Datasets:

iNeil77

/

pseudo-mini-pile

like 4

(1) Field of the Invention The invention relates to power conservation. More specifically, the invention relates to reduction of power consumption in a low power mode. (2) Background Reduced power consumption is generally regarded as a desirable characteristic, but in some power critical applications, reducing power consumption may become essential. Certainly, in mobile devices reduced power consumption will increase battery life and permit longer operation in the mobile environment. Additionally, certain industry specifications require power consumption at a very low level. For example, the Universal Serial Bus (USB) Specification Version 1.0, published Jan. 19, 1996, mandates that bus powered devices draw no more than 100 milliamps after power on, 500 milliamps after enumeration, and no more than 500 microamps when the device is in sleep mode. As USB devices have become increasingly complex, the entire sleep mode power budget has been consumed by the central processing unit (CPU) in the USB device. This leaves insufficient power budget to supply other auxiliary circuitry, including, but not limited to, powering the memory for the processing unit, and any other functionalities. The power problem is exacerbated by the fact that the CPU in sleep mode must be able to awaken rapidly in response to a wake-up signal. Thus, the CPU cannot be wholly disconnected from the power supply during sleep mode. Prior approaches attempting to permit compliance of USB devices with the sleep mode power budget have generally failed. One attempted solution is to have the CPU turn off the auxiliary circuit responsive to receiving the go-to-sleep signal from the bus. Unfortunately, this creates problems in fetching the sleep instructions, as the memory for the CPU is often included in the auxiliary circuitry. Moreover, a greater problem arises when the wake-up signal arrives and the auxiliary circuitry is not yet powered. As a result, the CPU is unable to fetch the instructions needed to appropriately respond as required. Other attempted solutions have generally failed to permit the devices to meet the 500 microamp sleep budget. This problem has been sufficiently vexing that many manufacturers have been lobbying the USB Standards Committee to increase the power budget for sleep mode of USB compliant devices. In view of the foregoing, it would be desirable to provide a way to satisfy power budgets in extremely power-constrained systems, while still permitting acceptable response time and transitioning in and out of a low power mode.

Formation Mechanism of Polymeric Fullerenes Abstract The energetics and reaction paths of polymeric forms of fullerenes have been investigated by means of calorimetry experiment and semiempirical molecular orbital calculations. Energy schemes of three types of single phase C60 polymers i.e., dimer, one-and two-dimensional polymers have been experimentally established. This energy scheme, as well as the dramatic difference in experimental conditions for the polymerization and the depolymerization, is reasonably explained by a calculated energy contour map taking the distortion of fullerene molecules into account.

New reports show U.S. companies are cutting jobs as employers try to cope with the slowing U.S. economy. A private employment firm, ADP Employment Services, says U.S. companies cut 25,000 jobs in November - the biggest reduction in payrolls since 2001. A separate research firm, Challenger, Gray and Christmas, says the financial sector and the automotive industry planned the most job cuts in November as worldwide consumer demand weakened. U.S. bank Citigroup announced the most cuts, saying it plans to reduce its payroll by 52,000 employees. The labor cuts correspond with a contraction in the U.S. service industries, which include banks, hotels, airlines and restaurants. A trade group, The Institute for Supply Management, says its index of business activity for the U.S. services sector fell to its lowest point since records began in 1997. Meanwhile, a report from the U.S. Labor Department shows worker productivity declined at a slower pace than expected, indicating U.S. companies may be able to stay profitable with fewer employees.

Sero Surveillance Study to Monitor the Trends of SARS COV2 Infection in the Pediatric Population of the Goa State 1,2 Department of Pathology, North District Hospital, Mapusa Goa 403507 3,4 IDSP, DHS Goa *Corresponding Author Dr Varsha Munj Senior Pathologist, Department of Pathology, North District Hospital, Mapusa Goa 403507 Abstract Background: As there are concerns raised of anticipated 3 rd wave of Covid19 to hit the state of Goa around August-October 2021 wherein pediatric population is likely to be affected, conducting pediatric population based sero-surveillance will help to estimate and monitor the trends of infection that will guide the strategy for making further decisions. Materials and Methods: The sero -epidemiological investigation for COVID 19 infection is a pediatric population based cross-sectional study (5-11years and 12-17 years). The sample size was calculated for each of the age group and strata (District).480 samples were analysed by serological test to determine SARS COV2 antibodies (quantitative spike proteins). Results: 480 children were considered for the study as per the survey design. 319 were tested positive for SARS COV2 antibody.The overall seroprevalence of SARSCOV2 antibody in children (5-17 years) of Goa state is 66.40% with North district (68.33%) and South district (64.58%). 54% were females and 46% were males. 57% of children were in the age group of 5-11 years whereas 43% were in the age group of 12-17 years. 13% of children presented with mild to moderate symptoms while 87% of children were asymptomatic. The seroprevalence was similar in rural and urban areas of Goa. Betki(North District) had positivity of only 29.17% and Canacona and Ponda (South District) had positivity of 37.5%. Conclusion: SARS COV2 seropositivity in children 5-17 years is higher in Goa state. Vaccine priority can be given age group of 11-17 years. There is heterogenecity in distribution of cases indicating possibility of further waves. Introduction Severe acute respiratory syndrome Corona virus 2 (SARS -COV2) was first reported in China in 2019 and has emerged as a pandemic which has spread to more than 200 countries 1. SARS COV 2 has been reported in all age groups 2. However, http://jmscr.igmpublication.org/home/ ISSN (e)-2347-176x ISSN (p) 2455-0450 DOI: https://dx.doi.org/10.18535/jmscr/v9i9.25 there have been far fewer confirmed reported cases of Covid 19 in pediatric population 3. The infection of SARS COV2 appears to have milder course in children than adults. Most infected children presented with mild symptoms or were asymptomatic. However, MIS-C (Multi -system Inflammatory Syndrome in Children) may present weeks after a child is infected with SARS -COV2 where the child may be asymptomatic or may have been infected from an asymptomatic contact or care giver and is unaware about the infection status 4. As there are concerns raised of anticipated 3 rd wave of Covid-19 to hit the state of Goa around August-October 2021 wherein pediatric population is likely to be affected, conducting pediatric population based sero-surveillance will help to estimate and monitor the trends of infection that will guide the strategy for making further decisions. Many of the Covid -19 pandemic affected countries have initiated community based serosurveillance screening strategies done on large scale to quantify the burden of Covid -19 by using serological tests to detect Covid-19 antibodies by ELISA and CLIA methodology and rapid tests (Lateral Flow Immunoassays). Most of the studies are targeting adult population 5,6. In a study conducted in Delhi, the sero-survey of SARS-COV-2 infection among the pediatric population was 55.7% 7. A separate study conducted in pediatric population of Mumbai seroprevalence was 51.18% 8.Very less data is available with regard to sero-prevalence in pediatric population of Goa. The surveillance program will give a baseline prevalence of seropositivity in paediatric population in Goa. It will help to take decisions with regards to 1) vaccination of this group.2) opening up of schools 3)guide the state to build up its capacity 4) customize to design and implement appropriate measures. Study Population (Methodology) The proposed study is a pediatric population based cross sectional study. Serological investigation to determine SARS COV2 antibodies was carried out in age group of 5-11years and 12-17 years in Goa state. The sample size was calculated for each of the age group and strata (District) separately (table 01) The required sample size for each stratum (District) was 460, based on the seroprevalence of 50%, with a desired precision of 6.5%, with 90% confidence. Design effect of 2 and 10% non response rate was calculated. 2024 households visited in each district. Goa has two districts, with 25 PHC, 4 UHC, and 6 CHCs. The PHC, CHC, and UHC are considered as clusters (primary sampling units). From each district, 7 PHCs, 2 CHCs and 1 UHC was selected according to systematic probability proportional to population size. From the selected each of the clusters 202 households will be visited. The household list of each PHC, CHC, and UHC are available. Each of the age group has the same sample size. (Table 02) All personal data is kept confidential and was processed anonymously. Laboratory Evaluation: Laboratory procedures involving sample manipulation was carried out in the Bio-safety cabinet. Validation and Quality Control: SOPs were followed of the Laboratory Serological Testing: Enzyme -linked Immunofluorosence Assay technology for the quantitative determination of Spike protein (anti-S) specific IgG antibodies to SARS-COV-2 in human serum samples by using Fully Automated Vidas R of Biomerieux Clinical Diagnostics. The assay is intended as an aid in the diagnosis of CoVID-19 and to support the study of the immune status of infected patient by providing an indication of the presence of neutralizing IgG antibodies against SARS-COV-2.Simultaneous comparison will be done by using lateral flow assays. Results A prospective study was carried out to determine the seroprevalence of SARS COV2 antibody in children (5-11 years and 12-17 years) in both the districts of Goa at Pathology Department, North Goa District Hospital, Mapusa. A total of 520 children from age group of years participated of which 480 were considered for the study as per the survey design. Refer Table 3,4 and 5 for results. Of the total 480 children 319 were tested positive for SARS COV2 antibody.The overall seroprevalence of SARS-COV2 antibody in children (5-17 years) of Goa state is 66.40%. The North district of Goa has a higher seroprevalence of 68.33% as compared to South district of Goa which has a seroprevalence of 64.58%. 54% were females and 46% were males. Out of the total positives, 57% of children were in the age group of 5-11 years whereas 43% were in the age group of 12-17 years. Out of the total positives, only 13% of children presented with mild to moderate symptoms of flu, nausea, vague abdominal pain, throat ache and headache. 87% of children who tested positive for antibodies were asymptomatic.21% of children enrolled for the study gave history of exposure with positive person(parents and contacts). 7% of children enrolled for the study gave history of COVID positivity and had no history of any hospital stay. In North district of Goa the samples representing from Betki had positivity of only 29.17% and in South Goa district Canacona and Ponda had positivity of 37.5%. The seroprevalence was similar in rural and urban areas of Goa. Discussion The overall seroprevalence of SARS-COV2 antibody in children (5-17 years) of Goa state is 66.40% which was higher as compared to the study done in Delhi showing a positivity of 55.7% 7 and study done in Mumbai children who had a positivity of 51.18%. 8 The North district of Goa has a higher seroprevalence of 68.33% as compared to South district of Goa which has a seroprevalence of 64.58%. This shows that a higher percentage of pediatric population is exposed to the virus in both the districts of Goa with a preponderance in North District. There is a slightly higher SARS COV2 antibody positivity rate among female children (54%) as compared to male children (46%). Our observations are similar to findings by Puneet et al 9. However metaanalysis has shown higher prevalence in men. 10 A higher seropositivity was observed in children with age group 5-11years (57%) as compared to Short Comings High seroprevalence which appears soothing may have its own short comings as sero-survey results are calculated projections of small sample to huge population. Hence large population based study is needed at repeated interval and the present study may be used as a baseline for the same. 3) There is a ray of hope with regards to developments of Herd immunity in pediatric population however there is no room for complacency with regards to maintenance of COVID appropriate behavior and stressing on proper mask practices. 4) Social, public, religious and political gatherings to be avoided.

John C. McQueen Early career John C. McQueen was born on July 5, 1899, in Carrollton, Missouri, but grew up in Colorado Springs, Colorado. Following the high school, he attended the United States Naval Academy at Annapolis, Maryland, and graduated with Bachelor of Science degree in Mathematics and Engineering on June 3, 1921. McQueen was also commissioned Second lieutenant in the Marine Corps on the same date and assigned to the Basic School at Quantico Base for further officers education. He was subsequently assigned to the First Marine Brigade and sent to Haiti. McQueen was stationed at Port-au-Prince until 1925, when he was appointed commander of the Marine detachment aboard the cruiser USS Cleveland. He subsequently sailed for Nicaragua, where Marine units were tasked with the suppression of the Sandino rebellion. McQueen was appointed commander of the Constabulary Detachment of Guardia Nacional in the town of Estelí. He was decorated with the Nicaraguan Cross of Valor with Diploma for his service in Nicaragua by the president of Nicaragua, José María Moncada. During his service in Nicaragua, McQueen bought a jaguar kitten in the market and named him Bill. Bill sat on McQueen saddle as he led the troops on horseback through the jungles of Nicaragua, and then ran alongside them as he got bigger. He slept at the foot of McQueen's bed until he met his demise (a local farmer's shotgun) when he was caught hunting the neighbor's chickens at night. McQueen was later appointed commander of the Marine detachment aboard the newly commissioned cruiser USS Quincy in June 1936. Quincy was subsequently ordered to Mediterranean waters to protect American interests in Spain during the height of the Spanish Civil War. McQueen and his marines assisted with the evacuation of the foreign nationals to Marseille and Villefranche, France. He returned to the United States in October 1936 and McQueen was assigned to the Division of Plans and Policies at Headquarters Marine Corps. During the summer of 1940, McQueen received special order from Commandant of the Marine Corps, Major General Thomas Holcomb. Holcomb subsequently ordered: "Select a pilot....get a plane...and find us a training center." Major McQueen chose Captain Verne J. McCaul, Assistant Operations and Air Liaison Officer of the 1st Marine Brigade and spent next six weeks with the flying along the East coast. They surveyed the area from Corpus Christi, Texas, to Norfolk, Virginia, until their attention was caught by the 14 miles area by the New River, North Carolina. They recommended the area to the Commandant Holcomb, who evaluated the area as suitable and ordered the construction of the training camp. The training center which was later built, get the name, Camp Lejeune. McQueen and McCaul later received Letters of Commendation from Holcomb for their efforts. World War II During World War II in Europe, Major McQueen was ordered by Commandant, Major General Thomas Holcomb, to Great Britain, where he was assigned to the United States Embassy in London as a military observer. His main task was to consult with British military officers about amphibious tactics and operations and observed training of the British Commandos. McQueen also witnessed the German bombardment of cities and industrial centers and was concerned with the lack of security at U.S. embassy in London. The United States Ambassador to the United Kingdom, John Winant, was impressed by the McQueen's comments and appointed him embassy security officer. His tour of duty in Europe ended in July 1941 and he subsequently returned to the United States. McQueen was subsequently promoted to the rank of lieutenant colonel and assigned to Headquarters Marine Corps in Washington, D.C., where he served with the Division of Plans and Policies under Brigadier General Charles D. Barrett. While served in this capacity, he was involved in the selection for the new marine training ground on West Coast. He and his team subsequently recommended the area of Rancho Santa Margarita, California. Later in summer 1942, Camp Pendleton was built on the place which McQueen designated. McQueen was subsequently appointed director of Intelligence section (M-2) within Division of Plans and Policies in June 1942. He also simultaneously served as an intelligence officer on the staff of the Commander, Amphibious Forces, Pacific Fleet and participated in the planning of the battles of Kiska and Attu and its seizure and occupation. He was subsequently decorated with the Legion of Merit for his service during the campaign. When V Amphibious Corps was activated at the end of August 1943 under the command of Lieutenant General Holland Smith, McQueen was appointed Corps Operations officer. He served in this capacity during the Marianas Islands Campaign and later during the Recapture of Guam in summer 1944 and received his second Legion of Merit, now with Combat "V". When Holland Smith was appointed commanding general of the Fleet Marine Force, Pacific in August 1944, he requested McQueen as his operations officer. McQueen was meanwhile promoted to the rank of colonel and served in this capacity until November 1944. His next assignment was with newly activated 6th Marine Division under Major General Lemuel C. Shepherd, where he relieved Colonel John T. Walker as division chief of staff. The 6th Division later participated in the Battle of Okinawa at the beginning of summer 1945 and Colonel McQueen participated in the battle with distinction. He received his third Legion of Merit for his part in the battle. Postwar service Following the Surrender of Japan in August 1945, 6th Division was sent to Tsingtao, China on 11 October 1945 for the repatriation of Japanese troops. McQueen was decorated with the Bronze Star Medal for his service during the operation. He also received the Order of the Cloud and Banner 5th Class from the Government of China. McQueen was relieved by Colonel Harry E. Dunkelberger on 16 February 1946 and returned to the United States. Upon his return home and two-month leave, McQueen reported as chief of staff at Troop Training Unit, Atlantic at Little Creek, Virginia in April 1946. He remained in this capacity until February 1948, when he has been transferred to San Francisco as commanding officer of the 12th Marine Corps Reserve District. A great highlight of his career came in May 1950, when he was promoted to the rank of brigadier general and subsequently appointed director of Marine Corps public information in September of that year. McQueen was subsequently promoted to the rank of major general in August 1952 and appointed Director of Marine Corps Reserve. He was transferred to California in May 1954, when he relieved Major General John T. Walker as commanding general of the Marine Corps Recruit Depot San Diego. In this capacity, he was responsible for the training of 21,000 new recruits each year. His last assignment came in August 1956, when he was ordered to The Hague as Chief of the Military Assistance Advisory Group to the Netherlands. He succeeded Major General Ray A. Robinson in this capacity. McQueen retired from the military service on 1 July 1958 and was advanced to the rank of lieutenant general on the retired list for having been specially commended in combat. Lieutenant General John C. McQueen died on December 7, 1985, at his home in Menlo Park, California, and his ashes were scattered at sea.

Fundamentals of Presbyopia: visual processing and binocularity in its transformation Background The accommodation has considerable interactions with the pupil response, vergence response and binocularity. The transformation of visual reception processing and the changes of the binocular cooperation during the presbyopia development are still poorly studied. So, the regularities of visual system violation in the presbyopia formation need to be characterized. This study aims to reveal the transformation of visual reception processing and to determine the role of disturbances in binocular interactions in presbyopia formation. Methods This study included 60 people with emmetropic refraction, uncorrected distance visual acuity 1.0 or higher (decimal scale), normal color perception, without concomitant ophthalmopathology. The first group consisted of 30 people (from 18 to 27 years old) without presbyopia, the second cohort comprised 30 patients (from 45 to 55 years old) with presbyopia. The eyeball anatomy and optics were evaluated using ultrasound biomicroscopy, aberrometry, and pupillometry. The functional state of the visual system was investigated under monocular and binocular conditions. The registration of the disparate fusional reflex limits was performed by the original technic using a diploptic device which facilitated investigation of the binocular interaction under natural conditions without the accommodation response, but with the different vergence load. The disparate fusional reflex was analyzed using the proximal and distal fusion borders, and the convergence and divergence fusion borders. The calculation of the area of binocularity field was performed in cm2. Results The presbyopia formation is characterized by a change in an intraocular anatomy, optics, visual processing, and binocularity. The processes of binocular interaction inhibition make a significant contribution to the misalignment of the visual perception. The modification of the proximal, distal and convergence fusion borders was determined. It was revealed that 87% of the presbyopic patients had binocularity shortage, whereas the reduction of binocularity field area in extreme grade was seen in 6% of cases. Conclusions The presbyopia formation is accompanied by a significant reorganization of the visual system activity and by the creation of the new visual processing interactions. These data may be useful in presbyopia surgery. Background According to the International Classification of Diseases (ICD-10, version: 2010) presbyopia belongs to the class of refraction and accommodation disturbance and is slow, age-related and irreversible accommodation decline. Currently, over 1.7 billion people in the world are afflicted with presbyopia. The problem appears in people at the age of 40 -45 years, which is the period of maximal professional and creative activity involving analyses of a significant volume of visual information. The problem of presbyopia correction is an actual task, but in spite of the active introduction of advanced methods, the question about adequate presbyopia correction remains open and unsettled. For the compensation of accommodation deficit, different variants of a multifocal optical system creation have been described [1,. But the creation of a multifocal monocular optical system or anisometropic binocular optical system is closely associated with the adaptation processes, which are very complicated in some patients. The multifocal monocular optical system does not have any physiological analogs. Regardless of the multifocal IOL model, there is always a proportion of patients who complain of the visual dysphotopsia as blurring, misting, "holographic" view, 3D -view. In 5% of cases, this dysphotopsia syndrome becomes rigid and is an indication for IOL explantation. The reasons for the visual dysphotopsia are not clear enough. The explantation of multifocal intraocular lens is debated as an "army method of refractive surgery" and is about 3 to 10% according to the data by different authors. Most of the presbyopia theories consider the intraocular changes purely. The presbyopia is mainly viewed as an accommodation decrease determined by the reduction of lenticular elasticity and changing lens suspension apparatus. At the same time, the accommodative response is a part of the near synkinetic reflex. The accommodation has significant interactions with the pupil response, vergence response, and binocularity. However, based on the theory of functional systems, the loss of useful component in the body's activity is accompanied by the measures to compensate or to adapt for it. Therefore, the reduction of the accommodative response that underlies the presbyopia development must be inevitably accompanied by the imbalance among the components of the near synkinetic reflex. How does the accommodation decrease influence the ocular motor forces which are responsible for the stable constant visual image? This problem has not been solved yet. The changes of the binocular cooperation during the presbyopia development are still poorly studied. According to our previous studies, there is some deficit of binocular cooperation in age-related accommodation loss. Besides, the aging-induced accommodation decrease is strongly linked with other physiological aging processes and the degradational changes of sensory neurons. The normal aging is accompanied by the visual acuity decrease, contrast sensibility decrease, color perception changing, stereoacuity decrease. The age-related reduction of stereovision is marginally correlated with fusional ability decrease. But what remains unclear is the role of binocular disorders among mechanisms underlying presbyopia pathogenesis. Therefore, comprehending the binocular interaction changes in people with presbyopia is a critical issue, and the regularities of visual system violation in the presbyopia formation need to be characterized. The purpose of this study is to examine the transformation of visual reception processing and to determine the role of binocular interactions disturbance in presbyopia formation. Subjects The study adhered to the tenets of Helsinki Declaration and was approved by the Institution Research and Ethics Committee (protocol number 8/13 from 25/11/2013). All patients were adequately informed and signed a consent form. Of the 60 people examined, the first group consisted of 30 people (from 18 to 27 years old) without presbyopia, and the second group comprised 30 patients (from 45 to 55 years old) with presbyopia. The criteria for inclusion in this study comprise the presence of emmetropic refraction (i.e., the spherical equivalent of cycloplegic measurements from +0.25 D to −0.25 D), uncorrected distance visual acuity of each eye 1.0 (decimal scale) or higher, normal color perception, ophthalmopathology absence. All patients of the second group complained of the insufficient near vision, and they made use of the glasses. Considering that binocularity correlates the heterophoria value and interpupillary distance, the exclusion criteria comprised a heterophoria degree greater than five prism diopter, the pupillary distance less than 62 mm and greater than 64 mm. Measurements All patients had a full ophthalmological examination including the evaluation of 80 parameters of eye anatomy, visual processing, and binocularity. Assessment of the eye anatomy and the physiological optics The refractive error was the average spherical equivalent of five cycloplegic measurements taken with an autorefractor/ keratometer (KR8800, Topcon, Japan). All cycloplegic measurements were made 25 min after the administration of 1% tropicamide (2 drops during 5 min interval twice). The other investigations were performed 7-10 days later. The amplitude of accommodation (AA) was measured using a minus lens method. The subjects were asked to fixate N8 target at a distance of 40 cm. Then minus lenses were introduced in 0.25 D steps until the patient reported the first sustained blur that could not be cleared by the further conscious effort. This procedure was done for each eye first monocularly and then binocularly. The total AA was estimated as the endpoint minus lens which was possible to see the target at 40 cm under binocular conditions. The AA measurement in people with presbyopia was done using the near addition lens. The axial length, lens thickness, anterior chamber (AC) measurements were made with the help of an ultrasound biometer (AL-3000, Tomey, Japan). An average of three measurements for each parameter was used. Ultrasound biomicroscopy (UBM) measurements were made using HiScan 2000 (Optikon, Italy), and UBM was done in the supine position as described by C. Pavlin and F. Foster. Images from the iris root to the pars plicata zone were obtained in 12 o'clock direction. The ciliary body thickness in cross section and the length of anterior portion Zinn's ligament were examined. The ocular wavefront aberration across a 3 mm zone in the pupil and the pupil diameter (under photopic and mesopic conditions) were obtained using the principle of automated retinoscopy (OPD-Scan, NIDEK, Japan). The anterior chamber volume, iris and lens configuration, corneal aberrations, lens light transmission, were fixed using Pentacam (Oculus Optikgerte GmbH, Germany). Assessment of visual processing under monocular conditions The distance visual acuity and the near visual acuity were measured with logical geometric scale Bailey-Lovie (logMAR). Other examinations included evaluation of the contrast sensitivity at spatial frequencies 3, 6, 12, 18 cycles per degree (CSV-10000E, VectorVision, USA), the threshold of light sensitivity (EP3000, Tomey, Japan), the flicker fusion threshold, the amplitude and implicit time of maximal electric retinal response, and the amplitude and implicit time of visual evoked potentials (EP1000, Tomey, Japan). Binocularity assessment For this purpose, the data reflecting the different levels of binocular interaction were systematized. These include the near physiological diplopia, the stereovision test Lang I&II, and the binocularity field (spatial limits of disparate fusional reflex). To induce the physiological diplopia, each patient was asked to look into the far distance (5 m). Then a nearlying target (in the distance 10 cm, i.e., not in the horopter curve) was presented. The presence (yes/no) of double virtual objects was fixed. In the case of physiological diplopia, the registration of the disparate fusional reflex limits was performed. For this, a diploptic device (AVIS 01, Krasnogvardeec, Russia) was used, which facilitated investigation of the binocular interaction under natural conditions without the accommodation response, but with the different vergence load. The first point of the measurement is 40 cm. Then the double targets were moved inwards until the virtual stereo image was observed ( Fig. 1). It was a point of binocularity without vergence load. The change in the distance between the centers of the double near lying tests (p) and the distance from the eyes to the targets (n) while maintaining of the virtual binocular image perception makes it possible to define the fusional reflex limits in space. Then the targets were moved increasingly inward and outward to force the vergence response (Fig. 2). There were some points of maximum convergence and divergence. The patient reported all the visual images which were recorded. The disparate fusional reflex was analyzed using the following parameters: 1. The proximal fusion border (PF) and distal fusion border (DF) were determined while the double targets were approaching and moving away (Fig. 2). The difference between these parameters corresponds to the length of the binocularity field. 2. The convergence fusion border (CF) and the divergence fusion border (DivF) were determined with the help of the decrease and the increase of the distance between the double targets (the point of the measurement is 40 cm from the eyes). The Fig. 1 The perception of virtual binocular visual image difference between these parameters corresponds to the width of the binocularity field. 3. Finally, we performed the calculation of the binocularity field area (A) in cm 2 (Fig. 3). Statistical analysis All data were analyzed with a spreadsheet application (Statistica ver. 8.0; StatSoft Inc., USA). The data were represented as the mean value ± standard deviation (Mean ± SD). The Shapiro-Wilk test was used for assessing of normality distribution. The statistical differences in measured values were analyzed using a t-test. The critical level of significance (p) upon the examination of statistical hypotheses was 0.05. The comparison analysis, the Pearson correlation analysis, and the logistic regression models were done. The Pearson correlation coefficient with absolute values equal to or greater than 0.7 with p < 0.001 was accepted as the close relation. The discriminant function analysis (DFA) was used for the selection of the analytes that maximally discriminated the studied groups. The DFA was built in a step-wise manner after direct standardization. The final discriminatory power of each analyte was characterized by a partial Wilk's Lambda coefficient; 1.0 (no discriminatory power) to 0.0 (perfect discriminatory power). The Mahalanobis distance D 2 between centroid values for each group was measured. Results The baseline features of the study population were summarized in Table 1. The mean patient age was 22.3 ± 3.2 years in the first group, and 52.4 ± 2.2 years in the second group. The groups did not differ in gender, spherical refraction equivalent, eye globe axial length, oculomotor status. In the first (control) group, the mean accommodation amplitude was 6.93 ± 1.12 D (minimum Intraocular anatomy and optics The presbyopia formation is characterized by a change in an intraocular anatomy. Significant differences were detected in the anterior-posterior size of the lensfrom 3.73 ± 0.23 to 4.41 ± 0.21 (p = 0.0001). The optical density (light transmission coefficient) of the lens increased in the nuclear area from 15.5 ± 1.2 to 26.6 ± 3.4% (p = 0.0001), and in the cortical layersfrom 9.1 ± 0.9 to 10.8 ± 1.3% (p = 0.03). The change of the intraocular anatomy was expressed as: the decrease of the ciliary body thickness (in the inner top projection) from 0.82 ± 0.10 to 0.63 ± 0.11 mm (p = 0.001), the increase of the distance between the trabecula and the ciliary processes from 0.79 ± 0.10 to 1.02 ± 0.11 mm (p = 0.001), and the shortage of the front portion Zinn ligament length from 1.23 ± 0.31 to 1.04 ± 0.26 mm (p = 0.002). The variation of both static and dynamic components of the optical physiological system was established in the eyes of presbyopic patients. This was evidenced by a significant increase of the total root mean square wavefront errors (from 0.13 ± 0.04 to 0.17 ± 0.05 m in pupil diameter 3 mm, p = 0.0001) and the corneal root mean square wavefront aberrations. In patients with presbyopia, an increase in the Zernike coefficients of the corneal spherical aberration Z 4 0 from 0.17 ± 0.05 to 0.23 ± 0.06 m (p = 0.0001) was observed. Also, a significant decrease of the pupil excursion was found. The photopic pupil diameter decreased from 3.81 ± 0.76 to 3.35 ± 0.78 mm (p = 0.0001) whereas the mesopic pupil diameter decreased from 6.47 ± 0.56 to 5.50 ± 0.94 mm (p = 0.0001). Monocular visual characteristics In the next step, a comprehensive study of the sensory activity of the visual system was carried out. The patients with presbyopia had a decrease not only in accommodation amplitude and uncorrected near visual acuity but also in the most of the visual reception parameters ( Table 2). A significant decrease of the contrast sensitivity in low and high spatial frequency ranges, the average values of the b-wave ERG maximum amplitude, the flicker fusion threshold and an increase in the a-wave and b-wave ERG maximum latency were found in patients with presbyopia. Binocularity The change in the perception Lang stereo tests was not significant. A study of binocular cooperation showed an inhibition of the near physiological diplopia in 20% of the patients with presbyopia. There were multiple changes. These include a significant distortion of the binocular interaction zone with the reduction of total binocularity area, shift in the space towards the near focal point, and the fusion neutralization in the convergence zone. The changes in the proximal, distal and convergence fusion borders were determined ( Table 3). The regressions between disparate fusion reflex limits and accommodation amplitude are shown in Fig. 4. In the first (control) group, the mean area of binocularity field was 365.6 ± 45.1 cm 2 (minimum 280 cm 2, maximum 456 cm 2 ). All patients with presbyopia had a decreased fusion ability with the mean area of binocularity field at 174.4 ± 87.7 cm 2 (minimum 48 cm 2, maximum 385 cm 2 ). The regression between binocularity field area and accommodation amplitude is shown in Fig. 5. It is interesting to note that 77% patients with presbyopia had binocular suppression in some grade, while 6% of patients had extreme decrease in the binocularity field area (Fig. 6). Intrasystem interactions The multivariate kinds of statistical analyses were used for the understanding of the vision reception transformation in presbyopia formation. A Pearson correlation analysis of visual system parameters was made. The comparison of the correlation Pleiades (correlations with P-value equal or less than 0.001) within Control and Presbyopia Groups revealed a reduction in the strength of most relationships. The correlation Pleiades are represented in Fig. 7, where the positive correlations were shown as red arrows and the negative correlations as blue ones. It is evident that young people without presbyopia have a much larger number of interdependencies between structural and functional indicators of the visual system than in patients with presbyopia. In young people, 25 close relationships were established, but in patients with presbyopia, there were only 6 such relationships. Instead of destroyed relationships, there was a new correlation between the width of the binocularity field and the implicit time of visual evoked potentials (r = −0,7; p = 0.001). In the forward stepwise discriminant analysis, eight indices were selected for 100% discrimination of studied groups. The matrix of most informative variables for discrimination is represented in Table 4. The separation between groups is not only due to the accommodation state but also to some other significant The results of the discriminant analysis showed that the fusional ability made the high contribution in the separation of two groups. At the same time, the contribution of other sensory parameters in the division was less expressed. Discussion This research aimed to describe the transformation of the visual system functional organization during the presbyopia formation. The results of the study broaden our understanding of the presbyopia mechanisms. It was found that the structural and functional state of the visual system in middle-aged patients with presbyopia is significantly different from young people. The reduction of the accommodation is significant, but not the only sign of visual transformation in patients with presbyopia. The increase in the number of optical errors despite the pupil tour decrease worsens the conditions for the formation of the retinal image. The formation of presbyopia is accompanied by the misalignment of visual sensory processing with varying degrees of functional defect severity (Fig. 8). The decrease of the contrast sensitivity at frequencies below four cpd reflects the amplification of the visual system internal noise (at the level of receptive fields). Whereas the contrast sensitivity at high spatial frequencies is limited with optical parameters (aberrations, diffraction phenomenon or "sampling noise" of the visual image). The patients with presbyopia had the signs both internal noise and sampling noise (noise of signal processing). The change of temporal parameters of the visual system at middle age patients indicates the initial deficiency of interactions between neurons and the signs of the central nervous system fatigue. These changes can be viewed as signs of aging. The new data about binocular visual system activity was established. The processes of binocular interaction inhibition make a significant contribution to the misalignment of the visual perception. The area of binocularity field, where the disparate fusion is possible, was reduced twice. The variation of proximal fusion limit and a decrease of the binocularity field area are more serious than the variation in other sensor parameters. On the one hand, this phenomenon may be a result of age-related changes in the neurons. On the other hand, the inhibition process can be motivated by the desire to liberate the body from the excessive flow of the visual information under the shortage of accommodation. The process of efferent synthesis is an active selection of information aimed to release biologically significant flows and is formed in such integral efferent excitations that are required by the body in a given situation. Our results correspond to the relevant studies. Granger-Donetti revealed that majority of presbyopes had disorders of binocular cooperation in some degree due to a decrease in the slow convergence. The accommodation amplitude decrease is accompanied by the increase of vergence. 8 The Changes of the visual parameters (M) in patients with presbyopia in comparison with indicators of young people movement latency, the reduction of the vergence fusion and the speed of the fast vergence. In this study, we analyzed data in people with the visual system that mostly meets the ideal. Even in this situation, 77% of presbyopia patients had binocularity shortage. In 6% of cases, there were profound signs of the deep inhibition processes. These data are likely useful in presbyopia surgery. The analysis of the surgery results using monovision or multifocal optical strategies in patients with an extreme deficit of binocularity is the next step of research. Conclusions The presbyopia formation is accompanied by a significant reorganization of the visual system activity and the creation of the new visual processing interactions. It was revealed that 77% of the presbyopia patients had binocularity shortage. The overall reduction of binocularity field area in extreme grade was seen in 6% of cases. These data may have implications for presbyopia surgery.

Purification and Studies of Some Properties of Pig-kidney Biliverdin Reductase. Biliverdin reducatase was purified from pig-kidney to homogeneous form through DEAE 52-cellulose, Sephadex G-100 and Procion red HE 3B-Sepharose 4B column chromatography, The enzyme was estimated to be a monomer with a molecular weight of 66kD by SDS-PAGE. A gel staining in basic-pyridine solution of heme was used to identify purified biliverdin reductase. The enzyme utilized NADPH and NADH as electron donors for the reduction of biliverdin to produce bilirubin. The apparent K(m) values for biliverdin were established to be 1.25 &mgr;M with NADPH and 2.52 &mgr;M with NADH.

Formaldehyde inactivation of measles virus abolishes CD46-dependent presentation of nucleoprotein to murine class I-restricted CTLs but not to class II-restricted helper T cells. To induce an MHC-restricted specific CTL or Th response, an antigen must be delivered into the appropriate cellular compartment. We explored the role of CD46 in the presentation of measles virus (MV) nucleoprotein (NP) to murine NP-specific and MHC Class I-restricted polyclonal CTLs and the effect of inactivating MV by uv or formaldehyde. CD46(-)- and CD46(+)-transfected murine cells were used as target cells. After MV infection, only the targets which expressed CD46 were lysed by NP-specific class I-restricted CTLs. When MV was uv-inactivated, NP presentation by MHC class I molecules was retained but could be blocked by fusion inhibitors which block virus cell entry. When MV was inactivated with formaldehyde, NP was no longer presented by MHC class I molecules, although it was still presented by MHC class II molecules to a NP-specific class II-restricted T cell hybridoma. These data show that MV binding to the CD46 molecule is a prerequisite for virus-to-cell fusion and that cytosolic delivery of NP is necessary for presentation by class I molecules. Moreover, formaldehyde inactivation of virus induces the loss of class I-restricted presentation of NP due to selective abrogation of fusion and cytosolic delivery of NP.

SELF CLEANING OF EARS BY VARIOUS OBJECTS CAUSES MULTIPLE DISEASES OF EXTERNAL EAR Objective: To find out the association of multiple diseases of external ear with the use of various objects and cotton buds. Study Design: Case control study. Place and Duration of Study: Ear Nose Throat Department, Pakistan Naval Ship Shifa Karachi, from Jan to Oct 2020. Methodology: All the patients having multiple diseases of external ear were included as cases. Controls were selected from the healthy volunteers from general populations. Results: Total number of subjects in this study was 140 with the mean age 25.11 ± 11.21 years (2-55 years of age range). Out of them, 82 were male and 58 were female. Odds ratio was 16.07. Conclusion: There is a solid relationship of illnesses of external ear with the utilization of cotton buds. This detrimental practice should be incapacitated by featuring the notice by brands owners of cotton bud and safety instructions at different instructive levels. INTRODUCTION Cleaning is good habit and care of the body is the basic right of a human being. Knowledge regarding the cleaning of ears with various objects like cotton bud, match sticks, fingers and hair pins, should be communicated to masses for quality care of ears. Various objects as mentioned are very injurious to the external ears. These objects include cotton buds which were used for quality care of ears initially but the harmful effects were observed in 1972 in New Zealand. 1 The medical professionals in general and otolaryngologist in particularly are writing articles and advising patients to stop self-cleaning of ears with any object. Cotton buds are utilized by grown-ups as well as are generally applied in pediatric populace either by kids themselves or by guardians. Ear wounds started by cotton buds are regularly seen in ENT outdoor facilities. 2,3 Cotton bud and different articles utilization in ear propensity is having solid relationship with neurodermatitis and contact dermatitis of the outer ear. 4 Cotton bud is additionally connected with intense otitis externa, impaction of wax, burst of tympanic membrane and perichondritis in couple of patients. This study was done to find out the association of multiple diseases of external ear with the use of various objects and cotton buds. METHODOLOGY This case control study was conducted at Ear Nose Throat Department, Pakistan Naval Ship Shifa Karachi, from January to October 2020. Acceptance from Hospital Ethical Committee was acquired on a certificate. Consent from the examination subjects were taken prior to filling the questionnaire for study. Non probability convenience sampling technique was used. Inclusion Criteria: All the patients having multiple diseases of external ear were included as cases after being examined and diagnosed by the otolaryngologist. Controls were chosen from the sound volunteers from all-inclusive communities for same number of patients in span of study. Exclusion Criteria: Patients having other diseases of head & neck region, were debarred from the study. Odds ratio of the risk (using of various objects and cotton buds to clean the ears) was calculated. Occurrences of the variables were determined by utilizing enlightening measurements of SPSS version 20. RESULTS Number of subjects in this research were 140 with the mean age 25.11 ± 11.21 years. Minimum age was 2 years and maximum age was 55 years. Out of them, 82 (59%) were male and 58 (41%) were female. The gender distribution by percentage waspresented in Figure. Seventy patients were cases and 70 were the controls. Among the cases, 55 were cotton bud/ different article consumers and among the controls it was 13. Odd ratio was 16.07. Table-I is showing the computation and chances proportion. Table-II Table-III is showing the questionnaire for exposed population of study (n=68). Table-IV is depicting answers to questionnaire by exposed population (n=68). Answers of exposed population used 100% cotton buds, 76.5% for more than 1 year and 100% without knowing the adverse effects. Various objects like fingers, match sticks & hair pins were also used. Out of 46.5% of adults used to clean their children's ears with cotton buds. DISCUSSION This study showed that a person with diseases of external ear was 16.07 times more likely to have been exposed to the risk factor (using the cotton buds) than a person without ear disease. The case control concentrate on this subject was completed by Ahmed et al in Darfur, Sudan 4 few years back and odd proportion determined was. 12 Our study is not only supporting the previous study and showing more association with exposure of cotton buds with ear diseases. All other studies are descriptive and observational studies showing the importance of risk factor and guiding the masses to stop cleaning ears with any object. Gadanya et al, observed thatthere was a high prevalence of cotton bud use for self-e ar cleaning at Aminu Kano Teaching Hospital. This finding had wider implications as doctors 8 were looked upon by the nonprofessional community and other health workers as role models. Khan et al, observed 79.6% use of cotton buds in undergraduate university students for self-ear cleaning. 9 Alshehri et al, conducted study in medical and non-medical students, 10 and the majority (75%) of the participants practiced self-ear cleaning. These studies are different from our study as we performed on patients and control group. We did not include doctors and medical students in our study. For medical professionals, there was a need to plan and guide out wellbeing instruction and advancement procedures that would spread crucial messages to doctors and other health workers in the local area hospitals, that ear cleaning is vital done via able staffs. Adegbiji et al noticed the commonest object, 11 utilized in ear cleaning was cotton bud in 44.5%. Different articles were digit of hand, plume and key. In our investigation the commonest object was cotton bud, then finger and match stick. Adegbiji et al, additionally led concentrate on children, 12 and noticed the most well-known justification ear cleaning was because of individual cleanliness. Ear cleaning was done in 57.1% of the children by their mothers and cotton bud was the most well-known article. In our investigation, 45.6% of youngsters were cleaned by their guardians by cotton buds. There is an overall conviction that there isn't anything incorrect with utilizing cotton buds, and this disintegrates the propensity for utilizing cotton buds essentially to reduce manifestations, for example, itching, 13 expulsion of filth and maybe even with statements of noticed advantages. Tingling ears were the commonest reason, from neurodermatitis and otitis externa of the ear by different investigations. 4,14 The wax evacuation was the main justification utilizing cotton buds revealed from different studies. 15,16 Our research did not endorse tingling and wax expulsion as the significant reasons of cotton bud use in our results as in previous studies. A survey revealed that majority of cotton bud users was uninformed about its harmful influences on ear. 17 In our investigation, 100% of uncovered populace was uninformed of the harmful impacts of selfcleaning of ears with cotton buds. Consciousness of cotton-bud related entanglements is a significant general medical problem. Safer strategy for aural toileting and suction may likewise require promoting. 18 Public mindfulness by all sort of media must be carried out to save people from preventable ear diseases. A few examinations uncovered that neurodermatitis, contact dermatitis, wax impaction, perichondritis and otitis externa 19 are connected with the propensity for self-cleaning of ears with cotton buds. Our investigation results showed these diseases along with lacerations of external auditory meatus and foreign bodies' external ear. ACKNOWLEDGEMENT We are thankful to our parents, teachers and friends. LIMITATION OF STUDY The study is case control on the topic and second in the world. More case control studies and cohort studies should be done to highlight the bad ear cleaning habit. CONCLUSION There is a solid relationship of illnesses of external ear with the utilization of cotton buds on the basis of odd ratio. This detrimental practice should be incapacitated by featuring the notice by brands' owners of cotton bud, safety instruction at different instructive levels and short commu-nity mindfulness talks of otolaryngologists at electronic media and web-based media. Conflict of Interest: None.

