuracy. The nominal percentage of dilution integrity was found to be 94.9% and 94.2% after 2-4 times dilution, whereas the percentage of CV was stated to be 2.9 and 6.8%, respectively. The method showed ruggedness for the selected validation range . CONCLUSION In this method, a reverse-phase chromatographic method with tandem mass spectrometric detection using FLX-d5 as an IS was developed in the K3EDTA plasma. Chromatographic separation with a lower baseline was achieved using mobile phase with acetonitrile and ammonium formate buffer. The maximum response was observed using the reverse-phase Ascentis ® Express C18 column at an optimized flow rate (0.5 ml/min). This technique was established and found to be rapid, specific, precise, and linear and can facilitate the biostudies of FLX. Chromatographic and extraction parameters have been designed to achieve a short-run time and a fast, automated plasma sample processing procedure due to the chromatographic run duration of 3.5 min and retention time of 1.6 min. Thus, the method makes a high sample throughput. This approach can also be implemented in a clinical laboratory for therapeutic drug control, enabling individual dosage optimization, drug interaction identification, and patient compliance evaluation.

MUMBAI: The civic body’s internal report on the Himalaya foot over bridge crash is likely to corroborate the probable cause cited initially—welding failure. The final report, which was to be submitted to the municipal commissioner within a month of the crash on March 14, is still awaited. Civic sources said it may be submitted in a week. Seven people lost their lives in the tragedy. It may be recalled that recently the chief engineer of bridges had admitted at a civic meeting that the engineers did not “study” the structural audit report on the FOB adequately. The auditor had pointed to its deteriorating condition. The internal report will probe whether the audit report was studied and analysed properly by the municipal staff and action taken accordingly. In the immediate aftermath of the crash, a preliminary report identifying the municipal staff and structural auditor responsible for the incident was submitted to the civic chief. Further inquiry into the incident was ordered and a report sought in a month. Civic sources said there was a delay by the investigation team in procuring the documents on the FOB’s history, its maintenance and repairs. They said the debris of the bridge, carried to a site in Cuffe Parade, was tested to ascertain the cause of the sudden collapse. “As the FOB was constructed more than three decades ago, there might have been a welding failure which the civic staff responsible for the maintenance did not detect. The final report is likely to corroborate the fact,” said an official. Meanwhile, the government has appointed a committee to probe the incident and review all foot over bridges in the state. Manoj Saunik, principal secretary, PWD, confirmed it, saying, “We will suggest measures to be taken to maintain bridges.” It is expected to submit its report in three months.

Diagnostic decision. The measurement of anticonvulsant agent levels. Monitoring levels of antiepileptic drugs is useful in formulating a therapeutic plan but not in making a diagnosis. Differences in assay methods are not important. Differences in therapeutic ranges do not reflect procedural differences but merely the source that is quoted. Therapeutic ranges are not absolute criteria for therapy, but guides to a clinical balance between dose and side effects. Because therapeutic ranges reflect trough (early morning) serum levels, clinical determinations should be done at this time. Serum drug determinations should be used to establish the successful drug level for a patient; assess compliance; identify the relative contributions of each drug to efficacy and to side effects in complex programs; identify bioavailability problems; and characterize idiosyncrasies in the rate of drug metabolism.

Q: Basic Greek language resource for Philosophy purposes? Sometimes when I'm reading philosophy some greek terms become a problem (I'm looking at you, Heidegger), and I have to keep checking the Internet for translations and explanations. I've already searched for resources to learn Greek, but they are mostly focused on either learning the language as a whole, or learning traveling phrases. Also, they are usually about modern Greek, and I don't know how useful it would be regarding the (Ancient?) Greek used in Philosophy. I was trying to find something that, for example, focus on roots and how they combine so one can get a general understanding of terms that come up in philosophical texts. So, is there a good resource to learn basic Greek for Philosophy purposes? A: I use the Perseus Project for looking up Heidegger's greek quotations. It has greek and english word search, and paragraph-by-paragraph translations for many of the texts Heidegger quotes from. Very useful. http://www.perseus.tufts.edu/hopper/ Example screen-shot :-

This invention relates to an improved shift circuit which shifts data at a high speed. FIG. 1 is a schematic block circuit diagram of a central processing unit 2 (hereinafter called "CPU 2"). The CPU 2 is a single chip circuit comprising temporary registers (A, T, B, M) 4, an arithmetic logic circuit 8, a shifter 10, a microprogram ROM 12, a branch control unit 14, a status control unit 16, a common bus buffer/receiver 18, a timing generator & bus controller 22, and a special function unit 26. The temporary registers 4 store data supplied through an internal bus 6. The arithmetic logic unit 8 carries out arithmetic logic operations such as addition, subtraction, AND, OR with respect to data set from the temporary registers 4. The shifter 10 is designed to shift the data sent from the temporary registers 4. The microprogram ROM 12 stores information showing the sequence in which data are processed by the CPU 2. The branch control unit 14 controls the branching of a microprogram. The status control unit 16 includes flip-flop registers for storing the current status of the CPU 2 and a circuit for controlling the status of the flip-flop registers. The common bus buffer/receiver 18 receives and supplies data through an external bus. The timing generator & bus controller 22 controls transmission of data between the CPU 2 and a memory means or an input-output device and produces clock signals for defining the timing in which data are stored into the temporary registers 4 and general registers. The special function unit 26 is used to, for example, expand a bit arrangement. The shifter 10 is a conventional shift register. It shifts a 1-bit data at one cycle. Thus, to shift an m-bit data it would take m-cycle time. To shorten the time necessary for shifting data a new type of computer has been developed which has a shifter for shifting data at a high speed. Such a shifter is shown in FIG. 2. FIG. 2 is a block circuit diagram of a shifter consisting of a plurality of data selectors (DS) which are cascade-connected. The shifter receives an n-bit input data (n=8) and shifts the data by m-bit positions (m&lt;n) at one cycle. To the shifter an 8-bit data is supplied through input lines 28, a 3-bit data specifying an amount of shifting (i.e. m) is supplied through first control lines 30, and a 1-bit data specifying a "right" or "left" shift is supplied through a second control line 32. Both the 3-bit data and the 1-bit data are supplied to the data selectors DS. If the 1-bit data specifies "right" shift, the 8-bit data is shifted to the right by m bits and is obtained through output lines 34 at one cycle. If the 1-bit specifies "left" shift, the 8-bit data is shifted to the left by m bits and is obtained through output lines 36 at one cycle. With the shifter shown in FIG. 2, however, several steps must be carried out, as will be described below, in order to shift a multi-length data (2n bits, 3n bits, . . . ) by m bits either to the right or to the left.

The anti-arrhythmic eff ects of prednisone in patients with sarcoidosis Atrial fi brillation (AF) aff ects 2.3 million people in the United States and is currently the most common cardiac arrhythmia. Its overall prevalence is only increasing as the population ages. The classical risk factors for developing AF include hypertension, valvular disease, ischemic cardiomyopathy, and thyroid disease. In some patients with AF, an underlying cardiovascular pathology is not identifi ed and the etiology remains unknown. Treatment modalities for AF typically include rate control medications, antiarrhythmics and radio frequency ablation (RFA), each of which is accompanied by its own risk of complications. We report a case of symptomatic AF that was refractory to multiple antiarrhythmics and an RFA procedure which resolved with prednisone. In this case, AF was associated with cardiac sarcoidosis, a disorder that is thought to be due to granulomatous involvement of the myocardium and increased systemic infl ammation.

Extra-pulmonary tuberculosis: A retrospective study of patients in Accra, Ghana Background Information on extrapulmonary TB (EPTB) patients is limited in many African countries including Ghana. The study objective was to describe the epidemiology of EPTB patients diagnosed from different categories of health facilities in Accra, Ghana compared to pulmonary TB (PTB) patients and identify risk factors for mortality among EPTB patients. Method We conducted retrospective analyses of demographic and clinical data accessed from medical records of EPTB and PTB patients from different types of health facilities from June 2010 to December 2013. Factors at diagnosis associated with EPTB compared to pulmonary TB (PTB) and factors associated with treatment outcome death among EPTB patients were assessed using logistic regression. Results Out of 3,342 new TB patients ≥15 years diagnosed, 728 (21.8%) had EPTB with a male: female ratio of 1.17. The EPTB sites commonly affected were disseminated 32.8%, pleura 21%, spine 13%, and Central Nervous System (CNS) 11%. Treatment success rate for EPTB was 70.1% compared to 84.2% for PTB (p<0.001). In logistic regression, HIV positivity (adjusted Odds Ratio 3.19; 95% confidence interval 2.693.79) and female gender (aOR 1.59; 95% CI 1.351.88) were found to be significantly associated with EPTB compared with PTB. Older age, being HIV positive (aOR 3.15; 95% CI 1.208.25) and having CNS TB (aOR 3.88; 95% CI 1.1413.23) were associated with mortality among EPTB patients. While more EPTB patients were diagnosed in the tertiary hospital, health facility type was not associated with mortality. Conclusion EPTB patients in Accra have a worse treatment outcome compared to PTB patients with mortality of EPTB being associated with HIV, older age and CNS TB. Being HIV positive and female gender were found to be significantly associated with EPTB. Increased awareness of these factors may facilitate early case finding and better management outcomes for these patients. Introduction Despite major strides in prevention, diagnosis and treatment, tuberculosis continues to be a major leading cause of death globally. An estimated 1.67 million people died from TB in 2016. The causative organism Mycobacterium tuberculosis, which is predominantly airborne, affects the lung causing pulmonary TB. When TB is bacteriologically confirmed or clinically diagnosed in other parts of the body other than the lung such as the abdomen, meninges, genitourinary tract, joints, bones, lymph nodes and skin it is classified as extrapulmonary tuberculosis (EPTB). The prevalence of EPTB among new and relapse TB cases globally in 2016 was 15%. The lowest prevalence (8%) was recorded in the WHO Western Pacific Region while the highest (24%) was recorded in the Eastern Mediterranean. The figure for the African Region was 16%. The African Region, which has 13% of the world's population, accounted for 23% of the 918,011 EPTB cases reported globally in 2016. There is variation in the proportion of EPTB among TB cases in Africa with 5.2%, 31.7% and 41.3% reported for Nigeria and Ethiopia and Djibouti respectively. EPTB rates are even higher in the North African countries with Morocco and Algeria reporting 44.4% and 60% respectively. Ghana, a high HIV/TB burden country, with an estimated 28 million population reported 14,675 TB cases in 2016. The proportion of EPTB patients reported among new TB cases has been in the range of 8 to 10% over the period between 2006 and 2016. Even though data from the Ghana National TB Control Program (NTP) indicate that the prevalence of EPTB among TB patients has remained fairly stable there is concern about the mortality among EPTB patients which almost doubled from 7.8% in 2006 to 14% in 2012. The investigation of contributory factors to this mortality rate was highlighted as an area of interest for research in the NTP 2015-2020 Strategic Plan. Various risk factors reported to be associated with EPTB include immunosuppression, HIV infection, male gender and younger age. On the other hand, other studies have found females and increasing age to more associated with EPTB. The sites of EPTB vary by age group and gender across different populations studied, however lymph nodes and pleura invariably feature among the top reported sites from a myriad of studies. In view of the peculiarities of EPTB including the atypical nature of presentation, understanding the epidemiology of EPTB in diverse settings is of interest given the relevance of EPTB to TB control. Except for two teaching hospital setting studies on EPTB conducted in Ghana, studies on the EPTB in-country are limited. The current study therefore sought to highlight the epidemiology of EPTB patients diagnosed from different categories of health facilities in Accra compared to pulmonary TB (PTB) patients and identify risk factors for mortality among EPTB patients. initiative were described elsewhere. Accra, with a population of 1.7 million in the 2010 census, recorded HIV prevalence of 2.1% among antenatal clinic attendees over the period of 2010 to 2013. The facilities from which the study participants were derived included outpatient departments (OPD), HIV clinics and diabetes clinics in polyclinics, general hospitals, a regional hospital and a teaching hospital. The participants from the teaching hospital were from the HIV clinic only and did not include patients from the OPD or other clinics. These facilities accounted for 70% of TB cases in Accra at the time. From the TB case finding initiative database which consisted of 3,704 records, the participants for this study were selected using the following inclusion criteria: patients 15 years and older newly diagnosed with smear positive pulmonary TB, smear negative pulmonary TB or extra-pulmonary TB. The exclusion criteria were patients less than 15 years and those previously treated for TB. The classification of PTB and EPTB by the National Tuberculosis Control Program (NTP) in Ghana falls in line with WHO guidelines. With the exception of cerebro-spinal fluid (CSF) samples, which were usually quite small in volume, EPTB samples for microscopy and culture were taken through a decontamination process to get rid of other bacteria using a 4% sodium hydroxide (NaOh) and N-acetyl L-cysteine (NALC) preparation. An equal volume of NaOH-NALC solution was added to the sample for a quarter of an hour followed by neutralization with phosphate buffer solution pH 6.8. The preparation was then subjected to centrifugation for concentration of the specimen and to wash off the sodium hydroxide reagent. The decontaminated concentrated sample was then inoculated for culture and smear preparation. At the time of the case finding initiative, sputum smear samples for examination under light or light emitting diode (LED) microscopy were processed using Ziehl Nielsen staining method. Sputum smear positive PTB was defined as a patient with acid fast bacilli in at least one sample of sputum. A patient was considered to have sputum smear negative PTB if he or she had two sputum smears negative for mycobacteria on microscopy, but Chest X-ray showed evidence consistent with active tuberculosis. EPTB was classified as per organs or systems affected exclusive of the lungs, such as lymphatic comprising of TB in lymph nodes, pleura, spine, TB in bones and joints other than the spine, central nervous systems CNS (TB meningitis, brain), abdominal and other such as genito-urinary tract. EPTB diagnosis was based on having one culture-positive specimen using fine needle aspiration biopsy or organ fluid samples such as ascetic or pleural fluid depending on the suspected site involved, or histological evidence or strong clinical confirmation of active EPTB for which the clinician makes the decision to treat with a full course of TB drugs. Culture methods available included solid culture using Lowenstein-Jensen media and liquid culture by means of Bactec Mycobacteria Growth Indicator Tube (MGIT960, BD, Sparks, USA). These diagnostic methods were available at the teaching hospital laboratory which also performed culture on samples that were delivered from other lower level facilities. In the event that a patient has EPTB in several organs, the patient is classified according to the site that is most severely affected. In NTP registration, patients diagnosed with both PTB and EPTB were registered as pulmonary TB. It is therefore not possible to distinguish which patients had both types of TB. At the time of the case-finding initiative, the same standardized first line TB drugs were used to treat new cases of EPTB and PTB for the duration of 6 months. Classification of treatment outcomes were cure, treatment completed, default, died, transfer out and treatment failure as per WHO guidelines. The combination of those recorded as having been cured and completed treatment were designated as having a favorable treatment outcome. Data of study participants obtained from their medical records included age, gender, HIV status, type of TB, site of the EPTB, facility of diagnosis, year of diagnosis and treatment outcome. Data analysis was done by means of STATA version 12. The frequencies and percentages of the respective types of EPTB in totality and then stratified by gender and the age distribution were assessed. Multi-variate logistic regression analysis was conducted to identify factors associated with EPTB relative to PTB and risk factors associated with mortality among EPTB patients. Baseline characteristics including age group, gender, HIV status were included in the first model. In the second model those with who died during treatment were compared to those successfully finishing their treatment. Age group, gender, HIV status, affected site and facility of diagnosis were all included in the second model. Adjusted OR, 95% CI and p-values were calculated for each potential predictor variable with p-value of <0.05 set as the level of significance. Data collected for the analysis did not have any personal identifying information and was handled with strict confidentiality. Ethics approval for the study was obtained from the Ghana Health Service Ethical Review Committee. Results Out of 3,704 TB patients recorded in the TB case finding initiative database, 219 children less than 15 years and 143 patients who were not new TB cases were excluded from the analysis. The study participants consisted of 3,342 new TB patients who were aged 15 years and above. The overall male female ratio was 1.68. A total of 1,443 (42.9%) of these TB patients, were from the polyclinics; 775 (23.2%) were from the general hospital; 775 (23.2%) were from the HIV clinic of the teaching hospital and 359 patients (10.7%) were from the regional hospital. There were 728 patients (21.8%) who had extra-pulmonary TB while 2,614 patients had pulmonary TB. Out of the 728 EPTB patients, 400 (55%) were diagnosed from the HIV clinic of the teaching hospital. Almost half (48.5%) of the 2,614 pulmonary TB patients, were diagnosed in the general hospitals. For 646 (88.7%) of the EPTB patients, the site affected was recorded. Two hundred and twelve (32.8%) of the EPTB patients had the classification of disseminated TB, while for 134 patients (18.4%) the site affected was the pleura (Fig 1). Twenty patients (2.9% of EPTB cases) making up the category of "other" had various sites affected including pericardial, genitourinary, skin, and breast. Significantly more males 25.4% (87/343) reported pleural TB compared to females: 15.6% (47/302) p<0.01, while CNS TB was more common among females 15.2% (46/302) than men: 8.2% (28/343) p<0.01. For the other categories of EPTB, almost similar proportions of males and females were affected. Treatment outcome was documented for 665 (91.3%) EPTB patients. The overall treatment success rate was 70.1% for the EPTB patients (Table 1) with 6 patients reported to be cured. Treatment success ranged from 57.9% to 74.6% with 2012 being the year that treatment success rate was highest. The lowest treatment success rate was documented for 2013, possibly related to the low proportion of EPTB patients for which the treatment outcome was known (68%). Among EPTB patients, the overall mortality rate was 28.7%. The mortality rate was highest among those with CNS EPTB (52%) and disseminated EPTB (47%). Death rate among those with pericardial EPTB was also very high, but the number was very small (3/5 patients). Among PTB patients, the overall treatment success rate was 84.2% while the mortality rate was 12.6%. There was male predominance for both EPTB and PTB. For EPTB, the male: female ratio was 1.17:1 while for PTB, the male: female ratio was 1.87:1. The proportion of females with EPTB was significantly more than the proportion of females with PTB (p < 0.001) Table 2. The age distribution for both EPTB and PTB by gender is shown in Fig 2. Treatment success was significantly higher among PTB patients than EPTB patients. The HIV status was known for almost all the TB patients (98.8%). Among these patients for whom the HIV status was known, about two fifths (40.8%) were HIV positive. About 62% of EPTB patients were HIV positive compared to 32.4% of the PTB patients. The participants in the study from the teaching hospital came from the HIV clinic and as such all these TB patients were HIV positive. Among these HIV positive patients from the teaching hospital, 52% had EPTB. Among those who were HIV positive in the regional hospital, 20.5% had EPTB while the general hospitals and polyclinics had 8.3% and 6.3% respectively of the HIV positive patients having EPTB. The cure rate and treatment success rate for HIV positive EPTB was 1.3% and 51.8% respectively while the figures for HIV positive PTB patients were 26.8% and 66.7% respectively. The multivariate logistic regression analysis showed that EPTB patients were more often female and HIV positive compared to PTB patients ( Table 3). The likelihood of developing EPTB was more than 3 times among those with HIV. On the other hand, the age groups did not show any differential association with EPTB. Table 4 shows the results of a multivariate logistic regression with mortality as outcome compared to those treated successfully among EPTB patients. Risk factors for death included increasing age, HIV positivity and CNS TB. Going by the cut off for statistical significance the type of health facility attended by the EPTB patients did not appear to be associated with mortality, however mortality among EPTB patients attending the teaching hospital was worse than for the other facilities with the p value just falling shy of the level of significance at 0.077. Discussion This study elaborates on EPTB diagnosed among patients attending different types of facilities and HIV clinics in Accra over a period of three and a half years and compares demographic and clinical characteristics of these patients with those having pulmonary TB. One fifth of the newly diagnosed tuberculosis had EPTB. The most common form of EPTB was disseminated TB followed by pleural TB. A favorable treatment outcome was observed for seven out of ten EPTB patients with documented treatment outcome. Being HIV positive and female gender were found to be significantly associated with EPTB compared with PTB while older age, being HIV positive and having CNS TB was associated with mortality among EPTB patients. The reported proportion of EPTB among TB patients in our study (21.8%) fell within the range of what has been reported for other countries such as Swaziland (18.4%), Cameroon (19.4%) and Botswana (25%), which like Ghana are also classified as having a high HIV/TB burden. In this same category of HIV/TB burden countries, Uganda and Malawi have EPTB prevalence reported of 11% and 41%, respectively, highlighting the wide variation of the EPTB prevalence across countries and the uncertainty of reasons for this observation. Interestingly, the prevalence in our study was relatively higher than the countrywide data in the range of 8-10% reported for Ghana. It is worthy to note that our study population was derived from facilities in Accra and also consisted of patients from the country's premier teaching hospital HIV clinic, which has a large clientele. Usually such tertiary facilities have more sensitive diagnostic options for identifying EPTB. Secondly being a referral hospital, it receives a range of patients from other facilities including possibly HIV positive EPTB patients who may have been referred because of difficulties in making a diagnosis. This may explain the preponderance of the EPTB patients from the teaching hospital and the observed prevalence of EPTB. The HIV prevalence of 40% among our patients was also high compared to the 24% reported for Ghana because of the selection of the clinics for the case finding initiative which in the teaching hospital involved only the HIV clinic to the exclusion of the OPD and other clinics. Different studies report the pleura and the lymphatic system as the commonly affected extra pulmonary sites, while in our study disseminated TB topped the list followed by the pleura and other sites similar to what others have reported. The rate of disseminated TB that we found was similar to what was reported in a study of EPTB patients seen in a referral hospital in the capital of Cameroon. Disseminated TB is a severe form of EPTB usually associated with HIV infection with progression linked to the immuno- suppression and delayed treatment. Given the HIV coinfection in our study population and with disseminated TB presenting with systemic symptoms, it is possible that delayed diagnosis may have contributed to this magnitude of disseminated TB. In consonance with other studies, our study found an association between female gender and EPTB. While the reasons for this finding have not been clearly identified, it is suggested that genetic factors, gender differences in exposure to TB and the presence of other risk factors such as smoking could possibly be the linking factors. Further studies are needed to clearly delineate the linkages. The distribution of the site affected by EPTB across gender however does not appear to follow a clear pattern as various studies show similar or slightly different gender predominance in one site or the other. The predominance of males among those with pleural TB found in our study has also been reported by other researchers. Similar to what was observed in other studies, we found no association between EPTB and age. The treatment success rate for EPTB in our study was comparable to what Gomes et al. reports for EPTB patients in Brazil but lower than what was reported by studies conducted in Benin, Ethiopia and India. On the other hand, it is higher than what was reported by other researchers in Nigeria. Various systems of DOTS implementation may be contributory factors to the differences. The treatment success rate for Ghana has shown consistent improvement over the years with figures exceeding 85% over the course of the time during which data for our study was recorded. This achievement has been attributed to various factors including social and biomedical factors including the enablers package, community involvement in treatment and the used of fixed dose combinations. The EPTB patients in our study had a relatively lower treatment success rate than the national average for the country. Among those with poor outcomes, it was observed that death was responsible for an overwhelming majority. The rate of disseminated TB among our patients may be associated with the mortality observed given that in their study of mortality among Ghanaian TB patients, Nassikas et al. and Burton and colleagues showed disseminated TB as one of the highest risk factors for death. The HIV co-infection predominance among EPTB patients compared to PTB and the finding of HIV and CNS TB as risk factors for death in our study corroborates findings from different studies and highlights the importance of early initiation of anti-retroviral therapy for the survival of HIV-infected EPTB patients [7,10,29,. As pointed out by Ade et al. severe immune-suppression found in HIV patients with EPTB predisposes to these patients succumbing to opportunistic infection and ultimately death especially when anti-retroviral therapy is started late. This re-iterates the importance of vigilance to facilitate early diagnosis of EPTB to improve treatment outcomes and minimize the risk of progression to advanced forms and death. TB infection in people is mainly caused by Mycobacterium tuberculosis complex. The data on the causative pathogens for EPTB was however not available to this study. Unlike for pulmonary tuberculosis, very little has been studied on the Mycobacterium tuberculosis complex species responsible for EPTB in Ghana. One study conducted in Accra however found M. tuberculosis to be the predominant organism (eighty-eight percent of the samples) with M. africanum being the only other organism accounting for the remainder. In various studies on pulmonary TB in Ghana in which samples were analyzed, M. tuberculosis was also the major organism of M. tuberculosis complex isolated followed by M. africanum accounting for 73% to 97.6% and 2.4% to 23% of samples respectively. M. Bovis was however isolated in fewer studies and accounted for 0.4 to 3% of the samples. EPTB diagnosis in general poses a challenge given the problem of obtaining the appropriate specimen from suspected EPTB sites and the fewer Mycobacterium bacilli in EPTB samples limiting the identification of AFP using microscopy. At the time of the TB case finding initiative, microscopy which has low sensitivity but high specificity, was the main stay of TB diagnosis in these lower level facilities though further tests such as cultures on samples could be accessed at the teaching hospital laboratory. Fewer cases of EPTB were diagnosed in general hospitals and clinics in our study and may reflect a lack of appropriate well-defined diagnostic algorithms as available for PTB as well as inadequate facility resources and clinical expertise. Since generally more people access health care at lower level health facilities compared to tertiary hospitals, the cost benefit may be in favour of enabling these non-tertiary facilities using appropriate algorithms to freely access molecular tests and culture, which have higher sensitivity and specificity compared to microscopy. This may improve timely diagnosis of EPTB especially among those living with HIV to ultimately reduce morbidity and mortality. Our study shows that health facility type was not significantly associated with mortality. Further studies could explore possible factors at play including the association if any between the time to diagnosis and initiation of treatment at the respective facilities and treatment outcomes. Our study has some limitations. It involved secondary data analyses with the data source being the TB-case finding initiative database. Consequently there could be potential errors in the data since it was not possible to verify the diagnosis of EPTB or assess whether some of the study participants were misdiagnosed as EPTB. Similarly, the clinical improvement of patients who completed treatment could not be verified. Secondly, it was also not possible to distinguish patients who had concurrent EPTB and PTB among the study population as this information was not clearly indicated. This is against the background that in reporting as per Ghana NTP guideline, patients with both EPTB and PTB are recorded as PTB. There was also the challenge of incomplete documentation. The data from the teaching hospital consisted only of patients from the HIV clinic which may have introduced an element of bias in the results showing the association between EPTB and HIV. This result should therefore be interpreted against the light of this potential bias. Finally, with the study population being derived from Accra, the study findings may preclude generalization to the rest of the country. Despite these limitations, a major strength is that, to our knowledge, this is the first study that uses data from different facility types in Ghana to elaborate on EPTB in comparison to PTB, the treatment outcomes of EPTB patients and the risk factors for mortality. Conclusion This study in summary showed female gender and HIV co-infection as risk factors for EPTB, and HIV and CNS TB as risk factors for death among EPTB patients. Increased awareness of these factors, provision of and training in country-adapted diagnostic algorithms and making more sensitive diagnostic tools accessible may contribute to earlier case finding and diagnosis of EPTB patients especially at lower level health facilities for initiation of treatment and possibly better management outcomes.

Changes in peripheral immune cells after intraoperative radiation therapy in low-risk breast cancer Abstract A detailed understanding of the interactions and the best dose-fractionation scheme of radiation to maximize antitumor immunity have not been fully established. In this study, the effect on the host immune system of a single dose of 20 Gy through intraoperative radiation therapy (IORT) on the surgical bed in low-risk breast cancer patients undergoing conserving breast cancer has been assessed. Peripheral blood samples from 13 patients were collected preoperatively and at 48 h and 3 and 10 weeks after the administration of radiation. We performed a flow cytometry analysis for lymphocyte subpopulations, natural killer cells (NK), regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSCs). We observed that the subpopulation of NK CD56+high CD16+ increased significantly at 3 weeks after IORT (0.300.42%, P < 0.001), while no changes were found in immunosuppressive profile, CD4+CD25+Foxp3+Helios+ Treg cells, granulocytic MDSCs (G-MDSCs) and monocytic MDSCs (Mo-MDSCs). A single dose of IORT may be an effective approach to improve antitumor immunity based on the increase in NK cells and the non-stimulation of immunosuppressive cells involved in immune escape. These findings support future combinations of IORT with immunotherapy, if they are confirmed in a large cohort of breast cancer patients. INTRODUCTION Breast cancer is the most common malignancy and the second highest cause of death in women worldwide. The molecular mechanisms of breast cancer, particularly tumor cell immune evasion from the host immune response, have been the focus of intense research. The immune system appears to play an essential role in the initiation of breast cancer, progression and response to treatment, given that immune activation is associated with a higher pathological response. The current treatment for early-stage breast cancer is breast conservation surgery followed by radiation therapy. The tumor bed is a 110 preferential site of relapse. Increased local control is associated with improved overall survival, so events close to the tumor bed are crucial for the final prognosis. Several trials have evaluated various techniques of accelerated partial breast irradiation (APBI) of the lumpectomy bed vs conventional whole-breast irradiation in terms of efficacy and other outcomes. The results indicate that treatment with intraoperative radiation therapy (IORT) yields similar results in terms of survival and toxicity as whole-breast radiation therapy, with the benefit of reduced treatment time. In this context, IORT has been established as a good option for patients' treatment in early-stage breast cancer undergoing conserving surgery, or in combination with external beam radiation therapy (EBRT) in patients with risk factors. Research on the interactions between radiation therapy and the immune system has shown new mechanisms that can be exploited to improve the efficacy of radiotherapy. Radiobiology has focused on normo-fractionated schemes, but little is known about the biological effects of high doses per fraction. Radiotherapy can induce the release of antigens during the death of cancer cells that, in association with proinflammatory signals, activate the innate immune system by inducing the activation of specific T cells against the tumor. Radiotherapy also has effects on the tumor microenvironment, favoring the infiltration of activated T lymphocytes and overcoming some of the barriers of antitumor rejection. However, radiotherapy exerts immune inhibitory effects too. The limited availability of intratumoral dendritic cells to present radiation-released antigens and the increase in the irradiated tumor microenvironment of differents factors, including, e.g. transforming growth factor (TGF) and immnusopressive cells, such as myeloid-derived suppressor cells (MDSC) and regulatory T (Treg) cells, are part of theses immune inhibitory effects. Studies have shown that conventional fractional EBRT produces an increase in both the levels of MDSCs and Treg cells characterized by systematically expanding and promoting T cell dysfunction. However, it has been reported that the highest single doses applied in hypo-fractionated schemes are more advantageous for stimulating the immune system. Hypo-fractionated radiotherapy inhibits hypoxia and reduces the recruitment of immunosuppressive cells into primary tumors, generating a microenvironment with lower Programmed Death-Ligand (PD-L1), releasing the inhibition on cytotoxic CD8+ and boosting not only the local but also systemic anticancer immunity. This approach has been explored in radiotherapy techniques using high-dose fractionations such as stereotactic body radiotherapy (SBRT). IORT involves precise delivery of a large dose of ionizing radiation to the tumor bed, which is the area of higher risk of relapse. Although conclusive data on immune reactions and IORT are not available, it is likely that the single high doses administered during IORT cause immunogenic cell death of cancer cells remaining in situ after surgery by activation of the immune system. There are some published investigations on the effect of IORT on tumor cells as well as wound fluid. The results suggest that higher radiation doses result in increased tumor cell necrosis and antigen presentation, and recruitment of T-cells to irradiated and possibly distant unirradiated tumors, providing an additional reason to investigate IORT anew, perhaps in combination with immune checkpoint blockade treatments. Therefore, systemic functional changes in response to high doses of radiotherapy delivered in the tumor bed by IORT as a single treatment modality were investigated in this preliminary study. We assessed changes in peripheral blood immune cell composition after IORT on the surgical bed in patients with low-risk breast cancer. The information provided by this study could be useful to suggest new combinations and timings of IORT and immunotherapy in breast cancer patients. MATERIALS AND METHODS The pilot study protocol was approved by the Clinical Research Ethical Committee of the hospital (protocol code IORT-01, reference PR009/16), and written informed consent was obtained from all participants. From October 2016 to September 2018, a total of 13 patients diagnosed with low-risk breast cancer (T1N0M0 and T2N0M0) who were candidates for breast-conserving surgery and IORT were eligible to participate in the study. Inclusion criteria were as follows: age >50 years, histologically proven invasive ductal carcinoma <3.5 cm, low-risk luminal A subtype (estrogen-receptor and/or progesteronereceptor positive), HER2 negative, and protein Ki-67 <20%, without lymph node involvement (N0) adequate for breast-conserving surgery and IORT (lesion distance to skin or breast wall ≥0.7 cm), as well as capacity to understand the characteristics of the study and to give informed consent. Exclusion criteria were the presence of bilateral breast cancer at the time of diagnosis, a previous history of malignant disease except for non-melanoma skin cancer, need for treatment with chemotherapy, and state of immunosuppression, both current and developed during follow-up, caused by immunosuppressant medications or systemic disorders. For each patient, the postoperative pathological report was reviewed, and when high-risk factors were found, such as grade 3 tumor differentiation, lymphovascular invasion, triplenegative breast cancer subtype, positive nodes and affected margins or close margins <5 mm without re-excision, these cases were excluded from the analysis. Patients with these characteristics underwent EBRT with or without prior chemotherapy. Intraoperative radiotherapy was administered using the INTRA-BEAM system (Carl Zeiss Surgical, Oberkochen, Germany) emitting low-energy (50 kV) photons at a high dose-rate to the target tumor bed volume, with rapid dose fall-off and hence limited exposure to adjacent non-tumor tissues. Spherical applicators were used to deliver a uniform dose at the inner-surface of the breast lumpectomy cavity. The size of the applicator used in each case was based on the largest one that fits comfortably to the tumor bed so that the skin and subcutaneous tissue can be fixed with a suture on the bag over the sphere. The applicator was kept at least 0.7 cm from the skin. The dose rate varied according to the diameter of the applicator and the energy of the beam. A single dose of 20 Gy (in water) was prescribed and delivered in a period of 15-35 min, depending on the size of the applicator. IORT was administered immediately after removal of tumor volume when R0 resection was obtained. Flow cytometry Peripheral blood samples (9 mL) drawn in ethylenediaminetetraacetic acid (EDTA) were obtained preoperatively before surgery and IORT, and at 48 h and 3 and 10 weeks after IORT. Fresh blood was used, and the samples were processed within 24 h. Blood (50-100 L) of blood was added to the appropriate tubs, and cells were processed according to the manufacturer's instructions. The antibody panels used were the following: lymphocyte phenotyping panel ( All flow cytometry data were acquired on a 10-color/3-laser Gallios flow cytometer (Beckman Coulter). The instrument has not been altered. The stability of the flow cytometer was assured through a quality control procedure using Flow-Check Pro Fluorospheres (Beckman Coulter). A compensation matrix for each panel was created using the compensation tubes supplied with each panel, according to manufacturer's instructions. A minimum number of 100 000 leucocytes for lymphocyte phenotyping, 100 000 leucocytes for MDSC analyses and 40 000 leucocytes for regulatory T cell analysis was established. Data were manually analyzed using FlowJo software v. 10.5 (Tree Star Inc., Ashland, OR, USA). Statistical analysis Categorical data are expressed as frequencies and percentages, and continuous data as median and range. Friedman one-way Analysis of Variance (ANOVA) was used to compare changes of cell populations at different time periods and the Wilcoxon signed-rank test to assess differences in the cell populations at different time periods as compared to each baseline. Statistical significance has been confirmed by Conover post hoc test. Statistical significance was set at P < 0.05. Data were analyzed using the R statistical program (version 3.5.0). RESULTS A total of 13 women diagnosed with early-stage breast cancer were included in the study. The salient characteristics of these patients are shown in Table 1. The median age was 66 years (range 61-85 years). Most lesions were located in the upper outer quadrant (69.2%) of the left breast (84.6%). The most common form was the T1 stage (92.3%). In all patients, the histological diagnosis was infiltrating ductal carcinoma, luminal subtype. All patients received hormonal therapy. The most common size of the applicator was 3.5 cm (61.5% of the cases), with a mean treatment time of 19 min. No relapse was observed, and after a median follow-up time of 14 months, 100% of patients remained alive without disease. Immunophenotyping panel Of the 52 samples obtained at different time intervals in the course of IORT, 44 were analyzed. The flow cytometry gating strategy used for the analysis of all cell types is shown in Fig. 1. Lymphocytes were identified from CD45. T lymphocytes were distinguished from total lymphocytes using the characteristic CD3 marker. From these, CD4 + helper lymphocytes were separated from CD8 + cytotoxic. B lymphocytes were identified by characterization CD3 − CD19 +. To select the NK cells, a negative selection of the CD3 − CD19 − CD20 − markers was made; subsequently through the combination of CD56 and CD16, subpopulations of NK cells were obtained. Changes of total lymphocytes (CD45 + ) along the study period after the administration of IORT were not statistically significant (Friedman one-way ANOVA, P = 0.356). Also, differences in the paired comparisons (different time points vs baseline) were not statistically significant (Wilcoxon signed-rank test, P = 0.5, 0.95, 0.81 at 48 h, 3 and 10 weeks, respectively) ( Fig. 2A). When the different lymphocyte cell subpopulations were assessed, a progressive decrease in CD19 + B cells was observed from 48 h after IORT, reaching minimal values at 10 weeks (Friedman one-way ANOVA, P = 0.007). Although no statistically significant differences were found when the percentage of B cells in the baseline (7.38%) was compared with that found 10 weeks after IORT (5.91%), a trend towards significance was found (Wilcoxon signed-rank test, P = 0.062) (Fig. 2B). The subpopulation of T helper cells CD3 + CD4 + did not reveal any significant statistical differences among the different comparisons performed, although a slight decrease after IORT as compared to baseline (34.20%), reaching minimal values 10 weeks after irradiation (30.60%), was observed (Fig. 2C). By contrast, changes in CD3 + CD8 + cytotoxic T cells showed a progressive increase during the study Changes in peripheral immune cells after intraoperative radiation therapy in low-risk breast cancer 113 period, with the highest values at 10 weeks as compared with baseline (67.10 vs 60.20%), but these differences did not reach statistical significance either when the changes of cell populations at different time periods or when the different time periods were compared with the baseline (Fig. 2D). The CD4 + /CD8 + ratio showed an increase in cytotoxic lymphocytes from 0.56 to 0.45% at 10 weeks, but this change was not statistically significant (Wilcoxon signed-rank test, P = 0.58) (Fig. 2E). In the subpopulation of NK cells defined as CD56 dim CD16 +, there was a decrease from baseline to 48 h, but the difference was not significant. Differences between 3 and 10 weeks were not significant, although we observed a slight increase when compared with the baseline (Fig. 2F). In the population of NK cells with activated phenotype defined as CD56 +high CD16 +, differences along the study period after the administration of IORT were significant (Friedman one-way ANOVA, P = 0.007). When the different time periods were compared with the baseline there was a significant increase at 3 weeks compared with the baseline (0.30-0.42%, Wilcoxon signed-rank test, P = 0.001) (Fig. 2H). On the other hand, in the subpopulations of CD56 dim CD16 − and CD56 +high CD16 −, changes were not statistically significant in either of the comparisons by Friedman's test or the Wilcoxon's test for paired comparisons at different time points vs baseline ( Fig. 2G and I). In the subpopulation of CD56 − CD16 +, there was an increase from 13.20% at baseline to 15.80% at 10 weeks, but these differences were not statistically significant using Wilcoxon's test (P = 0.3125) nor for the comparisons at 48 h and 3 weeks. Also, differences were not significant when changes of cell populations at different time periods were compared (Friedman's test, P = 0.05) (Fig. 2J). Treg cells The strategy used for the analysis of different phenotypic and functional Treg subsets was screening lymphocytes according to side scattering and forward scattering characteristics, which were additionally blocked for CD4 + T cells. Then, CD4 + T cells were examined for CD4 + CD25 + Foxp3 + Helios + cell populations. Treg cells were also divided into functional subsets based on CD45RA and Foxp3 expression. The representative diagrams of flow cytometry for the analysis of all cell types are shown in Fig. 3. When changes in the cell populations at different timepoints were compared with the baselines, a slightly decreased CD3 + CD4 + CD25 + Foxp3 + Helios + cell population was observed at 48 h after the administration of IORT, with the lowest values at 3 weeks (27 vs 38.50% at baseline), although no significant differences were observed in any of the comparisons. Changes in the CD3 + CD4 + CD45RA − CD25 + Fig. 4. Median values of the CD4 + CD25 + Foxp3 + Helios + cell population (A) and CD4 + CD25 + Foxp3 + CD45RA − (B) during the study period at different time points. MDSCs Representative diagrams of flow cytometry for the analysis of all cell types are shown in Fig. 5. MDSCs were defined as CD45 + CD33 + CD -11b + cells. Then, MDSC cells were also divided into subsets based on CD14 and HLA-DR expression. Statistically significant changes in the overall MDSC population were not observed (Fig. 6A). For granulocytic MDSCs (G-MDSC) characterized as CD33 + CD11b + CD14 − and monocytic MDSCs (Mo-MDSC) characterized as CD33 + CD11b + CD14 + HLADR −/low, statistically significant differences were not found. Both in the G-MDSC and Mo-MDSC subsets, median percentages at different time points as compared with baseline were not statistically significant. Similar results were obtained when changes along the study period after the administration of IORT were assessed with Friedman's test (G-MDSC, P = 0.471; Mo-MDSC, P = 0.782) ( Fig. 6B and C). DISCUSSION Results presented in this study indicate that treatment with IORT in low-risk breast cancer patients undergoing conserving breast surgery can affect the balance of immune cells in the peripheral blood. During the study period, we noted that radiation therapy increased the presence of peripheral NK cells while no changes were detected in the immunosuppressive profile, characterized by Treg cells and MDSCs. Radiation therapy can cause apoptosis of T cells and lymphopenia. In the present study, statistically significant differences in the total number of peripheral blood lymphocytes were not found, but the population of B lymphocytes experienced a decrease after IORT, reaching its lowest values at 10 weeks. For the different subpopulations of T lymphocytes, a progressive increase in cytotoxic T lymphocytes throughout the study period was observed. The CD4 + /CD8 + ratio showed an increase of cytotoxic lymphocytes at 10 weeks as compared to baseline values. Differences, however, did not reach statistical significance probably due to the limited number of patients included in the study. Recent evidence suggests that T cells have an important function in regulating cancer growth. Several T cells play a critical role in the balance of anti-and pro-tumor immune responses within the tumor microenvironment as well as in the systemic circulation. In fact, cytotoxic T cells are the main final effectors of tumor cell death. However, Treg cells generally suppress or downregulate the induction and proliferation of cytotoxic cells. Therefore, the dominance of cytotoxic T lymphocytes is a favorable prognostic factor in cancer. We found significant changes in the subpopulation of NK cells in breast cancer patients treated with IORT. There is evidence that irradiation-induced tissue injury increases the expression of NK-activating ligands (e.g. NKG2D ligands) on malignant cells, thereby rendering tumors more susceptible to NK cell cytotoxic activity, modulating the immune response by increasing antigenicity and upregulating the expression of proinflammatory cytokines and chemokines. CD56 dim NK cells have a high cytotoxic activity and express high levels of perforin; in addition, the high expression CD16 makes them efficient mediators of antibody-dependent cellular cytotoxicity. CD56 + CD16 − NK cells have a lower cytotoxic capacity but are the most efficient cytokine producers, including interferon- (IFN- ), tumor necrosis factor- (TNF-), Interleukin-12 (IL-12), IL-15 and granulocyte and monocyte colony-stimulating factor. In the present study, the subset CD56 dim CD16 + NK cells showed an increase after 3 weeks of IORT and continued to increase until the end of the follow-up period (46.20-55.80%). The CD56 − subset expressing CD16 at high density, but showing functional alterations due to its low cytotoxic activity and cytokine production, presented variations during the study period. Although there was a decrease at 48 h, the percentage of cells increased at 3 and 10 weeks. We observed that the only significant increase was in the subset CD56 high CD16 + NK cells, especially at 3 weeks (0.30-0.42%). CD56 high CD16 + NK cells are characterized by NKG2A and low levels of perforin, and are primarily specialized for cytokine production. In peripheral blood these NK cells express CD62L, CCR7, CXCR4 and CXCR3 that allows their preferential recruitment to secondary lymphoid organs, tumors and inflamed tissues. One of the factors responsible for the variations in the percentage of these cells during our study period could be the presence of cytokine release, especially the overexpressed TGF- molecule after irradiation. In vivo studies in breast cancer patients have shown a positive correlation of TGF-1 with NKG2A receptor expression, thus suggesting that this molecule plays a role in these regulatory mechanisms. These findings indicate that a single radiation dose might increase the number of these cells that have antitumor capacity. The identification of NK cell subsets endowed with particular functional capabilities might help monitor residual antitumor NK cell-mediated responses in breast cancer patients. Treg cells can promote tumor progression by inhibiting effective antitumor immunity. These cells are more resistant to ionizing radiation than conventional CD4 + T cells and show a relative increase after radiotherapy. In our study, the percentage of peripheral blood CD4 + CD25 + Foxp3 + Helios + Treg cells did not experience significant change during the study period. This relevant finding indicates that the use of this single high-dose approach may be essential to neutralize these immunosuppressive cells, which are one of the main components of immune escape. In recent years, there has been great interest in promoting the production of more efficient Treg cells enhancing antitumor activity. In this respect, assessment of Treg cell levels and related cytokines in the peripheral blood may be important to guide individualized treatments in breast cancer. It has been shown that the prevalence of MDSCs with immunosuppressive activities is an important mechanism of tumor escape. In mouse models, accumulated evidence has demonstrated the importance of MDSCs in the development and progression of breast cancer. Gonda et al. examined MDSC levels in 155 patients with breast cancer. MDSCs in circulating peripheral blood increased in patients with breast cancer compared to healthy volunteers. Also, the levels of MDSCs in preoperative patients and in patients with recurrent breast cancer were significantly higher compared with postoperative patients, patients with recurrent breast cancer who received chemotherapy and healthy volunteers. Studies have shown that RT, especially conventional fractional RT, produces an increase in the levels of MDSCs characterized by being more resistant to radiation. However, in our study, changes in the overall MDSCs population as well as in G-MDSC and Mo-MDSC subpopulations were not observed. This finding is in line with the results previously published by our group in patients with lung cancer. We also observed that the use of high doses in hypofractionated schemes, in this case stereotactic body radiotherapy (SBRT), did not produce an increase in the percentage of MDSCs when analyzed in peripheral blood. Indeed a significant decrease in the percentage of granulocytes was observed 6 months after the administration of radiation. These results may show the possible benefit of the application of this type of scheme, which seems to be more effective in controlling cells related to immunosuppression. In this respect, studies have been developed to test if hypofractionated dose schedules may be more effective than conventional fractionated radiotherapy. In an experimental study of mice bearing murine melanoma treated with up to 15 Gy radiation given in various-size fractions, Schaue et al. found that fractionated treatment with medium-size radiation doses of 7.5 Gy/fraction gave the best tumor control and tumor immunity while maintaining low Treg cells number. In a preclinical study, Lee et al. compared a single 20 Gy dose with 5Gy x 4 over 2 weeks. Ablative 20 Gy radiation dramatically increased T-cell priming in draining lymphoid tissues, leading to reduction/eradication of the primary tumor or distant metastasis in a CD8 + T cell-dependent fashion, whereas fractionated radiation therapy showed a lower inhibition of tumor growth. However, the best dose-fractionation regimen to maximize antitumor immunity remains to be established. In this preliminary study, we show that the administration of a single dose of IORT altered the balance of peripheral immune cells by increasing NK cells but did not produce changes in Treg cells or MDSCs. No previous studies have evaluated the effect of IORT on peripheral immune cells in breast cancer patients. We therefore consider that these results help to clarify the interactions of IORT with the host immune system and add evidence to suggest IORT utilization as an attractive alternative to whole-breast irradiation in selected patients with early-stage breast cancer. However, given the exploratory nature of our study, more studies are needed, with a greater number of patients and a control group of patients receiving hypofractionated wholebreast irradiation, to support the use of the combination of IORT with immunotherapy as a new therapeutic option in breast cancer patients. SUPPLEMENTARY DATA Supplementary data is available at RADRES Journal online.

The Nonaligned and the United Nations. Edited by M. S. Rajan, V. S. Mani and C. S. R. Murthy. [Oceana keeping, disarmament, law or economic developmentare bland in their account of the UN, and optimistic about its potential. There is no serious analysis of why the problem of disarmament has proved so intractable, not just between the superpowers, but also in the post-colonial world. Nor is there deep discussion of whether decolonisation has itself led to new tensions and to refugee flows. The authors show little awareness of some of the defects of the UN system, or of the critique made in the United States by the Heritage Foundation. Where they do succeed is in giving a flavour of how, in Third World States, the UN is valued because it helped to create, and now symbolises, an international order free from the scourge of colonialism, and offering the hope of further progress in the direction of peace and equality. The Dutch volume, Peace and Security: Justice and Development, is the record of a congress held in Nijmegen. As a conference, it had merit: some excellent points were made. However, as a book it does not succeed: the texts of very general opening remarks, and edited summaries of debates, are not a substitute for thorough and reflective full-length analyses.

GRAND JUNCTION, Colo. (AP) — A U.S. judge says a Mexican national suspected of a 1989 slaying in Colorado can be put on trial in the United States, even though he was acquitted of the killing in Mexico. The Grand Junction Daily Sentinel newspaper reported Sunday (http://bit.ly/2qNNsOx ) that the judge ruled 67-year-old Rafael “Shorty” Aguilar Garcia can be tried in Colorado without violating the U.S. Constitution’s prohibition of double jeopardy. Garcia was suspected of killing Charles Porter but returned to Mexico before he could be arrested. The Mexican government is reluctant to extradite citizens charged with serious crimes in the U.S., so it put Garcia on trial. He was acquitted in 2012. Colorado authorities learned last year that Garcia had returned to the state and arrested him. A trial is scheduled this year.