The Sing dollar has gained more than 6 per cent against the greenback year to date, making it one of the top performing Southeast Asian currencies alongside the Thai baht. However, analysts expect it to give up some of its gains soon. SINGAPORE: The Singapore dollar has been one of the strongest Southeast Asian currencies in 2017, defying expectations with a year-to-date gain of 6.1 per cent against the greenback. But most analysts said this uptrend may have run its course. DBS Bank's senior currency strategist Philip Wee, for one, described the local dollar as being "too strong" as an "oversold" US dollar struggles to find upward momentum despite a favourable backdrop of interest rate hikes. Last month in particular, USD/SGD came under "heavy selling pressure" amid heightened speculation that other major central banks could be joining the US Federal Reserve in exiting their ultra-loose monetary policies, Mr Wee wrote in a report published on Tuesday (Aug 1). As a result, the Singapore dollar nominal effective exchange rate (S$NEER) was lifted into the "strongest quartile of its zero appreciation policy band", which was "inconsistent with Singapore’s lacklustre economic and inflation data". Mr Wee added that the Sing dollar should start to stabilise between 1.3370 and 1.3820 in the second half of 2017. The local currency was last seen trading at 1.3562 per US dollar on Friday (Aug 4), not too far away from Monday's 10-month high of 1.3553. Mr Khoon Goh, head of Asia research at ANZ Banking Group, is also of the view that the Sing dollar's strength has been "stronger than what is warranted by domestic fundamentals". He cited the "subdued" advance growth estimates for the second quarter and a persistent weakness in the local labour market. "We think the S$NEER should be trading closer to the mid-point, which is around 0.8 to 1 per cent weaker than its current levels," Mr Goh added, noting that the Sing dollar will likely weaken slightly in the fourth quarter to hover around 1.38 against the US dollar. For Mr Julian Wee, senior markets strategist for Asia at the National Australia Bank (NAB), the pace of increase in the S$NEER has been "too fast". Apart from weakness in the US dollar, an outperformance of the Sing dollar against its less-liquid regional peers, such as the Malaysian ringgit and the Chinese yuan, also played a part. "I don't think the MAS (Monetary Authority of Singapore) will be concerned about the S$NEER being 1 per cent above the midpoint of its policy band, but they won't be so accepting of another half a per cent increase in the S$NEER this month," he explained. "The pace of increase in July has been quite sharp and a little too fast." Nevertheless, he is holding on to his year-end target of 1.405 for the Sing dollar as he expects the greenback to recover with the Fed poised to increase rates once more and embark on the process of unwinding the US$4.5 trillion of bonds it holds on its balance sheet before the end of 2017. "These are conditions that favour a strong rebound in the dollar, perhaps in the fourth quarter," Mr Wee told Channel NewsAsia. However, Mr Nizam Idris, the Singapore-based head of foreign-exchange and fixed-income strategy at Macquarie Bank, has doubts about a rejuvenated US dollar. Even with the prospects of balance sheet reduction, he noted that the Fed is likely to go slow in that aspect thereby minimising its impact on markets. "I am assuming the Fed will start with a slow pace of around US$10 billion reduction per month before incrementally accelerating the process to US$20 billion in the first quarter of next year." He added: "At US$10 billion per month, I don't think it's going to have any significant impact on the yield curve. So it's fairly limited room for the dollar to rebound unless the balance sheet reduction picks up a gear or two, and that could be a 2018 story." Mr Nizam also does not think that recent gains in the Sing dollar warrant much alarm given that the S$NEER remains within range despite rising as much as 1.2 per cent last month. "Over the last couple of weeks, there were other issues such as a rise in uncertainty over global growth and monetary policies, as well as the political situation in the US. In an environment of high level uncertainty, the Sing tends to benefit somewhat but it's not very far away from the policy mid-point. "As long as the currency remains within the band of +/- 2 per cent, I think it should be free to move within a reasonable range without triggering any undue concern from the central bank," he said. There was, however, one thing that analysts could find consensus on. Despite the Sing dollar's strength, the MAS is unlikely to make changes to its monetary policy approach in October, they said. Mr Nizam pointed out the "significant level of uncertainty" in both the global environment and Singapore's growth prospects. As such, "it's best for MAS to just sit back and let others do the tightening for them", he told Channel NewsAsia. "With Singapore's very open capital account and a managed monetary policy, higher global interest rates will eventually filter through into higher interest rates domestically and that should buy the MAS time to wait out on its (forex) policy," Mr Nizam added. Mr Wee from NAB also thinks that the MAS will maintain its neutral policy stance as it needs to observe the actions of the Fed, particularly the cutting back of the Fed's balance sheet. "If that happens in September as many expect then that makes things easier for the MAS. If it doesn't, then they will have to reckon with the possibility. Either way, they will have keep one eye on that and in that light, they will not want to move prematurely."

The invention lies in the semiconductor technology field. More specifically, the invention relates to a semiconductor chip, a memory module and a method for testing the semiconductor chip. In order to ensure the correct functioning of semiconductor chips, semiconductor chips are usually tested prior to delivery. In principle, there are two different procedures for this. By way of example, it is possible to connect an external test device to the semiconductor chips and to check the semiconductor chips with the aid of this test device. A disadvantage here is that the semiconductor chips each have to be individually contact-connected, which is complicated given the large number of semiconductor chips on a wafer. Furthermore, it is possible to integrate the test circuit into the semiconductor chip and to drive it externally via an interface. This method is known under the designation BIST (xe2x80x9cBuild In Self Testxe2x80x9d) in the semiconductor industry. In some cases, the test circuit also provides for a self-repair of the semiconductor chips. This method is also known as BISR (xe2x80x9cBuild In Self Repairxe2x80x9d). One disadvantage of the integrated test circuits is that they take up a considerable proportion of the area of the semiconductor chip. This leads to an unacceptable increase in the costs for fabricating the semiconductor chips. It is accordingly an object of the invention to provide a semiconductor chip, a memory module and a method for testing the semiconductor chip, which overcomes the above-mentioned disadvantages of the heretofore-known devices and methods of this general type and which provides for a semiconductor chip and, in particular, a memory module which can be tested in a simple manner. It is a further object to provide for a simple test method. With the foregoing and other objects in view there is provided, in accordance with the invention, a semiconductor chip, comprising: a chip body having a first side and an opposite second side; a multiplicity of external contacts including at least one contact on the first side of the chip body, at least one contact on the second side of the chip body, and a connection connecting the contacts and extending through the chip body. By virtue of the looping-through of contacts of the semiconductor chip, these can be stacked on a control chip and be tested jointly with the aid of the control chip. This method is particularly advantageous if a large number of identical semiconductor chips are to be tested. This is the case with memory chips, in particular. It is also conceivable, in principle, to stack a large number of memory chips on top of one another in order thereby to increase the storage volume per memory module. In the semiconductor chips, the data and address lines are looped through from one side of the semiconductor chip to the other side of the semiconductor chip. The looping-through of the data and address lines affords the advantage that the semiconductor chips can be individually addressed via the data and address lines as in the case of a conventional memory module. In accordance with an added feature of the invention, the semiconductor chip is a memory chip and the external contacts are contacts for data lines and address lines looped through from one side of the memory chip to an opposite side of the memory chip. In accordance with an additional feature of the invention, the contact is a looped-through contact for serial data communication. With the above and other objects in view there is also provided, in accordance with the invention, a memory module, comprising a control chip and a multiplicity of memory chips forming memory units stacked on the control chip, and at least one contact looped through from one side of a respective the memory chip to an opposite side of the memory chip. In accordance with another feature of the invention, the memory chips have a multiplicity of looped-through contacts connected to the internal data lines, address lines, or command lines of the memory chip. In accordance with a further feature of the invention, the looped-through contact is a line for serial data communication. In accordance with again an added feature of the invention, the control chip is formed with a given minimum feature size that is larger than a minimum feature size of the memory chip stacked on the control chip. In accordance with again an additional feature of the invention, the memory chips are contained in a wafer composite stacked onto a wafer with the control chips. In accordance with again another feature of the invention, the control chip is configured to control a performance of a functional test of the memory chip. With the above and other objects in view there is also provided, in accordance with the invention, a method of testing semiconductor chips, which comprises stacking the semiconductor chips on a control chip configured for testing a function of the semiconductor chips and forming contact between the control chip and the semiconductor chips with a contact looped through from one side of the semiconductor chips to another side of the semiconductor chips; and testing the function of the semiconductor chips. In accordance with again another feature of the invention, a wafer composite of a multiplicity of semiconductor chips arranged next to one another is stacked on top of a wafer composite of control chips arranged next to one another. In accordance with again a further feature of the invention, test signals supplied by the semiconductor chips are transmitted to the control chip via looped-through data, address or command lines. Also, the test signals may be transmitted via a serial line. In accordance with a concomitant feature of the invention, control signals for initiating and performing a functional test of the semiconductor chip are generated with the control chip and the control signals are transferred to one of the semiconductor chips via the looped-through contact. Other features which are considered as characteristic for the invention are set forth in the appended claims. Although the invention is illustrated and described herein as embodied in a semiconductor chip, memory module and method for testing the semiconductor chip, it is nevertheless not intended to be limited to the details shown, since various modifications and structural changes may be made therein without departing from the spirit of the invention and within the scope and range of equivalents of the claims. The construction and method of operation of the invention, however, together with additional objects and advantages thereof will be best understood from the following description of specific embodiments when read in connection with the accompanying drawing.

IT’S the summer of Jon Stewart. And nothing better symbolizes the ascendancy of Stewart and his “Daily Show” in this political campaign season than the scheduled appearance this Tuesday night of Sen. John Kerry. It will mark the first time that a presidential nominee has made a campaign stop on the Comedy Central late-night show, and the first such late-night appearance for Kerry since he became the Democratic Party’s nominee. His handlers have no doubt gotten wind of recent research that shows an increasing number of young viewers are not only tuning in to Stewart’s nightly (Monday through Thursday) newscast spoof for the comedy, but also for the news. They’re also getting their “news” from the late-night monologues on “The Tonight Show with Jay Leno” on NBC and “Late Show with David Letterman” on CBS, according to two recent studies – one from the Washington-based Pew Research Center for the People and the Press last January and another from the University of Michigan last week. Stewart, of course, makes no claim that his show is a news program, but that hasn’t stopped some members of his predominantly young audience from identifying “The Daily Show” as their primary source of political news. This same group, according to the research, is less likely to read newspapers or watch the so-called “serious” news programs on the broadcast networks or all-news cable channels. Moreover, many of them say they’re so apathetic about the political process that they don’t even plan on voting. “Sen. Kerry is going on the show to appeal to a younger segment of voters who increasingly use shows [such as “The Daily Show”] as a source of political news,” said Nojin Kwak, assistant professor of communications at the University of Michigan who studied the relationship between the late-night comedy shows and the political engagement of younger viewers. Kerry’s appearance comes six days before the start of the Republican Convention from Madison Square Garden – an event Stewart and his “Daily Show” plan on covering nightly as part of Comedy Central’s “Indecision” election coverage. In July, “The Daily Show” scored big with its coverage of the Democratic National Convention from Boston. The show averaged 1.3 million viewers nightly over the four days of the convention. And even more important to this week’s high-profile political guest, “The Daily Show” was an overwhelming favorite among viewers 18-34, averaging about 500,000 viewers in that age group from 11 p.m. to 11:30 p.m. – far more than tuned in to the cable news channels during the same half-hour. For his part, Stewart tends to downplay the role his clearly satirical show is now playing as a source of news. And more recently on Ted Koppel’s “Nightline” July 27 on ABC, Stewart was even more insistent that viewers regard his show as a comedy show, and not a news broadcast.

A Model of the Perceptual and Conceptual Processes in Graph Comprehension The article proposes that graph comprehension emerges from an integrated sequence of several types of processes: pattern-recognition processes that encode graphic patterns, interpretive processes that operate on those patterns to retrieve or construct qualitative and quantitative meanings, and integrative processes that relate these meanings to the referents inferred from labels and titles. The model is supported by 2 studies that examine the pattern and durations of eye fixations as a person interprets line graphs or answers questions about line graphs that vary in type and complexity. One implication is that graph comprehension might be more accurate and more complete if the graph's format were changed or the audience were educated to lessen the burden of the inferential, interpretive, and integrative processes. In this article, we propose a model of the comprehension of line graphs that emphasizes the close interaction between conceptual processes, such as interpreting labels and scales, and perceptual processes, such as encoding and interpreting the slopes and patterns of the lines themselves. The model accounts for the pattern and duration of the eye fixations of college students who are interpreting line graphs with three dimensions. Moreover, it provides a theoretical framework for analyzing the processing of more complex graphs, more complex tasks, and other types of audiences (novices vs. experts). This focus on the conceptual aspects of graph interpretation contrasts with the implicit assumption of some researchers and graphic designers that graph comprehension is

Massa to help Alonso see off Hamilton and Button | City A.M. FERRARI’S Felipe Massa has pledged to aid colleague Fernando Alonso’s title challenge in the final three races of the season. Massa’s willingness to effectively perform as the team’s second driver has been in doubt since he was asked to step aside and let Alonso win in July’s German Grand Prix. But the Brazilian is now determined to earn success for the iconic Italian outfit by making it difficult for Alonso’s title rivals. Alonso is joint second in the drivers standings, behind Red Bull’s Mark Webber and level with his team-mate Sebastian Vettel. English pair Lewis Hamilton and Jenson Button are fourth and fifth. Massa, meanwhile, has endured disappointment in his last two races, finishing eighth in Singapore and then crashing out early on in Japan, and is desperate to rediscover form this weekend in Korea. “I would like to arrive back in Sao Paolo for my home race with a good result in my pocket,” he added.

Did You Know? Impairing effects can vary based on the strength of THC, personal tolerance, and mode of use. Edibles affect people differently than inhaling. While both result in a high, the effects of edibles are delayed. Users may consume more than intended before feeling any effects, possibly resulting in higher levels of impairment. Slowed reaction time Trouble controlling speed Lane weaving Poor decision-making HOW DOES CANNABIS IMPAIR DRIVING? Drugs are found in fatally injured drivers more often than alcohol. Cannabis is the most common drug detected.

Hematology, Blood Chemistry, and Bacteriology of the Free-Ranging Mexican Beaded Lizard (Heloderma horridum) Abstract Blood samples from 16 adult, wild Mexican beaded lizards were obtained for complete blood count and serum biochemical analysis. In addition, both oral and cloacal swab samples were collected and submitted for bacteriologic culture. This is the first report of hematologic and bacterial data from free-living beaded lizards. This information will serve as baseline reference values for future health assessment studies in conservation, captive breeding, and research programs.

Interstitial Laser Coagulation in Benign Prostatic Hyperplasia: A Critical Evaluation after 2 Years of Follow-Up Since 1993 we have prospectively followed a cumulative cohort of males with benign prostatic hyperplasia and symptomatic bladder outlet obstruction who underwent interstitial laser coagulation (ILC) of the prostate. We evaluated the safety and efficacy of ILC with respect to relief of symptoms and bladder outlet obstruction. In addition to the critical evaluation of our clinical results, the perineal and transurethral approaches were investigated as they may make a substantial impact on the overall success rate, including prostate size, number of sticks per prostate volume and type of application. A total of 59 patients were treated with the Nd-YAG laser (mediLas fibertom) between April 1993 and December 1996. At the time of reevaluation, 47 patients had completed a follow-up of up to 24 months. A perineal approach was used in 34%, transurethral in 23%, and a combined approach in 43% of the patients, depending on the preoperative volume of the prostate. 75% were high-risk patients in accordance with the ASA score (ASA III). The efficacy of treatment was assessed 6, 12, 24 and 52 weeks postoperatively in accordance with the International Prostate Symptom Score (IPS/quality of life), cystomanometric studies, peak urinary flow rate, residual volume and volume reduction of the prostate. Reduction of prostatic volume and sticks used per prostate volume were correlated to the overall success rate. A significant improvement in all voiding parameters (flow rate, residual volume), including the symptom score, was observed. Pdet decreased from an average of 90 cm H2O preoperatively to 42 cm H2O postoperatively after 24 weeks and the mean reduction in prostate volume was 14 cm3. Interestingly, it was noted that the overall success rate was not size-related. A distinct positive correlation was found in the number of sticks performed and the improvement in objective and subjective parameters. In view of the low morbidity outcome, especially in high-risk patients, we proclaim Nd-YAG interstitial laser coagulation of the prostate to be an effective and safe alternative method of treatment for symptomatic benign prostatic hyperplasia.

Since March is considered National March Into Literacy Month, it is a great time to consider the issue and what we can do to help make improvements. According to the National Coalition for Literacy (NCL), roughly 93 million adults in the U.S. have basic and below-basic literacy skills. Further, around 40 million adults in the country only read or write at or below the 5th Grade level, while another 21 million Americans are not able to read at all. Three Fairfax County Public Schools (FCPS) students are among eight winners of the 2011-12 Student Playwrights Project 10-Minute Play Competition, sponsored by Arena Stage. Fairfax County Public Schools (FCPS) will offer a number of summer programs to provide academic and enrichment opportunities for students in all grades. Visit http://www.fcps.edu/is/summer/index.shtml for general information or the websites listed below for individual programs. Students should register for programs based on the grade they are in during the 2011-12 school year.

Africas Pasts and Africas Historians There are many ways of approaching the past. What is called "history" in European, American, or African universities is only one of them. As V.Y. Mudimbe and Bogumil Jewsiewicki observe, "Africans tell, sing, produce (through dance, recitation, marionette puppets), sculpt, and paint their history". One should not make the division between academic history and other varieties too sharp: the mission graduates of the 1920s or 1930s who wrote down "their" oral traditions were simultaneously mediating between genres, acting within local idioms that phrased political arguments in terms of collective memory, and redefining the source material for academic history. And scholars from Blyden to Cheikh Anta Diop were engaging western philosophical discourse, taking it seriously while refusing to accept basic premises; they confronted the idea of a history of "western civilization" with an "Africa" of mature civilizations and vital influence on other parts of the world.1 Mamadou Diouf's article, which is published in this issue, brings out the rejection of "history" by intellectuals like Ashis Nandy, Ousmanne Sembene, and Archie Mafeje, a rejection of a history that confines the zigzags of time into linear pathways, privileges states over all other forms of human connection, and tells a story of "progress" inevitably leaving Africans or Asians on the side, lacking some crucial characteristic necessary to attain what is otherwise "universal."

Therapeutic approach guided by genetic alteration: Use of MTOR inhibitor in renal medullary carcinoma with loss of PTEN expression Renal Medullary Cancer (RMC) is a rare and aggressive type of renal cell cancer that presents predominantly in patients with sickle cell hemoglobinopathies, and is typically metastatic at the time of presentation. Although platinum based chemotherapeutic regimens have recently emerged as the best option for producing a clinically significant response as reported in various case series, the response is far from satisfactory, as most RMC patients still succumb to their disease within a year of diagnosis. There is currently no standard of care for treatment of this disease. We report, to our knowledge, the first case of RMC where in molecular characterization of the tumor was used to guide therapy. In our patient, molecular analysis identified a decreased expression of Ribonucleotide Reductase M1(RRM1) and phosphatase and tensin homolog (PTEN). Based on these results of PTEN deficiency, we started our patient on everolimus (an MTOR inhibitor) maintenance after treating him with an induction chemotherapy regimen of Paclitaxel-Cisplatin-Gemcitabine (PCG). His tumor responded to induction therapy and he went into complete remission and remained in remission for 7 months. He is now alive about 14 months from his diagnosis and is asymptomatic with minimal disease. The rarity of RMC makes it very difficult to do any meaningful clinical trials in this group of patients. The overall prognosis for RMC remains very poor and knowledge about driver mutations may help in guiding therapy to improve survival in this select group of patients, where there is dearth of available therapies. Introduction Renal Medullary Carcinoma (RMC) is a rare tumor found predominantly in young patients with Sickle Cell Trait (SCT) or Sickle Cell Disease (SCD). 1,2 The disease is highly aggressive, typically presenting with widespread metastases at the time of diagnosis. 3 The disease is predominant in males (M:F D 2:1) with a median age of 26 years and the tumors commonly occur in the right kidney. 4 The pathogenesis of this disease is unknown but in some cases inactivation of SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 (SMARCB1) protein has been observed suggesting its role in the development of this disease. 4,5 Historically, patients would succumb to this disease within weeks to months with a median of <12 months. 1,6 Because of the rarity of the tumor and its vicious nature, clinical trials are difficult to perform. There are some case reports and case series 1 that hint toward the benefit of using cisplatin-based chemotherapy regimen in RMC but almost all of them relapsed quickly after treatment. The overall prognosis of this disease remains poor with no cure. We present here what is, to our knowledge, the first case in which molecular analysis of the tumor was used to direct maintenance therapeutic approach post cisplatin-based chemotherapy in a patient with metastatic RMC. Case Report A 27 year-old African-American male with SCT presented with hematuria and left flank pain in June 2013. Computed tomography (CT) of the abdomen and pelvis revealed a poorly defined mass at the midpole of the left kidney, 4.8 £ 4.4 cm in size, with minimal hydronephrosis (Fig. 1). CT guided biopsy of the mass was performed at an outside institution. The pathology came back as RMC. The patient underwent left radical nephrectomy without any complications in an outside institution. The surgical specimen contained highly atypical cells arranged in nests, sheets, and adenoid cystic patterns, with intratumoral neutrophilic infiltration and peritumoral inflammation and fibrosis (Fig. 1). Staining was positive for thrombomodulin, EMA, CAM5.2, PAX-8, high molecular weight keratin, and E-Cadherin; and negative for PAX-2, uroplakin, p63, and Mucicarmine. The tumor was 6.5 cm in its longest dimension, and was pathologically staged as pT3a pN1 Mx. He had not had a staging CT chest prior to his surgery. Five weeks post his surgery, in July 2013; he was referred to our institution for further management. At that time, he had noticed recurrence of his flank pain. CT of the chest, abdomen, and pelvis revealed mediastinal, hilar, and gastrohepatic ligament lymphadenopathy with right pulmonary nodules suspicious for metastatic disease (Fig. 2). Bone scan had shown suspicious lesion in the left 8th rib, which was thought to be a metastatic site. He was unable to start chemotherapy with cisplatin based regimen because of renal insufficiency and was instead started on carboplatin (AUC 6) and Paclitaxel (175 mg/m 2 ) Q3 weekly with denosumab (120 mg) Q6 weekly (per patients' request to decrease frequent hospital visits) with plans to switch to cisplatin based regimen when renal function improves. We switched him to PCG (paclitaxel 80 mg/m2, cisplatin 35mg/m2 and gemcitabine 1000 mg/m2 on days 1 and 8 every 21 d), when his renal function stabilized with GFR >50. He tolerated the chemotherapy well except for myelosuppression needing dose reduction and addition of neulasta from his C#3 onwards. He got a total of 6 cycles with platinum based chemotherapy, out of which 5 were PCG. CT of chest and abdomen after cycle 2 of PCG revealed marked decrease mediastinal, hilar, and gastrohepatic ligament lymphadenopathy with complete resolution (CR) of lung nodules and lymph nodes post completion of cycle 4 of PCG chemotherapy (Fig. 2). Given the rarity of the tumor and no standard of care for treatment, his surgical specimen was sent to Caris Life Sciences for molecular profiling and next generation sequencing in the hopes of finding a targetable mutation. Tumor immunohistochemistry revealed below threshold expression of Ribonucleotide Reductase M1 (RRM1) expression (Figs. 3A), near complete loss of phosphatase and tensin homolog (PTEN) expression (Fig3 B) and 50% c-MET expression (positive but CISH was negative for cMET amplification); as shown in Figure 4B, loss of PTEN gene function removes the negative feedback mechanism exerted on PI3kinanse, thereby increasing downstream signaling and increased un-inhibited cell growth. The tumor biomarker profile indicated potential benefit of gemcitabine, nabpaclitaxel, and doxorubicin. Since most of the literature showed that patients had early relapse of their disease post chemotherapy, we wanted to explore maintenance therapy for his disease to prevent early relapse. 1,7 His molecular analyses had shown low level of PTEN expression by IHC(5%), frequently correlated with PTEN gene deletion and functional loss. We opted to give him maintenance everolimus (mTOR-mammalian target of rapamycin, inhibitor) 10 mg/daily therapy, starting beginning of December 2013. Figure 4C demonstrates potential targets for therapy in patients with PTEN gene loss of function, including an MTOR inhibitor. His follow up bone scans were stable and upon repeat review, the rib lesion was thought to be a bony island of uncertain significance. He tolerated everolimus well, follow up CT chest and abdomen showed maintenance of CR until the end of June 2014, when his imaging showed a significant increase in his mediastinal and hilar and aortocaval lymph nodes. However, all his nodes were less than 2 cm and he was clinically asymptomatic. The repeat biopsy from his aortocaval lymph node was positive for recurrence of his disease. He was switched to gemcitabine single agent therapy in July 2014 and continues to tolerate it well. He is presently alive and almost 14 months from his diagnosis. We had wanted a repeat biopsy for next generation sequencing but due to unavoidable circumstances this was not possible in spite of disease progression. Discussion The patient in this study presented with RMC that was widely metastatic shortly after the time of presentation. We had done molecular profiling to determine possible targets with intent to treat. He was found to have near absence of PTEN expression by immunohistochemistry and we used this information to guide www.taylorandfrancis.com 29 Cancer Biology & Therapy his maintenance therapy (Fig. 4C). He tolerated this therapy well with no significant side-effect and was in clinical and radiological remission for 7 months since stopping of his induction chemotherapy and is now alive about 14 months since his diagnosis. Our case is one of the very few cases with advanced RMC who achieved CR from his initial chemotherapy. To the best of our knowledge, this is the first case to show that profiling and subsequent targeted therapy had therapeutic benefit in RMC and should be considered in these patients. Our patient is alive; about 14 months post his diagnosis. RMC was first described in 1995 by Davis et al as "the seventh sickle cell nephropathy." Since that initial publication, the literature has become populated with case reports and periodic reviews of the literature, which form the principle body of knowledge available on this tumor. The disease is usually widely metastatic at the time of diagnosis. The tumor microscopically presents with islands of reticular growth, with adenoid cysticlike spaces, solid sheets, and anastomosing tubules. 8 The cancer cells are highly atypical, with a rhabdoid or plasmacytoid shape as well as dark cytoplasm, clear nuclei, and prominent nucleoli. 9 Prominent fibrosis and inflammation are characteristic. Historically, the prognosis of this tumor has been dismal, with patients surviving approximately weeks to months. We searched for all English-language case reports for adult patients (>18 y of age; we have included one case of 17 y old who had an amplification of ABL-tyrosine kinase) with RMC that was metastatic at or shortly after the time of diagnosis, published since 2003, and gathered available data on these cases pertaining to chemotherapy protocols and survival since the time of the diagnosis ( Table 1). We only included patients from the case reports or case series who had data for chemotherapy use, survival post diagnosis and or response to therapy. Our search returned 34 unique cases published during that time, out of which only 17 cases met our above mentioned criteria for inclusion in the Table 1. 1, The median survival calculated for all included patients in the Table 1 was 10.75 months with an average of 10.9 months. We observed that the majority of patients who had any meaningful response (complete or partial response) were treated with cisplatin based chemotherapy (survival appeared to be more extended with PCG regimen when compared to the rest). Majority of these patients had right sided kidney RMC. The observation of positive response to cisplatin appears to be consistent with the previously published notion that renal medullary carcinoma may respond to platinumbased agents. 18,10 While these regimens have improved survival compared to the original studies, they still universally fail to produce a durable response or cure. The difficulty in achieving durable responses for this tumor may reflect our poor understanding of its pathophysiology. A report by Swartz et al. 19 has shown a strong expression of Vascular Endothelial Growth Factor (VEGF), and Hypoxia Inducible Factor (HIF) in RMC. 19 These findings formed the basis for a pathophysiologic theory, which proposes that the low oxygen tension in the renal medulla, especially in patients with sickle cell hemoglobinopathies, promotes the expression of HIF. Under normal circumstances, HIF leads to upregulation of p53, ultimately causing apoptosis. In the setting of deficient or absent p53, however, HIF instead leads to VEGF expression, which may help tumor vascularization and growth. In a separate report, Yang et al. suggested presence of hypoxia related and unrelated genetic abnormalities in significant number of cases, and hinted their role in the pathogenesis of RMC. 20 Recent studies looking into the cytogenetic abnormalities of RMC have failed to identify any consistent abnormalities across cases, and have also brought into question the aforementioned theory by Swartz on the grounds of cases in caucasian patients without hemoglobinopathies. 21,22 Gao et al., had recently shown involvement of genes in signaling pathways such as phosphatidylinositol 3-kinasePIK3C2G), cell cycle control (CDC25D and CDC73), and histone synthesis (HIST2H3D) based on molecular analysis of 11 patient specimens in a retrospective fashion. Although mechanism of pathogenesis still remains largely elusive, these alterations may become probable targets for therapy in the future. 23 It is both the lack of complete understanding of the disease and rarity of desirable response to treatment that led us to our treatment approach. The fact that RMC is probably sensitive to platinum based chemotherapy, coupled with the low expression/ absence of RRM1 in his tumor; may have contributed to the complete remission of our patient's disease when treated with PCG chemotherapy regimen. Whether the low RRM1 expression facilitated a strong response to gemcitabine component in PCG is difficult to determine. The decision to attempt the use of everolimus for maintenance therapy, however, was entirely guided and deemed feasible based on the information regarding loss of PTEN expression gained from IHC profiling of the tumor. It is well known that PTEN deletion causes upregulation of PI3K-AKT pathway which plays a significant role in proliferation and progression of cancer and since mTOR is a downstream signal for the PI3K-AKT pathway, blocking this mTOR signal may cause decrease in tumor proliferation. 24 There is literature supporting PTEN deletion being a marker for predicting response to everolimus in other cancer types such as prostate cancer and that was our rationale for using everolimus in this patient. 24 Conclusion We acknowledge that molecular profiling with next generation sequencing is not feasible, nor is it necessary for every patient with a malignant tumor. However, for these rare tumor types, especially those in which treatment guidelines are not well established, comprehensive molecular characterization may provide an invaluable insight for choosing a more personalized therapeutic approach. The information from molecular profiling can be used to guide maintenance therapy with targeted agents, which are better tolerated than chemotherapy and may provide a road to prolonged survival in patients with this deadly disease in this current era of personalized medicine. Disclosure of Potential Conflicts of Interest No potential conflicts of interest were disclosed. Ethical Statement The authors complied with patient confidentiality. An informed consent was obtained by the patient prior to preparing this manuscript. Authors Contribution JSL and SMR-manuscript preparation and both contributed equally to be co-first authors; ZG and NES-manuscript preparation, critical review of the article; SLH, MK and JJD provided critical review of the article; MJ-manuscript preparation, critical review/revision of the article and patient care. Abbreviations: RN-radical nephrectomy; GC-gemcitabine and cisplatin; GCa-gemcitabine and carboplatin; PCG-paclitaxel, cisplatin, gemcitabine: PCaGpaclitaxel, carboplatin, gemcitabine; PCa-carboplatin, paclitaxel; MVAC-methotrexate, vinblastine, doxorubicin, cisplatin; AG-adriamycin, gemcitabine; EDCV-Etoposide, Doxorubicin, Cyclophospamide, Vincristine; GD-gemcitabine, docetaxel; ICE-ifosfamide, carboplatin, etoposide; TC-thalidomide, capecitabine; PD-progressive disease; PR-partial response; SD-stable disease; NA-not available; f/u,followed by *-post salvage. He had topoII increased expression by immunohistochemistry; **-no response to everolimus and it was used as last resort for a few weeks. Reason to start this unclear but she died very shortly after that; ***-this is the only patient we have included who is <18 years because her tumor had amplification of ABL-tyrosine kinase.

CMV in the gut: a critical review of CMV detection in the immunocompetent host with colitis As scientific techniques for the detection of cytomegalovirus (CMV) improve, we are able to detect small amounts of CMV in the mucosal wall. As clinicians, we are unsure how to interpret the results of this novel test. There is controversy in the literature as to the significance of the detection of CMV in the gut. Whilst the importance of CMV and reactivation of the virus is clear in those patients such as allograft recipients with established immune compromise, the role is less clear in patients with less damaged immune systems. We explore whether the detection of CMV in such cases influences outcome and how it should be optimally managed. We discuss the optimal management of such cases, according to current guidelines, with a review of the literature. Introduction In the apparently immunocompetent patient presenting with bloody diarrhoea for the first time, there is often diagnostic uncertainty as to whether the cause of colitis is likely to be infectious or inflammatory. Risk factors for an infectious aetiology include undiagnosed immune compromise , travel, exposure to antibiotics or hospital, and unprotected sexual intercourse. Even when an infectious colitis is diagnosed, this does not exclude the possibility of a subsequent diagnosis of an inflammatory bowel disease, which may present initially as an infective colitis. In a first episode of severe colitis which fails to respond to steroids or other immunosuppressive therapy, guidelines suggest that we should look for cytomegalovirus (CMV) at sigmoidoscopy ; however it is acknowledged that the subsequent detection of CMV can be difficult to interpret. The diagnosis of CMV colitis in the immunocompetent host is rare, but needs to be considered, alongside the possibility of undiagnosed immunocompromise, in the patient that fails to respond to maximal immunosuppressive therapy. Case reports and case series of immunocompetent patients with CMV colitis have been published; however, the patients included often had comorbidities or pregnancy, which could affect their immune status. Although CMV may be detected at diagnosis in an immunocompetent patient with ulcerative colitis, it is more commonly diagnosed following immunosuppressive treatment for colitis. The literature regarding the importance of the positive CMV result on biopsy is reviewed here with respect to the following questions: Is CMV ever the sole cause of colitis in an immunocompetent host? CMV is a major cause of morbidity in immunosuppressed patients, causing significant disease in transplant patients and, prior to the introduction of highly active antiretroviral therapy (HAART), in HIV. In such patients, end-organ involvement following viraemic spread of CMV may lead to damage to a single organ, as seen in, for example, colitis, retinitis or severe pneumonitis. CMV is also known as human herpesvirus 5 and, like other herpesviruses, it causes a primary infection followed by the establishment of latency, a dormant infection in which only a few genes are expressed. Recurrent disease can occur if the virus reactivates due to perturbations in immunity, e.g. as seen with age or immunosuppressive drugs. CMV is common, with a seroprevalence (CMV IgG-positive) of 40-100 % in adults, increasing with age. Primary infection is often asymptomatic in the immunocompetent host but can cause a mild febrile illness and an infectious mononucleosis syndrome. However, single-organ pathology, such as hepatitis, retinitis or colitis, occurs rarely following primary infection or reactivation in an immunocompetent host. CMV colitis in the immunocompetent patient is uncommon, though it has been described as presenting with a syndrome incorporating symptoms of colitis (e.g. abdominal pain, fever, diarrhoea, rectal bleeding). Galiatsatos et al. reviewed the literature and found 44 immunocompetent patients with CMV colitis; however, 34 of these patients had comorbidities that would be expected to affect immune function (pregnancy, renal disease, diabetes, malignancy). In that study, age over 55 years was found to be associated with a poor outcome. This small study suggested that the diagnosis in an immunocompetent patient is rare and raises the question of either an alternative diagnosis, such as a new presentation of inflammatory bowel disease (IBD), or a previously undetected immune deficiency (such as HIV). Even mild immunosuppression, as seen in chronic kidney disease, seems to predispose to CMV reactivation and colitis. Therefore, the answer to this question is probably yes, but rarely. How can we diagnose CMV colitis? Although a wide range of diagnostic tests for CMV is available, each has limitations (Table 1). Serology is useful to establish evidence of previous infection (CMV IgG) and IgG avidity can help to estimate the time of primary infection, as the IgM antibody can remain positive for up to a year following primary infection. If CMV colitis is suspected, the bowel is examined endoscopically for evidence of CMV disease. This may be detected as typical findings on histology, such as owl's eye inclusion bodies. This histological appearance is very specific for CMV, has a clear relation to polymerase chain reaction (PCR) detection of CMV in the gut and provides the mainstay of diagnosing CMV end-organ disease post-transplant. However, histology has a low sensitivity, so CMV infection may be missed. Immunohistochemistry (IHC) or simple haematoxylin and eosin (H&E) staining can be used to improve sensitivity if a diagnosis of CMV colitis is considered (Fig. 1). Histology, H&E and IHC stains retain specificity for CMV disease. CMV DNA detection in the blood by PCR has replaced the previous technique of CMV pp65 antigen detection and high levels of CMV DNA in the blood correlate with positive IHC and detection in tissue. However, 15 % of people with end-organ damage causing retinitis were found to have no CMV detectable in their blood in the pre-HAART era, suggesting that viraemia does not always persist until the time of clinical presentation. Modern studies suggest that CMV PCR in stool may, in due course, allow us to detect CMV with a non-invasive test. Unfortunately, currently, the most sensitive test for CMV, CMV biopsy PCR, does not seem to have sufficient specificity to be used alone to make a diagnosis of CMV colitis. CMV viral cell culture is also sensitive but labour-intensive and has, therefore, been replaced by nucleic acid detection. As these tests detect CMV at very low levels, they also detect small amounts of reactivating virus that may not be causing disease. In the immunocompetent host, periodic reactivation of CMV does not necessarily indicate pathology. The significance of such small amounts of CMV in the colon has not been established. Lawlor and Moss found that the majority of published studies detected CMV DNA in the bowel in a significant proportion of patients without detection by H&E or IHC. They suggest that the use of a 'cut-off' DNA level might aid diagnosis where CMV in the colon could be considered important only when present at above a certain level. That level has not yet been established and further work is clearly needed in this area. Does the presence of CMV correlate with disease activity? CMV has been found in approximately a third of samples from patients with severe ulcerative colitis. In contrast, in inactive colitis, the detection of CMV DNA is rare. Although CMV DNA is detected more often in severe colitis, inflamed tissue to further propagate infection (Fig. 2). If CMV was a significant factor in pathology, one would expect those patients with IBD and positive CMV IgG to have a worse outlook than patients with negative CMV IgG, but in a study of 187 patients, the two groups were found to have similar appearances of mucosa at colonoscopy. In addition, the presence of CMV DNA in blood does not seem to predict the deterioration of IBD in prospective studies. Is CMV a precipitant for IBD? Although the detection of CMV has been found to be associated with severe disease in IBD, there is no evidence that this relationship is causative. CMV at presentation of ulcerative colitis or IBD does occur, but is rare. Blood PCR is often positive after 2-3 weeks of steroid treatment but this rarely leads to CMV disease on histology with DNA levels lower than those seen in transplant patients and CMV DNA levels falling as steroids are reduced. In transplant patients, the administration of steroids reduces the CMV viral load required to cause end-organ disease and perhaps this interaction also occurs in the immunocompetent patient. Does the presence of CMV simply reflect the degree of immunosuppression? CMV is detected more often in cases of steroid-refractory disease. It is unclear if such patients would have more CMV because they have more inflammation or because they have received greater doses of immunosuppression in an effort to control their colitis. In some patients, CMV detection occurs prior to the prescription of immunosuppressants, but in IBD, CMV is commonly detected following immunosuppressant therapy. Matsuoka et al. demonstrated a worse prognosis in those with high levels of CMV in blood leading to greater requirements for ciclosporin. In turn, patients treated with ciclosporin also develop more CMV and have a poorer prognosis. Studies in organ transplant recipients have shown that higher levels of immunosuppression lead to Fig. 3 Flow chart of suggested management higher rates of CMV disease. Extrapolated to IBD, this suggests that more CMV may be seen when more immunosuppressants are used. In the case of the detection of CMV DNA in the mucosal wall in the presence of colitis, it is usually unclear whether the CMV is the cause of pathology or an innocent bystander. Can treating CMV improve outcome in patients with colitis? Although guidelines are universal in recommending the treatment of CMV in severe refractory colitis with CMV detected, this guidance is based on a number of small studies with inconclusive outcomes. A randomised controlled study in HIV in the pre-HAART era showed a benefit of treating CMV colitis in acquired immunodeficiency syndrome (AIDS), but no randomised controlled studies have been done in immunocompetent hosts. Those retrospective studies or observational studies performed in immunocompetent hosts are confounded, as treatment with ganciclovir is more often used in patients with more severe disease. It is known that, in the absence of anti-CMV treatment in colitis, reactivated CMV may resolve spontaneously. CMV viraemia post-transplant also frequently resolves without specific anti-CMV treatment. There are insufficient published data to determine if treating CMV impacts upon major outcomes such as colectomy rates and mortality. How should we manage a patient with CMV detected on colonic biopsy? The American College of Gastroenterology (ACG) and the European Crohn's and Colitis Organization (ECCO) recommend treatment of CMV with antivirals only when a patient with severe colitis is failing to respond to immunosuppressive therapy. The ECCO state that, when detected only by PCR, in the absence of supportive histology or immunohistochemistry, the detection of CMV may not always suggest disease. They recommend the discontinuation of immunosuppressive agents only in cases of severe colitis with detection of CMV in the mucosa but do not state clearly in what form the detection may occur. The British Society of Gastroenterology (BSG) refers to the ECCO guidance but consider the situation in greater detail. They describe the situation in which active CMV colitis in a patient on immunosuppression is difficult to distinguish from a flare of IBD. They suggest treatment of CMV and discontinuation of immunomodulators in severe or refractory colitis in which CMV is detected by histology and PCR. Our proposed flow chart (Fig. 3) illustrates the British approach to management. However, these guidelines may be outdated as more recent data suggest that early detection and treatment of CMV may be beneficial. Roblin et al. found that a CMV DNA load above 250 copies/mg in tissue was predictive of resistance to three successive treatment regimens for ulcerative colitis and suggest that it might be prudent to treat those patients found to have high CMV DNA levels before they deteriorate. Summary The current position with respect to the detection of CMV in the bowel mucosa in colitis is unclear. This is particularly important in the patient with severe colitis on immunosuppression and detection of CMV. The risks and benefits of reducing immunosuppression and prescribing treatment for CMV in such a patient are not determined. There is an urgent need for further work in this field. Studies are needed in order to clearly define the significance of a positive CMV in the bowel (or stool) in colitis; to determine if patients with CMV detected by PCR, but not biopsy-proven, would benefit from antiviral treatment; and to determine whether reduction in immunosuppression causes more harm than good in such patients. Conflict of interest The authors declare no conflicts of interest. Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.