The Value of Copper Sulphate in Umbilical Granuloma a Prospective Study Original Research Article Background: Umbilical granuloma is one of the commonest presentations in the pediatric surgery outdoor. Persistent drainage or moisture involving the umbilicus is chief complaint. If umbilical granuloma remains untreated, it could ooze and present with persisting irritation for several months. The main concern has been over avoidance of surgery in infants with umbilical granuloma. Different protocols have been proposed from time to time. Objective: To evaluate the therapeutic effect of copper sulphate on umbilical granuloma in infants. Materials and Methods: This prospective study was conducted on 25 infants with umbilical granuloma. In all patients Copper sulphate applied once over the Umbilical granuloma for 5 minutes. A sterile gauze and adhesive bandage were applied after cleaning of copper sulphate application. Treatments were administered in outpatient settings, and the patients were followed up at 1 week, and 2 weeks interval to record the outcome. Result: All 25 cases responded well to this approach with complete resolution of lesions. No major complication or recurrence was noted in follow-up. Conclusion: The use of copper sulphate in treating umbilical granuloma is simple, cost-effective, curative, and safe. INTRODUCTION Umbilical granuloma has always been a matter of parenteral concern and constitutes a common reason to attend the pediatric surgery outdoor. Umbilical granuloma is a benign abnormality of the neonates occur as a result of too much granulation tissue after separation of cord from the umbilicus. It also occurs as a reaction to a mild infection. Following the infection or cord separation, a tiny portion of a slightly wet, bright red, flesh remains in the umbilical cord. Failure of epithelialization of this left-over granulation tissue leads to the formation of umbilical granuloma. If not treated on time it may lead to infection, oozing, discharge, and irritation. A number of different modalities available for the treatment of umbilical granuloma such as cauterization with silver nitrate (most commonly used), common salt, electrocauterization, cryotherapy, alcohol, and ligation of the granuloma. Each of these methods has its merits and demerits, and some may have significant side effects, such as reports of skin burns following the use of silver nitrate. MATERIALS AND METHODS This prospective study was conducted on 25 patients attending the pediatric surgery outdoor of SMS medical college Jaipur, Rajasthan, India from September 2017 to April 2018 with a diagnosis of umbilical granuloma. In all patients Copper sulphate applied once over the Umbilical granuloma for 5-10 minutes. A sterile gauze and adhesive bandage were applied after cleaning of copper sulphate application. Treatments were administered in outpatient settings, and the patients were followed up at 1 week, 2 weeks, and 1-month interval to record the outcome. Complete regression, complete epithelialization, and no discharge interpreted excellent response while partial regression/epithelialization and no regression represented no response. RESULT All the patients were similar in terms of socioeconomic and ethnic profile. A total of 25 infants of aged 2-16 weak were included in the study . Fourteen infants were boys and eleven were girls . The effect of copper sulphate was evaluated 1 week, 2 weeks and 1 month following the last application. In 21 (84%) patients complete epithelialization occur in 5-7 days and 3 (12%) patients responded in 8-10 days. In one patient need second visit for copper sulphate application. All 25 (100%) infants demonstrated excellent results . DISCUSSION Umbilical granuloma is relatively very common surgical entity in infants at our center. Umbilical Granuloma develops due to failed epithelialization and increase growth of granulation tissue at the umbilicus after separation of umbilical cord. It may become complicated and led to the formation of pus for several months if left untreated, or not treated properly. In the literature, natural regression of the untreated umbilical granuloma has been not documented, so treatment is necessary. Different approaches have been suggested in literature such as chemical cauterization, electrical cauterization, cryo-cauterization, and treatment with topical copper sulfate, and common salt. Small granulomas can be treated with silver nitrate remedy while larger ones may require surgical resection. Chemical cauterization, with 75% silver nitrate was the most commonly used treatment. Use of silver nitrate in umbilical granuloma has been drastically cut down due to its propensity to cause burns if not well administered. Currently, electric cauterization or surgery is an option in failed noninvasive treatments. These treatments are surgical removal of the granuloma under general anesthesia. A disadvantage is that it costs more than noninvasive treatments. Copper sulphate is a suitable agent for the treatment of umbilical granuloma in this situation. It is potent and cost-effective, shows no adverse effects, and easily available. In our study, copper sulphate had a high response rate (100%) without any recurrence. In our study, the majority of the patients with umbilical granuloma were under the age of 8 weeks. This is consistent with the study of Anapurna et al. they reported the prevalent age group is less than two months in 82.14%. A study by Saleh A et al. also reported that the prevalent age group is less than two months is 72.0%. Present study has attempted to simplify the approach towards umbilical granuloma. It is to be emphasized that no surgery should be advised in these young patients. Proper counseling of the parenteral anxiety is important. In our study, copper sulphate, had a high response rate (100%) without recurrence. This finding is consistent with the observations of Annapurna et al. who found an excellent response with copper sulfate group (95.5%) and slightly better response to the common salt group (55%). they suggest that copper sulfate may be a more efficacious form of therapy than the common salt. CONCLUSION Our results showed that Copper sulphate is a reasonable method for Umbilical granuloma, resulting in good cosmetic appearance and acceptable complication rates. However, further randomized controlled study on large number of patients with longer follow up period is recommended to prove the therapeutic effect of copper sulphate in management of umbilical granuloma and in minimizing the need for surgery and recurrence.

Efficient and Fine-Grained VMM-Level Packet Filtering for Self-Protection In Infrastructure-as-a-Service (IaaS) clouds, stepping-stone attacks via hosted virtual machines (VMs) are critical for the credibility. This type of attack uses compromised VMs as stepping stones for attacking the outside hosts. For self-protection, IaaS clouds should perform active responses against stepping-stone attacks. However, it is difficult to stop only outgoing attacks at edge firewalls, which can only use packet headers. In this paper, we propose a new self-protection mechanism against stepping-stone attacks, which is called xFilter. xFilter is a packet filter running in the virtual machine monitor (VMM) underlying VMs and achieves pinpoint active responses by using VM introspection. VM introspection enables xFilter to directly obtain information on packet senders inside VMs. On attack detection, xFilter automatically generates filtering rules based on packet senders. To make packet filtering with VM introspection efficient, we introduced several optimization techniques. Our experiments showed that the performance degradation due to xFilter was usually less than 16%.

Improved survival after intraoperative cardiac arrest in noncardiac surgical patients. The article "Improved Survival After Intraoperative Cardiac Arrest in Noncardiac Surgical Patients" by Girardi and Barie 1 in the January issue of theArchivesis a worthwhile addition to the few articles on the subject that address the issue via a survey of large numbers of cases from a single institution over a relatively long time span. 2 The Invited Commentary by Flint 3 overlooks the fact that, after correction for type of operation, the ratio of cardiac arrest to number of operations has not changed appreciably over the last 35 or 40 years. The central problem is that while anesthetic agents and adjuvant drugs have changed, one invariant has remained, the so-called anesthesia record. Although it is true that during anesthesia one observes the patient, important decisions are not made on the basis of any one instantaneous observation but rather on the basis of the information accrued from the

This invention relates generally to the field of merchandising containers suitable for packaging small articles, clothing in folded condition, and similar merchandise, and more particularly to an improved form thereof in which provision has been incorporated to provide for the sealing of the carton to prevent undetected pilfering during such periods when the carton is in the custody of persons other than the purchaser after packing of a purchase has been completed. While the incorporation of pilfer-proof seals in large containers containing commercial shipments is well-known in the art, such provision normally involves the employment of additional structure incorporated into the main body and lid of such containers, the additional cost of which is amply justified by the value of the shipment, and the fact that the carton is, in many instances, reuseable. However, in recent years, the problem of pilferage occurring where a shopper checks articles he has already purchased, to be retrieved before completing a shopping expedition has been increasing. On the increase is pilfering occurring in restaurants, or checking facilities in a store and the like. Part of the problem exists in the fact that normally the design of a package or carton used for merchandising is almost entirely based around parameters of low cost and easy assembly from a flattened condition.

Commentary: listening can be exhausting--fatigue in children and adults with hearing loss. Anecdotal reports of fatigue after sustained speech-processing demands are common among adults with hearing loss; however, systematic research examining hearing loss-related fatigue is limited, particularly with regard to fatigue among children with hearing loss (CHL). Many audiologists, educators, and parents have long suspected that CHL experience stress and fatigue as a result of the difficult listening demands they encounter throughout the day at school. Recent research in this area provides support for these intuitive suggestions. In this article, the authors provide a framework for understanding the construct of fatigue and its relation to hearing loss, particularly in children. Although empirical evidence is limited, preliminary data from recent studies suggest that some CHL experience significant fatigue-and such fatigue has the potential to compromise a child's performance in the classroom. In this commentary, the authors discuss several aspects of fatigue including its importance, definitions, prevalence, consequences, and potential linkage to increased listening effort in persons with hearing loss. The authors also provide a brief synopsis of subjective and objective methods to quantify listening effort and fatigue. Finally, the authors suggest a common-sense approach for identification of fatigue in CHL; and, the authors briefly comment on the use of amplification as a management strategy for reducing hearing-related fatigue.

Single subcutaneous administration of RGDK-lipopeptide:rhPDGF-B gene complex heals wounds in streptozotocin-induced diabetic rats. Development of effective therapeutics for chronic wounds remains a formidable clinical challenge. Deficiency of growth factors is of paramount importance among the multitude of factors contributing to the pathogenesis of diabetic wounds. Clinical interest has been witnessed in the past for exogenous applications of platelet derived growth factor B (PDGF-B) in chronic nonhealing wounds. However, accomplishing even modest favorable clinical effects in such topical applications requires large and repeated doses of PDGF-B proteins. Chronic wounds are being increasingly circumvented by gene therapy approach and to this end, cationic liposomes are emerging as promising nonviral carriers for delivering various growth factors encoding therapeutic genes to wound beds. However, as in case of topical application of growth factors, all the prior studies on the use of cationic liposomes in nonviral gene therapy of wounds involved repeated injections of cationic liposome:cDNA complexes over several weeks for ensuring complete wound healing. Herein, we show that a single subcutaneous administration of an electrostatic complex of rhPDGF-B plasmid, integrin receptor selective RGDK-lipopeptide 1 and cholesterol (as auxiliary lipid) is capable of healing wounds in streptozotocin-induced diabetic Sprague-Dawley rats (as model of chronic wounds). Western blot analysis revealed significant expression of rhPDGF-B in mouse fibroblast cells transfected with RGDK-lipopeptide 1:rhPDGF-B lipoplex. The transfection efficiencies of the RGDK-lipopeptide 1 in mouse and human fibroblast cells preincubated with various monoclonal anti-integrin receptor antibodies support the notion that the cellular uptake of the RGDK-lipopeptide 1:DNA complexes in fibroblast cells is likely to be selectively mediated by alpha5beta1 integrin receptors. Findings in the histopathological stainings using both hematoxylin and eosin (H & E) as well as Masson's Trichrome staining revealed a significantly higher degree of epithelization, keratization, fibrocollagenation and blood vessel formation in rats treated with RGDK-lipopeptide 1:rhPDGF compared to those in rats treated with vehicle alone.

Tuberculous Addisons disease with increased hydrocortisone requirements due to administration of rifampicin A 58-year-old man was referred to our hospital for darkened skin, general fatigue and weight loss. His adrenocorticotropic hormone and cortisol levels indicated a primary adrenal insufficiency. 18Fluorodeoxyglucose positron emission tomography/CT showed bilateral enlargement of the adrenal glands, with 18fluorodeoxyglucose accumulation. Loop-mediated isothermal amplification assays of bronchoalveolar lavage fluid were positive for Mycobacterium tuberculosis. The patient was diagnosed with tuberculous Addisons disease and treated with antituberculosis agents, including rifampicin. The patients fatigue worsened gradually after initiation of rifampicin, and the dosage of hydrocortisone was increased. Serum cortisol level monitoring at 2hours after administration of hydrocortisone was shown to be clinically useful for determining the optimal dose, especially with concurrent use of rifampicin.

Holly Willoughby is fast cementing her status as a TV style icon. The This Morning presenter shared a photo to Instagram on Monday of her latest outfit, consisting of a cornflower blue cashmere jumper from Massimo Dutti, a knee-length leather skirt from Jigsaw, which highlighted her slim waist, and black LK Bennett court shoes. Holly (36) has avoided sharing the secret behind her weight loss since debuting a slimmer figure at the Glamour Women of the Year Awards earlier this year. Speaking to ITV's Lorraine Kelly, Holly simply said that having more time to herself allowed her to lose weight. "My children are a bit older, you get to that point where you have a bit more time to yourself," she said, not revealing how she lost the weight. "But I think as long as you're happy and healthy, that's most important." When asked about losing her "Willough-boobies," as dubbed by Keith Lemon, Holly (36) said: "I think they're still there, Lorraine." The mother-of-three also shared that having more children may be on the cards in the future. "I love being pregnant, I love giving birth, I love all that, it's like wearing a wedding dress, I don't want that all to be over. Maybe one day [we'll have more]? We're very happy as we are, it feels like we are complete." Holly recently celebrated her 10th wedding anniversary with Dan Baldwin, with whom she shares Harry (eight), Belle (six) and Chester (two).

Computed tomography of the normal soft tissues of the wrist. Computed tomography and sectional radiography and photography were performed on eight cadaveric wrist specimens in order to define the soft tissue structures of the normal wrist. Three specific levels were analyzed in detail: the levels of the distal radioulnar joint, of the pisotriquetral joint, and of the hook of the hamate. All soft tissues, with the exception of arteries and veins, were well demonstrated with computed tomography. In particular, the contents of both Guyon's canal and the carpal tunnel could be delineated. The position of the extensor carpi ulnaris tendon and sheath could also be evaluated. Contrast opacification of the flexor tendon sheaths and palmar synovial sacs did not contribute to the analysis. Computed tomography appears to be an important imaging modality in the evaluation of significant clinical problems, including the carpal tunnel and ulnar tunnel syndromes and subluxation of the extensor carpi ulnaris tendon.

A $3.5 million international bribery scheme involving oil pipelines in Siberia has landed a Pennsylvania-based businessman in federal prison. And a panel of the U.S. Court of Appeals for the Third Circuit has just ruled that Dimitrij Harder should stay there. His 5-year prison sentence is justified, despite Harder’s claim that his graft had an economically beneficial result for residents of that part of the former Soviet Union, Judge Eugene E. Siler concluded in the circuit court’s opinion. For starters, Harder, 46, is a German citizen, a Russian émigré and a permanent U.S. resident. Investigators said he paid the bribes to an official of the multinational European Bank for Reconstruction and Development in 2008 and 2009. The bribes were to speed approval of financing for clients of Harder’s who wanted a piece of the pipeline projects, officials said. At the time, Harder was an international financial consultant and president of the Chestnut Consulting Group in Southampton, Bucks County. Harder pleaded guilty in 2016 to violating the federal Foreign Corrupt Practices Act, which forbids U.S. businesses from using payola to obtain projects overseas. U.S. Eastern District Judge Paul S. Diamond slapped Harder with the prison term, plus a $2 million forfeiture order, after Harder argued he should receive leniency because “there was no intended victim and no actual victim,” Siler noted. On appeal, Harder claimed Diamond didn’t give enough credence to that contention. Siler found that Diamond did consider Harder’s argument and rightly rejected it. Diamond also noted at Harder’s sentencing hearing that one of Harder’s clients who benefited from the bribery was prosecuted in England and sentenced to 6 years in prison.

HIF-2 Suppresses p53 to Enhance the Stemness and Regenerative Potential of Human Embryonic Stem Cells Human embryonic stem cells (hESCs) have been reported to exert cytoprotective activity in the area of tissue injury. However, hypoxia/oxidative stress prevailing in the area of injury could activate p53, leading to death and differentiation of hESCs. Here we report that when exposed to hypoxia/oxidative stress, a small fraction of hESCs, namely the SSEA3+/ABCG2+ fraction undergoes a transient state of reprogramming to a low p53 and high hypoxia inducible factor (HIF)-2 state of transcriptional activity. This state can be sustained for a period of 2 weeks and is associated with enhanced transcriptional activity of Oct-4 and Nanog, concomitant with high teratomagenic potential. Conditioned medium obtained from the post-hypoxia SSEA3+/ABCG2+ hESCs showed cytoprotection both in vitro and in vivo. We termed this phenotype as the enhanced stemness state. We then demonstrated that the underlying molecular mechanism of this transient phenotype of enhanced stemness involved high Bcl-2, fibroblast growth factor (FGF)-2, and MDM2 expression and an altered state of the p53/MDM2 oscillation system. Specific silencing of HIF-2 and p53 resisted the reprogramming of SSEA3+/ABCG2+ to the enhanced stemness phenotype. Thus, our studies have uncovered a unique transient reprogramming activity in hESCs, the enhanced stemness reprogramming where a highly cytoprotective and undifferentiated state is achieved by transiently suppressing p53 activity. We suggest that this transient reprogramming is a form of stem cell altruism that benefits the surrounding tissues during the process of tissue regeneration. INTRODUCTION Numerous studies have revealed that although the majority of transplanted stem cells die, and do not engraft, stem cells secrete growth factors and antioxidants that likely contribute to the repair and regeneration of the inflamed and damaged tissues. This view is consistent with the findings that undifferentiated stem cells are capable of secreting high levels of antioxidants and growth factors. However, the site of injury/inflammation is particularly characterized by hypoxia and production of reactive oxygen species (ROS). The ROS could induce DNA damage, and subsequently trigger p53 activation in stem cells, leading to differentiation and apoptosis. Hence, it is suggestive that stem cells may have evolved specific mechanisms to maintain an undifferentiated state in the area of injury to serve their cytoprotective activity. Therefore, we posited that stem cells might have acquired molecu-lar mechanisms to suppress p53 and remain functionally undifferentiated during hypoxia and oxidative stress in order to be able to conduct their cytoprotective activity. One of the potential ways that stem cells could suppress p53 is by activating HIF-2a. Hypoxia upregulates the key transcription factors HIF-1a and HIF-2a with numerous downstream targets that play critical roles in survival, angiogenesis, metabolism, and other important functions. HIF-1a stabilizes p53, and therefore enhances p53 activation, whereas HIF-2a is shown to suppress p53 activity by reducing ROS generation. HIF-2a also upregulates several genes involved in antioxidant mechanisms including SOD, catalase, GSPx, and ABCG2. ABCG2 is an efflux pump, which is found to secrete glutathione (GSH), a primary antioxidant and component of the GSH/glutathione-s-transferase (GST) detoxification system. HIF-2a also enhances the secretion of growth factors having cytoprotective and regenerative abilities, for example, vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF)-2. Thus, one potential mechanism of p53 suppression would be to enhance HIF-2a expression, which could not only contribute to protect the stem cells from p53-induced differentiation or death but also enhance their cytoprotective activity. Therefore, we sought evidence that stem cells may suppress p53 activity during hypoxia and oxidative stress by upregulating HIF-2a in order to exert their cytoprotective and regenerative activities. We also wanted to determine whether the same mechanism might also be involved in the potential transformation of stem cells to cancer stem cells as has been recently documented To investigate the above possibilities, we used human embryonic stem cells (hESCs) as a model cell type. There are several advantages in using ESCs to test our hypothesis. First, p53 is highly active in hESCs and second, it directly inhibits Nanog, one of the core regulators involved in maintaining the stemness state. Third, HIF-2a is active in ESCs. We exposed hESCs to an extreme environment of hypoxia and oxidative stress and applied a novel approach to isolate a surviving, undifferentiated hESC fraction having low p53 and high HIF-2a activity. We were successful in isolating a small fraction of hESCs, the SSEA3/ABCG2 fraction, which exhibited high HIF-2a and low p53 activity as well as enhanced cytoprotective activity. While investigating the undifferentiated state, we serendipitously discovered that the SSEA3/ABCG2 cells undergo a transient state of reprogramming to an even higher state of stemness, here termed the ''enhanced stemness'' state, having very high Nanog expression, high antioxidant secretion, and high cytoprotective activity following exposure to hypoxia and oxidative stress. hESC Culture and Induction of Hypoxia Due permissions were obtained to culture hESCs both at Department of Medicine, Stanford University (Stem Cell Research Oversight protocol #285, director: BD) and at Department of Laboratory Medicine and Pathobiology, Hospital for Sick Children. BGO1 and H9 hESCs (WiCell Inc., Madison, WI, www.wi cell.org) were expanded on Mitomycin-C (10 lg/ml; Sigma)-inactivated mouse embryonic fibroblast (MEF) feeders, cultured in knockout Dulbecco's modified Eagle's medium/F12 with 20% knockout serum replacement (KOSR, #12660012, and #10828-028; Invitrogen Life Technologies, Grand Island, NY), and 4 ng/ml FGF-2 (#PHG0024, Gibco, Life Technologies, Grand Island, NY, www.invitrogen.com) as described elsewhere. The expanded hESCs were then grown on feeder-free Matrigel (##354230, BD Biosciences, San Jose, CA, www.bdbio sciences.com)-coated dishes with MEF-derived conditioned media (CM), supplemented with 8 ng/ml FGF-2 (complete hESC media), and under standard conditions (37 C and 5% CO 2 and saturated humidity) as previously described and used to perform various experiments described in the text. To make singlecell suspension for flow cytometry and to perform plating efficiency experiments, the hESCs were dispersed with TrypLe select (#12563-011, Invitrogen) for 3 minutes and then washed twice with the complete hESC media supplemented with 10 lM Rhoassociated protein kinase inhibitor Y-27632 (Calbiochem, Merck, Rockland, MA, www.emd.us). The hESCs were then suspended in the complete media plus Rho-associated protein kinase (ROCK) inhibitor to maintain viability during flow cytometry sorting. For the hypoxia experiments, day-2 hESCs culture having 30%-40% confluence was used and hypoxia was achieved as described previously. Briefly, hypoxic condition (<0.1% O 2 ) was established in a sealed chamber using the BBL GasPak plus anaerobic system envelopes with a palladium catalyst (Becton Dickinson, Cockeysville, MD, www.bd.com/ds). Immediately after hypoxia, the cells were incubated in tissue culture incubator at 37 C and 5% CO 2. Medium was changed only 24 hours after hypoxia so that the cultured cells were exposed to ROS generated immediately during the reoxygenation process. Flow Cytometry Flow cytometry was performed using FACScan and FACSAria II (BD Bioscience) as described. Surface staining and flow cytometry sorting was done as described elsewhere. Briefly, the single-cell suspensions of hESCs were incubated with primary antibodies (Supporting Information Table 1) in staining media (phosphate buffered saline (PBS) with 2% KOSR, Invitrogen), on ice, for 15 minutes, washed thrice in staining medium, and then incubated with Alexafluor secondary antibody for 30 minutes. After washing, the stained cells were suspended in complete culture media and kept at 4 C. Immunostaining Cells were fixed in 4% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA, www.emsdiasum.com) for 10 minutes on ice, permeabilized, washed thrice in PBS, and incubated overnight in staining buffer (4% bovine serum albumin (BSA) 0.1% saponin) with primary antibodies against HIF-2a and p53 (Supporting Information Table 1). They were then washed and incubated with secondary Alexa 488 or 594 antibodies for 1 hour at RT. For immunostaining of teratoma tissues, frozen sections were fixed with 4% paraformaldehyde, permeabilized with 0.1% saponin, and subjected to immunostaining as described elsewhere. Cell Viability and Apoptosis Measurement The hESCs were grown onto Matrigel-coated dishes in complete hESCs media. At day 2 after plating, cells were exposed to hypoxia for 24 hours and then reoxygenation for next 4 days. On day-4 post-hypoxia, the viability was determined by Trypan blue assay. A portion of the cells was subjected to flow cytometry sorting, and then the sorted population was subjected to caspase-3 activity assay as previously described. ELISA Assay The cell lysates were subjected to ELISA assay as previously described. Information about various ELISA kits used in the study is given in Supporting Information Table 1. InCell Western Assay The assay was performed as per manufacturer instructions (Licor Bioscience, NE, Lincoln, NE, www.licor.com). The hESCs (5 10 3 per well) were plated in sterile 96-well plates (BG Falcon, #353075) and kept at RT for 30 minutes before transfer to the incubator to reduce excessive settling at the well edges. The plates were incubated overnight at 37 C and then fixed with 3.7% formaldehyde at RT for 10 minutes. After fixation, the plates were washed with PBS, then primary antibodies were added at 1:300 concentration in 2% BSA 0.1% saponin, and incubated overnight at 4 C. After primary antibody incubations, plates were washed thrice (5 minutes each) with 100 ll per well PBS-T (PBS with Tween 20) at RT. Then, secondary antibodies conjugated to IRDye 800CW (goat-anti-rabbit-IgG (Licor Biosciences, NE) or goat-anti-mouse-IgG (Licor Biosciences, NE) were used at 1:1,000 dilution. The nuclear staining dye DRAQ5 (Biostatus Ltd., Leicestershire, U.K., www.biostatus.com) combined with Sapphire700 (Li-COR) at 1:10,000 and 1:1000, respectively, were also added. Plates were then incubated with 50 ll per well secondary antibody solutions for 90 minutes at R/T in the dark. Background control wells were prepared by omitting primary antibodies (i.e., secondary only). After secondary antibody incubations, plates were washed thrice times (10 minutes each) with PBS-T at R/T in the dark, and then plates were dried in air before scanning. Plates were scanned and analyzed using an Odyssey IR scanner using Odyssey imaging software 3.0. Scan settings used were medium or high image quality, 169 lm resolution, intensity 5.0 for the both the 700-channel and 800-channel with an offset of 4.0 mm. For signal quantification, antibody signals were analyzed as the average 800-channel integrated intensities from triplicate wells normalized to the 700-channel signal integrated intensity to correct for well-to-well variations in cell number. Results are expressed as fold change (means 6 standard errors of the mean) compared to noninfected controls. Measurement of Transcription Factor Activity The transcriptional activity of HIFs and p53 were measured by a transcription factor (TF) filter play assay system developed by Signosis, Sunnyvale, CA, www.signosisinc.com (Supporting Information Table 1). The assay was performed as per manufacturer instructions. Briefly, the nuclear extracts of hESCs were mixed with a specific biotin-labeled TF DNA binding sequence to obtain TF-DNA complexes. Then, the TF-DNA complexes were retained in a filter plate and hybridization was achieved. The captured DNA probe was quantified by streptavidin-horseradish peroxidase (HRP) method on a microplate luminometer. The Oct-4 and Nanog transcriptional activities were measured by Cignal Lenti Nanog reporter (Luciferase) kit (SABioscience, Valencia, CA, www.sabio.sciences.com) as per manufacturer instructions. Measurement of Oxidative Stress The level of ROS was measured by DCFH-DA assay as described. The oxidative DNA damage was measured by a 8-hydroxy-2-deoxy Guanosine (8-OH-dG) Enzyme Immuno Assay kit (#589320; Cayman Chemical, MI, Ann Arbor, MI, www.cayman chem.com) as per manufacturer's instructions. siRNA Gene Silencing We performed small interfering RNA (siRNA) gene silencing by Accell siRNA (Thermoscientific Dharmacon, Lafayette, CO, USA, www.dharmacon.com). The following Accell siRNAs were used: the human HIF-2a (A-004814-17-0005), HIF-1a (A-004018-24-0005), and p53 (A-003329-22-0005). The hESCs (5 10 3 per well in 96-well plate) were cultured in the completed hESC media, and 1 lM Accell siRNA was delivered to each well as per manufacturer instructions. After incubation for 72 hours at 37 C, the media were replaced with fresh media, and the cells were exposed to hypoxia. The gene knockdown was confirmed by the real-time quantitative reverse transcription-polymerase chain reaction (QPCR) and ELISA. Cytoprotective Assays For the in vitro cytoprotective assay, primary rat cardiomycotes were obtained from Cell Applications Inc. (San Diego, CA, www.cellapplications.com) and grown in the rat cardiomyocyte growth medium as per manufacturer's instructions. SH-SY5Y neuroblastoma cells were obtained from ATCC. The cells were exposed to 24 hours of hypoxia followed by reoxygenation for next 24 hours, and the viability was measured by Trypan blue assay. The cell culture medium was replaced with hESCs CM 2 hours before exposing to hypoxia. The in vivo bone marrow (BM) stem cell cytoprotection experiments were carried out in BALB/c mice as described previously. Briefly, 10-12-weekold BALB/c mice (Charles River Laboratories International, MA) were injected with hESCs CM i.p. 3 hours before the i.p. injection of high-dose 120 mg/kg carboplatin. On the day 5 of injection, mice were sacrificed, BMs were collected, and both the hematopoietic stem cell (HSC) and mesenchymal stem cell (MSC) fractions were isolated by using the easy-sep magnetic sorting kits (#19756 for HSC and #19771 for MSC; Stem Cell Technologies, BC, www.stemcell.com). The isolated cells were then subjected to colony forming unit (CFU) assay for HSCs and MSCs as described elsewhere. Limiting Dilution Assay for Teratoma Formation The in vivo teratoma study was performed by injecting hESCs mixed with 1:2 Matrigel to non-obese diabetic severe combined immunodeficient mice subcutaneously as described. The number of hESCs injected s.c. to mice were as follows: 2 10 6, 1 10 5, 1 10 4, 1 10 3, 1 10 1, and the number within parentheses representing number of animal used for the experiments. Mice were observed for 1-6 months for the teratoma formation. To confirm teratoma formation, the xenografts were fixed in formalin and subjected to H&E study. In a separate experiment, a portion of Matrigel plugs were removed 3 weeks after implantation to mice as described and frozen fixed to perform immunostaining for Nanog expressing cells by using a anti-Nanog antibody (Supporting Information Table 1). Statistical Analysis The statistical analysis was performed using GraphPad Prism 4.0 (GraphPad Software Inc., CA, www.graphpad.com) using either Student t test or one-way analysis of variance with Newman-Keul post hoc test. The flow cytometry data were analyzed by flowjo (Tree Star, Inc., OR). The analysis of teratomagenic efficiency was done by extreme limiting dilution software (http://bioinf.wehi.edu.au/software/elda) P53 and HIF-2a Are Upregulated in hESCs Following Hypoxia/Oxidative Stress To investigate whether exposure to hypoxia followed by reoxygenation (i.e., oxidative stress) activates p53 as well as HIF-2a in ESCs, BGO1, human ESCs were exposed to extreme hypoxia (pO 2 $ 0.1% for 24 hours) followed by reoxygenation (Supporting Information Fig. 1a), and changes in ROS, p53, and HIFs were measured. We found that BGO1 cells underwent marked morphological changes of differentiation and cell death (Fig. 1A) with an accompanying fourfold increase in ROS level (Fig. 1B). While the p53 protein level was increased fourfold by day 4 (Fig. 1B), HIF-2a protein was already rapidly elevated threefold after 1 day post-hypoxia (not shown) and remained sustained during the day-4 reoxygenation period (Fig. 1B). In contrast, HIF-1a protein expression increased immediately during hypoxia (day 0) and then showed a rapid reduction in expression by day-4 post-hypoxia (Fig. 1B). These results suggest that activation of both p53 and HIF-2a is sustained on day 4 during the reoxygenation phase following hypoxia. In contrast, HIF-1a decreased during the reoxygenation phase, which is consistent with the previous findings that HIF-1a mRNA is unstable whereas HIF-2a mRNA could be sustained for a prolonged period after hypoxia. The High HIF-2a and Low p53 Cells Are Enriched in the SSEA31/ABCG21 Fraction We next considered that the high HIF-2a cell fraction might be enriched within the undifferentiated ESC fraction and would express ABCG2, since this is one of the target genes of HIF-2a. To investigate this possibility, we first evaluated whether the high HIF-2a expressing hESCs on day-0 and day-4 post-hypoxia treatment overlapped with ABCG2 expression. We found that most of the HIF-2a expressing cells were positive for ABCG2 (Supporting Information Fig. 1b). Second, we sorted by flow cytometry a surviving (propidium iodide negative) undifferentiated SSEA3 fraction expressing ABCG2. The flow sorting showed that only 12% 6 3% (p <.05; n 4) SSEA3 cells survived by day-4 post-hypoxia/ reoxygenation (Fig. 1C), which was a eightfold reduction in the SSEA3 fraction (p <.05; n 4). We then found that the ABCG2/SSEA3 fraction (Fig. 1C) increased by fourfold ( Fig. 1C; 4.2 6 1.1, p <.05; n 4) by day-4 post-hypoxia/ reoxygenation. Third, we compared the HIF-2a protein levels as well as transcriptional activities between the SSEA3/ABCG2 and SSEA3/ABCG2 fractions using several assays. Immunofluorescence labeling revealed a strong expression of HIF-2a in the ABCG2 fraction versus the ABCG2 fraction (Fig. 1D), which was confirmed at the protein level as measured by ELISA (Fig. 1E), and at gene expression and the transcriptional activity level (Supporting Information Fig. 2a, 2b). The HIF-1a expression was not sustained in the day-4 post-hypoxia/reoxygenation ABCG2 fraction ( Fig. 1E; Supporting Information Fig. 2a). Taken together, our results suggest that HIF-2a expression was enriched in the SSEA3/ABCG2 fraction as compared to the SSEA3/ABCG2 fraction. Next we compared the protein level and transcriptional activity of p53 in SSEA3/ABCG2 versus SSEA3/ ABCG2. Immunofluorescence labeling indicated a low expression of p53 in the ABCG2 fraction versus the ABCG2 fraction (Fig. 1D), which was also confirmed quantitatively by ELISA (Fig. 1E), and by transcriptional activity as measured with the transcriptional filter assay (Supporting Information Fig. 2b). These results suggest that the SSEA3/ ABCG2 fraction expresses a low level of p53 as compared to the SSEA3/ABCG2 fraction. We reasoned that a high HIF-2a and low p53 activity could be associated with higher survival and less DNA damage in the SSEA3/ABCG2 fraction versus the SSEA3/ABCG2 fraction. Hence we measured levels of ROS, 8-OHdG, and caspase-3 and found these to be four-to fivefold higher in the SSEA3/ABCG2 versus the SSEA3/ABCG2 fraction (Supporting Information Fig. 2c). The trypan blue viability assay showed a 3.5-fold higher survival of SSEA3/ABCG2 versus SSEA3/ABCG2 (p <.05; n 4; data not shown). Thus, we conclude that the SSEA3/ABCG2 fraction was characterized by higher survival and less DNA damage as compared to the SSEA3/ABCG2 fraction. We then investigated whether the high HIF-2a/low p53/ ABCG2 phenotype generated through hypoxia/reoxygenation hereon designated ABCG2hox could persist in vitro for a prolonged period of time. We found that this cell fraction actively proliferated in the ESC medium for several weeks (three passages; total of 25 days). During this period, p53 and HIF-2a levels were measured by ELISA. The protein levels were compared with day-0 post-hypoxia/reoxygenation ABCG2 cell fraction. We found that the post-hypoxia/reoxygenation ABCG2hox fraction showed a sustained phenotype of low p53 and high HIF-2a expressions that lasted for 20 days (Fig. 1F). Taken together, we conclude that hypoxia/reoxygenation endows a subfraction of BGO1 ESCs with enhanced survival and is associated with high HIF-2a and low p53 activities. This high HIF-2a and low p53 phenotype could persist in vitro for more than 2 weeks and was mainly confined to the ABCG2 fraction. ABCG21Hox Cells Exhibit a Highly Undifferentiated and Cytoprotective State To investigate whether the ABCG2hox fraction maintained an undifferentiated stemness state throughout the 3 week culture period of low p53/high HIF-2a expression, we measured the flow cytometry expression and transcriptional activity of Oct-4 and Nanog in the ABCG2hox fraction and compared it with ABCG2nox (designating the SSEA3/ABCG2 fraction maintained under normoxia and not exposed to hypoxia/reoxygenation). First, we found that the levels of Oct-4 and Nanog protein expression measured by flow cytometry were sustained in ABCG2hox over the 2 week period (Fig. 2). Second, the transcriptional activity of Oct-4 and Nanog were sustained three-to fourfold higher in ABCG2hox versus ABCG2nox over the 2 week period (Fig. 3A). Thereafter the transcriptional activity level returned to a baseline level corresponding to the transcriptional activities of Oct-4 and Nanog in the ABCG2nox fraction by day-20 of reoxygenation (Fig. 3A), and similar to the period of high HIF-2a/ lowp53 state of ABCG2hox as previously described in Figure 1F. This result suggested that the ABCG2hox fraction, having a sustained high HIF-2a activity, also sustained a higher Nanog and Oct-4 transcriptional activity over the 2 week culture period. Consequently, we speculated that the ABCG2 protein level might also be sustained at a higher level in the ABCG2hox fraction for a correspondingly similar period. Previous studies have shown that the ABCG2 efflux pump confers a cytoprotective effect on ESCs by reducing intracellular ROS. A recent study reported that ABCG2 also secretes GSH, a potent cellular antioxidant, into the extracellular space. We, and others, reported that GSH exerts a cytoprotective activity on mammalian cells, including BM stem cells, against hypoxia/ reperfusion as well as cancer chemotherapy-induced toxicities. Hence, a corresponding increase in GSH secretion by the ABCG2hox fraction might confer cytoprotection on nearby cells exposed to ROS-induced toxicity. We investigated these possibilities as follows. First, we measured the ABCG2 protein level in the ABCG2hox versus ABCG2nox fractions by ELISA assay. We found a sustained threefold increase of ABCG2 expression in the ABCG2hox versus the ABCG2nox fraction (Fig. 3B). Second, we measured GSH secretion in the CM obtained from the ABCG2hox and ABCG2nox fractions with a GSH quantitative assay that is very sensitive for measurement of GSH in stem cells. We found a fivefold increase in the GSH level in the CM obtained from the ABCG2hox fraction (Fig. 3B). We also noted that the level of ABCG2 and GSH returned to the baseline level on day 20 corresponding to the return of the high-HIF-2a/low p53 state to its baseline state as in the ABCG2nox phenotype (Fig. 3B). Thus, it appears that the ABCG2hox phenotype is associated with high GSH secretion into the CM. Third, we compared the in vitro cytoprotection potential of the ABCG2hox-versus ABCG2nox-derived CM. CM was collected from 1 10 6 ABCG2hox cells in 4 ml, on day-8 post-hypoxia, and used to treat cardiomyocytes and neuronal cells (SH-SY5Y cell line ) exposed to hypoxia/ reperfusion, and was compared to nonexposed medium as a control. We then assessed the effect on survival and found a 10-12-fold increase in the survival of the cardiomyocytes and neuronal cells when treated with the CM of ABCG2hox versus CM from the ABCG2nox fraction (Fig. 3C). Thus we demonstrated that the ABCG2hox fraction CM exerted a significant cytoprotective activity on differentiated cardiomyocytes and neuronal cells, suggesting a mechanism whereby the ABCG2hox fraction is cytoprotective. To obtain in vivo evidence of the cytoprotective activity of the ABCG2hox-derived CM, we used a murine model of carboplatin-induced BM stem cell toxicity assay developed in our laboratory. In this assay, antioxidants were shown to protect BM HSCs and MSCs from high-dose carboplatininduced toxicity. We pretreated carboplatin-exposed mice with CM derived from ABCG2hox compared to CM from ABCG2nox. The CM were collected from 1 10 6 ABCG2hox and ABCG2nox cells in 2 ml, on day-8 posthypoxia, and 1 ml was injected i.p. to mice. We then measured both HSC and MSC survival using established assays. The CM was compared to N-acetyl-cysteine (NAC) a potent antioxidant, the latter having been shown to cytoprotect stem cells. We found that the CM derived from the ABCG2hox cells showed a 17-fold cytoprotection of HSC and 10-fold protection of MSCs (Fig. 3D). Interestingly, this cytoprotective activity in CM was roughly equivalent to NAC. Thus, we conclude that the ABCG2hox CM showed potent cytoprotective activity in vivo against BM toxicity induced by carboplatin, a commonly used anticancer agent in clinics. Taken together, our observations above suggest that the ABCG2hox fraction attains a highly undifferentiated state (henceforth known as enhanced stemness state) and exhibits potent cytoprotective activity. Considering the present interest in the potential use of hESCs for tissue regeneration, the results obtained from the BGO1 CM are provocative. To evaluate whether CM derived from the ABCG2hox fraction of other hESC lines could also exhibit similar regenerative potential, we repeated the above experiments with the H9 hESC line and obtained similar results. These results are described in Supporting Information Figure 3 and suggest that the H9 ABCG2 hox cells Figure 1D and maintained in the human embryonic stem cells (hESCs) culture media for next 3 weeks. In specific interval of time, the ESCs were fixed and subjected to flow cytometry expression of Nanog and Oct-4 as described. The inset panel at the top of the prehypoxia panel indicates the isotype control. The BGO1 hESCs were fixed and permeabilized before subjecting to incubation with primary and secondary (Alexa 488 for Oct-4 and phycoerythrin for Nanog) antibodies. Results shown are representative of a typical experiment (n 4; data from 5,000 single-cell events). exhibited a high HIF-2a, Oct-4, Nanog and low p53 phenotype, and exhibited in vivo cytoprotective activity comparable to that of NAC (Supporting Information Fig. 3). Thus, we conclude that the ABCG2 hox fraction in both the BGO1 and H9 hESCs exhibited cytoprotective activities. We then decided to evaluate the characteristic of the ABCG2hox phenotype using the BGO1-derived ABCG2hox cells. The Enhanced Stemness State Portends an Extreme Degree of Self-Sufficiency We noted that the ABCG2hox fraction showed a higher level of Nanog activity (Figs. 2, 3A). Since very high Nanog expression leads to an embryonal carcinomatous (EC) like state of extreme self-sufficiency, we investigated whether the ABCG2hox fraction attained extreme selfsufficiency. First, we assayed for the expression of FGF-2 and Bcl-2, involved in growth and regulation of apoptosis and associated with extreme self-sufficiency of ESCs and EC cells. We found that FGF-2 and Bcl-2 expressions were upregulated two-to threefold in the ABCG2hox versus ABCG2nox fractions (Fig. 4A). Second, we performed a standard clonogenic assay, minus supplementation with a ROCK inhibitor, on flow-sorted ABCG2hox and ABCG2nox cells grown on Matrigel as single cells, and the ESC colony formation was measured after 4 days. We found that the ABCG2hox cells showed a 10-fold higher colony forming capacity than the ABCG2nox cells (Fig. 4B). The colony forming efficiency was similar to that of Tera-2 cells (Fig. 4B). Our results are consistent with previous reports that high Nanog and Bcl-2 levels enhance the ability of flow-sorted single hESC to survive without the addition of the ROCK inhibitor. Thus, we suggest that the ABCG2hox cells demonstrated a significantly greater degree of self-sufficiency than the ABCG2nox fraction in the in vitro clonogenic assay. Next, we performed the in vivo teratoma assay to further characterize the enhanced state of self-sufficiency. The ABCG2hox and ABCG2nox were injected subcutaneously into immunocompromised mice to obtain teratomas (Supporting Information Fig. 4), and a serial dilution assay was performed to compare the teratomagenic frequency between the ABCG2hox and ABCG2nox. Tera-2 was used as a control. We found a 15-fold increase in teratomagenic efficiency of ABCG2hox versus ABCG2nox (Fig. 4C) and comparable to the Tera-2 cells (Fig. 4C). We noted that the ABCG2hox fraction contained a eightfold higher level of Nanog expression (Fig. 4D, and detail quantification given in the Supporting Information Fig. 5) than the ABCG2nox fraction in the Matrigel plug, 10 days after implantation. This is expected, since higher self-sufficiency would contribute to higher cell survival and resistance to differentiation of ESCs during subcutaneous implantation. We noted that a small fraction of the ABCG2 nox cells in the Matrigel plug continued to express high Nanog ( Fig. 4D; Supporting Information Fig. 5). Considering that the Matrigel plug microenvironment is hypoxic, we speculate that a few of the ABCG2nox cells in the Matrigel plugs might have undergone transient reprogramming to acquire ABCG2hox phenotype. We also noted that the ABCG2hox xenograft-derived cells did not form secondary teratomas suggesting that the ABCG2hox fraction exhibited EC-like characteristics of self-sufficiency for a transient period of time in vivo but did not exhibit the full-blown EC-like malignant phenotype of teratocarcinoma as does Tera-2. Taken together, we concluded that the ABCG2hox fraction underwent a temporary but measurable phenotypic switch to an enhanced stemness state of extreme self-sufficiency but did not exhibit malignant transformation to teratocarcinoma. HIF-2a-Mediated p53 Suppression Maintained the Enhanced Stemness State Because the pluripotency state is stabilized in a metastable state, wherein Nanog is maintained within a threshold level, the high Nanog expression state is unstable and therefore returns to a basal level within hours to days. Multiple mechanisms, including p53 activity are likely involved in maintenance of a balanced state of Nanog expression. We investigated the molecular mechanism of how the ABCG2hox fraction maintained a state of high Nanog and associated self-sufficiency for a prolonged period of time without returning to the basal state of pluripotency. We speculated that high HIF-2a might suppress p53 activity to allow the high Nanog state to transiently retain stability and tested this idea as follows. First, we investigated the phenotypic switching of ABCG2nox to ABCG2hox cells when they were made deficient in HIF-2a and HIF-1a. Both HIF-1a and HIF-2a were downregulated in BGO1 cells by siRNA gene silencing (Supporting Information Fig. 6a) and also by treatment with FM19G11 (Calbiochem), an inhibitor of HIFs. HIF-deficient BGO1 cells were then exposed to hypoxia/reoxygenation to obtain the ABCG2hox fraction. The inhibition of HIF-2a by siRNA as well as FM19G11 led to an eight-to ninefold reduction of the ABCG2hox fraction (Supporting Information Fig. 6b). In contrast, the inhibition of HIF-1a by siRNA gene silencing of HIF-1a did not affect the ABCG2hox fraction (Supporting Information Fig. 6b). These results suggest that HIF-2a and not HIF-1a was required for the survival and/or expansion of the ABCG2hox fraction during hypoxia/reoxygenation. Next, we investigated the potential HIF-2a-mediated suppression of p53 by treating the ABCG2hox fraction with the HIF-2a inhibitor FM19G11 or siRNA HIF-2a and then determining cell survival and the expressions of p53. We found that HIF-2a inhibition resulted in a fourfold increase in apoptosis as measured by caspase-3 activity (Fig. 5A) and corresponding increase in p53 expression (Fig. 5B). The enhanced stemness state Nanog and ABCG2 expressions were downregulated (Fig. 5C). These results suggested that downregulating HIF-2a led to increased expression of p53 and associated differentiation of the enhanced stemness state. We interrogated whether the increased p53 activity was associated with increased apoptosis following HIF-2a inhibition. First, the HIF-2a inhibitor-treated ABCG2hox fraction was treated with either siRNA p53 or pifithrin-a, a p53 inhibitor. We found that siRNA gene silencing of p53 enhanced survival of the ABCG2hox fraction (Fig. 5A). A similar result was obtained with pifithrin-a treatment (data not shown). These results suggested that HIF-2a was involved in the inhibition of p53-mediated apoptosis/differentiation of the ABCG2hox fraction. We concluded that high HIF-2a suppressed the p53 activity to enhance the survival of ABCG2hox cells. HIF-2a Modulates the p53/MDM2 Oscillation System Because, p53 is associated with p53/MDM2 oscillation having a period of high MDM2 and low p53 state, we evaluated the potential role of HIF-2a in altering the oscillation to sustain a high MDM2 and low p53 state in the ABCG2hox cells. First, the ABCG2hox cells were treated with Nutlin-3, an MDM2 inhibitor, and the survival as well as differentiation of the treated cells were evaluated by performing appropriate assays. We noted that Nutlin-3 inhibition led to a ninefold reduction in the ABCG2hox fraction (Supporting Information Fig. 6b). Next, we observed a four-to fivefold reduction in Nanog, Oct-4, and ABCG2 levels (p <.05; data not shown). These results suggest that a high MDM2 state was associated with the enhanced stemness state. Next, we characterized the effect of HIF-2a inhibition in the p53/MDM2 oscillation state of ABCG2hox cells by measuring the protein levels of p53 and MDM2 by InCell western and ELISA over a period of 3 weeks. The results obtained by performing InCell western are shown in Figure 6. We found a p53/MDM2 oscillation pattern in the ABCG2hox within the first 12 hours of hypoxia (Fig. 6A). The oscillation then ceased, but a high MDM2 and low p53 state persisted for 2 weeks (Fig. 6B). We found similar evidence of the p53/ MDM2 oscillation by the ELISA measurement (data not shown). Most importantly, HIF-2a inhibition by FM19G11 led to a p53/MDM2 oscillation of high p53 amplitude, and low MDM2 state (Fig. 6C), and then return of both p53 and MDM2 levels to their basal state (Fig. 6D). We noted that the p53/MDM2 oscillation in the SSEA3/ABCG2 fraction, that did not show high HIF-2a, also did not exhibit a high MDM2 state (Supporting Information Fig. 7). These results suggest that BGO1-derived ABCG2hox exhibited an abnormal state of p53/MDM2 oscillation characterized by a sustained high MDM2 state that lasted for more than 2 weeks. Treatment with HIF inhibitor FM19G11 prevented the occurrence of this sustained high MDM2 state. To further investigate the role of HIF-2a in the initiation of the abnormal high MDM2 state of the p53/MDM2 oscillation, we performed siRNA HIF-2a silencing of BGO1 before subjecting cells to hypoxia/reoxygenation as described in Sup-porting Information Figure 7. The post-hypoxia ABCG2hox was analyzed by ELISA assay to obtain p53 and MDM2 protein levels. We also performed the experiment in H9 hESCs that showed the presence of an ABCG2hox phenotype (Supporting Information Fig. 3). The result obtained with BGO1 was similar to data described in Figure 6D. The findings on H9 are described in Supporting Information Figure 7c, 7d. First, we noted that H9-derived ABCG2hox showed a similar pattern of an abnormal state of low p53 and high MDM2 that was sustained for 2 weeks. Second, siRNA HIF-2a treatment lowered the MDM2 level to a basal state in H9 (Supporting Information Fig. 7d). We concluded that the hypoxiainduced HIF-2a was associated with the transient stabilization of a high MDM2 and low p53 state in the ABCG2 hox fraction of both BGO1 and H9 hESCs. DISCUSSION hESCs are pluripotent stem cells derived from inner cell mass of developing embryos and have adapted to specialized in vitro culture conditions to maintain an undifferentiated state with self-renewal capacity (stemness state). We now report that a rare fraction of ESCs, the ABCG2hox fraction, undergoes transition under hypoxia/reoxygenation into a transient, but reasonably persistent phenotype of high HIF-2a, Nanog, Oct4, and ABCG2 activity. The phenotype was associated with an abnormal p53 and MDM2 oscillation characterized by low p53 and high MDM2 state. This phenotype was correlated with a state of enhanced stemness that was permissive for survival and self-sufficiency. Importantly, in the context of tissue repair and regeneration, the enhanced stemness state is characterized by high cytoprotective activity. Whether Thus, we propose that hESCs use a novel mechanism of enhanced stemness reprogramming to acquire a higher degree of functionality for regeneration, where HIF-2a, turned on by hypoxia and oxidative stress, sustains functional activity for at least 2 weeks after the initial induction. Although generalization of the enhanced stemness reprogramming as a defense strategy common to stem cells would be highly speculative, we suggest that if used, this potential strategy of cytoprotection (altruism) might benefit hESCs as well as adult stem cells to survive and maintain stemness and regenerative potential at the site of injury. Sites of tissue injury are subjected to cycles of hypoxia/ reoxygenation during the repair process. These sites appear to attract stem cells whose roles are still poorly understood although there is a growing understanding that stem cells may assist in the repair process either directly or through production of critical factors. Here we subjected hESCs to hypoxia/reoxygenation mimicking the site of injury. The hESC model has the advantage that distinct phenotypes can be discriminated in vitro and in vivo and the factors that maintain the stem cell state have been well elaborated. The time frame of this hESC cell phenotype of enhanced stemness state, characterized by higher degree of self-sufficiency and cytoprotective ability, corresponds to prolonged and abnormal state of p53/MDM2 oscillation. This altered state of the oscillation system was associated with a high HIF-2a activity since inhibiting HIF-2a led to phenotypic differentiation of the enhanced stemness phenotype. Thus the HIF-2a and p53 systems, coordinately upregulated during hypoxia and oxidative stress, could contribute to the emergence of a rare fraction of hESCs harboring high HIF-2a and low p53, that is, a state of enhanced stemness state of self-sufficiency. This transient state now permitted hESCs to acquire a highly cytoprotective, regenerative activity. This cytoprotective activity appears to be mediated by a high level of functional ABCG2, an efflux pump that secretes GSH, a potent endogenous antioxidant. We showed that the enhanced stemness state was associated with a high level of GSH secretion, with GSH secreted into the CM, which was capable of cytoprotecting cardiomyocytes and primitive neuronal cells represented by SH-SY5Y cell line from the toxic effect of hypoxia/reperfusion injury in vitro as well as chemotherapeutic induced BM cell cytotoxicity in vivo. Whether other factors in the CM act cooperatively with GSH remains to be determined, however, these observations strongly support the notion that stem cells in sites of injury and repair condition the environment and may contribute effectively to the regenerative process. The suppression of p53 was required to initiate and maintain the enhance stemness state, and therefore, the cytoprotective activity. However, one potential trade-off of p53 suppression would be the imbalance in the stemness state that could lead to malignant transformation, a major concern of current stem cell-based therapies. In this regard, we noted the observation that MDM2 is highly expressed in EC and positively regulates FGF-2. Although we found that the ABCG2hox cells were highly teratomagenic, ABCG2hox cells did not exhibit teratocarcinoma activity, suggesting that the enhanced stemness state did not lead to malignant transformation. One reason may be that the transient nature of the enhanced stemness state functions as a possible safety mechanism that is built into stem cells to mitigate against malignancy. In this context, it would be of crucial importance to determine how this safety mechanism is overridden, for example, by an oncogenic event, in the switch to a malignant phenotype. Our model might be ideally suited to probe this vital question. CONCLUSION In summary, we now propose a novel mechanism whereby hESCs might contribute to healing when exposed to an extreme oxidative stress state of hypoxia/reoxygenation. In this potential mechanism, a fraction of highly undifferentiated stem cells acquire a transient state of enhanced stemness reprogramming characterized by high antioxidants and their secretion, where p53 is temporarily downregulated. Thus in this model, and perhaps counterintuitive, a few stem cells can acquire a higher degree of self-sufficiency to resist differentiation in the hypoxia/oxidative stress microenvironment to protect other cells from oxidative stress damage. Is this enhanced stemness reprogramming then a form of cell altruism that could benefit the organism from the earliest developmental beginnings?

A Cache Technique for Synchronization Variables in Highly Parallel, Shared Memory Systems by Caches have traditionally been used to lower the average latency of memory access. When paired with the individual CPUs of a multiprocessor, they have the additional benefit of reducing the overall load on the processor-memory interconnection. Since synchronization variables have been identified as centers of memory contention, we have looked at methods of utilizing the cache to minimize this effect. A technique of polling the cache is proposed to deal with this problem. Consistency within the caches is maintained with a bottleneck-free update facility that exploits the topology of the multistage network. Since an indiscriminate broadcast-on-write policy can lead to severe network congestion at high levels of parallelism, we selectively invoke these updates from the software. We illustrate our methods with a number of useful synchronization algorithms and present simulation results that support the feasibility of our design. In addition to providing support for basic synchronization operations, our methodology is generally applicable to all parallel algorithms that utilize polling.

Hunter-class frigate Tender process In April 2016 the government announced a competitive evaluation process between Navantia, Fincantieri and BAE Systems for the Future Frigate Program. Additionally, a tender for the combat system was also held between Saab and Lockheed Martin. In October 2017, the government announced that the Aegis combat system and a Saab tactical interface would be used for the class. Navantia (Evolved F-100) Navantia offered an evolution of its F-100 base design, which forms the basis for the Hobart-class destroyers currently being built in Adelaide for the RAN. In 2014, the Australian Government commissioned a study to use the Hobart-class hull which Navantia claims shows it could be adapted to meet the requirements of the Future Frigate program, including integration of the CEAFAR radar and Saab 9LV combat system. Based on this study, a Navantia-designed Future Frigate would have 75 per cent systems commonality with the Hobart-class destroyers. Systems on the Hobart class include a 48-cell Mk41 vertical launch system, five-inch Mark 45 naval gun, undersea warfare capabilities including a hull mounted sonar and active and passive towed variable depth sonar, as well as the capability to operate a MH-60R "Romeo" Seahawk. The Hobart-class Air Warfare Destroyer program has attracted criticism for cost and schedule over-runs: by 2015 the program was three years behind schedule and $800 million over budget. In late 2015, Navantia was selected to bring a shipbuilding management team into government-owned shipyard ASC as part of the AWD reform program. Following the reform program, initiated by ASC prior to Navantia management integration, ASC has stated that "when we reach our budget on ship three...we will be as good as the other Aegis yards in the world." Fincantieri (Modified FREMM) Fincantieri offered the anti-submarine warfare (ASW) variant of its FREMM frigate (Bergamini class). The Italian Navy will purchase four ASW FREMMs and six of the general purpose variants. The vessels are equipped with an Italian weapons and sensor suite, including a 76 mm OTO Melara gun fore and aft, a 16-cell vertical launch system (VLS) for missiles including the MBDA Aster 15 and 30. Fincantieri says that the general hull configuration of the Bergamini design will require little or no modification to meet Australian requirements, including the incorporation of the CEAFAR radar, although it has confirmed that some redesign would be required if Australia requires the incorporation of the US Navy Mark 45 five inch naval gun. However, when discussing the FREMM, Minister for Defence Industry Christopher Pyne said that "the weaponry of course will be United States' weaponry. That will be integrated into the winning vessel." In 2016, Pyne stated that "one of the advantages for this company is that this vessel has been built, it is already in operation. One of the disadvantages is that the company doesn't operate here." BAE Systems (Type 26) BAE Systems offered an export variant of its Type 26. First conceived in 1998, the UK's Future Surface Combatant program was originally intended to replace the Royal Navy's Type 22 and Type 23 frigate fleets. In 2010 BAE Systems were given a four-year contract to fully design a new class of warship, the Type 26 or 'Global Combat Ship.' The 2010 Strategic Defence and Security Review announced that the Royal Navy would procure 13 of the vessels, however repeated delays saw the program scaled back to eight vessels with five slightly smaller, less capable but substantially cheaper vessels, that being the Type 31 warships, which have a target unit cost of just £250m off the forecourt, also being ordered by the Royal Navy for missions which do not require the level of technology and same level of survivability. Construction of the ships in the UK was expected to start in 2016, but was delayed with critics saying it was "due to the Minister of Defence's attempts to save money." Work began in 2017 with the first vessel set to enter service in the early 2020s, after Australia’s build program commences. The Royal Navy’s eight vessels are expected to cost £8 billion (AUD $13.8 billion). The Type 26 'Reference Ship Design' will be equipped with an advanced anti-submarine warfare (ASW) capability, a 24-cell strike length Mk 41 VLS for long-range strike weapons such as the Tomahawk, a 48-cell vertical launch silo (VLS) for Sea Ceptor anti-air missiles, a 5-inch gun, and is capable of landing a Chinook helicopter on its flight deck. In April 2018 BAE claimed it would be able to offer the best ASW performance of the three contenders, despite not having a ship of the class in the water at the time. Design and construction The Hunter-class frigate will be an Australian variation of the Type 26 class frigate that is to be operated by the Royal Navy from the mid-2020s. The class will have a 8,800-tonne (8,700-long-ton; 9,700-short-ton) full load displacement and will be approximately 150 metres (490 ft) in length. The vessel will be capable of sailing in excess of 27 knots (50 km/h; 31 mph) and will have a full complement of 180 crew. A Saab tactical interface with the Aegis combat interface will be used. The vessel will be able to carry one MH-60R ASW helicopter, and has the ability to host other Australian aircraft such as the MRH90 helicopter. The ships will be built by BAE Systems Australia at the ASC's Osborne, South Australia shipyard.

Digitization and sustainability of local collection : an observation of digitization activities among Malaysian universities libraries Malaysian academic libraries have responded well to the governments IT agenda. They are among the first group of libraries to embark on digitization projects. This paper attempts to discuss the types of materials digitized, guidelines, policies and provision for access to the digitized materials. It also discusses some of the issues and challenges in this process of enriching the local digital content. An appropriate training module with practical guidelines and policy is recommended to ensure continuity and sustainability of these projects.

Web Application for Ration Distribution System This paper proposes the advanced Ration Distribution System, named as Web Application for Ration Distribution Management. Ration card is extremely vital for every home. It performs as address evidence for various purposes. It is used for various fields such as for obtaining family members details, to obtain gas connection, etc. Every home has a ration card to obtain the various materials (sugar, rice, oil, kerosene, etc.) from the ration shops. But if that ration card will be lost then owner of that card need to make new one and that takes a lot of time instead of that we can do all the things by using website and plus point is using this web application we can do the online secure transactions as well. This will help to make digitalization process and helps to reduce the corruption.

Applying Data Mining Techniques to Understand the Impact of Information Technology on Organizational Productivity We explore conditions under which investments in Information Technology (IT) have a statistically significant impact on productivity. Rather than using a traditional analytical technique such as regression, we use two data mining techniques (i.e. regression trees and regression splines) for doing data analysis and integrated their results. Our analysis indicates the following: 1) IT investments have a positive impact on organizational productivity only when IT investments meet certain conditions; and 2) the IT impact is not uniform but varies depending on the amounts invested in other related areas, such as Non-IT Labor, Non-IT Capital, and IT Stock. Thus, our study leads to some suggestions to top managers that organizations should assess the current state of investments in Non-IT Labor, Non-IT Capital, and IT Stock before making any further commitments to invest in IT as this current state partially determines the potential impact of additional investments in IT on organizational productivity.

Well, the car door worked fine tonight. I was fine till I got to the arena, plugged in my laptop, reached for my glasses — and they were gone. Actually, they’re at home. Now, I can see the game just fine. But I cannot see this screen or the stats or any other printed words smaller than about 18-point type without the aide of my specs. So, I got up and started out to return home to get my glasses … and Deb Ritschel, GM of the Civic Center, offered me the use of her reading glasses. Bless her heart. So here I sit, hoping the frames don’t break from the stress of my giant head. But I can see what I’m writing. I look a little strange in tiny glasses designed for a woman, but they are functional, and if I cared how I looked I would have ponied up for plastic surgery decades ago. Braves are back on the court for the official pregame warmup. The band is playing. A dozen kids already are lined up along the sideline, watching the warmups, slapping a few palms with players and staking out position for the lineup lap. The refs tonight are Mike Stuart, Mark Whitehead and Kelly Self. Big game for BU. A win tonight is a nice launch into the two-game road trip to Iowa this weekend. A loss means an 0-3 start, with a real danger of being 0-5 in a week. My colleague Dave Reynolds predicts a five-point Bradley win. I’ll go with that — not because he’s been on top of it (he’s but 8-5 this season), but because I think he’s right this time. By the way, for those of you who discuss the program on the bradleyhoops.com forum: We’re transferring servers, and there’ve been some operational issues the past few days. At this point, you still might be having trouble accessing the forum through the link on the bradleyhoops site, or through your bookmark. Hopefully our people will get that fixed soon. Meanwhile, you can still access the site through the pjstar.com forums. Here’s a link. During the lineups tonight there are about 60 kids lined up along the sidelines and baselines to slap hands with the BU starters. Nice tradition that started quite a few years ago. It caught the attention of the Stadium Journey visitor last month. The SJ site posted this review of the Bradley experience at Carver. A little ugly play on both ends.

Having decided not to buy a car because of the expense of operating and maintaining the thing, I was slightly distressed to hear that my bike needed to go into the shop for work. Just as with a car, I have absolutely no idea whether it actually needed the front hub realignment they gave me along with the replaced spoke I came in for. The nice thing is, though, that at worst my ignorance cost me $15, rather than the $1400 I was taken for the last time I went to see a mechanic.

New Delhi, Apr 12 (ANI): Union Minister for Minority Affairs Mukhtar Abbas Naqvi spoke to mediapersons after Bharatiya Janata Party (BJP) delegation met Election Commission of India. Naqvi said, "Rahul Gandhi has crossed all limits by using such level of unparliamentarily language for the Prime Minister. He's chief of Congress's 'gaali gang'. We have demanded action be taken against his statements, which aren't based on facts." BJP delegations have met ECI over the petition in Supreme Court for criminal contempt against Congress president Rahul Gandhi. BJP alleged that Rahul Gandhi made 'fake' comments over Prime Minister Narendra Modi.