British Columbia's Lieutenant-Governor has asked the New Democrats to form government, ending 16 years of Liberal rule after a confidence vote in the legislature. Premier Christy Clark's resignation ends the political uncertainty that has hung over the province since the May 9 election, and will allow the NDP to take power with a potentially fragile arrangement with the third-place Greens. Here's what you need to know about what has happened so far, and what comes next. Story continues below advertisement Related: Judith Guichon, the Lieutenant-Governor who will decide B.C.'s fate Related: New Brunswick holds lessons for B.C. on Speaker traditions Gary Mason: In a strange few weeks for B.C., Clark risked her political past and future Here's what you need to know about the confidence vote and what happens next. The confidence vote Lost confidence The non-confidence motion was proposed as an amendment related to the Throne Speech. It was moved by NDP Leader John Horgan and seconded by Green MLA Sonia Furstenau. It reads: Be it resolved that the motion "We, Her Majesty’s most dutiful and loyal subjects, the Legislative Assembly of British Columbia in Session assembled, beg leave to thank Your Honour for the gracious Speech which Your Honour has addressed to us at the opening of the present Session," be amended by adding the following: "but Her Honour’s present government does not have the confidence of this House." The Opposition New Democrats, armed with a power-sharing agreement with the B.C. Green Party, tabled an amendment to the Throne Speech that declared the legislature does not have confidence in the government. The outcome was not a surprise; the Liberals finished the May 9 election one seat shy of a majority and were short a vote with a member in the Speaker's chair. As the standing vote was called, a hush fell over the packed House. When the roll was called, 44 MLAs – all the Greens and New Democrats – had won the motion. The Liberals, with 42 votes, were easily defeated. A visit with the viceregal Ms. Clark visited Lieutenant-Governor Judith Guichon shortly after the confidence vote. The premier had three options: request an election; advise the Lieutenant-Governor to ask the Opposition New Democrats to form government; or simply resign without offering any advice. Story continues below advertisement Ms. Clark said earlier this week that, if asked, she would tell the Lieutenant-Governor that the current makeup of the legislature means it cannot function – even with the NDP-Green agreement in place. The premier said she would stop short of asking for an election, though her comments fuelled speculation that she was hoping for a campaign. However, Ms. Guichon appeared unconvinced by those warnings, instead inviting NDP Leader John Horgan to form a government. Shortly after the confidence vote, Mr. Horgan said he was eager to get to work. "I'm excited. Now, seven weeks after the election, we can get going on a government that works for the people," he said in an interview while waiting to hear from the Lieutenant-Governor. The Liberals had suggested the current rules that govern the legislature make a bare, one-seat majority unworkable. Immediately after the vote, Liberal Steve Thomson resigned as Speaker, meaning the NDP will need to offer up a member to fill that role. Since the Speaker doesn't vote except in a tie, he or she will be routinely called upon to break votes in the legislature. The rules around committees are even more complicated and could actually leave the NDP-Green alliance with a minority in some instances. Story continues below advertisement However, the New Democrats say they can use the committee system to their advantage, without having to rewrite the rules of the house, to manage the debate and reduce the need for the Speaker to participate in votes. A change in government The NDP have only ever won three elections in B.C. and haven't won since 1996. This time, they intend to govern with the support of the Greens, led by Andrew Weaver, who have agreed to vote with the New Democrats on confidence votes for four years in exchange for commitments on a series of policies. Mr. Horgan is now premier-designate (Ms. Clark and her cabinet remain in place in the meantime) and will spend the coming weeks preparing to take power and deciding on a cabinet. The new premier and his cabinet will likely be sworn in by the end of July. The New Democrats will then draft a budget plus a series of bills that they have promised to introduce as part of their accord with the Greens. As well, they will launch a review of the $8.8-billion Site C dam which could lead to the cancellation of the province's most expensive public infrastructure project in history. However, the NDP and Green pact could face challenges. The two parties will find points of discord on issues that were not included in their agreement; Mr. Weaver sahd his party will handle those issues on a case-by-case basis and his MLAs would not be whipped on votes that aren't matters of confidence. The standings in the legislature, even with the NDP-Green alliance, will still present challenges. The Speaker would be routinely called upon to break ties, which experts have said would require him or her to break with centuries of convention. As well, will be no margin for MLAs to be absent for travel or illness. The Lieutenant-Governor's dilemma The Lieutenant-Governor had been getting advice from her counterparts elsewhere in Canada and throughout the Commonwealth to guide her decision. Some constitutional experts suggested it would be unlikely for Ms. Guichon to trigger a vote so soon after a general election, especially if the New Democrats and Greens have an agreement in place, however fragile. The two examples in Canadian history most often cited have been the King–Byng Affair in the 1920s and the fall of Frank Miller's government in Ontario in 1985; in both instances, the viceregals allowed the Opposition to form government rather than dissolving the legislature. However, neither of those cases involved legislatures where the standings were so tight. In the end, Ms. Guichon appeared to stick with those precedents, giving the New Democrats to attempt to govern before considering a new election. Philippe Lagassé, an associate professor and constitutional scholar at Carleton University, said it was the right call. "She can't look at the legislature and start making predictions about how it may or may not work," he said before the Lieutenant-Governor revealed her decision. "Ultimately, she is going to have to say, 'Is there a viable majority that will hold confidence or not?' and to my mind, that is really the best course of action for her." MORE FROM THE GLOBE AND MAIL: How a deal with the Greens slipped away from Christy Clark’s BC Liberals The BC Liberals came close to forming an alliance with the Green Party that would have salvaged Premier Christy Clark’s government. But in the end, it was an arrangement many Greens simply could not stomach BC enters the fragile world of minority governments As B.C. prepares for life under a minority government, the province can learn lessons from legislatures that have been there

1-Benzyl-3-methylimidazolium chloride 0.25-hydrate The asymmetric unit of the title compound, C11H13N2 +Cl−, contains two independent ion pairs and and half a solvent water molecule (m site symmetry for the O atom). The imidazole ring is oriented at dihedral angles of 66.61 and 89.17° with respect to the aromatic ring in the two cations. In the crystal, OH⋯(O,Cl) hydrogen bonds and stacking interactions between the imidazole ring of one molecule and the aromatic ring of another link the molecules. Comment Ionic liquids (ILs) are a class of compounds composed of organic cations and organic or inorganic anions and have attracted significant attention due to their beneficial properties and their low impact on environment. Many investigations have been dedicated to their use as new media for synthetic chemistry (), catalysts for organic synthesis (;), extractants for separation science () and electrolytes for electrochemistry () as well as other areas due to their peculiar physical properties such as a wide liquid range, non-volatility, chemical stability and large windows of electrochemistry. Because it is technically difficult for ionic liquid to grow single crystal and to select suitable sample for single crystal x-ray diffraction at low temperature, so far a few crystal structures have been determined by x-ray diffraction (). We have synthesized a numer of ionic liquids. The title compound is one of the products, and we report herein its crystal structure. In the molecule of the title compound ( Fig. 1) the bond lengths () and angles are within normal ranges. The imidazole ring is oriented with respect to the aromatic ring at a dihedral angle of 89.17 ° and 66.61 °, respectively. The hydrogen bond are formed between one Cl and H 2 O (Table 1). In the crystal structure, - packing between the imidazole ring of one molecule and the aromatic ring of the other link the molecules. The packing diagram of the O-HCl bonds is shown in Fig. 2. Experimental For the preparation of the title compound, N-methylimidazole (8.2 g, 0.1 mol) was dissolved in dry acetonitrile (30 ml). Refinement H atoms of water were located in a different Fourier map and the atomic coordinates allowed to refine freely. Other H atoms were positioned geometrically and refined as riding (C-H = 0.93-0.96) and allowed to ride on their parent atoms, with U iso (H) =1.2U eq (parent) or 1.5U eq (parent). supplementary materials sup-2 Figures Fig. 1. The molecular structure of the title molecule, with the atom-numbering scheme. Displacement ellipsoids are drawn at the 30% probability level.

Outcomes of bone marrow mononuclear cell transplantation combined with interventional education for autism spectrum disorder Abstract The aim of this study was to evaluate the safety and efficacy of autologous bone marrow mononuclear cell transplantation combined with educational intervention for children with autism spectrum disorder. An openlabel clinical trial was performed from July 2017 to August 2019 at Vinmec International Hospital, Hanoi, Vietnam. Thirty children who fulfilled the autism criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition, and had Childhood Autism Rating Scale (CARS) scores >37 were selected. Bone marrow was harvested by anterior iliac crest puncture under general anesthesia. The volume collected was as follows: 8 mL/kg for patients under 10 kg (80mL+7 mL) for patients above 10 kg. Mononuclear cells were isolated with a Ficoll gradient and then infused intrathecally. The same procedure was repeated 6 months later. After the first transplantation, all patients underwent 8weeks of educational intervention based on the Early Start Denver Model. There were no severe adverse events associated with transplantation. The severity of autism spectrum disorder (ASD) was significantly reduced, with the median CARS score decreasing from 50 (range 4055.5) to 46.5 (range 33.553.5) (P<.05). Adaptive capacity increased, with the median Vineland Adaptive Behavior Scales score rising from 53.5 to 60.5. Social communication, language, and daily skills improved markedly within 18months after transplantation. Conversely, repetitive behaviors and hyperactivity decreased remarkably. Autologous bone marrow mononuclear cell transplantation in combination with behavioral intervention was safe and well tolerated in children with ASD (Trial registration: ClinicalTrials.gov identifier: NCT03225651). | INTRODUCTION Autism spectrum disorder (ASD) is a complex spectrum of disorders characterized by two typical abnormalities: (a) deficits of social communication and interaction; (b) the presence of restricted interests as well as repetitive and stereotypic verbal and nonverbal behaviors. 1,2 Comorbidities including sleep disorders, seizures, and gastrointestinal difficulties are very common in children with ASD. 3 The prevalence of identified ASD is increasing. 4 In 2016, the overall ASD prevalence among children aged 4 years was 15.6 per 1000 (1/64), 5 and the incidence was 18.5 per 1000 (1/54) in 8-year-old children according to Early Autism and Developmental Disabilities Monitoring Network sites. 6 The etiology of ASD is still not well understood. However, many associated factors including genetic mutations, immune dysregulation, hypoperfusion of some parts of the brain, exposure to maternal antibodies during pregnancy, and weak functional connectivity across brain regions are suggested to contribute to the development of ASD. Multiple approaches including behavioral therapy, occupational therapy, speech therapy, and medications are required in the management of ASD to ameliorate autistic symptoms. Educational and behavioral interventions have been recognized as crucial for the management of ASD in children. 12 The evidence indicates that young children with ASD benefit from interventions that focus on improving social interaction, communication, and challenging behaviors. 13,14 Unfortunately, many children who receive those treatments remain significantly impaired. 15 In search of better outcomes in the management of ASD, alternative and complementary treatments are being investigated. Recent reports have suggested that stem cell transplantation result in improvements in several different neurological conditions. The suggested mechanisms of action of mesenchymal stem cells (MSCs) on the nervous system include neuroprotection, neurogenesis, and synaptogenesis. Stem cell applications were also assessed in animals using the inbred BTBR T+tf/J (BTBR) mouse strain, 22 which has autistic-like symptoms, to explore the potential of stem cells in the management of ASD. In BTBR mice, Segal-Gavish et al showed that transplantation of MSCs resulted in a reduction in stereotypic behaviors, a decrease in cognitive rigidity, and an improvement in social behavior. Moreover, it has been shown that brain-derived neurotrophic factor protein levels as well as neurogenesis increased in the hippocampus following stem cell treatment. 23 Similarly, Perets et al demonstrated that brainderived neurotrophic factors secreted by transplanted MSCs were a key factor in the observed reductions in stereotypic behavior and in the improvement of cognitive flexibility in the BTBR model. 24 Ha et al revealed that transplanted human adipose-derived stem cells improved repetitive behaviors, social interaction, and anxiety in valproic acid-induced ASD model mice. 25 Based on results obtained from animal research, stem cell transplantations have been conducted for children with ASD at several centers. 26 Two distinct approaches have been explored thus far in the application of cell therapy products for the treatment of ASD: culture expanded and nonexpanded. In an early example of non-cultureexpanded cell therapy, Sharma et al described the use of autologous bone marrow mononuclear cell (BMMNC) transplantation infused via intrathecal route in 32 children with ASD. The procedure was reported as being safe with minor adverse events encountered, such as nausea, vomiting, and pain at the site of injection. Improvements were noticed in different aspects, including social relationships and reciprocity, speech and language patterns, and brain metabolism. 27 A further example of concentrated, but not expanded, cell therapy described the use of autologous BMMNC transplantation via the intrathecal route in 10 children with ASD. The results revealed that the maximal treatment effect was observed within the first 12 months with, again, with no safety concerns. 28 A combined culture-expanded/nonexpanded cell therapy study reported on the outcomes of allogenic cord blood mononuclear cell (CBMNC) transplantation vs CBMNC combined with umbilical cord MSC transplantation for children with ASD. 29 In this trial, four stem cell infusions were carried out via intravenous and intrathecal routes. No severe adverse events after cell transplantation were Lessons learned The combination of cell therapy and educational intervention may improve clinical manifestations such as social communication, language, and daily skills in children with ASD. Significance statement The combination of cell therapy and educational intervention may improve clinical manifestations, such as social communication, language, and daily skills, in children with autism spectrum disorder. However, additional studies with control groups should be performed in the future to obtain a more comprehensive and accurate conclusion. described for either group. Improved outcomes were noted in the combination group compared with the CBMNC-alone group. More recently, Dawson et al reported autologous stored cord blood infusion through intravenous route in 25 children with ASD, which resulted in significant improvements in behavior at 6 months after infusion; these improvements were sustained at 12 months. 30 That study was followed by the first randomized, double-blind, placebo-controlled clinical trial comparing outcomes of autologous cord blood infusion vs placebo for children with ASD. As in all previous studies, autologous intravenous cord blood infusion had no serious adverse events and trended toward improvement, especially in socialization, but clinical outcomes were not significantly different between the two groups. 31 In summary, a number of clinical trials have been performed thus far, exploring the application of cell therapy for the treatment of ASD. Although the trials have been broadly consistent in outcome reporting, disparities remain around cell sources, processing, dosage, and delivery route. The aim of this study was to investigate the safety and clinical outcomes of high-dosage BMMNC transplantation combined with educational intervention for children with ASD. | Exclusion criteria The exclusion criteria were epilepsy; hydrocephalus with ventricular drain; coagulation disorders; allergy to anesthetic agents; severe health conditions such as cancer or heart, lung, liver, or kidney failure; and active infections. Patients with Rett syndrome or fragile X syndrome were also excluded from this study. is classified into three levels: level 1 ("Requiring support"), level 2 ("Requiring substantial support"), and level 3 ("Requiring very substantial support"). 33 CARS consists of 14 domains assessing behaviors associated with ASD, with a 15th domain rating general impressions of ASD. 34 VABS-II is a standardized measure 35 that yields an overall score and subscale standard scores in the four different domains: socialization, communication, daily living skills, and motor skills. CGI is a rating scale that measures symptom severity and treatment response. 36 The severity is categorized into seven levels: (a) not present (no ASD), (b) barely evident ASD symptoms, (c) mild ASD symptoms, (d) moderate ASD symptoms, (e) moderately severe ASD symptoms, (f) severe ASD symptoms, or (g) very severe ASD symptoms. The response of each patient is also divided into seven levels: level 1, very much improved; level 2, much improved; level 3, minimally improved; level 4, no change; level 5, minimally worse; level 6, much worse; and level 7, very much worse. In addition, main indicators including social interaction, eye contact, expressive language, abnormal behaviors, sensory abnormalities, eating and sleeping difficulties, daily skills, and learning capacity before and after transplantation were collected to examine the effects of stem cell therapy in combination with behavioral intervention. | Study design A brief video was recorded at baseline and then at 6, 12, and 18 months after the first transplantation. A questionnaire was completed by patients' caregivers 6 and 18 months after the first transplantation to obtain their assessment and satisfaction. A psychologist kept contact with patients' caregivers via telephone throughout the follow-up period to record any unexpected events during the study. | Laboratory and imaging diagnostics Routine hematologic and biochemistry examinations were performed at baseline and 6 months later. Diagnostic imaging examinations, including brain magnetic resonance imaging (MRI) and electroencephalography (EEG), were carried out in all patients at baseline to rule out epilepsy and brain malformations. Positron emission tomography-computed tomography (PET-CT) was carried out before stem cell transplantation and retaken 12 months after the first transplantation to monitor changes of brain metabolism. On post hoc analysis, decreased and increased fluorodeoxyglucose (FDG) metabolism regions in autistic children were evaluated based on a normal distribution curve. If the measured value is lower than one SD from the median value of the standardized uptake value, it is considered to be hypometabolic, whereas a measured value greater than a SD from the median value is considered to be an increase in metabolism. 37 In evaluation of PET-CT images, dark blue brain areas were defined as severely reduced FDG metabolic rate, and light blue brain areas were assessed as moderate metabolism. Green was considered to have mild metabolic reduction. The yellow-orange brain region was assessed to exhibit increased FDG metabolism. 38 Human Build 37 (GRCh37) reference genome. 39 The Genome Analysis Toolkit 40 and SnpEff (an open-source tool that annotates variants and predicts their effects on genes) 41 | Genetic testing We detected and validated a de novo CNV in the SHANK3 gene from a female proband (proband A27) and 23 different variants in 22 genes from eight probands ( SHANK3, CHD8, ANK2, and GIGYF2 belong to the genes with the strongest evidence of relevance to ASD. | Brain MRI and EEG No abnormalities on MRI or EEG were observed in any of the patients. | FDG changes after BMMNC transplantation At baseline, manifestation of hypometabolism was found at seven major brain regions: hippocampus, anterior cingulate gyrus, posterior cingulate gyrus, parietal lobe, frontal lobe, temporal lobe, and central sulcus. After the stem cell transplant, 29 children underwent a second PET-CT scan (the parent refused the second PET-CT in one patient). Improvement in metabolism was observed in some brain regions where severe hypometabolism was noticed before BMMNC transplantation such as parietal lobe, frontal lobe, and anterior cingulate gyrus. However, these changes were not statistically significant (Table 6). | Adverse events None of the patients had any severe adverse events during bone marrow aspiration, stem cell infusion, or following transplantation. The procedure was reported as being safe with minor adverse events encountered. There were no procedure-related major adverse events. Among 96 adverse events that occurred during study period, 46 (48%) mild and moderate adverse events were recorded that may or may not be related to BMMNC transplantation with symptoms including pain, vomiting, and mild fever. All those adverse events were easily managed through appropriate medication (Table 7). | Clinical outcomes After BMMNC transplantation, the severity of ASD decreased remarkably. The median CARS scores decreased from 50 (range 40-55.5) points at baseline to 46.5 (range 33.5-53.5) after 18 months. Patient-specific CARS score analysis for each patient is presented in Figure 1. The result of the mixed-effects analysis suggests that each visit was associated with a decrease of 1.6 in the CARS score and that this change was statistically significant (Table 8). According to the DSM-5 classification, the number of patients at level 3 was reduced from 28 to 18. There were no patients at level 1 before transplantation, but after transplantation, five children were recategorized at this level. After transplantation, improvements were observed in various aspects. Social interaction and eye contact increased remarkably from 37% before the first transplantation to 97% and from 23% to 93%, respectively, after 18 months. Expressive language increased from 47% before transplantation to 67% after 6 months, 87% after 12 months, and 93% after 18 months (Table 2). Abnormal behaviors also decreased after transplantation. The children with repetitive behavior decreased from 93% to 87% (Table 3). Sensory abnormalities and sleeping difficulties progressively improved after transplantation, from 97% to 73% and from 53% to 43%, respectively ( | DISCUSSION In this study, bone marrow was collected from anterior iliac crests and not from posterior crests as in other reports. 17,27 This change facilitated anesthesia and reduced risks related to the prone position of patients. The dosage of transplanted mononuclear cells and CD34 + cells in our study was higher than those in studies reported by Sharma,Dawson,and Chez. 27,30,31 Differences related to the cell transplants compared with other studies are presented in Table 9. Granulocyte-colony stimulating factor (G-CSF) was not used in this study to mobilize stem cells from bone marrow, with high dosage values nevertheless isolated. This implies that G-CSF may not be The incidence of minor adverse events was low and easily managed through medication or spontaneously resolved themselves. Although all participants still belonged to severe level at the baseline after receiving behavioral intervention with a mean duration of 3.5 years, this study showed improvements in various aspects after BMMNC transplantation combined with educational intervention. Positive changes in social communication, eye contact, language, behaviors, and daily skills were observed after BMMNC transplantation. In addition, learning ability also remarkably improved after transplantation. The number of children who could go to school without support increased after transplantation (Table 10). Hyperactivity of children with ASD is a disorder that severely impairs quality of life for the whole family. In our study, the rate of children with hyperactive disorder decreased by 50% at 18 months after stem cell transplantation. Positive changes were found in evaluation measures, including severity and adaptive ability. The number of patients at level 3 (requiring very substantial support) according to DSM-5 decreased from 28 to 18 at 18 months after transplantation. We noticed that the improvements appeared to be influenced by the CARS scores at baseline. Patients with a CARS score ≤49 at baseline showed better improvement than those who had CARS scores >49 points. This would imply that patients with lesser severity had better outcomes after transplantation. Genetic abnormalities have been reported in many ASD studies. In our series, genetic variations were found in eight patients (26%). Two different pathways for administration have been applied in the delivery of cell therapy for ASD: intravenous and intrathecal. Although both delivery routes are safe, there are concerns regarding cells delivered through the intravenous route, as transplantations in animal models have shown that the transplanted cells have difficulties passing through organs such as the spleen, kidney, and intestine. 57 The intrathecal route does not present this concern. The number of BMMNCs transplanted also varies between studies, and the number we have used was higher than those from other groups. There have been suggestions of correlation between transplanted stem cell dosage and the extent of subsequent clinical improvement. 58 Although we did not perform an escalating dosage study, we noted that a high dosage of stem cells may be used to obtain satisfactory outcomes in children with ASD. Furthermore, we performed two transplantations instead of one transplantation as described elsewhere. 27,30,31 Multiple transplantations have resulted in positive outcomes in both ASD 29 and spinal cord injury. 59 Moreover, the benefits of repeated transplantations vs single transplantation have been identified in animal myocardium infarction models. 60 In our study, all children received 18 months of follow-up after the first transplantation. This follow-up duration exceeds those in other recent reports. 27, We noticed that the longer the follow-up duration was, the lower the severity of ASD (CARS score reduction) and the better the children's adaptive functioning (VABS score increase). Meanwhile, there was no case in which the results were not improved or even worse compared with baseline, implying that the treatments have a sustainable long-term effect. It is likely that further extended follow-up times will be required to fully assess the responses of children with ASD over time after stem cell transplantation. Herein, we have demonstrated the safety and feasibility of BMMNC transplantation for the treatment of ASD. The lack of a control group who received either stem cell transplantation or educational intervention only is a limitation of this study. However, our study results provide initial evidence to justify conducting a randomized clinical trial with control groups in the future. | CONCLUSION We conclude that autologous BMMNC transplantation is safe. The DATA AVAILABILITY STATEMENT All data generated or analyzed during this study are included in this published article and its supplementary information files.