Seasonal Poverty and Seasonal Migration in Asia Four in five poor people in the Asia and Pacific region live in rural areas. Crop cycles in agrarian areas create periods of seasonal deprivation, or preharvest lean seasons, when work is scarce and skipped meals become frequent. In this paper, we document this phenomenon of seasonal poverty and discuss existing formal mechanisms for coping with it. We then focus on seasonal migration from rural to urban areas as a potential coping strategy and review the evidence on the effects of encouraging seasonal migration through transport subsidies. Over the past 10 years, we have conducted a series of randomized control trials in Bangladesh and Indonesia that provided rural agricultural workers with small migration subsidies to pay for the cost of round-trip travel to nearby areas in search of work. This paper summarizes the lessons learned from this multicountry, multiyear series of seasonal migration trials, the implications of these results for spatial misallocation, urbanization, and growth, and the replicability and relevance of this and other policies encouraging domestic migration more broadly for other areas in the Asia and Pacific region. In the Asia and Pacific region, poverty is highly concentrated in rural areas. Four out of every five poor people in Asia live in rural areas (Asian Development Bank 2007), and the rural concentration of poverty is even starker within some countries. In Viet Nam and Cambodia, for instance, rural areas account for 90% of all poor people (Balisacan, Edillon, and Piza 2005). In Bangladesh, 35% of rural households are poor, compared to 21% in urban areas. In Pakistan, these figures are 36% and 18%, respectively, and in the Lao People's Democratic Republic, 29% of rural households are poor, almost three times the share for urban residents (10%) (World Bank 2018). 1 As a subregion, South Asia is the most rural in the world and the only one besides sub-Saharan Africa in which the share of the population living in rural areas still exceeds that for urban areas (Figure 1). A majority of the population in rural areas, coupled with a concentration of poverty in these same places, translates into a large number of poor rural households in relative and absolute terms. Most of these households are engaged in agriculture and are vulnerable to more severe and frequent shocks compared to urban households. In this paper, we discuss the seasonal variation in poverty within rural, agrarian areas and summarize findings from 10 years of research on a strategy many poor rural households use as a way to cope with seasonal deprivation: temporary, within-country seasonal migration. Both pilot and larger-scale interventions conducted in Bangladesh and Indonesia reveal that policies that encourage seasonal migration by providing transport subsidies and lowering migration costs can, under certain conditions, help poor families mitigate the adverse effects of seasonal poverty by expanding their access to urban labor markets. A. Internal Migration Internal migration-permanent, temporary, seasonal, or cyclical-is a common coping strategy among rural households and has played a key role in accelerating urbanization in the region. There are 282 million internal migrants in Asia, which account for over one-third of all internal migrants globally (UN DESA 2013). 2 In the region, a lot of internal migration has historically been of a more permanent nature-such as relocation to urban or manufacturing areas-but this pattern has slowed down in several countries since the late 1990s. In Indonesia, Malaysia, and Viet Nam, this deceleration is likely related to an aging population (who are less likely to migrate), economic growth, and a reduction in interregional wage gaps. The People's Republic of China (PRC) is an exception to this trend, as within-province migration actually increased by well over 100% between 1990 and 2000, likely due to the loosening of restrictions on migration to its large cities (Bell and Charles-Edwards 2014). In contrast to the deceleration in permanent migration, temporary, seasonal, and circular migration have increased in Asia over the last few decades as temporary employment opportunities in urban and manufacturing centers expanded. Given crowding and limited access to housing in urban areas, a lot of the recent rural-urban movement has been temporary and reversible in nature (Deshingkar 2006). Beyond the draw of better employment opportunities in cities, there are also a couple of push factors that drive rural people into internal, seasonal migration: predictable preharvest lean periods in agricultural crop cycles during which labor demand is low in rural areas, and high levels of vulnerability to unanticipated shocks, which further exacerbate income fluctuations. B. Risk and Vulnerability in Rural Areas as Drivers of Internal Migration The 2014 World Development Report on risk and opportunity analyzed the incidence of adverse shocks using household survey responses and found a much higher prevalence of shocks in rural areas compared to urban ones (World Bank 2013). In India, 62% of rural households reported experiencing at least one negative shock within the previous 12 months and 24% reported two or more. In the Lao People's Democratic Republic, 36% of rural residents reported a negative shock compared to 12% of urban residents. Economic activity in rural areas is much more weather dependent and less diverse than in urban areas, and rural residents report facing both aggregate shocks such as natural disasters (e.g., droughts and floods) and idiosyncratic health shocks. While health shocks are also common in urban areas, natural disasters do not appear to be as big a concern for urban households when compared to rural ones (World Bank 2013). Acute shocks in rural areas such as droughts and floods certainly contribute to the heavy flux of temporary migration observed in Asia. In 2010, for instance, 14 million people were temporarily displaced in Pakistan because of floods (Lucas 2015). And in 2001, almost two-thirds of all people in Bolangir, a district in the Indian state of Odisha, migrated during a drought (Deshingkar 2006). It is estimated that between 2011 and 2050, as many as 26 million people in Bangladesh may have to migrate because of floods, storms, riverbank erosion, and sea-level rise (). Alongside these dramatic responses, a lot of temporary migration is recurring in nature, as people move not only because of unanticipated shocks but also to cope with predictable seasonal variation in employment opportunities in rural areas. In northern Bangladesh, every year, one-third of poor households in rural areas send a migrant to work elsewhere in the country for an average of 2-3 months (Khandker and Mahmud 2012). This massive movement of people that occurs at a predictable time of the year is the result of seasonal fluctuations in income and work opportunities in areas of origin due to the agricultural crop cycle and not because of unexpected natural disasters. In areas with little crop diversification and distinct cycles, there is a lean season between planting and harvest periods, a time when agricultural jobs are scarce, harvest income is yet to come in, and poor households regularly skip meals and suppress expenditures. Though food insecurity is a nonnegligible issue for poor households throughout the year, there is a stark increase in extreme levels of deprivation during this period. In 2006 (a typical year in terms of agriculture), 47% of poor households in northern Bangladesh experienced hunger during the agricultural lean season (Khandker and Mahmud 2012). In contrast, only 9% of households went hungry outside the lean season that same year. More recent data we have collected in the region show spikes in hunger in two different lean periods within the course of a year: from August to October and briefly around March (Figure 2). The main rice crop (aman) is typically harvested from November to January, and the secondary rice harvest (boro) is in April. The periods before each of these harvests are marked by heightened food insecurity among landless rural households: whereas over half of poor households reported at least sometimes skipping meals during the main lean season (from August to October), fewer than 25% reported doing the same once the aman harvest was underway or after the boro harvest. C. The Effects of Seasonal Poverty Income fluctuations, even when seasonal and expected, can have dramatic consequences for the poor who live close to subsistence and are unable to accumulate savings and smooth consumption. In rural areas in the PRC, for instance, households in the bottom 10th percentile in wealth respond to a drop of 100 yuan (CNY) in income with a decrease in food and nonfood expenditure of CNY40 on average. In contrast, for the top-third of households considered "nonpoor," the same drop in income results in just a CNY10 decrease in expenditures (World Bank 2013). Such consumption drops among the poor have serious implications for health and well-being, especially for pregnant women and young children. Skipping meals and decreasing food intake leads to stunting, nutrient deficiencies, and other health issues, with potential repercussions for future cognitive capacity and earnings. Given the concentration of poverty-and vulnerability to agricultural cycles-in rural areas, it is perhaps not surprising then that Bangladesh has the fourth-highest prevalence of stunting among poor households around the world, and South Asia remains the region with the largest number of stunted children under the age of 5 (World Bank 2018). Increases in population per hectare of crop land in several Asian countries forebodes further competition for the few tasks that are available in rural agrarian areas during the preharvest period. As is, there is a danger of potentially worsening health outcomes for the rural poor over the next few decades (Deshingkar 2006). D. Seasonal Migration as a Policy Tool to Counter Seasonal Poverty The limited set of tools readily used by the rural poor to cope with seasonal income fluctuations include starting a nonagricultural business, engaging in nonfarm employment, or temporarily migrating. Business creation and nonfarm employment are strategies most commonly used in sub-Saharan Africa (World Bank 2012) and can help households diversify away from agricultural income and its cycles. At the same time, these are reliable coping tools only if there are profitable opportunities within rural areas, which appear not to be the case in large parts of rural Asia (World Bank 2012). Social or cultural constraints may further restrict households' ability to engage in nonfarm employment. Poor rural households in Bangladesh, where women are less likely to work outside the home due to cultural norms, have even less diversified income sources than in neighboring India, for example (World Bank 2013). In contrast to rural-based livelihood diversification strategies, seasonal migration may be profitable even when local rural areas are poor and offer sparse employment opportunities. If travel between villages of origin in rural areas and urban destinations is manageable and not too expensive, then spatial wage gaps may create an arbitrage opportunity. Migrants can take advantage of employment opportunities in urban areas and send remittances or bring money back home at the end of their period of employment to help their family cope during the lean period. One study estimates that migrants in Dhaka send as much as 60% of their income back home (Deshingkar 2006), while another shows that a quarter of rural Indonesian households have a migrant, with 85% of urban migrants sending money home (Lu 2013). Benefits from seasonal migration and its role in smoothing consumption may extend beyond averted hunger alone and into other gains in human capital. In rural Bangladesh, school attendance is higher among children in households with temporary migrants than in those without; and in Bangladesh, India, and Nepal, households with seasonal migrants report higher expenditures on education than nonmigrant households (). Our research, as described in detail in section III, employs a series of randomized control trials (RCT) to explore whether encouraging more seasonal migration from rural areas helps poor households mitigate the adverse effects of seasonal deprivation. Since 2008, we have been testing the effects of small migration subsidies ($8.5-$19) in rural Bangladesh and have found that this low-cost intervention increases consumption and income among treated households relative to a control group. The main economic results are reported in studies by Bryan, Chowdhury, and Mobarak (BCM) ; Akram, Chowdhury, and Mobarak (ACM) ; and Lagakos, Mobarak, and Waugh (LMW). BCM and ACM show that the impact of a one-time subsidy also carries over to subsequent years, as treated households are more likely to send a migrant up to 3 years after the initial treatment. However, LMW show that the financial benefits of migration are tempered by the disutility associated with bad living conditions experienced in destination areas, particularly urban slums. We are continuing to develop further evidence on this program with each new year of implementation and asking new questions as the program scales. Over the last 10 years, through multiple phases of testing this intervention, we gradually moved from the original RCT covering 1,900 households to testing a program called "No Lean Season," which was run by a separate entity and disbursed zero-interest migration loans to over 80,000 households in the 2018-2019 lean season alone. 3 This scale-up required more local engagement and resources and adjustments to the delivery system, moving from an intervention carefully monitored by researchers at each step of the way to one with greater autonomy for implementing partners and a slightly more hands-off approach to test scalability. At the same time, the shift from RCT to the program demanded attention to effects that might not be relevant to an initial study but that come into play as the target population and area expands. With this transition as motivation, ACM focused on general equilibrium effects on nonmigrant households in villages of origin, considering the effects of offering migration subsidies to some households on the consumption levels and incomes of other poor households in the same village. Other papers have explored the noneconomic effects of encouraging migration, such as those on beliefs, attitudes, and social norms (Mobarak, Reimo, and Thachil 2018), effects on intimate partner violence (Mobarak and Ramos 2018), pressures toward urbanization in the long run (Chowdhury, Mobarak, and Reimo 2018), effects on informal insurance networks (), and the relevance of the intervention to other settings, such as in rural eastern Indonesia (). We are also preparing to study (i) whether seasonally timed consumption loans (as opposed to seasonal migration subsidies) is the right policy response in a different setting, with a new experimental design in Nepal; (ii) the effects of the scaled-up version of the program on urban labor markets in Bangladesh; and (iii) the role of implementation changes on targeting and migration outcomes. 3 "No Lean Season" was a program run by the Beta division of Evidence Action and implemented in Bangladesh through its partner RDRS Bangladesh. One of the authors of this document was a postdoctoral fellow with Evidence Action, and the entire research team engaged with Evidence Action on a regular basis to provide input for program decisions. No Lean Season, as funded by Evidence Action, was discontinued after the 2018 lean season, but RDRS Bangladesh has continued to offer migration loans. The research team has not been involved in these subsequent activities. E. Other Policy Options to Counter Seasonal Poverty Explicit support for seasonal migration through transportation subsidies is only one option in a suite of potential policies to address seasonal deprivation in rural areas. In Japan; the Republic of Korea; and Taipei,China, for example, increased crop diversity-which makes seasonal variations in agricultural employment opportunities less acute-and investments in transportation infrastructure to improve access to other regions played key roles in the national development trajectory (UNFPA 2016). With improved and expanded transport networks, rural households could both commute and temporarily migrate to cities, better allocating their labor between local agriculture and other employment options-between members as well as over time. On a related note, Asher and Novosad show that a rural road construction program in India allowed connected villages to reduce their reliance on agricultural income earned inside the village. Likewise, Bangladesh (and other countries in South Asia and Southeast Asia) could benefit from investments in deeper and safer transportation networks and in better housing options for the poor in urban areas. This strategic funding could transform temporary migration from a coping strategy to a predictable and desirable yearly pattern for rural residents, while enabling local governments and employers to better manage and respond to the influx. Improved transportation can also expand commuting opportunities, which would counter people's need to migrate and live in destination areas for long periods. However, for the foreseeable future (until transportation networks vastly improve or rural residents have better access to local jobs year round, or both), explicit support for seasonal migration will likely continue to play a role in helping the poor cope with seasonal deprivation. In the next section, we discuss a broader range of policies that governments and development organizations typically employ to help the rural poor deal with employment and income shocks, including guaranteed work schemes, universal basic income, crop insurance, and microfinance, and we point to the gap that can be filled by seasonal migration subsidies. In section III, we discuss in greater depth the design and results of the seasonal migration support programs we have implemented and tested over the last 10 years. We discuss the implications of our research for other countries in the Asia and Pacific region in section IV and subsequently conclude. II. Rural Development Policies to Address Seasonal Poverty One of the largest and best known seasonal income support programs in the world is India's National Rural Employment Guarantee Act (NREGA). Under this program, all rural workers in the country are in theory entitled to 100 days of guaranteed employment per year and are primarily hired for projects that support local community development, such as irrigation and road construction. Other common rural poverty programs include promoting financial services such as microfinance (a concept that also originated in South Asia), crop insurance, and support for savings (Table 1). Many governments around the world have also experimented with cash transfers-either unconditional or conditional on a specified activity like school attendance-and transfers in the form of food subsidies or free food distribution have been deployed in acute situations around the world for decades. Besides these formal policies, poor households have long used informal coping systems as well. In times of distress, they rely on their social networks for gifts and transfers, on informal loan providers, on temporary or permanent migration, or on invoking the very costly strategy of cutting back on consumption, or a combination of these. In this section, we provide an overview of commonly employed antipoverty policies for rural areas and discuss the evidence of impact of such programs, drawing on data from Asian countries whenever possible. We close the section by explaining how subsidies for seasonal migration can fill a gap left by these programs. A. Guaranteed Work Schemes NREGA, enacted in 2006, is an ambitious program to address rural seasonal poverty in India. It is a cash-for-work program that provides guaranteed employment during the lean season for rural households across the country. Within 6 years of inception, this program had provided over 12 billion person-days of employment for the rural poor. In the 2017-2018 fiscal year, almost 80 million individuals worked under the scheme (NREGA 2018). NREGA relies on "self-targeting," in that offered wages are deliberately low relative to the market, to ensure that only those facing weak employment opportunities will seek out the program. Despite this, no state has been able to provide all of the employment that rural workers have demanded and are entitled to (World Bank 2012, ). In fact, in the 2017-2018 fiscal year, the average number of days worked per participant household was 46 days, well below the guarantee of 100 days per person (NREGA 2018). Nevertheless, there is evidence that the opportunities offered by NREGA have had a spillover effect on the private sector, pushing up market wages and benefiting the poor more broadly (Imbert and Papp 2015;Muralidharan, Niehaus, and Sukhtankar 2018). Studies report other positive effects of NREGA, including on consumption, assets, nutrition, education expenditures, and women's empowerment (Das and Singh 2013, Dasgupta 2017, Deininger and Liu 2013, but since the program rollout was not designed with research in mind, it has been difficult to establish some of the evidence very rigorously. In contrast, a similar rural employment guarantee scheme in Malawi was evaluated using an RCT, and it found no meaningful benefits in terms of agricultural investment, food security, or labor market opportunities for beneficiaries (Beegle, Galasso, and Goldberg 2017). This study even reported a negative spillover effect on untreated households operating in the same labor markets as the beneficiaries of the cash-for-work program. Guaranteed work schemes, even when they are effective, are very costly to operate because they must force job creation in a rural area where the structure of the agricultural economy is such that it is difficult to generate employment. This is likely related to crop cycles. Lean seasons often appear in agrarian areas during the period between planting and harvest, where cultivators must patiently wait for the crop to grow. There is relatively little to do on the farm, as weeding and other land management tasks do not require as many workers as during planting and harvest. These preharvest lean periods constitute expected seasonal downturns marked by job scarcity and are known as "monga" in Bangladesh, "musim paceklik" in Indonesia, or simply "hungry seasons" in Malawi, Mozambique, Senegal, and Zambia. The private sector does not generate sufficient employment during this period in rural areas, but it is not clear whether this is due to a market failure or is the natural result of economic conditions given the concentration of poor people with little buying power during the lean season. Nevertheless, government-led guaranteed employment programs expend vast amounts of resources to go against this tide and create local jobs each year during the lean season. Even setting aside their debatable potential as efficiency-enhancing programs that address a market failure, and instead looking at them as a social safety net for the extreme poor, food-and cash-for-work programs are difficult to design to meet local needs. The wages offered through the program need to achieve a balance between being low enough to properly target the poor and needy who should self-select into the program, but high enough to actually address seasonal poverty and be of use to those poor households (who may also face higher prices during the lean season). NREGA participants in India receive an average of 175 rupees ($2.50) per day, and there are indications that the program is well targeted for the poor (Imbert and Papp 2019). In contrast, the wages offered in the Malawi government program are apparently so low relative to outside options that only 50% of households offered employment through the system actually accepted (Beegle, Galasso, and Goldberg 2017). A job guarantee program in Bangladesh called Food for Assets (FFA) is also reportedly well targeted to the poorest households, because the nature of the work is physically demanding and the program offers low pay. Of FFA participants, 72% come from the lowest decile of the income distribution (). Job availability also generally overlaps-though not perfectly-with the lean season, as most of the work is provided between December and May. There is some evidence, however, that this program crowds out other income, as participants are required to work full days, thereby foregoing labor opportunities in private markets. As such, every 100 taka (Tk) paid through the FFA program increases household income by an average of only Tk32 (). Rural workfare programs also discourage labor migration to urban areas, as one must be present in the rural area to take advantage of the job guarantee scheme. Since these programs are more common during the lean season (both to address seasonality and because construction projects are challenging to implement during monsoon periods), they also directly compete with seasonal migration-based coping strategies that those households may have otherwise employed. Research in India reveals that districts that were early implementers of NREGA experienced a 50% decrease in seasonal out-migration relative to a comparison group of districts where the program was not yet implemented at the time (Imbert and Papp 2019). In our own research in Bangladesh, we find that displacement of migration limits the positive effect of work programs on household welfare. Simulations reported in LMW show that introducing a rural workfare program in northern Bangladesh would decrease the seasonal emigration rate of the poorest quintile by 4-6 percentage points and produce lower household welfare gains, on average, relative to migration subsidies or an alternate program providing unconditional cash transfers without imposing a requirement to remain in the rural area (LMW 2018). B. Unconditional Transfers Unconditional cash transfers (UCTs) have gained popularity in recent years as a benchmark against which to measure the impact and cost-effectiveness of other poverty alleviation programs. In LMW, we directly compare the effects of seasonal migration subsidies on household welfare to the welfare that would be generated under an untargeted UCT program. The overall welfare gain across the population produced by an untargeted UCT is by definition greater than that of a transfer conditional on migration, because a migration conditionality imposes a constraint on people, and constraints can only make rational people (weakly) worse off. However, the migration transfer generates some targeting benefits relative to the UCT, since it tends to induce only the people who actually need the support to participate, as opposed to the universal coverage of the UCT, which ends up benefiting many people who are not as poor or who did not face an acute adverse seasonal shock during the year. Therefore, when we conduct a budget-neutral comparison between migration subsidies and UCTs, we see that the migration subsidies improve the welfare of the poorest quintile by about 14% more than a UCT program (LMW 2018). These results highlight a feature of UCTs that is both their strength and their weakness: the lack of any conditionality means that anyone can receive a UCT, which includes both the nonpoor (who are not intended beneficiaries of pro-poor programs) and households who may be so poor and so constrained that they may have difficulty complying with the conditions required by a conditional cash transfer (CCT) program. A study by Baird, McIntosh, and zler highlights this particular benefit of UCTs by directly comparing a UCT program against a CCT in which transfers for adolescent girls are conditioned on school attendance. This RCT-based study shows that while the CCT produces larger effects on the schooling outcomes on which transfers are explicitly conditioned, it does not decrease marriage and pregnancy rates among young girls by as much as the UCT. This is because there is a subgroup of extremely vulnerable households who cannot comply with the conditionality and therefore do not receive benefits under the CCT program. These types of households are still helped in the UCT. This logic implies that, relative to seasonal migration support programs or rural workfare programs, a basic income transfer might better reach households that may not have a working member who is able to migrate or even work in the rural area. A substantial part of the transfers, however, would also go to those who may not be in as dire a situation as the target population. Of relevance to this difference in coverage, we also implemented an RCT in West Timor, Indonesia, designed to empirically compare the relative effects of UCTs and migration transfers directly (). Not surprisingly, take-up among eligible households is over 90% in the UCT arm, compared to 52%-59% in the CCT arm. The conditional transfer increases migration by about 30 percentage points relative to the UCT, and average gains in household income are larger for the CCT group compared to the UCT. Nevertheless, the difference in take-up rates is composed of both households that are too wealthy to be interested in migrating and households that may have been unable to migrate or find a job in destination areas. A CCT such as a seasonal migration support program cannot directly reach the latter group, a disadvantage relative to UCT programs or other unconditional safety nets. Seasonal migration-and programs supporting it-is not and cannot be a substitute for basic income, particularly for households that do not have a working member. It is also important to acknowledge that cash and food transfers for the poorest households are relevant in several circumstances besides seasonal drops in income. That said, these programs are relatively costly to implement and also vulnerable to elite capture. Ahmed et al. evaluate transfer programs targeting the ultra poor in Bangladesh, including Income Generation for Vulnerable Group Development (IGVGD), a food transfer and credit program, and Food Security Vulnerable Group Development (FSGVD), a combined food and cash transfer program. The study finds that 10% of IGVGD-beneficiary households were actually in the top three deciles of the income distribution. Only 43% and 38% of beneficiaries for IGVGD and FSGVD, respectively, were in the bottom decile; in contrast, the respective share was 72% for the FFA guaranteed workfare program (). On the other hand, the same study also reports that the food-for-work program is the most expensive way to increase household consumption, with the FFA requiring Tk440 per beneficiary per month to increase daily consumption per capita by 100 calories. IGVGD requires Tk249, while FSGVD, the combined food and cash transfer program, requires Tk156 per month. The workfare program is likely even more costly in ways that are not measured, since participation in workfare generally requires beneficiaries to forego other labor market opportunities. The coverage of these transfer programs is also precarious, as only 6%-7% of the Bangladeshi poor are actually covered by any food or cash transfer or workfare program (), a statistic likely related to the high cost of implementing such schemes. These programs are therefore inadequate in their present form to address the recurrent seasonal poverty that afflicts well over one-third of the rural population in northern Bangladesh each year. Seasonal migration support may therefore serve as a useful policy complement to these existing social safety net programs in this setting-and similar ones with low safety net coverage and high seasonal vulnerability. C. Agricultural Insurance Agricultural insurance programs are designed to mitigate losses associated with extreme events and unexpected weather shocks, rather than predictable and recurrent seasonality. Index-based insurance programs circumvent the adverse selection and moral hazard problems that undermine traditional crop insurance by tying payouts to verifiable weather events or other aggregate outcomes such as average losses at the regional level. By design, then, agricultural insurance is not meant to address seasonal deprivation that arises on a cyclical basis through the regular agricultural calendar, as payouts are made based on deviations from expected patterns. A more sophisticated tool ties agricultural insurance to a safety net, as is done in Mongolia. Under this particular program, Mongolian herders fully absorb losses of up to 6% (indexed on the average mortality of adult livestock in each county); losses between 6% and 30% are covered by index insurance; and participants are automatically enrolled in a safety net program if losses surpass 30% (World Bank 2013). Though crop (livestock) insurance is traditionally aimed at landowners (livestock owners), payouts are not tied to individual output, so it is possible to expand the target population for crop (livestock) insurance from landed to landless (livestock-less) households, who are also vulnerable to extreme shocks through decreased farm (herding) employment opportunities. In fact, in an RCT in India where landless laborers were offered rainfall insurance, their take-up rate was only 4% lower than that of landowning households (Mobarak and Rosenzweig 2013). However, selling crop or weather insurance to the landless poor necessarily requires the use of index insurance, where payments are not tied to individual outcomes but to a measurement along the lines of a weather index or an aggregate outcome. This in turn implies that the policy can only insure against aggregate risks (such as weather, pest, or price shocks) and not idiosyncratic risks that poor rural households may face, such as consumption drops due to a mortality or morbidity event in the household. Empirical research on agricultural insurance also reveals that take-up is relatively low in some contexts even at actuarially fair prices, and its use has not necessarily led to higher levels of investment and technology adoption (Lybbert and Carter 2015;Carter, Cheng, and Sarris 2016). In a study offering insurance to landless households, roughly 40% of all households purchased insurance-a fairly low rate considering that subsidies of 0%, 10%, 50%, or 75% on the insurance product were randomized across households (Mobarak and Rosenzweig 2013). A new tool combining index insurance and loans, in the form of emergency loans that are made available to farmers in the event of flooding, has been recently designed and implemented in Bangladesh. The key innovation of this intervention is that, unlike other insurance forms, it does not require any upfront payment by farmers; only eligibility is determined before the crop cycle. Preliminary findings are generally positive, indicating that these emergency loans not only provide relief as expected in the case of floods, but also lead prequalified farmers to plant 15% more rice once they know they will be eligible for the loan in the event of major flooding (Lane 2018). This can generate spillover benefits for the landless poor in terms of greater labor market opportunities when the weather cooperates, particularly during the planting period (Mobarak and Rosenzweig 2013), but still cannot fully address the regular decrease in employment opportunities during the lean season. D. Microcredit and Savings Microcredit and savings consist of a broad array of programs that provide small loans for the creation or expansion of nonfarm enterprises, credit for agricultural inputs, programs encouraging savings for farm or nonfarm businesses, and any combination of these. Small loans for nonfarm enterprises became a hugely popular idea in international development in the 1990s, and South Asia is the birthplace of large microfinance institutions, including Grameen Bank and BRAC. But recent meta-analyses of RCTs on microfinance generally find modest impacts at best from this type of intervention. A review of six microfinance studies, two of which were implemented in Asia (India and Mongolia), notes that take-up rates of microloans are relatively low and the impact on family income or consumption was not statistically significant in any of the six settings (Banerjee, Karlan, and Zinman 2015). Limited take-up is to be expected even before considering loan characteristics, as not all individuals aspire to be entrepreneurs or have a promising business idea, but half of the studies also found no impact on the ownership, start, or closure of a business, while the other half only found effects in one of these outcomes. The evaluation in India found significant increases in expenditures on durable goods but not on consumption. And the evaluation in Mongolia shows positive impacts on household food consumption but not income for a group liability loan, while individual liability loans generally had no impact across the board (). A Bayesian hierarchical analysis of an overlapping set of seven microfinance studies (Meager 2019) echoes the limited power of microfinance, showing that its impact on consumption is at best small and unlikely to be transformative, with relatively little heterogeneity across beneficiaries. Notably, this study also shows that, for households with no previous business experience, as would likely be the case for most primarily agricultural households in rural areas, microfinance also has zero impact on household profits. It is clear, then, that microfinance for entrepreneurship, though potentially beneficial to some households-mainly those with existing enterprises-cannot reliably serve as an avenue for income diversification or consumption smoothing for a large number of poor agricultural households. The seasonal migration support program we have evaluated in Bangladesh is in essence a version of a microcredit program with some crucial differences from how microfinance has traditionally been conceptualized. 4 First, these loans are explicitly conditioned on migration rather than a business plan or business ownership. Second, the loans are meant to encourage job search rather than require beneficiaries to set up a business. This difference expands the relevance of the loan and removes undue pressure toward business creation, as there are likely many more "employees" than "entrepreneurs" in the world; many more would prefer a job with a stable income rather than take on the risk of starting a business. Third, our seasonal migration support was in some cases a grant rather than a loan, and in others, a loan with zero interest rate. While microfinance institutions require profitability to be sustainable, our intervention is designed to subsidize migration for the poorest households rather than turn a profit. Fourth, the repayment period for our loans has been set at the end of the lean season, typically 3-4 months after loans are disbursed. In contrast, microcredit programs typically require biweekly repayment or other short intervals (Field, Holland, and Pande 2014), which may in itself distort household decisions away from seasonal migration. In fact, data from Bangladesh indicate that clients of microfinance organizations are less likely to migrate than nonclients (Khandker and Mahmud 2012). Agricultural loan programs tailored to rural areas (to encourage investment in agricultural inputs) also move away from short repayment periods and instead offer credit at the start of the planting season and collect repayment at harvest. However, take-up of such loans is also low: a study in Mali found that just 22% of women who were offered an agricultural loan accepted it, and only the more productive farmers self-selected into taking out these loans (). Savings programs, including agricultural savings accounts, have been studied more extensively in Africa. An RCT in Malawi that encouraged savings after harvest to invest in inputs next season () found that, on average, only one-quarter of the original deposit was still in a savings account 2 weeks later. When an Abdul Latif Jameel Poverty Action Lab research team tried introducing new savings products through banks in Chile, Malawi, and Uganda, the programs generally failed, presumably because banks found it too costly to administer products in which poor people deposited very small amounts of savings each month. 5 Our data from Bangladesh indicate that the landless poor, who are most vulnerable to seasonal hunger, typically do not have anything to save, especially during the lean season. Both ACM and BCM report significant treatment effects on income and consumption from the migration subsidies introduced, but the studies do not observe any increase in savings in any of the multiple years in which the program was evaluated. The marginal propensity to consume extra income during the lean season is very high for the landless poor. The evidence suggests that it will likely be difficult to induce savings among the poor as a way to address seasonal deprivation. Fink, Jack, and Masiye identify a more promising option: a well-timed loan during the lean season can reduce farmers' desperation to suboptimally supply labor to other farms at low wages to address their short-run cash needs for meeting their family's subsistence requirements. In this treatment, loans of either cash or grains are not envisioned as encouraging leaps in productivity or business creation; rather, they are disbursed to farmers during the lean season to discourage the use of more costly coping mechanisms, particularly working in other farms. The authors find that these loans, which are also to be repaid after the lean season (effectively at a 4%-5% interest rate), do in fact encourage subsistence farmers to work on their own land during the lean season rather than in other farms, with positive repercussions for their subsequent harvest. Landless rural households may also benefit from seasonally timed consumption loans, disbursed at the beginning of the lean season and repaid at the end with no conditionality. This may be a particularly sensible solution in countries where migration already occurs at high rates and usually over greater distances and for longer periods, but where seasonal poverty remains. Our initial exploration in Nepal indicates that seasonal consumption loans may be relevant there and calls for further research on this type of consumption loan, with a closer look at repayment rates. In Zambia, repayment rates on lean-season loans dropped between the first and second years of treatment even though the harvested output increased for treated households (Fink, Jack, and Masiye 2018). Ensuring high repayment rates year after year is critical for the sustainability of such a program in the long run, and future experiments will be useful for illuminating that path. E. Temporary Migration In addition to the formal markets and mechanisms described above (employment guarantee schemes, transfers, insurance, credit, and savings programs), the poor use a variety of informal tools to cope with seasonal volatility. They may draw on support from friends and relatives, secure loans from informal money lenders, or attempt to diversify their income by engaging in informal business enterprises or by entering the labor market outside the village-or a combination of these approaches. Multiple years of data from northern rural Bangladesh show that, in any given year, about a third of poor households in the area send a family member to labor markets elsewhere in the country to cope with the seasonal shock through temporary, circular migration. This reliance on migration is not limited to Bangladesh and is common across the region. It is estimated that twothirds of the 740 million internal migrants in the world reside in Asia (World Bank 2013), and that India alone is home to an estimated 30 million temporary internal migrants (Deshingkar 2006). Our research focuses on seasonal migration, a type of temporary migration that occurs at a predictable time in the year (often-but not always-during the lean season) and lasts a few weeks to a few months. As a policy intervention, encouraging seasonal migration involves supporting the movement of people to where there are jobs and allowing the market to provide employment and income, which may be more efficient and easier than bringing jobs to local areas (as in a workfare program). Seasonal migration is a form of spatial arbitrage in which people move from areas where there are few jobs during a given season (e.g., due to lean periods in the agricultural crop cycle) and into areas with better employment opportunities during the same period. Seasonal migration not only diversifies income but may also have an indirect effect on other informal coping mechanisms. For example, the temporary migration of a subset of members in a risk-sharing network may have a spillover effect on others in the network through a system of gifts and transfers (). Lowering the cost of migration via grants or loans, as we do in our interventionbased research, may also make it easier for others to migrate because they can travel together with grant recipients as they share costs and risks. In the next section, we will delve into the details of the research methods and lessons from our multiyear, multisite research program on encouraging seasonal migration. III. Evidence on Seasonal Migration While our research program encourages more migration, one-third of poor households in rural areas of northern Bangladesh (in particular, the Rangpur region) already rely on internal seasonal migration as a coping strategy to deal with deprivation during the lean season. Those households send a migrant for weeks or months at a time for employment in urban areas or other rural areas within the country where wages are higher or employment opportunities are more broadly available or both. Data from 2017, for instance, reveal that the median daily wage among individuals in our study was Tk200 at home ($2.4) during the lean period, but Tk333 ($4) for those who migrated domestically. The starting point for our research was that the prevailing seasonal migration rate of one-third actually seemed puzzlingly low: given the lack of jobs in rural areas during the lean season, the relatively ample availability of jobs in urban areas during the same period, higher average wages offered to low-skilled workers in common destination areas, and the evident feasibility of temporary travel in this context, economic theory would predict a higher seasonal migration rate in the absence of market failures (and as long as individuals were not already sorted according to their comparative advantage). Our research was therefore designed to examine market failures that might prevent poor households in northern Bangladesh from temporarily moving to where there are jobs and to explore whether an external intervention could help overcome those hurdles. Rangpur is one of the poorest regions in Bangladesh, and its rural residents are more reliant on agricultural income than residents of other regions. Rural Rangpur households, on average, derive 50% of their income from farming, compared to 28% for rural households elsewhere in Bangladesh (Khandker and Mahmud 2012). This heavy reliance on agriculture leaves poor households more vulnerable to seasonal fluctuations from crop cycles (predominantly rice in this setting), making seasonal emigration a sensible strategy during periods when agricultural jobs are scarce. International remittance receipts are also relatively low in Rangpur compared to other regions in Bangladesh, as it often takes a fair amount of money to migrate internationally. And while 32% of households in Rangpur participate in some type of social safety net (mainly food transfers), the benefit amounts are too low to adequately address seasonal deprivation (Khandker and Mahmud 2012). With high poverty rates, savings constraints, and low safety net transfer amounts, fluctuations in employment opportunities and income translate into substantial drops in consumption during the preharvest period. Using data from our research, Figure 2 shows that up to 20% of poor households in this setting regularly miss meals during the lean season, and fewer than 40% never face food insecurity. Unaddressed, this can have disastrous consequences for those already living close to subsistence, especially families with pregnant women and young children. Reduced consumption for up to 3 months of the year can adversely affect the physical and cognitive development of children, with attendant effects on learning, productivity, and future earnings. Since 2008, we have tested whether transport subsidies can facilitate out-migration and help poor rural households avoid drastic drops in consumption during the lean season, specifically by taking advantage of employment opportunities elsewhere in the country. And since 2013, we have partnered with Evidence Action, a nongovernmental organization (NGO) supporting and implementing evidence-based programs, to explore the viability and impact of a scaled-up version of this intervention. Working with Evidence Action's "No Lean Season" project, we went beyond the first step of measuring the direct effects of seasonal migration subsidies on targeted beneficiaries to exploring the potential effects of a large intervention under the same framework, such as the effects on destination workers, permanent migration, and beliefs and social norms. A. Direct Effects of Seasonal Migration Support on Beneficiaries In 2008, we implemented an RCT covering 1,900 households across 100 villages in Rangpur (discussed in further detail in BCM ). First, each village was randomly assigned into one of four arms: (i) villages offered a grant for seasonal migration, (ii) villages offered a zero-interest loan for seasonal migration, (iii) villages given information about migration and job opportunities and wages at the destination, or (iv) a control group. Next, within each village, 19 poor households were randomly selected for the study, so that each of the 19 households living in the same village was assigned into the same RCT arm. 6 In August 2008, those in the first treatment arm were given information on popular migration destinations and their prevailing low-skill wages and offered a grant of Tk800 ($8.5) conditional on the temporary migration of at least one household member in the upcoming lean season. This amount was intended to cover the cost of a bus ticket and a few days of food, and in our data, we find that migrants spent on average Tk450 on migration transport in that round and Tk529 when including food and incidentals related to the journey (BCM 2014). 7 There were no further restrictions to the grant, and beneficiaries could choose the specific member(s) who would travel, the destination, the length of stay, and the nature of their job search. For the majority of the sample, we also did not impose any restriction on destination or the identity of travel companions. The second treatment arm was similar to the first, except that the disbursement was offered in August 2008 as a zero-interest loan rather than a grant. Repayment was expected upon return, but forgiven in the case of failure to find a job, though this latter feature was not disclosed upfront. The third arm only received information on popular destinations and prevailing wages using the same script as the first two arms, while the control group did not receive anything. This RCT design helped us evaluate whether the treatments were successful in encouraging more migration, which in turn allowed us to learn about the constraints limiting seasonal migration. We found that the small subsidy significantly increased the migration rate from 36% in the control group to 59% for those receiving the grant and 57% for those receiving the loan. The difference between the latter two is not statistically significant, while the average migration rate remained at 36% in villages with the information-only treatment. From these results, it appears that the relatively low rate of seasonal migration in the region, 6 Eligibility was defined as having less than 50 decimals (half an acre) of land and having skipped meals in the previous lean season. Of the households in these 100 villages, 70% were considered eligible (BCM 2014). 7 Migrants may have taken more than one trip or households may have sent more than one migrant, but the transfer amount is limited to Tk800 per household regardless of the number of trips. Calculating the overall cost of living at destination is problematic, as over half of migrants in our study receive housing and/or meals from their employer as part of compensation, or, in the case of rickshaw drivers, may be given housing above the garage. The monetary value assigned to these benefits differ by migrant, the quality of housing and food, and also from employers' perspectives on the value of the benefits they provide. despite the availability of jobs in urban areas, was not due to a lack of information about jobs-poor households either already know about the availability of jobs and prevailing wages in urban areas, or this piece of information is not relevant to the decision to migrate. To study the effect of the program in subsequent lean seasons, we tracked migration rates 1 and 3 years later, without repeating the intervention itself in those villages. During the lean season a year later, 47% of households in villages where conditional subsidies (loans or grants) had been offered in 2008 sent a seasonal migrant, which was significantly higher than the migration rate in the control and information groups. The migration rate in the grant and loan villages was still significantly higher (by 7 percentage points) than the control group even in 2011, the next year we collected data on this sample. Using the randomized treatment as an instrument, we find that households that were induced to migrate through this program increased both food and nonfood expenditures by 30%-35% relative to the control group, and consumed 550-700 more calories per person per day. This is equivalent to each person in the house eating one additional meal per day, during a period when food is scarce and meals are commonly skipped. Compared to the Tk800 transfer, households that sent a migrant in response to the subsidy increased their consumption by Tk350-Tk400 per person per month (BCM 2014). In a context where migration lasts 2-3 months and households have between four and five members, this represents a large rate of return on the initial subsidy. Figure 3 illustrates how the subsidies work, enabling households to send a migrant for temporary employment elsewhere in the country and using the additional earnings to raise consumption and calorie intake. Further analysis of our data indicates that poor families close to subsistence were hesitant to invest in the uncertain returns to migration partly because they were too risk averse. Even though seasonal migration is profitable on average-as indicated by the average gains in expenditures and consumption for households that responded to the subsidy by migrating-any chance of migration failure (e.g., spending the money for transport but failing to find a job, or at least one that is worth the travel) can have devastating consequences for households living at the edge of subsistence. As a result, even with knowledge about average benefits, poor households may be unwilling to risk spending the little money they have on migration. The migration subsidy acts as insurance against this downside risk, protecting households from having to use their own money upfront to pay for migration. B. Further Exploration for Scale-Up Results from the initial RCT show that providing financial support for seasonal migration has potential as a tool for addressing seasonal poverty. It is certainly cheaper to implement than work guarantee programs (which require. Seasonal Migration Subsidies in Practice Note: The subsidy amount has varied across rounds, from 800 taka to 1,500 taka ($8.5-$19, with variation in the exchange rate over the years) to respond to rising costs in migration as well as improved knowledge on the actual costs associated with migrating, which may include prepurchasing food for the family before migration. Source: Figure was directly provided by Evidence Action. paying beneficiaries for each day of work), owing to the fact that a one-time transport subsidy allows beneficiaries to earn market wages on their own at the destination. It is also more easily scalable considering the complexities of program implementation: the number of potential direct beneficiaries depends on the capacity of an institution to make subsidy disbursements but does not require the creation and management of jobs and payments. The upper bound of scalability may be constrained by the capacity of potential destination areas to absorb temporary laborers. But before declaring that support for seasonal migration should be scaled up and implemented as a policy for addressing seasonal poverty in Rangpur or other agrarian areas of Asia, the potential indirect effects and spillover effects of this intervention demand some attention. To move from an RCT to a policy, it is not sufficient for an RCT to simply demonstrate positive effects on its treated sample, particularly for a complex program that can in theory have repercussions for other parts of the population. When considering implementing an intervention as a large-scale policy, we must also look at the effect of the intervention on, say, other poor households who are operating in the same labor markets and competing for the same jobs. It is also important to understand any unintended noneconomic consequences on social norms, political beliefs, or intrahousehold decision-making. In contemplating a move from pilot-scale research to an at-scale program, programmatic issues also arise or gain more importance, such as the incentives and constraints faced by funders and implementers and the cost-effectiveness of the intervention. In the following subsections, we discuss these additional areas we have studied, providing information on the potential of this particular intervention as a scalable policy while also illustrating some issues that come into play in the move from RCT to at-scale program. 1. Targeting and Welfare The research described above focuses on the economic returns to migration, but there is also the question of whether migration raises welfare more broadly. If the extra income and consumption from migration comes at the cost of temporary family separation, worse living conditions for the migrant in urban slums, or any other negative experience stemming from migration, the effect of the subsidies on the welfare of targeted households cannot be represented simply by their income or consumption gain. Much of this "disutility" is difficult to directly observe or collect through survey questions. LMW use a simulated method of moments approach to implement a model that allows for unobserved disutility and matches this model to experimental moments generated from the same RCT reported in BCM. In the initial RCT described above (implemented in 2008), roughly 80% of migrants experienced success at the destination in economic terms, but at most 50% chose to remigrate in subsequent years. This gap between "success" (in terms of employment, income, and consumption) and the revealed preference of remigration choices is informative about how large the unmeasured disutility must have been, as a sizable portion of migrants who experienced success according to our economic metrics nonetheless choose not to migrate again in a subsequent year. The quantitative model in LMW matches these moments in the data to infer the disutility and the net welfare gains from migration. Through this method, LMW deduce that migration comes with substantial disutility, and that the actual welfare gains from the subsidies are smaller than the 30%-35% consumption increase observed through the experiment. We validate this inference with discrete choice experiments on the same BCM experimental sample, asking potential migrants to choose between migration and stay-at-home options that vary in the (hypothetical) conditions associated with the migration experience such as wages, the likelihood of finding work, living conditions at the destination, and the length of family separation. We learn that, of these dimensions, the quality of living conditions in the city is the component of welfare that matters most to migrants. In this context, toilet type and access serve as realistic proxies for living conditions, and we find that having housing with an indoor latrine (as opposed to public options) at the destination leads to a 17 percentage point increase in the reported propensity to migrate. This large effect is equivalent to increasing destination wages by 21%, or increasing the likelihood of finding work threefold. The clear policy implication is that governments should invest in urban living conditions and improve housing and sanitation in slum areas, as this would be valuable not only with respect to human rights and dignity but also for its economic draw. In a survey of migrants working in construction jobs in Dhaka, 71% report worse housing conditions at the destination compared to their own homes in rural areas (). Given the disutility from worse living conditions, this model suggests that potential migrants would only choose to leave when they are sufficiently economically desperate and the situation at home is precarious. The LMW modeling exercise allows us to learn about the types of households that are most likely to respond to the migration support intervention and the conditions under which they would choose to migrate in any given year. Understanding who exactly this migration support program manages to target is an important component of understanding the economic growth or welfare generated by this program. It could have been the case that there are many rural workers who would fare well in the city but who are "spatially misallocated" in rural Bangladesh because they are not sure about their prospects in the city and do not travel as a result. The financial incentive for migration would allow them to try out the urban labor market, and those who then learn that they have a comparative advantage in the city become repeat cyclical migrants or even permanent migrants. The results derived through the LMW model suggest that this is not the case, likely because those with a strong comparative advantage in the city are for the most part already there. Instead, it is those who have experienced recent bad shocks but are hesitant to draw down their small savings to make another trip (that may fail) that are induced to migrate by the financial incentive. The migrants from these households are not necessarily a lot more productive in the city compared to the village, but the migration support program allows them to weather unexpectedly bad periods by accessing jobs in urban areas. As such, this program does not generate much economic growth, as workers are not generally spatially misallocated and moving them does not create substantial changes to aggregate productivity. Rather, the value of the program lies elsewhere: it offers a safety net to extremely poor households, enabling them to cope with cyclical and idiosyncratic shocks. This is also an efficient mechanism for targeting support to such households. The migration requirement acts as an ordeal that, in effect, aids in the selection of households who really need to travel that year, due to dire circumstances or an adverse shock they experienced at home. It is precisely through the disutility of migration that the program is able to better target those who can benefit from migration but are unable to afford its risk, compared to other possible programs we have discussed in this paper: (i) UCTs, which target participants much more broadly-often based on fixed assets-are vulnerable to leaks to the nonpoor and cannot easily identify those who have faced recent shocks; (ii) agricultural insurance, which typically targets richer, landed households; and (iii) weather index insurance for landless households (Mobarak and Rosenzweig 2013), which can only indemnify aggregate shocks and not protect households from uniquely harsh idiosyncratic shocks. Labor Market Spillover Effects in Sending Communities As a migration support program is scaled up, with offers made to more and more poor households, it may start indirectly affecting other poor households who live in the same communities as program participants, but who may themselves not receive the transfer or may not be able to send a migrant. In other papers, we investigate such "general equilibrium effects" of the migration support program through labor market channels and risk-sharing channels in villages where the program is implemented. ACM report on an expanded trial implemented in 2014 that randomized 133 study villages into three arms: (i) a high-intensity treatment in which around 70% of eligible households were offered the migration subsidy simultaneously (47 villages), (ii) a low-intensity treatment in which around 14% of eligible households were offered the migration subsidy (48 villages), or (iii) a control group (38 villages). 8 This setup creates five types of study households: offered a subsidy in villages where many others were also offered the subsidy; offered a subsidy but residing in a village where few others were offered the same subsidy, not offered the subsidy but in a village where many others were offered the subsidy, not offered a subsidy and in a village where few others were offered a subsidy, and not offered a subsidy and in a village where no one was offered the subsidy either. The ACM analysis reveals strong network effects in migration decisions: while households in low-intensity villages who were offered the subsidy are 25 percentage points more likely to migrate than the control group, those in high-intensity villages who were offered the subsidy are 40 percentage points more likely to migrate. The difference between these groups is statistically significant, and both represent a very large jump in migration rates from a mean of 34% for the control group (type 5 households). Notably, even those who were not offered the subsidy directly but who reside in high-intensity villages (type 3 households) are 10 percentage points more likely to send a migrant relative to households in control villages. Migration decisions therefore appear to be strategic complements, as one is more likely to migrate if others within one's network are also migrating. Further analysis reveals that connections matter, as migrants frequently travel in groups with others from the same village. An operational implication of this finding is that a program offering migration subsidies in this context will be more cost-effective by targeting many people concentrated in fewer villages rather than few people spread out across many villages, taking advantage of the positive spillovers within villages and networks in migration decisions. 9 ACM also document a positive spillover on households residing in the same village who do not send a migrant. For every 10 percentage point increase in the emigration rate, agricultural wages in the village rise by an average of 2.2%. This benefits agricultural workers who do not migrate as well as migrants during the weeks of the lean season when they are home. The real effects of the program are, however, slightly smaller, as food prices also increase by 0.9% for that same 10 percentage point increase in the emigration rate. This rise in the food price index is largely driven by a rise in the local price of fish, as families with (successful) migrants increase their protein consumption, particularly by consuming more fish (BCM 2014). Risk-Sharing Spillover Effects in Sending Communities While ACM focuses on spillover effects through labor market channels, in a separate study (), we look at how a migration support program may affect local risk-sharing networks, particularly for households that are members of the same informal risk-sharing network as program beneficiaries and reside in the same villages. In theory, migration could erode these informal risk-sharing networks, as migrants who are exposed to a new labor market opportunity may choose to self-insure instead and exit the network. More generally, even if the drastic outcome of network exit does not happen, other members may need to offer migrants a larger share of the risk-sharing pie to keep them interested in participating in the network. In this sense, migration subsidies could have a negative spillover effect on nonbeneficiaries. Conversely, the new migration opportunity may improve risk sharing in the aggregate across the network, by providing some members of the network a new income stream that is less correlated with the village income stream. The network's sources of income as a whole become more diversified, and aggregate risk sharing improves. Using a structural model on our panel data of migrants across four rounds, the paper by Meghir et al. finds that the latter effect dominates: migration opportunities not only weaken the link between own income and own consumption for those who migrate, but they do so for others in the village as well. That is, program villages exhibit higher levels of risk sharing after the treatment compared to control villages. Household consumption levels for both migrant and nonmigrant households in program villages become less volatile and less dependent on the household's own income. Households evidently diversify their income through migration and choose to remain in the network, consequently sharing the benefits of this diversification with other households in the village. This is a positive spillover: both direct beneficiaries and other village residents are better able to smooth their consumption through migration opportunities. Noneconomic Effects of Migration: Changes to Social Norms It has been documented that the permanent movement of people over long distances changes social or behavioral norms-or both-among both migrants and their host communities through pressures such as assimilation, adoption, and backlash (World Bank 2011). In northern Bangladesh, seasonal migration is generally characterized by the migration of just one member while the rest of the family stays home. Most of these temporary migrants are male heads of households (over 80% of all migrants in our sample) who are away for 2-3 months at a time, and the majority of households are nuclear, with only two adult members. During their absence, migrants may be exposed to different lifestyles, norms, ideas, and ideals, which could in turn transform their beliefs and actions once they return home. At the same time, during this period, women in nuclear households may take on more decision-making roles with respect to the family and the home, a shift that can also in theory have a persistent effect on the way the household is managed even once the migrant is back. We explore these possible changes in a study by Mobarak, Reimo, and Thachil and find that migrants do become more progressive in their beliefs: individuals offered migration subsidies in treated villages become 2 percentage points more likely to recognize that women are capable of managing a household on their own, an effect best attributed to wives (or other female household members) effectively taking on that role during a male migrant's absence (as opposed to migrants simply observing other women outside the family doing so, either in destination areas or in one's village with heightened male out-migration). When migrants are away, there is also a substantial shift in decision-making roles. While men clearly dominate decision-making when they are at home, the proportion of women reporting that they participate (alone or jointly) in decisions regarding household expenses triples for periods in which the migrant has traveled away. These changes in beliefs and experiences are statistically significant even after adjusting for multiple hypothesis testing. Individuals in treated households also tend to take on some other more progressive views with respect to society, such as agreeing with the notion that governments should address income inequality and rejecting vote buying by political parties. These changes, however, do not appear to translate into a difference in actions when migrants return home. Even though women take on additional responsibilities during the male migration period and migrants are more likely to recognize women's capabilities upon their return, there are no significant differences between treatment and control villages with respect to women's participation in household decisions-including those regarding their own physical mobility outside the house-once migrants return. We also see no difference in the use of social services or in civic participation, or in behaviors tied to gender norms, such as female labor force participation or expenditure allocations. It appears that, in this setting, the (perceived) social costs for deviating from the social norm are so high that migrants do not change their behavior even after adopting more progressive beliefs (Mobarak, Reimo, and Thachil 2018). While it is disappointing that we do not see any positive effect of seasonal migration toward more progressive and inclusive behaviors, it is also reassuring that we do not detect any negative effects, such as the deterioration of norms or a backlash. We observe these (non) effects in the short run (a few months to 2 years after offering migration subsidies), and it remains to be seen whether the observed changes in beliefs translate into broader changes in social norms regarding women's role in society as momentum builds and individuals learn about each other's (transformed) beliefs (Bursztyn, Gonzalez, and Yanagizawa-Drott 2018; Dhar, Jain, and Jayachandran 2019). Effects on Intimate Partner Violence In 2015, the Bangladesh Bureau of Statistics carried out a survey on violence against women, uncovering that 75% of women in rural Bangladesh have experienced some form of intimate partner violence (IPV) in their lifetime, and 28% experienced physical or sexual violence in the 6 months prior to the survey (Bangladesh Bureau of Statistics 2016). Mobarak and Ramos explore how subsidies to seasonal migration affect the likelihood of IPV in practice, particularly in light of the various forces through which seasonal migration can influence IPV in theory. There are three competing forces shaping the potential effect of migration on IPV. First, limited resources can raise conflict within the family, especially in contexts with traditional gender roles, where men are expected to provide most of the financial support for their family and failure to do so can be seen as failure in a broader social role. Poor households facing seasonal fluctuations in income may be particularly vulnerable to this type of conflict. A positive income shock through migration (and its subsidy) may reduce these poverty stressors and, with it, decrease the incidence of violence. Alternatively, increases in male income can strengthen their bargaining position, increasing female relative vulnerability and, potentially, susceptibility to violence. Lastly, reducing the time a woman spends with her potential perpetrator can itself decrease the risk of victimization. Since migrants are overwhelmingly male in Bangladesh, subsidies that encourage migration can effectively decrease women's exposure to their spouses-and potential abusers-mechanically leading to less overall IPV experienced by women in treated households. Using data from the 2017 RCT and focusing on the effect of the migration of married male heads of households, Mobarak and Ramos find evidence that subsidies to migration may decrease IPV, particularly by reducing women's exposure to their male heads of households. Relative to the control group, women in households who are offered the migration subsidies are 3.5 percentage points less likely to say they have experienced physical or sexual violence in the 6 months prior to the survey (a 10% decrease in incidence). The results are most consistent with the idea that seasonal migration has an additional mechanical benefit, giving women some temporary relief by physically separating them from their perpetrators. Whether this produces any persistent reductions in violence beyond the period of migration, including perhaps through a shift in gender norms over time, is still an open question. Long-Term Effects on Permanent Migration In Chowdhury, Mobarak, and Reimo, we explore whether seasonal migration leads to permanent migration-as migrants build networks in and gain familiarity with the city-or instead makes rural living more viable and permanent moves into the city less likely. For this analysis, we use a follow-up survey of all households included in the 2008 study, gathering information on the whereabouts of each member 8 years after the initial treatment. Household members or, in their absence, their neighbors were interviewed, producing a dataset with very low attrition-we do not know the migration status of less than 1% of households. Overall, we find relatively low levels of permanent out-migration from rural Rangpur in general. Over an 8-year period, only 5% of households in the BCM sample (aggregating across treatment and control villages) permanently migrated away from their home village. This is consistent with other district-level data that document comparatively low levels of out-migration from the northern part of Bangladesh, particularly relative to the southeast (UNFPA 2016). 10 Moreover, it is no more difficult for us to find the original 2008 sample households from the treatment villages than it is to find those from the control villages, or to learn of their whereabouts. There is also no significant difference in the likelihood in permanent migration of the household or one of its members between treated and control villages. The data are precise enough to rule out large effects: the seasonal migration subsidies induced at most one in 200 households to permanently migrate over an 8-year period after the offer. For the subsample of households that were primarily engaged in agriculture at baseline, the subsidies actually decreased their likelihood of permanent migration relative to the control group, with households in treated villages 3.8 percentage points less likely to permanently leave the area than those in control villages. 11 This indicates that, rather than serving as a gateway to permanent migration, seasonal migration actually makes living in rural areas more viable in the long run, particularly for households whose skills and experience are in agriculture. For households with a comparative advantage in agriculture, a one-time support for seasonal migration helps them draw on it as a coping strategy in subsequent years and thereby avoid the more costly and drastic coping strategy of permanent migration. For policy makers in Bangladesh concerned about overcrowding and congestion in Dhaka and other populated urban areas, this finding implies that supports for seasonal migration may be a tool for easing urban pressures that come from permanent rural-to-urban population influxes. Other Open Questions In contemplating the implementation of a migration support program at this scale or larger, one must also be aware of the spillover effects on poor households in destination areas-and potentially other rural villages. We designed the most recent implementation rounds (2017 and 2018) to capture these economic and noneconomic spillover effects, as loan offers are made to well over 100,000 households in each season (compared to fewer than 1,500 in the 2008 study). We initially expected the 2017 results to provide some information on general equilibrium effects, revealing the effect of seasonal migration subsidies on the employment prospects and earnings for would-be construction workers and rickshaw drivers (two of the most popular jobs for seasonal migrants in our research rounds when they reach urban areas) already living in destination areas. Unfortunately, however, the 2017 intervention did not lead to a statistically significant effect on migration, contrary to all previous rounds. This was likely due to a confluence of factors-both avoidable and unavoidable-such as having disbursement targets for each migration officer set too low, heavy workloads, and the worst flooding in the region in over 40 years (for more information, see Levy and Sri Raman 2018). 12 To better understand these results and to learn about the effect of seasonal migration subsidies on poor households outside target villages, the 2018 round was implemented with a very similar design as the previous year-though of course addressing some of the weaknesses encountered in 2017. Although we do not yet know the effect of the 2018 intervention on the seasonal migration of those in treated villages, we do expect the impact of these subsidies, even at over 85,000 direct beneficiary households, to be relatively small on residents of popular destination areas, particularly Dhaka. In the past rounds, migration destinations were quite varied, with less than 25% of all migrants traveling to Dhaka, even though it is the single most popular destination for migrants from Rangpur (and elsewhere in the country). Dhaka is home to 14 million people, so even if one-quarter of all loan recipient households send a migrant to the megacity, this would amount to less than 0.2% of its regular population. Nevertheless, it is possible that while this influx is negligible for the general population in Dhaka, individuals engaged in particular sectors popular with migrants-namely, construction and rickshaw pulling-do feel an effect, positive or negative (depending on whether seasonal migrants' labor are complements or substitutes to local labor). We expect the results from the 2018 intervention to provide more information on this potential spillover effect on destination workers, which becomes relevant as we move from pilot to at-scale program. Another consideration relates to the effect of seasonal migration on agricultural employers. We have found that, in the short run, inducing temporary migration out of rural areas increases local agricultural wages (ACM 2017). While this is beneficial to poor rural workers who do not migrate (as well as those who do migrate on the weeks they are home), it also imposes a cost on agricultural employers in the same villages, who must now offer higher wages to secure the labor they need. In the short run, this results in a pecuniary transfer from comparatively richer employers to poorer employees, reducing inequality in treated areas. However, landed employers are also more likely to be a politically powerful group, so that their losses can potentially pose a risk to the sustained implementation of this intervention. They may also choose to shift toward more labor-saving technology, with both theoretically positive and negative consequences for local residents. In Nepal, we are planning to test a way to guard against negative political risk, by designing an RCT that offers agricultural employers subsidized access to a labor-saving technology in the same villages where migration subsidies are provided. By conducting an intervention that deals with demand and supply simultaneously, we expect to learn about labor market interactions as general equilibrium effects come into play in response to the subsidies. Comparisons to Other Programs A program offering seasonal migration subsidies is one among several potential antipoverty interventions, and a part of our research agenda has been designed to study its merits and effects relative to other interventions in this group. Through an RCT in Indonesia (), we directly compare the effects of migration subsidies to a UCT of an equivalent amount. The results from this RCT implemented in West Timor (Nusa Tenggara Timur province) in 2017 reveal that only 13% of households who are offered the UCT migrate in the 6 months following disbursements, compared to 42%-46% when the transfer is made conditional on migration. The lower effect on migration from the UCT is not in itself surprising, but highlights the fact that lack of funds is not the only (or even main) factor limiting households' migration decisions. LMW take a completely different route for comparing effects between subsidies for seasonal migration and UCTs, combining data from the 2008-2011 interventions with a structural model to simulate effects. The simulations indicate that a UCT of the same amount as the migration subsidies would increase migration by less than 1% among poor households in Rangpur, and its welfare benefits for the poorest quintile of households would also be slightly lower. The simulations indicate that a one-time migration subsidy improves welfare (measured as consumption over a lifetime) of the poorest quintile by 1%, whereas a budget-neutral UCT program would improve it by 0.9%. This difference is driven by the fact that migration subsidies rely on self-targeting: only households who have faced negative shocks and are desperate for money take up the CCT and migrate, while the UCT transfers are applied uniformly to all households. In contrast to these two policies, a rural workfare scheme increases welfare of the poorest quintile by only two-thirds as much, or 0.6%, as it discourages households from migrating to locations that offer better wages. We have also directly compared the cost-effectiveness of seasonal migration subsidies to other existing transfer programs in Bangladesh using secondary information, as discussed in subsection II.B. As with seasonal migration subsidies, the majority of beneficiaries for each of the three existing pro-poor programs (food transfer, food and cash transfer, and guaranteed work) are in the bottom three income deciles. Ahmed et al. estimate that the food transfer (IGVGD) and the food and cash transfer (FSVGD) programs each increase consumption by five times as much as the workfare program per dollar spent. By our calculations, the seasonal migration support program is even more cost-effective, increasing consumption on a per-dollar-spent basis by almost twice as much as the food and cash transfer program, which is the most cost-effective of the three (Mobarak and Akram 2016). And disbursing the offers as zero-interest loans rather than grants (which can be recovered and reused for the program in subsequent years) makes seasonal migration subsidies three times as cost-effective as the food and cash transfer program. IV. Implications for Asia For the last 10 years, our research on seasonal migration has been primarilythough not exclusively-in Rangpur, the most rural division in Bangladesh (UNFPA 2016). The potential for this intervention to improve the welfare of poor rural households vulnerable to seasonal fluctuations in agricultural income and employment opportunities, however, evidently extends well beyond the area, into other parts of Bangladesh and the Asia and Pacific region more broadly. In this section, we discuss requirements and adaptations for seasonal migration subsidies as a concept and as a program, as well as the importance of context-in terms of both time and place-for its viability. A. Applicability and Adaptability Subsidies for seasonal migration may be relevant to many subnational areas throughout the Asia and Pacific region. In general, for such an intervention to have potential as a tool for addressing seasonal poverty, target settings must have three minimum characteristics: (i) An agricultural lean season. Recurring and predictable periods of hunger indicate that there are constraints that keep consumption and income tied too closely together and that existing coping mechanisms and support systems are not adequate for weathering drops in income. (ii) Households that find spending on migration risky. Supporting seasonal migration makes sense if there is a large rural population living close to subsistence and for whom spending money on migration but failing to earn enough to offset costs could be catastrophic. Seasonal migration subsidies lower the cost of failure to find a job in destination areas and enable would-be migrants to set aside money or goods as a cushion for their families prior to migration. (iii) Jobs available in nearby areas. Ideal conditions include the availability of employment opportunities in several urban areas 4-8 hours away. For any distance closer than this, rural residents can commute and probably do so, and larger distances may require much higher subsidies. The existence of multiple destination areas also makes it more likely that an increased influx of migrants can be absorbed without large impacts on the target labor market(s). Many rural areas around the world match these conditions, though not all. We conducted exploratory work for a potential replication in Malawi and Zambia, for example, but decided not to test the program further in either setting because we were not convinced that they met the third condition. It is not clear that either country has vibrant urban labor markets with labor demand that can absorb many domestic migrants. Instead, we chose to pilot and test a version of this intervention in West Timor, Indonesia, a setting where most poor households in rural areas are not landless (as is the case in Rangpur). This changed one fundamental aspect of the program design. During the preharvest period, poor rural household members in West Timor generally have to stay home and work on weeding and land management of their own farm and are not interested in migrating at that time-even though, like in Bangladesh, that is the period of seasonal deprivation. In response to this, we adapted the intervention to allow for migration in other periods. Although poor households have relatively more cash after a harvest, this period tends to also be best for migration among landowning poor households since there is little agricultural work to be done. In this study site, landowning migrants traveled after the harvest, aiming to save money in anticipation of the next lean season. When considering the implementation of migration subsidies in a new context, there may be other norms and processes that need to be accounted for as well, such as the prevailing labor migration arrangement. For example, a survey of construction workers found that in Bangladesh, only 10% of workers that secured a job through a contractor received any payment in advance, whereas 52%-80% of workers in India received advance payments. These advances facilitate migration, helping migrant households purchase goods to be left at home and guarantee some consumption during migration (), and may change the role of transfers or the amount required for migration subsidies. A migration support program in India may also affect welfare along a different margin: wages for workers who secure jobs at destination areas tend to be higher than for those who are recruited and receive advances from contractors, so that a migration subsidy may serve as an alternative to the use of contractors, enabling individuals to arrange their travel first and search for a higher-paying job upon arrival at their destination. Nevertheless, it is clear from our experience in Indonesia that subsidies for seasonal migration are viable solutions outside of Rangpur and that the program can be adapted to a certain extent to local conditions. Dimensions over which seasonal migration subsidies could be modified to account for circumstances include-but are certainly not limited to-transfer amounts, timing, modality (e.g., loans, grants, or even transport tickets), number of members to whom subsidies are made, and even the extent to which the program works with employers or contractors and facilitates hiring in advance. Some of these modifications are more drastic than others, and a few might require piloting and further testing, but none alter the fundamental nature of the concept, which is to support and subsidize the temporary movement of people from rural areas to destinations where there are more job prospects for low-skilled workers. B. Limitations Drawing a distinction between subsidies for seasonal migration (the concept) and "No Lean Season" (the program), we recognize that while the former has a broader potential and applicability than the latter, neither is implementable everywhere. Not only are seasonal migration subsidies likely not a worthwhile investment in settings where one of the three requirements above are not met, but the extent to which subsidies can induce migration is highly context dependent. In 2013, we attempted to implement the first version of "No Lean Season" after the positive results of the initial RCT on seasonal migration. That same year, however, mass political strikes (hartals) "designed to disrupt the county's transportation network" and that involved the burning of buses (the main mode of transportation used by migrants from Rangpur) took place throughout Bangladesh (Ahsan and Iqbal 2016). As one of the main goals of this form of protest was to "restrict vehicular movement in key urban areas," it naturally led to longer transport times and higher costs during that period, not to mention fear among the population in and outside the cities (Ahsan and Iqbal 2016). While the lack of takeup in response to our migration subsidy offers under these conditions might not be surprising in hindsight, it also points to an implementation challenge for such a program. As strikes are often unpredictable more than a few weeks in advance, implementation of the program-from contracting to loan offers-may already be underway when it becomes clear that circumstances will limit take-up. We witnessed a similarly low take-up in 2017 and suspect that extreme flooding in northern Bangladesh-the likes of which had not been seen in 40 years-contributed to this pattern. In Rangpur, the yearly swelling of rivers can lead to houses or entire villages being swept away, but that year's uniquely severe flooding made implementation of the program especially difficult and may have also discouraged seasonal migration, as potential migrants cannot abandon their families under likely disastrous weather conditions, and transport may be more limited or precarious. External shocks such as extreme weather conditions and violence impose limitations on the applicability of seasonal migration subsidies as a concept, indicating that even in settings where the intervention would normally work, it might not work in certain years, no matter the design. This is distinct from-and in addition to-programmatic limitations related to implementation capacity and delivery. In the transition from a closely delivered and monitored RCT during 2008-2011 to "No Lean Season," the implementation of the program was handed over to a local partner, RDRS Bangladesh, a microfinance institution. We have learned that its decades of experience in microfinance and in the region are both a benefit and a hurdle, as its managers and officers are used to thinking in terms of loan disbursement and repayment and its institutional measurements are tailored around that model. But if one were to implement "No Lean Season" as a pure microcredit program, it would make sense to focus efforts on individuals most interested in migrating and most likely to repay-not necessarily those who recently experienced a drastic negative shock. While we believe that flooding played a role in the 2017 migration rates, we suspect that part of this dampened result also stemmed from targeting issues, as migration officers were given disbursement goals by their managers and may have concentrated their efforts on households most likely to migrate anyway. In response to these findings, RDRS Bangladesh changed the goals set for its officers in 2018. Most recent loan disbursements show a promising change, with twice as many loans made per branch compared to the previous year ( Figure 4). It is also worth noting that, while the intervention in Bangladesh has been transformed into "No Lean Season," migration subsidies can in theory be implemented not only outside this program but also without an NGO or microfinance institution entirely. The concept could be taken on by national or subnational governments for whom cost-benefit calculations may weigh differently. A program that might not be feasible from the perspective of an NGO, which typically requires a minimum impact per dollar, may nevertheless be worthwhile to a government for its benefit to the rural poor in addressing seasonal deprivation. V. Conclusion The last 10 years of research on seasonal migration has shown that an intervention that supports this strategy by subsidizing travel can have large positive impacts on poor rural households. In several study rounds conducted in northern Bangladesh, households that were offered a small migration subsidy were 22-40 percentage points more likely to migrate over a given lean season compared to control households. Households that responded by sending a migrant also recorded on average higher levels of consumption, income, and expenditures during the lean season than those not offered the subsidy. Villages in which a large share of poor households were offered the migration subsidies also experienced an increase in wages during this period, which is usually characterized by low employment opportunities and pay. A one-time migration transfer continued to have a positive effect on migration rates up to 3 years later. We believe that this type of intervention may be an appropriate tool for poverty alleviation-particularly seasonal poverty-in rural areas where a large part of the population (both in relative and absolute terms) is engaged in agriculture and lives close to subsistence for at least part of the year and where potential destinations with ample low-skill temporary employment opportunities are within a reasonable travel time (4-8 hours). The complete lack of seasonal migration is not a required element, as exemplified in Rangpur, and potential beneficiaries are not restricted to traveling to destinations 4-8 hours away. Among migrants in our study in northern Bangladesh, for example, migration is already fairly common (but still lower than expected given the availability of jobs elsewhere in the country), and Dhaka is a popular destination despite being a day's travel away. In India, temporary migrants are just as likely to migrate to urban centers within their state as to other states, and they are actually less likely to migrate within their district (Imbert and Papp 2019). There are other programmatic requirements for the success of a migration subsidy intervention, a topic that we have not covered in depth here. Local implementation capacity and political support are likely crucial. In Rangpur, our intervention has been implemented through RDRS Bangladesh, a local institution with a strong presence and long history in the northern part of the country since 2014. In Indonesia, we have experimented with collaborating with local government agencies. In general, the nature of the implementing partner (e.g., local NGO, international organization, or government) may be less important than its capacity, will, and presence in rural areas, as the design of the intervention offers some flexibility to adapt to an implementer's priorities and requirements. Our research results also highlight the importance of removing other types of barriers to temporary migration. The PRC, for example, imposes many explicit restrictions on the movement and employment of rural workers. Other countries in the region also have policies that implicitly deter poor rural households from taking advantage of urban labor markets. In India and Viet Nam, for example, full access to social safety nets is given only to individuals at their permanent residence area (Deshingkar 2006), which limits poor households' willingness to send a migrant away in search of jobs elsewhere lest they lose benefits at home. Encouraging seasonal migration is a complex intervention that may produce a range of indirect effects and unintended consequences beyond the direct economic effects on treated households. Over the last 10 years, our research agenda has expanded from considering the effect of these subsidies on the migration, consumption, and income of targeted households to exploring the secondary effects on both beneficiaries and nonbeneficiaries, with scaling up always in mind. Results from the initial RCT in Bangladesh with 1,900 households only indicated that this intervention was "promising." The path from a successful RCT to an implementable program called for a greater understanding of potential general equilibrium, noneconomic, and long-term effects. The majority of the population in Asia remains rural and agrarian. Support for seasonal migration can play a valuable role in helping poor rural families cope with drops in employment opportunities and income during lean periods in the agricultural cycle. The Asia and Pacific region has a uniquely large concentration of poor households in rural areas, but it is also peppered with large urban areas and manufacturing zones that have attracted both domestic and regional migrants. While the applicability of seasonal migration subsidies may vary across and within countries-depending on the dominance of the agricultural cycle, proximity of poor households to potential destination areas, and the ability of an area to absorb temporary migrants-the basic elements of demand for this intervention exist in many parts of the region today. Lowering the barriers to temporary migration-through changes in policies (either explicit or implicit), investment in transportation networks, or subsidies, or a combination of these-can expand poor rural households' access to labor markets elsewhere in their countries. And while encouraging seasonal migration might not be a path to growth in contexts where migration is already common and rural residents are unlikely to be spatially misallocated, facilitating the free movement of people within their own countries-enabling them to take advantage of labor opportunities elsewhere and to avoid resorting to hunger-is a desirable pursuit in its own right. Poverty reduction in Asia has been associated with diversification away from farm activities-of which employment opportunities in urban areas is one possibility-as opposed to increasing farm productivity (World Bank 2012). Policies that support seasonal migration can help steer countries down that path, encouraging poor rural households to diversify their income sources and increasing labor supply for sectors with higher-productivity potential than agriculture. In fact, seasonal migration may be a valuable but temporary tool. As countries develop, transportation networks improve, and nonagricultural employment opportunities for rural residents expand at home, the need for seasonal migration as a coping strategy might decrease. Until then, supporting seasonal migration-through direct policies or interventions such as "No Lean Season," or both-can help address seasonal poverty and hunger in various parts of the region.