Chronic colitis upregulates microRNAs suppressing brain-derived neurotrophic factor in the adult heart Ulcerative colitis and Crohns disease are classified as chronic inflammatory bowel diseases (IBD) with known extraintestinal manifestations. The interplay between heart and gut in IBD has previously been noted, but the mechanisms remain elusive. Our objective was to identify microRNAs mediating molecular remodeling and resulting cardiac impairment in a rat model of colitis. To induce chronic colitis, dextran sodium sulfate (DSS) was given to adult rats for 5 days followed by 9 days with normal drinking water for 4 cycles over 8 weeks. Echocardiography was performed to evaluate heart function. DSS-induced colitis led to a significant decrease in ejection fraction, increased left ventricular mass and size, and elevated B-type natriuretic protein. MicroRNA profiling showed a total of 56 miRNAs significantly increased in the heart by colitis, 8 of which are predicted to target brain-derived neurotrophic factor (BDNF). RT-qPCR validated the increases of miR-1b, Let-7d, and miR-155. Transient transfection revealed that miR-155 significantly suppresses BDNF in H9c2 cells. Importantly, DSS colitis markedly decreased BDNF in both myocardium and serum. Levels of various proteins critical to cardiac homeostasis were also altered. Functional studies showed that BDNF increases cell viability and mitigates H2O2-induced oxidative damage in H9c2 cells, demonstrating its protective role in the adult heart. Mechanistically, cellular experiments identified IL-1 as the inflammatory mediator upregulating cardiac miR-155; this effect was confirmed in adult rats. Furthermore, IL-1 neutralizing antibody ameliorated the DSS-induced increase in miR-155 and concurrent decrease in BDNF in the adult heart, showing therapeutic potential. Our findings indicate that chronic colitis impairs heart function through an IL-1→miR-155→BDNF signaling axis. Introduction Inflammatory bowel disease (IBD) represents a spectrum of chronic inflammatory mediated conditions, including Crohn's disease and ulcerative colitis, manifesting in varying degrees and locations of colitis. While therapeutic research has been primarily targeted on affecting the pathways yielding abnormal colonic inflammation, IBD has known extraintestinal manifestations mediated through a chronic inflammatory and hypercoagulable state. Chronic inflammation has previously been identified as atherogenic with increased risk of developing cardiovascular disease. The interplay between patients with heart failure (HF) and consequential alterations in colonic integrity and mucosal bacteria burden has also been noted. In a Danish cohort study, patients with IBD had an increased risk of hospitalization for HF, a risk strongly correlated to periods of active disease. Patients with IBD diagnosed for greater than a year were noted to have an increased incidence of coronary artery disease events compared to controls despite having fewer risk factors for coronary artery disease. In addition, Crohn's disease and ulcerative colitis have been identified as independent risk factors for acute myocardial infarction. Despite the independent recognition of IBD as a risk factor for heart disease, the mechanisms mediating the changes in cardiac function in patients with IBD have not been well elucidated, partly due to unavailability of human heart tissue. Cardiac remodeling in the presence of chronic inflammation has been investigated as an important pathway eventually mediating physiologic HF. TNF-, an important inflammatory cytokine in the pathogenesis of IBD and a well-established therapeutic target for both Crohn's disease and ulcerative colitis, has been linked to left ventricle remodeling. While anti-TNF therapy is effective in reducing disease activity and inducing clinical remission in IBD patients, adverse events including heart failure have been observed. This suggests that TNF- signaling has a complex role in cardiac homeostasis and that other proinflammatory mediators such as interleukin 1beta (IL-1) or lipopolysaccharide (LPS), which are increased in IBD, could play a role in molecular remodeling in the adult heart. Clarifying how these inflammatory mediators induce cardiac remodeling may lead to development of novel therapeutic strategies. MiRNAs have been related to posttranscriptional regulation of gene expression in major cardiac physiological and pathological processes, and cardiac muscle phenotypes are tightly regulated by miRNAs to maintain cardiac homeostasis. miR-155 plays a key role in the homeostasis and function of the immune system, but abnormal high levels of circulating miR-155 found in patients with ulcerative colitis could be detrimental. While miR-155 knockout mice demonstrated resistance to developing cardiac hypertrophy, cardiac miRNA profiles in the presence or absence of chronic colitis have not been characterized. Whether and how miR-155 mediates cardiac remodeling in IBD remain to be determined. Brain-derived neurotrophic factor (BDNF) has been identified as a neurotrophin mediating wide-ranging effects on the cardiovascular system including cardiac and coronary artery development and maintenance of homeostasis including protection against reactive oxygen species. In adult mammals, BDNF governs autonomic transmission to the heart and exerts prominent angiogenic effects. Of note, the BDNF receptor, tropomyosin-related kinase receptor B (TrkB), exists in the myocardium. BDNF/TrkB signaling is required for the heart to fully contract and relax. Recent studies suggest that alterations in BDNF synthesis and function are associated with increased incidence of heart failure, arrhythmia, and myocardial infarction, indicating its necessity in the cardiovascular system. It is unclear, however, if chronic colitis suppresses BDNF by upregulating certain miRNAs. The aim of this study was to determine whether and how colitis alters miRNA profiles in the adult heart, and if changes in miRNAs are associated with alterations in functional proteins. Our results indicate that chronic colitis upregulates IL-1 augmenting certain cardiac miRNAs, particularly miR-155, which leads to suppression of BDNF corresponding to impaired heart function. Our findings reinforce the clinically noted connection of IBD and HF and suggest that cardiac impairment in IBD could be mitigated with IL-1 antibody. Cell culture H9c2 rat cardiaomyoblast cells were purchased from American type culture collection (ATCC, Manassas, VA) and maintained in Dulbecco's modified eagle medium supplemented with 10% fetal bovine serum and 0.1% penicillin-streptomycin solution. Transient transfection was performed using Lipofectamine RNAiMAX (Thermo Fisher Scientific, Waltham, MA). Animals Total 70 6-week old male Sprague Dawley rat littermates (150 to 200 grams) were purchased from Harlan (Indianapolis, IN) and used in the preclinical studies. The rats were housed in the UTMB animal facility with 12h light/12 dark cycle, a temperature range of 24-26C, and a relative humidity of 40-70%. Regular chow and water were provided ad libitum. Protocols for animal experiments. Three colitis models were used in this project. 1. Chronic colitis (n = 8 per group). Chronic colitis was induced with 3% DSS, which was applied for 4 cycles of 5 days each, with intermittent 9-day intervals of normal drinking water. 2. Acute TNBS colitis (n = 6 per group). Rats were lightly anesthetized with isoflurane (4% for induction and 1% for maintenance) and 250 l of TNBS in phosphate buffered saline containing 40% ethanol was injected intrarectally via a catheter, advanced to 8 cm into the colon. Control rats received one time intracolonical injection of 250 l of saline. 3. Acute DSS colitis (n = 6 per group). To determine if IL-1 antibody ameliorates DSS-induced miR-155 upregulation, 6-week old male Sprague Dawley rats were treated with 5% DSS in the drinking water for 7 days. IL-1 antibody (25 g/kg in 200 l of saline) was administrated daily by intraperitoneal injection for 7 days. Rats in control groups were subjected to daily intraperitoneal injection of 200 l saline. Animals were euthanized 3 hours after the last dose of IL-1 antibody. The individual hearts were collected, snap-frozen in liquid nitrogen, and pulverized for RNA extraction. All animals with experimental colitis were monitored daily by a member of the research team for assessment of food consumption and any unexpected discomfort or morbidity. At the end of each experiment, animals were decapitated under deep plane of anesthesia with 4% isoflurane. To assess the impact of IL-1 and TNF- on the miR-155 levels in the adult heart, 6-week old male Sprague Dawley rats were anesthetized with isoflurane (4% for induction and 1% for maintenance) and given recombinant IL-1 or TNF- (GeneScript, 10 g/kg) by tail vein injection. Control group received saline (n = 8 per group). Twenty-four hours later, animals were decapitated under deep plane of anesthesia with 4% isoflurane. Individual hearts were removed and snap-frozen in liquid nitrogen, followed by pulverization with mortar and pestle. A small aliquot of the pulverized cardiac tissue was used for isolation of total RNA containing miRNA. Ethics statement. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All procedures were approved by the Institutional Animal Care and Use Committee, The University of Texas Medical Branch at Galveston (Protocol # 1512071A). Rat echocardiography Rat cardiac function was examined via trans-thoracic echocardiography using a Vevo 770 High-Resolution Imaging System equipped with a 30 mHz probe (VisualSonics, Toronto, Canada). The rats were anesthetized with isoflurane (4% for induction and 1% for maintenance) and positioned in left lateral decubitus. The anterior chest hair was removed with Nair Hair Remover Lotion (Church & Dwight Co., Ewing, NJ). ECG was monitored throughout the experiment and temperature was kept at 37C on the heated plate. The left ventricular shortaxis and long-axis view and the apical four-chamber view were examined by two-dimensional and M-mode echocardiography. A number of cardiac function-related parameters were determined, including left ventricular ejection fraction (LVEF), LV mass, and LV size (short-axis area and long-axis length). Immunohistochemistry For histologic examination, a full-thickness colon specimen and half of the heart were obtained, fixed in 10% formalin, embedded in paraffin, sectioned, and de-waxed prior to histochemical staining stained with hematoxylin and eosin (H&E). For immunohistochemistry, five-micrometer sections were baked in a 50-55C oven for 1 hour. After antigen retrieval and one-hour blocking with 10% serum, the slides were treated with anti-COL3A1 mouse monoclonal antibody (sc-514601, Santa Cruz Biotechnology; 1:50 dilution) overnight, followed by washing three times 15 min each with 1X PBS. The sections were then incubated for 1 hour at room temperature with ALEXA-conjugated antibody (Invitrogen) diluted 1:400 in PBS. Images were captured using a LEICA DMI 6000 B microscope. Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay was performed using the TUNEL Assay Kit-HRP-DAB (ab206386, abcam, Cambridge, UK) and following the protocol provided by the manufacturer. Real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) Total RNA including miRNA was extracted using the miRNeasy Mini Kit (QIAGEN, Valencia, CA), followed by cDNA synthesis using SuperScript III First-Strand Synthesis System or Taq Myeloperoxidase (MPO) assay Frozen colon mucosa/submucosa was pulverized in liquid nitrogen, homogenized in 20 mM phosphate buffer (pH 7.4), and centrifuged at 4C for 10 minutes. Pellets were sonicated in 50 mM phosphate buffer (pH 6.0) containing 0.5% hexadecyl trimethyl ammonium bromide and centrifuged at 4C for 5 minutes. The supernatant (100 l) was incubated with 16 mM tetramethyl benzidine in 50% ethanol, 0.3 mM H 2 O 2, and 8 mM sodium phosphate buffer (pH 5.4) for 3 minutes. Myeloperoxidase activity was measured by reading the absorbance at 655 nm in a microplate reader. Subcellular fractionation Frozen tissue was smashed into fine powder in liquid nitrogen. Around 100 mg of pulverized tissue was transferred to a microcentrifuge tube and about 10 times tissue weight of ice-cold cytoplasmic extract buffer (250 mM Sucrose, 20 mM HEPES, 10 mM KCl, 1.5 mM MgCl 2, 1 mM EDTA, 1 mM EGTA, 1 mM DTT) containing protease inhibitors was added to each tube. The tissue was homogenized for 20 seconds with a Polytron homogenizer. After incubation on ice for 20 min, the nuclear pellet was centrifuged out at 720 g (3000 rpm) for 5 min. The nuclear pellet was washed once with the cytoplasmic extract buffer and resuspended with the nuclear extract buffer (20 mM Tris HCl, 420 mM NaCl, 1.5 mM MgCl 2, 0.2 mM EDTA, 10% glycerol, 0.1% SDS, 1 mM PMSF). The nuclei was incubated on ice for 40 min and vortexed on the highest setting for 15 seconds periodically, followed by centrifugation at maximum speed for 10 min. Measurement of B-type natriuretic peptide (BNP) and BDNF in serum and/or cytoplasmic/nuclear fractions Serum levels of BNP were determined using a B-type Natriuretic Peptide EIA kit (Cat. # RAB0386, Sigma, St. Louis, MO). BDNF levels in the serum and cytoplasmic/nuclear fractions were quantitated using a rat BDNF ELISA kit (Cat. # EK0308, ScienCell Research Laboratories, Carlsbad, CA). Cell proliferation assay Cell viability was assessed using the Vybrant1 MTT Cell Proliferation Assay Kit (Thermo Fisher Scientific). MicroRNA microarray MicroRNA microarrays were performed by LC Sciences (Houston, TX) using Microarray Version 21 (MRA-1003). Data were analyzed by subtracting the background and then normalizing the signals using a LOWESS (Locally-Weighted Regression) filter. The miRNA transcript was considered as reliably detectable only if the signal intensity was greater than 3 times the background standard deviation, the spot coefficient of variation was < 0.5, and at least 50% of the repeated probe signals were above the detection level. Statistical analysis All data were expressed as mean ± SEM. We used one-way analysis of variance (ANOVA) followed by Tukey post-hoc analysis for comparison of more than two means, and Student's ttest to compare between two means, and considered p<0.05 to be statistically significant. Chronic colitis impairs cardiac function To provide direct evidence that chronic colitis induces molecular remodeling in the adult heart, we first evaluated potential functional changes on the hearts of animals experiencing experimental colitis. Rats were exposed to 4 episodes of inflammation using 3% DSS to mimic the chronic, vacillating disease activity in IBD, a colitis model that has been very well characterized. Three percent DSS caused no mortality and had no significant effect on the body weight compared to controls (Fig 1A). To confirm the colonic activity of DSS, the colon length was measured, which was significantly shortened after the exposure compared to controls (17.1 ±0.57 vs 14.3±0.21, p<0.05) (Fig 1B). In the setting of colonic inflammation mediated by DSS, myeloperoxidase activity (5-fold, Fig 1C) and mRNA levels of IL-1 (9.5-fold), Mmp 7 (16-fold), and Mmp 9 (4.6-fold), but not Il18 (Fig 1D), were significantly increased (p<0.01). H&E staining revealed that DSS rats had significant alterations in the overall mucosal architecture (Fig 1E), including irregular crypts with variable diameters along the depth of single crypts or dilated crypts, bifurcation at the base of the crypt, and crypts herniated to the submucosa. Cryptitis, crypt abscesses, and erosions, i.e. the loss of surface epithelium, were observed in some areas. The DSS rats demonstrated a significant decline in LVEF (30%, Fig 1F) and accompanying increases in left ventricular mass (36%, Fig 1G) and size ( Fig 1H) compared to controls calculated on M-mode images from transthoracic echocardiography ( Fig 1I). These physiologic changes noted in chronic colitis were accompanied by an increase in serum levels of BNP (2.4-fold, Fig 1J), a biochemical marker for heart failure, which is strongly associated with LV hypertrophy. H&E staining of the heart sections showed cell swelling, irregular nuclear pattern, and enlarged spaces between muscle fibers in DSS rats (Fig 1K). MiRNA profiling identifies cardiac miRNAs significantly altered by DSS colitis In an effort to identify cardiac miRNA expression changes mediated by chronic colitis, miRNA microarrays were performed on the cardiac tissue of DSS and control groups (S1 Fig). A hierarchical cluster analysis was performed for all miRNAs with a signal intensity >32. A ttest was performed comparing control groups to DSS rats (S2 Fig). Sixty-eight miRNAs with high signal (>500) had significant changes in expression levels (p<0.05). Thirty miRNAs were significantly increased (Table 1) and 38 were decreased (S1 Table) (p<0.05). Fifty-two miR-NAs with low signal (<500) also had significant changes, among which 26 miRNAs including miR-155 were upregulated and 26 miRNAs downregulated (p<0.05). The data was deposited in the GEO Repository (Accession No. GSE158991. URL: https://www.ncbi.nlm.nih.gov/geo/ query/acc.cgi?acc=GSE158991). MiRNAs putatively targeting the BDNF gene were predicted and confirmed by RT-qPCR Using microRNA.org, miRDB, and miRWalk, target genes of all significantly altered miRNAs by DSS colitis were predicted. Interestingly, a total of 8 miRNAs increased by colitis in microRNA Loss of BDNF in rat models of experimental colitis In order to explore if changes in cardiac miRNA are associated with BDNF reduction, we examined the alterations in expression of BDNF, as well as notable cardiac proteins in DSS rats compared to age-matched controls: -catenin, whose re-activation in cardiomyocytes triggers hypertrophic response via activation of mitogen-activated protein kinases ; BCL2, which has both pro and anti-apoptotic activity in heart failure ; phosphorylated AKT (pAKT), which is associated with short term increases in contractile function and efficient oxygen utilization in the myocardium ; STAT3, which is a downstream regulator of gp130 signaling associated with compensatory hypertrophy during periods of cardiac pressure overload ; COL3A1, which is a marker of cardiac fibrosis; ELAV-like protein 1 (ELAVL1), which is an important mediator of pyroptosis in inflammatory mediated heart failure ; and finally cleaved Caspases 3 and 7, whose enzymatic activity is necessary to complete apoptotic cell death. In chronic colitis, BDNF protein levels in the myocardium were significantly reduced, accompanied by decreases in GSK-3, BCL2, pAKT(S133) and increases in -catenin, STAT3, COL3A1, and ELAVL1 compared to that in age-matched healthy controls (Fig 4A and 4B). Accumulation of COL3A1 in the DSS rat heart was confirmed by immunofluorescence, suggestive of cardiac fibrosis (Fig 4C). TUNEL staining showed very few positive apoptotic cells in both control and DSS rats (Fig 4D). Subcellular fractionation followed by enzyme-linked immunosorbent assay revealed that both cytoplasmic and nuclear BDNF levels in the adult heart were significantly attenuated by DSS (43% and 58%, respectively, p<0.05) (Fig 4E and 4F). This was accompanied by a significant reduction in circulating BDNF (91%, p<0.01) (Fig 4G). DSS induced colitis also suppressed BDNF mRNA expression in the adult heart (30%, p<0.05) (Fig 4H). In order to validate the changes induced by DSS, we also assessed BDNF expression in TNBS-induced acute colitis in adult rats, another well characterized rodent model of colitis. BDNF protein levels in the serum and mRNA expression in the heart were significantly reduced by TNBS-induced colitis compared to controls (95% and 33%, respectively, p<0.05) (Fig 4I and 4J), confirming that both acute and chronic inflammation in the colon suppresses BDNF levels in the heart. BDNF ameliorates H 2 O 2 -induced apoptosis in H9c2 rat cardiomyoblast cells To help establish further the functional importance of BDNF in the adult heart, we investigated if BDNF impairs apoptosis in cardiomyocytes. While recombinant BDNF protein had no effect on BDNF mRNA levels, the rat BDNF-specific siRNA was successfully transfected into H9c2 rat cardiomyoblast cells with a significant reduction in BDNF mRNA (70%, p<0.01) (Fig 5A). Cell proliferation quantified by MTT colorimetric assay was significantly increased in the cells treated with recombinant BDNF protein compared to control cells (31%, p<0.05). A significant reduction in cell proliferation was noted in the BDNF knockdown group (35%, p<0.01 vs. siControl group) (Fig 5B). Apoptosis in H9c2 cells was induced through the administration of H 2 O 2, manifested through cleavage of Caspase-3 and elevation of phosphorylated histone H2AX (H2AX) (Fig 5C, left plate); the changes were mitigated by BDNF overexpression (Fig 5C, right plate). These findings demonstrate a protective role of BDNF in cardiomyoblasts. IL-1 increases miR-155 and decreases BDNF levels in the adult rat heart To determine how systemic inflammation influences miR-155 expression in cardiac myocytes, independent experiments were performed to identify mediators of chronic inflammation augmenting myocyte expression of miR-155. First, H9c2 cells were treated with inflammatory mediators H 2 O 2, TNF-, IL-1, or low and high dose LPS. Only IL-1 upregulated miR-155 levels significantly (1.7-fold, p<0.01) (Fig 6A). Elevated levels of miR-155 responded in a dose dependent manner up to concentrations of 20 ng/mL (10 and 20 ng/mL p<0.01 vs. concentration zero) (Fig 6B). To verify that IL-1 stimulates miR-155 in rat myocardium, we treated adult rats with recombinant rat IL-1 or TNF- and evaluated miR-155 levels in cardiac tissue by RT-qPCR. These in vivo studies demonstrated that IL-1, but not TNF-, augments miR-155 in the adult rat hearts compared to vehicle treated controls (2.4-fold, p<0.01 vs. control and TNF-) (Fig 6C). To further confirm that IL-1 alters miR-155 expression in the chronic colitis model, IL-1 neutralizing antibody was concurrently administered in DSS rats. IL-1 neutralizing antibody negated the DSS-induced upregulation of miR-155 (Fig 6D) in the adult rat heart (p<0.01 vs. vehicle-treated DSS rats). More importantly, downregulation of BDNF by DSS was concurrently abrogated by administration of IL-1 neutralizing antibody (Fig 6E). Discussion Population-based studies found that IBD is associated with increased risk of hospitalization for heart failure, developing ischemic stroke, and myocardial infarction. More recent studies have identified extracardiac sources of pro-inflammatory cytokines which can induce cardiac pump dysfunction. Circulating inflammatory mediators, such as TNF-, have been noted to correlate with disease severity in HF patients, and elevated levels are prognosticators of poor clinical outcomes. In addition, patients presenting with HF with the highest quartile of circulating IL-1 had increased all-cause mortality at one year compared to their comorbidity matched cohorts. The etiology of this chronic inflammatory state has not entirely been elucidated, although is thought to be mediated in part by increased release of endotoxins originating from the intestines. In this study, we showed functional and anatomical changes of reduced LVEF corresponding with hypertrophic ventricular changes using trans-thoracic echocardiography in rats with chronic colitis, after demonstrating baseline echocardiographic results consistent with previously described rat controls. These findings corresponded with biochemical evidence of heart failure with a 2.5 fold increase in BNP from control rats. This supports the population-based data that colitis can induce more severe phenotypes of HF and reinforces a connection between colonic integrity and extracolonic consequences in the heart. More importantly, we identified IL-1 as a major regulator of cardiac miR-155, which in turn suppressed heart BDNF. The significance of IL-1 in the chronic colitis model was reinforced when DSS-induced augmentation of miR-155 expression was shown to be reversed with IL-1 neutralizing antibody. These findings indicate that IL-1 is one of the missing links between IBD and HF. Blocking IL-1 might benefit patients with IBD by preventing cardiac remodeling that could lead to heart failure. Theoretically, IL-1 neutralization could also benefit patients with chronic autoinflammatory diseases such as rheumatoid arthritis, as well as type 2 diabetes, which is also considered as a chronic lowgrade inflammatory disease. We previously identified miR-155 as an important modulator of epithelial integrity in the setting of chronic colitis via its interference of colonic tight junctions by decreasing E-cadherin expression. Let-7 has significantly altered expression in both Crohn's disease and ulcerative colitis, albeit in contradictory patterns. In cardiovascular disease, in addition to the before mentioned identification of miR-155 as a potential biomarker for non-ischemic cardiomyopathy, Let-7 has well established alterations of expression patterns in various cardiovascular diseases, notably upregulated in heart failure and ventricular hypertrophy. miRNA profiling of the mouse adult hearts revealed a significant proportion of miR-1 of known miRNA reads. When exposed to a cardiomyocyte-specific deletion of a gene required for miRNA production, LV malformations leading to dilated cardiomyopathy with subsequent LV dysfunction (manifested by diminished fractional shortening) were noted. We utilized microarray analysis to identify the rat miRNAs significantly modulated by DSS colitis, and isolated the mRNAs predicted to target BDNF. Three miRNAs, miRNA-155, Let-7, and miR-1, were significantly increased in DSS colitis, and their common target, BDNF, was decreased significantly in cells transfected with the miRNA of interest. Prominent was miR-155, which was upregulated by IL-1 both in vitro and in vivo. BDNF represents an emerging target for cardiovascular research due to its diverse effects of heart centric homeostasis with alterations in expression in this growth factor implicated in a wide variety of cardiovascular diseases. We demonstrated that in response to chronic colitis, both serum and heart BDNF levels were significantly downregulated, which could adversely influence heart function. Importantly these changes were accompanied by alterations in protein expression of mediators which collectively are associated with molecular and biochemical modifications leading to hypertrophy (Fig 1K), fibrosis (Fig 4C), and the promotion of HF. BDNF knockdowns demonstrated significant reductions in cell proliferation in cardiomyoblast cells (Fig 5). Conversely, forced expression of BDNF reduced H 2 O 2 -induced apoptosis. In total, these findings support BDNF's role as a mediator of chronic colitis-induced cardiac dysfunction. The DSS model of induced colitis is considered a good preclinical model that exhibits many phenotypic features of relevance to human IBD. DSS mice have increased anxiety-like behavior and decreased electroretinography amplitudes, a measurement of retinal function, suggestive of two common extraintestinal manifestations. Our findings indicate that the DSS rat model could be a useful tool for mechanistic elucidation of cardiovascular diseases in IBD. However, rodent DSS models have their limitations as no animal models can fully recapitulate human IBD phenotypes and symptoms. Another limitation in this study is that the exact origins of miR-155 measured in the hearts remain to be determined. While our results suggest that IL-1 elevates miR-155 in cardiomyocytes, this event might also occur in the colon. High levels of miR-155 of gut origin could be transported to the heart by exosomes since miR-155 was found to be the most highly expressed miRNA in the blood samples of ulcerative colitis patients. miR-155 is also highly elevated in the colon mucosa and feces collected from Crohn's disease and ulcerative colitis patients. While this study helps shed light on cardiac remodeling in IBD, we are aware that the human heart differs from the murine heart. Future studies could include procuring human heart tissue from deceased IBD patients and deceased control subjects without IBD, and comprehensive characterization of their molecular differences. Our findings suggest that IL-1 plays a major role in mediating cardiac remodeling through IL-1!miR-155!BDNF signaling axis. IL-1 neutralization might be further tested for its ability to ameliorate HF in different preclinical models of colitis before consideration of proceeding to human clinical trials. While BDNF supplementation and miR-155 antagonists should be explored for their therapeutic potential in mitigating IBD-induced cardiac remodeling/HF, we cannot rule out other factors that may contribute to cardiac remodeling in IBD. For example, studies demonstrated a significant reduction in the diversity of the stool microbiome of individuals with IBD. The microbiota of patients with IBD is characterized by depletions in butyrate-producing bacteria with anti-inflammatory effects and an expansion in pathogenic bacteria. While inflammation can be well controlled by drug therapy or surgery, gut microbial dysbiosis persists and may have a life-long impact on IBD patients. Thus, future studies should also investigate how colitis-induced microbiota dysbiosis influences the cardiac system. In summary, the present study demonstrated that chronic colitis induced by DSS suppresses BDNF, a cardiac modulating neurotrophin with wide-ranging effects on normal cardiovascular development. This change was accompanied by the altered expression of proteins associated with biochemical and structural alterations associated with HF. We confirmed that BDNF ameliorated H 2 O 2 -induced apoptosis in H9c2 cardiomyoblast cells. We noted that three miRNAs, particularly miR-155, were predicted to be and then confirmed as regulators diminishing BDNF expression. Finally, we mechanistically connect chronic colitis-induced inflammation and miR-155-mediated changes in cardiomyocytes through IL-1. These findings, in summation, provide an explanation to the previously noted relationship between HF and IBD. This supports the idea that chronic inflammation, and the modulation of its molecular byproducts, continues to represent a compelling area of therapeutic investigation particularly in patients with comorbid cardiovascular disease and colitis.

Texture Plus Depth Video Coding Using Camera Global Motion Information In video coding, traditional motion estimation methods work well for videos with camera translational motion, but their efficiency drops for other motions, such as rotational and dolly motions. In this paper, a motion-information-based three-dimensional (3D) video coding method is proposed for texture plus depth 3D video. The synchronized global motion information of the camera is obtained to assist the encoder improve its rate-distortion performance by projecting the temporal neighboring texture and depth frames into the position of the current frame, using the depth and camera motion information. Then, the projected frames are added into the reference buffer list as virtual reference frames. As these virtual reference frames could be more similar to the current to-be-encoded frame than the conventional reference frames, the required bits to represent the residual will be reduced. The experimental results demonstrate that the proposed scheme enhances the coding performance for all camera motion types and for various scene settings and resolutions using H.264 and HEVC standards, respectively. With the computer graphic sequences, for H.264, the average gain of texture and depth coding are up to 2 dB and 1 dB, respectively. For HEVC and HD resolution sequences, the gain of texture coding reaches 0.4 dB. For realistic sequences, up to 0.5 dB gain (H.264) is achieved for the texture video, while up to 0.7 dB gain is achieved for the depth sequences.

Q: If I invest in a company that goes bankrupt, is that a gain or a loss? This is a real situation, but I will use fake numbers. Let's say I invest $10,000 in an investment trust and my shares eventually appreciate to the point where they are worth $60,000. This is not an equity position, but a debt position because my shares represent assets that are supposed to be held by the trust. Then something bad happens and the company goes bankrupt. The bankruptcy trustees assess the value of my shares at $60,000 (the amount the company owes me) and then announce that they are liquidating the company and paying out 1/3 of par value to the creditors, therefore I will receive $20,000. In this case I will ultimately get returned to me more than I originally "loaned" to the company, but I will lose a lot of what they owed me. Legally speaking I am a "creditor" of the company, not a "shareholder". In this case, from a tax perspective (United States), have I suffered a gain or a loss? (As an analogy, suppose a bank loans $500,000 to a business and according to the terms of the loan, the business ends up owing $600,000 including interest. But then suppose the business goes bankrupt and after the liquidation the bank is only paid $540,000. Has the bank had a $40,000 gain or a $60,000 loss? This is analogous to my situation.) A: I'll give the credit to @Quid in the comments section of the question. You put out $10k, you got back $20k, that's a cash gain of $10k, how the asset was valued between your purchase and sale isn't relevant. From an accounting perspective, the company is the only party that is realizing the loss (as they have sold the asset for 40K less than par). You the buyer, only get to see the initial buy and sale of such capital asset. Example: A company purchases a car for $20,000 and after depreciation it is worth (book valued at) $2,000. It is then sold to a customer for $3,000. Does the customer realize a loss of $1,000? No. Does the company realize a gain of $1,000? Yes. Your bank analogy is flawed in two ways: We don't know how long this has transpired. If this all occurred in the course of a day, the bank would claim a 40K gain. We don't know if interest was included in contract. Some contracts include "You will pay 100K, in interest, when this comes due." If this holds true and the same example occurred, the bank would record a 60K loss.

Fragmented Landscape of European Policies in the Energy Sector: First-Mover Advantages In order to achieve the commonly agreed emission reduction target, the European Commission developed binding national targets for each member state until 2030 and called upon the member states to submit National Energy and Climate Plans to ensure increased transparency for the respective national targets and strategies. An analysis of these plans shows that some of the emission reductions set at the national level prescribe a more ambitious decarbonisation than the EU-wide limits. However, since a transformation to a climate-friendly system requires considerable investment, the question arises as to why some states apparently want to be in the vanguard. We find that countries may have an incentive to outperform other states in the development of a low-carbon electricity system in order to pass on part of the transformation costs to neighbouring countries.

NEW YORK — Tory Burch was inspired by higher learning for her fall collection at New York Fashion Week. Specifically, she looked to Black Mountain College, a progressive school in North Carolina. Budding star Lana Condor of Netflix’s “To All the Boys I’ve Loved Before” also attended during her very first fashion week.

First record of a bloom of the invasive species Ceratium furcoides (Levander) Langhans 1925 in Rio Grande do Sul state, Brazil. Schrank is dinoflagellate genus commonly found in marine environments, and considered invasive in continental waters (Silva et three different depths al., 2012). Species of this genus are able to develop blooms due to their mobility and resistance to sedimentation, to the occupation of the surface layer and optimised use of resources such as light and nutrients ( ). Even though its blooms are not toxic, they still bring harmful effects to aquatic communities as anoxic conditions causing the death of a local population of lobsters (Pitcher and Probyn, 2011).In Brazil, the occurrence of a species of this genus has been recently registered in freshwater ecosystems. Santos-Wisniewski et al. recorded a bloom of

Projected changes in wind speed and wind energy potential over West Africa in CMIP6 models The development of wind energy in West Africa is essential to meet the rising energy needs due to population growth and societal development. However, only few studies have investigated the changes in turbine hub-height wind characteristics over the region under changing climate. This study aims at assessing the impact of climate change on wind power density (WPD) over West Africa using the simulations from the newly developed Coupled Model Intercomparison Project version 6 (CMIP6) models. The CMIP6 near-surface wind speed and directions simulations for the historical climate (19852014) were compared with ERA5 reanalysis data using multiple descriptive statistics. Relative to ERA5 reanalysis, the CMIP6 models alongside their multimodel ensemble mean (EnsMean) realistically reproduce the near-surface wind characteristics (i.e. wind speed and directions) across most subregions of West Africa, although noticeable biases still exist. Overall, the CMIP6 EnsMean performs better than most individual models at capturing the near-surface wind speed over the region. Under global warming, we find a robust projected increase (about 70%) in WPD over the Guinea coast subregion of West Africa, especially in JuneJulyAugust season. The DecemberJanuaryFebruary and MarchAprilMay seasons show alternating projected WPD increase and decrease, with predominantly robust projected decrease over the Sahel subregion. The projected increase over the Guinea coast has strong temporal qualities, with the end of the century (20702099) changes showing stronger magnitude compared to the mid-century (20402069) changes, and thus may provide a commercially viable renewable energy source.

Human Cues in Self-help Lifestyle Interventions: an Experimental Field Study Background: Self-help eHealth interventions are generally less effective than human-supported ones, as they suffer from a low level of adherence. Nevertheless, self-help interventions are useful in the prevention of non-communicable diseases, as they are easier and cheaper to widely implement. Adding humanness in the form of a text-based conversational agent (TCA) could provide a solution to non-adherence. In this study we investigate whether adding human cues to a TCA facilitates relationshipbuilding with the agent, and makes interventions more attractive for people to adhere to. We will investigate the effects of two types of human cues, which are visual cues (eg, human avatar) and relational cues (eg, showing empathy). Objective: We aim to investigate if adding human cues to a TCA can help increase adherence to a self-help eHealth lifestyle intervention and explore the role of working alliance as a possible mediator of this relationship. Methods: Participants (N=121) followed a 3-week app-based physical activity intervention delivered by a TCA. Two types of human cues used by the TCA were manipulated, resulting in four experimental groups, which were visual cues-group, relational cues-group, both visual and relational cues-group, and no cues-group. Participants filled out the Working Alliance Inventory Short Revised form after the final day of the intervention. Adherence was measured as number of days participants responded to the messages of the TCA. Results: One-way ANOVA revealed a significant difference for adherence between conditions. Against our expectations, the groups with visual cues showed lower adherence compared to those with relational only or no cues (t = -3.415, P =.001). No significant difference was found between the relationaland no cues-groups. Working alliance was not affected by cue-type, but showed to have a significant positive relationship with adherence (t = 4.136, P <.001). Conclusions: We hypothesize that the negative effect of visual cues is due to a lack of transparency about the true nature of the coach. Visual resemblance of a human coach could have led to high expectations that could not be met by our digital coach. Furthermore, the inability of TCAs to use non-verbal communication could provide an explanation for the lack of effect of relational cues or the effect of cue-type on working alliance. We give suggestions for future studies to test these potential mechanisms. Clinical Trial: Pre-registration: OSF Registries, https://osf.io/mgw2s https://preprints.jmir.org/preprint/30057 JMIR Preprints Cohen Rodrigues et al (JMIR Preprints 29/04/2021:30057) DOI: https://doi.org/10.2196/preprints.30057 Table of Contents Human Cues in Self-help Lifestyle Interventions: an Experimental Field Study Original Manuscript agent; chatbot; adherence; working alliance. Non-communicable diseases (eg, cardiovascular diseases, type-2 diabetes) are the leading cause of death globally. Engaging in a healthy lifestyle can help in the treatment and prevention of many of those diseases. This could be facilitated by eHealth, which are digital tools that can be used by a healthcare professional to provide remote support, or that can provide automated support. Studies show that eHealth is effective in improving lifestyle behaviors, and in the prevention and treatment of non-communicable diseases. Automated self-help interventions are easier and cheaper to widely implement as they require no interference of healthcare professionals, who indicate to experience barriers in lifestyle support such as a lack of time or insufficient experience with lifestyle support. However, meta-analyses show that human-supported digital lifestyle interventions are more effective than self-help ones. Adherence, or the extent to which a person uses the eHealth intervention as intended, is a problematic within self-help interventions. As intervention adherence is related to more positive health outcomes, finding ways to make people adhere to self-help interventions would be necessary to reach optimal effectiveness. Although human-supported interventions generally outperform self-help ones, this does not necessarily imply the support of a healthcare professional. Meta-analyses revealed that contact with a nonprofessional or administrative support by a human being is enough to both ensure intervention effectiveness and prevent people from dropping out of the intervention. It seems that some level of "humanness" rather than professional guidance is the key ingredient within human-supported interventions. The underlying reason why people would like a level of humanness into the intervention, could be the need of a personal relationship with their care provider. This so-called working alliance can be defined as the level of agreement on goals that are set for treatment, on tasks that must be performed to reach this goal, and the relational bond between healthcare professional and patient. Working alliance with a human care provider is a predictor of intervention adherence and effectiveness both in regular face-to-face and in digital therapy or treatment. However, people are also able to form relationships with computers. People interact with computers as they would do with human beings, and apply similar social rules and heuristics. Studies show that people can also develop a working alliance within fully automated digital interventions, and that this leads to more positive treatment outcomes. In self-help interventions, humanness can be added by the use of a so-called conversational agent (CA). These computer-based agents can mimic human-like conversational behavior (eg, respond to input, generate output, apply turn-taking), and be used to provide automated support. An embodied conversational agent (ECA) is visually present on screen and can provide non-verbal cues (eg, hand gestures), while a text-based conversational agent (TCA) is able to communicate with text only. A TCA has the advantage of being easier to develop, being easier to apply in a mobile app, and therefore being more suitable for widespread implementation. Studies show that people show more relational behaviors or feel more social presence when they believe that their interaction partner is a human being rather than a computer. To enhance these perceptions while interacting with CAs, human cues could be applied, such as an avatar of a human being, a human tone-of-voice, or lower speed of feedback. Furthermore, non-verbal communication can be replaced by adding emoticons. Besides the appearance of the messages and CA, human cues could be applied to the content of its messages and behavior. It is possible to add human conversation rules in computer-generated conversations, such as humor, empathy, and small talk, which are often used by humans to establish a relationship. Studies with ECAs show that applying such human cues to the interaction increases the working alliance users experience with the ECA and their intention to use the ECA. Although studies about digital interventions with TCAs have been conducted before, only a small amount has focused on their application in lifestyle change interventions. Additionally, the effects of human cues are predominantly tested with ECAs. Therefore there is little knowledge about how human cues affect people's relationship with TCAs, or their adherence to TCA-supported interventions. Furthermore, the majority of studies test the effects of either using human cues or not, while we are interested in the effects of two different types of human cues and how these interact. In this study we will focus on both the appearance and behavior, or in other words, the visual and relational cues, that can be integrated into a TCA to increase the level of humanness. We predict that these will improve the working alliance people experience with the TCA, and in turn their adherence to the intervention. To test this, we conducted a field-experiment in which people followed a 3-week app-based physical activity intervention with automated support from a TCA. We manipulated the cues used by the TCA, and measured the working alliance that people experience with the TCA, and the number of days they adhere to the intervention. This allows us to answer the following research questions: RQ1: Is there an effect of human cue type (visual and relational cues) on adherence? RQ2: Does working alliance mediate the effect of human cue type (visual and relational cues) on adherence? Study Design The three-week field experiment was conducted in March and April 2020. We employed a 2 (visual cues: yes, no) x 2 (relational cues: yes, no) between-subjects design. There were four experimental groups, in which the TCA used visual and relational cues, visual cues, relational cues, or no human cues. Participants We recruited (N = 269) healthy participants between 18 and 30 years old with flyers on the university campus and via social media, who were willing to work on their level of physical activity, had access to a smartphone running iOS or Android, and were sufficiently proficient in English. We excluded participants who were not able to engage in a normal physical activity pattern using the Physical Activity Readiness Questionnaire (PAR-Q). Students from Leiden University received credits (required to complete their first year) for their participation, and all participants who would complete the study would enroll in a lottery (with the chance of winning one of the three Fitbit devices, or one of the 100 vouchers worth €10,-). Power calculations (G*Power) identified a minimum sample size of 128 to detect a medium between-group effect (f =.25) of cue-type with an alpha of.05 (ANOVA with 4 groups). Given the high attrition rates in similar studies (eg, ), we aimed to recruit about double the required number of participants. All participants provided their consent before the start of the experiment, and the study was approved by the Psychology Research Ethics Committee of Leiden University. Intervention The aim of the intervention was to enhance participants' physical activity levels by increasing daily step counts. The intervention consisted of daily exercises, such as a quiz about the health consequences of physical activity and a decisional balance worksheet (see Appendix 1 for overview of daily exercises). These exercises would take about 5 to 10 minutes each day to complete. The exercises were based on behavior change techniques (BCTs), such as prompts, information about health consequences, review of goals, and social reward. These are intervention components designed to regulate behavior by reinforcing factors that facilitate behavior change, and mitigating factors that hinder behavior change. These behavior change techniques were incorporated following the Transtheoretical Model of health behavior change, which views behavioral change as an upward spiral process involving progress through five stages (i.e. pre-contemplation, contemplation, preparation, action, maintenance). The model has been used to target a wide range of health behaviors. Technical Implementation of the Benefit StepCoach for Android and Apple's iOS Platforms The Benefit StepCoach app was implemented with MobileCoach (www.mobile-coach.eu), an open-source software platform for smartphone-based and chatbot-delivered behavioral interventions (eg, ) and ecological momentary assessments (eg, Appropriate interactions were implemented, i.e. asking participants for their permission, to allow the apps to access the step data. Moreover, each experimental group was assigned a dedicated TCA. Text-based Conversational Agent Participants interacted daily with a TCA, the virtual coach who delivered the intervention and offered various conversational turns. Via the chat feature, the TCA delivered daily exercises in line with the intervention and would respond to messages of the participants via conversational turns (see Figure 1). All conversational turns were scripted. Each day would consist of two to four conversational turns. The first message would be sent in the morning (9:00 am), and the following messages after a reply of the participant. If the participant would not have replied yet, the TCA would send a reminder in the afternoon (3:00 pm). Across the experimental groups, the intervention (eg, tasks and feedback) was identical, but the conversational turns differed in the type of cues the TCA used. We manipulated two types of human cues: visual cues, which were related to the humanness of the communication style, and the design and appearance of the messages (human avatar, use of emoticons, human tone-of-voice, and response delay), and relational cues, which were related to the content of the messages, and to what extent these followed social scripts and human conversation rules (eg, showing empathy, self-disclosure, humor, small talk, and meta-relational communication) (see Figure 2). Adherence Adherence was measured based on the number of days participants finished the session of conversational turns. Participants were marked as "adherent" for a particular day if they had replied to the final message of the TCA before the end of the day (12:00 pm at midnight). Given the duration of the intervention, the level of adherence over the whole study could range between 1 and 21 days. Physical Activity We measured physical activity through objective step count data retrieved from Apple Health or Google Fit (depending on the smartphone of the participant). Effectiveness was based on the baseline average step count in the week before the intervention, as well as on the step counts retrieved during the intervention itself. To assess baseline levels of physical activity, the International Physical Activity Questionnaire Short Form (IPAQ-SF) was used. The questionnaire consists of seven items asking the participants about their time spent on vigorous and moderate physical activities, walking, and sitting during the previous week. The output is a MET score, representing the amount of energy used to carry out the reported physical activities. The IPAQ-SF has been shown to have a high reliability, but minimal validity. Therefore we decided to additionally use objective step count as baseline measurement. Working Alliance Working alliance with the TCA was measured with an adjusted version of the Working Alliance Inventory Short Revised form (WAI-SR). The WAI-SR consists of 12 items measured on a 5-point Likert-type scale ranging from 1 (seldom) to 5 (always), subdivided in 3 subscales: agreement on tasks, agreement on goals, and bond. Questions were adjusted to fit the context of the study by using the words "coach", "lifestyle" and "intervention" (eg, "The coach and I collaborate on setting lifestyle goals."). The WAI-SR has been shown to have sufficient reliability and validity, and our adjusted version showed to have a high internal consistency (Cronbach's =.945). Procedure A week before the start of the intervention 282 participants provided digital informed consent and filled in a screening survey assessing the inclusion and exclusion criteria. 226 eligible participants received a link to the iOS or Android app store to download the Benefit StepCoach app. Once the app was downloaded, participants were asked to go through the onboarding procedure to correctly configure the app (eg, allowing push messages and access to step count data via Apple Health or Google Fit), and to complete the baseline survey. Participants were reminded through emails and text messages to complete the onboarding and baseline survey (measuring demographics and baseline characteristics) after 3, 4 and 5 days, and excluded if they did not do so before the start of the intervention. Participants were allocated to one of the four conditions by an automated mechanism within the app. All participants started simultaneously with the three-week (21 days) intervention. Each day, the TCA would send the participants one or several short exercises to complete that day (eg, quiz or worksheet, see Appendix 1 for overview of daily exercises) via a push notification. After completing the final survey on day 22 (measuring Working Alliance), participants would receive the debriefing. Data Analysis The analyses were preregistered via the Center for Open Science. Intervention effectiveness independent of cue-type was tested using a one-tailed dependent samples t test in which we compared average step count during the baseline week with the final week of the intervention. For the first research question (Is there an effect of human cue type (visual and relational cues) on adherence?), we predicted that the condition with both visual and relational cues would lead to the highest adherence, followed by the conditions with either visual or relational cues, and the condition with no human cues. To test this, a one-way between-subjects ANOVA with planned contrasts was conducted. Because the differences in mean adherence of each group were different than expected, the planned contrasts we ran were different from those pre-registered. In the first alternative post-hoc analysis we compared the visual & relational cues-and visual cues-conditions with the relational cues-and no cues-conditions. In the second, we compared the relational cues-condition with the no human cues-condition. For the second research question (Does working alliance mediate the effect of human cue type on adherence?), we predicted that working alliance with the TCA would be highest in the condition with both visual and relational cues, followed by the conditions with either visual or relational cues, and then the condition with no human cues. In turn, we expected that a higher working alliance would lead to a higher adherence. A one-way between-subjects ANOVA was conducted to compare the working alliance with the TCA between the four different conditions. As we did not find significant differences, we did not conduct our preregistered mediation analysis. We did run an additional regression analysis with working alliance as independent and adherence as dependent variable. Statistical analyses were conducted with SPSS (version 26; IBM Corp) using a p-value of.05 as the level of significance. Table 1). Intervention Effectiveness In our preregistration, we proposed to test hypotheses with effectiveness as outcome variable. Our power calculations identified a minimum sample size of 128 to detect the expected effects of experimental group on effectiveness. However, as we had insufficient cases with both a valid baseline step count and a minimum of 5 days of step count registered in the final week, we did not have enough power to detect this effect. We therefore decided to focus in this paper on adherence as outcome variable, and report the analyses with effectiveness as outcome variable in Appendix 2. To test whether the intervention was effective (independently of the experimental condition), Adherence We found a significant difference in adherence between the conditions, F = 3.901, P =.011 (see Table 2 for mean and SD per group). By visually inspecting the means, we saw that the differences between groups were not as expected (see Figure 3). The contrast analyses showed that in the relational cues-and no cues-conditions there was a significantly higher adherence than in the other two conditions, t = -3.415, P =.001. However, adherence in the relational cues condition was not higher than in the no human cues condition, t=.458, P =.65. So contrary to what was expected, participants were less adherent to the intervention in the groups in which the TCA used visual cues compared to the groups without visual cues. Furthermore, when the TCA used relational cues, participants were not more adherent than when the TCA used no human cues at all. per experimental condition, with 95% confidence intervals, including the post-hoc contrast between groups with and groups without visual cues. Working Alliance There was no significant difference in working alliance between the conditions, F = 1.110, P =.35 (see Table 2 for mean and SD per group). However, we did find a positive relationship between working alliance and adherence, b =.202, t = 4.136, P <.001. These outcomes indicate that adding human cues did not lead to a better (or worse) reported working alliance with the TCA, but that at the same time, participants who reported a better working alliance were more adherent to the intervention. DISCUSSION We investigated if and how a TCA could help increase adherence to a self-help eHealth lifestyle intervention. Regarding our first research question (Is there an effect of human cue type (visual and relational cues) on adherence?), the results of our field experiment showed that, contrary to our expectations, the use of human cues by the TCA did not lead to a higher adherence. In contrast, visual cues even led to a lower adherence. To answer the second research question (Does working alliance mediate the effect of human cue type (visual and relational cues) on adherence?), we found that human cues did not lead to a higher working alliance with the TCA. We did, however, find that a better reported working alliance was related to a better adherence to the intervention. Our results did not show the positive effects of human cues and visual elements that have been reported in previous studies. On the contrary, we even found a negative effect on intervention adherence for visual cues. One reason for this could be that, before the start of the intervention, we did not tell participants whether they would be coached by a human being or a computer. This lack of transparency might have led to expectations that could not be met by the TCA, which might have led to frustration among users. Many studies however show a positive effect on user perceptions and user behavior of not disclosing the nature of an automated chatbot, or of suggesting that users are interacting with a human being while they are not. However, Mozafari and colleagues show that the effects of disclosure depend on whether there are errors in the conversation with a chatbot. In their study with a customer-service bot, they found that when the chatbot was not able to solve a customer's issue, the negative responses to these errors could be prevented by disclosing the chatbots true nature beforehand. Although our study concerned a lifestyle intervention, similar effects could have occurred. As visual cues might have wrongly suggested communication with a human being and our CA was not always able to respond fully correctly (as the messages were preprogrammed), correctly informing participants about the nature of the agent could have prevented the negative effects of the errors within the conversations. Furthermore, the avatar we used in the visual cues conditions might have played a role. We intentionally chose a younger and healthy-looking female agent to resemble the psychology student population, and because a young female peer agent is generally preferred in health coaching tasks. However, some literature suggest that male agents are preferred as athletic trainer, which might have influenced the results if our participants perceived the TCA to be an athletic coach rather than a health coach. Furthermore, another study shows that non-ideal overweight agents are seen as more trustworthy and lead to higher use intentions, which suggests our TCA might have been too slender and healthy looking. Additionally, similarity between CA and target population can have a downside when the agent is perceived as unhelpful. The user's perception of the helpfulness of the agent depends on the goal of the user, and as our participants might have participated in the intervention with other goals than increasing physical activity (eg, gaining participant-credits), the similar-looking agent might have been unhelpful to them, leading to a lower adherence. All in all, the negative effect of the visual cues on adherence could therefore have been due to a lack of transparency about the true nature of the CA, and the type of visual cues we applied to our CA. The absence of an effect for relational cues in our study contradicts previously mentioned studies in which a positive effect was found. However, what is important to note is that these studies concern ECAs, while we used a TCA. ECAs generally outperform text-based ones, which can be explained by an additional range of design characteristics an ECA can make use of. In one study though, there was no difference found between a TCA and an ECA, which the authors argued was due to the lack of incorporating non-verbal communication in the latter one. Finally, we found that people who reported a better working alliance with the CA were more adherent to the intervention. This result is in line with studies about regular face-to-face interventions, digital therapy or treatment, and automated digital interventions. Nonetheless, we did not find an effect of human cues on the reported working alliance with the CA. This lack might also be due to the fact that TCAs are unable to use non-verbal communication. Building a relationship is an ambiguous process, which is more difficult to establish in a less rich text-based communication environment. Moreover, in studies that did find an improved working alliance with a CA either the interactions with the agent or the intervention itself were longer compared to those in our study. In other studies, although a high working alliance was reported within shorter periods of time, the interactions with the CA followed after introduction by a human healthcare professional. It is therefore unclear whether a TCA is less able to build a relationship with the user, or that it requires a longer time or introduction in a face-to-face introduction to do so. So even though our findings do support that working alliance is an important mechanism within interventions with TCAs, how to foster a relationship with a TCA still remains a question. Limitations and Suggestions for Future Work We aimed to test the effects of visual and relational cues in general, and whether a combination of visual and relational cues would lead to a higher adherence than visual or relational cues alone. However, this led to a combination of various visual and relational cues that were simultaneously manipulated. Future studies could dismantle these, and compare the effects of individual visual and relational cues with another to test which have the biggest influence on intervention adherence. Secondly, to mimic human behavior, we intentionally chose to apply subtle human cues to our CA. However, participants might not have processed the messages of the agent elaboratively enough to notice these subtle cues, resulting in a lack of effects. We suggest that future studies investigate whether stronger cues are needed in TCAs compared to ECAs to have similar effects. Additionally, these studies could investigate whether longer interactions do lead to an improved working alliance, and thus adherence, for interventions with a TCAs that elicit human cues. Finally, we did not inform our participants beforehand whether they were interacting with a computer or a human being. Therefore, the expectations of people might have varied, which might have affected our results. Future studies could keep these expectations constant by being transparent about the true nature of the automated agent. Another option would be to manipulate the description of the CA to more closely represent a human being or a computer, and ask participants about their expectations towards support by a human being or a computer, to test how these influence the effects of human cues within automated interventions. Our study was the first to test the effect of human cues within TCA-supported lifestyle interventions. Future studies could investigate the differences in applying human cues between TCAs and ECAs. Our results suggest that the lack of using non-verbal communication limited the capability to successfully apply relational cues. It would be interesting to test this hypothesis, and how to overcome the lack of non-verbal communication within TCAs. Conclusion We found that human cues do not improve adherence to TCA-supported interventions, and that visual cues even lead to lower levels of adherence. This is in contrast to the positive results of human cues found in studies with ECAs. In line with previous studies, we did find a positive relationship between working alliance with the TCA and adherence. The results suggest that being transparent about the computer-based nature of a CA and thereby setting the right expectations might be key. Besides, we found that factors that work for ECAs, in this case human cues, possibly cannot be carelessly copied to TCAs. This knowledge could help us gaining further knowledge that help us design better automated interventions in the future, which lead to better working alliances, higher levels of adherence, and in turn a healthier lifestyle for us all. University) for her assistance in the practical implementation of the study procedure, and during the development of the intervention. Conflicts of Interest None declared. Analyses with effectiveness as outcome variable Effectiveness was measured through objective step count data retrieved from Apple Health or Google Fit (depending on the smartphone of the participant). We calculated the mean difference between the average baseline step count (measured in the week before the intervention) and the average step count in the final week of the intervention. Participants were included in the analyses if both a valid baseline step count and a minimum of 5 days of step count in the final week were registered. We conducted an one-way between subjects ANOVA to compare intervention effectiveness between the four different conditions. There was no significant difference in effectiveness between the conditions, F =.726, P =.54 (see Table 3 for mean and SD per group). Also the post-hoc tests comparing the three human cues conditions with the no human cues condition (t = -1.021, P =.31), and the test comparing the condition with both visual and relational cues with the visual cues only and relational cues only groups (t =.171, P =.87) showed no significant differences.