Human Lung Mast Cell Products Regulate Airway Smooth Muscle CXCL10 Levels In asthma, the airway smooth muscle (ASM) produces CXCL10 which may attract CXCR3+ mast/T cells to it. Our aim was to investigate the effects of mast cell products on ASM cell CXCL10 production. ASM cells from people with and without asthma were stimulated with IL-1, TNF-, and/or IFN and treated with histamine (1100M) ± chlorpheniramine (H1R antagonist; 1M) or ranitidine (H2R antagonist; 50M) or tryptase (1nM) ± leupeptin (serine protease inhibitor; 50M), heat-inactivated tryptase, or vehicle for 4h or 24h. Human lung mast cells (MC) were isolated and activated with IgE/anti-IgE and supernatants were collected after 2h or 24h. The supernatants were added to ASM cells for 48h and ASM cell CXCL10 production detected using ELISA (protein) and real-time PCR (mRNA). Histamine reduced IL-1/TNF--induced CXCL10 protein, but not mRNA, levels independent of H1 and H2 receptor activation, whereas tryptase and MC 2h supernatants reduced all cytokine-induced CXCL10. Tryptase also reduced CXCL10 levels in a cell-free system. Leupeptin inhibited the effects of tryptase and MC 2h supernatants. MC 24h supernatants contained TNF- and amplified IFN-induced ASM cell CXCL10 production. This is the first evidence that MC can regulate ASM cell CXCL10 production and its degradation. Thus MC may regulate airway myositis in asthma. Introduction In asthma, airway smooth muscle (ASM) cells may play a significant role in supporting inflammation locally. They produce a wide range of chemokines and growth factors in vitro in response to proinflammatory cytokines, which are elevated in asthma following release by activated inflammatory cells and other airway structural cells. Degranulating mast cells (MC) are present in higher numbers in the ASM bundles of people with asthma than people without asthma. MC numbers are further increased in allergic compared with nonallergic asthma and thus may be a specific feature of eosinophilic asthma. There is conflicting evidence about the presence of MC in the ASM in severe asthma, which is often associated with neutrophilia and steroid resistance. Carroll et al. found that degranulating MC numbers were highest in the ASM layer in fatal asthma, whereas Balzar et al. found no evidence of them in the ASM in biopsies from a severe asthma cohort. MC activation by allergen mediated cross-linking of IgE results in the release of a variety of preformed and newly synthesised mediators. These include the major granule-derived mediators histamine and tryptase and newly synthesised cysteinyl leukotrienes, whose effects on ASM contractility are well established. MC also produce other proteases, arachidonic acid metabolites, and a wide range of cytokines and growth factors. The relative balance of these mediators in the vicinity of the ASM cell will determine the overall effect MC have on ASM production of a certain chemokine. What role that chemokine plays in airway inflammation will depend on its quantity and activity locally. To date, the effects of many MC products on ASM chemokine production are unknown. When studied individually, some MC mediators directly affect ASM chemokine production and thus may regulate inflammation locally. For example, interleukin (IL-)1 or tumour necrosis factor (TNF-) alone and IL-4 or IL-13 alone and in combination with IL-1, induce ASM release of the potent eosinophil 2 Journal of Allergy chemoattractant CCL11 (eotaxin), while we and others have shown that MC proteases cleave CCL11. We have also shown that histamine enhances IL-1 -induced granulocytemacrophage colony stimulating factor (GM-CSF) release but strongly inhibits CCL5 (RANTES) release by ASM cells from people with and without asthma. Further studies are needed to investigate the pro-and anti-inflammatory effects of MC and whether or not they can regulate their own recruitment to the ASM and survival there. Increased localization of MC to the ASM layer could be controlled by many ASM cell derived chemokines, as human lung MC (HLMC) express a range of chemokine receptors. However MC in the ASM of people with asthma are CXCR3 positive, whereas less than half of the MC elsewhere in the airway submucosa express CXCR3. Further, ASM in biopsies from people with asthma are often (∼50%) immunoreactive for the CXCR3 ligand CXCL10, whereas the ASM in biopsies from people without asthma are not. In vitro we have established that asthmatic ASM cells produce CXCL10 more rapidly than nonasthmatic ASM cells under Th1 inflammatory conditions and MC chemotaxis towards medium from the asthmatic ASM cells is driven by CXCL10 activating CXCR3 on the MC. Whether or not MC can regulate ASM CXCL10 production and activity is not known. Thus, the aims of this study were to investigate the effects of MC products on CXCL10 production by ASM cells from people with and without asthma. The effects of the granulederived products histamine and tryptase, as well as the overall effects of human lung MC products released in the first 2 h or 2-24 h after activation, on ASM CXCL10 production were examined. Airway Smooth Muscle. ASM cell cultures were established from lung samples donated by 16 people with a doctor diagnosis of mild to moderate asthma (mean age 36 y and range 23-62 y) and 17 people without asthma (mean age 55 y and range 29-83 y). The lung samples were either bronchial biopsies or resected lung tissue obtained from people undergoing surgery for thoracic malignancies or lung transplantation. All samples were obtained with the donor's informed consent and approval from Sydney South West Area Health Service or Australian Red Cross. Approval for this study was granted by The University of Sydney Human Ethics Committee. Airway Smooth Muscle Cell Culture. ASM bundles were dissected out from macroscopically normal lung samples and grown as explants. The cells from people who had no doctor diagnosis of asthma are referred to as nonasthmatic. ASM cells were maintained in culture as previously described in Dulbecco's Modified Eagle's Medium (DMEM) (Sigma-Aldrich) supplemented with 10% v/v heat-inactivated foetal bovine serum (FBS), 100 units/mL penicillin G, 100 g/mL streptomycin sulphate, 25 g/mL amphotericin B, 4 mM Lglutamine, and 20 mM HEPES, at pH 7.4 (growth medium) and grown at 37 ∘ C in a humidified 5% CO 2 in air atmosphere. The cells adopted a "hill and valley" growth pattern and were positive for -smooth muscle actin and h-calponin using immunofluorescence. ASM cells harvested between passages 4 and 7 were used in experiments. Histamine and Tryptase Effects on ASM Cell CXCL10 Production. To investigate the effects of histamine and tryptase on ASM cell CXCL10 production, ASM cells were seeded into the wells of 6-or 24-well plates at 1 10 4 cells/cm 2 in growth medium. After 7 days growth, the cells were serum-deprived for 48 h in FBS-free DMEM supplemented as described above and also with 0.1% v/v bovine serum albumin (BSA) (serum-free medium). The well cultures were left untreated or treated with histamine (1, 10, or 100 M), tryptase (0.2, 1.0, or 5.0 nM), or the vehicle for tryptase and heparin (50 M), as used previously. Immediately afterwards the cytokines IL-1, TNF-, and IFN, alone and in combination (cytomix), were added to untreated and treated wells. The cytokines were each used at 10 ng/mL and all treatments were performed in duplicate. To investigate which histamine receptor was involved in the effects of histamine on ASM cell CXCL10 release, the cells were treated in duplicate with either the H1 receptor antagonist chlorpheniramine at 1 M or the H2 receptor antagonist ranitidine at 50 M, for 30 minutes prior to the addition of histamine followed by the cytokines. To investigate whether the proteolytic activity of tryptase was involved in its effects on ASM cell CXCL10 release, an aliquot of the same tryptase batch was first heat-inactivated at 56 ∘ C for 30 minutes and then immediately added to the ASM cells, or the serine protease inhibitor leupeptin (50 M) was added to tryptase-treated ASM cells 3 h after the addition of the cytokines. After 4 hours cytokine stimulation, the cells in the well cultures were washed and total RNA was extracted using the guanidine thiocyanate/phenol chloroform method. The RNA was reverse transcribed using RevertAid First Strand cDNA Synthesis Kit (Fermentas Life Sciences, Hanover, MD, USA) and the cDNA amplified by PCR using FAM-labeled human CXCL10 and VIC labeled 18srRNA TaqMan probes on an ABI Prism 7500 (Applied Biosystems) as described previously. Alternatively, the ASM well cultures were incubated with the cytokines for 24 hours and then the culture medium (CM) was collected and stored at −20 ∘ C for later analysis of CXCL10 levels using the CXCL10 Duo-set ELISA kit and protocol (R&D Systems, Minneapolis, MN, USA). Tryptase Effects on Recombinant Human CXCL10. To investigate whether tryptase can cleave CXCL10, recombinant human CXCL10 diluted in serum-free medium to 250, 500, and 1000 pg/mL, or the same volume of CM from cytokinestimulated ASM cells, was added to a series of wells in microwell plates. Tryptase (1 nM), heat-inactivated tryptase, or tryptase followed by leupeptin (50 M) 3 hours later, was added to duplicate wells containing CXCL10 or the ASM cell CM and the plates were incubated at 37 ∘ C for 24 hours and then CXCL10 was detected using ELISA. Human Lung Mast Cell Isolation and Activation. MC were isolated from macroscopically normal lung tissue dissected immediately by a pathologist from lung samples donated by 6 people undergoing surgery for thoracic malignancies. The MC were isolated within 24 hours of resection as previously described. Briefly, the lung parenchyma was chopped finely, washed, and digested with collagenase (7.5 mg/g tissue) and hyaluronidase (3.75 mg/g tissue) for 90 minutes at 37 ∘ C. MC in the cell suspension were positively selected using sheep anti-mouse IgG-coated magnetic beads (Dynal, Oslo, Norway), which were coated with a second antihuman CD117 (clone YB5B8; Bioscientific, Sydney, Australia) antibody (5 g/mL) just before use and resuspended at 1 10 6 cells /mL in growth medium. The HLMC were activated with IgE (2.5 g/mL) and goat anti-human IgE (1 g/mL) (Calbiochem, CA, USA) immediately or left unstimulated. The HLMC culture supernatants (SN) were collected after 2 hours (MC 2 h SN) and fresh growth medium was added to the HLMC. This medium was collected 24 hours after the HLMC were first activated (MC 24 h SN). The MC 2 h and 24 h SN were stored at −20 ∘ C until immediately before use in the experiments described below. ASM Cell Treatment with Human Lung Mast Cell Products. To investigate the effects of HLMC products released at different times after activation on ASM cell CXCL10 production, harvested ASM cells were placed in the wells of 96-well plates at a density of 3200 cells/100 uL/well in growth medium and incubated as described above. After 24 hours the cells were serum-deprived in serum-free medium for 72 hours. Then the ASM cells were stimulated with growth medium ± IFN (10 ng/mL) and, after 30 minutes, treated in triplicate or quadruplicate with the MC 2 h or 24 h SN (prepared as described above) at 0, 20, or 40% v/v in growth medium in the presence of the same concentration of IFN, or left untreated. This order was used to ensure that the ASM cells were activated by IFN as it induces CXCL10 production and the effects of the MC SN directly on its activity were not known. After 48 hours of treatment, the medium was collected from each ASM culture and stored at −20 ∘ C for later measurement of CXCL10 levels using ELISA. ASM cell CXCL10 release in response to growth medium ± IgE/anti IgE was also determined. Protease Involvement in the Effects of HLMC Products Released by 2 h. To establish whether or not MC proteases were involved in modulating ASM CXCL10 release, 2 h MC SN or the same SN already pretreated for 30 minutes with 50 M leupeptin were added at 40% v/v in growth medium to the ASM cell well cultures. After 3 hours coincubation at 37 ∘ C, leupeptin was added to some of the wells containing untreated MC SN and the incubation was continued. All the culture medium was collected from each well after 48 hours and CXCL10 levels were quantified using ELISA. To investigate if the effects of 24 h MC SN on cytokineinduced ASM CXCL10 could be due to TNF-produced by MC, TNF-levels were measured in the MC SN using ELISA. Then serum-deprived ASM cells were stimulated with IFN-(10 ng/mL), with and without a similar range of TNFconcentrations to those detected in the MC SN, and made up in growth medium. After 24 h incubation at 37 ∘ C, the culture SN were collected and CXCL10 levels in them were quantified as above. The effects of 2 h MC SN, -tryptase, or leupeptin on the CXCL10 ELISA capture antibody were also investigated. Wells were coated with CXCL10 capture antibody and washed ready for use as usual; then either 40% v/v 2 h MC SN, 1 nM -tryptase, 50 M leupeptin, or vehicle, in growth medium or growth medium alone, was added to the wells already containing capture antibody. After incubation together for 1 hour at 37 ∘ C, the wells were washed and doubling dilutions of CXCL10 standards were added and the remainder of the ELISA protocol was completed without modification. 2.9. Data Analysis. CXCL10 mRNA or protein levels from replicate treatments were averaged. The results for each experiment were expressed as a percentage of the cytokine control. The mean ± SEM was then calculated for the asthmatic and non-asthmatic cell lines and for each treatment. Statistical analyses were performed on all data using Statview (SAS Institute, Cary, North Carolina) or Prism 5.03 (Graph-Pad, La Jolla, CA) and significance ( < 0.05) was determined using 1-way or 2-way analysis of variance (ANOVA) with the Bonferroni correction for multiple comparisons or using the Student's -test as appropriate. Histamine Effects on ASM Cell CXCL10 Production. In preliminary experiments we established that histamine did not induce ASM cell CXCL10 release. The levels of CXCL10 released by untreated ASM cells from people with and without asthma were very low (Table 1). Histamine treatment at 1, 10, or 100 M for 24 h did not alter CXCL10 release by the ASM cells (data not shown). Similar to previous reports, cytokine stimulation increased ASM cell CXCL10 production. IL-1, TNFand IFN, alone and in combination (cytomix), stimulated significant CXCL10 release from the asthmatic and nonasthmatic ASM cells (Table 1). Although IL-1 and TNF-both induced significant increases in CXCL10 gene expression over 4 hours, histamine 10 M did not affect CXCL10 mRNA levels induced by either cytokine (Figure 2(a)), even though it significantly reduced CXCL10 release. Further, the effects of 10 M histamine on CXCL10 release were not reversed by pretreating the asthmatic and non-asthmatic ASM cells with the H1 and H2 receptor antagonists chlorpheniramine and ranitidine, respectively, or with the cell membrane permeable adenylate cyclase inhibitor SQ 22,536 (Figures 2(b) and 2(c)). Tryptase reduced the levels of CXCL10 detected in CM from asthmatic and non-asthmatic ASM cells. When tryptase (1 nM) was added to the ASM cells 30 minutes after the cytokines IL-1, TNF-, IFN, or cytomix, the CXCL10 levels detected in non-asthmatic and asthmatic ASM cell CM were very much lower than the respective cytokine control ( Table 2). The heparin vehicle for tryptase did not affect cytokine-induced CXCL10 levels (data not shown). To investigate whether tryptase proteolytic activity was involved, the effects of tryptase and the same tryptase either heat-inactivated just before use or in the presence of the serine protease inhibitor leupeptin were compared. Untreated tryptase again markedly reduced nonasthmatic and asthmatic ASM cell cytokine-induced CXCL10 levels, whereas heat-inactivated tryptase did not affect them (Figures 3(a)-3(d)). Similarly, tryptase did not reduce CXCL10 levels when leupeptin (50 M) was added 3 hours after the cytokine(s). However asthmatic ASM cell CXCL10 levels were more variable in the presence of leupeptin. CXCL10 levels in CM from asthmatic IL-1 -, TNF--, or IFN -stimulated cells treated with tryptase followed by leupeptin were higher than levels in the presence of the same tryptase which had been heat-inactivated, but the differences were not significant (Figures 3(a)-3(d)). Tryptase Effects on Recombinant Human CXCL10. The proteolytic effects of tryptase on CXCL10 were investigated further in a cell-free system. Significantly less CXCL10 was detected in wells containing human recombinant CXCL10 (500-1000 pg/mL in serum-free medium) following treatment for 24 hours at 37 ∘ C with 1 nM tryptase than with its vehicle (Figure 4). CXCL10 levels were not reduced in wells treated with heat-inactivated tryptase and were even increased in those treated with tryptase followed by leupeptin (50 M) 3 hours later. Similarly, tryptase treatment aftercollection also markedly reduced detectable CXCL10 levels in culture medium collected from cytokine-stimulated ASM cells when compared to the vehicle-treated controls (data not shown). The capacity of the antibody from the Duoset kit to capture CXCL10 standards was not affected by direct incubation with tryptase, its vehicle, or leupeptin at the concentrations used above (data not shown). release in its presence over the 48 hour incubation. CXCL10 release was generally greater from asthmatic ASM cells (9.3 ± 2.4 ng/mL, = 6) than non-asthmatic ASM cells (5.4 ± 2.0 ng/mL, = 6), but the difference was not statistically significant. ASM cell CXCL10 release was not affected by the presence of IgE/anti-IgE ( Figure 5(a)). Effects of Human The MC 2 h SN markedly reduced asthmatic and nonasthmatic ASM cell CXCL10 levels following IFN stimulation. At 20% v/v, the MC 2 h SN significantly reduced nonasthmatic ASM cell CXCL10 levels down to 47 ± 12% of the FBS-IFN control and at 40% v/v significantly reduced asthmatic ASM cell CXCL10 levels to 46 ± 15% of control 6 Journal of Allergy ( Figure 5(b)). To investigate whether this effect was due to MC protease activity affecting early events leading to CXCL10 production or affecting CXCL10 after new gene transcription had commenced, leupeptin (50 M) was added at two different steps. Leupeptin inhibited the effects of the MC 2 h SN irrespective of whether the SN were treated with it before they were added to the ASM cells (0 h), or it was added to the ASM cultures 3 hours after the SN (3 h) ( Figure 5(c)). When the CXCL10 capture antibody was incubated with the MC 2 h SN there was no effect as detection of human recombinant CXCL10 in the ELISA remained the same as with the vehicletreated and untreated capture antibody controls (data not shown). In contrast, the MC 24 h SN increased asthmatic and non-asthmatic ASM cell IFN -induced CXCL10 release. The MC 24 h SN at 20% and 40% v/v significantly increased IFN -induced CXCL10 release from asthmatic cells up to 157 ± 28% and 137 ± 16% and from non-asthmatic cells up to 155 ± 22% and 170 ± 32% of the FBS-IFN control, respectively (Figure 6(a)). As TNF-acts synergistically with IFN to increase CXCL10 production, the levels of TNF-in MC SN were quantified. The MC 2 h and 24 h SN contained TNF-at 227 ± 79 and 434 ± 87 pg/mL, respectively ( Figure 6(b)). TNFover those concentrations increased ASM cell IFN -induced CXCL10 release by ≥5fold (Figure 6(c)). Discussion In asthma, the ASM produces the chemokine CXCL10 which we have previously shown induces MC chemotaxis through CXCR3 engagement. CXCL10 may also mediate CXCR3 + T lymphocyte recruitment to the ASM in severe asthma and following allergen challenge. Thus CXCL10 may be critical in asthma because it can induce airway myositis. This is the first study to examine the effects of MC products on CXCL10 production by ASM cells from people with and without asthma. The effects of individual granule-derived MC products and all the products released by human lung MC in the first 2 or 2-24 hours following IgE receptor activation were investigated. The key granulederived product histamine reduced ASM cell CXCL10 release induced by the proinflammatory cytokines IL-1 and TNF-. As well, the proteolytic activity of tryptase, or the leupeptinsensitive serine proteases released by human lung MC within two hours of activation, markedly reduced CXCL10 levels released following ASM cell cytokine stimulation. In contrast, human lung MC products synthesised and released later increased ASM CXCL10 release and levels of human lung MC-derived TNF-were sufficient to amplify IFN -induced CXCL10 production. These findings are evidence that mast cells are able to regulate ASM cell CXCL10 production and degradation and thus may regulate their own and/or T cell recruitment to the ASM in asthma. Following MC activation histamine, along with other products also stored in the granules, is released rapidly (reviewed in ). Histamine has a very broad spectrum of activities and histamine levels are high in the bronchoalveolar lavage fluid of people with asthma and correlate with airway hyperresponsiveness. The contractile effects of histamine on ASM are well characterised and mediated via its H1 and H2 receptors, which are highly expressed on ASM. Histamine binding to the H1 receptor primarily activates phospholipase C-inositol triphosphate-diacylglycerol signalling and leads to Ca 2+ release from intracellular stores and protein kinase C activation, both of which can also upregulate cAMP. Signalling via H2 receptors activates adenylate cyclase to directly generate cyclic adenosine monophosphate (cAMP), but H2 receptors are also coupled to the phosphoinositide system. Through the H1 receptor, histamine can also activate NF B and thus might be expected to enhance proinflammatory gene transcription. Histamine also modulates the activity of the coactivator cAMP response element-binding protein (CREB/CBP) to mediate some of its effects on gene expression. However in this study histamine reduced IL-1 -and TNF--induced CXCL10 release, but not gene expression, which is mediated via the transcription factor NF B. Neither the H1 nor the H2 receptor antagonists chlorpheniramine and ranitidine, respectively, prevented the inhibition of CXCL10 release. Nevertheless, differences in transcriptional pathways following ASM cell stimulation with the individual cytokines and cytomix may contribute to the differential effects of histamine indirectly. In contrast to ASM CXCL10 production induced by IL-1 and TNF-, IFN -induced production was not affected by histamine. Although Smith et al. demonstrated that IFN, but not TNF-, significantly increases histamine-induced inositol-phosphate signalling and H1R mRNA expression, those IFN effects do not match with the observations reported here. IFN activates the JAK 2 -STAT-1 pathway predominantly and NF B only weakly in nonasthmatic ASM cells to induce CXCL10 production. As observed here and previously, IFN synergistically increases CXCL10 production when used with IL-1 and/or TNF-, but Clarke et al. did not observe any effects on NF B or STAT-1 activation and DNA binding. Rather they found that the combined cytokines synergistically increase the recruitment of CREB binding protein (CBP) and RNA polymerase II to the CXCL10 promoter. Such IFN -induced increases in CREB binding to the CXCL10 promoter may overwhelm any inhibitory effects of histamine. Further, we have observed in asthmatic compared with non-asthmatic ASM cells that NF B activation is stronger and STAT-1 activation is weaker in response to the three cytokines combined (cytomix), which may underlie the small inhibitory effect of histamine in the non-asthmatic ASM cells. Interestingly CXCL10 production induced by IFN or cytomix is also not inhibited by current asthma therapies such as fluticasone and/or salmeterol, whereas IL-1 -and TNF-induced release is markedly reduced by these agents. Histamine and cytokines may have opposing effects on intracellular (Ca 2+ ) levels in human ASM cells. Mitochondria buffer cytosolic (Ca 2+ ) levels and histamine causes a rapid increase in both cytosolic and mitochondrial (Ca 2+ ) levels. Whereas TNF-decreases mitochondrial (Ca 2+ ) levels through the mitochondrial Na + /Ca 2+ exchanger, while increasing cytosolic (Ca 2+ ) levels. As indicated above, TNF-also induces NF B mediated CXCL10 gene transcription, whereas histamine does not. NF B mediated gene transcription is inhibited by SERCA pump inhibition, including CXCL10 production, especially in asthmatic ASM cells. If histamine modulated TNF--induced changes in mitochondrial and cytosolic (Ca 2+ ) levels to limit ASM cell CXCL10 production, reduced CXCL10 mRNA levels in histamine-treated cells would have been observed in this study but they were not. The lack of effect of the H1 and H2 receptor antagonists was surprising in view of our previous findings. Firstly, chlorpheniramine prevented histamine-induced changes in ASM cell GM-CSF and RANTES release following stimulation by IL-1 or TNF-. Secondly, the long acting 2 -adrenoceptor agonist salmeterol which, like histamine, increases cAMP levels also inhibited ASM cell CXCL10 release induced by these cytokines. Further, the effects of histamine appeared to be posttranscriptional, as cytokineinduced CXCL10 gene expression was not affected. Posttranscriptional effects of histamine on keratinocytes have been reported but were mediated via the H1 receptor. To our knowledge, H3 receptors are expressed on cholinergic nerves, but not ASM in the airways. Gantner and colleagues used specific primers for receptors H1-4 and detected mRNA expression for H1, H2, and H4 receptors, but not for H3 receptors. Perhaps the indirect inhibitory effects of histamine observed in this study were mediated via the Gi/o coupled H4 receptors. H4 receptors play an important role in the regulation of immune responses, including chemokine production, in the skin and so might also regulate ASM cell chemokine production. Further studies of cytokine-stimulated ASM cell CXL10 production in the presence of histamine are needed to determine the molecular mechanisms underlying its effects and histamine receptor involvement. Recently developed selective agonists (e.g., 4methylhistamine) and antagonists (e.g., JNJ7777120) for H4 receptors would be useful to clarify its role. ASM cells also express the organic cation transporter 3 (OCT3 or EMT) in addition to histamine receptors. Histamine is a good substrate for OCT3 and so it is possible that histamine entered the ASM cells stimulated with IL-1 and TNF-via OCT 3 and affected posttranscriptional events involved in CXCL10 protein synthesis/release. Further studies are needed to investigate the above possibilities and the mechanisms involved in any effects of histamine. Importantly, this study is the first demonstration that CXCL10 is susceptible to digestion by serine proteases such as tryptase. The levels of CXCL10 released following cytokine stimulation were reduced by tryptase, which is stored in the granules as a tetrameric complex with proteoglycans such as heparin and released at the same time as histamine. Unlike histamine, tryptase reduced detectable CXCL10 levels irrespective of the individual cytokine used to stimulate the ASM cells. This effect was due to tryptase protease activity as it was prevented by heat-inactivation and the serine protease inhibitor leupeptin. Further it was likely due, at least in part, to direct tryptase proteolysis of CXCL10, as tryptase reduced the levels of CXCL10 detected in a cellfree system. In addition, leupeptin added 3 h after any of the cytokines, when there is already a substantial increase in CXCL10 mRNA expression, completely prevented the reduction in cytokine-induced CXCL10 levels. These findings add CXCL10 to the chemokines CCL11 (eotaxin) and CCL5 (RANTES) that are also produced by ASM cells and cleaved by tryptase in vitro or by ASM cell cocullture with MC. Although other studies have investigated the effect of MC products individually on ASM cell synthetic function as above, this is the first study to also examine the overall effects of MC products released at different times after MC activation on asthmatic and non-asthmatic ASM cell CXCL10 production. As ASM cells do not release CXCL10 constitutively, or in response to FBS, IFN was used to induce CXCL10 release so the effects of the MC products on it could be assessed. The effects of rapidly released human lung MC products (2 h SN) on ASM cell CXCL10 levels complement the effects obtained with tryptase. Early MC products reduced asthmatic and non-asthmatic ASM cell IFN -induced CXCL10 levels by a similar amount. These effects were also prevented by leupeptin added later, indicating that serine proteases were involved, but do not affect early CXCL10 gene transcription. As well, a role for histamine cannot be completely dismissed. MC supernatants generated in the manner used here contain significant amounts of histamine, which would have been ineffective against the IFN -induced CXCL10 production used in this study. Early release of histamine Journal of Allergy may limit CXCL10 production generated under different inflammatory conditions when IFN is absent. In contrast to the effects of the MC products released early, newly synthesised products released sometime later (24 h SN) increased IFN -induced CXCL10 levels. Again the proportional effects of the MC products on ASM cells from people with and without asthma were similar. MC produce TNF-, which synergistically increases IFN -induced CXCL10 production by ASM cells in vitro. This study has extended those findings by providing evidence that products released from human lung MC activated by IgE also increase CXCL10 release and include sufficient TNFto cause such an increase. Although TNF-levels in MC SN collected early were half those in the SN collected later, they were still sufficient to amplify IFN -induced CXCL10 release. However no increase was observed, probably because the granule-derived proteases present in the SN cleaved the CXCL10 after it was released and thus reduced the amount of protein detected. In this study, the effects of MC products released together, either early or later, after IgE receptor activation were examined in vitro on ASM cells obtained from people with and without asthma. The MC could only be isolated from larger lung samples as they became available and were all from people without asthma. The effects of MC from people with asthma on asthmatic ASM cell CXCL10 production may be different. However different percentages of the MC SN were used to reflect what occurs in vivo, where activated MC are present in the ASM layer in different numbers in asthma. Thus not all ASM cells are in direct contact with a MC, but they have been observed in close proximity to each other. Given this proximity, it is apparent from our study that MC products could still affect ASM chemokine production in the absence of cell-cell contact. Conclusions This is the first study to examine the effects of MC products on CXCL10 production by ASM cells. Activated MC numbers are increased in the ASM in eosinophilic asthma and Tlymphocytes are present in the ASM in severe asthma and following allergen challenge. ASM-derived CXCL10 production may play a role in the recruitment of these cells to the ASM as both cell types express the CXCL10 receptor receptor CXCR3. In this study we have demonstrated that MC products released from granules soon after cell activation, or newly synthesized and then released later, differentially modulated levels of CXCL10 released by ASM cells from donors with and without asthma. Thus mast cells in the ASM have the potential to up-and downregulate chemokine levels locally and thereby affect their own recruitment and that of other inflammatory cells to the ASM in asthma.

A mixed model for repeated dilution assays. We propose a generalized linear mixed model to estimate and test marginal effects on titers repeatedly measured by serial dilution assays. The link is log-log and the titer is assumed to follow a gamma distribution. The parameters are estimated by generalized estimating equations. The marginal effects are tested by means of Wald and score tests, using the robust estimator of the variance. This approach avoids the problems arising from assays leading to nonestimable individual titers. Simulations were used to compare the Wald and score tests to Wilcoxon and Student t-tests. This method is applied to the comparison of the antiviral efficiency of three treatments against HIV.

Plasma surface functionalization induces nanostructuring and nitrogen-doping in carbon cloth with enhanced energy storage performance A facile, one-step and environmentally-friendly strategy for the preparation of hierarchical nitrogen-doped carbon cloth (hNCC) is presented via nitrogen plasma processing of commercial carbon cloth. In addition to N-doping, the RF plasma treatment induces nanostructuring, thus significantly increasing the surface area and liquid electrolyte wettability. The untreated carbon cloth (CC) has negligible Li-ion and supercapacitive storage capacities. However, after plasma treatment, the obtained hNCC delivers dramatically enlarged capacities. Specifically, the enhancement is by three times for Li-ion storage (150 mA h g−1versus 50 mA h g−1 at 100 mA g−1), and by three orders of magnitude for pseudocapacitance (391 mF cm−2versus 0.12 mF cm−2 at 4 mA cm−2). The effect of power-dependent plasma treatment for optimized performance is also investigated. We propose a plausible mechanism for achieving a hNCC architecture with highly enhanced ion/charge storage properties. Our research provides an approach to fabricate N-doped carbon materials with a controllable surface morphology and electrochemical properties. This deterministic and plasma-based method of preparing hNCC may offer new opportunities in the design and fabrication of high-performance carbon-based electrodes for energy storage devices.

“I have drafted legislation to help states and communities keep schools safe. Safe schools are for the most part a local responsibility, but in this case, the federal government can and should help." Sen. Lamar Alexander (R-Tenn.), as chairman of the Senate Education and Health Committee and sponsor of the Every Student Succeeds Act of 2015 and 21st Century Cures Act of 2016, has put together legislation, called the School Safety & Mental Health Services Improvement Act, to help improve student safety at our nation’s schools. “I have drafted legislation to help states and communities keep schools safe. Safe schools are for the most part a local responsibility, but in this case, the federal government can and should help. One way to do this is to give states and local school boards the authority to use federal dollars in different ways to make schools safer. For example, this bill would allow Tennessee to use federal dollars to upgrade school safety procedures, hire school counselors and develop mental health programs,” Sen. Alexander said in a press release. The School Safety and Mental Health Services Improvement Act would expand the use of federal funding to improve school safety, ensure federal mental health funds are made available to children and youth, and create a Presidential Task Force to better coordinate resources between the Departments of Education, Health and Human Services, Justice, Interior and Homeland Security. Alexander intends to introduce the legislation as early as Tuesday.

Tracking Stopping Times Let {(Xi, Yi)}i=1 be a sequence of pairs of random variables, and let S be a bounded stopping time with respect to {Xi}i=1. We propose the problem of finding a stopping time T with respect to {Yi}i=1 that optimally tracks S in the sense that T minimizes the average reaction time E(T − S) while keeping the false-alarm probability P(T < S) below a given threshold. This problem has applications in many different areas. In this paper we present an application related to communication over a channel with noisy feedback.

The present invention relates generally to cache management, and more particularly to maintaining cache coherence in a multiprocessor system. Advances in semiconductor fabrication technology have given rise to considerable increases in microprocessor clock speeds. Although the same advances have also resulted in improvements in memory density and access times, there remains a disparity between microprocessor clock speeds and memory access times. To reduce latency, often one or more levels of high-speed cache memory are used to hold a subset of the data or instructions that are stored in the main memory. A number of techniques have been developed to increase the likelihood that the data/instructions held in the cache are repeatedly used by the microprocessor. To improve performance at any given operating frequency, microprocessors with multiple cores that execute instructions in parallel have been developed. The cores may be integrated within the same semiconductor die, or may be formed on different semiconductor dies coupled to one another within a package, or a combination of the two. Each core may include its own level 1 cache which can store program data as well as instruction data (alternatively referred to as “instructions”) and an optional level 2 cache.

. Our study involving healthy postmenopausal females established that mammographic breast tissue density was lower in cases of more intensive stimulation by glucose of reactive insulinemia and glucose-induced glyoxalase I activity in bood mononuclears as well as in women with higher concentrations of circulating CD90+stem cells. Conversely, the density tended to increase in those with higher ratio of glucose-induced generation of reactive oxygen species in mononuclears. Our data point to possible mechanisms of increased density as a breast cancer factor when concomitant with relative predominance of progenotoxic effect of glucose and lower CD90+stem cells levels which are believed by some authors to be capable of suppressing the growth of certain tumors.

Conventionally, a use of a white light source, which is obtained by exciting a phosphor by using excitation light of semiconductor laser or the like, as a light source for projection, which is included in a headlamp of a vehicle, has been proposed. As illustrated in FIG. 11, in accordance with a light distribution characteristic standard which is specified by laws, a headlamp of an automobile Car is required to be configured to form a light distribution pattern BP which has linear and sharp light-dark contrast CL in a front side of the automobile Car, for example, in a case where the headlamp is a lamp for passing. Such light-dark contrast is referred to as cutoff. As one example of the headlamp, PTL 1 describes a lamp for a vehicle, which includes a light emitting section composed of an LED light source which forms a light distribution pattern for a lamp for passing and a phosphor for a laser light source which radiates fluorescence to a vicinity of a cutoff having the aforementioned light distribution pattern. By arranging a shade having the same shape as that of a cutoff line in close proximity to or immediately above a light-emitting surface of the phosphor, the lamp for a vehicle obtains a light distribution pattern having a shape prescribed with the light distribution characteristic standard for a headlamp.

The Effectiveness of Coursework and Onsite Coaching at Improving the Quality of Care in InfantToddler Settings Research Findings: This study evaluated the effectiveness of 2 professional development interventions aimed at improving the quality of care provided by caregivers in ordinary infanttoddler child care settings, both center- and home-based. In all, 183 participants in a community college course on infanttoddler theory and practice, an in-service community model of a 48-hr course plus various levels of 1-on-1 coaching (participants were randomly assigned to 0, 5, or 15 hr), or a no-intervention comparison group were compared on their changes across 3 time points in their job-related self-efficacy, their knowledge of best practices, and the quality of their interactions with children as assessed by objective observers. Results indicated that the group with the most intensive intervention (course plus 15 hr of coaching) displayed the most consistent pattern of improvements, which met multiple analytical criteria for substantiveness. Furthermore, these findings were most apparent in the quality of interactions outcomes, and these were further still concentrated in the area of support for language and learning, the domain that showed the greatest need for improvement in infanttoddler caregivers in this sample as well as in previous research. Practice or Policy: Implications of the study results for systemic improvements to infanttoddler care are discussed, such as the importance of individual-level professional development, minimum dosage of coaching, and in-service supports to prevent declines in the quality of teacherchild interactions.

But for most people, the time spent juggling two windows or scrolling across large documents isn’t the biggest bottleneck in getting work done. Instead, there’s a more basic, pernicious reason you feel constantly behind — you’re getting distracted. Ms. Mark’s research, based on observations and digital tracking of office workers, has found that our workplaces are bombarded with distractions. Studies show that office workers are interrupted every four to 11 minutes by external distractions including phone calls, email and people who stop by your desk to chat about the weekend. Then there are self-motivated distractions, when, for no apparent reason, you quit working on your project and do something else — for instance, jump into the rabbit hole of the web. All such disruptions are costly. It can take workers as much as 25 minutes to regain focus after being interrupted. And constant interruptions create a stressful workplace. “The second screen can also be an inviting entry-way for self-distraction,” Ms. Mark said. That’s because it’s an ever-present, available canvas calling out for you to fire up a web window and find solace in the latest thrills on YouTube. Adherents to the multimonitor lifestyle might argue that the second screen allows people to deal with their distractions more quickly. To check your email on a single-screen machine, you may find yourself frequently switching from your main task to your inbox and back again. With two monitors, you can always keep an eye on the inbox. This feels efficient; dedicating a swath of screen space to your favorite distraction feels as if you’re somehow managing your habit. But if you wouldn’t watch a movie or play a video game while you’re trying to get something done, why would you keep an app as distracting as email sitting within your field of view? Like many researchers who study attention, Ms. Mark recommends that people create work habits that reduce the chance of interruption. For instance, she suggests that people turn off email notifications, and answer and write email in batches once or twice a day rather than every few minutes. She notes that taking up such habits requires both personal discipline and buy-in from your bosses and co-workers. That gets to the blessing of one monitor. With a single screen, I was forced to fight my distractions. I had to actively prevent myself from falling into email and Twitter, from ever losing focus on my main window. It took some time for me to exercise that willpower. But by finding methods of sticking to my task rather than coping with my distractions, my single-screen machine ultimately improved how I work. It can for you, too — if only you resist the pull of two displays.

Project Management in Information System Development Education We have taught a laboratory course on system development at the School of Information Science of Kyushu Sangyo University since 2004. Based on our experience, we decided to address various problems of the education in information system development. We categorized these problems from three viewpoints. First, from the viewpoint of system development, the specific project and the method of development are important. Second, from the viewpoint of the project laboratory, it is difficult for students to do their projects effectively. Third, from the viewpoint of project management, it is a problem for both teachers and students to follow the progress of a project and to maintain control of it. In 2007, in order to solve these problems, we made several improvements, including adding Personal Software Process and Earned Value Management.

Psychological and neuropsychological correlates of dependence-related behaviour in Medication Overuse Headaches: a one year follow-up study Background Medication Overuse Headache (MOH) can be related in some patients to dependence-related behaviour characterised by craving, a deficit in controlling substance intake, which is associated to orbitofrontal cortex (OFC) dysfunction. The aim of this study was to explore the psychological correlates in MOH patients and the functioning of the OFC through neuropsychological assessment (Iowa Gambling Task: IGT) and to relate it to prognosis at a one year follow-up point. Findings Seventeen subjects suffering from probable MOH were included and compared to 19 migraineurs and to 17 controls. The results show significant between group differences for behavioural dependence, depression, anxiety, catastrophizing. There were no between group differences for impulsivity. Mean IGT score did not allow differentiation of MOH patients from the other groups, whereas the score was significantly different between opiate abusers and other medication abusers (45 +/−5.7 versus 57.1 +/−8.2, p=0.019). Among the clinical variables rated at inclusion, the amount of acute headache medication taken per month was the only one predicting the prognosis (RR=1.05, 95% CI=1-1.06, p=0.04). A slight increase in risk of relapse at 1 year was observed in patients with poorer IGT scores (RR=0.92, 95% CI=0.85-1, p=0.05) and higher behavioural-dependence scores (RR=1.07, 95% CI=11.14, p=0.05). None of the other psychological variables predicted relapse risk. Conclusions These results must be interpreted with caution due to the low number of subjects. They showed a deficit in decision making processes in MOH patients who overuse medications containing psychoactive substances like opiates. Moreover dependence-related variables are related to the prognosis. Introduction The role of behavioural disorders and particularly an addictive component of behaviour in Medication Overuse Headaches (MOH) is a matter of debate. Orbitofrontal dysfunction is known to be at work in addictive behaviour. Indeed, orbitofrontal cortex is involved in the ability to inhibit craving behaviour through decision making impairment. Other psychological dimensions may be implicated in dependence related behaviour in MOH patients. In particular, MOH patients have been shown to be at risk of depressive disorders and anxiety disorders when compared to episodic migraine patients. Cupini and al. emphasized the presence of obsessive compulsive disorders in these patients as part of the compulsive drug-seeking behaviour. In a one year follow-up study anxiety and depression were not reliable prognosis factors for MOH, in contrast to addictive behaviour. Nevertheless, no study has focused on catastrophizing in MOH patients or on impulsivity while those dimensions may be of interest in determining dependence related behaviour. Our objective was therefore to compare anxiety, depression, catastrophizing and impulsivity dyscontrol as well as decision-making impairment in MOH patients, episodic migraine patients and healthy controls. Then, a follow up was performed in order to relate psychological and neuropsychological variables to prognosis. Methods This is the clinical part of an imaging study implying rest fMRI and 18 F FDG PET. Results of the imaging study will be presented elsewhere. The study was prospective, non-randomised, comparing MOH evolving from migraine patients to episodic migraine sufferers (EM) and to healthy volunteers without headache (HV). MOH and EM patients have been followed up over one year. Patients asking for a consultation at the neurological department at the University Hospital in Bordeaux were included if they fulfilled migraine diagnostic criteria (ICHD-II 1.1 and 1.2) or probable MOH diagnostic criteria (ICHD-II 8.2) with prior migraine and if they were 18 years old or more. A prior episode of MOH was an exclusion criteria for the migraine group The healthy volunteers were recruited among Bordeaux University Hospital staff members. Clinical data collection comprised a standardized interview process inquiring for crisis frequency and medication intake frequency during the last three months and acute and prophylactic antimigraine drugs currently used. Patients from the EM and MOH groups were given a headache diary, recording days with headache and acute headache medication intake during the follow-up period. All subjects answered self-administered questionnaires. In order to assess depression, the Beck Depression Inventory ((BDI-13 items) was used. Anxiety was assessed using the State Trait Anxiety Inventory (STAI-state). Catastrophizing was assessed using the Pain Catastrophizing Scale (PCS). The Barratt Impulsiveness Scale (BIS-11) is a self-report measure of impulsive personality traits. The Medication Dependence Questionnaire for Headache sufferers (MDQ-H) is a rating scale assessing dependence to medication in headache patients. Then, all subjects underwent IGT, a computerised task which is a test aiming to detect decision-making impairment. It is a test where the subject is confronted with a decision that involves a conflict between an immediate reward and a long term negative consequence. The lower the score achieved, the poorer the decision-making performance is. Subjects from the MOH group underwent either an inpatient or outpatient withdrawal procedure. MOH and EM groups were re-examined at 12 months from inclusion. Patients from the MOH group were classified in a binary way as relapse or non relapse depending on whether they fulfilled ICHD-II criteria for MOH at month twelve visit. Statistical analysis was performed using SPSS version 17.0 software. Descriptive analysis was performed and the differences between groups were tested using chi square and a one-way analysis of variance when appropriate. In order to compare the score results obtained by the questionnaires and the IGT between the three groups a one-way analysis of variance was also used, and t-test was used to compare groups in pairs. Logistic regression analysis was performed in order to predict relapse within one year. For the patients that have been followed up during one year, scores evolution was tested using paired t-test. Findings Fifty-one subjects were included in the study: 17 patients suffering from MOH, 19 EM 17 HV. Sociodemographic characteristics of participants are presented in Table 1.The three groups were comparable for sex, age, family situation and work status. The mean number of days with headaches was 23.5 +/− 7.1 in the MOH group and 2.5 +/−1.1 in the EM group. The mean duration of MOH was 6.8 years; duration ranged from 4 months to 30 years. The acute headache medication overused by MOH patients are shown in Table 2. Five patients were overusing either opiates or a combination of analgesics containing opiates derivates. The total amount of acute headache medication taken per month was 82 in the MOH group and 6.2 in the EM group. The mean scores obtained in each questionnaire assessing the psychological dimension in each groups are presented in Table 3. The results show that mean scores were higher in MOH than in EM and HV for dependence (MDQ-H), depression (BDI), anxiety (STAI), catastrophizing (PCS). The between groups analysis of variance for anxiety (STAI)) and depression (BDI) indicated a significant statistical group effect (F = 5,7, p = 0;006 ; F = 6;3, p = 0;003). On the contrary, there were no between group differences for impulsivity (BIS). Total raw scores for IGT did not allow to differentiate MOH patients (IGT total raw score score = 26.2+/−24.8) (Table 2). Among the clinical variables rated at inclusion, the number of acute headache medication units taken per month was the only one predicting the prognosis (RR = 1.05, 95% CI = 1-1.06, p = 0.04). A slight increase in risk of relapse at 1 year was observed in patients with poorer IGT raw scores (RR = 0.92, 95% CI = 0.85-1, p = 0.05) and higher MDQ-H scores (RR = 1.07, 95% CI = 1-1.14, p = 0.05). None of the other psychological variables predicted relapse risk. For the MOH group a one year a significant decrease was observed for MDQ-H (t = 5,7, p < 0,000) and for PCS (t = 3,7, p = 0,003) and for the EM groups no significant differences were observed. Discussion To summarise, this study showed between group differences for anxiety, depression, catastrophizing and behavioural dependence but not for impulse dyscontrol and decision-making impairment. However, MOH with opioid medication overuse exhibited a significant disturbance in decision making when compared with MOH overusing other acute headache medication. Moreover dependence related behaviour and decision-making impairment at inclusion weakly predicted one year prognosis in MOH whereas other variables did not. Catastrophizing has been reported to be associated with a higher risk of prescription opioid abuse in pain patients. In the field of headaches, it has been showed that subjects with chronic daily headaches exhibit increased catastrophizing scores when compared to episodic migraineurs, higher catastrophizing being associated with lower quality of life. Moreover the prevention of MOH in high frequency migraine patients by a cognitive behavioural program is associated with a lowering of catastrophizing. There are a few studies examining psychological factors as predictors of prognosis in MOH patients. The Akershus study of chronic headache showed that Dependence scores (SDS) predict 2 to 3 year prognosis after detoxification. Smoking, alcohol consumption and the doses of antimigraine drugs used allowed to predict one year follow-up relapse in detoxified MOH but not in anxiety disorders or affective disorders. In a one year follow-up study of 72 subjects with MOH from the general population Fontanillas showed that opioid abuse was associated with a bad prognosis. In this direction, our results confirm that the dependence related behaviour is a stronger predictor of prognosis than emotional disturbances. Indeed, despite the small size of the sample, the amount of acute headache medication taken and the MDQ-H score at inclusion significantly predicted prognosis at one year whereas all emotional measures did not. The IGT allows to measure the ability to defer immediate reward, taking into account the long term negative consequences, and it is altered in prefrontal-damaged patients. In a sample of MOH patients, Biagianti and al showed a statistically significant deficit in IGT net score compared to healthy matched controls. Patients were retested one year later and IGT performance remained negative. The IGT score did not allow to predict relapse in medication overuse after detoxification. In a similar vein, Gomez-Beldarrain evaluated orbitofrontal impairment using other neuropsychological tasks. They found a significant impairment in the orbitofrontal tasks performance in MOH compared with controls. Moreover, in the one year follow-up this performance predicted relapse in MOH. Our study failed to demonstrate any difference between MOH and the two controls groups despite a slightly lower performance in MOH than in episodic migraineurs. Our healthy control sample clearly exhibited a lack of motivation when performing the task. On the other hand, the comparison between opiate overusers and other medication overusers showed a significant difference between the two groups on the IGT total net score. This result can be put into parallel with the fact that in Fumal's sample of MOH patients, the hypofunctioning of the orbito-frontal cortex showed with PET was due to the subsample of patients overusing opiates. Among the MOH sample, the IGT total net score predicted the prognosis at a one year follow up visit confirming the previously cited studies. The strength of this study is its one year prospective design, the assessment of rarely explored psychological dimensions such as impulsivity and catastrophizing. The major weaknesses are the small number of subjects and the motivation failure in the healthy controls, thereby it would be very usefull to present further resuklts with more subjects. Nevertheless the study allows to confirm the utmost importance of dependence related behavioural disturbance in characterising MOH patients. Moreover it seems determinant for prognosis in an illness characterised by its poor prognosis. These results lead us to advise strong management of the psychological dimension in MOH patients. Cognitive behavioural therapies in particular can address catastrophizing, emotional disorders and impulse dyscontrol and motivational interviewing should be part of the management of the dependence related behaviours. Authors' contribution FR: conception and design, acquisition of data, statistical analysis, interpretation of data and discussion of results, drafting the article, final approval of the complete manuscript. SC: interpretation of data and discussion of results, final approval of the complete manuscript. G DS: interpretation of data and discussion of results, final approval of the complete manuscript. VD: acquisition of data, discussion of results, final approval of the complete manuscript.MA: conception and design, interpretation of data and discussion of results, final approval of the complete manuscript. All authors read and approved the final manuscript. Financial support This study has been granted by Institut UPSA de la Douleur.

Research Of Physical And Mechanical Indicators Of Jensie And Knitted Fabrics Recommended For Children's Combined Outerwear The article examines and analyzes the physical and mechanical properties of different proportions of jeans and knitted fabrics recommended for children's combined outerwear. It also offers the most alternative fabric patterns for children's outerwear. INTRODUCTION In today's world of clothing, not only the consumer demand or fashion trend, but also the right choice of fabric to suit the product, depending on who and for what purpose, for what season, at the same time, special attention is paid to creating a model that matches the characteristics of the fabric. OCLC -1091588944 processes of production, labor productivity, and product quality. By designing high-quality children's sportswear and correctly approaching the modern economic features of their production, production as a high quality seasonal sewing item. The practical and scientific design of clothing plays a leading role in solving the problems of the garment industry, because at this stage of design, the social and technical-economic requirements for clothing and its quality must be fully taken into account. Based on these requirements, a new clothing model will be developed. In children's sportswear, first of all, the fabric chosen for sewing clothes should be resistant to the dynamic movements of children, which should serve to improve air circulation in them. In a market economy, the demand for knitwear is growing day by day. One of the main requirements of today is to expand the range of top knitwear for children, given the current technological capabilities. It is known that different types of knitted fabrics are produced: glad, fleece, chain, knitwear, sukno, sharme, and others. Due to the high natural properties of the study -cotton, linen, wool -fibrous fabrics were selected as the preferred fabrics for children's combined clothing, knitwear of different sizes and denim fabric were selected. In order to determine the physical and mechanical properties of knitted fabrics from 10% lycra with 90% cotton fiber, 7% lycra with 93% cotton fiber, 5% lycra with 95% cotton fiber, as well as from 50-60 tex warp and weft yarns, samples with a mass of 282-340 g of 1 m2 of fabric woven in twill weaving were performed on modern equipment installed in the testing laboratory of the Namangan Institute of Engineering and Technology in order to determine the physical and mechanical properties of the selected fabrics. Sampling of knitted fabrics was carried out in accordance with GOST 8844-75. Using the DW-1111 machine, the samples were cut to 100x100 mm and the weight of the fabric was measured using an ZK-200C electronic vacuum scale ( It is known that denim are made of cotton yarn or yarn mixed with polyester. Denim fabrics belong to the group of garment fabrics, which are woven from combed yarn and carded yarn spun with a relative linear density of 60 to 100 tex and above, and the mass of 1 m2 of fabric is 250-300 g. will be. Clothing is in direct contact with the body, and its quality is taken into account in the design of the product. Therefore, it is necessary to determine the fiber content and physical properties of the fabric. The air permeability of fabrics is one of the hygienic indicators for children's outerwear, determined by the coefficient indicating the amount of air passing through 1 cm 2 of fabric per second from the pressure difference applied on both sides of the fabric. Air permeability on the device YG-461E in accordance with the standard B (cm 3 /cm 2 sec) GB / 5453 (ISO 9237) for ready-made garments -pressure -100Pa, range size -.08.0 mm under normal conditions tested and determined using the following formula. Here: V -the amount of air passing through the fabric at a given pressure difference , cm 3 ; S -fabric area,cm cm 2 ; T -is the time taken for the air to pass through the fabric, sec. The air permeability in the selected knitted fabrics varied from 44,865 cm 3 / cm 2 se to 58,051 cm3 / cm 2 sec (Fig. 2) The main quality indicator of fabrics is characterized by tensile strength. In the research work, the breaking force was prepared according to the standard method, the length and width of the samples were 30 x 5 cm, and for 30 minutes using a dynamometer "YG-026T" -454 x 3 (13 N). results were obtained. The results obtained were calculated using the following formula ( In the analysis, the re-deformation (height) of the sample varied from 0.07% to 97.3%. The lowest result was observed in variant-II knitted fabric (Fig. 4). The relative elongation in the selected jeans fabrics ranged from 10.50% to 38.95%. The lowest relative elongation was found in variant 1 fabric. In denim fabrics, the lowest air permeability from 3,751 cm 3 / cm 2 *sec to 5,260 cm 3 / cm 2 *sec was observed in the selected variant 1 of Denim fabric ( Table 2). The relative elongation ranged from 10.50% to 38.95%. The lowest relative elongation was found in variant 1 fabric. The results show that the knitwear patterns have significantly higher elongation than Denim fabrics and have changed significantly even under the influence of a small force. The results obtained from the selected fabric samples showed that the re-deformation Another factor to consider when choosing a fabric for children's outerwear is the abrasion resistance of the fabric. Fabric samples were tested using the YG401B friction tester. Fabric samples of 100x100 mm were subjected to a load of 9 kPa at a speed of 20-70 rpm and placed at 90,000 cycles. The degree of peeling of fabrics was observed from 859 to 2692 cycles when the equipment was started at a speed of 70 rpm under a load of 9 kPa. The results show that the highest peeling fabric is in the variant III fabric, low-peeling fabric was observed at 2692 cycles in variant I fabric. Fabric degradation was the highest in the sample, with 19,332 cycles degraded in variant II fabric. The most durable non-abrasive fabric was observed in these I and V variants. The results are given in Table 3. CONCLUSION Research shows that it is very important to choose a suitable fabric for children's outerwear, taking into account not only their aesthetic but also their physical and mechanical properties.

. INTRODUCTION Little data exist about the prevalence of psycotherapy used by the general population of the city of Buenos Aires. The aim of our study was to determine this prevalence, the demographic characteristics of the population that receives psychotherapy, as well as types and characteristics of these psycotherapies. METHOD A survey was conducted among 1510 inhabitants in different neighbourhoods of the city of Buenos Aires. RESULTS During the month previous to the survey, 15.6% of the population received psychotherapy treatment, 21% during the last year and 41.6% had received before. Psychoterapy treatment is more prevalent among women and people with higher educational and social-economic level, and it is less frequent in the elderly group. No differences in the use of psychotherapy exist according to marital status. Forty four percent of the patients under treatment at the moment of the survey were unaware of the type of psychotherapy received, 41.3% knew that they were receiving psychoanalytic therapy and 8% cognitive-behavioral psychotherapy. Eighty-two percent of the treatments were conducted by psychologists and 11% by physicians. CONCLUSIONS The use of the psychotherapy is very extended among the general population of the city of Buenos Aires.

Into Ones Own, or on Finding Oneself This chapter takes up the popular notion of finding oneself, thematized in literature from the nineteenth-century Bildungsroman to the fiction of Jack Kerouac of the beat generation and the poetry of Robert Frost. It considers Socratess call to know thyself in light of existentialist criticism. It considers especially Sartres criticism of the idea of the self as an inner core or essence that determines who we are, and that could be thought to be found in self-seeking. It examines Nietzsches notion of self-becoming and the phenomenological rendering of the self as transcendence toward possibility.

Growth and endotoxin production of Yersinia enterocolitica and Enterobacter agglomerans in packed erythrocytes Since 1987, the Centers for Disease Control investigated six cases of transfusion-associated sepsis. All six patients developed septic shock after receiving units of packed erythrocytes (PRBCs) contaminated with Yersinia enterocolitica (five patients) and Enterobacter agglomerans (one patient); three of the blood recipients died. We studied the growth and endotoxin production of Y. enterocolitica and E. agglomerans in units of PRBCs stored at 4 degrees C for 60 days. When PRBCs were inoculated with 0.1 to 1.0 CFU of these organisms per ml, both Y. enterocolitica and E. agglomerans entered log-phase growth 2 to 3 weeks after inoculation; generation times were 15 and 22 h, respectively. Endotoxin was first detected at 3 weeks following inoculation, and the concentration paralleled the log phase of growth of the strains tested. These data show that prolonged storage of PRBCs at 4 degrees C provides conditions that allow these two organisms to grow and subsequently produce high concentrations of endotoxin.

New scientific diagnostics in the ESMValTool an overview <p>The Earth System Model Evaluation Tool (ESMValTool) has been developed with the aim of taking model evaluation to the next level by facilitating analysis of many different ESM components, providing well-documented source code and scientific background of implemented diagnostics and metrics and allowing for traceability and reproducibility of results (provenance). This has been made possible by a lively and growing development community continuously improving the tool supported by multiple national and European projects. The latest major release (v2.0) of the ESMValTool has been officially introduced in August 2020 as a large community effort, and since then several additional smaller releases have followed.</p><p>The diagnostic part of the ESMValTool includes a large collection of standard &#8220;recipes&#8221; for reproducing peer-reviewed analyses of many variables across ESM compartments including atmosphere, ocean, and land domains, with diagnostics and performance metrics focusing on the mean-state, trends, variability and important processes, phenomena, as well as emergent constraints. While most of the diagnostics use observational data sets (in particular satellite and ground-based observations) or reanalysis products for model evaluation some are also based on model-to-model comparisons. This presentation gives an overview on the latest scientific diagnostics and metrics added during the last year including examples of applications of these diagnostics to CMIP6 model data.</p>

Use of non-orthodox and conventional health care in Great Britain. OBJECTIVE--To describe the characteristics of patients using non-orthodox health care and their pattern of use of conventional health care with respect to a particular problem. DESIGN--Postal survey of all 2152 practitioners of acupuncture, chiropractic, homeopathy, naturopathy, and osteopathy identified from 11 national professional association registers. Patients attending a representative sample of 101 responding practitioners completed questionnaires covering demographic characteristics, presenting problems, and use of the health service. SETTING--Practices of practitioners of non-orthodox health care in England, Scotland, and Wales. SUBJECTS--Qualified, non-medical practitioners of non-orthodox health care working in Great Britain and 2473 patients who had attended one of the sampled practitioners in an allocated time period between August 1987 and July 1988. RESULTS--An estimated 1909 practitioners were actively practising one of the study treatments in Great Britain in 1987. Of the estimated 70,600 patients seen by this group of practitioners in an average week, most (78%) were attending with a musculoskeletal problem. Two thirds of the patients were women. Only 2% were aged under 16, but 15% were aged 65 or over. One in three patients had not received previous conventional care for their main problem; 18% were receiving concurrent non-orthodox and conventional care. Twenty two per cent of the patients reported having seen their general practitioner for any reason in the two weeks before the surveyed consultation. CONCLUSIONS--Patients of non-orthodox health care, as provided by this group of practitioners, had not turned their backs on conventional health care. Non-orthodox treatment was sought for a limited range of problems and used most frequently as a supplement to orthodox medicine.

Parasutterella secunda sp. nov., isolated from human faeces and proposal of Sutterellaceae fam. nov. in the order Burkholderiales. A novel, strictly anaerobic, non-spore-forming, Gram-reaction-negative coccobacillus bacterium, designated strain YIT 12071(T), was isolated from human faeces. Biochemically, this strain was largely unreactive and asaccharolytic. Growth of this strain in peptone-yeast-extract broth was weak, producing no visible turbidity, and no short-chain fatty acids were detected as an end product of metabolism. Following 16S rRNA gene sequence analysis, strain YIT 12071(T) was found to be most closely related to Parasutterella excrementihominis (90 % sequence similarity) and phylogenetically distinct from other known genera belonging to the order Burkholderiales. Biochemical data supported the affiliation of this strain with the genus Parasutterella. Strain YIT 12071(T), therefore, represents a novel species of the genus Parasutterella, for which the name Parasutterella secunda sp. nov. is proposed. The type strain is YIT 12071(T) (=DSM 22575(T) =JCM 16078(T)). On the basis of 16S rRNA gene sequence analysis, species of the genera Sutterella and Parasutterella form a distinct and deep evolutionary lineage of descent in the order Burkholderiales. This lineage could not be associated with any of the four known families of the order Burkholderiales. The distinct phylogenetic position and the unusual combination of chemotaxonomic characteristics shared by these genera, such as the predominant quinones and cellular fatty acid compositions, suggest that they constitute a novel family in the order Burkholderiales, for which the name Sutterellaceae fam. nov. is proposed to accommodate the genera Sutterella and Parasutterella.

Parents say a draft action plan to tackle racism, Islamophobia and concerns over foreign travel by trustees at the York Region District School Board is “insulting” and “ineffective” and unlikely to restore confidence in the troubled board. Last November, the Minister of Education sent the board a strongly worded letter demanding concrete steps to address community concerns, particularly around the board’s handling of issues of equity and discrimination. The board’s response is due by Friday. In a draft letter to be discussed at a special board meeting Wednesday evening, the board says it is hoping mandatory trustee sensitivity training, an equity audit of the organization and a commitment to being more transparent will restore faith in the board. “It is clear that better communication and understanding is key to resolving many of these issues — both with our communities and between staff and trustees,” says the letter, written by the new trustee chair Loralea Carruthers, with the input of other trustees. “It is also essential that we provide professional development for trustees and staff in areas that need addressing such as equity and the role of trustees in school board oversight. “Furthermore, examining our policies to ensure that they are clear, fair and being followed in the spirit they were intended, is important for transparency and accountability,” the letter says. But parents and observers say the letter is “insulting” and offers few real solutions or specifics about how future problems will be addressed — or clarity on how they have been handled in the past. For example, the letter describes confusion when it came to investigating anti-Muslim Facebook posts of a Markham elementary school principal. Parents who raised concerns were initially told the posts were being investigated under the Respectful Workplace and Learning Environment policy, which would have required the board to engage complainants and keep them informed about how the investigation was progressing. In the end, the matter was probed under a completely different policy, called Professional Misconduct and Progressive Discipline, that allowed the issue to be dealt with internally, the letter states. “Parents were not deliberately misled,” said Carruthers in an interview Monday, adding there was some confusion within the board as well. In an email, director J. Philip Parappally said, “There was one incident of a miscommunication between a parent and a staff member, however the Professional Misconduct and Progressive Discipline policy, which is the applicable policy, was used and all appropriate measures were taken,” he said. The principal, Ghada Sadaka, who later apologized for the posts, is still on a personal leave. Both policies do not allow the board to disclose the outcome of the investigation. “This is insulting to the parents. We got blocked from every way, and now they are saying miscommunication,” said Markham parent Naeem Siddiqi. “It’s an issue of doing a sham investigation and now they are trying to pin it on miscommunication,” he said. Among the actions trustees plan to take are: professional development on governance, and on racism and equity. The board is also discussing a third party audit of equity practices. “We want a very independent look at this by someone who has no connection with the board,” said Carruthers. But parents are not convinced. “No amount of audit, meetings, and training is going to fix the trust that has been lost,” said parent Charline Grant, who has a human rights complaint against the board for allegations of racism. “It’s not enough,” she said. “It appears to be a ‘quasi-dysfunctional’ environment, because if there are so many equity concerns raised, it should be a collaborative effort by all the players to address the concerns of parents, and staff,” he said. Parents say they are disappointed by the draft, and were expecting more as many are optimistic the recent election of Carruthers will usher in a period of change at the board. Carruthers, meanwhile, indicated the board is listening.