Not all living situations are equal, and none are more aware of this than the residents of Maya and Olmeca, two of San Diego State’s freshman dorms. Neither hall has been renovated since their construction in 1960, which has led to a wide array of problems from regularly broken amenities to evacuation-forcing mold levels. Residing in any kind of dorm has its downsides, but conditions in Maya and Olmeca are unacceptable for the cost students pay. “It is unfair that Maya and Olmeca have the same mandatory price as the other nice dorms when they have zero advantages over them and haven’t been renovated, besides maybe a paint job, in so long,” said finance sophomore Teague Quillin who lived in Olmeca as a freshman. The price of living in any SDSU freshman residence halls is steep, at more than $1000 a month, even before factoring in extra required costs such as meal plans and ARC memberships. That’s just the price of living on campus. But, it is absurd that students living in the newly renovated South Campus Plaza and those living in Maya or Olmeca pay the same despite the substantial gap in housing quality. This gap in living conditions is appalling when one considers that the majority of freshmen are required by the university to live on campus. New students are subjected to lower living standards than they may be able to find off-campus for a cheaper price. SDSU offers cheaper rates for those who dorm with multiple roommates, so it should be possible for them to give discounts to students assigned to Maya or Olmeca as compensation for the potentially, and historically, unsanitary living conditions. Both buildings are scheduled to undergo remodelling this summer. This only kicks the problem further down the line. Given current construction trends, it looks as though all residence halls will be receiving a makeover at some point in the near future. But, there is no reason to believe that SDSU will not neglect another housing facility for another 50 years. It should have never gotten to this point. SDSU housing should take into account the problems that its residents face, and if such instances that have occurred in Maya and Olmeca occur in another hall one day, those students should be offered a discounted cost or at least the option to transfer to another building.

Norovirus foodborne outbreak in an agritourism: causes, development and legal consequences Norovirus is the main causative agent of viral acute gastroenteritis. It is easily transmitted and frequently causes foodborne and waterborne outbreaks. The Food Hygiene Service of ASL 2 Abruzzo received notification of gastrointestinal illness in 33 customers who attended a lunch at an agritourism on 28 July 2019. The investigation started immediately. The event was reported to the Judicial Authority and food preparation was suspended. Stool specimens and rectal swabs were collected from 18 guests and 2 food handlers and tested. A questionnaire survey was given to the customers. During the inspection, the Food Hygiene Service sampled some food leftover and water from faucets. Furthermore, it found an incomplete HACCP manual. The 31 cases of gastrointestinal illness occurred within 12-72hours and didn't need hospitalization. Among the affected subjects,18 were men (58.1%) with a median age of 51 years. All the biological samples collected from the subjects were positive for Norovirus GII, as revealed by Real Time-PCR. The two food handlers resulted positive for Norovirus GII, but they were asymptomatic. They were tested up to 3 weeks after the outbreak and their feces remained positive. Water and food samples resulted negative, except for raw ham's outer surface, that resulted positive for Norovirus GII. The contamination probably occurred during food preparation. This outbreak was related to food cross-contamination by infected food handlers, likely caused by a failure in application of both personal and food hygiene measures prescribed in HACCP plan. Food preparation restarted on 12 September 2019, but infected food handlers were dismissed until they were positive. The Judicial Authority will file a lawsuit against the food operator business that could lead to imprisonment or fines. Norovirus outbreaks are recognized as a public health concern causing a high number of cases and serious health consequences in children and the elderly. HACCP manual must be customized for each food business and integrally applied by food handlers in order to attain a higher standard of food safety for prevention of foodborne illness.

Chondroid metaplasia mimicking recurrent squamous cell carcinoma of the head and neck. A 65-year-old woman presented with a "sore bump" on her tongue. She had a history of squamous cell carcinoma of the head and neck that had been treated with surgery and radiotherapy 11 years earlier. The tongue lesion was excised, and pathologic examination identified a submucosal focus of benign-appearing cartilage. No evidence of dysplasia or malignancy was seen. She was diagnosed with chondroid metaplasia. Chondroid metaplasia involving the head and neck is rare. When it has occurred, it has been seen in both reactive and neoplastic settings. To our knowledge, this is the first reported case of a chondroid metaplasia of the tongue.

Star venture capitalist Ron Conway absolutely lost his shit today at the Bloomberg Next Big Thing Summit after fellow VC Chamath Palihapitiya suggested Mayor Ed Lee was doing a poor job addressing San Francisco's gentrification crisis. Conway, who raised $600,000 in angel funding for Lee's first mayoral campaign and has been an outspoken backer of the Mayor, had been stewing in the back of the auditorium for 10 minutes, according to TechCrunch, waving his hands demanding to comment on Palihapitiya's criticisms of Lee. And once Palihapitiya stated that "he'd resign" if he were Mayor Lee, Conway stood up to defend the Mayor, frantically pointing and shouting towards the speaker. "I live in the city of San Francisco, you live in the city of Palo Alto. [...] Ed Lee, who you ridicule—how dare you, Palo Alto resident!" Conway sneered as eyes widened behind him. Palihapitiya, an early employee of Facebook and investor in Yammer, attempted to have a conversation with Conway over housing policy. But Conway continued ranting, slamming Palihapitiya's apparent calls to levy a 1% equity tax on San Francisco startups to help fund affordable housing. "Ron, if [startups] wanted to be in the city, there are all these people in the city right now that are really frustrated. You see it with the riots. All I'm saying is..." "They're working to make it a better city, and so is Ed Lee, and it is going to get better—not worse." "Ron, effort is fine. All I'm saying is that there are a ton of people that feel like they're getting pushed out of subsidized housing, that feel like..." "30,000 units. Ed Lee has a mandate in every city department [Ed note: not true] to build 30,000 units. Is that not enough?" In fact, it isn't enough. According to study by San Francisco's chief economist Ted Egan, it would take at least 100,000 market rate units to make a noticeable dent in housing prices. "There's also the alternative of increasing market-rate housing construction, which does have an effect in the long run on housing prices," he said, adding that 100,000 new units — equal to all the housing built in San Francisco since the 1920s — is "the level of housing that you would have to build in order to see a significant increase in affordability at large." As for the mayor's recent plan to construct 30,000 new housing units, Egan said, "It's certainly fair to say 30,000 new units would be less impactful than 100,000, but it will have an impact." As much as Conway wants to point to Ed Lee's success in merely calling for 30,000 units to be built by 2020, the "follow through" is completely speculative. Mayor Lee's plan to cut red tape at the building permit office and bump up the priority of developments with high percentages of affordable housing is an ambitious approach towards fast tracking 30,000 units in a politically charged town. And never mind his office's very own inability to get already approved, city-funded affordable housing built in the city, which casts further doubt on his effectiveness. So it's unsurprising that Conway's blind faith in the Mayor wasn't enough to sway Palihapitiya. "All I'm saying is you can provide economic frameworks to solve these problems. You can say for all these companies that want to be [in San Francisco], you could [create] another kind of tax that says 'this is a subsidized housing tax' and we want a piece of your equity. And a company can chose not to be there. But imagine how many more units they could build? 300,000. A million!" Hell, 100,000 would be just fine. [Bloomberg]

Gel Spinning of Polyacrylonitrile/Cellulose Nanocrystal Composite Fibers. Polyacrylonitrile (PAN)/cellulose nanocrytal (CNC) fibers containing 0, 1, 5, and 10 wt % CNCs have been successfully produced by gel spinning. The rheological properties of solutions were investigated and the results showed that the complex viscosity and storage modulus of solutions were significantly affected by the presence of CNCs in the solution. Structure, morphology, mechanical properties and dynamic mechanical properties of these fibers have been investigated. Tensile modulus and strength increased from 14.5 to 19.6 GPa and from 624 to 709 MPa, respectively, as CNC loading increased from 0 to 10 wt %. Wide-angle X-ray diffraction results showed better PAN chain alignment and higher PAN crystallinity with the incorporation of CNCs.

A year ago, the Migration Advisory Committee (MAC) reviewed part of the country’s ‘shortage occupation list’ (SOL). The SOL features job titles and occupations with priority for Tier 2 work visas for skilled migrants from outside the European Economic Area (EEA). Migrants in such jobs do not have to earn the minimum £35,000 pay threshold that is now required to remain in the UK for longer than five years. The Department of Health did not initially request that nurses be put on the SOL. The MAC assumed, probably erroneously, that the department knows more about the UK labour market for nurses than the MAC does, so did not recommend that they be on the SOL. But during the course of 2015, the department altered its view, and to avoid putting the nation’s health at risk, the home secretary added nurses to the SOL in October pending a MAC review. An occupation must pass three hurdles to be placed on the SOL: is it skilled to the required level; is it in shortage; and is it sensible to fill vacancies with non-EEA labour? Here, I consider these questions, concluding with some observations about the nursing workforce that require urgent attention from the Department of Health and related bodies. Skilled, shortage and sensible? The Nursing and Midwifery Council, the professional regulatory body for nurses and midwives in the UK, is responsible for the registration of all UK nurses. It recognises four fields of registered nurse: adult; mental health; learning disabilities; and children. In England, there were 361,000 nurses working in the NHS in 2015. But in the UK as a whole – the NHS plus the care and independent health sectors – over 600,000 nurses are employed. The number of nurses has increased in the last three years. The OECD put the proportion of foreign-born nurses in the UK in total employment at 22 per cent in 2011, up from 15 per cent in 2001. Currently, the corresponding OECD nursing average is 15 per cent. In 2014/15, 8,000 foreign-born nurses were recruited to the UK, mainly from the EEA. The requisite skill level for inclusion on the SOL is National Qualifications Framework level 6 and above – that is, graduate level. Nurses are skilled to that level. Evidence from national data and partners indicates that nurses pass five of the seven MAC top-down indicators of shortage. These cover employment, hours worked and vacancies. There are also three pay indicators, but these are not relevant at a time of pay freeze or severe public sector pay restraint. Guidance from the National Institute for Health and Care Excellence indicates that organisations should aim for a maximum 5 per cent vacancy rate to accommodate operational flexibility needs. Health Education England, the body responsible for workforce planning for the NHS in England, estimates that the current vacancy rate for nurses in England is 9.4 per cent, nearly twice the recommended rate. In London, the Royal College of Nursing puts the rate at 17 per cent. Partner evidence suggests vacancy rates well above 5 per cent in the care sector too. Nurses’ pay accounts for about one tenth of NHS expenditure in England. In turn, spending on agency nurses is equivalent to one tenth of the nurses’ pay bill. Therefore, spending on agency nurses – around one per cent of NHS spending – should not be exaggerated. Nevertheless, such spending has risen rapidly in recent years, a further reflection of a shortage of nurses. National data and evidence from employers and trade unions therefore suggest a shortage of nurses. But why has this happened? Demand for nurses Four main factors have boosted the demand for nurses in recent years. The first three should surely have been anticipated by those responsible for workforce planning: Population: the total population is rising, people are living longer and they require more nursing care. Reforms: moves to integrate the NHS and social care, coupled with an emphasis on seven day working, raise demand. The changing role of nurses: nurses have taken on more responsibilities, including some duties previously carried out by doctors. If it is not used, the tension in policy objectives between restraining public spending and reducing immigration comes to the fore: The Francis report and staffing guidelines: demand for nurses rose as trusts sought to increase nurse-to-patient ratios following the 2013 Francis report into events at Mid Staffordshire NHS Foundation Trust. The supply side Supply is influenced by workforce planning, training places and retention efforts. Again, these are matters under the control of the Department of Health or individual employers: Workforce planning: in England, this involves aggregating local workforce plans into a national plan. The National Audit Office recently commented that this overlooks systemic changes in how services are delivered, and suggested that a more co-ordinated and proactive approach to managing the supply of staff could result in efficiencies for the NHS as a whole. Training: between 2009/10 and 2012/13, the volume of commissions (training places for nurses) fell by around a fifth (5,000 places). This trend has been partially reversed recently. The number of places would be substantially higher but for financial pressures. The move away from bursaries to a student loan system: in principle this is a sensible policy, but public sector pay restraint may limit the numbers prepared to take up the extra places providedby universities. Retention: the turnover of nurses rose from 7.8 per cent in 2008/09 to 9.3 per cent in 2014/15. There is now a noticeable spike in retirements at 55, the earliest age at which a nurse can retire on full NHS pension benefits. Considerable effort is being made to retain nurses. Local initiatives include flexible working, skills development and use of pay supplements. Pay Pay is a lever at the disposal of public sector employers to moderate shortages. If it is not used, the tension in policy objectives between restraining public spending and reducing immigration comes to the fore: Median pay for nurses is £31,500. This is £7,500 below median pay in other graduate occupations. There was a severe shortage of nurses in the late 1990s and early 2000s. The pay review body responded with substantial real pay increases. There is no sign of this happening now, nor of the Department of Health requesting such action. Available pay flexibility is insufficiently used. Possible adjustments include recruitment and retention premia and a market forces factor reflecting cost differences among healthcare providers. The MAC’s analysis shows that there is a historic pattern of peaks and troughs in the supply of migrant nurses. This pattern offers suggestive indications that migrant nurses have been used to save costs. Nursing is an occupation in which migrants earn, on average, less than UK workers doing the same job. In most other graduate occupations, migrants earn on average more than UK workers in the same job. It is difficult not to see this as undercutting. The evidence shows that non-EEA nurses are typically recruited at the minimum point on the nurses’ pay scale. More worryingly, pay at recruitment does not rise with age (an imperfect proxy for experience). A safety valve Over the next decade, the shortage of nurses can be addressed by more training places, reduced attrition among trainees, greater efforts at return to practice, more innovative use of pay flexibility and attention to working conditions. In the meantime, it is sensible to add nurses to the SOL. But there is a problem. The MAC was told by the Department of Health that employers in England will look to recruit around 11,000 non-EEA nurses over the next four years. But including nurses for Northern Ireland, Scotland and Wales, the actual figure could be over 14,000, approaching the annual quota of Tier 2 visas (20,700). Clearly, there is a danger that nurses – with their newly designated priority status – could crowd out skilled migrants from occupations not in shortage, including engineers and workers in the financial sector. To guard against this, the MAC recommends implementing a safety valve. We suggest an annual ceiling for nurses of 3-5,000 places in the first year. This might decrease year-on-year in line with the estimated required numbers set out by the Department of Health, such that nurses would come off the SOL in 2019 – the point at which the department forecasts that demand and supply of nurses will return to equilibrium. Challenges for the health and social care sector The MAC recommends placing nurses on the SOL: they are skilled, in shortage and – for a little while – it is sensible to add them. But we make this recommendation with considerable reluctance. It seems to us that the shortage is mostly down to factors that should have been anticipated by the Department of Health and related bodies. Furthermore, there seems to be an automatic presumption that non-EEA skilled migration provides the sector with a ‘get out of jail free’ card. Here, I briefly comment on just four areas: workforce planning; training commissions; pay; and who’s in charge. Workforce planning Until recently, workforce planning took no account of demand for nurses in the care and independent sectors, creating a structural undersupply of nurses in England. Similar issues apply in Scotland, Northern Ireland and Wales. Health Education England has now begun to factor demand for nurses from the care and independent sectors into their plans. Equally, the care and independent sectors make minimal effort to ensure that the number of nurses trained is sufficient to meet demand in their sectors. They make little or no direct contribution to the training of pre- registration nurses in the UK and seem content to have a free ride on the back of the government paying for training. Health Education England develops its workforce plans by adding together local workforce plans submitted by individual trusts. This means that systemic changes in demand – for example, the drive to integrate health and social care more effectively – are often not adequately reflected in the workforce plans. In addition, financial pressures in local trusts may lead them to understate their projected workforce needs. Training commissions The current shortage of nurses is closely linked to the decision to cut training places in England by more than 17 per cent between 2009/10 and 2012/13. The MAC has been told that this was driven more by financial issues than an expectation that demand would fall. Health Education England has recently confirmed that even now, the 331 additional places that it is funding in 2016/17 fall well below what is actually needed, again due to financial constraints. Pay The restraint on nurses’ pay instituted by the government was presented to us, and in the evidence to the pay review bodies, as an immutable fact. It’s not: it’s a choice. There was insufficient curiosity across both the health and care sector about the extent to which pay might be responsible for, and might help alleviate, present recruitment difficulties. By contrast, all parties seemed able to understand how their employees left for higher salaries available through agency work. Retention issues are a major contributor to current shortages in the NHS: the Department of Health should at least explore whether higher pay would improve retention. There is some evidence from the Institute for Fiscal Studies suggesting that nurses’ supply of labour to the NHS is sensitive to pay, most notably in London where the shortage appears to be particularly acute. Who’s in charge? There is a proliferation of bodies overseeing the administration of health and care services. The MAC received evidence from all of them but there was no common theme with a range of views expressed and data cited – and there is no single, authoritative voice to speak for them. We recognise the efforts of Health Education England to set up a group to pull together views on workforce planning, but the sectors do not help themselves by having a very confusing architecture. ♣♣♣ Notes: This article is based on the Migration Advisory Committee (MAC) report on nursing shortage. It appeared originally on CentrePiece, the magazine of LSE’s Centre for Economic Performance. The post gives the views of its authors, not the position of LSE Business Review or the London School of Economics. Featured image credit: DesignRaphael Ltd Before commenting, please read our Comment Policy David Metcalf is Emeritus Professor of Industrial Relations in LSE’s Department of Management and Associate in the Labour Markets Programme of the Centre for Economic Performance (CEP). Previously, he has been a visiting Professor at Princeton University and the Australian National University. He has worked extensively on the economic impact of unions and on pay systems and structures. Professor Metcalf is Chair of the Migration Advisory Committee, a Director of the Starting Price Regulatory Commission and a member of the Senior Salaries Review Body. Previously, he was a founding member of both the Low Pay Commission, which sets the national minimum wage, and the Union Modernisation Fund, a Government grant scheme providing financial assistance to independent trade unions and their federations. He owns a leg of a national hunt horse and is a Jockey Club Steward at Kempton Park and Plumpton. In 2008 he received a Commander of the Order of the British Empire (CBE) award for public service from Her Majesty the Queen.

Dr. Debra Shushan, APN Policy and Government Relations Director, was recently in the Philadelphia area where she was the featured speaker at a J Street Philadelphia / J Street U event at Haverford College and guest at parlor parties. In her September 30 talk at Haverford entitled "Off the Table: Israeli-Palestinian Peace in the Trump-Netanyahu Era," she surveyed recent developments that threaten prospects for peace - from the rise of illiberal democracy in the US and Israel, to President Trump's efforts to take key issues off the negotiating table - as well as the work that APN and Shalom Achshav in Israel are doing to push back against this tide. A J Street U leader called the talk "phenomenal" and welcomed further collaboration with APN. Educational advocacy is an important part of APN's work. If you are interested in inviting someone from APN's Speakers Bureau to your university, synagogue, church, parlor event, or other gathering, please contact apndc@peacenow.org.

Magnetite-supported hematin as a biomimetic of Horseradish peroxidase in phenol removal by polymerization Phenol removal using HRP and hematin as a biomimetic of HRP has been studied under various conditions at room temperature. The best results were obtained with treatment in two steps, with double addition of HRP or hematin and a final treatment with activated carbon. This two-step treatment achieved a minimum of 90% conversion of the initial phenol, under conditions commonly found in wastewaters (from 400 up to 1500 ppm phenol). Other additives such as chitosan, cellulose or polyethylene glycol (PEG) gave no satisfactory results. Hematin and magnetite-supported hematin showed comparable activities in phenol removal from aqueous solution. The supported hematin is an interesting alternative to HRP for practical application of a biomimetic catalyst for phenol removal.

Chronic Central Administration of Ghrelin Increases Bone Mass through a Mechanism Independent of Appetite Regulation Leptin plays a critical role in the central regulation of bone mass. Ghrelin counteracts leptin. In this study, we investigated the effect of chronic intracerebroventricular administration of ghrelin on bone mass in Sprague-Dawley rats (1.5 g/day for 21 days). Rats were divided into control, ghrelin ad libitum-fed (ghrelin ad lib-fed), and ghrelin pair-fed groups. Ghrelin intracerebroventricular infusion significantly increased body weight in ghrelin ad lib-fed rats but not in ghrelin pair-fed rats, as compared with control rats. Chronic intracerebroventricular ghrelin infusion significantly increased bone mass in the ghrelin pair-fed group compared with control as indicated by increased bone volume percentage, trabecular thickness, trabecular number and volumetric bone mineral density in tibia trabecular bone. There was no significant difference in trabecular bone mass between the control group and the ghrelin ad-lib fed group. Chronic intracerebroventricular ghrelin infusion significantly increased the mineral apposition rate in the ghrelin pair-fed group as compared with control. In conclusion, chronic central administration of ghrelin increases bone mass through a mechanism that is independent of body weight, suggesting that ghrelin may have a bone anabolic effect through the central nervous system. Introduction The brain is regarded as the master regulator of homeostasis and metabolism. It has been suggested that bone and energy metabolism require tightly coordinated regulation so that longitudinal growth, or bone remodeling, are in accordance with energy supply and demand. Numerous studies have investigated the mechanisms involved in the central regulation of bone metabolism and possible connections between bone metabolism and energy metabolism. However, the data are contradictory regarding bone metabolism regulation. In addition, the factors that co-regulate bone metabolism and energy metabolism are not yet clear. Leptin, a major adipokine that regulates appetite, has been widely investigated as the major factor co-regulating bone metabolism and energy metabolism. Ducy et al. demonstrated that intracerebroventricular (ICV) injection of leptin induced bone loss in the spine, suggesting a role for leptin in bone metabolism regulation through a central mechanism. Data from subsequent reports supported the bone catabolic effect of leptin and have shown that leptin inhibits bone formation via the sympathetic nervous system. However, several recent studies have reported contradicting data. Leptin administration via gene therapy into the hypothalamus did not have a significant effect on bone metabolism. Leptin receptor deficient mice exhibit decreased bone mass, demonstrating leptin's anabolic effect on bone. Furthermore, ICV injection of leptin increases bone mineral density (BMD) and mineral apposition rates, a finding which is the exact opposite of the initial studies reporting a catabolic effect of central leptin. Given this conflicting data, further studies are required to clarify the mechanisms underlying the central regulation of bone metabolism. Ghrelin is a stomach hormone that acts centrally to increase appetite. Several studies have investigated the effect of chronic ICV ghrelin on energy metabolism. Chronic ICV ghrelin infusion increased food intake and weight gain, reversed the effect of leptin on food intake, increased the glucose utilization rate of adipose tissue, and increased lipogenic enzymatic activity in adipose tissue and liver. A recent study investigated the central effects of ghrelin and leptin on body and bone marrow adiposity, as well as adipose tissue and bone marrow gene expression, and reported central ghrelin had no effect on bone marrow adiposity. However, to date, no study has investigated the effect of chronic ICV ghrelin on bone metabolism. Ghrelin and leptin have opposite effects on energy metabolism, but also on the sympathetic nervous system and other pathways. Central leptin stimulates sympathetic outflow whereas central ghrelin suppresses sympathetic activity. Since the sympathetic nervous system has been suggested as the major pathway governing the effect of central leptin on bone metabolism, it is possible that sympathetic suppression through central ghrelin can affect bone metabolism. Therefore, the present study was designed to investigate the chronic effects of centrally administered ghrelin on bone metabolism. Animals and peptide Male Sprague-Dawley rats (6 weeks old) weighing 180-230 g were used. Body weight and food intake were monitored daily. All animal experiments were performed with approval from the Institutional Animal Care and Use Committee (IACUC) of the Clinical Research Institute at Seoul National University Hospital (an AAALAC accredited facility). National Research Council (NRC) guidelines for the care and use of laboratory animals were also observed (1996 revision). The standard rodent chow (Purina Rodent Chow; Biopia, Korea) was used. Ghrelin (rat) was purchased from Bachem Inc. (Bubendorf, Switzerland).The ghrelin peptide was prepared with concentration of of 0.25 mg/ ml which corresponds to 1.5 mg/day (6 mg/day; 7.14 mg/Kg body weight/day). Surgery Rats were anesthetized by intraperitoneal injection of 50 mg/kg zoletil and 10 mg/kg xylazine and surgically implanted with a 22gauge stainless-steel cannula (Plastics One Inc. Roanoke, VA, USA) into the third cerebral ventricle. Osmotic mini-pumps (Model 2004, 0.25 mL/h; Alzet Corp., Cupertino, CA, USA) filled with saline or rat ghrelin peptide were implanted under the dorsal chest skin. The mini-pumps were connected to the ICV cannula via a catheter. Alzet Brain infusion kit 2 with infusion cannula of ID = 0.18 mm and OD = 0.36 mm was used. The cannula was stereotactically placed 0.72 mm posterior to the bregma on the midline and implanted 7 mm below the outer surface of the skull surgery via a stereotaxic apparatus. For the verification for correct cannula placement, cresyl violet staining and brain dissection was performed. Study design Fifteen rats were divided into 3 groups: the control group (n = 5) received ICV infusions of saline for 21 days; the ghrelin ad libitumfed (ghrelin ad lib-fed) group (n = 4) received ICV infusions of ghrelin (1.5 mg/day) for 21 days; the ghrelin pair-fed group (n = 6) received ICV infusions of ghrelin (1.5 mg/day) for 21 days and were allowed to eat only as much chow as consumed by the control group on the previous day. Radiological analyses and bone histomorphometry Nondestructive, three-dimensional evaluation of bone mass and architecture were performed using a microCT scanner (Skyscan 1076 for tibia and lumbar spine and Skyscan 1172 for femur; Skyscan, Aartselaar, Belgium). Lumbar spine and femur were dissected from soft tissue, fixed in 70% ethanol, and analyzed. Image acquisition of tibia and lumbar spine L3 was performed with a source voltage of 100 kV, current of 100 mA, a 0.5-mm aluminum filter, and an isotropic voxel size of 8.8 mm. Image acquisition of femur was performed with a source voltage of 70 kV, current of 141 mA, a 0.5-mm aluminum filter, and an isotropic voxel size of 11.55 mm. For the tibia metaphysis trabecular bone, 251-slice-thick volume of interest starting 150 slices distal to the proximal growth plate were analyzed. For the tibia diaphysis cortex, the mid-diaphysis cortical bone volume of interest was analyzed. For the femoral metaphysis, 101-slice-thick volumes of interest starting 150 slices proximal to the distal growth plate were analyzed. For the lumbar spine, trabecular bone volumes of interest of whole trans-axial spine body images were analyzed. For trabecular volumetric bone mineral density (BMD) analyses using microCT, phantoms with predefined densities and CTA software were used for BMD measurements, as described in the manufacturer's manual (Skyscan, Aartselaar, Belgium). The trabecular and cortical bone volumes of interest were outlined by interpolation of operator-drawn regions exclusively representing the trabecular and cortical bone, respectively. BMD (g/cm 2 ) of the ex vivo tibia, femur, and lumbar spine (L2 and L3) were measured using the dual energy X-ray absorptiometry (DXA) instrument PIXIMUS (GE Lunar, Madison, WI, USA). A phantom was scanned daily for quality control. For histomorphometric analyses of dynamic parameters, rats were injected with calcein (20 mg/kg body weight) 2 and 6 days prior to sacrifice. Dynamic histomorphometry analyses were conducted on the lumbar spine L5 using the Bioquant program (Bio-Quant. Inc., San Diego, CA, USA). Statistical Analyses Data are presented as the mean 6 SEM. Statistical analyses were performed using analysis of variance (ANOVA) with least significant difference (LSD) as a post hoc comparison. Significance was defined as P,0.05. Statistical analyses were performed with SPSS for Windows (version 17.0, SPSS Inc., Chicago, IL, USA). Chronic ICV ghrelin infusion increases body weight and food intake Chronic ICV ghrelin infusion (1.5 mg/day for 21 days) significantly increased body weight in ghrelin ad lib-fed compared with ICV saline-infused control rats (33164 g vs. 31269 g, P,0.05). However, there was no difference in body weight between the control rats and ICV ghrelin-infused rats that were pair-fed the food intake of the control rats (ghrelin pair-fed) (316611 g) (ANOVA F value = 4.21) (Fig. 1A). Although there was no significant difference in cumulative food intake between the ghrelin pair-fed and control groups, ghrelin ad lib-fed rats had significantly higher cumulative food intake compared with ghrelin pair-fed rats (P,0.05) (ANOVA F value = 2.86) (Fig. 1B). Chronic ICV ghrelin infusion increases bone mass Chronic ICV ghrelin infusion significantly increased bone mass in ghrelin pair-fed group compared with control rats. This was indicated by increased bone volume percentage (BV/TV, 32.461.5% vs. 24 (Fig. 2). The trabecular pattern factor (Tb.Pf), a parameter inversely correlated to the connectivity, was significantly decreased in the ghrelin pairfed group, indicating that the trabecular structure was more connected in the ghrelin pair-fed group (P,0.05, Fig 2) (ANOVA F value = 6.46). This result is consistent with the decreased structure model index (SMI) observed in the ghrelin pair-fed group, which suggests that the trabecular structure was more plate-like in the ghrelin pair-fed group, as compared with the rodlike structures in the control group (P,0.05, Fig. 2) (ANOVA F value = 4.45). There were no significant differences in bone mass observed between the control group and ghrelin ad-lib fed group. Similar trends were observed in femur and lumbar spine, with lower statistical significance ( Figure S1 and S2). Cortical area (Ct.Ar) was significantly increased in the ghrelin ad lib-fed group compared to the control group (P,0.05, Figure S3) (ANOVA F value = 3.57). There was a tendency of increased cortical area fraction and cortical thickness in the ghrelin ad lib-fed group compared to the control group (P,0.1, Figure S3) (ANOVA F value = 1.73 and 2.49, respectively). There were no significant differences in ex vivo BMD measurements between the three groups ( Table 1). Chronic ICV ghrelin infusion increases the mineral apposition rate Chronic ICV ghrelin infusion significantly increased the mineral apposition rate (MAR) in the ghrelin pair-fed group compared with the control group (5.060.2 mm/d vs. 4.060.1 mm/d, P = 0.014, Fig. 3) (ANOVA F value = 5.36). The ghrelin ad libfed group tended to have a higher MAR compared with the control group (P = 0.073, Fig 3). No significant differences in bone formation rate were observed among the three groups. Effect of chronic ICV ghrelin infusion on serum markers Chronic ICV ghrelin infusion significantly increased serum leptin in both the ghrelin ad lib-fed group and ghrelin pair-fed group compared with the control group (Fig. 4). Chronic ICV ghrelin infusion significantly decreased serum ghrelin in the ghrelin ad lib-fed group compared with the control group (P,0.05) (ANOVA F value = 3.55 and 6.31, respectively). However, there were no significant differences in IGF-1, CTX, TRAP-5b, and P1NP among the three groups. Discussion Using a rat ICV administration model, we demonstrate that intracerebral infusion of ghrelin results in increased bone mass, connectivity, and mineral apposition rates in ghrelin pair-fed group. Chronic ICV administration of ghrelin increased weight gain in the ghrelin ad lib-fed group; however, this effect was abolished by pair feeding. These findings indicate that chronic central administration of ghrelin increased bone mass through mechanisms independent of changes in body weight. The duration of ICV ghrelin treatment (21 days) in this study could be considered relatively short to induce profound changes in bone mass. However, it should be noted that establishment of an animal model of chronic ICV treatment is technically challenging and the present study is the longest ICV ghrelin treatment attempted (21 days), with previous studies infusing ghrelin for only 6-12 days 12]. Future study using Alzet osmotic pump model 2ML4 could extend chronic infusion up to 28 days. Although we clearly demonstrated anabolic effects of ghrelin on bone (independent from ghrelin-induced effects on feeding), the mechanism for this effect is not known. One possible mechanism may be suppression of the sympathetic nervous system. Sympathetic activity has been reported to be suppressed by ICV administration of ghrelin and stimulated by ICV administration of leptin. Hypothalamic administration of leptin was reported to decrease bone formation due to the ability of leptin to increase sympathetic tone. Numerous animal and human studies demonstrate that beta-blockers have protective effects on bone metabolism. Therefore, ICV ghrelin may increase bone mass through sympathetic suppression. Future studies using pharmacologic blockade of the sympathetic nervous system or local sympathectomy surgery will clarify this mechanism. Other possible mechanisms include the secretagogue effect of ICV ghrelin on growth hormone. However, the control group and ghrelin pair-fed group did not differ in serum IGF-1 levels so this does not support a role for growth hormone. These data are supported by a previous study that also reported no change in plasma IGF-1 after 7 days of ICV ghrelin treatment. It has also been reported that ICV administration of ghrelin increased the plasma concentration of growth hormone on day 6 but this was not sustained on day 12. Therefore, it is unlikely that the effect of ICV ghrelin is mediated through a growth hormone-IGF-1 axis. In contrast to the ghrelin pair-fed group, the ghrelin ad lib-fed group had a slight increase in cortical bone area and no significant difference in trabecular phenotypes compared with controls. However, the ghrelin ad lib-fed group did exhibit increased body weight gain. Therefore, the increased cortical bone mass could be the result of increased body weight and a consequent increase in mechanical loading on bone. Recently, we and others have reported that increased fat mass could have adverse effects on bone mass because of deleterious metabolic effects. In addition, many researchers have suggested that weight gain and obesity could increase cortical bone mass through mechanical loading with a resultant decrease in trabecular bone due to metabolism or systemic effects. Fatty acid lipotoxicity, numerous adipokines, inflammatory cytokines, aromatase, and insulin resistance have been suggested to mediate the deleterious effects of fat on bone metabolism. Therefore, the absence of an increase in trabecular bone mass observed in the ghrelin ad lib-fed group could in part result from the deleterious effects of increased fat mass. The ghrelin ad lib-fed group had a significantly decreased serum ghrelin concentration, likely due to a compensatory response to increased body weight. And this decrease in serum ghrelin could result in decreased bone formation, which would then neutralize the anabolic effect of ICV ghrelin on bone. Central ghrelin is reported to have an effect on adipose tissue independent from its effects on food intake. Therefore, the ghrelin pair-fed rats could have some changes in adipose tissue deposition, which could be independent of food intake and weight change. These changes in adipose tissue deposition could have some direct and indirect effect on the bone phenotype of these ghrelin pair-fed rats. Furthermore, the ghrelin effect on adipose tissue deposition may have contributed to the lack of weight gain in the ghrelin pair-fed rats. However, we did not investigate the adipose tissue phenotypes in the present study to determine the role of adipose tissue deposition in the central regulation of bone metabolism. In contrast to the increased bone mass determined by microCT measurement, there were no significant changes in ex vivo BMD measured by DXA in tibia, femur, and lumbar spine among the three groups. Several possible explanations may account for the discrepancy. DXA measurement reflects both trabecular and cortical bone mass, whereas microCT gives separate estimates of BMD for trabecular and cortical bone as well as reports volumetric mineral density in g/cm 3. Therefore, DXA measurements cannot identify the difference in trabecular bone observed by microCT. It has been reported that the measurement precision of the excised femur BMD is generally not as precise as that for the intact femur BMD in vivo, although the general efficacy of ex vivo DXA measurements were acceptable. Although similar trends were observed, the effects of ICV ghrelin on femur and spine were not statistically significant. There could be a skeletal site-specific effect of ICV ghrelin. Interestingly, a previous study reported a skeletal site-specific effect of leptin with lower femur BMD and higher spine BMD observed in leptin deficient mice. However, no skeletal site-specific effect was observed in leptin receptor-deficient mice. We measured serum ghrelin to investigate possible leakage of ghrelin into systemic circulation from the ICV injection. However, we found a significant decrease in serum ghrelin in the ghrelin ad lib-fed group as compared to the control group. This result indicated that leakage to systemic circulation was unlikely. This decrease in serum ghrelin and increase in serum leptin could be a compensatory response to the weight gain of the ghrelin ad lib-fed group. However, the increase in serum leptin in ghrelin pair-fed group is not due to the weight gain, since there was no weight gain in the ghrelin pair-fed group. Another possible explanation is that the increase in serum leptin observed in the ghrelin ad lib-fed group and ghrelin pair-fed group is likely a compensatory response related to ICV ghrelin injection. Previous studies have reported a similar trend of increased blood leptin levels in ICV ghrelin injected animals. Circulating ghrelin and leptin may contribute to bone metabolism via a direct effect on bone cells. Generally, ghrelin increases both osteoblast and osteoclast function and leptin increases osteoblast function but decreases osteoclast function. However, these direct effects of ghrelin and leptin were inconsistent depending on the concentrations, assays, and cell types used. Therefore, the role of serum ghrelin on bone metabolism is complex, and future study specifically aimed to investigate the role of serum ghrelin on bone metabolism is required to determine this issue. We observed no significant differences in CTX, TRAP-5b, or P1NP, which are serum biochemical markers of bone turnover. Since ghrelin was placed in a subcutaneously transplanted osmotic pump, it is possible that after 21 days of incubation at body temperature, the effect of ghrelin was diminished due to peptide degradation. Therefore, the serum biochemical markers measured after 21 days of treatment may not have captured the dynamic bone metabolism state during the treatment period. However, other possibilities could be limitations from the ELISA assay or serum preparation procedure. The findings of the current study have important clinical implications since ghrelin or ghrelin mimetics could be utilized as potential therapeutic modalities for osteoporosis. A cross-sectional study showed that serum ghrelin positively correlated with BMD. In addition, there are a number of clinical trials investigating the effects of ghrelin or ghrelin mimetics on various conditions, including sarcopenia, cancer-related cachexia, anorexia nervosa, cystic fibrosis, postoperative gastric ileus, and gastroparesis. These studies have already validated the safety and efficacy of ghrelin or ghrelin mimetics. First limitation in the present study is the dose of ICV ghrelin treatment (1.5 mg/day). This dose is slightly higher than some studies (1.0 mg/day and 1.2 mg/day). However, other studies have used higher doses (5-20 mg/day). The dose of present study (1.5 mg/day) could be insufficient or subthreshold to induce adequate central ghrelin effect. Second limitation was the statistical power of the present study. Most of the findings are based on LSD post hoc comparisons, which is very liberal. Additional post hoc comparisons with Scheffe and Tukey HSD test were performed and some of the findings (trabecular number, cortical bone area, serum ghrelin, body weight and food intake) did not reach statistical significance. And, it should also be noted that some of the findings with tendency of P,0.1 may be false positives. Third limitation was the conflicting fact that long duration of treatment was required to induce a sufficient bone metabolism change, whereas long duration of incubation in body temperature result in risk of ghrelin peptide degradation. In conclusion, chronic central administration of ghrelin increases bone mass through a mechanism that is independent of body weight, suggesting that ghrelin may have a bone anabolic effect through the central nervous system.