The present invention relates to system maintenance and diagnosis, and more particularly to techniques for non-intrusive gathering of diagnostic data in a monitored system. When a monitored system encounters a failure or error, diagnostic data is typically collected and stored to a disk for diagnostic analysis (also referred to as dumping diagnostic data to a disk). The diagnostic data may then be communicated to a diagnosis site for analysis and resolution of the error. The diagnostic data that is collected and stored (also referred to as diagnostic data dumps) can be quite large measuring in thousands of files and several gigabytes of data. As a result, the process of gathering and storing diagnostic data can be time consuming. This can have several negative impacts on the system. For example, when a process or thread encounters an error in the system, the failing process or thread that receives the error may be in a suspended state until the diagnostic data gathering has completed. As a result, an end user sending a request to the failing process or thread may receive no response from the failing process or thread for the request since the failing process or thread is in a suspended state. In addition, if the failing process or thread holds a lock on some system resources that are shared with other processes or sessions, these other processes or sessions may be blocked for the duration of diagnostic data gathering caused by the failing process or thread. Furthermore, due to the size of the diagnostic data being gathered and stored, the task of gathering and storing the diagnostic data can be very resource intensive and may negatively impact the system as a whole. The situation is further aggravated if the failures are encountered by a large number of processes or threads in the system causing the failing processes or threads to collectively exhaust system resources.

Low-dose Risedronate Sodium Protects Bone Cells after Abrupt Oestrogen Withdrawal Objective: To investigate the effects of low-dose risedronate sodium on the in vitro cellular profile of osteoblasts, adipocytes, osteocytes and osteoclasts in a rat model of abrupt oestrogen deficiency. Methods: Oestrogen deficiency was induced by ovariectomy in 24 female rats. The rats were treated with low-dose (0.24 g/kg) or high-dose (2.4 g/kg) risedronate sodium for 4 days presurgery, continuing every 3 days until 15 days postsurgery. Osteogenic and adipogenic differentiation were determined in cultured bone marrow cells by alkaline phosphatase and Oil Red O staining, respectively, and by osteogenic and adipogenic gene expression. Osteoclast formation was measured in bone marrow cells stimulated with macrophage colony-stimulating factor and receptor activator of nuclear factor kB ligand, and stained with tartrate-resistant acid phosphatase. Osteocyte apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling assay and B-cell lymphoma-2 (Bcl-2) immuno - histochemistry. Results: Low-dose risedronate sodium enhanced osteoblast differentiation, suppressed adipocyte differentiation and osteoclast formation, and reduced osteocyte apoptosis through regulation of Bcl-2 and Bcl-2-associated X protein. Conclusions: Low-dose risedronate sodium may have clinical benefit in protecting against bone loss after abrupt oestrogen deficiency.

What Drives the German Current Account? And How Does It Affect Other EU Member States? We estimate a three-country model using 1995-2013 data for Germany, the Rest of the Euro Area (REA) and the Rest of the World (ROW) to analyze the determinants of Germanys current account surplus after the launch of the Euro. The most important factors driving the German surplus were positive shocks to the German saving rate and to ROW demand for German exports, as well as German labour market reforms and other positive German aggregate supply shocks. The convergence of REA interest rates to German rates due to the creation of the Euro only had a modest effect on the German current account and on German real activity. The key shocks that drove the rise in the German current account tended to worsen the REA trade balance, but had a weak effect on REA real activity. Our analysis suggests these driving factors are likely to be slowly eroded, leading to a very gradual reduction of the German current account surplus. An expansion in German government consumption and investment would raise German GDP and reduce the current account surplus, but the effects on the surplus are likely to be weak.

Biosynthesis, characterization and antibacterial activity of silver nanoparticles using an endophytic fungal supernatant of Raphanus sativus Graphical abstract Introduction There is an increasing commercial demand for metal nanoparticles due to their potential applicability in various areas such as electronics, catalysis, energy, textile and medicine. Among various metal nanoparticles, silver nanoparticles (AgNPs) in particular have been the focus of increasing interest due to their peculiar properties which can be tailored for a specific application by controlling the shape, size and morphology of the nanoparticles. The available physical and chemical methods for metal nanoparticle synthesis resulted in low production rate, high expenditure and release of toxic chemicals. Moreover, the large-scale synthesis of metal nanomaterials suffers from certain drawbacks such as stability and polydispersity, especially if the reduction is carried out in aqueous media. Thus, biological synthesis of AgNPs provides a promising alternative offering advantages such as environment friendly, easily scaled up for large-scale synthesis and there is no need to use high pressure, temperature and toxic chemicals. Biological methods based on microbes such as fungi, bacteria and viruses are able to absorb and accumulate metals and can be used in the reduction and control of nanostructural topography of metal ions and thus used for the synthesis of nanoparticles. Endophytic fungus are more ideal for the synthesis of nanoparticles as the fungi could form large biomass which can withstand agitation and flow pressure and other conditions in bioreactors or other chambers and be compared to bacteria and plant materials. These are fastidious to grow, secrete large amount of enzymes and are easily handled in downstream processing and thus easy for fabrication of AgNPs and can be directly used in various applications. In pharmaceutical industry, AgNPs are the most promising as they show good antibacterial activity against an array of pathogens due to their large surface area to volume ratio. One such size-dependent property has led to the development of dressing materials incorporated with nano-sized silver to enhance the wound healing property of the dressing materials. The biosynthesized AgNPs have been applied widely to prevent biomedical devices associated infections, food preservation, water purification, clothing, cosmetics and other numerous pharmaceutical products. Endophytes are microorganisms that reside inside living internal tissue of plants without inflicting any immediate, overt negative effects to the host plants. Endophytes have mutualistic relationship to their host, often protecting plants against pathogens and in some cases they can act as opportunistic pathogens. Drug resistance in human pathogens is a big challenge in fields such as pharmaceutical and biomedicine. One such prime focus of the pharmaceutical sector is to develop potent antibacterial drugs to combat drug-resistant pathogens which are growing at alarming pace with limited choice of available antibiotics. At present, various conventional strategies using standard antibacterial agents have been used for the treatment and prevention of pathogenic bacteria. But, some of the bacterial strains have acquired resistance against available antibiotic therapies and pose the greatest risk of infection which necessitates the search for alternative drug or natural antibacterial. Therefore, modification or development in antimicrobial compounds to improve bactericidal potential is a priority area of research in the modern era. Thus, endophyte studies have been important to discover substances which will be utilized in a wide variety of disease causing agents. Endophytes may produce a plethora of substances having use in modern medicine, agriculture, antimycotics, immunosuppressant and anticancer compounds. To date, the endophytes have been most extensively studied for their ability to produce antibacterial, antioxidants, anticancer, antidiabetic, antiviral and antisuppresive compounds and their use in the biosynthesis of nanoparticles was also mentioned to some extent. Therefore, in the present study, endophytic fungi Alternaria sp. was isolated from healthy leaves of Raphanus sativus for biosynthesis of AgNPs. The synthesized AgNPs were characterized using UV-vis (UV-vis) spectroscopy, Fourier transform-infrared (FTIR) spectroscopy, X-ray diffraction (XRD), dynamic light scattering (DLS), transmission electron microscopy (TEM) and atomic force microscopy (AFM). The antibacterial efficacy of the synthesized AgNPs was studied against human pathogenic Gram positive (methicillin-resistant Bacillus subtilis, Staphylococcus aureus) and Gram negative (Escherichia coli and Serratia marcescens) bacteria. Isolation and identification of the endophytic fungus Isolation of the endophytic fungus was carried out as described by Hallmann et al., with little modifications. Fresh leaves of R. sativus (Radish) were collected from Sultanpur, Solan district of Himachal Pradesh, India. The collected leaves were thoroughly washed under running tap water for 5 min to remove dust and debris adhering to them. The leaf samples were surface sterilized by immersing sequentially in 70% ethanol for 3 min, 0.5% sodium hypochloride for 1 min and again 70% ethanol for 30 s and finally rinsed three times in sterile distilled water to remove sterilants and then allowed to dry under sterile condition. Then, leaf samples were cut into small pieces and inoculated on potato dextrose agar plates (PDA) and incubated at 28°C for 5 days. Then with a sterile scalpel, the leaves of 0.5 cm size were carefully dissected and placed in Petri plates containing PDA supplemented with streptomycin sulfate (100 mg/l) to suppress the bacterial growth. The plates were sealed using parafilm and incubated at 25 ± 1°C until Figure 1 Microscopic view of endophytic fungus Alternaria sp. endophytic fungi emerged. As and when the hyphal tips emerged out from plant segments they were isolated, subcultured and brought to pure culture by serial subculturing. Lactophenol cotton blue staining method was used for staining the fungal culture and visualized under microscope (Quasmo, India). Identification of fungal endophytes was carried out based on morphotaxonomic characteristics (i.e. the color, shape and growth of cultured colonies), microscopic characteristics (i.e. the structure of hyphae, conidia and conidiophores) and using standard identification manuals. Isolated endophytic fungus ( Fig. 1) is identified as Alternaria sp. Fungal fermentation of Alternaria sp. A loopful of fungus Alternaria sp. was inoculated in two 500 ml Erlenmeyer conical flasks containing 200 ml of fermen-tation medium (Potato dextrose broth), and then incubated at 28°C on a rotary shaker for 72 h. The biomass was harvested after 72 h of growth by filtration through muslin cloth followed by extensive washing with distilled water to remove any medium component. After that, the washed fungal biomass was filtered and weighed. For hot percolation treatment, 10 g of the fungal biomass was boiled with 100 ml of doubledistilled water for 2 h and then kept undisturbed for 10 min. The resultant mixture was then filtered out using Whatman filter paper no. 1 in conical flask. This filtrate solution (namely biomass filtrate) was used for synthesis of AgNPs. Biosynthesis of AgNPs For AgNPs synthesis, 10 ml of mycelium free filtrate obtained from hot percolation treatment was added to 90 ml of 1 mM aqueous silver nitrate (AgNO 3, Sigma-Aldrich) solution in 250 ml Erlenmeyer flask and incubated at 40°C in a hot water bath for 20 min. The color change of AgNO 3 from colorless to dark brown was observed by visual observation. The obtained AgNPs were purified through repeated centrifugation. Characterization of AgNPs Formation of AgNPs was primarily confirmed with UV-visible spectroscopy (UV-1800 Shimadzu, Japan). The absorption maximum of the sample was recorded at a resolution of 1 nm between wavelengths of 300-700 nm. The XRD patterns of the dried AgNPs were analyzed by a Panalytical X-ray (X'Pert 3 Powder, Netherland) diffractometer instrument operating at a voltage of 45 kV and a current of 40 mA with Cu Ka radiation. The AgNPs samples were scanned in the 2h ranges 0-60 in the continuous scan mode and results were compared with the standard powder diffraction card No. 04-0783 of JCPDS (Joint Committee on Powder Diffraction Standards). The crystallite size of the powders was determined from X-ray line broadening using the Scherrer's equation as follows: where D is the average crystallite domain size perpendicular to the reflecting planes, k is a shape factor, k is the X-ray wavelength (1.5418 ), b is the full width at half maximum (FWHM), and h is the diffraction angle. The probable biomolecules involved in the synthesis and stabilization of nanoparticles were recorded by FTIR spectrum using Perki-nElmer spectrometer (Spectrum-One, U.S.A.). DLS measurements for size distribution and zeta potential were carried out using Zetasizer Nano S90 (Malvern Instruments Ltd., U. K.). Morphology, size, selected area electron diffraction (SAED) pattern and elemental analysis were examined using TecnaiG20, Super twin, double tilt, 200 kV TEM instrument (FEI, Netherland) and AFM in non-contact mode. For TEM analysis, samples in the form of thin films were prepared by drop-coating the biosynthesized AgNPs solution on a carbon coated copper grid and drying at room temperature. The interaction of the transmitted electrons resulted in an image formation which is magnified and focused on an imaging device. To investigate the elemental composition of the biosynthesized AgNPs, energy dispersive X-ray (EDX) analysis was carried out using the same instrument. Samples for AFM were prepared by spin-coating the AgNPs solution into the glass slide. The slide was dried at room temperature and subjected to AFM analysis (Nanoscope8, Bruker, Germany). Evaluation of antibacterial activity The synthesized AgNPs were tested for their antibacterial activity against human bacterial pathogens using the standard disk diffusion method. The Gram positive (methicillinresistant Bacillus subtilis, MTCC 441, Staphylococcus aureus, MTCC 740) and Gram negative (Escherichia coli, MTCC 443 and Serratia marcescens, MTCC 97) bacterial pathogens were obtained from Microbial Type Culture Collection (MTCC), Indian Institute of Microbial Technology (IMTECH), Chandigarh. The Freshly cultured bacterial colonies of tested bacteria were used and 100 ml of inoculum was spread on Mueller-Hinton agar plates. A single colony of each test strain was grown overnight in Mueller-Hinton liquid medium on a rotary shaker at 37°C. The inocula were prepared by diluting the overnight cultures with 0.9% NaCl to a 0.5% Mcfarland standard and applied to plates. After that, different concentrations (5, 10, 15, 20 and 25 mg/ml) of AgNPs were loaded on 6 mm size disks. Finally, inhibition zones were measured after 24 h incubation at 37°C. The experiments were performed in triplicate and Student's t-test was used to evaluate statistically significant differences. Confirmation of AgNPs formations UV-visible spectra were recorded to identify the AgNPs synthesis. It was observed that upon addition of the supernatant of Alternaria sp. into AgNO 3, change in color from colorless to dark brown was observed within 20 min, indicating the synthesis of AgNPs (Fig. 2). Nanostructured metallic particles such as silver and gold have free electrons abundance, and they move through conduction and valence band which is responsible for surface plasmon resonance absorption band. The characteristic surface plasmon resonance peak at about 426 nm is shown in Fig. 2, which may correspond to spherical AgNPs. Study of biomolecules capping of AgNPs FTIR spectroscopic measurements were carried out to identify the possible biomolecules responsible for capping and efficient stabilization of AgNPs synthesized by Alternaria sp. extract. The structure of chemical bonding formed on the synthesized AgNPs was examined by FTIR, to elucidate the nature of interaction between endophytic fungal extract and AgNPs. The FTIR spectrum of AgNPs is shown in Fig. 3, which manifests absorption peaks located at the region about 4000 cm 1 and 500 cm 1. FTIR spectrum shows absorption bands at 3145 cm 1, 1597 cm 1, 1402 cm 1, 1109 cm 1 and 1213 cm 1 995 cm 1 and 911 cm 1 699 cm 1 and 504 cm 1 indicating the presence of capping agent with the nanoparticles (Fig. 3). The broad and intense peak at 3145 cm 1 can be attributed to the stretching of AOH group due to inter and intra hydrogen bonding of compounds indicating the presence of phenolic compounds. The band around 1597 cm 1 is associated with vibration of ACC and is assigned to primary and secondary amines and amide I bonds of proteins. The absorption band at around 1402 cm 1 can be attributed to methyls. The band around 1109 cm 1 and 1213 cm 1 can be attributed to CAN stretching vibration of amines indicating the presence of amino acid. The band around 995 cm 1 and 911 cm 1 can be ascribed to CAH stretching vibration indicating the presence of alkene groups. The band at 699 cm 1 and 504 cm 1 can be attributed to alkyl halides stretching. Morphological study of AgNPs TEM has provided insight into the morphology and the details of size of the synthesized nanoparticles (Fig. 5a-c). The synthesized AgNPs were almost spherical in shape with 10-30 nm size range. The bright rings in SAED pattern suggest that the particles are crystalline in nature (Fig. 5d). The EDX profile (Fig. 5e) shows strong silver peaks approximately at 3 keV, which is typical for the absorption of AgNPs due to surface plasmon resonance. The peak at 8 keV corresponds to copper and may be arisen due to the artifact of copper-grid on which the sample was coated. Further the size and morphology of the AgNPs were determined from AFM ( Fig. 6a and b). It can be seen that the AgNPs were monodispersed with average particle size of 4-28 nm which was in close agreement with XRD and TEM results. Stability of AgNPs The DLS measurement as shown in Fig. 7, presented the size distribution (Fig. 7a) of the AgNPs with an average size of $80 nm and stability (Fig. 7b) for the AgNPs at 16.8 mV in zeta potential analysis. The size obtained by the DLS is different because it gives the average size of the particles. Also it could be due to the non-homogeneous dispersion of the samples. The particle size obtained from XRD, TEM, AFM and DLS is different, due to the variation in principles used for measurement. Antibacterial effect of AgNPs The antibacterial activity (as shown in Table 1 and Fig. 8) of the synthesized AgNPs against the tested human pathogens was assessed on the basis of the zone of inhibition. From Fig. 8 and Table 1, it has been observed that the biogenic AgNPs exhibited relatively high antibacterial activity against Gram-positive and Gram-negative bacteria as compared to that of controls (Fungal supernatant and AgNO 3 ). This relatively high antibacterial activity can be attributed to the size and the high surface area of the AgNPs which enabled them to reach easily the nuclear content of bacteria. For the selected bacterial strains, no zone of inhibition was observed for AgNO 3 solution as well as extract. This symbolizes that the antibacterial potential of extract and AgNO 3 at their selected concentrations did not show any zone of inhibition. The irregular release of insufficient concentrations of silver ions from AgNO 3 may be the reason for its restricted efficacy as antimicrobial agents, which can be enhanced using AgNPs because their large surface area makes them highly reactive. It is suggested that AgNPs may attach to the surface of the bacterial cell membrane and release silver ions which may disrupt cell membrane permeability and bacterial DNA replication. In addition, nanoparticles possess extremely large surface area that provides better contact and interaction with bacterial cells. Undoubtedly, the bactericidal activity is due to the silver cations released from AgNPs pertaining to changes in the membrane structure of bacteria, which lead to the increased membrane permeability of the bacteria and finally cell death. It is apparent also from the data that increasing the concentration of the biogenic AgNPs led to a significant increase in the antibacterial activity. Kharwar et al. reported the antibacterial properties of AgNPs produced by endophytic fungi, Aspergillus clavatus which revealed the zone of inhibition of 10 mm in case of E. coli. Kim et al. reported similar effects on E. coli and S. aureus pathogenic strains. AgNPs synthesized by A. flavus were also reported to possess good antibacterial activity against E. coli by Ninganagouda et al.. In spite of the strong antibacterial activity of biogenic AgNPs, further studies are required to investigate their bactericidal effects at a molecular level for potential widening of their applications. Conclusions In this study, an inexpensive, rapid and a single step technique for the synthesis of silver nanoparticles using endophytic fungal supernatant of Alternaria sp. is developed. The results of TEM and AFM have confirmed that the synthesized silver nanoparticles are mostly spherical in shape with an average size of 4-30 nm. The results of XRD, EDS and SAED confirmed the crystalline nature and elemental composition of the synthesized silver nanoparticles. These silver nanoparticles showed antibacterial activity against human pathogenic bacteria. Therefore, fungus mediated nanoparticles could be used as an excellent source against tested bacteria.

A description of the outer automorphism of S_6, and the invariants of six points in projective space We use a simple description of the outer automorphism of S_6 to cleanly describe the invariant theory of six points in P^1, P^2, and P^3. In §1.1-1.2, we give two short descriptions of the outer automorphism of S 6, complete with a proof that they indeed describe an outer automorphism. (Our goal is to have a construction that the reader can fully understand and verify while sitting on a bus, perhaps doodling in the margins.) In §1.3 we give another variation on this theme that is attractive, but more long-winded. The latter two descriptions do not distinguish any of the six points. Of course, these descriptions are equivalent to the traditional one ( §1.4) -there is after all only one nontrivial outer automorphism (modulo inner automorphisms). In §2, we use this to cleanly describe the invariant theory of six points in projective space. This is not just a random application; the descriptions of §1 were discovered by means of this invariant theory. En route we use the outer automorphism to describe five-dimensional representations of S 5 and S 6, §1.5. The outer automorphism was first described by Hlder in 1895. Most verifications use some variation of Sylvester's synthemes, or work directly with generators of S 6 ; nontrivial calculation is often necessary. Other interpretations are in terms of finite geometries, for example involving finite fields with 2, 3, 4, 5, or 9 elements, and are beautiful, but require non-trivial verification. 1. THE OUTER AUTOMORPHISM OF S 6 1.1. First description of the outer automorphism: the mystic pentagons. Consider a complete graph on five vertices numbered 1 through 5. The reader will quickly verify that there are precisely six ways to two-color the edges (up to choices of colors) so that the edges of one color (and hence the other color) form a 5-cycle, see Figure 1. We dub these the six mystic pentagons. Then S 5 acts on the six mystic pentagons by permuting the vertices, giving a map i : S 5 = S {1,...,5} → S {a,...,f } = S 6. This is an inclusion -the kernel must be one of the normal subgroups {e}, A 5, or S 5, but we visually verify that acts nontrivially. Moreover, it is not a usual inclusion as induces permutation (ad)(bc)(ef) -not a transposition. Hence S 6 = S {a,...,f } acts on the six cosets of i(S 5 ), inducing a map f : S {a,...,f } → S {1,...,6}. This is the outer automorphism. This can be verified in several ways (e.g., (ad)(bc)(ef) induces the nontrivial permutation ∈ Date: Hallowe'en, 2007. 1991 Partially supported by NSF CAREER/PECASE Grant DMS-0228011. S {1,...,6}, so f is injective and hence an isomorphism; and i is not a usual inclusion, so f is not inner), but for the sake of simplicity we do so by way of a second description. There are six ways of dividing the triangles into two sets of 10 so that (i) any two disjoint triangles have opposite colors, and (ii) every tetrahedron has two triangles of each color. (The bijection between these and the mystic pentagons a,..., f is as follows. The triangle 6AB is colored the same as edge AB. The triangle CDE (6 = C, D, E) is colored the opposite of the "complementary" edge AB, where {A, B} = {1,..., 5} − {C, D, E}.) The S 6 -action on this set is the outer automorphism of S 6. (Reason: induces a nontrivial permutation (ad)(bc)(ef) of the mystic pentagons, so the induced map S 6 → S {a,...,f } ∼ = S 6 is injective and hence an isomorphism. But does not induce a transposition on {a,..., f}, so the automorphism is not inner.) This isomorphism S {1,...,6} → S {a,...,f } is inverse to the isomorphism f of §1.1. Another description: labeled icosahedra. Here is another description, which is pleasantly S 6 -symmetric. Up to rotations and reflections, there are twelve ways to number the vertices of an icosahedron 1 through 6, such that antipodal vertices have the same label. Each icosahedron gives ten triples in {1,..., 6}, corresponding to the vertices around its faces. These twelve icosahedra come in six pairs, where two icosahedra are "opposite" if they have no triples in common. (It is entertaining to note that if an icosahedron is embedded in Q() 3 with vertices at (±1, ±, 0), (0, ±1, ±), and (±, 0, ±1), then conjugation in Gal(Q()/Q) sends the icosahedron to its opposite. Here is the golden section.) Then S 6 acts on these six pairs, and this is the outer automorphism. One may show this via bijections to the descriptions of §1.1 and §1.2. Each pair of mystic icosahedra corresponds to two-coloring the triangles in {1,..., 6}, as in §1.2. For the bijection to §1.1, the cyclic order of the vertices around vertex 6 gives a mystic pentagon. (This provides a hands-on way of understanding the S 6 -action on the mystic pentagons.) This description is related to the explanation of the outer automorphism by John Baez in . 1.4. Relation to the usual description of the outer automorphism of S 6. The usual description of the outer automorphism is as follows (e.g. ). A syntheme is a matching of the numbers 1,..., 6, i.e. an unordered partition of {1,..., 6} into three sets of size two. A pentad is a set of five synthemes whose whose union is the set of all 15 pairs. Then there are precisely six pentads, and the action of S 6 on this set is via the outer automorphism of S 6. We explain how to get pentads from the mystic pentagons. Each mystic pentagon determines a bijection between the white edges and the black edges, where edge AB corresponds with edge CD if AB and CD don't share a vertex. If E = {1,..., 5} − {A, B, C, D}, then to each such pair we obtain the syntheme AB/CD/E6, and there are clearly five such synthemes, no two of which share an edge, which hence form a pentad. For example, mystic pentagon a yields the pentad Another common description of the outer automorphism relates directly to Figure 1. We find a subgroup G < S 5 of size 20; we take the subgroup preserving figure a of Figure 1. Then S 5 acts transitively on the six cosets of G, giving a map i : S 5 → S 6. This map is an inclusion as is not in its kernel. Then S 6 acts (transitively) on the six cosets of i(S 5 ), yielding a map : S 6 → S 6. The image (as it is transitive) has size > 2, hence (as S 6 has only 3 normal subgroups) the kernel is e, hence is an automorphism. Then it is not inner, as i(S 5 ) is not one of the six "obvious" S 5 's in S 6. 1.5. Representations of S 5 and S 6. In Figure 1, the edges are colored black and white so that each edge appears in each color precisely three times with this choice. This has the advantage that any odd permutation in S 5 (or S 6 ) permutes the six pentagons and exchanges the colors. The pentagons give a convenient way of understanding the two 5-dimensional irreducible representations of S 5. The permutation representation induced by this S 5 action on the mystic pentagons splits into an irreducible 5-dimensional representation F 5 and a trivial representation 1. The other irreducible 5-dimensional S 5 -representation F 5 is obtained by tensoring F 5 with the sign representation, which can be interpreted as the S 5 action on the mystic pentagons "with sign corresponding to color-swapping". There are four irreducible 5-dimensional representation of S 6. One is the standard representation (which we here denote B 5 ), obtained by subtracting the trivial representation 1 from the usual permutation representation. A second is obtained by tensoring with the sign representation : B 5 := B 5 ⊗. A third is analogous to the standard representation, obtained by subtracting the trivial representation 1 from the (outer) permutation representation of S 6 on the six mystic pentagons. One might denote this the outer automorphism representation. The fourth 5-dimensional S 6 -representation is O 5 := O 5 ⊗. One might term this the signed outer automorphism representation. It is clear from the construction that F 5 and F 5 are obtained by restriction from O 5 and O 5. 1.6. An alternate description of these representations is as follows. There are twelve 5cycles on vertices labeled {1,..., 5}, which come in pairs of "opposites", consisting of disjoint 5-cycles. Each mystic pentagon is equivalent to such a pair. If x is a 5-cycle, denote its opposite by x. (The same construction applies for triangles on {1,..., 6}, §1.2, or labeled icosahedra, §1.3.) If we have twelve variables Z a,..., Z f, Z a,..., Z f, with the conditions Z x = −Z x, the S 6 -action induces the representation O 5 ⊕. (Of course, the other three representations can be described similarly, O 5 by imposing Z x = Z x instead and replacing by 1, F 5 by considering the S 5 -action, and F 5 by making both changes.) THE INVARIANT THEORY OF SIX POINTS IN PROJECTIVE SPACE We will now relate the outer automorphism of S 6 to the space of six ordered points in projective space, or more precisely the geometric invariant theory quotient (P n ) 6 //P GL(n+ 1). The algebraic statements in this section may be readily checked by any computer algebra program such as Maple, so the details are omitted. They were derived using explicit representation theory of S 6, and again the details are unenlightening and will be omitted. In some sense these quotients generalize the notion of cross-ratio, the space of four points in P 1. Any two generic sets of six ordered points in P n are projectively equivalent if n > 3, so the interesting cases are n = 1, 2, 3. All three cases were studied classically, and were known to behave beautifully. 2.1. Six points on P 1. The space of six points on P 1 may be interpreted as a threefold in P 5 cut out by the equations Z 1 + + Z 6 = Z 3 1 + + Z 3 6 = 0. (Aside: This is one of the many ways in which 6 points are special. If m = 6, the space of m points in P 1, (P 1 ) m //P GL, is cut out by quadrics .) This is the Segre cubic relation, and this moduli space is known as the Segre cubic threefold, which we denote S 3. There is an obvious S 6 -action on both (P 1 ) 6 and the variables Z 1,..., Z 6. One might hope that these actions are conjugate, which would imply some bijection between the six points and the six variables. But remarkably, they are related by the outer automorphism of S 6. An alternate interpretation of this quotient is as the space of equilateral hexagons in real 3-space, with edges labeled 1 through 6 cyclically, up to translations and rotations . Rearranging the order of the edges induces a permutation of the Z-variables via the outer automorphism. Here is a clue that the outer automorphism is relevant. The cross-ratio of a certain four of the six points is given by → −(Z 1 +Z 2 )/(Z 3 +Z 4 ). A more symmetric avatar of the cross-ratio of four points on a line is given by (Here points of P 1 are written in projective coordinates for convenience; more correctly we should write 1].) Note that the S 4 -symmetry is clear in this manifestation. The image is the line X + Y + Z = 0 in P 2. The traditional cross-ratio is −X/Y. In this symmetric manifestation, the cross-ratio of a certain four of the six points is given by → . The correspondence of a pair of points with a "syntheme" of the Z-variables is a hint that the outer automorphism is somehow present. We now describe the moduli map (P 1 ) S 3 explicitly. If the points are p 1,..., p 6 (1 ≤ i ≤ 6), the moduli map is given (in terms of the second description of the outer automorphism, §1.2) by.., f}, and the sign is +1 if triangle ABC is black, and −1 if the triangle is white. Note that Z x = 0. As an added bonus, we see that the S 6 -representation on H 0 (S 3, O) is the signed outer automorphism representation O 5 (see §1.6). Six points in P 3, and the Igusa quartic. The Geometric Invariant Theory quotient of six points on P 3 is the Igusa quartic threefold I 4. To my knowledge, the presence of the outer automorphism was realized surprisingly recently, by van der Geer in 1982 (in terms of the two isomorphisms of Sp(4, F 2 ) with S 6, , see also ): w a + + w f = 0, (w 2 a + + w 2 f ) 2 − 4(w 4 a + + w 4 f ) = 0. (Igusa's original equation obscured the S 6 -action.) Via the Gale transform (also known as the "association map"), this is birational to the space of six points on P 1, where six distinct points in P 1 induce six points on P 3 by placing them on a rational normal curve (e.g. via p i → ), and six general points on P 3 induce six points on P 1 by finding the unique rational normal curve passing through them. We describe the rational map (P 1 ) 6 I 4, and then the rational map (P 3 ) 6 I 4. The rational map (P 1 ) 6 I 4 is described as follows, using the first description of the outer automorphism, §1.1: where N = 2 if the edge BC has the same color as edge DE, and N = −1 otherwise. (A quick inspection of the mystic pentagons shows that W x = 0.) Hence the S 6representation on H 0 (I 4, O) is O 5, the outer automorphism representation. In terms of the usual description of the outer automorphism §1.4, N = −1 if 6A/BC/DE is a syntheme in the pentad, and 2 otherwise. The birationality to the Segre cubic S 3 arises by projective duality (S 3 and I 4 are dual hypersurfaces), which should not involve the outer automorphism. Indeed, the duality map S 3 I 4 is given by Z 2 y, and the duality map I 4 S 3 is given by It is perhaps surprising that these moduli maps are somehow "dual", while the corresponding S 6 -representations H 0 (S 3, O) ∼ = O 5 and H 0 (I 4, O) ∼ = O 5 are not dual. However, their projectivizations are dual, as they differ by a sign representation. Here S 5 acts by the "outer action" corresponding to x on the six points {1,..., 6}, and S 4 acts by permuting the co-ordinates {w, x, y, z} and by sign. (There is significant abuse of notation in the way the formula is presented, but hopefully the meaning is clear.) This formula is less horrible than it appears, as the summands can be interpreted as attractive geometric configurations on the icosahedra of §1.3. 2. 3. Six points in P 2. Finally, we describe the invariants of six points in P 2 in terms of the mystic pentagons. This quotient is a double cover of P 4 branched over the Igusa quartic I 4. The Gale transform sends six points in P 2 to six points in P 2, and exchanges the sheets. The branch locus of this double cover (the "self-associated" sextuples in the language of the Gale transform) corresponds to when the six points lie on a conic; by choosing an isomorphism of this conic with P 1, the rational map (P 1 ) 6 I 4 is precisely the moduli map described above in §2.2. Suppose the points in P 2 are (1 ≤ i ≤ 6). We describe the moduli map (P 2 ) 6 P 4 in terms of the mystic pentagons §1.1: Corresponding to each term are two edges (corresponding to the pairs AB, CD, EF not containing 6). Then N = 2 if the two edges have the same color, and −1 otherwise. Notice the similarity to the moduli map for the Igusa quartic above, in §2.2; this is not a coincidence, and we have chosen the variable names W x for this reason. Again, N = −1 if 6A/BC/DE is a syntheme in the pentad, and 2 otherwise. The condition for six points to be on a conic is for their image on the Veronese embedding to be coplanar, hence that the following expression is 0: x 2 6 y 2 6 z 2 6 x 6 y 6 y 6 z 6 z 6 x 6  . (Alternatively, the vanishing of this determinant ensures the existence of a nontrivial quadric x 2 x 2 + y 2 y 2 + z 2 z 2 + xy xy + yz yz + zx zx = 0 satisfied by (x i, y i, z i ) for all i between 1 and 6.) Then the formula for the double fourfold that is the Geometric Invariant Theory quotient of six points on P 2 is W 2 x 2 − 4 W 4 x + 324V 2 = 0 and it is clear that it is branched over the Igusa quartic I 4. (See for more information.) 2.4. Relation to the usual description of the invariants of six points on P 1. We relate the explicit invariant theory of §2.1 to the classical or usual description of the invariants of six points on P 1. In the matching diagram language of (basically that of Kempe in 1894), the ring of invariants is generated by the variables where {A,..., F } = {1,..., 6}. The variables Z x of the Segre cubic threefold S 3 are related to the matching diagrams in a straightforward way: X 13 26 45 = (Z a + Z b )/2 (and similarly after application of the S 6 -action on both sides). Notice that under the outer automorphism, pairs are exchanged with "synthemes" (= partitions into three pairs), and that is precisely what we see here. This can of course be easily inverted, using Z a = (Z a + Z b )/2 + (Z a + Z c )/2 − (Z b + Z c )/2. As the X-variables form a 15-dimensional vector space with many relations, there are many formulas for the Z-variables in terms of the X-variables.

Mild forms of herpes encephalitis. Three cases of herpes encephalitis are described. A definite diagnosis was established and all patients made a good recovery without specific antiviral chemotherapy. These reports are representative of those forms of herpes encephalitis with a good prognosis. It is suggested that the introduction of rapid non-invasive procedures will indicate a higher incidence of herpes encephalitis than is presently accepted. The relevance of repeated episodes of herpes encephalitis with respect to the aetiology of some psychiatric disorders and in the evaluation of antiviral agents is also discussed.

In the Shape of a Goddess I walked into Hughs house. I knock this time, lightly on the front door, and then walk in. His home looks like an abandoned art studio or woodworking shop, and its hot inside, overheated; the thermostat must be broken. There is hardly room to maneuver through the machinery and lumber that fills the space. The past years mail is piled up on the band saw, and several months of dirty dishes sit on the table in the corner. Sawdust covers the hardwood floors, mixed with pistachio nut shells, cat shit, and broken bits of pottery. Beneath the floor, I can hear Ramses in the basement, barking and whining to be set free, while two or three generations of cats surround my feet and meow to be fed. Two years ago, Hugh had decided to leave this house and move back to his farm. He took me there many times. A lonely placeacres of beautiful flowering fields where once horses grazed now filled with the hum of insects and the soft erratic flight of butterflies. Inside the old ramshackle farmhouse, dust had settled on the kitchen countertops, the near-empty bookshelves, the plywood floor. A cold, black wood-burning stove stood dormant in the middle of the one large open room, its chimney reaching up through the roof where the insulation hung down in big loops.The childrens rooms were up in the loft. Dolls and books and toys were left behind. We would take Ramses to the farm with us. It was his chance to run like mad through the fields. Hugh had wanted to go back. Hed had enough of this backward college town where nobody understood him, so he gathered all the beautiful antique furniture inherited from his aunt, packed up a U-haul, and moved it to the farm. I just want to be alone, he said. He took everything except his brass bed and his four-cup coffee maker. But after moving the furniture out, he never bothered to leave himself. Instead, he moved his sculptures in. They told him at the art studio on

(CNN) -- James Karl Buck helped free himself from an Egyptian jail with a one-word blog post from his cell phone. James Karl Buck sent a message using Twitter which helped get him out of an Egyptian jail. Buck, a graduate student from the University of California-Berkeley, was in Mahalla, Egypt, covering an anti-government protest when he and his translator, Mohammed Maree, were arrested April 10. On his way to the police station, Buck took out his cell phone and sent a message to his friends and contacts using the micro-blogging site Twitter. The message only had one word. "Arrested." Within seconds, colleagues in the United States and his blogger-friends in Egypt -- the same ones who had taught him the tool only a week earlier -- were alerted that he was being held. Twitter is a social-networking blog site that allows users to send status updates, or "tweets," from cell phones, instant messaging services and Facebook in less than 140 characters. Hossam el-Hamalawy, a Cairo-based blogger at UC-Berkeley, was one of the people who got word of Buck's arrest. "At first I was worried about his safety," el-Hamalawy said. But Buck was able to send updates every couple of hours saying he was still detained, he had spoken to the prosecutor, he still had not been charged, and he was worried about Maree. "Usually the first thing the police go for is the detainees' cameras and cellular phones," el-Hamalawy said. "I'm surprised they left James with his phone." But Biz Stone, co-founder of Twitter, said he and others knew that the service could have wide-reaching effects early on, when the San-Francisco, California-based company used it to communicate during earthquakes. Stone said that as the service got more popular, they began to hear stories of people using Twitter during natural disasters with a focus on activism and journalism. Buck's urgent message is proof of the value of Twitter, Stone said. Buck's entry set off a chain of events that led to his college hiring a lawyer on his behalf. "James' case is particularly compelling to us because of the simplicity of his message -- one word, 'arrested' -- and the speed with which the whole scene played out," Stone said. "It highlights the simplicity and value of a real-time communication network that follows you wherever you go." Initially, the Twitter message was a precaution -- something people could trace in case anything went wrong, Buck said. "The most important thing on my mind was to let someone know where we were so that there would be some record of it ... so we couldn't [disappear]," Buck said. "As long as someone knew where we were, I felt like they couldn't do their worst [to us] because someone, at some point, would be checking in on them." Buck began using Twitter as a way to keep up in touch with the bloggers at the heart of his project and the events going on in Egypt that he intended to cover. Buck was working on a multimedia project on Egypt's "new leftists and the blogosphere" as part of his master's degree thesis. Buck found out from a Twitter message that a planned protest against rising food prices and decreasing wages in Mahalla had been shut down by Egyptian authorities April 6. The next day, tensions rose as family and friends of protesters who had been detained took to the streets, eventually throwing Molotov cocktails and setting tires on fire, he said. On April 10, Buck returned to Mahalla, where protests continued. "I was worried about getting arrested, so I made sure to stay at a distance from the protest so there was no way I could be accused of being part of it," Buck said. "Mohammed and I had a bad sense; it was really tense." When the men tried to escape, they were detained. That's when Buck thought of Twitter and sent out his message. Buck and Maree were interrogated, released and then detained again by the same police officers. "We are really worried that we are off the radar now," Buck said. Eventually Buck was released, but Maree was transferred to another police station. As he left the station, Buck reached into his pocket, as he did less than 24 hours earlier. Another one-word blog entry said it all: "Free." As happy as he was to be free, Buck said, his biggest frustration was leaving behind the translator who helped protect him during the riots. Although the Twitter message helped him find contacts to get out of prison, he says it was more the power of the network he had as an American that enabled him to be released so quickly. "Mohammed was sitting next to me," he said. "But he didn't have the network to call. I tried to use my network to shield him until they tore us apart." Twitter may not have been able to secure Maree's release, but Buck hopes his initial reason for using Twitter will help find his missing friend. "It was my big hope that people would get [the message] right away and at least put a thumbtack on the map as far as our location," Buck said. There has been no official confirmation regarding Maree's whereabouts. Attiya Shakran, press counsel for the Egyptian Consulate in San Francisco, said Maree was released April 13. Maree's brother Ahmed Maree said that he had not heard from his brother and that he believes he is still in jail. Government officials in Egypt could neither confirm nor deny Maree's release, despite repeated requests for comment. Buck is now using his story and Twitter page as a way to rally people looking for answers about Maree's status. He's gone as far as publishing the phone number of the press counsel of the Egyptian Consulate in San Francisco and posting a petition for Maree's release. CNN Cairo's Housam Ahmed and Aneesh Raman contributed to this report.

Two candidates are vying in the Democratic primary for the Ward 8 seat on the Akron City Council. They are Shammas Malik, an attorney who recently stepped down from his position with the city law department to run for office, and the Rev. Curtis T. Walker Sr. of the A.M.E. Zion Church, the president of the Akron Board of Education. Each would add to the quality of the council. We recommend the election of Shammas Malik on May 7. Malik is just three years out of law school, yet he already has begun to make a mark, returning from Harvard to the city where he grew up. At the law department, he worked in the civil division, handling a variety of matters, getting to know how the city works and building relationships. He is a quick study, and it shows in the depth with which he talks about the city and its challenges. In this campaign, Malik has made a priority of the Merriman Valley, with an eye on enhancing the area’s place as a gateway to the Cuyahoga Valley National Park. He emphasizes public safety, especially improved training for police officers and building better relationships between the department and neighborhoods. He grasps the complexity of economic development, the need for a strong downtown and greater opportunity for minorities. Malik knows the city must attract families, and in that way, his relative youth is representative of the need to refresh the city population and its leadership. He would be a strong addition to the City Council. So would the Rev. Walker. The past two decades, he has been an effective and positive presence on the school board. His calm and thoughtful approach would be helpful on a council afflicted with mistrust and, at times, dysfunction. He would be a voice for economic inclusion, the black community long on the outside. He would aid in securing the key relationship between the city and the schools. He cites the importance of greater diversity on the police force, recreation programs for young people and a vibrant downtown. Walker shares many of the priorities of his opponent. The edge goes to Shammas Malik for his new voice and time spent in city government. Two candidates are vying in the Republican primary for the Ward 8 seat on the Akron City Council. They are Greg Dunham, a retired accountant from Ohio Edison, and Brian Fortney, global project manager for Rockwell Automation. As in the Democratic primary, this race features two solid candidates. We recommend the election of Brian Fortney on May 7. Fortney frames the priorities for Ward 8 around protecting home values. Thus, he emphasizes public safety, good schools and strong city services. Just as important as his focus on the Northwest Akron ward is his grasp of the larger challenges of the city, including its strained finances. His business experience would be a plus as the city looks to improve its operations and implement its economic development plans. His candidacy also benefits from his community involvement, including time on the boards of the Akron Symphony, Torchbearers and Actors’ Summit. Such relationships add to his understanding of the city. Dunham, who ran unsuccessfully for the U.S. House in 2018, brings his own strengths. As an accountant, he would be helpful as the council examines city finances. He sees a need for improved transparency and accountability in city operations, along with working toward a larger reserve fund. He long has been active in the ward, currently serving as the treasurer of the Fairlawn Heights Neighborhood Association, for which a leading priority is slowing speeders on residential streets.

After picking up a glass of wine and a name tag at the bar, everyone was assigned to a curved booth. At the sound of the bell, each person had two minutes to introduce themselves to a stranger. The lights were dimmed and there were candles, but it wasn't speed-dating. In a social networking age when it's effortless to claim hundreds of friends, the 30 women gathered after work at Roe lounge in San Francisco were looking to start authentic friendships by meeting face to face. Ordained pastor and life coach Shasta Nelson, 33, launched GirlfriendCircles.com this summer, after realizing that it's possible for women to find jobs, dates or apartments online but that there wasn't a way to find simple friendship. "I moved to San Francisco a year ago, and I'd see these small groups of women friends in cafes laughing together, and I was jealous, I wanted to ask if I could be their friend, but there really isn't a way for adult women to approach one another like that," she said. For a $30, six-month membership fee, women join GirlfriendCircles.com and are invited to cafes to meet small groups of "connecting circles" of women who match their age, neighborhood and schedule. Afterward they can establish friendships with the women they meet or post their own events on the site, such as home wine tastings, cooking classes or movie nights. There are 550 members in the Bay Area, ranging in age from 21 to 67. In September, Nelson's Web site expanded to Seattle, Chicago and Portland. Los Angeles and New York were added this month. Members are career women who just moved to a new city, women who lost a job and with it their only social circle, divorcees who lost their friends in the breakup, women who work from home or in a male-dominated field or women whose close friends married and fell out of touch. "I wanted to meet more moms," said Monica Fitzsimmons, 39, who attended the speed-friending event. "I met someone I liked, and just last weekend we got together with our kids," she said. Ciana Wilson of Berkeley, 26, tried speed-friending after tiring of "textual friendships." She estimated she spends 90 percent of her friend time either texting, IM-ing, reading about buddies on Facebook or e-mailing - but not actually with them. At Roe, Wilson shared a two-minute conversation with a woman who had lived in New York and spent a little time in Paris. Wilson, who recently earned her master's degree from New York University in French studies, brightened and made notes on her speed-friending form. "It can seem a bit awkward to meet friends this way," she said. "But I feel like technology is changing people's lives so much that the nature of making friends has to change, too," she said. At the end of the event, women jotted down the names of women they would like to meet a second time. Nelson collected the forms, and if both women chose each other, they will be given each others' e-mail addresses. Although her Web site is only a few months old, Nelson has been able to conduct a few exit surveys when members close their accounts. The majority say it's because they made enough friends. Nelson, who worked as a pastor in Seventh-day Adventist churches in Michigan after graduating in 2000 from divinity school, says she has now found her calling. Her right-hand man is her father, Jeff Emery, a computer engineer in Michigan. His job was downsized to four days a week, giving him time to build her Web site. GirlfriendCircles.com allows her to promote community, she said. Nelson previously started a spirituality group called Second Wind. "Besides genetics, exercise and friendships are the two things that determine longevity," she said. "We place so much importance on exercise - can you imagine what kind of world this would be if we paid that much attention to our circle of friends?" GirlfriendCircles.com hosts another speed-friending event 7-9 p.m. Dec. 8 at A Perfect Finish Wine Bar, 55 S. First St., San Jose. $12-$20; visit speedfriendingsb.eventbrite.com to order tickets.

Loss of Crown Immunity in the NHS; the effect on hotel services. A less than "holy" combination of the Environmental Officers of Health (EHO) and the pest control industry had for some time been conducting a campaign for the removal of Crown Immunity from hospitals. They had not been so vociferous in their campaign to remove the same privilege from prisons and the armed services. I find it difficult to believe this was out of conviction that other institutions were trouble free. For in 1985 of the 191 Institutional outbreaks only 37 (19%) were in hospitals. The outbreak of Salmonella food poisoning at the Stanley Royd hospital provided enough public pressure to be generated for crown immunity to be removed from NHS premises. It remained in prisons and the armed services.

Monte Carlo simulation of minority carrier transport and light emission phenomena in GaAs devices We present a theoretical study of hot-carrier-induced light emission in III-V semiconductor devices. Carrier heating in the intense electric fields present under high-bias conditions is studied using self-consistent Monte Carlo simulations. The carrier distribution functions obtained from the simulation are then incorporated into a pseudopotential algorithm that describes the direct optical transitions and calculates the corresponding spectra. We show that, in general, no straightforward extrapolation of 'carrier temperatures' from the slope of the measured spectra is possible. Our analysis indicates that light emission due to hot carriers is made up by direct radiative interband transitions within the conduction and valence bands as well as between them.

Phenomenon in the US of making accusations of subversion or treason without evidence U.S. anti-communist propaganda of the 1950s, specifically addressing the entertainment industry McCarthyism is the practice of making accusations of subversion or treason without proper regard for evidence.[1] The term refers to U.S. senator Joseph McCarthy (R-Wisconsin) and has its origins in the period in the United States known as the Second Red Scare, lasting from the late 1940s through the 1950s.[2] It was characterized by heightened political repression and a campaign spreading fear of Communist influence on American institutions and of espionage by Soviet agents.[2] What would become known as the McCarthy era began before McCarthy's term in 1953. Following the First Red Scare, in 1947, President Truman signed an executive order to screen federal employees for association with organizations deemed "Totalitarian, Fascist, Communist, or subversive", or advocating "to alter the form of Government of the United States by unconstitutional means." In 1949, a high-level State Department official was convicted of perjury in a case of espionage, and the Soviet Union tested an atomic bomb. The Korean War started the next year, raising tensions in the United States. In a speech in February 1950, Senator McCarthy presented an alleged list of members of the Communist Party working in the State Department, which attracted press attention. The term "McCarthyism" was published for the first time in late March of that year in the Christian Science Monitor, and in a political cartoon by Herblock in the Washington Post. The term has since taken on a broader meaning, describing the excesses of similar efforts. In the early 21st century, the term is used more generally to describe reckless, unsubstantiated accusations, and demagogic attacks on the character or patriotism of political adversaries. During the McCarthy era, hundreds of Americans were accused of being Communists or Communist sympathizers; they became the subject of aggressive investigations and questioning before government or private industry panels, committees, and agencies. The primary targets of such suspicions were government employees, those in the entertainment industry, academicians, and labor-union activists. Suspicions were often given credence despite inconclusive or questionable evidence, and the level of threat posed by a person's real or supposed leftist associations or beliefs were sometimes exaggerated. Many people suffered loss of employment or destruction of their careers; some were imprisoned. Most of these punishments came about through trial verdicts that were later overturned,[3] laws that were later declared unconstitutional,[4] dismissals for reasons later declared illegal[5] or actionable,[6] or extra-legal procedures, such as informal blacklists, that would come into general disrepute. The most notable examples of McCarthyism include the so-called investigations conducted by Senator McCarthy, and the hearings conducted by the House Un-American Activities Committee (HUAC). Origins [ edit ] McCarthyism was in a cartoon by Washington Post, March 29, 1950. One of the earliest uses of the termwas in a cartoon by Herbert Block ("Herblock"), published in the, March 29, 1950. President Harry S. Truman's Executive Order 9835 of March 21, 1947, required that all federal civil-service employees be screened for "loyalty". The order said that one basis for determining disloyalty would be a finding of "membership in, affiliation with or sympathetic association" with any organization determined by the attorney general to be "totalitarian, Fascist, Communist or subversive" or advocating or approving the forceful denial of constitutional rights to other persons or seeking "to alter the form of Government of the United States by unconstitutional means."[7] The historical period that came to be known as the McCarthy era began well before Joseph McCarthy's own involvement in it. Many factors contributed to McCarthyism, some of them with roots in the First Red Scare (1917–20), inspired by Communism's emergence as a recognized political force and widespread social disruption in the United States related to unionizing and anarchist activities. Owing in part to its success in organizing labor unions and its early opposition to fascism, and offering an alternative to the ills of capitalism during the Great Depression, the Communist Party of the United States increased its membership through the 1930s, reaching a peak of about 75,000 members in 1940–41.[8] While the United States was engaged in World War II and allied with the Soviet Union, the issue of anti-communism was largely muted. With the end of World War II, the Cold War began almost immediately, as the Soviet Union installed Communist puppet régimes in areas it had occupied across Central and Eastern Europe. The United States backed anti-communist forces in Greece and China. Although the Igor Gouzenko and Elizabeth Bentley affairs had raised the issue of Soviet espionage in 1945, events in 1949 and 1950 sharply increased the sense of threat in the United States related to Communism. The Soviet Union tested an atomic bomb in 1949, earlier than many analysts had expected, raising the stakes in the Cold War. That same year, Mao Zedong's Communist army gained control of mainland China despite heavy American financial support of the opposing Kuomintang. Many U.S. policy people did not fully understand the situation in China, despite the efforts of China experts to explain conditions. In 1950, the Korean War began, pitting U.S., U.N., and South Korean forces against Communists from North Korea and China. During the following year, evidence of increased sophistication in Soviet Cold War espionage activities was found in the West. In January 1950, Alger Hiss, a high-level State Department official, was convicted of perjury. Hiss was in effect found guilty of espionage; the statute of limitations had run out for that crime, but he was convicted of having perjured himself when he denied that charge in earlier testimony before the HUAC. In Britain, Klaus Fuchs confessed to committing espionage on behalf of the Soviet Union while working on the Manhattan Project at Los Alamos National Laboratory during the War. Julius and Ethel Rosenberg were arrested in 1950 in the United States on charges of stealing atomic-bomb secrets for the Soviets, and were executed in 1953. Other forces encouraged the rise of McCarthyism. The more conservative politicians in the United States had historically referred to progressive reforms, such as child labor laws and women's suffrage, as "Communist" or "Red plots", trying to raise fears against such changes.[9] They used similar terms during the 1930s and the Great Depression when opposing the New Deal policies of President Franklin D. Roosevelt. Many conservatives equated the New Deal with socialism or Communism, and thought the policies were evidence of too much influence by so-called Communist policy makers in the Roosevelt administration.[10] In general, the vaguely defined danger of "Communist influence" was a more common theme in the rhetoric of anti-communist politicians than was espionage or any other specific activity. Senator Joseph McCarthy McCarthy's involvement in these issues began publicly with a speech he made on Lincoln Day, February 9, 1950, to the Republican Women's Club of Wheeling, West Virginia. He brandished a piece of paper, which he claimed contained a list of known Communists working for the State Department. McCarthy is usually quoted as saying: "I have here in my hand a list of 205—a list of names that were made known to the Secretary of State as being members of the Communist Party and who nevertheless are still working and shaping policy in the State Department."[11] This speech resulted in a flood of press attention to McCarthy and helped establish his path to becoming one of the most recognized politicians in the United States. The first recorded uses of the term "McCarthyism" were in the Christian Science Monitor on March 28, 1950 ("Their little spree with McCarthyism is no aid to consultation");[12] and then, on the following day, in a political cartoon by Washington Post editorial cartoonist Herbert Block (Herblock). The cartoon depicts four leading Republicans trying to push an elephant (the traditional symbol of the Republican Party) to stand on a platform atop a teetering stack of ten tar buckets, the topmost of which is labeled "McCarthyism". Block later wrote: "nothing [was] particularly ingenious about the term, which is simply used to represent a national affliction that can hardly be described in any other way. If anyone has a prior claim on it, he's welcome to the word and to the junior senator from Wisconsin along with it. I will also throw in a set of free dishes and a case of soap."[13] Institutions [ edit ] A number of anti-communist committees, panels, and "loyalty review boards" in federal, state, and local governments, as well as many private agencies, carried out investigations for small and large companies concerned about possible Communists in their work forces. In Congress, the primary bodies that investigated Communist activities were the HUAC, the Senate Internal Security Subcommittee, and the Senate Permanent Subcommittee on Investigations. Between 1949 and 1954, a total of 109 investigations was carried out by these and other committees of Congress.[14] On December 2, 1954, the United States Senate voted 65 to 22 to condemn McCarthy for "conduct that tends to bring the Senate into dishonor and disrepute". Executive branch [ edit ] Loyalty-security reviews [ edit ] Executive Order 9835, signed by President Truman in 1947 In the federal government, President Truman's Executive Order 9835 initiated a program of loyalty reviews for federal employees in 1947. It called for dismissal if there were "reasonable grounds ... for belief that the person involved is disloyal to the Government of the United States."[15] Truman, a Democrat, was probably reacting in part to the Republican sweep in the 1946 Congressional election and felt a need to counter growing criticism from conservatives and anti-communists.[16] When President Dwight Eisenhower took office in 1953, he strengthened and extended Truman's loyalty review program, while decreasing the avenues of appeal available to dismissed employees. Hiram Bingham, chairman of the Civil Service Commission Loyalty Review Board, referred to the new rules he was obliged to enforce as "just not the American way of doing things."[17] The following year, J. Robert Oppenheimer, scientific director of the Manhattan Project that built the first atomic bomb, then working as a consultant to the Atomic Energy Commission, was stripped of his security clearance after a four-week hearing. Oppenheimer had received a top-secret clearance in 1947, but was denied clearance in the harsher climate of 1954. Similar loyalty reviews were established in many state and local government offices and some private industries across the nation. In 1958, an estimated one of every five employees in the United States was required to pass some sort of loyalty review.[18] Once a person lost a job due to an unfavorable loyalty review, finding other employment could be very difficult. "A man is ruined everywhere and forever," in the words of the chairman of President Truman's Loyalty Review Board. "No responsible employer would be likely to take a chance in giving him a job."[19] The Department of Justice started keeping a list of organizations that it deemed subversive beginning in 1942. This list was first made public in 1948, when it included 78 groups. At its longest, it comprised 154 organizations, 110 of them identified as Communist. In the context of a loyalty review, membership in a listed organization was meant to raise a question, but not to be considered proof of disloyalty. One of the most common causes of suspicion was membership in the Washington Bookshop Association, a left-leaning organization that offered lectures on literature, classical music concerts, and discounts on books.[20] J. Edgar Hoover and the FBI [ edit ] FBI director J. Edgar Hoover designed President Truman's loyalty-security program, and its background investigations of employees were carried out by FBI agents. This was a major assignment that led to the number of agents in the bureau being increased from 3,559 in 1946 to 7,029 in 1952. Hoover's sense of the Communist threat and the standards of evidence applied by his bureau resulted in thousands of government workers losing their jobs. Due to Hoover's insistence upon keeping the identity of his informers secret, most subjects of loyalty-security reviews were not allowed to cross-examine or know the identities of those who accused them. In many cases, they were not even told of what they were accused.[21] Hoover's influence extended beyond federal government employees and beyond the loyalty-security programs. The records of loyalty review hearings and investigations were supposed to be confidential, but Hoover routinely gave evidence from them to congressional committees such as HUAC.[22] From 1951 to 1955, the FBI operated a secret "Responsibilities Program" that distributed anonymous documents with evidence from FBI files of Communist affiliations on the part of teachers, lawyers, and others. Many people accused in these "blind memoranda" were fired without any further process.[23] The FBI engaged in a number of illegal practices in its pursuit of information on Communists, including burglaries, opening mail, and illegal wiretaps.[24] The members of the left-wing National Lawyers Guild were among the few attorneys who were willing to defend clients in communist-related cases, and this made the NLG a particular target of Hoover's. The office of this organization was burgled by the FBI at least 14 times between 1947 and 1951.[25] Among other purposes, the FBI used its illegally obtained information to alert prosecuting attorneys about the planned legal strategies of NLG defense lawyers.[citation needed][26] The FBI also used illegal undercover operations to disrupt Communist and other dissident political groups. In 1956, Hoover was becoming increasingly frustrated by Supreme Court decisions that limited the Justice Department's ability to prosecute Communists. At this time, he formalized a covert "dirty tricks" program under the name COINTELPRO.[24] COINTELPRO actions included planting forged documents to create the suspicion that a key person was an FBI informer, spreading rumors through anonymous letters, leaking information to the press, calling for IRS audits, and the like. The COINTELPRO program remained in operation until 1971. Historian Ellen Schrecker calls the FBI "the single most important component of the anti-communist crusade" and writes: "Had observers known in the 1950s what they have learned since the 1970s, when the Freedom of Information Act opened the Bureau's files, 'McCarthyism' would probably be called 'Hooverism'."[27] Congress [ edit ] House Committee on Un-American Activities [ edit ] The House Committee on Un-American Activities – commonly referred to as the HUAC – was the most prominent and active government committee involved in anti-communist investigations. Formed in 1938 and known as the Dies Committee, for Rep. Martin Dies, who chaired it until 1944, HUAC investigated a variety of "activities", including those of German-American Nazis during World War II. The committee soon focused on Communism, beginning with an investigation into Communists in the Federal Theatre Project in 1938. A significant step for HUAC was its investigation of the charges of espionage brought against Alger Hiss in 1948. This investigation ultimately resulted in Hiss's trial and conviction for perjury, and convinced many of the usefulness of congressional committees for uncovering Communist subversion. HUAC achieved its greatest fame and notoriety with its investigation into the Hollywood film industry. In October 1947, the Committee began to subpoena screenwriters, directors, and other movie-industry professionals to testify about their known or suspected membership in the Communist Party, association with its members, or support of its beliefs. At these testimonies, what became known as "the $64,000 question" was asked: "Are you now or have you ever been a member of the Communist Party of the United States?"[28] Among the first film industry witnesses subpoenaed by the committee were ten who decided not to cooperate. These men, who became known as the "Hollywood Ten", cited the First Amendment's guarantee of free speech and free assembly, which they believed legally protected them from being required to answer the committee's questions. This tactic failed, and the ten were sentenced to prison for contempt of Congress. Two of them were sentenced to six months, the rest to a year. In the future, witnesses (in the entertainment industries and otherwise) who were determined not to cooperate with the committee would claim their Fifth Amendment protection against self-incrimination. While this usually protected them from a contempt-of-Congress citation, it was considered grounds for dismissal by many government and private-industry employers. The legal requirements for Fifth Amendment protection were such that a person could not testify about his own association with the Communist Party and then refuse to "name names" of colleagues with Communist affiliations.[29] Thus, many faced a choice between "crawl[ing] through the mud to be an informer," as actor Larry Parks put it, or becoming known as a "Fifth Amendment Communist"—an epithet often used by Senator McCarthy.[30] Senate committees [ edit ] In the Senate, the primary committee for investigating Communists was the Senate Internal Security Subcommittee (SISS), formed in 1950 and charged with ensuring the enforcement of laws relating to "espionage, sabotage, and the protection of the internal security of the United States." The SISS was headed by Democrat Pat McCarran and gained a reputation for careful and extensive investigations. This committee spent a year investigating Owen Lattimore and other members of the Institute of Pacific Relations. As had been done numerous times before, the collection of scholars and diplomats associated with Lattimore (the so-called China Hands) were accused of "losing China", and while some evidence of pro-communist attitudes was found, nothing supported McCarran's accusation that Lattimore was "a conscious and articulate instrument of the Soviet conspiracy". Lattimore was charged with perjuring himself before the SISS in 1952. After many of the charges were rejected by a federal judge and one of the witnesses confessed to perjury, the case was dropped in 1955.[31] McCarthy headed the Senate Permanent Subcommittee on Investigations in 1953 and 1954, and during that time, used it for a number of his Communist-hunting investigations. McCarthy first examined allegations of Communist influence in the Voice of America, and then turned to the overseas library program of the State Department. Card catalogs of these libraries were searched for works by authors McCarthy deemed inappropriate. McCarthy then recited the list of supposedly pro-communist authors before his subcommittee and the press. Yielding to the pressure, the State Department ordered its overseas librarians to remove from their shelves "material by any controversial persons, Communists, fellow travelers, etc." Some libraries actually burned the newly forbidden books.[32] McCarthy's committee then began an investigation into the United States Army. This began at the Army Signal Corps laboratory at Fort Monmouth. McCarthy garnered some headlines with stories of a dangerous spy ring among the Army researchers, but ultimately nothing came of this investigation.[33] McCarthy next turned his attention to the case of a U.S. Army dentist who had been promoted to the rank of major despite having refused to answer questions on an Army loyalty review form. McCarthy's handling of this investigation, including a series of insults directed at a brigadier general, led to the Army–McCarthy hearings, with the Army and McCarthy trading charges and counter-charges for 36 days before a nationwide television audience. While the official outcome of the hearings was inconclusive, this exposure of McCarthy to the American public resulted in a sharp decline in his popularity.[34] In less than a year, McCarthy was censured by the Senate, and his position as a prominent force in anti-communism was essentially ended.[35] Blacklists [ edit ] On November 25, 1947, the day after the House of Representatives approved citations of contempt for the Hollywood Ten, Eric Johnston, president of the Motion Picture Association of America, issued a press release on behalf of the heads of the major studios that came to be referred to as the Waldorf Statement. This statement announced the firing of the Hollywood Ten and stated: "We will not knowingly employ a Communist or a member of any party or group which advocates the overthrow of the government of the United States..." This marked the beginning of the Hollywood blacklist. In spite of the fact that hundreds would be denied employment, the studios, producers, and other employers did not publicly admit that a blacklist existed. At this time, private loyalty-review boards and anti-communist investigators began to appear to fill a growing demand among certain industries to certify that their employees were above reproach. Companies that were concerned about the sensitivity of their business, or which, like the entertainment industry, felt particularly vulnerable to public opinion made use of these private services. For a fee, these teams would investigate employees and question them about their politics and affiliations. At such hearings, the subject would usually not have a right to the presence of an attorney, and as with HUAC, the interviewee might be asked to defend himself against accusations without being allowed to cross-examine the accuser. These agencies would keep cross-referenced lists of leftist organizations, publications, rallies, charities, and the like, as well as lists of individuals who were known or suspected communists. Books such as Red Channels and newsletters such as Counterattack and Confidential Information were published to keep track of communist and leftist organizations and individuals.[36] Insofar as the various blacklists of McCarthyism were actual physical lists, they were created and maintained by these private organizations.[citation needed][further explanation needed] Laws and arrests [ edit ] Efforts to protect the United States from the perceived threat of Communist subversion were particularly enabled by several federal laws. The Alien Registration Act or Smith Act of 1940 made the act of "knowingly or willfully advocate, abet, advise or teach the ... desirability or propriety of overthrowing the Government of the United States or of any State by force or violence, or for anyone to organize any association which teaches, advises or encourages such an overthrow, or for anyone to become a member of or to affiliate with any such association" a criminal offense. Hundreds of Communists and others were prosecuted under this law between 1941 and 1957. Eleven leaders of the Communist Party were convicted under the Smith Act in 1949 in the Foley Square trial. Ten defendants were given sentences of five years and the eleventh was sentenced to three years. The defense attorneys were cited for contempt of court and given prison sentences.[37] In 1951, 23 other leaders of the party were indicted, including Elizabeth Gurley Flynn, a founding member of the American Civil Liberties Union. Many were convicted on the basis of testimony that was later admitted to be false.[38] By 1957, 140 leaders and members of the Communist Party had been charged under the law, of whom 93 were convicted.[39] The McCarran Internal Security Act, which became law in 1950, has been described by scholar Ellen Schrecker as "the McCarthy era's only important piece of legislation"[40] (the Smith Act technically antedated McCarthyism). However, the McCarran Act had no real effect beyond legal harassment. It required the registration of Communist organizations with the U.S. Attorney General and established the Subversive Activities Control Board to investigate possible Communist-action and Communist-front organizations so they could be required to register. Due to numerous hearings, delays, and appeals, the act was never enforced, even with regard to the Communist Party of the United States itself, and the major provisions of the act were found to be unconstitutional in 1965 and 1967.[41] In 1952, the Immigration and Nationality, or McCarran–Walter, Act was passed. This law allowed the government to deport immigrants or naturalized citizens engaged in subversive activities and also to bar suspected subversives from entering the country. The Communist Control Act of 1954 was passed with overwhelming support in both houses of Congress after very little debate. Jointly drafted by Republican John Marshall Butler and Democrat Hubert Humphrey, the law was an extension of the Internal Security Act of 1950, and sought to outlaw the Communist Party by declaring that the party, as well as "Communist-Infiltrated Organizations" were "not entitled to any of the rights, privileges, and immunities attendant upon legal bodies." While the Communist Control Act had an odd mix of liberals and conservatives among its supporters, it never had any significant effect. The act was successfully applied only twice; in 1954 it was used to prevent Communist Party members from appearing on the New Jersey state ballot, and in 1960, it was cited to deny the CPUSA recognition as an employer under New York state's unemployment compensation system. The New York Post called the act "a monstrosity", "a wretched repudiation of democratic principles," while The Nation accused Democratic liberals of a "neurotic, election-year anxiety to escape the charge of being 'soft on Communism' even at the expense of sacrificing constitutional rights."[42] Repression in the individual states [ edit ] In addition to the federal laws and responding to the worries of the local opinion, several states enacted anti-communist statutes. By 1952, several states had enacted statutes against criminal anarchy, criminal syndicalism, and sedition; banned from public employment or even from receiving public aid, Communists and "subversives"; asked for loyalty oaths from public servants, and severely restricted or even banned the Communist party. In addition, six states, among them California[43] (see California Senate Factfinding Subcommittee on Un-American Activities) had equivalents to the HUAC.[44] Some of these states had very severe, or even extreme, laws against Communism. In 1950, Michigan enacted life imprisonment for subversive propaganda; the following year, Tennessee enacted death penalty for advocating the violent overthrow of the government.[44] Death penalty for membership of the Communist party was discussed in Texas by Governor Allan Shivers, who described it as "worse than murder."[45][46] Municipalities and counties also enacted anti-communist ordinances: Los Angeles banned any Communist or "Muscovite model of police-state dictatorship" from owning any arm and Birmingham, Alabama, and Jacksonville, Florida, banned any Communist from being within the city's limits.[44] Popular support [ edit ] Flier issued in May 1955 by the Keep America Committee urging readers to "fight communistic world government" by opposing public health programs McCarthyism was supported by a variety of groups, including the American Legion and various other anti-communist organizations. One core element of support was a variety of militantly anti-communist women's groups such as the American Public Relations Forum and the Minute Women of the U.S.A.. These organized tens of thousands of housewives into study groups, letter-writing networks, and patriotic clubs that coordinated efforts to identify and eradicate what they saw as subversion.[47] Although far-right radicals were the bedrock of support for McCarthyism, they were not alone. A broad "coalition of the aggrieved" found McCarthyism attractive, or at least politically useful. Common themes uniting the coalition were opposition to internationalism, particularly the United Nations; opposition to social welfare provisions, particularly the various programs established by the New Deal; and opposition to efforts to reduce inequalities in the social structure of the United States.[48] One focus of popular McCarthyism concerned the provision of public-health services, particularly vaccination, mental health care services, and fluoridation, all of which were denounced by some to be communist plots to poison or brainwash the American people. Such viewpoints led to collisions between McCarthyite radicals and supporters of public-health programs, most notably in the case of the Alaska Mental Health Bill controversy of 1956.[49] William F. Buckley Jr., the founder of the influential conservative political magazine National Review, wrote a defense of McCarthy, McCarthy and his Enemies, in which he asserted that "McCarthyism ... is a movement around which men of good will and stern morality can close ranks."[50] In addition, as Richard Rovere points out, many ordinary Americans became convinced that there must be "no smoke without fire" and lent their support to McCarthyism. The Gallup poll found that at his peak in January 1954, 50% of the American public supported McCarthy, while 29% had an unfavorable opinion. His support fell to 34% in June 1954.[51] Republicans tended to like what McCarthy was doing and Democrats did not, and McCarthy also had significant support from traditional Democratic ethnic groups, especially Catholics, as well as many unskilled workers and small-business owners. (McCarthy himself was a Catholic.) He had very little support among union activists and Jews.[52] Portrayals of Communists [ edit ] Those who sought to justify McCarthyism did so largely through their characterization of Communism, and American Communists in particular. Proponents of McCarthyism claimed that the CPUSA was so completely under Moscow's control that any American Communist was a puppet of the Soviet intelligence services. This view is supported by recent documentation from the archives of the KGB[53] as well as post-war decodes of wartime Soviet radio traffic from the Venona Project,[54] showing that Moscow provided financial support to the CPUSA and had significant influence on CPUSA policies. J. Edgar Hoover commented in a 1950 speech, "Communist members, body and soul, are the property of the Party." This attitude was not confined to arch-conservatives. In 1940, the American Civil Liberties Union ejected founding member Elizabeth Gurley Flynn, saying that her membership in the Communist Party was enough to disqualify her as a civil libertarian. In the government's prosecutions of Communist Party members under the Smith Act (see above), the prosecution case was based not on specific actions or statements by the defendants, but on the premise that a commitment to violent overthrow of the government was inherent in the doctrines of Marxism–Leninism. Passages of the CPUSA's constitution that specifically rejected revolutionary violence were dismissed as deliberate deception.[55] In addition, the party was often claimed to not allow any member to resign, so a person who had been a member for a short time decades previously could be considered as suspect as a current member. Many of the hearings and trials of McCarthyism featured testimony by former Communist Party members such as Elizabeth Bentley, Louis Budenz, and Whittaker Chambers, speaking as expert witnesses.[56][57] Various historians and pundits have discussed alleged Soviet-directed infiltration of the U.S. government and the possible collaboration of high U.S. government officials.[58][59][60][61] Victims of McCarthyism [ edit ] Estimating the number of victims of McCarthy is difficult. The number imprisoned is in the hundreds, and some ten or twelve thousand lost their jobs.[62] In many cases, simply being subpoenaed by HUAC or one of the other committees was sufficient cause to be fired.[63] Many of those who were imprisoned, lost their jobs, or were questioned by committees did, in fact, have a past or present connection of some kind with the Communist Party. For the vast majority, though, both the potential for them to do harm to the nation and the nature of their communist affiliation were tenuous.[64] After the extremely damaging "Cambridge Five" spy scandal (Guy Burgess, Donald Maclean, Kim Philby, Anthony Blunt, et al.), suspected homosexuality was also a common cause for being targeted by McCarthyism. The hunt for "sexual perverts", who were presumed to be subversive by nature, resulted in over 5,000 federal workers being fired, and thousands were harassed and denied employment.[65][66] Many have termed this aspect of McCarthyism the "lavender scare".[67][68] Homosexuality was classified as a psychiatric disorder in the 1950s.[69] However, in the context of the highly politicized Cold War environment, homosexuality became framed as a dangerous, contagious social disease that posed a potential threat to state security.[69] As the family was believed to be the cornerstone of American strength and integrity,[70] the description of homosexuals as "sexual perverts" meant that they were both unable to function within a family unit and presented the potential to poison the social body.[71] This era also witnessed the establishment of widely spread FBI surveillance intended to identify homosexual government employees.[72] The McCarthy hearings and according "sexual pervert" investigations can be seen to have been driven by a desire to identify individuals whose ability to function as loyal citizens had been compromised.[71] McCarthy began his campaign by drawing upon the ways in which he embodied traditional American values to become the self-appointed vanguard of social morality.[73] Accusations of alleged homosexual behavior marked the end of McCarthy's political career.[74] In the film industry, more than 300 actors, authors, and directors were denied work in the U.S. through the unofficial Hollywood blacklist. Blacklists were at work throughout the entertainment industry, in universities and schools at all levels, in the legal profession, and in many other fields. A port-security program initiated by the Coast Guard shortly after the start of the Korean War required a review of every maritime worker who loaded or worked aboard any American ship, regardless of cargo or destination. As with other loyalty-security reviews of McCarthyism, the identities of any accusers and even the nature of any accusations were typically kept secret from the accused. Nearly 3,000 seamen and longshoremen lost their jobs due to this program alone.[75] Some of the notable people who were blacklisted or suffered some other persecution during McCarthyism include: In 1953, Robert K. Murray, a young professor of history at Pennsylvania State University who had served as an intelligence officer in World War II, was revising his dissertation on the Red Scare of 1919–20 for publication until Little, Brown and Company decided that "under the circumstances ... it wasn't wise for them to bring this book out." He learned that investigators were questioning his colleagues and relatives. The University of Minnesota press published his volume, Red Scare: A Study in National Hysteria, 1919–1920, in 1955.[124] Critical reactions [ edit ] The nation was by no means united behind the policies and activities that have come to be associated with McCarthyism. The many critics of various aspects of McCarthyism included many figures not generally noted for their liberalism. For example, in his overridden veto of the McCarran Internal Security Act of 1950, President Truman wrote, "In a free country, we punish men for the crimes they commit, but never for the opinions they have."[125] Truman also unsuccessfully vetoed the Taft–Hartley Act, which among other provisions denied trade unions National Labor Relations Board protection unless union leaders signed affidavits swearing they were not and had never been Communists. In 1953, after he left office, Truman criticized the current Eisenhower administration: It is now evident that the present Administration has fully embraced, for political advantage, McCarthyism. I am not referring to the Senator from Wisconsin. He is only important in that his name has taken on the dictionary meaning of the word. It is the corruption of truth, the abandonment of the due process law. It is the use of the big lie and the unfounded accusation against any citizen in the name of Americanism or security. It is the rise to power of the demagogue who lives on untruth; it is the spreading of fear and the destruction of faith in every level of society.[126] On June 1, 1950, Senator Margaret Chase Smith, a Maine Republican, delivered a speech to the Senate she called a "Declaration of Conscience". In a clear attack upon McCarthyism, she called for an end to "character assassinations" and named "some of the basic principles of Americanism: The right to criticize; The right to hold unpopular beliefs; The right to protest; The right of independent thought". She said "freedom of speech is not what it used to be in America", and decried "cancerous tentacles of 'know nothing, suspect everything' attitudes".[127] Six other Republican senators—Wayne Morse, Irving M. Ives, Charles W. Tobey, Edward John Thye, George Aiken, and Robert C. Hendrickson—joined Smith in condemning the tactics of McCarthyism. Elmer Davis, one of the most highly respected news reporters and commentators of the 1940s and 1950s, often spoke out against what he saw as the excesses of McCarthyism. On one occasion he warned that many local anti-communist movements constituted a "general attack not only on schools and colleges and libraries, on teachers and textbooks, but on all people who think and write ... in short, on the freedom of the mind".[128] In 1952, the Supreme Court upheld a lower-court decision in Adler v. Board of Education of New York, thus approving a law that allowed state loyalty review boards to fire teachers deemed "subversive". In his dissenting opinion, Justice William O. Douglas wrote: "The present law proceeds on a principle repugnant to our society—guilt by association.... What happens under this law is typical of what happens in a police state. Teachers are under constant surveillance; their pasts are combed for signs of disloyalty; their utterances are watched for clues to dangerous thoughts."[129] One of the most influential opponents of McCarthyism was the famed CBS newscaster and analyst Edward R. Murrow. On October 20, 1953, Murrow's show See It Now aired an episode about the dismissal of Milo Radulovich, a former reserve Air Force lieutenant who was accused of associating with Communists. The show was strongly critical of the Air Force's methods, which included presenting evidence in a sealed envelope that Radulovich and his attorney were not allowed to open. On March 9, 1954, See It Now aired another episode on the issue of McCarthyism, this one attacking Joseph McCarthy himself. Titled "A Report on Senator Joseph R. McCarthy", it used footage of McCarthy speeches to portray him as dishonest, reckless, and abusive toward witnesses and prominent Americans. In his concluding comment, Murrow said: We must not confuse dissent with disloyalty. We must remember always that accusation is not proof and that conviction depends upon evidence and due process of law. We will not walk in fear, one of another. We will not be driven by fear into an age of unreason, if we dig deep in our history and our doctrine, and remember that we are not descended from fearful men.[30] This broadcast has been cited as a key episode in bringing about the end of McCarthyism.[130] In April 1954, McCarthy was also under attack in the Army–McCarthy hearings. These hearings were televised live on the new American Broadcasting Company network, allowing the public to view first-hand McCarthy's interrogation of individuals and his controversial tactics. In one exchange, McCarthy reminded the attorney for the Army, Joseph Welch, that he had an employee in his law firm who had belonged to an organization that had been accused of Communist sympathies. In an exchange that reflected the increasingly negative public opinion of McCarthy, Welch rebuked the senator: "Have you no sense of decency, sir? At long last, have you left no sense of decency?"[131] Decline [ edit ] In the mid and late 1950s, the attitudes and institutions of McCarthyism slowly weakened. Changing public sentiments heavily contributed to the decline of McCarthyism. Its decline may also be charted through a series of court decisions. A key figure in the end of the blacklisting of McCarthyism was John Henry Faulk. Host of an afternoon comedy radio show, Faulk was a leftist active in his union, the American Federation of Television and Radio Artists. He was scrutinized by AWARE, Inc., one of the private firms that examined individuals for signs of communist "disloyalty". Marked by AWARE as unfit, he was fired by CBS Radio. Almost uniquely among the many victims of blacklisting, Faulk decided to sue AWARE in 1957 and finally won the case in 1962.[132] With this court decision, the private blacklisters and those who used them were put on notice that they were legally liable for the professional and financial damage they caused. Although some informal blacklisting continued, the private "loyalty checking" agencies were soon a thing of the past.[133] Even before the Faulk verdict, many in Hollywood had decided it was time to break the blacklist. In 1960, Dalton Trumbo, one of the best known members of the Hollywood Ten, was publicly credited with writing the films Exodus and Spartacus. Much of the undoing of McCarthyism came at the hands of the Supreme Court. As Richard Rovere wrote in his biography of Joseph McCarthy, "[T]he United States Supreme Court took judicial notice of the rents McCarthy was making in the fabric of liberty and thereupon wrote a series of decisions that have made the fabric stronger than before."[134] Two Eisenhower appointees to the court—Earl Warren (who was made Chief Justice) and William J. Brennan, Jr.—proved to be more liberal than Eisenhower had anticipated, and he would later refer to the appointment of Warren as his "biggest mistake".[135] In 1956, the Supreme Court heard the case of Slochower v. Board of Education. Harry Slochower was a professor at Brooklyn College who had been fired by New York City for invoking the Fifth Amendment when McCarthy's committee questioned him about his past membership in the Communist Party. The court prohibited such actions, ruling "...we must condemn the practice of imputing a sinister meaning to the exercise of a person's constitutional right under the Fifth Amendment.... The privilege against self-incrimination would be reduced to a hollow mockery if its exercise could be taken as equivalent either to a confession of guilt or a conclusive presumption of perjury."[136] The 1956 Cole v. Young ruling also greatly weakened the ability to discriminate in the federal civilian workforce.[137] Another key decision was in the 1957 case Yates v. United States, in which the convictions of fourteen Communists were reversed. In Justice Black's opinion, he wrote of the original "Smith Act" trials: "The testimony of witnesses is comparatively insignificant. Guilt or innocence may turn on what Marx or Engels or someone else wrote or advocated as much as a hundred years or more ago.... When the propriety of obnoxious or unfamiliar view about government is in reality made the crucial issue, ... prejudice makes conviction inevitable except in the rarest circumstances."[138] Also in 1957, the Supreme Court ruled on the case of Watkins v. United States, curtailing the power of HUAC to punish uncooperative witnesses by finding them in contempt of Congress. Justice Warren wrote in the decision: "The mere summoning of a witness and compelling him to testify, against his will, about his beliefs, expressions or associations is a measure of governmental interference. And when those forced revelations concern matters that are unorthodox, unpopular, or even hateful to the general public, the reaction in the life of the witness may be disastrous."[139][140] In its 1958 decision in Kent v. Dulles, the Supreme Court halted the State Department from using the authority of its own regulations to refuse or revoke passports based on an applicant's communist beliefs or associations.[141] Repercussions [ edit ] The political divisions McCarthyism created in the United States continue to make themselves manifest, and the politics and history of a anti-communism in the United States are still contentious. Portions of the massive security apparatus established during the McCarthy era still exist. Loyalty oaths are still required by the California Constitution for all officials and employees of the government of California (which is highly problematic for Quakers and Jehovah's Witnesses whose beliefs preclude them from pledging absolute loyalty to the state).[142] At the federal level, a few portions of the McCarran Internal Security Act remain in effect. A number of observers have compared the oppression of liberals and leftists during the McCarthy period to recent actions against suspected terrorists, most of them Muslims. In The Age of Anxiety: McCarthyism to Terrorism, author Haynes Johnson compares the "abuses suffered by aliens thrown into high-security U.S. prisons in the wake of 9/11" to the excesses of the McCarthy era.[143] Similarly, David D. Cole has written that the Patriot Act "in effect resurrects the philosophy of McCarthyism, simply substituting 'terrorist' for 'communist'".[144] From the opposite pole, conservative writer Ann Coulter devotes much of her book Treason to drawing parallels between past opposition to McCarthy and McCarthyism and the policies and beliefs of modern-day liberals, arguing that the former hindered the anti-communist cause and the latter hinder the War on Terrorism.[145] Other authors who have drawn on a comparison between current anti-terrorism policies and McCarthyism include Geoffrey R. Stone,[146] Ted Morgan,[147] and Jonah Goldberg.[148] McCarthyism also attracts controversy purely as a historical issue. Through declassified documents from Soviet archives and Venona project decryptions of coded Soviet messages, the Soviet Union was found to have engaged in substantial espionage activities in the United States during the 1940s. The Communist Party USA also was substantially funded and its policies controlled by the Soviet Union, and accusations existed that CPUSA members were often recruited as spies.[149] In the view of some contemporary commentators, these revelations stand as at least a partial vindication of McCarthyism.[150] Some feel that a genuinely dangerous subversive element was in the United States, and that this danger justified extreme measures.[148] Others, while acknowledging that inexcusable excesses occurred during McCarthyism, argue that some contemporary historians of McCarthyism underplay the depth of Soviet espionage in America[151] or the undemocratic nature of the CPUSA,[152] the latter concern being shared by some Trotskyites who felt that they, and anti-Stalin socialists in general, were persecuted by the CPUSA.[153] The opposing view holds that, recent revelations notwithstanding, by the time McCarthyism began in the late 1940s, the CPUSA was an ineffectual fringe group, and the damage done to U.S. interests by Soviet spies after World War II was minimal.[154] Historian Ellen Schrecker, herself criticised for pro-Stalinist leanings,[155] has written, "in this country, McCarthyism did more damage to the constitution than the American Communist Party ever did."[156] Later use of the term [ edit ] Since the time of McCarthy, the word McCarthyism has entered American speech as a general term for a variety of practices: aggressively questioning a person's patriotism, making poorly supported accusations, using accusations of disloyalty to pressure a person to adhere to conformist politics or to discredit an opponent, subverting civil and political rights in the name of national security, and the use of demagoguery are all often referred to as McCarthyism.[157][158][159] McCarthyism can also be synonymous with the term witch-hunt, both referring to mass hysteria and moral panic.[160] In popular culture [ edit ] The 1951 novel The Troubled Air by Irwin Shaw tells the story of the director of a (fictional) radio show, broadcast live at the time, who is given a deadline to investigate his cast for alleged links to communism. The novel recounts the devastating effects on all concerned.[161] The 1952 Arthur Miller play The Crucible used the Salem witch trials as a metaphor for McCarthyism, suggesting that the process of McCarthyism-style persecution can occur at any time or place. The play focused on the fact that once accused, a person had little chance of exoneration, given the irrational and circular reasoning of both the courts and the public. Miller later wrote: "The more I read into the Salem panic, the more it touched off corresponding images of common experiences in the fifties."[162] The 1976 film The Front starring Woody Allen dealt with the McCarthy-era Hollywood blacklist. The film was made by those blacklisted: producer and director Martin Ritt; writer Walter Bernstein; and actors Zero Mostel, Herschel Bernardi, Michael Murphy, John Randolph, Lloyd Gough, and Joshua Shelley.[163] Guilty by Suspicion is a 1991 American drama film about the Hollywood blacklist, McCarthyism, and the activities of the HUAC. Written and directed by Irwin Winkler, it starred Robert De Niro, Annette Bening, and George Wendt. The 2005 film Good Night, and Good Luck by George Clooney starred David Strathairn as broadcast journalist Edward R. Murrow and contained archival footage of McCarthy.[164] See also [ edit ] References [ edit ] Citations [ edit ] Sources [ edit ] Historiography [ edit ] Haynes, John Earl. "The Cold War debate continues: A traditionalist view of historical writing on domestic Communism and anti-Communism." Journal of Cold War Studies 2.1 (2000): 76-115. 2.1 (2000): 76-115. Hixson Jr, William B. Search for the American right wing: An analysis of the social science record, 1955–1987 (Princeton University Press, 2015). (Princeton University Press, 2015). Reeves, Thomas C. "McCarthyism: Interpretations since Hofstadter." Wisconsin Magazine of History (1976): 42–54. online (1976): 42–54. online Selverstone, Marc J. "A Literature So Immense: The Historiography of Anticommunism." Organization of American Historians Magazine of History 24.4 (2010): 7–11.

Ten Steps to Create Virtual Smile Design Templates With Adobe Photoshop® CS6. Computer design software has become a primary tool for communication among the dentist, patient, and ceramist. Virtual smile design can be carried out using various software programs, most of which use assorted forms of teeth templates that are made based on the concept of "golden proportion." Despite current advances in 3-dimensional imaging and smile designing, many clinicians still employ conventional design methods and analog (ie, man-made) mock-ups in assessing and establishing esthetic makeovers. To simplify virtual smile designing, the teeth templates should be readily available. No literature has provided details as to how to create these templates. This article explains a technique for creating different forms of teeth templates using Adobe Photoshop® CS6 that eventually can be used for smile design purposes, either in Photoshop or Microsoft Powerpoint. Clinically speaking, various smile design templates created using set proportions in Adobe Photoshop CS6 can be used in virtual smile designing, a valuable resource in diagnosis, treatment planning, and communicating with patients and ceramists, thus providing a platform for a successful esthetic rehabilitation.

Bacterial Cellulose Membranes as Carriers for Nisin: Incorporation, Antimicrobial Activity, Cytotoxicity and Morphology Based on the previous study, in which nisin and bacterial cellulose were utilized, this new experiment loads nisin into bacterial cellulose (NBC) and evaluates the morphological characteristics, cytotoxicity, antimicrobial activity and stability of the developed system. The load efficiency of nisin in BC was evaluated by an agar diffusion assay, utilizing Lactobacillus sakei, and total proteins. After having found the ideal time and concentration for the loading process, the system stability was evaluated for 100 days at 4, 25 and 37 °C against Staphylococcus aureus and L. sakei. Thus, in this study, there is a system that proves to be efficient, once BC has enhanced the antimicrobial activity of nisin, acting as a selective barrier for other compounds present in the standard solution and protecting the peptide. After 4 h, with 45% of proteins, this activity was almost 2 log10 higher than that of the initial solution. Once the nisin solution was not pure, it is possible to suggest that the BC may have acted as a filter. This barrier enhanced the nisin activity and, as a consequence of the nisin loading, a stable NBC system formed. The NBC could create meaningful material for pharmaceutical and food applications. Introduction Antimicrobial peptides are synthesized by several microorganisms. Nisin is a lantibiotic peptide, with 34 amino acids, and is secreted by Lactococcus. The antimicrobial property of nisin is attributed to pore formation in the cell membrane of microorganisms, with specific binding to the lipidic precursor of the cell wall, attached to the membrane. Nisin is considered safe by the World Health Organization (WHO) and the Food and Drug Administration (FDA-United States), being used initially as a food additive. Further, its antimicrobial action encourages the clinical use of nisin, whether in topical or systemic therapies, due to its broad-spectrum activity and lower probability of developing microbial resistance. The application of nisin extends to several medical areas, from mastitis to oral and gastrointestinal diseases. Bacterial cellulose (BC) is a polysaccharide that is extracellularly secreted by several microorganisms, such as Agrobacterium, Rhizobium, Escherichia, Sarcina, and Acetobacter. Namely, Komagataeibacter xylinus is a non-pathogenic Gram-negative bacterium that can produce significant amounts of cellulose. BC is a linear glucose polymer, formed by a matrix of nanofibers, giving it porous characteristics in a three-dimensional network structure. Although its structure resembles vegetable cellulose, bacterial cellulose presents a high degree of purity, crystallinity, tensile strength, and high-water absorption. Due to its biocompatibility and non-toxicity, the BC application has been directed to medical devices and tissue engineering. Additionally, owing to its structural model, BC has been proposed as an ideal dermal substitute that is capable of inducing the direction of cells for repair in tissue reconstruction. BC has been used as wound dressing because it forms a physical barrier against infections. It also allows gas exchange, absorbs exudates, keeps the wound moist to favor tissue reconstitution and it can be easily removed. The incorporation of biomolecules into BC has been studied both by increasing its antibacterial or enzymatic properties and by providing a control release system. For instance, nisin immobilization in solid matrices such as bacterial cellulose membranes could control its release. Due to these properties, our research group previously evaluated the antimicrobial and antioxidant activity of nisin loaded into BC. Since the work yielded satisfactory results, our group has provided a supplementary study. For this reason, this study has evaluated BC membranes as carriers for nisin regarding the morphological characteristics, cytotoxicity, and stability of the developed system. Materials The standard nisin and the bicinchoninic acid kit were purchased from Sigma-Aldrich (So Paulo, Brazil). All other reagents were of analytical grade. BC Production and Purification BC was produced by Komagataeibacter xylinus ATCC 53582, using 24-well plates with Hestrin and Schramm medium. Each well was filled with 1 mL and incubated at 30 C in static conditions for seven days. After that, membranes were immersed in a 2% sulfate dodecyl sodium (SDS) solution under stirring overnight and washed in running water. The bleaching process was carried out with 4% NaOH for one hour until reaching 60 C. The BC membranes were washed in order to remove NaOH and sterilized at 121 C for 15 min. Total Proteins Quantification Protein concentration was determined by a bicinchoninic acid assay. Bovine serum albumin (BSA) with different concentrations from 0.1 to 1.0 mg/mL was used as the standard protein. Absorbance reading was performed on 96-well microplates with a wavelength of 562 nm, by spectroscopy (Infinite M200 PRO, RCHISTO, Barueri, Brazil). Analyses were performed in triplicate, and the mean of these absorbance values was used to determine protein concentrations. Nisin Standard Curve by Agar Diffusion Assay The standard nisin (Sigma-Aldrich, St. Louis, MO, USA-containing 2.5% nisin, with 1,000,000 IU/g in its composition) solution was prepared by dissolving 1 g of nisin in 10 mL of phosphate buffer solution (PBS-pH 7.0), as in Table 1. The solution was centrifuged at 13,201 g for 10 min at 10 C and the supernatant was collected and filtered in a 0.22 m membrane (Millipore, Burlington, MA, USA). The nisin standard curve was evaluated by an agar diffusion assay. The nisin bioindicator Lactobacillus sakei ATCC 15521 was used for the agar diffusion assay. The concentrations of standard nisin were related by the diameter of the inhibition halo (H, mm), and the activity of nisin was determined and expressed in arbitrary units per mL (AU/mL). The activity of nisin was based on the dilution of the standard nisin calibration curves. The correlation between AU/mL and international units per mL (IU/mL) was 1.09 ± 0.17 AU to 1.0 IU (40 IU = 1 g of pure nisin A). Evaluation of Nisin Loaded into BC by Time A dose of 250 g/mL of nisin was employed based on previous studies from our research group. The BC membranes were arranged in 24-well plates and 1.0 mL of nisin solution (250 g/mL) was added in each well. The plate was placed on a shaker (NT 715, Nova Tecnica) at 25 C, under 100 rpm for 4, 8, 12, 18 and 24 h. After each period, residual samples were collected in order to determine the nisin load efficiency by the concentration of proteins and nisin activity. The BC membranes were collected and evaluated by the agar diffusion assay for the measurement of the nisin activity. The residual samples were evaluated by the agar diffusion assay and the total protein method. The load efficiency (LE) of nisin was evaluated by the total protein method and calculated by the following Equation, which was elaborated by the authors: %LE = Protein in the initial solution − Protein in the Residual Solution Protein in the initial solution 100 Stability Test by Agar Difusion Assay In this test, Staphylococcus aureus ATCC 10390 and Lactobacillus sakei ATCC 15521 were utilized. S. aureus was grown in Tryptic Soy Broth (TSB) supplied by Becton, Dickinson and Company (BD Difco TM, Franklin Lakes, NJ, USA) for 24 h at 37 C, whereas L. sakei was cultivated in De Man, Rogosa and Sharpe broth (MRS) supplied by Sigma-Aldrich Corporation (Sigma-Aldrich, St. Louis, MO, USA) for 24 h at 30 C. All the media were prepared in distilled water and autoclaved at 121 C for 30 min. After the growth, the microorganisms were counted by the Pour Plate technique and the sample with 10 6 UFC/mL was selected to carry out the agar diffusion assay. For S. aureus, an amount of 20 mL of TSB agar was placed in a Petri plate. After the solidification, 1 mL was placed on the agar surface. For L. sakei, an amount of 20 mL of MRS agar was placed in a Petri plate. After the solidification, 1 mL (10 6 UFC/mL) was also placed on the agar surface. The BC membranes loaded with nisin (N-BC) were kept in Petri plates and incubated at different temperatures (4, 25, 37 C)-pre-set temperature conditions for stability evaluation-for 100 days. The samples were collected each day and the nisin activity was evaluated by the agar diffusion assay, adapted from ISO 20645, with L. sakei and S. aureus. The BC was placed on an agar surface and the possible clear zones were observed after its incubation at 37 C for 24 h. The presence of a clear halo that formed around BC was measured and indicated the antimicrobial activity. Cytotoxicity Assay The standard nisin and N-BC at the best time and with different concentrations of 7, 15, 31, 62, 125, 250 g/mL were arranged in 24-well plates with 500 L of DMEM (Dulbecco's modified eagle's medium, Low Glucose, Sigma ® ), with the addition of 10% fetal bovine serum (FBS, Sigma ® ) and 1% antibiotic (PEN/STREP, Sigma ® ) in each well. In each well, there were 75,000 L929 fibroblast cells. The plates were incubated at 37 C in an atmosphere of 5% CO2 for 24 and 48 h. The assay was performed in triplicate and for positive control, PBS and DMEM were used. After 24 and 48 h, cell viability was determined by the mitochondrial activity of the cell culture by the colorimetric assay MTS . Absorbance readings were taken at 490 nm, after 1 h of incubation of the cells with the reagent. The results are expressed as mean percentage ± standard deviation (SD) of viable cells in relation to the positive control. The BC and N-BC (with 250 /mL of nisin) were stored at −80 C for 24 h and lyophilized for 48 h. BC and N-BC were set with carbon tape and metallized for 2 min (DH-29010SCTR Smart Coater). SEM images were observed on the scanning electron microscopy equipment (JEOL, Tokyo, Japan, Model IT200) and obtained using an accelerating voltage of 20 kV. X-ray Microtomography (CT) This equipment is responsible for analyzing pore size, porosity (%), and the interconnectivity of the pores in the membranes. The membrane pictures were captured by X-ray microtomography (Bruckermicro CT-SkyScan 1174, Kontich, Belgium). The X-ray source was 29 kV and 661 mA. Here, 3D virtual models, representative of various sections of membranes, were built, and the data were mathematically managed by CT Analyzer software, v. 1.13.5.2.2.8. Statistical Analyses Differences among nisin concentrations are presented as mean ± standard deviation. ANOVA (analysis of variance) was used for parametric data, followed by Duncan or Scheffe tests for multiple comparisons. A p value < 0.05 was accepted as statistically significant. The data were analyzed using Statistica ® 8.0 (Statsoft software, Tulsa, OK, USA). Nisin Standard Curve For the nisin standard curve, zones of inhibition showed different sizes of halo ranging from 14 to 36.30 mm. Each halo corresponded to one concentration of nisin ranging from 2.5 g/mL to 2500 g/mL, as seen in Figure 1. The standard curve demonstrates the sensibility of the bioindicator L. sakei even when it was exposed to low nisin concentration. This finding is important in establishing the relationship between the amount of nisin and the method that was used. Nisin is a soluble peptide and its low molecular weight facilitates its diffusion in other matrices, supplementing the antimicrobial activity. The standard curve demonstrates the sensibility of the bioindicator L. sakei even when it was exposed to low nisin concentration. This finding is important in establishing the relationship between the amount of nisin and the method that was used. Nisin is a soluble peptide and its low molecular weight facilitates its diffusion in other matrices, supplementing the antimicrobial activity. Evaluation of Nisin Loading in BC by Time The nisin concentration was set at 250 g/mL and we were able to observe the loading efficiency (%LE) by amount of protein (mg/mL) and nisin activity (log 10 AU) at different periods ( Table 2). The protein concentration was not directly proportional to AU. Despite different incubation periods, there was little variation in nisin titers as observed in Table 2. These results are key to the definition of the best time for the load efficiency. After 4 h, the N-BC with 45% proteins was almost 2 log 10 higher than the initial solution. Once the nisin solution was not pure, it is possible to suggest that the BC may have acted as a filter, separating the nisin from the other components and maybe causing overly strong adsorption kinetics in 4 h. The same observation was reported by Ataide and colleagues ; they evaluated the enzyme bromelain loaded into the BC membranes, and its release. They noticed a threefold increase in the specific bromelain activity when compared to the initial solution, with 31% of proteins when bromelain was loaded into BC, showing the selective behavior of the BC membrane. Jorge et al. studied a system based on BC for the stabilization of insulin. The results indicate that BC was able to enhance permeation. Nguyen et al. also evaluated the nisin loaded in the BC membranes. However, they reported 6 h as the best period for nisin incorporation in the BC membranes, two hours longer than in our study. Moreover, Nguyen and colleagues used 600 IU/mL in the initial solution, and we experimented with 10,000 AU/mL (4 log 10 AU). According to Moniri et al., BC has been extensively studied for controlled drug delivery as its structure is suitable for the dispensing of biomolecules. This structure, which consists of nanofibers, as reported in the SEM section, probably explains the enhancement of nisin's activity when loaded into BC. The BC nanofibers could act as a filter, retaining big molecules of dirt from the nisin solution and selectively allowing the absorption of the low-weight molecules, such as nisin. Figure 2A shows the qualitative agar diffusion test of N-BC against L. sakei (Figure 2(Aa)) and S. aureus (Figure 2(Ab)) after 7 days of storage at room temperature. Additionally, the quantitative tests of stability against L. sakei and S. aureus are shown in Figure 2B,C, respectively. Figure 2A shows the qualitative agar diffusion test of N-BC against L. sakei ( Figure 2(Aa)) and S. aureus (Figure 2(Ab)) after 7 days of storage at room temperature. Additionally, the quantitative tests of stability against L. sakei and S. aureus are shown in Figure 2B,C, respectively. In general, the N-BC showed antimicrobial activity against S. aureus for 60 days at 4 and 25 C. No antimicrobial activity was observed for 60 days at 37 C. Despite having antimicrobial activity, N-BC was statistically stable for up to 7 days at 4 and 25 C. At 37 C, N-BC was stable for up to 3 days. After these periods of time, a significant decrease in activity was revealed. Stability Test by Agar Diffusion Assay On the other hand, N-BC was stable for up to 100 days against L. sakei at all incubation temperatures ( Figure 2C), and no significant difference was seen. By observing these results, we could hypothesize that if the N-BC system was used as wound dressing in tissues contaminated with S. aureus, it should be replaced-or even reapplied-after 7 days. Furthermore, if the N-BC system was used in food packaging, it should be considered for fresh food wrapping. The process of nisin purification, described in the literature, shows that the activity of nisin can be reduced by fat clusters, salt concentrations and the aggregation of soluble proteins. It also shows that, after purification, nisin's antimicrobial activity was increased. Staphylococci are bacteria that live on the epithelial surfaces of humans and animals, being responsible for food poisoning and infections. With respect to bacterial resistance, nisin has been proven to be useful for limiting the development of antimicrobial resistance. S. aureus also appears among the main bacteria responsible for secondary infections in burns. Nisin's antimicrobial activity against S. aureus generated a significant decrease in viable cells in a period of 1 day, and even inhibited biofilm formation. Nisin encapsulation in nanofiber polymer provides stability to nisin for a period of up to seven days. However, longer periods produced an inhibition zone inferior to what is presented in our study. Figure 3 shows the cytotoxicity data of N-BC at 24 and 48 h, at different concentrations. The fibroblast cells did not exhibit any significant change after 24 h of treatment. On the other hand, after 48 h, proliferation rates were 25% higher than those produced after 24 h, indicating the N-BC was able to stimulate cell proliferation, and no toxicity was reported. Figure 3 shows the cytotoxicity data of N-BC at 24 and 48 h, at different concentrations. The fibroblast cells did not exhibit any significant change after 24 h of treatment. On the other hand, after 48 h, proliferation rates were 25% higher than those produced after 24 h, indicating the N-BC was able to stimulate cell proliferation, and no toxicity was reported. Nisin is a safe molecule for both FDA and WHO, and BC is also considered a non-cytotoxic material. Corroborating the Gao et al. study, a cytotoxicity test of N-BC was performed in a co-culture and no cytotoxicity was observed. Proliferation in fibroblasts is important to accelerate the healing process. Cytotoxicity Assay The BC membranes loaded with nisin were used in large animals to treat surgical dehorn wounds. The authors observed that the treatment with the BC membrane accelerated the healing process. Wound healing involves a dynamic set of tissue changes, important for maintaining the integrity of an organism. The authors also noted that BC may have created a barrier between the wound and the environment, preventing contamination. In a previous work, dos Santos et al. analyzed the antioxidant activity of nisin. The antioxidant activity may be related to the proliferation of fibroblasts since nisin has the potential to neutralize free radicals, facilitating cell proliferation. Nisin is a safe molecule for both FDA and WHO, and BC is also considered a non-cytotoxic material. Corroborating the Gao et al. study, a cytotoxicity test of N-BC was performed in a co-culture and no cytotoxicity was observed. Proliferation in fibroblasts is important to accelerate the healing process. The BC membranes loaded with nisin were used in large animals to treat surgical dehorn wounds. The authors observed that the treatment with the BC membrane accelerated the healing process. Wound healing involves a dynamic set of tissue changes, important for maintaining the integrity of an organism. The authors also noted that BC may have created a barrier between the wound and the environment, preventing contamination. In a previous work, dos Santos et al. analyzed the antioxidant activity of nisin. The antioxidant activity may be related to the proliferation of fibroblasts since nisin has the potential to neutralize free radicals, facilitating cell proliferation. Figure 4A,B shows the BC without and with nisin, respectively. Comparing the morphology, it is possible to identify a similarity between the fibers on a micrometric and nanometric scale for both forms of BC. However, in N-BC, the fibers were more swollen than in BC, suggesting nisin incorporation. Figure 4A,B shows the BC without and with nisin, respectively. Comparing the morphology, it is possible to identify a similarity between the fibers on a micrometric and nanometric scale for both forms of BC. However, in N-BC, the fibers were more swollen than in BC, suggesting nisin incorporation. The fiber network accounts for a large surface area and the porous structure of the BC facilitates the immobilization of nisin. In addition, this structure allows the diffusion of water, providing a functional stabilization of the biomolecules. X-ray Microtomography The quantitative results of the structure were confirmed by X-ray microtomography The fiber network accounts for a large surface area and the porous structure of the BC facilitates the immobilization of nisin. In addition, this structure allows the diffusion of water, providing a functional stabilization of the biomolecules. X-ray Microtomography The quantitative results of the structure were confirmed by X-ray microtomography assay, as presented in Table 3. The BC membranes presented more connectivity than the N-BC ones. Additionally, the presence of nisin decreased the porosity of membranes by 2%, supporting the SEM images. The pores were interconnected and distributed throughout the sample. The BC structure had a lower degree of anisotropy than the N-BC membrane. Table 3. X-ray microtomography parameters of the bacterial cellulose (BC) and the bacterial cellulose with nisin (N-BC) membranes. Regarding membrane morphology, Gedarawatte et al. performed the SEM test in nanocrystal BC containing nisin, and the presence of the antimicrobial was identified in the swelling of samples. The same behavior was observed in BC and nisin samples examined in co-culture by Gao et al., reporting a large presence of nisin in the morphology of swelling fibers. The increase in fiber size indicated a high amount of antibacterial substance loaded in the BC membrane. Gao et al. additionally observed that nisin can affect the BC fibers, resulting in a slight decrease in crystallinity and improving the antimicrobial property. Conclusions Here, we have proven the possibility of loading nisin into the BC membranes in 4 h, and also improving the antimicrobial activity with less protein, which suggests the BC membrane served as a selective barrier and it separated nisin from other components. This barrier enhanced nisin's activity, and as a consequence of the nisin loading, a stable N-BC system was created and considered ideal for nisin delivery, with no toxicity and capacity to control the growth of microorganisms. Thus, N-BC has the potential to be used as a dressing for skin wounds since nisin may prevent infections upon its release, requiring only a replacement at a point between 3 and 7 days. Moreover, N-BC can be applied in in food packaging and other fields because its antimicrobial activity will probably reduce food deterioration and improve shelf-life.