Conrad Santos Conrado de Regla Santos (November 26, 1934 – February 29, 2016) was a politician in the province of Manitoba, Canada. He was a member of the Legislative Assembly of Manitoba from 1981 to 1988, and again from 1990 to 2007. Conrado as his family and friends called him, had three children: Evelyn Santos, Conrad Santos and Robert Santos. The son of Federico Santos and Marcelina de Regla, he was born in the Philippines and was educated at Harvard University and the University of Michigan, receiving a PhD in Political Science from the latter institution. He moved to Winnipeg, Manitoba, Canada in 1965 after receiving a teaching position at the University of Manitoba, where he retired from in 2008. Santos has also worked as a consultant for the Instituto Centro-Americano de Administracion Publica in Costa Rica, and was a board member of the Citizenship Council of Manitoba from 1977 to 1980. He sought the New Democratic Party of Manitoba (NDP) nomination for Fort Garry in the 1973 election, but was defeated. He ran for the Winnipeg city council in 1977 and 1980, but lost both times. Santos was first elected to the Manitoba legislature in the 1981 provincial election as a New Democrat in the northwest Winnipeg riding of Burrows, defeating NDP-turned-Progressive Member of the Legislative Assembly (MLA) Ben Hanuschak. He was re-elected in the 1986 election. In June 1984, there were unconfirmed rumours that he was considering a move to the Progressive Conservative Party. Santos lost the Burrows NDP nomination to Doug Martindale in 1988, and subsequently entered the party's leadership election. He was not regarded as a prominent candidate, and received only five votes on the first ballot. He ran for mayor of Winnipeg in 1989, and finished a distant fourth. In 1990, Santos won the NDP nomination for Broadway by a single vote over Marianne Cerilli, whose candidacy was supported by the party leadership. He defeated Liberal incumbent Avis Gray in the 1990 general election, and was re-elected in the 1995 election. In 1995, he endorsed Lorne Nystrom's bid to lead the federal NDP. When the Broadway riding was eliminated by redistribution in 1999, Santos won the NDP nomination in Wellington (also in Winnipeg's northwest), and was returned by a wide margin in the 1999 provincial election. He was again re-elected in the 2003 election. Santos was named Deputy Speaker after the elections of 1986 and 1999. Santos left the New Democratic Party caucus shortly before the 2007 provincial election after being accused of improperly selling party membership cards. He campaigned as an independent, and finished last in a field of five candidates. He later pleaded guilty to paying the membership fees of as many as one hundred new party members, during the time when he was trying to retain his nomination. He was fined $200 under the provincial Election Finances Act, as well as court fees of $150. His lawyer argued that Santos was simply trying to help his low-income supporters, saying that the MLA "didn't have the heart" to request membership fees from people who were unable to feed their families. He married Emerita Maglaya. Santos died in Winnipeg on February 29, 2016 at the age of 81.

A telephone hotline for transient ischaemic attack and stroke: prospective audit of a model to improve rapid access to specialist stroke care PROBLEM Patients with transient ischaemic attack or stroke benefit from early diagnosis, specialist assessment, and treatment with thrombolysis, and from stroke unit care and secondary prevention. The challenge with such patients is to minimise delays and ensure that treatment is appropriate, and to provide this care with the available resources. DESIGN An ongoing prospective audit of a transient ischaemic attack and stroke clinic (1 January 2005 to 30 September 2009), as part of the Scottish Stroke Care Audit, and a three month targeted audit of immediate telephone access to a specialist stroke consultant (1 February 2009 to 30 April 2009). SETTING Stroke and transient ischaemic attack services in Lothian, a region of Scotland with a population of 810,000. KEY MEASURES FOR IMPROVEMENT Delays to assessment at a rapid access transient ischaemic attack and stroke clinic; delays to appropriate treatment. STRATEGY FOR CHANGE In February 2007 we introduced a 24 hours a day, seven days a week hotline to a consultant, who provided immediate advice on diagnosis, investigation, and emergency treatment for patients with transient ischaemic attack or stroke, and suggested the most appropriate care pathway, which might include an early appointment in a transient ischaemic attack and stroke clinic. EFFECTS OF CHANGE The introduction of the hotline was associated with an immediate and sustained reduction in delays to assessment (from 13 to three days) and treatment. The proportion of participants taking statins at the time of visiting the clinic increased from 40% before the introduction of the hotline to 60% after the hotline was in place. Also, the hotline contributed to a reduction in the delay from last event to carotid surgery, from 58 days to 21.5 days. A total of 376 calls were received during the three month audit. Of the 273 (88%) referrers who responded to our questionnaire, 257 (94%) were very satisfied with the advice given over the hotline. LESSONS LEARNT Although associated with some disruption to the activities of the consultants, a 24 hours a day, seven days a week telephone hotline to a consultant is a feasible and effective means of reducing delays to specialist assessment and treatment of patients with transient ischaemic attack or stroke.

A Review of Irradiation Damage and Effects in -Uranium Understanding irradiation damage and effects in -uranium (-U) is critical to modeling the behavior of U-based metallic fuels. The aim of this review is to address the renewed interest in U-based metallic fuels by examining the state-of-the-art knowledge associated with the effect of irradiation on the microstructure, dimensional changes, and properties of -U. We critically review the research progress on irradiation-induced growth and swelling, the enhancement of plastic flow and superplasticity by irradiation, and the effect of irradiation on thermal and electrical properties of -U. Finally, we outline the research directions that require advancements, specifically the need to carry out fundamental research on several of the less understood mechanisms of irradiation damage and effects in -U. Introduction Although the current light water reactors predominantly use uranium dioxide as the major fuel elements, metallic uranium and uranium alloys were extensively considered for fast reactor applications in 1950s and 60s due to their higher fissile and fertile material density compared to oxides. Recently, the nuclear engineering community has renewed interest in several metallic uranium concepts for use in Generation IV reactor designs. Under the Department of Energy, the advanced fuels campaign is focusing on metallic uranium concepts for sodium fast reactors. -U is an important allotrope of solid uranium observed at low temperatures (<940 K). Much of the research on -uranium occurred after World War II to solve the persistent problems encountered in preparing uranium metal for a nuclear fuel and for plutonium production (particularly at Hanford and Savannah River sites). There was a large effort in the late 1940s and early 1950s, particularly at Argonne National laboratory and Knolls Atomic Power Laboratory, to develop an understanding of the anisotropic thermal expansion, irradiation-induced swelling and irradiation creep of -uranium. Some of the comprehensive reviews on the properties of metallic uranium with a focus on irradiation effects were reported in late 1950s and 60s. -U exhibits several unusual properties, including high-pressure superconductivity, high plastic flow induced by irradiation growth, and massive, catastrophic irradiation swelling (referred to as "breakaway swelling") at a temperature of around 923 K. Several of these properties are not completely understood even today. However, the fundamental research on irradiation-induced dimensional growth of -U progressed into the late 1960s and early 1970s, which also included a worldwide effort in measuring its physical properties (such as electrical resistivity and thermal conductivity) and mechanical properties (such as plastic deformation). The nuclear industry, however, gradually moved forward with ceramic UO 2 fuels, leading to a rapid decline in metallic uranium research. However, the condensed matter physics community had continued interest in -U as a superconductor. In 1994, Lander et al. comprehensively covered the properties of unirradiated uranium, primarily focusing on its electronic and mechanical properties. However, -U has witnessed uranium, primarily focusing on its electronic and mechanical properties. However, -U has witnessed renewed interest in the nuclear industry in the last decade, primarily because it is a component of certain metallic fuels for fast reactors. The present review focuses on the fundamental insights available from the research over the last few decades, with focus on the recent research progress on irradiation damage and effects in -U. This review starts by tabulating the crystal structure and mechanical and physical properties of -U in Section 2. In Section 3, irradiation-induced microstructural and dimensional changes in -U are reviewed. Following this, we summarize the effect of irradiation on its mechanical and physical properties. Finally, in Section 4, we provide a brief outlook for future research. Crystallography and Fundamental Defect Kinetics The phase of uranium consists of corrugated sheets of atoms in a highly asymmetrical face-centered orthorhombic structure. Beeler et al. calculated the lattice constants of -U with a = 2.793, b = 5.849, c = 4.894, as seen in Figure 1a. The structure of -U is made up of close-packed corrugated planes of atoms, including planes parallel to the densely packed flat plane and planes parallel to the axis. The interatomic distances of corrugated planes (e.g., line segments ab = 2.70 and ad = 2.80 in Figure 1b) are smaller than interatomic distances between the corrugated planes (e.g., line segment ak = 3.25 and dk = 3.34 in Figure 1b). This corrugated structure is responsible for the anisotropy of physical properties of -U. There is only one type of lattice site, tetrahedral interstitial site, pentahedron site, and single vacancy site. However, there exist several types of self-interstitial configurations (eight tetrahedral, eight pentahedral pyramidal and four octahedral interstitial sites). The most stable configuration is schematically shown in Figure 1b, where atom X is in the interstice of a pentahedron., 2022, Elsevier). (b) schematic structure of -U and the most stable self-interstitial configuration (reprinted and adapted with permission from, 2022, IOP publishing, Ltd). Recent calculations of point defect formation and displacement energies indicate that radiation damage is expected to be less significant in the -phase compared to the -phase. The threshold displacement energy of -U was estimated using density-functional theory (DFT) and molecular dynamics (MD) calculations. The threshold displacement energy of -U was estimated to be 63.4 eV at 800 K. Compared to 35.6 eV in -U at 800 K, more energy is required to displace an atom from a lattice site in -U and the number of collision cascades from the same primary knock-on energy is reduced. The minimum threshold displacement energy (~20 eV) exists along the to directions and hence these are expected be directions of easier defect creation. Beeler et al. also reported the defect formation energies and diffusion coefficients in -U at 600 K and 800 K through ab, 2022, Elsevier). (b) schematic structure of -U and the most stable self-interstitial configuration (reprinted and adapted with permission from, 2022, IOP publishing, Ltd.). Recent calculations of point defect formation and displacement energies indicate that radiation damage is expected to be less significant in the -phase compared to the -phase. The threshold displacement energy of -U was estimated using density-functional theory (DFT) and molecular dynamics (MD) calculations. The threshold displacement energy of -U was estimated to be 63.4 eV at 800 K. Compared to 35.6 eV in -U at 800 K, more energy is required to displace an atom from a lattice site in -U and the number of collision cascades from the same primary knock-on energy is reduced. The minimum threshold displacement energy (~20 eV) exists along the to directions and hence these are expected be directions of easier defect creation. Beeler et al. also reported the defect formation energies and diffusion coefficients in -U at 600 K and 800 K through ab initio MD simulations. The estimated interstitial and vacancy diffusion coefficient in the 600-800 K range was 2.172 10 −7 and 1.148 10 −7 m 2 /s, respectively. More recently, in the variation in defect formation energy with temperature was studied via ab initio MD and is shown in Figure 2. The defect formation energy for both vacancy and interstitial defects tends to slightly decrease until 400 K and then increases with increasing temperature. initio MD simulations. The estimated interstitial and vacancy diffusion coefficient in the 600-800 K range was 2.172 10 −7 and 1.148 10 −7 m 2 /s, respectively. More recently, in the variation in defect formation energy with temperature was studied via ab initio MD and is shown in Figure 2. The defect formation energy for both vacancy and interstitial defects tends to slightly decrease until 400 K and then increases with increasing temperature. Electronic Properties The electronic structure of uranium is dominated by 5f electron states that form narrow bands that tend to bond the atoms in complex and distorted ways. DFT results reveal that 5f orbitals are active in the chemical bonding for uranium. The behavior of 5f electrons of uranium is observed to be similar to that of the d orbital electrons in the transition elements. Compared with the bulk atoms, the deficit of atoms near the uranium surface breaks the balance between the delocalization and localization of 5f electrons, leading to the electrons changing from delocalized to localized. As the electrons change from delocalized to localized, the arrangement of the surface atoms is interrupted and the total energy of the surface increases. This principally leads to a high surface energy, in turn leading to a high affinity for hydrogen, oxygen and carbon. The research on the electronic properties of -U was mainly motivated by the anomalous electronic configuration of -U and the discovery of superconductivity in -U. A long-standing debate on the description of the 5f elements involved whether they should be called actinides, in which the 5f shell was progressively filled, or thorides, in which the 6d shell progressively filled. From the chemical point of view, this question was effectively settled by the production of the heavier elements Np and Pu and the early study of their spectroscopic properties, revealing the actinide nature of -U. Superconductivity was discovered in -U in 1942 by Aschermann and Justin and was first reported by Mott. However, the existence of intrinsic superconductivity in -U (i.e., superconductivity of -U in the pure undoped state) is a topic of discussion even today. Multiple investigations in the 1950s found a range of superconductivity transition temperatures Electronic Properties The electronic structure of uranium is dominated by 5f electron states that form narrow bands that tend to bond the atoms in complex and distorted ways. DFT results reveal that 5f orbitals are active in the chemical bonding for uranium. The behavior of 5f electrons of uranium is observed to be similar to that of the d orbital electrons in the transition elements. Compared with the bulk atoms, the deficit of atoms near the uranium surface breaks the balance between the delocalization and localization of 5f electrons, leading to the electrons changing from delocalized to localized. As the electrons change from delocalized to localized, the arrangement of the surface atoms is interrupted and the total energy of the surface increases. This principally leads to a high surface energy, in turn leading to a high affinity for hydrogen, oxygen and carbon. The research on the electronic properties of -U was mainly motivated by the anomalous electronic configuration of -U and the discovery of superconductivity in -U. A long-standing debate on the description of the 5f elements involved whether they should be called actinides, in which the 5f shell was progressively filled, or thorides, in which the 6d shell progressively filled. From the chemical point of view, this question was effectively settled by the production of the heavier elements Np and Pu and the early study of their spectroscopic properties, revealing the actinide nature of -U. Superconductivity was discovered in -U in 1942 by Aschermann and Justin and was first reported by Mott. However, the existence of intrinsic superconductivity in -U (i.e., superconductivity of -U in the pure undoped state) is a topic of discussion even today. Multiple investigations in the 1950s found a range of superconductivity transition temperatures depending on the purity of the samples. Conversely, several late 1960s studies indicated the possibility of superconductivity being the result of grain-boundary filamentary sheathes rather than bulk superconductivity, indicating that superconductivity is not intrinsic to -U. In fact, Geballe et al. suggested that the variations in superconductive properties were correlated to the intergranular strains resulting from anisotropic expansion in -U. Three phase transitions occur in -U below 43 K due to the presence of charge density waves and were postulated to affect the intrinsic superconductivity of -U. Charge density waves (CDW) occur when there is a spatial modulation of the electron density. This CDW is typically associated with a mass density wave of the positively charged ions which make up the crystal lattice. The possibility of CDW formation is enhanced in lowerdimensional systems (1D and 2D materials) because the simple structures involved lead to a high probability of favorable Fermi surface nesting (i.e., the shifting and coincidence of two Fermi surfaces that are required for CDW formation). In bulk materials, CDW phenomena are rather rare because of the unlikelihood of Fermi surface nesting. Uranium is the only element in the periodic table to exhibit a phase transition to CDW states at ambient pressure. The three phase transitions can be seen as a drastic change in the derivative of the electric resistivity vs. temperature. Interestingly, these phase changes do not lead to any changes to the orthorhombic structure, but primarily the changes are observed on the thermal expansion coefficient, the heat capacity and the electronic charge density. Studies on different materials have indicated that CDWs compete with superconductivity for conduction electrons, and this competition is hypothesized to be the reason for the absence of intrinsic superconductivity in -U. Attempts were made to understand the intrinsic superconductivity in -U in the absence of such phase transitions by increasing the external pressure, which was expected to suppress the phase transition. The increased critical temperature in single crystal -U with the application of pressure was precisely documented in, confirming intrinsic superconductivity in -U in the absence of CDW-caused phase transitions. However, some recent studies do strongly favor the presence of superconductivity in -U at ambient pressure. The average electrical resistivity of -U at room temperature is around 0.2 m which is similar to iron. Few researchers have measured the increased electrical resistivity of -U with respect to temperature. The electrical resistivity of -U is also anisotropic, with the direction having the highest resistivity and the lowest resistivity at elevated temperatures, although at low temperatures (<400 K), exhibits slightly higher resistivity compared to. Mechanical Properties The anisotropic nature of the mechanical properties in -U was the major motivation behind several early experimental and recent modeling efforts. The elastic constant variation with temperature in single crystals was experimentally determined by Fischer and is shown in Figure 3. All the elastic constants exhibit increasing negative curvature with temperature above room temperature, such that significant softening occurs as the beta phase transition temperature is approached. However, there is a second order transition in C11 at the low temperature CDW-induced phase change around 43 K. The moduli decrease monotonically with temperature above 273 K, reflecting the changes in the single-crystal constants. In addition, the polycrystalline mechanical properties were computed using single crystal elastic constants in by applying the Voigt-Reuss-Hill approximation. The Young's modulus in the direction (normal to the corrugated planes) is roughly 30% less than in or, indicating the relatively weaker bonding between corrugated planes versus within the planes. The Poisson's ratio of -U was also observed to be anisotropic and is not a strong function of temperature below 800 K. The mechanical properties of polycrystalline -U vary substantially with temperature, as shown in Figure 4. The tightly bonded corrugated basal planes in -U result in a very limited number of slip systems, such that room-temperature slip occurs only on these corrugated basal planes parallel to the direction of the corrugations ( system). This singular slip system is observed to support macroscopic ductility at room temperature only with the activation of twinning. Above approximately 525 K, additional slip systems such as and slip systems become active, and the material becomes softer and much more ductile. At temperatures below 273 K, slip becomes more difficult, even on the preferred system. A transition from ductile to brittle behavior accompanies this decrease in slip and increase in twinning. The mechanical properties of polycrystalline -U vary substantially with temperature, as shown in Figure 4. The tightly bonded corrugated basal planes in -U result in a very limited number of slip systems, such that room-temperature slip occurs only on these corrugated basal planes parallel to the direction of the corrugations ( system). This singular slip system is observed to support macroscopic ductility at room temperature only with the activation of twinning. Above approximately 525 K, additional slip systems such as and slip systems become active, and the material becomes softer and much more ductile. At temperatures below 273 K, slip becomes more difficult, even on the preferred system. A transition from ductile to brittle behavior accompanies this decrease in slip and increase in twinning. The mechanical properties of polycrystalline -U vary substantially with temperature, as shown in Figure 4. The tightly bonded corrugated basal planes in -U result in a very limited number of slip systems, such that room-temperature slip occurs only on these corrugated basal planes parallel to the direction of the corrugations ( system). This singular slip system is observed to support macroscopic ductility at room temperature only with the activation of twinning. Above approximately 525 K, additional slip systems such as and slip systems become active, and the material becomes softer and much more ductile. At temperatures below 273 K, slip becomes more difficult, even on the preferred system. A transition from ductile to brittle behavior accompanies this decrease in slip and increase in twinning. Thermal Properties Electronic charge carriers play a dominant role in the thermal properties of -U, such as thermal conductivity. The literature suggests that uranium follows the Wiedmann-Franz law for the proportionality between electronic conductivity and thermal conductivity, which states that the ratio of the electronic contribution of the thermal conductivity to the electrical conductivity of a metal is proportional only to the temperature. The first-principles calculations of Hin show that temperature-dependent thermal conductivity of both polycrystalline and single crystal -U follow similar trends as the electrical resistivity. Both the thermal conductivity and the electrical resistivity of single crystal -U exhibit anisotropic behavior. The average thermal conductivity of -U at room temperature is about 25 W/mK, which is approximately an order of magnitude lower than copper. Single crystal -U exhibits anisotropic thermal conductivity, and the conductivity along the direction (normal to the corrugated plane) is about 25% lower than along the direction. A comparison of DFT and experimental results on thermal conductivity are plotted in Figure 5f. A near linear increase in thermal conductivity from 300-900 K is observed, primarily because of the dominant electronic contribution to the thermal conductivity. At low temperatures, the influence of defects (e.g., point defects, impurities, and dislocations) can significantly impact the thermal conductivity of -U, and hence the observed discrepancies between experimental results at low temperature may be the result of differences in the sample chemistry and crystal defects. The thermal expansion coefficient values of -U are anisotropic, with different, positive coefficients along the and axes, but the values along the axis are negative at all temperatures. It should be noted that although many non-cubic crystal metals exhibit anisotropic thermal expansion, -U is extremely unusual in having a negative thermal expansion coefficient. The first X-ray diffraction data for the anisotropy of the thermal expansion above room temperature were reported by Bridge et al. in 1956 for polycrystalline -U. However, more reliable data from dilatometer measurements were provided by Lloyd for single crystals with different orientations (shown in Figure 5a). In addition, the thermal expansion coefficient of a polycrystalline -U is a strong function of pressure, but only varies significantly with temperature at low temperatures (<150 K) (Figure 5b,c). Some discrepancies in the measured thermal expansion may be due to the different impurity levels of the samples. Thermal Properties Electronic charge carriers play a dominant role in the thermal properties of -U, such as thermal conductivity. The literature suggests that uranium follows the Wiedmann-Franz law for the proportionality between electronic conductivity and thermal conductivity, which states that the ratio of the electronic contribution of the thermal conductivity to the electrical conductivity of a metal is proportional only to the temperature. The first-principles calculations of Hin show that temperature-dependent thermal conductivity of both polycrystalline and single crystal -U follow similar trends as the electrical resistivity. Both the thermal conductivity and the electrical resistivity of single crystal -U exhibit anisotropic behavior. The average thermal conductivity of -U at room temperature is about 25 W/mK, which is approximately an order of magnitude lower than copper. Single crystal -U exhibits anisotropic thermal conductivity, and the conductivity along the direction (normal to the corrugated plane) is about 25% lower than along the direction. A comparison of DFT and experimental results on thermal conductivity are plotted in Figure 5f. A near linear increase in thermal conductivity from 300-900 K is observed, primarily because of the dominant electronic contribution to the thermal conductivity. At low temperatures, the influence of defects (e.g., point defects, impurities, and dislocations) can significantly impact the thermal conductivity of -U, and hence the observed discrepancies between experimental results at low temperature may be the result of differences in the sample chemistry and crystal defects. The thermal expansion coefficient values of -U are anisotropic, with different, positive coefficients along the and axes, but the values along the axis are negative at all temperatures. It should be noted that although many non-cubic crystal metals exhibit anisotropic thermal expansion, -U is extremely unusual in having a negative thermal expansion coefficient. The first X-ray diffraction data for the anisotropy of the thermal expansion above room temperature were reported by Bridge et al. in 1956 for polycrystalline -U. However, more reliable data from dilatometer measurements were provided by Lloyd for single crystals with different orientations (shown in Figure 5a). In addition, the thermal expansion coefficient of a polycrystalline -U is a strong function of pressure, but only varies significantly with temperature at low temperatures (<150 K) (Figure 5b,c). Some discrepancies in the measured thermal expansion may be due to the different impurity levels of the samples. coefficient as a function of pressure (("Reprinted/adapted from Ref., 2022, Elsevier), (c) thermal expansion coefficient as a function of temperature: comparisons between the DFT studies and experiments [26, ("Reprinted/adapted from Ref., 2022, Elsevier), (d) specific heat capacity as a function of pressure ("Reprinted/adapted from Ref., 2022,Elsevier)), (e) specific heat capacity as a function of temperature ("Reprinted/adapted from Ref., 2022, Elsevier), (f) thermal conductivity (DFT studies adapted from ) compared with available experimental results (Adapted from Ref. ). Irradiation-Induced Growth and Swelling Irradiated polycrystalline -U was initially characterized macroscopically in the late 1940s and early 1950s, and samples were observed to grow in length and/or increase in volume (swell) at a rapid rate when irradiated. Understanding the dimensional instability arising from radiation is important for reactor component design. The early results considered the sample deformation to be a function of material fabrication, working temperature and thermal history. Some studies revealed that the swelling rate of -U could be reduced with small additions of low solubility alloying elements, especially silicon, aluminum or carbon. It was also observed that thermal cycling produces damage in unirradiated uranium roughly like the radiation effects in appearance. Pugh theorized that the mechanism behind irradiation-induced elongation was the same as thermal cycling. Though it is now known that there is apparently no relationship, at that stage it was hypothesized that thermal cycling could be a stand-in for irradiation. In polycrystalline -U, enormous irradiation-induced elongation was observed especially at high burnup. Note that the macroscopic growth in polycrystalline samples is not the same as the volume-conservative growth in single crystals, though the phenomena are related. We are careful to specify polycrystalline or single crystalline sample states in this discussion. The first detailed analysis of polycrystalline sample growth was presented by Kittel and Paine in 1958, on the irradiation growth of uranium that had been processed with different conditions. The irradiation was conducted at the Material Test Reactor (MTR) with irradiation temperatures up to 493 K and burnups up to 1.82 at% (no details were provided on sample purity). This study indicated that irradiation growth is a strong function of burnup. Around the same time, Paine and Kittel measured the volume-conservative irradiation growth rate with respect to burnup for single crystal -U specimens, though the irradiation temperature is not well known. This work determined that single crystal specimens grow in the direction, shrink in the direction, and remain the same in the direction, and that the overall volume of the crystal is conserved during irradiation growth. The discovery of irradiation growth in single crystal uranium immediately leads to a hypothesis for the multiple observed irradiation-induced deformation behaviors in polycrystalline material and the variation in behavior depending on the sample processing. The collective deformation behavior of multiple individual grains behaving anisotropically should be influenced by grain size and grain texture. Initial studies motivated by macroscopic engineering concerns were succeeded by more scientific endeavors to understand the physics of the irradiation growth behavior. During 1952-1962, three different mechanisms were postulated. The first mechanism, proposed by Pugh, combined the insights from Cahn on the observed twin and slip systems and the idea of local thermal expansion induced by fission spikes to propose that irradiation growth resulted from irreversible plastic deformation in the heated region surrounding a fission track. Interestingly, Pugh's theoretical predictions on anisotropic dimensional change were experimentally observed in most of the single crystal swelling studies. The second mechanism, proposed by Seigle and Opinsky, hypothesized that the lattice imperfections generated in -U during irradiation could anisostropically diffuse to and be eliminated at grain boundaries and free surfaces. In this case, the interstitial atoms would add to the lattice, producing an extension, and vacancies would subtract from the lattice, producing a contraction of the grain at its boundaries. If suf-ficient diffusion occurs and the flux remains unbalanced, macroscopic growth could be observed. Using simplified considerations of the interstitial positions in the -U lattice and the possible diffusion routes, they theorized that the maximum interstitial migration would be along the direction and the maximum vacancy migration would be along the direction, implying lattice growth along and lattice shrinkage along. This is qualitatively the same as the observed single crystal deformation, but the actual anisotropic diffusion behavior of interstitials and vacancies separately and their interacting behavior in a crystal remains to be determined to confirm or reject this mechanism. The third mechanism, proposed by Buckley, hypothesized that the irradiation-induced elongation and contraction of uranium crystals are the result of the anisotropic condensation of vacancy and interstitial defects in planar clusters within the volume associated with a U-235 fission event. Based on this hypothesis, planar clusters of interstitial atoms increase the dimension of a crystal in the direction by creating extra planes, and planar clusters of vacancies erode existing planes, causing contraction in the direction. Around the same time, Hudson et al. performed the first thin film transmission electron microscopic measurements and observed the formation of dislocation loops in -U after neutron irradiation. To date, the details of irradiation growth in -U remain unclear, including the exact mechanism or mechanisms of irradiation growth in single crystals, the single-crystal irradiation growth behavior at elevated temperatures, and the impact of interphase boundaries, grain boundaries and solid solution impurities. Loomis and Gerber carried out a detailed investigation and studied the irradiation-induced dimensional change in single crystals and lineage crystals (crystals with low-angle grain boundaries) over a wide range of temperature (shown in Figure 6). Note that in Figure 6, the growth coefficient is defined as the ratio of change in length to the initial length per fraction of U fission atoms. They also reported the markedly enhanced dimensional changes in the and directions below the irradiation temperature of 50 K. The enhanced dimensional change was hypothesized to be the result of increased point-defect mobility due to the CDW-state observed below 43 K. However, it remains unclear whether irradiation-induced crystal growth completely stops at high temperature. Hudson observed that the presumed dislocation loops were randomly dispersed at low temperature but showed a very marked coplanar distribution at high temperatures (>623 K). The advanced characterization of -zirconium, which also exhibits anisotropic growth, has suggested that defect clustering and interstitial and vacancy loop formation is most likely in that material. They proposed this is to be a sequential result of the mechanism of movement into rows by glide, as the ease of deformation by slip increases with temperature. Thus, it appears likely that the hypothesis of irradiation growth resulting from vacancy and interstitial clustering on preferential planes is correct. By 1958, it was not yet clear whether the irradiation-induced dimensional change in polycrystalline material was a result of irradiation growth due to induced lattice defects, or whether it is swelling induced due to fission gases. In fact, in the 1950s, irradiation growth and swelling were not understood as separate mechanisms, and the observed changes in -U were simply thought of as irradiation-induced dimensional changes. Later it was found that the swelling was responsible for majority of the dimensional change. At the time, there was a hypothesis that dimensional changes resulting from lattice defects would anneal out, as well. Kittel and Paine suggested that the irradiation-induced dimensional changes under 573 K were not due to swelling induced by fission gases, and further macroscale studies were carried out at different irradiation temperatures and burnups to elucidate the answer. A first meta-analysis on irradiation-induced swelling above 673 K was reported by Pugh in 1961. Pugh plotted the irradiation swelling versus burnup and observed significant spread in the data. It was presumed that this spread was due to differences in the sample geometry, irradiation conditions, and potentially other factors. Certain samples exhibited grain boundary cracking or catastrophic "breakaway" swelling. A breakthrough was made on the effect of irradiation temperature and -U swelling when the data from Pugh's analysis were re-analyzed by Granata and Saraceno. This analysis included the effect of irradiation temperature, the data exhibited a peak in swelling at intermediate temperatures. The focus then shifted towards capturing the dependence of swelling on irradiation temperature, which was previously only studied with respect to the burnup. This new understanding of the effect of temperature on swelling lead to additional careful microstructural investigations of irradiation effects on -U. Materials 2022, 15, x FOR PEER REVIEW 9 o and -U swelling when the data from Pugh's analysis were re-analyzed by Granata a Saraceno. This analysis included the effect of irradiation temperature, the data exh ited a peak in swelling at intermediate temperatures. The focus then shifted towards c turing the dependence of swelling on irradiation temperature, which was previously o studied with respect to the burnup. This new understanding of the effect of temperat on swelling lead to additional careful microstructural investigations of irradiation effe on -U. The most comprehensive and systematic study on the effect of irradiation and te perature on -U microstructure evolution was performed by Leggett et al.. In t work, irradiations of high-purity polycrystalline -U (<150 ppm of impurity by weig were performed over the range of 573-923 K and over a burnup range of 0.03-0.4 at% fast neutron flux up to 2 10 13 n/cm 2. Qualitatively different microstructures were served depending on the irradiation temperature, potentially indicating that several ferent mechanisms contribute to irradiation damage in the material. The observed mic scopic images at different irradiation temperatures are shown in Figure 7 compa against the unirradiated microstructure (Figure 7a). Below 623 K, a severely worked crostructure indicating plastic flow in the material was observed (Figure 7b), though major volume change was measured. Recent modeling indicates that stresses high enou to cause plastic deformation or void formation can result in polycrystalline -U with tremely low levels of burnup. In the range of 673-773 K (Figure 7c), a similar work microstructure was again observed, but jagged voids on the order of 1 micron also peared, apparently the result of grain boundary tearing. Recent modeling demonstra that this is likely due to the weaking of grain boundary at high temperatures. In range of 773-873 K (Figure 7d,e), they observed a few deformation twins and crysta graphically aligned pores on the order of few microns in size. However, the grain bou aries were essentially free of pores. Above 873 K, tiny spherical pores (again in the or of 1 micron) were observed throughout the material and more dominantly at the gr boundaries (Figure 7f). Figure 8 summarizes the complex irradiation swelling behavi as a function of temperature. The most comprehensive and systematic study on the effect of irradiation and temperature on -U microstructure evolution was performed by Leggett et al.. In this work, irradiations of high-purity polycrystalline -U (<150 ppm of impurity by weight) were performed over the range of 573-923 K and over a burnup range of 0.03-0.4 at% at a fast neutron flux up to 2 10 13 n/cm 2. Qualitatively different microstructures were observed depending on the irradiation temperature, potentially indicating that several different mechanisms contribute to irradiation damage in the material. The observed microscopic images at different irradiation temperatures are shown in Figure 7 compared against the unirradiated microstructure (Figure 7a). Below 623 K, a severely worked microstructure indicating plastic flow in the material was observed (Figure 7b), though no major volume change was measured. Recent modeling indicates that stresses high enough to cause plastic deformation or void formation can result in polycrystalline -U with extremely low levels of burnup. In the range of 673-773 K (Figure 7c), a similar worked microstructure was again observed, but jagged voids on the order of 1 micron also appeared, apparently the result of grain boundary tearing. Recent modeling demonstrates that this is likely due to the weaking of grain boundary at high temperatures. In the range of 773-873 K (Figure 7d,e), they observed a few deformation twins and crystallographically aligned pores on the order of few microns in size. However, the grain boundaries were essentially free of pores. Above 873 K, tiny spherical pores (again in the order of 1 micron) were observed throughout the material and more dominantly at the grain boundaries (Figure 7f). Figure 8 summarizes the complex irradiation swelling behaviors as a function of temperature. Around the same time, Angerman and Caskey reported microstructural changes in -U over the range of 403-823 K and at burnups of 0.05 and 0.77 at%. The reactor conditions and neutron flux were not provided. Overall, their observations were very similar to that of Leggett et al. though this study focused on a somewhat lower temperature range and the samples in this study were of lower purity (~1000 ppm impurities by weight). Like Leggett et al., Angerman and Casky also observed several twin systems in most grains and intersecting twins were observed in some areas. Slip within the grains also took place, as evidenced by the bending of the twins. At higher burnups, more distorted grain structures were observed, likely due to deformation by slip. Small fission gas bubbles were uniformly distributed in the samples irradiated below 673 K, similar to other observations. The authors observed the formation of irregular cavities in several samples irradiated above 723 K, which contributed significantly to the overall swelling. The metallographic analysis shown in Figure 9 reveals large cavities with a range of length (10-200 microns), especially at high irradiation temperature. The cavities were mostly found in grain boundaries or junctions with angular incisions. Angerman and Caskey reported pronounced variations in microstructure and cavitation (formation of mechanical cavities/voids) with burnup, as shown in Figure 9. At burnups below 0.1 at%, the grain structure of the uranium was easily recognizable regardless of the irradiation temperature, though some grain boundary cracking was observed at high irradiation temperatures. This grain boundary cracking is likely a result of thermal expansion stress at high temperature, hypothesized in. Thermal cycling experiments without irradiation found that polycrystalline samples deformed significantly and developed interior cracks with about 100 K temperature difference, and recent modeling has shown that thermal stresses arising from the negative thermal expansion coefficient is sufficient to cause cracking after a temperature change of about 100 K in irradiated -U. At burnups of 0.15 to 0.3 at%, the grain structure became twisted and distorted as the result of deformation by slip. Uniform and fine "marbleized" structures were observed at burnups of 0.6 to 0.8 at%, though data are only available only at temperatures below 623 K. to that of Leggett et al. though this study focused on a somewhat lower temperature range and the samples in this study were of lower purity (~1000 ppm impurities by weight). Like Leggett et al., Angerman and Casky also observed several twin systems in most grains and intersecting twins were observed in some areas. Slip within the grains also took place, as evidenced by the bending of the twins. At higher burnups, more distorted grain structures were observed, likely due to deformation by slip. Small fission gas bubbles were uniformly distributed in the samples irradiated below 673 K, similar to other observations. The authors observed the formation of irregular cavities in several samples irradiated above 723 K, which contributed significantly to the overall swelling. The metallographic analysis shown in Figure 9 reveals large cavities with a range of length (10-200 microns), especially at high irradiation temperature. The cavities were mostly found in grain boundaries or junctions with angular incisions. Angerman and Caskey reported pronounced variations in microstructure and cavitation (formation of mechanical cavities/voids) with burnup, as shown in Figure 9. At burnups below 0.1 at%, the grain structure of the uranium was easily recognizable regardless of the irradiation temperature, though some grain boundary cracking was observed at high irradiation temperatures. This grain boundary cracking is likely a result of thermal expansion stress at high temperature, hypothesized in. Thermal cycling experiments without irradiation found that polycrystalline samples deformed significantly and developed interior cracks with about 100 K temperature difference, and recent modeling has shown that thermal stresses arising from the negative thermal expansion coefficient is sufficient to cause cracking after a temperature change of about 100 K in irradiated -U. At burnups of 0.15 to 0.3 at%, the grain structure became twisted and distorted as the result of deformation by slip. Uniform and fine "marbleized" structures were observed at burnups of 0.6 to 0.8 at%, though data are only available only at temperatures below 623 K. The formation mechanisms of the multiple morphologies of irradiation-induced swelling within -U (jagged cavities along grain boundaries, small aligned or non-aligned pores, massive cavities) is still an area of active research. Historically, the research community was unsure whether small pores (round spaces observed in the matrix) (observed in ) were voids or fission gas bubbles; they were later confirmed to be voids. This uncertainty originated from the fact that no advanced electron microscopy was used in Figure 9. The formation mechanisms of the multiple morphologies of irradiation-induced swelling within -U (jagged cavities along grain boundaries, small aligned or non-aligned pores, massive cavities) is still an area of active research. Historically, the research community was unsure whether small pores (round spaces observed in the matrix) (observed in ) were voids or fission gas bubbles; they were later confirmed to be voids. This uncertainty originated from the fact that no advanced electron microscopy was used in these investigations, generally because they were still new tools or not even invented yet. Fission bubble growth was argued to be negligible at the burnups for which these pores were observed, though a few arguments were put forward to support the assertion that the observed pores were gas bubbles which then grew enormously, leading to high swelling above 723 K. Pugh suggested that the large "breakaway swelling" volumes were formed by the diffusion of gaseous fission products to alpha grain boundary cracks, ultimately driving plastic deformation of the matrix. Speight and Greenwood later proposed that dislocation sweeping could promote the breakaway condition by increasing the average bubble size through bubble coalescence. It was also demonstrated that enhanced swelling may arise in -U for >823 K if the intergranular stresses due to irradiation growth are high enough to move dislocations. Conversely, later studies show clear evidence that certain porosity is due to mechanical cavitation and void formation. Recent modeling work has indicated that mechanical tearing along grain boundaries can arise from thermal stresses and irradiation growth stresses, but the nucleation mechanism of intergranular voids remains unclear. Twin boundaries were implicated in the formation of crystallographically aligned voids, though later work hypothesized they are void superlattices. Fission gases and vacancy agglomeration are both considered as mechanisms for void formation in -U. To validate any of these hypotheses, the atomistic behavior of collision cascades and point defects in -U and their collective behavior at diffusional length and time scales at low burnups must be studied with advanced characterization and simulation techniques. In addition, although several experimental investigations indicate that impurities affect the irradiation growth and swelling rate, there remains no comprehensive analysis of the role of these impurities in controlling irradiation growth and swelling behavior. Irradiation Effect on Mechanical Properties Although several attempts have been made to understand the effect of irradiation on mechanical properties of -U, such as ductility, hardness and creep, the results of these studies have been contradictory. A common observation, however, is the increased hardness of -U, which was hypothesized to be a result of fission product impurity, irradiation-induced voids or dislocation loop formation. In 1958, Kittel and Paine tested polycrystalline natural uranium in the Materials Testing Reactor and found that the hardness increased with burnup, and that annealing did not restore the hardness. Similar observations were also made by Loomis et al. on single crystalline -U in 1964. However, a saturation in hardness was observed at a fluence of approximately 1 10 17 neutrons/cm 2. Both studies irradiated material at relatively low temperatures of up to 493 K and neither study provided information about the purity of the samples. Although radiation hardening is well known to occur due to the impediment of dislocation motion by defect loops, the reason for the observed yield strength saturation is unclear. The effect of irradiation on the creep and ductility of single crystal -U remains unclear. High-purity single crystals of -U were irradiated and then mechanically tested, and the single crystals retained considerable ductility (Figure 10a). Conversely, significantly lowered elongations were observed in polycrystalline -U by Vorob'ev and Konobeevskii et al. (Figure 10b). Konobeevskii et al. attributed such reduction to the formation of numerous internal cracks in polycrystalline metal as a result of irradiation. The initial study on creep testing in irradiated U was also reported by Konobeevsky et al., who irradiated the material and then creep-tested it out of pile. The creep rate was increased by a factor of 1.5 to 2 versus the un-irradiated material. Following this, Roberts and Cottrell conducted in situ creep testing in an irradiation environment, and observed a similar enhancement in creep during irradiation. Although the mechanical behavior of irradiated -U varies significantly within the literature, it appears that while single crystal material retains its ductility, the observed loss of strength and ductility in polycrystalline material is likely the result of the presence of grain boundaries. In fact, the possibility of superplasticity in irradiated polycrystalline -U has been discussed in. Though internal stress-driven plasticity (a phenomenon also found to be responsible for generating environmental superplasticity in other materials ) is proposed to contribute to the observations in, there is a need for more comprehensive mesoscale modeling of irradiation considering the effects of plasticity and grain-boundary interactions. Internal stress-driven plasticity under irradiation could arise from the anisotropic radiation growth of differently oriented grains, causing stresses high enough to initiate plastic flow. Further irradiation would continue to drive irradiation growth and stresses. Microcracks at grain boundaries could arise due to the presence of hard inclusions, insufficient slip and twinning processes to accommodate deformation across grain boundaries, or low grain boundary adhesion. However, detailed comparisons of mechanical behavior in-pile and out-of-pile from the existing literature may not be meaningful, given the very different samples, irradiation and testing conditions. Many historical tests did not characterize the details in the microstructure or composition of the materials. Many of the tests on irradiated material were performed before the development of irradiation damage models (Kinchin-Pease, Norgett-Robinson-Torrens) that calculate displacements per atom (dpa), and thus neutron fluxes, fluences, and spectra need to be reported to obtain an accurate understanding of the amount of irradiation damage. Furthermore, the irradiation temperatures were historically not well controlled or measured. Finally, given the difference in the purity level, grain size and fabrication methods, all of which could affect the irradiation behavior and mechanical response, more detailed testing is needed. Irradiation Effects on Thermal and Electrical Properties Neutron irradiation increases the electrical resistivity and thermal conductivity of -U, but the specific relationship is not well understood. A direct correlation was observed between the irradiation-increased resistivity and single crystal irradiation growth in all the available studies, albeit with limitations on the burnup range investigated. The effect of thermal neutron irradiation on the electrical resistivity of lineage single crystals (<600 ppm impurities) was studied by in the CP-5 reactor up to 550 K. For a constant Although the mechanical behavior of irradiated -U varies significantly within the literature, it appears that while single crystal material retains its ductility, the observed loss of strength and ductility in polycrystalline material is likely the result of the presence of grain boundaries. In fact, the possibility of superplasticity in irradiated polycrystalline -U has been discussed in. Though internal stress-driven plasticity (a phenomenon also found to be responsible for generating environmental superplasticity in other materials ) is proposed to contribute to the observations in, there is a need for more comprehensive mesoscale modeling of irradiation considering the effects of plasticity and grain-boundary interactions. Internal stress-driven plasticity under irradiation could arise from the anisotropic radiation growth of differently oriented grains, causing stresses high enough to initiate plastic flow. Further irradiation would continue to drive irradiation growth and stresses. Microcracks at grain boundaries could arise due to the presence of hard inclusions, insufficient slip and twinning processes to accommodate deformation across grain boundaries, or low grain boundary adhesion. However, detailed comparisons of mechanical behavior in-pile and out-of-pile from the existing literature may not be meaningful, given the very different samples, irradiation and testing conditions. Many historical tests did not characterize the details in the microstructure or composition of the materials. Many of the tests on irradiated material were performed before the development of irradiation damage models (Kinchin-Pease, Norgett-Robinson-Torrens) that calculate displacements per atom (dpa), and thus neutron fluxes, fluences, and spectra need to be reported to obtain an accurate understanding of the amount of irradiation damage. Furthermore, the irradiation temperatures were historically not well controlled or measured. Finally, given the difference in the purity level, grain size and fabrication methods, all of which could affect the irradiation behavior and mechanical response, more detailed testing is needed. Irradiation Effects on Thermal and Electrical Properties Neutron irradiation increases the electrical resistivity and thermal conductivity of -U, but the specific relationship is not well understood. A direct correlation was observed between the irradiation-increased resistivity and single crystal irradiation growth in all the available studies, albeit with limitations on the burnup range investigated. The effect of thermal neutron irradiation on the electrical resistivity of lineage single crystals (<600 ppm impurities) was studied by in the CP-5 reactor up to 550 K. For a constant irradiation temperature, electrical resistivity increased with irradiation dose and was also proportional to the increase in irradiation growth strain. However, the increase in resistivity with irradiation dose is reported to saturate, contrary to the dimensional changes induced by irradiation growth. Though the very few results about the effects of irradiation on the thermal properties of -U are non-conclusive given the uncontrolled testing performed in 1960s, the existing results and estimates do reveal that the thermal conductivity of irradiated -U is impacted. The overall dependence is governed by various phenomena including lattice resistance imposed by defects, microstructural damage on the grain boundaries, porosity development and the presence of fission gases. Brailsford and Major theoretically estimated the reduction in thermal conductivity as well as electrical resistivity due to irradiation in the form of an equation (Equation ), considering various key factors, where R is the % reduction in the thermal conductivity, D 0 and D 1 are the initial and final density (g/cm 3 ) (evaluated through any applicable swelling model) and b is burnup (at %). The burnup dependence implicitly accounts for changes due to lattice strains, lattice defects and fission gas bubbles and voids. Though the estimate considers several factors, several simplifying assumptions were made. Although the analysis considers the increased resistivity from irradiation effects, the effect of dislocation loops were totally neglected. However, the effect of dislocation loop dynamics is expected to play a vital role in the irradiation growth rate. Given the strong, suggestive results of the dependence of electrical resistivity on the irradiation growth rate and similar mechanisms responsible for thermal resistivity, more comprehensive models need to be developed. To date, there is no reliable experimental analysis to validate Equation. However, recent DFT studies by Peng et al. estimate the effect of U vacancies, U interstitials and Zr substitution on the thermal conductivity. Although Equation does not include temperature dependence, there is reasonable order-of-magnitude agreement between its predictions and the DFT studies. Among the three types of defects studied by DFT, U vacancies have the most substantial impact on thermal conductivity. Similar to the case of irradiation effects on dimensional changes and mechanical properties, further experimental investigations are required to understand the effect of irradiation on -U's thermal conductivity and electrical resistivity under carefully controlled conditions, including specification of the initial purity, the metallurgical condition of the specimen and the details of the irradiation (temperature, flux, fluence and neutron spectrum). Simultaneous measurements of the electrical resistivity and thermal conductivity during irradiation would be valuable to test the validity of the Wiedemann-Franz law for irradiated specimens, which forms the basis of Brailsford and Major's theoretical model. The effect of irradiation on the superconductivity of -U is a fundamental area of interest that could receive more attention. Existing results suggest that irradiation could affect the superconductivity of materials in two different ways: First, extended defects, such as column-shaped amorphous regions, act as strong, correlated pinning centers and usually increase the critical current density substantially. Second, point defects influence the microscopic parameters responsible for superconductivity, thus affecting the critical temperature and the critical fields. Analysis of the irradiation effects could provide a strong fundamental basis to characterize the intrinsic superconductivity of uranium as well as quantify the impact of irradiation-induced defects. Fundamental Research Needs The objective of this review is to consolidate and synthesize research efforts to date to understand the irradiation damage and effects in -uranium. This review surveys basic physical properties of -U and the effects of irradiation damage, including microstructural changes, dimensional changes, mechanical properties and physical properties. The anoma-lous and complex behaviors of -U are particularly highlighted, and further research in these topics will provide new insights into the irradiation damage mechanisms of -U. With advanced characterization and modeling techniques, several important research gaps can be investigated. Irradiation conditions (such as temperature and burnup), microstructure (such as grain structures and porosity), chemical composition (such as minor elements and impurities) and internal stress have profound effects on the irradiation response of -U. However, the understanding is generally qualitative in nature, rather than quantitative, and several mechanisms driving irradiation response still require explanation. Both experiments and modeling will be needed to answer these questions, and special care needs be taken to characterize the initial conditions of the material to determine quantitative behaviors. In single-crystal -U, the temperature-dependent irradiation growth behavior remains to be validated (reproduced with new experiments) and explained. Fundamentally, the mechanism of irradiation growth remains to be determined, and requires both advanced characterization (e.g., transmission electron microscopy and positron annihilation spectroscopy) and atomistic modeling and simulation (point defect migration and clustering behavior) to fully answer. Two other major questions are why the irradiation growth rate drastically increases at cryogenic temperatures and whether irradiation growth stops at temperatures near the phase boundary. The irradiation response of polycrystalline -U is driven by the interplay of single-crystal behaviors and the presence of grain boundaries. Recent modeling has indicated that grain boundary adhesion decreases at higher temperatures and that irradiation growth in polycrystalline material can cause stresses up to hundreds of megapascals, sufficient to drive plastic flow or cracking. However, the migration of point defects under stress fields and their interaction with sinks such as grain boundaries, voids and second phase precipitates remains to be studied. The effect of chemistry/impurities on the irradiation behavior is not well understood. Most of the available data are from experiments performed on samples with different impurity levels. As mentioned earlier, impurities have a profound impact on irradiation growth and swelling behavior. Certain impurities are known to reduce irradiation growth and swelling due to precipitate formation, solute-defect interactions or other such mechanisms. In addition, the impurities were also known to affect the nucleation of bubbles and cavities, leading to significant variations in mechanical behaviors. Highly pure -U specimens are recommended for benchmark irradiation studies. The role of internal stresses on sample auto-deformation or during mechanical testing needs to be further studied. Due to the significant thermal expansion anisotropy, bulk polycrystalline -U is likely always experiencing microstructural stresses, except after proper heat treatments. A temperature change of 50-100 K is sufficient to induce stresses on the order of hundreds of megapascals, large enough to drive plastic flow or cracking. Irradiation growth-induced stresses can also cause internal stresses on the order of hundreds of megapascals with very low levels of burnup. As a result, internal stress could be an important mechanism driving plasticity or superplasticity in -U.

Since the Aadhar mobile linking became compulsory, many people were facing problem in doing so, to solve the issues arising in the linking of the two and to make sure that no one, especially the senior citizens do not have any problem in doing the same, the government is making plans to modify the linking process and make it more convenient and easy for them to complete the process. Nirmal Singh 3C Company says that for easing the process, the government is planning to permit proxy authorization. After consulting from various telecom companies which include the use of online OTP (One Time Password) for mobile number that are being registered with the Unique Identification Authority of India, they are planning to give the customers alternatives like iris scanning for people who have indistinct fingerprints and even the facility of home visits for the senior citizens. According to the sources at Nirmal Singh 3C Company, UIDAI would be issuing guidelines and directions outlining the new options that they would be providing the mobile users for the ease in the linking of the Aadhaar. There were news stating the annoyance amongst the citizens of the country regarding the poor implementation of the policy and the inconvenience the people faced during the process. Nirmal Singh 3C Company says that the proxy authorization that the government is planning to come up with will see a person nominated by a senior citizen for verifying the Aadhar linkage to the mobiles. Currently the plan is only for the senior citizens but might be extended to everyone later on. The move will ease the problems that the people linking the Aadhaar were facing along with the troubles with the mobile service providers faced because of long queues in front of the outlet due to the issues like non- availability of machines. Many mobile phone consumers had complained about the multiple messages they had been receiving from the telecom operators to link their sim cards with Aadhaar or they would face disconnection, but this will also be looked upon by the government now, says Nirmal Singh 3C Company.

Phase-locking for maps of a torus: a computer assisted study. In the description of bifurcations in a family of maps of an n-torus it is natural to consider phase-locked regions in the parameter space that correspond approximately to the sets of parameter values for which the maps have invariant tori. The extreme case of phase-locking is resonance, where the torus map has a periodic orbit. We study a family of maps of an n-torus that only differ from a family of torus translations by a small nonlinear perturbation. The widths of the phase-locked regions for this family generally increase linearly with the perturbation amplitude. However, this growth varies to a higher power law for families of maps that are given by trigonometric polynomials (the so-called Mathieu-type maps). The exponent of the asymptotic power law can be found by simple arithmetic calculations that relate the spectrum of the trigonometric polynomial to the unperturbed translation. Perturbation theory and these calculations predict that typical resonance regions for the family of Mathieu-type maps are narrow elliptical annuli. All these results are illustrated in a number of numerical examples.

Optimal Bit and Power Direct Allocation Algorithm in OFDM Systems Based on the Regularity of Greedy Allocation Result This paper proposes a new optimal bit and power direct allocation algorithm. Based on the analysis of the regularity of the Greedy allocation result, we propose and prove a necessary and sufficient condition for the Greedy allocation result. According to this condition, a bit and power allocation algorithm aiming to allocate the total bits or total power at a time to all subchannels is proposed. The impact of SNR Gap on the allocation result and the technique to eliminate its impact are analyzed. Though iterations are needed to obtain the ideal allocation result, the computational complexity is lower than that of existing optimal algorithms in the literature, even lower than that of some suboptimal algorithms.

Effects of Vaginal Lubricants on In-Vitro Progressive Spermatozoa Motility. Vaginal lubricants are used to solve intercourse difficulties or as sexual enhancers, but recent reports raise questions about their safety in terms of fertility. In this study, twenty semen samples were tested against commercially available vaginal lubricants for progressive spermatozoa motility and vitality with varying exposure time intervals. Results showed that the vaginal lubricant which least affected progressive spermatozoa motility was the oil-based vaginal lubricant, which kept the mean percentage of progressive spermatozoa motility within the minimum normal range of 32%, following 60 minutes of exposure. The silicone-based vaginal lubricant produced similar results to the oil-based vaginal lubricant, however the progressive spermatozoa motility dropped below the minimum normal range within 60 minutes of exposure. The fertility lubricant did not produce mean progressive motilities that were within the normal minimum range at any of the three time intervals, producing poor results overall. The vaginal lubricant which produced the poorest results was the water-based, which immobilized all of the spermatozoa within 5 minutes of exposure and killed on average 95.23% within 60 minutes. Although further assessment is required, these results highlight potential fertility issues related to the formulation of commercially available vaginal lubricants.

COLUMBUS — Ohio State’s spring football game will be Saturday at Ohio Stadium. While spring games might sometimes seem like an antiquated relic from an earlier era, coaches like to see how young players react in front of crowds and fans like to watch spring games, so they’re not going away anytime soon. 1. Ryan Day without the training wheels. Yes, he coached three games last season when Urban Meyer was suspended. No, the spring game is not anything like a regular-season game. 2. The starting quarterback for Ohio State in 2019. Justin Fields, a transfer from Georgia, is expected to win the job. But redshirt freshman Matthew Baldwin, who spent last season observing practice from the sideline as he recovered from a high school knee injury, is also competing to be the starter, according to Day. 3. Six of the newest Buckeyes, the 2019 recruits who enrolled early and went through spring practice, led by 5-star wide receiver Garrett Wilson and 5-star defensive lineman Zach Harrison will be in uniform. 4. Former and current Buckeyes will be honored. Defensive lineman Dre’Mont Jones and offensive lineman Michael Jordan, both of whom have declared for the NFL draft, will be recognized for being named All-Americans during the game. Also, between the first and second quarter the 2018 team members will receive rings for winning the Big Ten championship and the Rose Bowl. 1. Tackling. The Buckeyes’ spring game last year resembled a touch football game and it won’t be any different this year. 2. A 3 ½ hour-game. The spring game will be four 10-minute quarters and there will be a running clock in the second half. coaches other than defensive line coach Larry Johnson, is gone. 4. Starters like offensive lineman Thayer Munford and safety Jordan Fuller, and several other players who are rehabbing injuries. Also, healthy starters who have nothing to prove, like defensive lineman Chase Young, will make only token appearances if they play at all. 5. A full house. Construction on C Deck at Ohio Stadium means that only 76,000 seats will be available for the spring game. So the attendance will not approach the announced crowd of 100,189 in 2016 or 99,391 in 2015.

Declining an opportunity to debate opponents on local TV, Rep. Larry Bucshon said Thursday he'll go to a Sullivan County Republican dinner instead. EVANSVILLE, Ind. — Declining an opportunity to debate opponents on local television, 8th District Rep. Larry Bucshon said Thursday he'll go to a Sullivan County Republican dinner instead. But that declaration was preceded — and accompanied — by confusion. Would Bucshon appear at the May 2 dinner? Or the debate? The veteran congressman hinted Monday that he might accept the debate invitation, which came his way weeks ago. But on Thursday, he said through a spokesman, it's final: He'll be at the dinner in Sullivan County. The confusion left one of Bucshon's opponents in a May 8 GOP primary election claiming the congressman is trying to duck him — and missing an important opportunity in the process. "He definitely is dodging me — and I don't blame him," said Richard Moss, a Jasper-based surgeon and Bucshon's opponent. The 7 p.m. debate on May 2, planned in partnership between the Courier & Press, WNIN FM and WNIN TV, is set to be locally televised at WNIN's new location in Downtown Evansville. Bucshon's GOP primary challengers, Moss and Rachel Covington, have already accepted. Not Bucshon. Not in the end. "Unfortunately, we've got a scheduling conflict that night," Bucshon spokesman Andrew Hansen said Thursday. "It's the Sullivan County (GOP) Lincoln Day dinner. So he's attending that. He's been doing all of those throughout the district, so he's going to have to go to that." But Bill Springer, chairman of the Sullivan County GOP, said about an hour later that Bucshon hadn't yet accepted the party's dinner invitation. "I'll just be honest with you, I just printed the tickets up today, and I just called his office yesterday — so, I haven't had a chance to call everybody, to be honest with you because I just got my confirmation on the (Republican U.S. Senate candidates)," Springer said. "I'm just now putting this all together right now." That triggered this response from Hansen: "(The Sullivan County GOP dinner) is on our calendar. (Bucshon) goes to all of them. I don't know why they would say that." About a half-hour later, Springer called the Courier & Press to revise his comments. Bucshon verbally accepted his invitation to attend the May 2 dinner two months ago, Springer said. He said he did not speak to Bucshon or his representatives after the first phone call. "(Bucshon) was originally going to be the speaker, but since I got (commitments from Republican U.S. Senate) candidates, I'm going to let those guys speak," Springer said. Bucshon will be there anyway, the Sullivan County GOP head said. But Bucshon wasn't so sure about that Monday when he was asked about his participation in the debate after touring the Evansville FedEx Ground Distribution Center just off U.S. 41. Without mentioning the event in Sullivan County, the 8th District congressman said he had a previous commitment on May 2 — but he said he might still participate in the debate. Moss said Bucshon didn't accept his invitations to debate when they ran against each other in 2016 — but then, he could not offer the prospect of a televised encounter. "That was a no-brainer for him. He was not going to try to legitimize me," Moss said. "But here, it's something that obviously, the biggest paper in the district is holding in the biggest town in the district — and on TV. I think he needs to go." Covington, who resides in Tokyo, Japan, could not be reached for comment Thursday. The Courier & Press sent Bucshon's campaign a formal invitation to participate in the debate against Moss and Covington on March 16. That was about a week after the congressman's campaign was verbally notified of the event. Moss and Covington accepted their invitations before the formal invitations were sent. Bucshon has consistently declined opportunities to debate Republican primary opponents. Asked about a month ago whether he would debate this year, he said no. But he held open the possibility of accepting an invitation from the Courier & Press. "I'm probably not going to have any debates prior to the primary," he said then. "I have over 90 percent name ID in my district, and my constituents pretty much know where I stand on the issues. "Honestly, I just don't feel that a debate prior to the primary adds anything to that discussion." Told that a Courier & Press debate invitation would be forthcoming, Bucshon softened his stance. "I don't want to be hypothetical. Let's see what happens," he said. Staff writer Zach Evans contributed to this report.

COLUMBUS – The Winnipeg Jets will be without shutdown centre Adam Lowry as they kick off an important four-game eastern road trip Sunday night in Columbus. Lowry, 25, has been suspended two games for a reckless high-stick of Nashville’s Filip Forsberg in Friday night’s 5-3 win at Bell MTS Place. He will miss the contest with the Blue Jackets and Tuesday’s tilt against league-leading Tampa Bay and is eligible to return Friday night in Carolina. Lowry had just absorbed a hit from Forsberg when he swung his stick towards the Predators winger, hitting him in the face. Forsberg was shaken up but not seriously injured. Lowry was given a two-minute penalty on the play, then called on the carpet Saturday by the NHL’s department of player safety announced Saturday. “While we understand Lowry’s contention that he did not intent to strike Forsberg in the face on this play, it is important to note that this is not a situation where a player simply loses control of his stick, or one where an off-balance player flails to try to steady himself. This is a reckless stick swing that strikes an opponent at a dangerous height. No matter where he intends the blow to land, Lowry must be in control and responsible for his stick,” the NHL said in the suspension video. "While we understand Lowry’s contention that he did not intent to strike Forsberg in the face on this play, it is important to note that this is not a situation where a player simply loses control of his stick, or one where an off-balance player flails to try to steady himself. This is a reckless stick swing that strikes an opponent at a dangerous height. No matter where he intends the blow to land, Lowry must be in control and responsible for his stick," the NHL said in the suspension video. Lowry has eight goals and nine assists in 62 games this season. He will forfeit $31,362 in salary as a result of the ban. Lowry was previously suspended for one game in 2014 for a hit from behind on Buffalo’s Patrick Kaleta. Lowry joins injured defencemen Dustin Byfuglien, Josh Morrissey and Joe Morrow as other pieces currently out of the lineup. As well, the status of No. 1 goalie Connor Hellebuyck for Sunday isn't clear after he missed Friday's game with the flu. The Jets had an off-day Saturday as they flew to Columbus. Lowry’s suspension will trigger other moves, including Bryan Little likely moving back to centre for the next two games. Little was on the wing with Lowry and Brandon Tanev in Friday night’s win. As well, one of Matt Hendricks or Par Lindholm will draw into the lineup to fill Lowry’s void. Both were obtained last Monday at the trade deadline but have been healthy scratches in the two games since. Winnipeg is currently 1-1-0 in a stretch of nine straight games against teams holding down a playoff spot as of Saturday. The Jets are coming off one of the more complete efforts of the season in Friday's win. Winnipeg fell behind 2-0 late in the second period, then rallied for victory. They lead the Central Division by one point over Nashville, although they have three games in hand on the Predators. "Really important because we liked the way that we were playing and it went against us a bit but we didn’t change the way we were playing. It was a fun game to be a part of. The bench was wired right and everybody was pulling and into the game. Really important. What would I be selling if it didn’t go our way? I would be telling you that I liked the way that we played and if we keep playing like that. We like that we’re moving forward and it’s great to get a win on that and have everybody (have) a piece of it," coach Paul Maurice said following the game. "It’s good that they got properly rewarded for it." The road ahead doesn't get any easier. Columbus should be a desperate team after they lost 4-0 on home ice Saturday afternoon to the Edmonton Oilers. They began the day tied with Pittsburgh for the final Eastern Conference wildcard playoff spot. Making the post-season is a must for a team which held on to pending UFA's Sergei Bobrovsky and Artemi Panarin at the trade deadline, then added three more UFA's in Matt Duchene, Ryan Dzingel and Adam McQuaid while trading away numerous draft picks and prospects in an attempt to go "all-in" this season.

Americans barely increased their spending in February following a weak January performance, strong evidence the severe winter will hold back the economy in the first quarter. Consumer spending rose 0.3 percent in February following an even weaker 0.2 percent rise in January, the Commerce Department reported. Spending on autos and other durable goods actually fell for February and much of the small gain reflected higher utility payments to pay heating costs. Analysts said consumer spending, which accounts for 70 percent of economic activity, should rebound in the spring as the weather improves. Standard & Poor's Ratings Services is lowering its credit rating on Target Corp. following weaker-than-expected fourth-quarter results that were dragged down by a massive data breach and a disappointing foray into Canada. The rating agency said it lowered its ratings one notch down to "A" from "A+." The rating is still four grades above speculative or junk status. S&P says the outlook is still "Stable," implying further changes are not imminent. The move comes a month after the nation's second-largest discounter reported its fourth-quarter profit fell 46 percent on a revenue decline of 5.3 percent. The retailer's shares closed Friday at $59.98, up 24 cents. BMW is celebrating its 20th anniversary of building cars in the U.S. by investing $1 billion in its plant in South Carolina to build two of its new X-series vehicles at the facility that ushered in a wave of foreign automakers building Southern plants. The German automaker will produce the X7, a larger SUV with three rows of seats. The company will also make the X4, a sportier version of the X3 coupe, and plans to build a plug-in hybrid version of its smaller X5 SUV. The $1 billion will be spent through 2016 at the plant in Greer, near Spartanburg. BMW said it will hire 800 additional workers, bringing total employment at the plant to 8,800 people. McDonald's is offering free coffee to its customers for a limited time as competition for the breakfast crowd intensifies. The world's biggest hamburger chain said participating U.S. locations will offer small cups of McCafe coffee at no charge during breakfast hours from Monday through April 13. The Oak Brook, Ill., company is hoping that the coffee giveaway will bring in new customers and get existing customers to come in more frequently.

People are almost two times more likely to leave a website with obvious mistakes, according to a study. NEW YORK: Yet another reminder that writing skills are essential for PR pros: Bad grammar and typos aren’t just eye-sores, they cost businesses prospective customers. Web visitors are almost two times more likely to leave a website after looking at a single page if they see grammar and spelling mistakes than sites with correct grammar and spelling, according to a study by Website Planet, a web services comparison site. The study looked at more than 5,000 site visits during the last week of February and the first week of March, a press release said. Subjects were presented with two nearly identical landing pages, one with typos and another with none. Bounce rates, or the percentage of visitors who leave a site after looking at a single page, for the typo-ridden landing page were 85% higher, the study found. Additionally, customers spent 8% less time on landing pages with "deliberate typos and grammatical errors," according to the study. This can affect a website’s position on Google. If more visitors leave after only looking at a single page, Google sees the site as less trustworthy and "lowers its position in the organic Search Engine Results Pages," the study said. Similarly, people are 70% less likely to click on Google Ads with typos, the study said. As a result, Google lowers their position. Google charges 72% more for ads with grammar errors and 20% more for ads with spelling mistakes. "What this study tells us is that PR professionals cannot ever let their guard down because consumers are unforgiving, ever critical, and use good spelling and grammar as a barometer for the ability of a company to deliver," said Shira Stieglitz, Website Planet's head of content and research. "It underlines the old adage that first impressions count, with consumers showing an unwillingness to give brands a second chance at making a good one." This story was updated with comment from Stieglitz on March 19.

BBC Radio 2 - Ken Bruce, The Michael Jackson top-selling 60 UK hits countdown continues! The Michael Jackson top-selling 60 UK hits countdown continues! The Michael Jackson 60 at 60 countdown continues. Plus Aloe Blacc, the Grammy and two-time BRIT-nominated singer-songwriter picks the Tracks of My Years. Ken continues the 'Michael Jackson 60 at 60’ countdown, celebrating the man and his music - ahead of what would have been his 60th birthday next week. Plus, Aloe Blacc, the Grammy and two-time BRIT-nominated singer-songwriter picks the Tracks of My Years. Plus, Aloe Blacc, the Grammy and two-time BRIT-nominated singer-songwriter from California picks the Tracks of My Years with songs from Bob Marley and Donny Hathaway. Aloe released his debut major-label album ‘Lift Your Spirit’ in 2014 to huge acclaim, where it hit the top five on the UK and US charts. It followed the incredible success of singles’ ‘I Need A Dollar’ and ‘Wake Me Up’, his collaboration with the late DJ/Producer Avicii which became the fastest-selling single that year and a chart-topper in 100 countries. His last album also featured the number one record ‘The Man’, which reached the top 10 in the US, Australia, New Zealand and South Africa. His new album includes the latest single 'Brooklyn In The Summer' which Aloe says is 'electric with all kinds of energy' as he looks to celebrate the love in his life with the new music. Jason Mraz picks the Tracks of My Years. Madonna - The Immaculate Collection. The Very Best Of The Jacksons. Club Classics From The 80's (Various).

Calculus at A-level and its understanding It is my contention that in the teaching of calculus at A level we have, all too often, been producing an understanding of calculusand, implicitly, a view of mathematicsthat is distorted and unrealistic. I am glad of the chance to air these criticisms in this contribution to the debate about the future of sixth form mathematics. (In this article, sixth form is synonymous with A level, whether studied at school or college.) Nevertheless, as I maintain later, the changes now being wrought in sixth form mathematics generally may well prove beneficial.

Factor XIIIa mediated attachment of S. aureus fibronectin-binding protein A (FnbA) to fibrin: identification of Gln103 as a major cross-linking site. In the present study we investigated the role of factor XIIIa reactive Gln and Lys sites of staphylococcal FnbA receptor in cross-linking reaction with alpha chains of fibrin. For this purpose we produced two recombinant FnbA mutants in which either a single Gln103 site (1Q FnbA) or all identified reactive Gln103, 105, 783, 830 and Lys157, 503, 620, 762 sites (4Q4K FnbA) were substituted with Ala residues. The results of FXIIIa-catalyzed incorporation of dansylcadaverine and dansylated peptide patterned on the NH2-terminal segment of fibronectin revealed that the reactivity of Gln substrate sites was drastically reduced in 1Q FnbA and 4Q4K FnbA mutants, while the reactivity of Lys substrate sites was only moderately decreased in 4Q4K FnbA. When it was tested in the FXIIIa-mediated fibrin cross-linking reaction, the 1Q FnbA mutant exhibited about 70-85% reduction in reactivity compared to that of the wild-type FnbA. These results demonstrate that FnbA participates in cross-linking to alpha chains of fibrin predominantly via its Gln103 reactive site. Several minor sites, including residues replaced in 4Q4K FnbA mutant, contributed to an additional 15-30% of the total fibrin cross-linking reactivity of FnbA. Comparison of amino acid sequences that follow the major reactive Gln site in FnbA and several known substrate proteins revealed that FXIIIa displays a preference for the glutamine residue in an xQAxBxPx sequence, where Q represents reactive glutamine, x is any amino acid residue, A is a polar residue, B is either valine or leucine, and P is proline.