Scrutiny of the veracity of Ben Carson’s account of various events in his youth continues, with the Wall Street Journal casting doubt on Carson’s claim to have sheltered white students in his high school biology lab during riots following the 1968 assassination of Martin Luther King, Jr., and on his claim about events in a psychology class at Yale. Some conservatives (including Carson himself) lashed out at Politico yesterday over its reporting on Carson’s claim to have been offered a full scholarship to West Point—a headline in the Federalist even referred to the essential read for Beltway insiders as a “leftist rag.” (Someone, please, send the Federalist a subscription to the Socialist Worker.) But today some reliably conservative outlets seem more chastened. “WSJ: More Ben Carson Narratives Can’t Be Confirmed,” reads the sober Newsmax headline. “More stories from Ben Carson’s past that no one can authenticate,” Hot Air reports dispassionately. As Paul Waldman writes in an astute piece at the Washington Post, this scrutiny of the accuracy of Carson’s autobiography tells us little about what kind of president he’d be. But Carson’s wild claims, say, about the purpose of the pyramids, the origins of the universe, or his rejection of evolution, Waldman contends, “suggest not only that his beliefs are impervious to evidence but also an alarming lack of what we might call epistemological modesty.” “Some parts of his personal story are irrelevant to that assessment” of his suitability for the presidency, Waldman concludes, “but some parts aren’t. And it’s those that should really give us pause.” But there’s a way to see Carson’s apparent dissembling on a variety of issues, from his personal story to his views on science and history, as of a piece. That piece consists of a very simple frame: that Carson, through his faith in God, the grit exemplified in his life story, and the smarts evidenced by his success as a neurosurgeon, sees the truth and, as a presidential candidate, is conveying that truth to the American people. Questioning his claims about history and the universe’s origin are akin to questioning Carson’s own origin story. Ben Carson believes he has seen the enemy, and casts himself as a prophet warning America of it. Carson’s method of disarming his critics is to portray them as that enemy in a cosmic battle over that truth. The enemy is people Carson facilely refers to as “secular progressives.” Secular progressives, to Carson, are not mere political adversaries. In Carson’s usage, the term seems to encompass any sort of person or entity that might fall into categories as varied as liberal, secular, religious liberal, religious progressive, communist, socialist, feminist, LGBT rights activist, civil liberties advocate, Someone Who Disagrees With Ben Carson, atheist, agnostic, dormant and dead moderate Republicans, and, oh, I don’t know, CNN? Yesterday, as the Huffington Post’s media editor Gabriel Arana reports, a Carson interview on CNN turned “strangely combative” as anchor Alisyn Camerota “pressed him on a number of his recent controversial comments, including the claim that ‘many’ Americans are stupid and that ‘we’d be Cuba if it weren’t for Fox News.‘” Camerota reminds Carson she used to work at Fox News and still has many friends there, but this appears to be of no moment to Carson, who lectures her (emphasis mine): the general mainstream media all seems to move in the secular progressive direction, and you know, they would like to create a narrative that certain things are good, and certain things are bad, according to the way that they see them. And by being able to be the bully pulpit, so to speak, and to be the only voice that’s out there, you can get a lot of people to start thinking the way that you do. Along comes Fox News and presents an alternative, a different way of thinking. Not only do “secular progressives” try to discredit Dr. Ben Carson; he maintains they are trying to silence him and maybe even harm him. Last month, Carson claimed he needs Secret Service protection because “I’m in great danger because I challenge the secular progressive movement to the very core.” Carson also appears to see himself as a prophet warning of the subterfuge used by “secular progressives.” As is evident if you watch the exchange with Camerota, or Carson’s hectoring news conference yesterday, questioning him is trying to “change the subject.” For Carson, “changing the subject” is “secular progressives'” way of silencing him. As he wrote in his 2014 book, One Nation: What We Can Do To Save America’s Future (emphasis mine): It is particularly important when dealing with adversaries to know what points you want to make while remaining focused. This makes constant interruptions, attacks, and attempts to change the subject more difficult. If you are an effective representative of American values, the secular progressives will make every attempt to destroy your character by exposing any mistakes, misstatements, or misdeeds from you past. Naturally, there are no perfect people, present company included, which makes the threats of exposure extremely potent. In the book, Carson offers examples of these alleged secular progressive efforts to destroy not only his character but the country. He claims, broadly, that “secular progressives have zoomed past the intent of the [First Amendment] and tried to replace it with their anti-God propaganda reinforced by bullying tactics.” They have attacked him in particular, arguing, for example, that “secular progressives seized upon the opportunity to distort what I said” about same-sex marriage. In Carson’s mind, though, he’s not a victim but an embattled hero. “I have been spreading the word that we must have enough backbone to stand up to secular progressives who insist on fundamentally changing America into something we would not recognize,” he wrote in One Nation. He keeps speaking out, he continued, “despite the many efforts of secular progressives to discredit and silence me.” What’s funny (not really) about Carson’s litany of alleged “secular progressive” sins is that he seems oblivious to his own failures to live up to the very demands he makes of others. In yesterday’s news conference, for example, he was asked about President Obama’s decision to reject the Keystone XL oil pipeline. Carson, who believes in creationism and rejects evolution, criticized Obama for basing his decision on “ideology” and “not on evidence.” While he boasts of being a protector of the Constitution and the First Amendment in particular, Carson questioned the assembled journalists whether they would “sing my praises” and promise “none of this stuff will ever go on again” if he would reveal the name of the person he allegedly tried to stab as a teenager—another story from his autobiography that lacks corroboration. A free press, apparently, is just another secular progressive tool. Carson seems certain that using the “secular progressive” straw man to insulate himself from criticism will work with voters. “All you guys trying to pile on is actually going to help me,” he predicted confidently at yesterday’s news conference. It may work with a Republican base conditioned to distrust the media as shills of the left and enemies of religion, the same people who might believe, as Carson does, that America would be like Cuba if it weren’t for Fox News. As Ed Kilgore and David Corn have documented, many of Carson’s beliefs have long roots in the conspiratorial American right dating back to the Cold War, so he’s tapping into a deep well. But as Heather Parton has repeatedly pointed out, Carson’s method of attacking his perceived enemies (even the National Review!) undermines his reputation as a soft-spoken, reliably nice guy, the crucial underpinning of his candidacy. Since he’s so—shall we say—liberal with his use of the “secular progressive” indictment, the question now is whether he accuses so many enemies that he runs out of targets.

The major broadcast networks failed to persuade an appeals court to shut down Aereo Inc., the Barry Diller-backed online TV service that they claim violates their copyrights. Watching the NFL online using Aereo's service Credit: Aereo Broadcasters including Walt Disney Co.'s ABC and Comcast's NBC unsuccessfully petitioned the U.S. Court of Appeals in New York to overturn a lower-court order denying a preliminary injunction that would have put New York-based Aereo out of business. "Plaintiffs have not demonstrated that they are likely to prevail on the merits on this claim in their copyright infringement action," the appellate judges said in the opinion. The networks sued Aereo in March 2012, claiming that it infringed copyrights by capturing their over-the-air signals and retransmitting the programming to subscribers on computers and smartphones without paying for the rights. The networks argued that Aereo would devalue their programming and cut viewership, jeopardizing revenue from advertisers and pay-TV providers. "Aereo is a retransmission service by its own design," Bruce Keller, a lawyer for the broadcasters, told the three-judge appeals panel during oral arguments in November. The networks said that Aereo, by streaming programs to its subscribers, engaged in public performances of their copyrighted works, which requires a license. U.S. District Judge Alison Nathan had denied an injunction in July, ruling that Aereo's retransmissions weren't public performances. Aereo said that its service gives subscribers access to broadcast programming and lets them record it using remotely located individual antennas and digital video recorders for playback later. That constitutes a private performance under copyright law, Aereo said. "Customers have the right to make private performances," David Hosp, a lawyer for Aereo, told the appeals court in November. "Supplying the technology to accomplish this does not violate the public-performance right." One analyst said the ruling may not be as significant as it seems, saying the litigation process has a long road ahead before arriving at a final answer. "Further, the scope of the argument for the pre-trial injunction was somewhat narrow and our Industry sources say the broader scope in the full trial makes the pre-trial injunction ruling potentially less relevant than it appears," said the analyst, David Bank of RBC Capital Markets, in a research note following the ruling. Despite all the "noise" around Aereo, the broadcasters' business will ultimately absorb "any outcome without major disruption," Mr. Bank added. ~ Bloomberg News and Ad Age staff ~

A typical connector assembly includes a male connector generally in the form of a plug and a female connector generally in the form of a ferrule which forms a socket. In use, the male connector is plugged into the female connector to effect a mechanical and electrical connection between the two. Typically, a cable in the form of a coaxial cable is electrically and mechanically attached to one of the connectors such as the male connector, and the other connector, such as the female connector, is electrically and mechanically attached to another coaxial cable or to a circuit such as a circuit on a printed circuit board. An example of such a connector assembly is a typical antenna connector assembly for antenna cables such as those used in the automobile industry for radios. Such a connector assembly typically includes a male connector and a female connector which are mechanically and electrically attached to coaxial cables and in use are mechanically and electrically connected to each other. In such prior art devices, the lack of satisfactory tactile feedback makes it difficult to know when a suitable connection has been made. In addition, if the connectors are not locked together there may be a tendency for the connectors to be inadvertently disengaged. Connector latches known in the art for use in locking together male and female connectors tend to be bulky and tend to require unusual effort to separate once engaged. Such connector latches also tend to be expensive.

Human papilloma virus in adolescence. Human papilloma virus (HPV) is one of the most widespread sexually transmitted diseases especially in adolescence. The majority of the infections are self-sustained by the immune system. However, HPV may lead to genital warts, cervical dysplasia and cervical cancer. Sexually active adolescent females should be encouraged to obtain gynecologic screening for HPV and be well informed about HPV and the risks associated with this infection. All the efforts are now focused on the vaccines that are being developed to reduce the morbidity and mortality associated with HPV infection if administered in time.

It took around fourteen San Francisco cops to strong-arm and subdue a homeless Black man after they accused him of being armed with his crutches. To make matters worse, they not only forced his prosthetic leg loose but also refused to pull his pants up after he had been flattened to the ground. It took 14 San Francisco Police Department officers to take down and restrain a one-legged, black homeless man, armed with crutches and apparently dangerous. The incident is the latest embarrassment for the US police, who face constant accusations of unreasonable use of force. The confrontation was captured on video released by journalist Chaedria LaBouvier via blog platform Medium, and shows white police officers taking down a one-legged homeless black man on the city’s central Market street. According to witnesses, police were called in to the scene to take care of a suspicious man waving some “sticks” around. The video of the incident which happened on August 4, shows the extent of humiliation and brute force exercised immediately after the man was wrestled to the ground by SFPD officers. As the disabled male struggles to move, cops pin him down. “These are my crutches. I use these to walk,” the man tried to explain. But even after realizing that the man had a prosthetic leg, the police continued to use overwhelming physical restrain and man-handled him, forcing his head to the ground. Beaten to the ground, the suspect at one point said, “what the f**k is you doing this to me?” as more officers arrived to form a cordon around the incident area. “Is this respectable? When I say ‘no’, is this what you do to me?” the man said. Witnesses spoke out against police brutality from the start of the video but to no avail as cops continued to abuse the one legged man. The camera operator especially noted the “lack of respect” for the suspect as clothes were pulled off the man during the incident. First one can see the man’s buttocks being exposed, and minutes later his entire back. “But don’t let it be a black and a white one/Cause they’ll slam ya down to the street top/Black police showing out for the white cop…” — Ice Cube. Watch the San Francisco cops do their best attempt at “policing” in the video above.

. Comparison was made of the effect of total adrenalectomy on the gastric secretion in chronic experiments on dogs with the Pavlov's stomach and Basov's fistula. A decrease of the maximal secretion level of gastric juice was associated with the alteration of the organ hemodynamics. A tendency to reduction of the acid production in the stomach was revealed. Essential differences were noted in the character of proteolytic enzymes secretion with different agents stimulating the secretion. Specific nature of the gastric secretory system for each stimulant, and different effects of adrenalectomy on the secretion induced by these stimulants was shown.

Outcome of Endovascular Treatment in Symptomatic Intracranial Vascular Stenosis Objective The outcome evaluation for the revascularization of intracranial vascular stenoses has not been fully described due to the highly technical nature of the procedure. We report here on the early and late clinical outcomes of angioplasty and/or stenting of symptomatic severe intracranial vascular stenoses at a single institute. Materials and Methods Since 1995, we have treated 35 patients with symptomatic intracranial vascular stenosis (more than 70% stenosis, mean stenosis: 78.6% ± 6.2%). Angioplasty (n = 19) was performed for the horizontal segment of the middle cerebral artery (M1) (n = 16) and the basilar artery (BA) (n = 1), the intradural vertebral artery (VA) (n = 1), and the cavernous internal carotid artery (ICA) (n = 1). Stenting (n = 16) was performed for the cavernous or petrous ICAs (n = 9), the intradural VA (n = 3), BA (n = 2), and M1 (n = 2) artery. We assessed the angiographic success (defined as residual stenosis < 50%) rate, the periprocedural complications during the 30-day periprocedural period, the symptomatic recurrence and restenosis during a mean 22-month follow-up (FU) period. The Kaplan-Meier estimate of the cumulative event-free rate of the major cerebrovascular events, i.e. death, stroke or restenosis, was also done. Results Angiographic success was achieved in 97% of our patients (34/35). There were four procedure-related complications (11%) including a death and a minor stroke. During the mean 22-month FU, the asymptomatic restenosis rate was 9% and the symptomatic restenosis rate was 6% in the target lesion and 9% in all the vascular territories. The Kaplan-Meier estimate was 70.6% (95% confidence interval = 46.5-94.7) after 33 months of FU. Conclusion In addition to a high angiographic success rate and an acceptable periprocedural complication rate, intracranial angioplasty and/or stenting revealed a relatively low symptomatic recurrence rate. Hemorrhage is a rare, but the physician must aware that potentially fatal periprocedural complications can occur. ntracranial atherosclerosis is a major cause of ischemic stroke, and depending on the studied population, it accounts for 8 15% of all strokes that are due to cerebral atherosclerosis (1 5). In a retrospective, nonrandomized review of 151 patients in the United States, the annual rate of stroke for patients with symptomatic severe intracranial stenosis was 10.7% in the aspirintreated group. Annual stroke rates reach 15.0% for patients with basilar artery stenosis and 13.7% for patients with vertebral artery stenosis. Thijs et al. have reported that those patients with symptomatic intracranial atherosclerosis who failed at antithrombotic therapy, they revealed a very high rate of cerebral ischemic events (55.8%), e.g., 29 of the 52 study patients. I Extracranial-intracranial anastomosis has not been proven to be effective for preventing cerebral ischemia in those patients with atherosclerotic arterial disease in the carotid and middle cerebral arteries. In the report by the EC/IC Bypass Study Group, non-fatal and fatal strokes occurred earlier and more often in patients who underwent bypass surgery and the patients with severe middle cerebral artery stenosis had the worst outcomes. Bypass surgery also resulted in an 8% mortality rate and a 13% morbidity rate for 83 patients having symptomatic vertebrobasilar insufficiency who underwent 85 bypass procedures. Although endovascular revascularization for symptomatic intracranial stenoses remains at the investigational stage and much of the pertinent information is anecdotal, intracranial angioplasty and stenting are being increasingly performed to treat stenotic lesions. The complication rates range from 0 50% (12 23). The possible reasons why the outcomes of angioplasty and stenting procedures vary greatly from center to center have not yet been identified. In addition, any direct comparison of the relevant literature results is difficult because there is no uniformity of the procedures' protocols and of the methods for evaluating the results, and there are also ethnic differences in the incidence of intracranial stenosis. For Koreans, severe atherosclerotic stenoses tend to be more intracranial stenoses than extracranial stenoses. We report here on our results for angioplasty/stenting of the intracranial arteries in 35 consecutive patients. We have included the early and late clinical outcomes, and we compared our results to the results that have been reported in the literature. MATERIALS AND METHODS Since 1995, we have treated 35 consecutive patients with severe (more than 70%) symptomatic intracranial stenosis. Nine patients from the authors' previous reports were included along with their full follow-up (FU) data. The patients' ages ranged from 31 to 73 years (mean age: 53), and the male to female ratio was 3.4:1. Angioplasty (n = 19) was done in the M1 segment of the middle cerebral artery (M1) (n = 16) (Figs. 1, 2), the basilar artery (BA) (n = 1), the intradural vertebral artery (VA) (n = 1), and the cavernous internal carotid artery (ICA) (n = 1). Stenting (n = 16) was done in the cavernous (n = 8) or petrous (n = 1) ICA (Fig. 3), the intradural VA (n = 3), the basiler artery (BA) (n = 2), and the M1 segment of the middle cerebral artery (M1) (n = 2). This report did not include any patients who were treated as part of emergency protocols or those patients with intracranial occlusion. For each patient, a complete history taking and a neurological examination were performed by an independent neurologist who was not involved in the interventional procedure. Our data revealed that types of procedure were different among the lesion sites; angioplasty was performed more often in the M1 segment in contrast to stenting being performed in the posterior circulation and in the cavernous segment of the ICA (p < 0.001). Angio-interventional Procedures Most patients were premedicated with 100 mg of acetylsalicylic acid (ASA) and either 75 mg of clopidogrel (Plavix, Sanofi, Inc) or 250 mg of ticlopidine (Ticlid, Roche, Inc) for at least four days before the procedure. One hundred milligrams of ASA (low dose regimen) once daily was continued as a permanent medication. Additionally, 75 mg of clopidogrel were given once daily for more than six months following the procedures. All the procedures were performed under local anesthesia, and during the procedures, each patient received 5,000 8,000 IU of intravenous heparin to attain an activated clotting time (ACT) of more than 250 seconds. Prior to treatment, diagnostic cerebral angiography was performed via the transfemoral approach. The procedure started with a common carotid artery injection, and this was followed by selective angiography of both the internal carotid and One-year follow-up angiogram shows good patency of the stented segment of the internal carotid artery with a 20% stenosis at the distal stent margin. Note a patent side branch of the cavernous segment of the internal carotid artery. The patient recovered completely and had no symptom recurrence during the one-year follow-up period. A B C vertebral arteries. A 6F sheath was introduced and it was then positioned in either the internal carotid artery or the vertebral artery. The sidearm of the guiding catheter was continuously flushed with pressurized, heparinized normal saline, and this catheter was used for angiography treatment steps. After crossing the lesion with a 0.010 0.014-inch outer diameter microguidewire, the balloon catheter was placed over the microguidewire and next directed across the lesion. We used balloon catheters with diameters ranging from 2 to 3 mm and with lengths of 10 or 20 mm according to the target vessel size, and we were careful not to allow any over-dilatation. Balloon inflation was performed slowly and maintained for 10 to 20 seconds with a disposable inflation device that allowed for exact pressure control. In the 16 stenting procedures, the stenotic lesion was predilated with a balloon catheter 2 mm in diameter and 10 20 mm in length in all cases. A stent 2 4 mm in diameter and 8 18 mm in length was passed over the guidewire and positioned across the stenosis. Each stent was selected according to the size of the target vessel. The residual stenosis of the treated intracranial artery was measured via post-procedural angiography. As the more flexible and lower profile stents have become available, our approach has changed from performing angioplasty alone to performing angioplasty followed by stenting at the same session: this has been done on a more routine basis than was previously done before 2002. Measurement of Stenosis The percentage of the diameter of the stenosis was calculated by dividing the narrowest linear diameter at the stenotic segment by the distal diameter at the segment of the same vessel that had a normal appearance. Lesion length and eccentricity were measured and categorized according to Mori's classification and these parameters were analyzed for comparison of treatment outcome. An eccentric lesion was defined as when the residual lumen was asymmetrically shifted to one wall of the vessel due to the eccentrically located plaque. Restenosis was evaluated by using transcranial Doppler and/or cerebral angiography. FU Evaluation We obtained the clinical data, with the exception of the data for one patient who died, during a mean 22-month FU period (FU range: 3 86 months, median FU: 13). In addition to the clinical FU, MR angiography (MRA) was obtained in 11 patients (Figs. 1, 2), conventional angiography was done for 10 patients (Fig. 3), CT angiography was done for one patient, and transcranial Doppler (TCD) was done for only five patients. The mean imaging study FU period including the TCD was 21 months (range: 3 74, median: 11). Only the clinical FU without the imaging studies was available for seven patients. Two of these seven patients were followed for less than six months (three and four months) and other five patients did not undergo angiographic FU. Evaluation of Outcome and Statistical Methods We evaluated angiographic success rate, the periprocedural complications during the 30 days periprocedural period, and the symptomatic recurrence and restenosis during a mean 22-month FU period. The angiographic success, as calculated on a per-vessel basis, was defined as any residual stenosis < 50%. Angiographic restenosis on conventional cerebral angiography was defined as a stenosis > 50% of the luminal diameter. MR angiography FU was mainly used for those patients who only received angioplasty and the degree of stenosis on MRA was based on the interpretations by the staff neuroradiologists. Doppler ultrasonic restenosis was defined as an increased peak systolic velocity threshold of 220 cm/sec for the intracranial internal carotid artery and the proximal middle cerebral artery; the corresponding thresholds were 140 cm/sec for the basilar artery and 120 cm/sec for the vertebral artery. Minor stroke was defined as a new non-disabling neurological deficit or an increase of 3 on the National Institutes of Health Stroke Scale, or a nonfatal hemorrhage that completely resolved within 30 days. Major stroke was defined as a new neurologic deficit that persisted beyond 30 days and an increase of 4 according to the National Institutes of Health Stroke Scale. The cumulative event-free rate of major adverse cerebrovascular events (MACEs), i.e. death, stroke or restenosis was analyzed by the Kaplan-Meier method. MACEs included stroke, death and restenosis with or without a symptomatic recurrence during the FU. The logrank test was used to compare MACEs between the M1 segment of middle cerebral artery and the other sites in the intracranial arteries. We used x 2 or Fisher's exact tests to compare the differences of MACEs between angioplasty and stenting procedures and for the lesion length, lesion eccentricity and the location of the lesions. All the calculations were performed with SPSS for Windows (version 10.0, SPSS Inc). Repeated angioplasty or repeated stenting during the FU period was not attempted for our patients. We compared our results with the relevant long-term outcomes in the currently available literature (Table 1). RESULTS The mean degree of stenosis in the symptomatic intracranial artery was 78.6% 6.2%. After the procedure, the mean residual stenosis was 20.7% 14.4%. Angiographic success (less than 50% residual stenosis) was achieved in 97% of the patients, and the only exception was one patient who had an asymptomatic dissection (34/35). Periprocedural complications within the first 30 days were noted in four patients (11%), and this included an aymptomatic dissection at M1 and a transient ischemic attack at M1. There was a minor stroke in one patient (3%) who experienced a subarachnoid hemorrhage that was due to a guidewire-related arterial perforation, but it resulted in only a transient headache. There was one death due to hemorrhage during a BA stenting (3%). There was no recurrence of symptoms within one year period (Figs. 1, 3). During the mean 22-month total FU period, symptomatic restenosis/occlusion related to the target lesion was noted in two patients (symptomatic restenosis, 6%). One patient had an ischemic stroke that was not related to the treated vessel. Asymptomatic restenosis was detected in three patients (9%) during the FU period. Among the three symptomatic patients we observed during FU, one patient experienced aggravation of a neurologic deficit that was related to an occlusion at the M1 segment 24 months after the angioplasty procedure (Fig. 2). Another patient had an intracerebral hematoma and intraventricular hemorrhage that was associated with M1 occlusion 16 months after the angioplasty procedure, but this patient experienced a full recovery by the time of the 33-month FU examination. The other patient revealed an ipsilateral internal carotid artery occlusion on MRA at the time of a pontine infarction 20 months after the procedure; this led to some mild weakness of the patient's extremities. The Kaplan-Meier curves predicted that the cumulative event-free rate of MACEs, i.e. death, stroke or restenosis, was 70.6% (95% confidence interval = 46.5 94.7) at 33 months. The log rank test for the difference in the cumulative event-free rate of MACEs according to the lesion site showed no statistical significance. There were also no significant differences in MACEs among the types of procedures, the lesion lengths and the lesion eccentricities. DISCUSSION Our experience in this study indicated that 97% of our study patients achieved angiographic success (34/35). The incidence of symptomatic restenosis was very low after angioplasty and/or stenting of intracranial stenoses, i.e. 6%, during a mean 22 months of FU. The symptomatic recurrence rate in any vascular territory was 9% during the same FU period. However, intracranial angioplasty and stenting had a 3% fatal complication rate, although our fatality rate was less than those fatality rates reported in the literature (Table 1). Because the symptomatic recurrence rate was as high as 9% and it seemed to go higher with further long term FU, the physician is required to make a therapeutic decision for those patients with asymptomatic restenosis. Our policy has been to observe these patients closely while they are under medication because symptomatic restenosis is regarded as the only indication for a revascularization procedure including repeated angioplasty and/or stenting. The recent advent of new generation stents that have low profiles and high flexibility has encouraged the consideration of performing stent-assisted angioplasty as an alternative approach to angioplasty alone for intracranial stenosis (17 23). Presently, the primary limitation of angioplasty appears to be restenosis (defined as more than 50% diameter stenosis on FU angiography), and this is seen in as many as 30% to 40% of the patients who are treated by coronary angioplasty. The complication rates of intracranial angioplasty and stenting are known to be very high, i.e. from 0 to 50% (12 23). The mortality rate was 3% in our study. The periprocedural mortality and morbidity rate also seem to be higher after stenting than for an angioplasty procedure alone. Bleeding is a serious complication for intracranial stenting as was seen in one of our study patients. The causes of bleeding include a bleeding tendency caused by aggressive antiplatelet and anticoagulation therapy, hyperperfusion syndrome, vessel rupture or dissection due to ballooning and/or stenting, and guidewire injury. The subarachnoid hemorrhage due to a guidewire injury in a patient of our study had a relatively benign clinical course without further symptomatic problems at four months clinical FU. In addition to the hemorrhagic complications, infarctions related to thromboemboli or to side-branch occlusion due to the presence of the deployed stent also contribute to the fatal peri-procedural complications. We only experienced one transient ischenic attack in a patient after angioplasty of the M1 segment. One of the major concerns in intracranial stenting is the side branch occlusion that can be caused by the stent struts, although we did not experience any problem related to the perforator or side branch occlusion. The possible mechanism for side branch occlusion after stenting in the coronary artery is the 'snow plow' effect where the atheroma from the parent vessels is shifted into the ostium of a side branch. Other mechanisms for side branch occlusion may include side branch spasm, embolization of the atherosclerotic material, thrombus formation and the stent material itself. The presence of ostial narrowing arising from within or just beyond the diseased portion of the parent vessel was the most powerful predictor of side branch occlusion in the coronary artery immediately after stenting. The plaque volume of the parent vessel and the side branches is a major determinant for the fate of the side branches. Perforaters in M1 and BA are different from the side branches in the coronary vessels in that the perforators are terminal branches supplying the deep nuclei of the basal ganglia and brainstem. Experimental evidence from canine vertebral arteries suggests that the cervical muscular branches, which are comparable to human perforators, tend to remain patent if less than 50% of the ostial diameter is covered by the stent strut. A size comparison between the stents and the perforators should be made in order to avoid any compromised flow in the perforators. In conclusion angioplasty and/or stenting of the intracranial arteries are relatively safe procedures having a high angiographic success rate (97%), an acceptable rate of periprocedural complication (11%), which included a minor stroke and a death within 30 days, and a low symptomatic restenosis rate for the target lesion (6%) and for all the vascular territories (9%) during the mean 22month FU. Replacement of intracranial revascularization by stenting needs to be further studied in conjunction with the advent and arrival of new stent devices that have high flexibility and low profiles.

News items related to Windows 7 migration as issued by the Send2Press Newswire service on behalf of the noted news source. NEW YORK, N.Y. (SEND2PRESS NEWSWIRE) — At the end of the year, through year-long use for work and pleasure, your hard drive has been stuffed with all kinds of materials, both business and personal. Perhaps you are planning a comprehensive system partition optimization or hard drive management task, such as extending the fully crammed system partition for smooth and fast Windows performance, or resizing and moving partitions or partitioning the newly upgraded hard disk. NEW YORK, N.Y. (SEND2PRESS NEWSWIRE) — EASEUS, the security and storage management software producer, releases the newly updated Windows 7 compatible system backup and disaster recovery freeware: EASEUS Todo Backup 1.1. The new version is completely upgraded for Windows 7 system and hard drive backup. It can be used to back up system state, applications, and settings as well as the hard disk partition.

A Review of Risk Management Techniques Growth of the Nigeria Economy Risk management is a subject of interest in the contemporary Nigerian investment climate; most especially when the world is currently facing several devastating dangers or risks of extinction or collapse of national economies through dreaded HIV/AIDS infections, stress or high blood pressure (hypertension) leading to stroke attacks; due to uncertainty and job insecurity in the economy. Besides, because of rampant frauds and corrupt practices in our society today, people have virtually lost trust and confidence in thebusiness and other social relationships. Moreover, there is an ever growing trend of unemployment rate and incessant armed robbery cases, insurgency by the Boko Haram terrorists in the North East collapse of buildings due to faulty construction specifications which have compounded mans living standard in Nigeria. Introduction Risk management is a subject of interest in the contemporary Nigerian investment climate; most especially when the world is currently facing several devastating dangers or risks of extinction or collapse of national economies through dreaded HIV/AIDS infections, stress or high blood pressure (hypertension) leading to stroke attacks; due to uncertainty and job insecurity in the economy. Besides, because of rampant frauds and corrupt practices in our society today, people have virtually lost trust and confidence in the-business and other social relationships. Moreover, there is an ever growing trend of unemployment rate and incessant armed robbery cases, insurgency by the Boko Haram terrorists in the North East collapse of buildings due to faulty construction specifications which have compounded man's living standard in Nigeria. As a matter of fact, there abound in our society today constant fuel scarcity, rising inflation, bank distress or failures, retrenchment of workers, hired assassination bids, suicide attempts and committals, plenty divorce cases, character assassination, ethnicity and aggressive tribalism; instability in the polity, cultism in high schools and universities; vandalisation of petroleum pipe lines in the Niger Delta areas and other places and frequent burning of strategic national monuments and buildings, markets and constant religious, political cum ethnic riots and murders resulting into huge loss of lives and property; countless automobile accidents arising from bad roads and reckless driving; and occupational hazards or accidents, to mention a few. There are so much insecurity and risks or risk prone areas in our economic and political environment today that hearts of many Nigerians have been failing them and we really need solutions to these traumatic situations. Hence the undisputable need for the knowledge of risk management techniques and application in these times. Next section will state the objectives of the paper. This is followed by discussion on the risk management concept, methodology adopted for the paper; then definitions of risk and uncertainty and reaction to risk. Classification of economic risks and uncertainties were carried out in the next section and then discussed what risk management is and ended up with recommendations and conclusion in the final section. Aims and objectives of the paper The aim of this paper therefore is to specifically address the following objectives: Abstract No doubt many Nigerian people do face a lot of risks and uncertainties today. These manifest in form of economic depression HIV/AIDS infections/epidemic and its resultant impact on per capita income, high unemployment rate and insecurity of jobs, lives and property, high inflation rate, corruption and lack of trust in economic and business relationships among Nigerians, just to mention a few. Upon all these, people are daily seeking for a way out of all these or at least to minimize their scourge by way of employing suggested risk management approaches to tackle these problems. This paper therefore employs descriptive statistic and fundamental analysis approach to define and classify risks into five categories; and then discusses four steps to follow in risk management decision making such as: risk discovery and identification, measurement, decision on the alternative methods to adopt to achieve specific objectives and establish means for effective implementation of the decision made. These are suggestions to be applied by the Nigerian manager at all levels of management and their economic implications for the economy generally. Based on the results from the discussions of the paper, it is recommended for policy adoption and implementation, the followings: (i) proper knowledge of each risk type before its right application, (ii) appropriate application of steps to be followed in risk management to get desired results and (iii) finally, there is urgent need to cultivate awareness of risk management, avoidance and insurance in the mind of Nigerians at all levels of our national life in the 21st century. a. Explain what risk management is all about and classify risks and uncertainties. b. Show how risks could be eliminated or at least minimized in our political, social and economic environments through committed application of basic steps in risk management decision-making and effective use of tools of risk management. c. Make recommendations on the need for imbibing risk management culture and or techniques that will positively influence the Nigerian policy makers' decisions. The risk management concept The quest for security is the eternal concern of man. The history of civilizations reveals how individuals, groups, and even nations have expended their resources and energies toward the satisfaction of an insatiable appetite for security. Besides history discloses how time after time such plans temporarily succeeded but eventually went awry. The pursuit of security is an important part of the total pursuit of knowledge and truth, the main preoccupation of all man's intellectual efforts. In western civilizations the philosophy of government has been to create a society in which the individual has comparative freedom and at the same time greater responsibility in making economic decisions and finding his own economic security. Within this system profits and losses become the acid test of an individual's or a firm's performance. In the Nigerian society, the expectations should not be different. Yet developing economies are still struggling to achieve this economic security. Therefore the business manager must constantly search for ways to improve economic decisions. All available resources, both physical and human must be directed toward isolating the key variables that affect the attainment of the present and future goals of the firm. Cause-and-effect relationships must be ferreted out of the maze of interactivity confronting management, both within and outside the firm. The problem is not new; it is merely a part of a much older and larger problem. Throughout history man has devoted his energies and resources to improving his decisions or predictions of the future. The prophet, philosopher, and "scientists" of early civilization used rather crude research tools and frequently relied on limited observations for establishing rules or principles to guide future acts. These are, of course, risk management techniques on their own merits. However, modern scientists and business persons have deduced from general principles what would happen in a particular case; they have also used induction to generalize on the basis of experimental evidence. Those who make decisions in the field of business management have not been limited to anyone of those methods of problem solving, over the years they have employed, to some degree, all these methods as well as others: intuitive deductive activist (for example, planning) and imaginary. Experts still disagree about the best method to employ in a given set of circumstance. A family, like a business must plan for its future and manage its affairs in such a way as to avoid severe economic losses. The breadwinner, the homemaker, and the children in, say, American economic system, are entitled to spend and invest their resources with considerable freedom as compared with the limitation of individuals in other societies (Nigeria inclusive), but at the same time they are perhaps subject to greater responsibility and to many important economic risks incurred on their changing environment. What America is to the whole world today is presumably what Nigeria is to other African nations, by way of comparison. Methodology We used descriptive statistic and fundamental analysis approach to define and classify economic risks and uncertainties into five broad categories of poverty, liability or personnel risks; physical, social, or market risks; pure and speculative risks; static or dynamic risks and fundamental or particular risks. Data used in the paper were sourced mainly from secondary sources such as textbooks and journal articles. Paper equally made use of University of Abuja Library and other library sources like the Central Bank of Nigeria (CBN), Nigerian Deposit Insurance (NDIC), and NICON Insurance, all in Abuja, to gather data for its analysis. Definitions of risk and uncertainty and reaction to risk Defining risk is not an easy task, as it is evidenced by the numerous definitions, which have been offered by many scholars over the years. Risk is defined as the subject of insurance or the chance of loss (The Dictionary of Insurance Terms, 1994). According to most insurance textbooks, risk is uncertainty with respect to financial loss. Consequently, any definition in this paper is bound to be somewhat arbitrary. Risk and uncertainty Risk will be defined in this paper as objective doubt concerning the outcome in a given situation. It is the doubt a person would have concerning the future outcome even if he knew all the possible outcomes and their probability or chance of occurrence. Specifically, the doubt concerning the outcome which remains in spite of a person's knowledge of the possible losses and their probabilities is his risk. Uncertainty, on the other hand, is subjective doubt concerning the outcomes during a given period. In other words, uncertainty is the doubt, which exists whether or not one knows all the possible outcomes and the probability of their occurrence. Risk is the same for all persons under a given set of circumstances; uncertainty varies among persons and depends on the information at their disposal and their ability to use this information to estimate the risk. If the estimate is perfect, uncertainty equals risk. Reaction to risk The reaction of a firm, family, nation or other individuals or group to risk depends on uncertainty, not risk; it also depends on other factors such as the ability of the persons facing the potential loss to bear possible reverses. Property liability and personnel risks The first method classified risks according to the type of potential losses. a. Property risk exist when property in which the firm or family has a financial interest, other than a liability interest, may be damaged, destroyed, reduced in value, or lost. For example, property risk exists when a building may be destroyed by fire or when value of a business may be reduced by a change in government purchase of its product. b. Liability risk exists when the firm or family will be held legally responsible for property or personnel losses suffered by others. For example, the owner of an automobile for injuries suffered by a pedestrian or a business may have to pay the medical expenses of an injured workman. c. Personnel risk exists when the firm or family may suffer a loss to their persons. For example, the family may fear the possible unemployment of the breadwinner, or a firm may fear the death of a key engineer or salesman. Physical, social and economic risks The second method classifies risk according to the cause or origin of the loss. In his classic book on all forms of risk bearing, Hardy, has described five types of risks classified according to their origin. a. Risk of destruction of property through the physical hazards of nature, such as storms, a flood, or a fire. b. Uncertainties in the.production process; such as variations in the strength of materials or the effectiveness of labour. c. Social risks caused by deviations of individuals conduct from what is expected such as theft, of negligence, and by the impossibility of predicting the behaviour of social groups, such as strikes, riots, wars, and tax reforms. d. Risks caused by the failure or -inability," of individual to use knowledge, which is accessible to them or their competitors, such as failure to use market research information-. e. Market risks, such as price reductions between the dates of purchase and sale of commodities. Mowbray & Blanchard is responsible for the classification of risks as pure or speculative. A pure risk exists when there is a chance of loss but no chance of gain. For example, the owner of an automobile faces the risk of collusion the owner naturally does not gain. On the other hand, a speculative risk exists when there is a chance of gain as well as a chance of loss. For instance, expansion of an existing plant involves a chance of loss and a chance of gain. Pure risks are always distasteful, but speculative risks possess some attractive features. Willet divided risks into static risks and dynamic risks. Static risks are "connected with losses caused by the irregular action of the forces of nature or the mistakes and misdeeds of human beings". They would be present in an unchanging economy. Contrariwise, dynamic risks are associated with changes, especially changes in human wants and improvements in machinery and organization. Static losses usually result in a loss to society, dynamic losses generally do not. A static loss usually affects directly a few individuals at most, while dynamic losses have more widespread effects. Kulp has distinguished between fundamental risks and particular risks. According to him, fundamental risks are group risks, impersonal in origin and effect, and at least for the individual, unpreventable; whereas particular risks are personal in origin and are readily controlled. Examples of fundamental risks are those associated with uncertainties, inaccuracies, and disharmonies in the economic system; risks associated with major social and political changes, and risks associated with extraordinary natural disturbances such as droughts and tornadoes. Examples of particular risks are the risks of death or disability from non-occupational causes, the risk of property losses by such perils as fire, explosion, theft, and vandalism, and the risk of legal liability for personal injury or property damage to others. Particular risks are always pure risks, whereas fundamental risks induce pure and speculative risks. What is Risk Management? At this juncture, we shall consider what risk management entails in a modern economic system. The element of risk pervades all levels of management decisions in a firm or a family. In a firm, therefore, the function of risk management describes properly the functions of all business managers. In this broad perspective, risk management may be defined as the minimization of the adverse effects of risk at minimum cost through its identification, measurement and control. Risks handled by risk mangers Defining the types of risks handled by risk managers is not easy task. The risk manger is responsible for most but not all static risks. The prospect of certain losses to the firm's own product as a result of faculty processing by employees is an example of a static risk for which the firm looks to other departments for correction. The risk manger may, on the other hand, be concerned with a few dynamic risks, such as the inability to collect accounts receivable because of a business decline. Besides the risk manager is concerned with most pure risks. He is not concerned with speculative risks except to the extent that the creation of speculative risks forces him to face certain pure risks; for example, the acquisition of a new plant creates a potential fire loss. On the other hand, some pure risks are ordinarily handled by risk managers, such as the probability that a strike will curtail business operations (though this example is not as apt as it used to be). Strike insurance is now available on a limited scale, and the risk manager may be asked to arrange for this coverage or the possibility that some technological change will put the firm out of business. The four steps in risk management decision making Most authorities in this field of study agree that decisions in the risk management area should ordinarily be made by following four specific steps because of their tested effectiveness. They include: A. Procedures and communications should be established throughout the organization to allow for a complete inventory and discovery of the potential (pure) risks that may arise in the activities of the business firm or family. Risk discovery is the first and perhaps the most difficult function that the risk manager or administrator must perform. B. After identification of risk-s, the next important step is the proper measurement of the losses associated with these risks. This measurement includes a determination of: i. The probability or chance that the losses will occur ii. The impact the losses would have on the financial affairs of the firm, should they occur and iii. The ability to predict the proportion of losses that will actually occur during the budget period. 1. Once the risk is identified and measured, the various alternative solutions or tools of risk combination of tools to be used in attacking the problem. In this respect, the risk administrator must establish the costs of handling his potential losses through alternative methods, including the use of insurance. In selecting the proper tool, the risk manager should consider the present financial position of his firm or family, its overall policy with reference to risk management, and its specific objectives. 2. After deciding among the alternative methods of risk treatment, the risk administrator and perhaps the appropriate management group must establish means for effective implementation of the decisions made. If insurance is to be purchased, shopping the market for adequate and reasonable rate, and the selection of the insurer are part of the implementation process. Recommendations and Conclusion In the paper, we have tried to establish the need for risk management in both political and business environment of Nigeria. We equally defined what risk management entails and afterward-classified economic risks and uncertainties. Four steps to be applied in risk management decision taking were also discussed here for your appropriation. Common opinion in Nigeria today revealed that most people do not take serious the issues of risk management despite existence of deluge of risks in our economic system. Being so, we recommend that henceforth, Nigerian business managers and administrators alike imbibe the principles and tools of risk management. By so doing we would have minimized most of the risks and uncertainties (costs) that do impact negatively on our return on investment (ROI) be it at private sector or public sector levels. We therefore conclude this paper by saying that there is an urgent need for the culture of risk awareness and elimination in our society. When this is fully entrenched in the minds of the Nigerian people, it will surely have positive implications for our ailing economy in the 2000.