Effects of Water Models on Binding Affinity: Evidence from All-Atom Simulation of Binding of Tamiflu to A/H5N1 Neuraminidase The influence of water models SPC, SPC/E, TIP3P, and TIP4P on ligand binding affinity is examined by calculating the binding free energy G bind of oseltamivir carboxylate (Tamiflu) to the wild type of glycoprotein neuraminidase from the pandemic A/H5N1 virus. G bind is estimated by the Molecular Mechanic-Poisson Boltzmann Surface Area method and all-atom simulations with different combinations of these aqueous models and four force fields AMBER99SB, CHARMM27, GROMOS96 43a1, and OPLS-AA/L. It is shown that there is no correlation between the binding free energy and the water density in the binding pocket in CHARMM. However, for three remaining force fields G bind decays with increase of water density. SPC/E provides the lowest binding free energy for any force field, while the water effect is the most pronounced in CHARMM. In agreement with the popular GROMACS recommendation, the binding score obtained by combinations of AMBER-TIP3P, OPLS-TIP4P, and GROMOS-SPC is the most relevant to the experiments. For wild-type neuraminidase we have found that SPC is more suitable for CHARMM than TIP3P recommended by GROMACS for studying ligand binding. However, our study for three of its mutants reveals that TIP3P is presumably the best choice for CHARMM. Introduction The determination of binding affinity is a central problem in computer-aided drug design which is a useful tool to search for potential leads for various diseases. The accuracy of estimation of the binding free energy bind of ligand to receptor depends on computational methods and modeling of receptor-ligand complex. The docking method is usually used for locating binding sites and virtual screening of potential drug candidates from large data bases. This approach suffers from low accuracy and its results usually have to be refined by more sophisticated methods based on the molecular dynamics (MD) simulation. In many cases MD methods can reproduce reliable results on binding free energy having acceptable correlation with experimental data. Among them Molecular Mechanic-Poisson Boltzmann Surface Area (MM-PBSA), thermodynamic integration (TI), linear interaction energy (LIE), linear response approximation (LRA), free energy perturbation (FEP), and steered MD methods are widely used. Each method should be considered carefully to compromise between CPU time efficiency and accuracy level. In modeling of biosystems in aqueous environment, it is important to develop appropriate force fields and water models. Force fields, which are given in the form of empirical potential energy functions, have been developed by different groups. Today OPLS, CHARMM, AMBER, and GROMOS are the most popular force fields for all-atom simulation of biomolecules. To describe aqueous environment, one can use various models such as SPC, SPC/E, TIP3P, and TIP4P. The adjustment of parameters of these models is based on their ability to reproduce the enthalpy of 2 The Scientific World Journal vaporization and the density of water. SPC/E is especially accurate for capturing experimental properties of water such as the diffusion coefficient and dielectric constant. Because SPC, SPC/E, TIP3P, and TIP4P are relatively simple and able to provide reasonable results, they are often employed for simulation of peptides and proteins. Previous studies have revealed that different force fields provide different estimations for bind. Recently, the role of water molecules in the binding process has been considered. GROMACS manual (http://www.gromacs.org/Support/Online Manual) suggests that for allatom simulation of biomolecules water model TIP3P is suitable for AMBER and CHARMM, while TIP4P and SPC are more appropriate, respectively, for OPLS and GROMOS force fields. However, what water model is the best fit for a given force field in computation of bind remains largely unknown. To shed light on this problem, in this paper we study the impact of combinations of four main water models SPC, SPC/E, TIP3P, and TIP4P with AMBER99SB, CHARMM27, OPLS-AA, and GROMOS96 43a1 force fields on the binding affinity of Tamiflu to the wild type (WT) of A/H5N1 neuraminidase (NA). We have chosen the NA-Tamiflu complex because A/H5N1 virus causes great damage to live poultry markets, especially being recognized as human transmitted virus. More importantly, the binding free energy of Tamiflu to NA has been experimentally determined and this gives us the opportunity to compare theoretical estimates with the experimental ones. Using MM-PBSA method we have shown that combinations AMBER-TIP3P, OPLS-TIP4P, and GROMOS-SPC are the best choice for estimation of bind of Tamiflu. This result is in agreement with the GROMACS recommendation, which is followed from force field development. Contrary to the GROMACS suggestion, it is shown that SPC is more suitable for CHARMM than TIP3P but this conclusion is valid for the wild type of NA. Our study of three mutants H274Y, N294S, and Y252H reveals that TIP3P is presumably the best choice for CHARMM as suggested by GROMACS. It is found that bind obtained by the OPLS force field is much less sensitive to water models compared to other force fields. The difference in water models seems to have the drastic effect in CHARMM modulating both the receptorligand interaction energy and hydrogen bond network in binding area. For all studied force fields, SPC/E is worse than other aqueous models overestimating bind. Crystal Structure of A/H5N1 NA and Parametrization of Tamiflu. The initial structures of A/H5N1 WT and mutants H274Y and N294S were obtained from Protein Data Bank with PDB ID 2HU4, 3CL0, and 3CL2, respectively. Y252H was derived by the corresponding mutation in WT structure using the mutagenesis module, integrated in PyMOL package. For Tamiflu we use the oseltamivir carboxylate type. Its charges and atom types, used for MD simulation, are described in detail in our previous work. Namely, for united-atom GROMOS96 43a1 force field, charges and atom types of oseltamivir were fully parametrized by Dundee PRODRG2.5 Server (Beta). For the remaining all-atom force fields, atomic partial charges for Tamiflu were derived by ESP charge. To obtain optimal geometry for electrostatic potential calculations, its structure is first optimized with the help of Gaussian98 using the B3LYP/6-31G * level of theory. Fitting charges to the electrostatic potential was subsequently done by the RESP method. Atom types for Tamiflu were derived from different modules to get along with each force field. ACPYPE and MKTOP were adjusted to provide suitable atom types in OPLS-AA/L, while for AMBER99SB and CHARMM27, atom types were named by ACPYPE and SwissParam, respectively. Water Models. Water, known as an indispensable solvent in almost chemical and biological reactions, has been built in different ways to obtain reasonable models for computational study. A water molecule is characterized by its geometrical parameters such as bond lengths and angles which could be kept rigid or flexible during simulation. Each model is parametrized with atomic partial charges of oxygen and hydrogen and assigned with dispersion and repulsion forces approximated by Lennard-Jones potential. Water models are categorized by the number of points used to shape them, by rigid or flexible structures, and by integration or not with polarization effects. There are 46 distinct models that have from 3 to 6 sites. However, only 3and 4-site models ( Figure 1) are often used in simulations of biological systems. For 3-site model like SPC and TIP3P, a water molecule is constructed by one oxygen atom and two hydrogen atoms. Each atom is assigned with atomic partial charge, but only oxygen is allowed to have the Lennard-Jones interaction with other atoms. The van der Waals (vdW) interaction among hydrogen atoms was not parametrized yet. Three-site models are known as rigid and have the experimental geometry of water, except SPC which has the ideal tetrahedral angle of water as 109.47, but not 104.5 ∘. SPC/E, an updated version of SPC model, adds an average polarization correction to the potential energy function, resulting in the better density and diffusion constant than SPC model. The four-site model TIP4P is a rigid planar four-site interaction potential for water (Figure 1), having a similar geometry to the Bernal and Fowler model. Here the negative charge is shifted from the oxygen atom to a point 0.15 along the bisector between hydrogen atoms. In this paper, we just limit our study to only four frequently used water models SPC, SPC/E, TIP3P, and TIP4P ( Figure 1). Their geometrical and physical characteristics are described in Table 1. Here H, O, and L are the partial charges of hydrogen, oxygen, and lone pair, respectively. and are the H-O-H and lone pair-O-H angles, while and are the well depth and vdW radius, respectively. Molecular Dynamic Simulations. Complex of NA-Tamiflu is placed in a triclinic box of around 12000 water The Scientific World Journal Figure 1: The 3-site (a) and 4-site (b) water models. The labels are explained in Table 1. Table 1: Physical properties of water models. All data is recorded at 25 ∘ and 1 atm. H, O, and L are the partial charges of hydrogen, oxygen, and lone pair, respectively. and are the H-O-H and lone pair-O-H angles, respectively. and are the well depth and vdW radius, respectively. molecules with 1 nm distance between the solute and box (a typical snapshot is shown in Figure S1 of supplementary material available online at http://dx.doi.org/10.1155/2014/536084). The receptor and ligand have 3832 and 5749 atoms in the united atom and all-atom models, respectively. Periodic boundary condition is imposed in three directions. We use 1.4 nm and 1.0 nm cut-off for vdW and electrostatic interactions, respectively. Long range electrostatic interaction was computed by the particle-mesh Ewald summation method. Equations of motion were integrated using a leap-frog algorithm with a time step 1 fs. The nonbonded interaction pairlist was updated every 10 fs with the cut-off of 1 nm. All systems were neutralized by adding counter-ions and then minimized to remove the local strain in protein upon addition of all hydrogen atoms and to remove bad vdW contacts with water. By using the conjugate gradient method for every 50 steps of steepest descent, minimization is converged when maximum force becomes smaller than 0.01 kJ/mol/nm. Then, all bonds of protein were restrained, leaving remaining parts to relax for 100 ps to obtain evenly distributed system. The temperature was gradually heated to 300 K during 100 ps with 5 kcal/mol harmonic restraints in all systems. The equilibration was next performed, coupled with temperature and pressure. Constant temperature 300 K was enforced using Berendsen algorithm under 50 ps NVT simulation with a damping coefficient of 0.1 ps. Parrinello-Rahman pressure coupling was used in 100 ps NPT run for 1 atm with the damping coefficient of 0.5 ps. Final NPT simulations of 20 ns were carried out for all force fields. Each force field is combined with four different water models SPC, SPC/E, TIP3P, and TIP4P, except GROMOS, which uses only SPC and SPC/E models. In total, we have 14 different models for the NA-Tamiflu complex. All simulations have been carried out in the GROMACS suit with Gromacs-4.5 package. Binding Free Energy Calculation by MM-PBSA. The details of MM-PBSA method are given in our previous work. Overall, in this method the binding free energy of ligand to receptor is defined as follows: where elec and vdw are contributions from electrostatic and vdW interactions, respectively. sur and PB are nonpolar and polar solvation energies. The entropic contribution is estimated using the normal mode approximation. From 20 ns MD simulation output only snapshots collected in equilibrium are used to compute the binding free energy given by. Measures Used in Data Analysis. The C root-meansquare deviation (RMSD) is employed to measure the deviation of receptor structures from the initial configuration. The Scientific World Journal Table 2: List of 50 residues surrounding the binding site. Ile117 Arg118 The hydrogen bond (HB) is assumed to be formed if the distance between proton donor (D) and proton acceptor (A) is less than 0.35 nm and the angle H-D-A is also less than 30 ∘. Definition of Binding Site. The binding pocket is defined as a space surrounded by 50 amino acids as shown in Figure S2. Our definition is compatible with that of Cheng et al.. The list of these amino acids is given in Table 2. Volume of the binding pocket is approximately estimated as volume of smallest convex hull which contains all of the fifty C atoms (Figure 2). The number of water molecules inside the convex hull is considered as the number of water molecules in the binding site. The binding pocket volume and number of water molecules inside it are calculated by Matlab software. The water density in binding site is a ratio of the number of water molecules to its volume. Results and Discussion In this section we present results obtained for WT NA if not otherwise stated. Equilibration Time Scales Depend on Force Field and Water Model. The time dependence of C -RMSD of NA in complex with Tamiflu is shown in Figure 3 for different sets of force fields and water models. The equilibration of system is reached when RMSD gets saturation. In AMBER the equilibration time eq ≈ 15 ns for SPC, while ≈10 ns is needed to equilibrate the system in other water models ( Figure 3). SPC gives the largest RMSD in equilibrium state. In OPLS RMSD reaches saturation quite fast, after about 3 ns for TIP3P and 5 ns for remaining models (Figure 3). TIP3P provides a bit larger departure from the initial structure compared to remaining models. For CHARMM eq ≈ 8 ns for all sets (Figure 3). The effect of water on stability of the NA-Tamiflu complex is at most pronounced in CHARMM, where TIP4P affects the stability to a greater extent than other models. In GROMOS one has different time scales for equilibration in SPC ( eq ≈ 7 ns) and SPC/E ( eq ≈ 10 ns), but in equilibrium the average values of RMSD are almost the same for both water models. Due to united-atom nature GROMOS is the most unstable having average value of RMSD ≈ 0.2 nm against 0.13 nm of other force fields (Figure 3). Effect of Water Model on the Receptor-Ligand Interaction Energy. The interaction energy int between ligand and receptor is shown in Figure 4. The most pronounced dependence on water is observed for CHARMM as int fluctuates at very different levels. Particularly, TIP3P and TIP4P models make the interaction energy highly unstable during the first 7 ns, while it remains almost stable during the whole MD run for SPC and SPC/E ( Figure 4). In equilibrium TIP3P and TIP4P give lower interaction energy than SPC and SPC/E. Average interaction energy int ≈ −259.2, −279.7, −228.5, and −216.9 kcal/mol for SPC, SPC/E, TIP3P, and TP4P, respectively. In GROMOS int, obtained by using SPC, is higher than SPC/E. The effect of water modeling is also visible for AMBER, where SPC/E provides the lowest interaction energy in equilibrium. int is almost the same in TIP3P and TIP4P ( Figure 4). As in the RMSD case (Figure 3), the results, obtained by the OPLS force field, are not affected much by water models (Figure 4). The combination of GROMOS with SPC and SPC/E gives the highest receptor-ligand interaction energy among four force fields, while the lowest int is obtained by CHARMM-SPC and CHARMM-SPC/E. Electrostatic Interaction Dominates over vdW Interaction in All Models. The separate contributions of these two interactions are shown in Figures S3 and S4. Clearly, the electrostatic interaction is far superior than vdW in binding affinity of Tamiflu to NA. This observation was reported previously for a few number of models, but the role of water has not been explored yet. Figure 3: C -RMSD of wild-type NA when interacting with Tamiflu during 20 ns simulations with different combination of force fields and water models. For AMBER equilibration time eq ≈ 15 ns for SPC (black arrow) while eq ≈ 10 ns for remaining water models (magenta arrow). In OPLS eq ≈ 3 ns for TIP3P (green arrow) and 5 ns for other models (magenta arrow). In the CHARMM case all systems reach equilibrium after about 8 ns. For GROMOS eq ≈ 7 ns (black arrow) and 10 ns (red arrow) for SPC and SPC/E, respectively. As follows from Tables 3-6, the contribution of vdW interaction to the binding free energy is not sensitive to water models for all force fields except CHARMM where SPC makes markedly higher contribution compared to other models. The effect of environment on the electrostatic interaction is weak in OPLS (Table 4) leaving elec almost equal in 4 water models. For AMBER (Table 3) SPC/E gives the lowest estimation for elec, while the drastic water effect is observed in CHARMM (Table 5) and GROMOS (Table 6). In the latter case two models yield the difference in elec of about 30 kcal/mol, but SPC/E and TIP4P in CHARMM provide even larger difference of ≈61.5 kcal/mol. Binding Free Energy Depends on Water Models. Apolar solvation energy sur and entropy contributions are not sensitive to force fields and water models (Tables 3-6). sur is about 4.5 kcal/mol, while − is 13-15 kcal/mol for all models. The dependence of bind on water mainly comes from competition between the electrostatic energy and polar solvation energy PB. If they compensate each other as in the case of AMBER and GROMOS, then the absolute value of bind is small (Tables 3 and 6). For OPLS PB is far below the absolute value of elec leading to large bind. This result suggests that the charge parametrization of OPLS is not suitable for studying binding affinity of oseltamivir to 6 The Scientific World Journal NA and its mutations. Since overestimation of bind was obtained by the MM-PBSA method, it remains unclear whether other methods would change this conclusion. The similar noncompensation effect is observed in CHARMM-SPC/E set (Table 5) where bind is also far below the experimental result (≈ −40.85 kcal/mol). SPC/E generates the most negative values for both electrostatic and vdW interactions compared with other 3 models ( Figure 4). Therefore, this model provides the highest binding affinity in all studied force fields (Tables 3-6). This observation agrees with the previous study of Hu and Jiang that the Coulomb interaction between water and lysozyme is more negative in SPC/E than in SPC and TIP3P since SPC/E has weaker self-diffusion than others, but closer to the experiment. In GROMOS force fields SPC and SPC/E give bind ≈ −11.79 and −18.56 kcal/mol, respectively (Table 6). Clearly, SPC result is closer to the experiment. Averaging the binding free energy over water models, one has bind = −18.36 ± 4.16, −66.96 ± 2.26, −28.33 ± 8.62, and −15.18 ± 4.79 for AMBER, OPLS, CHARMM, and GROMOS, respectively. Thus, the strongest effect of water modeling is observed in CHARMM as the departure from the average value bind is about 8.6 kcal/mol, while the weakest influence is seen in OPLS with deviation of ≈2.3 kcal/mol. Collin's group have shown that the binding free energy of Oseltaminir to A/H5N1 NA bind = −13.12 kcal/mol. Clearly, in AMBER TIP3P is the best fit to the experiments giving bind = −13.91 kcal/mol (Table 3). Thus, in accord with the GROMACS recommendation, AMBER-TIP3P is the best choice for studying ligand binding affinity. The agreement with the GROMACS's suggestion has been also obtained for GROMOS-SPC and OPLS-TIP4P (Tables 6 and 4) having bind closer to the experiments than other sets. It should be noted that OPLS-TIP4P is marginally better than OPLS-TIP3P because their difference in bind is less than 1 kcal/mol. So OPLS-TIP3P may not be a bad choice for estimation of binding affinity. For CHARMM the closest to experiment result ( bind = −17.79 kcal/mol) falls into SPC model (Table 5). TIP3P is ranked second having bind = −23.62 kcal/mol which is far from the experimental estimate. Thus, based on the results obtained for WT NA, one may recommend to use CHARMM-SPC instead of CHARMM-TIP3P suggested by the GROMACS. Since this conclusion is obtained for one system, to ascertain that SPC is the best choice for CHARMM we have computed bind for three more systems including mutants Y252H, N294S, and H274Y which have been studied experimentally. We summarize the main results in Table 7 providing details of calculations for different water models in Supporting Information (SI) ( Figure S5 and Tables S1-S4). The experiments show the ranking for binding affinity as Y252H > WT > N294S > H274Y. This ranking is correctly captured by TIP3P and TIP4P but not SPC as well as SPC/E (Table 7). Comparing absolute values of bind with the experiments one can see that SPC and SPC/E are the best for WT and Y252H, while TIP3P is the best for both N294S and H274Y. Taken together, in accord with GROMACS's suggestion, TIP3P is most suitable for CHARMM. Hydrogen Bond Network at the Binding Site. From previous MM-PBSA results, the hydrogen bonding, which mostly contributes to the electrostatic energy, plays the key role in the interaction between Tamiflu and A/H5N1 NA. However, the role of water has not been explored yet. Figure S6 shows the time dependence of HBs obtained by different force fields and water models. The HB number not only levels significantly among force fields but also depends on aqueous environments. Amber Force Field. Typical HB networks of four sets with AMBER are shown in Figure 5, where one has 7, 7, 6, and 7 HBs for SPC, SPC/E, TIP3P, and TIP4P, respectively. For all water models oseltamivir has the strong hydrogen bonding with residues E119, D151, R292, and R371 (lower panel of Figure 5). Within SPC/E the H-bonding with R152 is weaker than other models which have the population exceeding 80%. The strong interaction with E277 is observed only for this water model. Thus, in terms of individual contributions of ligand atoms SPC/E differs from other models. However, in equilibrium the average numbers of HBs are almost equal in all aqueous environments having HB( ) ≈ 6.6, 6.7, 7.0, and 7.0 for SPC, SPC/E, TIP3P, and TIP4P, respectively ( Figure S6). OPLS Force Filed. For OPLS four aqueous models show nearly the same HB networks ( Figure S6) Figure 5: Typical snapshots for hydrogen bond network between Tamiflu's charged groups and residues of NA at the binding site obtained by AMBER99SB force field with SPC (a), SPC/E (b), TIP3P (c), and TIP4P (d). Oseltamivir is hydrobonded with -COO − and -NH 2 (R371, R292); -OH (Y347); −NH 3 + and -COO − (D151, E119); NHAc and −NH 2 (R152) of NA. All hydrogen atoms are implicit. The lower panel refers to the probability of formation of HBs between ligand and receptor. The results are averaged over the last 2 ns of simulation. Black, red, green, and blue refer to SPC, SPC/E, TIP3P, and TIP4P, respectively. HB( ) ≈ 6.2, 6.0, 6.0, and 7.2 for SPC, SPC/E, TIP3P, and TIP4P, resp.). This is not surprising because they also have little effect on the binding free energy as discussed in the previous section (Table 4). HB patterns are quite similar among various water models in AMBER and OPLS force fields ( Figure 5 and Figure S7) implying that the geometry of ligand and area around the binding pocket does not depend much on water models. A slight difference is in population at residues R152 and E277 for two force fields. CHARMM Force Filed. The situation becomes very different in the case of CHARMM where water has the strong effect on the HB network ( Figure S8). The average number of HB in equilibrium HB( ) varies between 5.5 for SPC and 4.4 for TIP4P ( Figure S6). Contrary to AMBER and OPLS, only R371 remains the key residue for 4 aqueous models having the population more than 75%. SPC gives also strong H-bonding with E119 and D151 (population > 50%), while D151, R152, and R292 are H-bonded with Tamiflu for the most simulation time with SPC/E ( Figure S8). The ligand forms HB with R118 and E277 if one uses TIP3P but not other models ( Figure S8). TIP4P shows the modest HB population with E119, R152, and Y347, while together with R371 residue R292 is conserved in this water model. The diversity of HB networks in CHARMM presumably causes strong variation of the binding free energy among 4 aqueous models (Table 5). GROMOS Force Filed. As follows from Figure S6, due to the united-atom approximation used for this force field the number of HBs is much lower (HB( ) ≈ 0.7 and 1.6 for SPC and SPC/E) than other force fields. Consequently, HB networks are very poor ( Figure S9). H-bonding in SPC/E is stronger than SPC leading to its higher binding affinity (Table 6). Residue R152 has the substantial population in this water model. For SPC H-bonding is weak for all residues from the binding pocket. Table S5. The largest volume is obtained for SPC/E, while TIP4P provides the smallest volume. The time fluctuation of the number of water molecules inside the pocket (Figure 7) indicates that water molecules keep going out and coming back (see Movie 1). The weak dependence on water models is observed for AMBER force field because in equilibrium the binding pocket contains 42-46 water molecules (Table S5). SPC/E widens the binding site volume to a greater extent compared to other models providing the largest number of water molecules. Figure S10 shows the time dependence of the water density in the binding space bs w for all cases. As expected, bs w (Table S5) is lower than the standard density of 1 kg/L of water surrounding protein. It is well known that water weakens Hbonding leading to lower binding affinity than in vacuum. If this is true, then SPC/E model, for example, would provide the lowest binding affinity having the highest bs w. However, this is not the case as this model provides the highest binding affinity (Table 4). In general, one has the strong correlation (correlation level = 0.9) between bs w and bind (Figure 8) that the higher water density is, the higher binding affinity is. Since this correlation is at odds with the role of water in weakening H-bonding, one expects that HBs alone do not govern ligand binding affinity. Effect of Hydration on Using parameters of water models (Table 1), one can show that bind is not correlated with either the dielectric constant or dipole moment. Thus, one can not work out a unique factor that controls the binding affinity of Tamiflu to NA in AMBER. This is also true for other force fields. OPLS Force Field. As in the AMBER case, binding site volume (Figure 6), number of water molecules (Figure 7), and water density ( Figure S10) do not show much variations among water models. In equilibrium the volume fluctuates around 5000 3 for all water models. SPC shows the highest water density, while the lowest value of bs w is given by TIP4P (Table S6). The latter model also has the smallest binding site. SPC/E and TIP3P have the same water density (Table S6) but different binding free energies (Table 4). For four water 10 The Scientific World Journal models there is a modest correlation ( = 0.67) between bs w and bind (Figure 8). Again, similar to the AMBER case, this correlation cannot explain the binding affinity through the influence of water on HB network. CHARMM Force Field. The situation becomes entirely different in the case of CHARMM where the binding pocket volume ( Figure 6) and number of water molecules inside it ( Figure 7) are substantially higher than other force fields. TIP4P, which has the lowest volume in AMBER and OPLS, widens the pocket to the largest extent in CHARMM (Tables S5-S7). The number of water molecules in this model is about twofold larger than SPC/E. The packing of TIP4P water inside the binding site is also much denser ( Figure S10 and Table S7) than other models having bs w ≈ 0.37 kg/L. Nevertheless, the corresponding binding free energy remains higher than SPC/E (Table 5). Overall, there is no correlation between bs w and bind (Figure 8) in CHARMM. GROMOS Force Field. In GROMOS the binding pocket volume (Figure 6), the number of water molecules (Figure 7), and water density ( Figure S10) are lower than other force fields (Table S8). This presumably comes from united-atom approximation. There is a pronounced difference in the binding free energies obtained by SPC and SPC/E due to different water densities. Thus, similar to AMBER and OPLS, bind decreases with bs w. The Scientific World Journal Conclusions Our previous study on binding affinity of Tamiflu to variants of influenza A/H5N1 neuraminidase has revealed that AMBER99SB is the best among popular force fields as it reproduces bind with highest correlation and closest range with the experiments. In this paper, water models are tested with four force fields to examine their effects on the Tamiflu binding affinity toward the WT of A/H5N1 NA. As a result, TIP3P rather than SPC, SPC/E, and TIP4P goes along with AMBER99SB better than any combination of water models and other force fields. Within one force field, in agreement with the GROMACS recommendation, combinations AMBER99SB-TIP3P, OPLS-TIP4P, CHARMM-TIP3P, and GROMOS-SPC are more suitable for simulation of ligand binding. Although this result has been obtained for NA-Tamiflu complex and its validity for other systems is a subject for further investigation, the choice of these combinations is recommended. One has to bear in mind that the above combinations may not work in some cases. For example, CHARMM-SPC is a better choice for WT NA than CHARMM-TIP3P. In the case of Y252H, SCP/E works better than other water models if one utilizes CHARMM (Table 7). We have demonstrated that within one force field the binding free energy greatly varies for different combination of force fields and water models. For WT NA SPC/E always provides the lowest binding free energy among all water models regardless of force fields. Contrary to the remaining 3 force fields, bind estimated by OPLS-AA/L force field does not vary much among water models (Table 5). Due to strong electrostatic interaction, their values are too low compared with other force fields (Tables 3-6), but other terms are sensitive to them. The HB network between Tamiflu and NA changes little upon water models in OPLS and AMBER force fields, while it does strongly in GROMOS. CHARMM is a medium case. The pronounced influence of aqueous models on water density inside binding pocket has been observed in CHARMM force field.

Small and Strong MATERIALS SCIENCE The intricate silica cell walls fabricated by the unicellular algae known as diatoms are highly porous and are produced with high fidelity. Diatoms have therefore been viewed as a possible platform for nanostructured materials synthesis. Hildebrand et al. have probed cell wall synthesis in the nanostructured form of Thalassiosira pseudonana, an organism whose genome has recently been sequenced. They studied a series of structural intermediates to unravel the chemical formation sequence and to ascertain when certain proteins come into play. At the earliest stages, they observed an outline of the valve with silica ribs radiating from the center. The rim structure then thickens, followed by a thickening of the rest of the valve structure. As the ribs form and fuse together, they give rise to a nanoporous structure with larger, more irregular pores than those formed earlier in the process. These observations confirm that the structure of T. pseudonana has been optimized to maximize strength with minimized material requirements, all the while allowing for the uptake and efflux of metabolites during this process. The authors hope in the long term to replicate and control many of these features through modification of the genome or through mixing of an appropriate array of polypeptides and polyamines to foster silica polymerization in vitro. MSL J. Mater. Res. 21, 2689.

Mechanisms of action of Ru(ii) polypyridyl complexes in living cells upon light irradiation. The unique photophysical properties of Ru(ii) polypyridyl complexes make them very attractive candidates as photosensitisers in Photodynamic Therapy (PDT). However, to date, there are not many studies exploring in detail the mechanism(s) of action of such compounds in living systems upon light irradiation. This feature article provides an overview of the most in-depth biological studies on such compounds.

The present invention relates to orthopedic implants and more specifically to consolidated ceramic reinforced polymers useful in orthopedic implant applications. For at least three decades, ultra high molecular weight polyethylene (UHMWPE) has dominated the joint replacement arena as the material of choice in joint prostheses, i.e. primarily hip and knee replacements. The principle limitation controlling the longevity of hip prostheses is the body""s reaction to UHMWPE debris generated during interaction with the articulating femoral ball. The UHMWPE debris has been linked to bone resorption (osteolysis) which leads to loosening and premature failure of the implant. Failure of total knee replacements is largely associated with high stress fatigue loads that result in pitting, crack propagation and delamination of UHMWPE in such applications. For example, normal activities such as walking and climbing stairs can impose forces between the tibia and femur that are approximately four to five times ones body weight. Recent studies have shown that out of 937 tibial bi-compartmental and uni-compartmental bearings, 50% had indications of delaminations and cracking. In fact, total joint implants rarely last over ten years today. The rapidly growing aging population and the increasing use of implants in younger people provides a strong mandate for longevity improvements in total joint arthroplasties. Thus, numerous approaches to improving the useful life of such devices have been and continue to be proposed. Among the proposed improvements are; advanced crosslinking methods, carbon fiber reinforcement, and the use of a variety of counterface materials including CoCr, Ti-6Al-4V, and ceramics. Quite recently, surface engineering of metal counterfaces, e.g. coating, implantation, oxidation and diffusion has also been evaluated for reducing the wear rate of the preferred UHMWPE materials. Currently, most standard polyethylene components are irradiated with 5 Mrad of gamma radiation in a nitrogen atmosphere and the heated to 155xc2x0 C. for 24 hours to remove free radicals before being machined into components. Considerable research has been and continues to be expended to reduce wear rates and creep in UHMWPE since none of the foregoing methods has supplied a satisfactory solution. A method that uses high pressure to produce a more highly crystallized polyethylene (PE) (80% compared to 55% for normal irradiated PE) produced a material that exhibited higher yield stress, modulus and resistance to creep and crack growth, but neither laboratory wear tests nor clinical trials have demonstrated improvement in wear resistance. Numerous efforts have attempted to improve the performance of UHMWPE by increasing the cross-linking to improve its wear resistance. These efforts have included the application of higher doses of gamma radiation, electron beam radiation, and heating in combination with irradiation and chemical peroxide treatments to increase the degree of cross-linking. The mechanical properties of enhanced cross-linked PEs show a decrease in yield stress, elastic modulus, tensile strength, creep resistance, toughness and elongation to failure accompanied by an improvement in wear resistance. Since strength and creep resistance are crucial to extending the life of prostheses it is essential that these properties not be negatively impacted as other properties such as wear resistance are enhanced. Alternative materials have also been used to approach the solution to the prosthesis wear problem. Several different polymeric materials have, for example, been evaluated over the years. Polytetrafluoroethylene, various polyesters and polyacetyls have been evaluated without finding a successful replacement for UHMWPE. Chopped carbon fiber reinforced PE while demonstrating promising results in bench scale screening and simulator tests failed in clinical trials. Chopped carbon fiber reinforced epoxy-based cups articulating against alumina heads have demonstrated five times lower wear rates than UHMWPE cups in early clinical trials. In summary, although a large number of solutions have been proposed to provide improved hip and knee replacement materials, UHMWPE remains the current, if not entirely satisfactory, material of choice in such applications and the provision of an improved material or combination of materials which would provide improved wear resistance and strength for such uses remains an elusive objective. It is therefore an object of the present invention to provide an improved material for use in orthopedic implants. It is another object of the present invention to provide such an improved material that exhibits enhanced wear resistance and strength over existing and currently used UHMWPE materials. It is yet a further object of the present invention to provide improved orthopedic implants that exhibit useful lives beyond those implants currently in use today.

Study on the Influence of AC Stray Current on Buried Long Distance Pipeline With the rapid development of China's national economy and long-distance pipeline, the problems of parallel and cross laying of pipelines, transmission lines and railways are becoming increasingly prominent. AC interference corrosion threatens the safety of tube body, anticorrosive coating and cathodic protection equipment. The widespread application of UHV transmission lines puts forward higher requirements for pipeline design and operation safety. In order to reduce the harm of high-voltage transmission lines to pipelines, the mechanism of the influence of ac stray current on pipelines was analyzed, an experimental platform of AC interference corrosion was built, the influence of ac stray current density on corrosion rate was studied by weight loss method of corrosion specimens, and the protection measures and evaluation conditions of stray current were proposed.

The Clinical Significance of HbA1c in Operable Chronic Thromboembolic Pulmonary Hypertension Background Glycosylated hemoglobin A1c (HbA1c) has been proposed as an independent predictor of long-term prognosis in pulmonary arterial hypertension. However, the clinical relevance of HbA1c in patients with operable chronic thromboembolic pulmonary hypertension (CTEPH) remains unknown. The aim of the present study was to investigate the clinical significance of HbA1c as a biomarker in CTEPH. Methods Prospectively, 102 patients underwent pulmonary endarterectomy (PEA) in our national referral center between March 2013 and March 2014, of which after exclusion 45 patients were analyzed. HbA1c- levels, hemodynamic and exercise parameters were analyzed prior and one-year post-PEA. Results 45 patients (BMI: 27.3 ± 6.0 kg/m2; age: 62.7 ± 12.3 years) with a mean pulmonary arterial pressure (mPAP) of 43.6 ± 9.4 mmHg, a pulmonary vascular resistance (PVR) of 712.1 ± 520.4 dyn*s/cm5, a cardiac index (CI) of 2.4 ± 0.5 l/min/m2 and a mean HbA1c-level of 39.8 ± 5.6 mmol/mol were included. One-year post-PEA pulmonary hemodynamic and functional status significantly improved in our cohort. Baseline HbA1c-levels were significantly associated with CI, right atrial pressure, peak oxygen uptake and the change of 6-minute walking distance using linear regression analysis. However, using logistic regression analysis baseline HbA1c-levels were not significantly associated with residual post-PEA PH. Conclusions This is the first prospective study to describe an association of HbA1c-levels with pulmonary hemodynamics and exercise capacity in operable CTEPH patients. Our preliminary results indicate that in these patients impaired glucose metabolism as assessed by HbA1c is of clinical significance. However, HbA1c failed as a predictor of the hemodynamic outcome one-year post-PEA. were analyze (Fig 1). Baseline and follow-up right heart catheter (RHC) were not mandatorily performed in-house, as the Kerckhoff-Clinic is a national referral center. Patients with symptomatic chronic thromboembolic disease and mean pulmonary artery pressure <25 mmHg at baseline were excluded (Fig 1). Patients with a known history of diabetes mellitus or use of anti-diabetic treatment were excluded. All included patients gave written, informed consent, and the study was approved by the by the ethics committee of the Faculty of Medicine at the University of Giessen (Approval No. 31/13). All CTEPH patients were diagnosed according to current guidelines at that time and a multidisciplinary board including pulmonary physicians, PEA surgeons and pulmonary radiologists assessed operability. At inclusion, all patients had received oral anticoagulants for at least 3 months. All patients underwent PEA according to the protocol of the Kerckhoff-Clinic and treatment with targeted PAH therapy was permitted without restrictions. Residual PH one year post PEA was defined by mPAP >25 mm Hg and PVR > 240 dyne s/cm 5 at rest while the CTEPH type was classified by the surgical specimen as described previously. HbA1c testing Venous blood samples were collected at the time of initial referral and were immediately analyzed in the laboratory of the Kerckhoff-Clinic. HbA1c-levels were measured by Bio-Rad VARIANT™ II TURBO (Bio-Rad Laboratories, Inc. Hercules, California, USA) with a reference range of 29-42 mmol/mol. In addition, fasting plasma glucose, N-terminal fragment of pro-brain natriuretic peptide (NT-proBNP) and serum creatinine were measured. Glomerular filtration rate was calculated using the Chronic Kidney Disease-Epidemiology Collaboration (CKD-EPI) equation. The investigator who determined HbA1c was unaware of the patient baseline parameters or clinical course and HbA1c-levels were not used to guide patient management or monitor the effects of treatment during the study period. Baseline parameters Hemodynamic, 6-minute walk test (6MWT), echocardiography and cardio-pulmonary exercise testing (CPET) data from all included patients were collected before and after PEA. Right heart catheter (RHC) at baseline and follow-up were performed according current recommendations at that time. However, as the Kerckhoff-Clinic is a national referral center, RHC were partly performed at referring centers. All patients performed a symptom-limited incremental CPET using a ramp protocol with an incremental rate of 5 to 15 W/min judged by the operator (Masterscreen1, Carefusion1). In all patients 6MWT was performed according to current guidelines and patients were instructed to walk at their own pace along a 30-m corridor while standard phrases were communicated. Pulmonary artery systolic pressure (PASP) and tricuspid annular plane systolic excursion (TAPSE) were assessed by right heart echocardiography as described previously. Statistical analysis Data are presented as mean ± standard deviation for normally distributed parameters. As appropriate the 2-tailed T-test, paired T-test, Mann-Whitney-U-test, Wilcoxon Signed Rank test or Pearson Chi-Square test was used to test for differences between groups. HbA1c was dichotomized at the cut-off indicative for the diagnosis of diabetes (47.5 mmol/mol) and of pre-diabetes (39 mmol/mol) as stated in the position statement of the American Diabetes Association (ADA). Linear regression analysis was used to assess associations of HbA1c with clinical parameters. All baseline parameters were analyzed in a univariate logistic regression model with residual PH as dependent variable. Then, backward stepwise multivariate logistic regression models were used to predict residual PH one-year post-PEA, considering all parameters of the univariate analysis with a p-value <0.15. Statistical analysis was performed using SPSS, version 21.0 (IBM, Armonk, NY). Results Baseline 45 CTEPH patients with a mean age of 62.7 ± 12.3 years and a mean body mass index of 27.3 ± 6.0 kg/m 2 were included. Patients presented mostly in WHO functional class III with an impaired exercise capacity. At mean they had severe precapillary pulmonary hypertension with a mPAP of 43.6 ± 9.4 mmHg, PAWP of 9.9 ± 5.3 mmHg, PVR of 712.1 ± 520.4 dyne s/cm 5 and a substantially reduced CI. Right heart echocardiography showed an elevated PASP of 75.4 ± 24.1 mmHg and a reduced TAPSE of 17.6 ± 5.0 mm. At baseline the HbA1c was 39.8 ± 5.6 mmol/mol and NT-proBNP-levels were significantly elevated while GFR and serum creatinine were within the normal range. At time of inclusion 15 patients received monotherapy with pulmonary vasoactive therapies, mostly with phosphodiesterase type-5 inhibitors (Table 1). One-year post pulmonary endarterectomy outcome One-year post-PEA functional and hemodynamic parameters significantly improved and most patients were in WHO functional class II (44.4%). HbA1c, Glucose and serum creatinine remained unchanged while NT-proBNP and Hb were significantly reduced (Table 1). Residual PH was found in 24 patients with significantly elevated mPAP, PVR and NT-proBNP in comparison with non PH. In addition, a higher percentage of patients with residual PH presented in WHO functional class III and were treated notably with soluble guanylate cyclase stimulators (Table 2). No differences in HbA1c, Glucose or serum creatinine were evident in comparison of patients with residual and non-residual PH. Univariate logistic regression analysis revealed that baseline HbA1c-levels were not associated with the presence of residual PH one-year post-PEA (p = 0.22). Additional multivariate analysis including all parameters with p-value <0.15 of the univariate analysis and HbA1c (albeit not significant in the univariate model) revealed that HbA1c was not related to the hemodynamic outcome one-year post-PEA with a HR of 0.91 (95% CI for HR 0.76-1.01, p = 0.31). The multivariate analysis identified baseline peak VO 2 as a significant predictor of residual PH with a HR of 0.78 (95% CI for HR 0.62-0.99, p = 0.04), while other parameters failed to serve as independent predictors of the functional outcome one-year post-PEA in the stepwise backward multivariable model (Table 4). Discussion The present study prospectively evaluated HbA1c as a biomarker in a selected cohort of operable CTEPH patients and investigated its association with the outcome one-year post-PEA. An association of HbA1c with baseline hemodynamic parameters and exercise capacity was found. However, baseline HbA1c levels did not predict the hemodynamic outcome one-year post-PEA. Our study included operable CTEPH patients with severely impaired pulmonary hemodynamics and low exercise capacity mostly in WHO functional class III. Hemodynamic parameters and exercise capacity of our cohort mirrored the typical findings of patients with operable CTEPH prior to PEA with a substantial impairment of right ventricular function as well as elevated PVR, mPAP and reduced CI. One-year post-PEA hemodynamic parameters, exercise capacity and Nt-proBNP improved significantly, albeit 25 patients (49%) of our cohort exhibited a residual PH. In line with previously published data a significant improvement of the functional outcome one-year post-PEA was observed. However, as described in our study, the individual outcome after PEA differed with 10% to 24% of patients remaining symptomatic and up to 35% were presenting with residual PH. The established Table 1. doi:10.1371/journal.pone.0152580.g003 diagnosis of diabetes mellitus type II was present in 6 patients, which were excluded from the analysis. Nevertheless, the low prevalence of established diagnosis of diabetes type II in our cohort is in accordance with international registry data reporting diabetes type II in approximately 5.3% of CTEPH patients. However, using the ADA criteria 5 additional patients who did not have the diagnosis of diabetes mellitus yet, presented with indicative high HbA1clevels > 47.5mmol/mol. Interestingly, our data indicated for the first time that HbA1c-levels prior to PEA correlated with baseline RAP, CI and peak VO 2 in operable CTEPH patients, although similar hemodynamic parameters (apart from RAP) and RV systolic function were found in comparison of low HbA1c-levels (< 47.5mmol/mol) with elevated HbA1c-levels. In addition, patients with HbA1c levels > 39 mmol/mol, presented with a further impaired CI and higher RAP in comparison. The significant association of HbA1c with baseline RAP in CTEPH underlines the presence of diastolic dysfunction due to stiffer right ventricles in a more profound glycemic environment, as described for PAH patients previously. In addition, impaired glucose metabolism is potentially influencing CI in CTEPH by aggravating impaired myocardial relaxation and increased myocardial stiffness as seen in diabetic cardiomyopathy or metabolic disease in PAH. Similar to our finding of an association of glucose intolerance in operable CTEPH with worse exercise capacity as assessed by peak VO 2, Pugh and coworkers demonstrated a trend to a correlation of baseline HbA1c-levels with functional capacity (assessed by 6MWD) in PAH. Moreover, our study showed a weak but significant association of HbA1c-levels with the change of 6MWD one-year post-PEA. However, up to now, no robust associations of the altered glucose metabolism and 6MWD have been reported in PH patients. Zamanian and coworkers described no significant difference of the 6MWD between PAH patients with and without IR. Further, our data suggest that HbA1c is more elevated in advanced disease, although differences in WHO functional class, mPAP and PVR were not statistically significant. Despite these significant associations to baseline parameters, HbA1c was not significantly correlated to outcome parameters post-PEA in univariate and multivariate analysis. One can speculate that the influence of the glycemic environment pre-PEA on secondary small-vessel arteriopathy and RV dysfunction plays only a minor role one-year post-PEA and/or is not reflected by HbA1c directly. In addition, our cohort presented with predominantly proximal disease as reflected by the absence of Jamieson-Type IV patients. Furthermore, the relatively high number of residual PH in our study potentially results from incomplete removal of obstructions and/or a varying degree of reverse right ventricular remodeling after surgery rather than from concomitant small vessel disease. Thus, the prognostic impact of impaired glucose metabolism as a solitary marker is comparably moderate. Other pre-operative predictors of post-operative outcome such as mPAP, serum creatinine level, the number of involved segments, PVR and gender are useful complementary parameters to the currently reported HbA1c-levels. A limitation of our study is the lack of systematic evaluation post-PEA to differentiate between non-removable material or secondary microvascular disease as major cause of residual PH. In addition, register analysis featured an enhanced influence of diabetes as a risk factor in IPAH patients rather than operable CTEPH. Therefore, in case of inoperable CTEPH with distal disease and pronounced secondary small-vessel arteriopathy the association of impaired glucose metabolism assessed by HbA1c with pulmonary hemodynamics and long-term outcome remains unknown and in need of further investigations. Pathophysiologically, the obstructive remodeling of pulmonary arteries in CTEPH involves inflammation, in situ thrombosis and endothelial dysfunction. Inflammatory mediators play a key role in the pathological mechanisms of CTEPH partly mediating the obstructive remodeling of pulmonary arteries and being associated with RV dysfunction. Yet, cytokines have not been linked to impaired glucose metabolism to date. However, in patients with PAH and idiopathic PAH, albeit not in CTEPH, various studies have extensively analyzed the clinical relevance of HbA1c and an association of impaired glucose metabolism with disease severity was found. Moreover, IR has been identified as an important risk factor for disease progression and is associated with a poorer survival in PAH. IR is linked to pro-inflammatory cytokines with significant contribution to the pathophysiology of PAH and is associated with the development of pulmonary vascular disease in mice. Furthermore, animal models have shown that impaired glucose metabolism and IR have a direct impact on the pulmonary vasculature and the pulmonary smooth muscle cell (SMC) leading to higher RV pressures, enhanced RV hypertrophy and peripheral pulmonary arterial muscularization. In addition, the glycemic environment induces endothelial dysfunction with an impaired relaxant response through dysregulations in the NO pathway in diabetic rats. We did not asses IR as our study was not designed to evaluate these parameter, therefore future trails addressing this issue are warranted. Limitations of the study are the moderate sample size, the lack of prospective inclusion of inoperable CTEPH patients and the overall low prevalence of impaired glucose metabolism in our cohort. As we are a national referral center, the one-year follow-up visit was not mandatorily performed in our center and accounts for the dropout rate of 29 patients. In addition, the rate of residual PH was higher than reported in the literature. The relatively high rate of residual PH is not fully understood. The rate is influenced by many factors, including the type of pre-surgical CTEPH (proximal vs. distal), the severity of pre-surgical PH (possible reflection of micro-vascular disease) and the different underlying confounders that lead to CTEPH (e.g. simple recurrent deep vein thrombosis and pulmonary embolism vs. splenectomy vs. infected pacemaker leads etc.). We also believe that the selection criteria for the current observation might result in an enrichment of patients who are prone to have a comparably high proportion of residual PH after surgery. Lastly, a selection bias might have played a role, in that patients with residual PH (i.e. symptomatic patients) are more likely to be followed up by means of RHC. Conclusions In conclusion, this is the first study to describe an association of HbA1c-levels with pulmonary hemodynamic parameters and exercise capacity in a selected cohort of operable CTEPH patients. Our preliminary results indicate that in these patients impaired glucose metabolism as assessed by HbA1c is of clinical significance. However, HbA1c did not predict hemodynamic outcome one-year post-PEA in our cohort. Supporting Information S1 Table. Correlations of HbA1c with baseline parameter using linear regression analysis. (DOCX) S2 Table. Correlations of HbA1c with hemodynamic and functional outcome in patients with non-PH one-year post-PEA using linear regression analysis. (DOCX)

Slime production as a virulence factor in Staphylococcus epidermidis isolated from bacterial keratitis. A total of 126 coagulase negative staphylococci (CONS) isolated from corneal scrapings of patients of bacterial keratitis and 50 isolates from healthy eyes (controls) were tested for slime production. Eighty eight (69.84%) of 126 isolates from patients and 11 (22%) of 50 isolates from controls were slime producing (P < 0.001). Of these 88 isolates, 42 were Staphylococcus epidermidis biotype II, 30 were S. epidermidis biotype I, 8 were S. epidermidis biotype III and the rest belonged to CONS other than S. epidermidis. Amongst the corneal ulcer isolates, multidrug resistance (resistance to 3 or more antibiotics) was observed in 82.9 per cent (73/88) slime producing organisms as against only 18.4 per cent (7/38) nonslime producing organisms (P < 0.001). Similarly, of the total 99 slime positive and 77 slime negative isolates, 79 (79.8%) and 22 (28.6%) respectively were multidrug resistant (P < 0.001). Although, slime production is known to be one of the major virulence factors of CONS in extraocular systemic staphylococcal infections, the present study detected slime in isolates from ocular infections. It was found that S. epidermidis I and II were the common biotypes associated with bacterial keratitis and, slime production and multidrug resistance were the two important virulence factors. These observations have clinical and therapeutic significance.

A U.K. subsidiary of Rupert Murdoch&apos;s News Corp apologized in court here and agreed to pay damages to a former British intelligence officer whose computer was hacked by a private investigator working for the now-defunct News of the World. A spokeswoman for News UK, a News Corp subsidiary, said its News Group Newspapers business apologized in court Friday to Ian Hurst and accepted "vicarious liability," after a private investigator for the News of the World got access to his emails in 2006. The company agreed to pay Mr. Hurst&apos;s legal fees and to pay "substantial" damages, the spokeswoman said. Mr. Murdoch closed the News of the World in 2011, amid revelations that its journalists hacked into the phones and computers of politicians, celebrities and other prominent people. The resulting national scandal forced the company to abandon a bid to buy the 61% of British pay-TV giant Sky PLC that News Corp didn&apos;t already own. Mr. Murdoch since split the company into two: 21st Century Fox Inc., which comprises his TV and movie businesses, and News Corp, which primarily comprises newspapers. Fox is now attempting again to acquire the 61% of Sky it doesn&apos;t own. News Corp publishes several newspapers in the U.S. and U.K., including The Wall Street Journal. Mr. Murdoch and his family are major stakeholders in both Fox and News Corp. Mr. Hurst, who had said News of the World was looking for information related to his work in Northern Ireland and with the Irish Republican Army, sued News Group Newspapers in August 2011.

Assessment of Potential Probiotic Lactobacillus Strains Isolated from Goat Milk Probiotics are a group of microbes that may help directly in enhancing resistance against human intestinal pathogens and in the prevention of gastrointestinal disorders. This study aimed to isolate Lactobacillus spp. from various raw goat milk samples for their probiotic properties. A total of 16 isolates were primarily screened from the collected goat milk samples. Only 3 isolates were found to exhibit remarkable inhibitory activity against multiple pathogenic bacteria. These probiotic strains were analyzed to elucidate their cultural, morphological and biochemical features, and were identified as Lactobacillus bulgaricus, Lactobacillus casei subsp. casei, and Lactobacillus helveticus. The strains were found to exhibit the maximum growth at pH 7 and 37 C temperature for 18 hours of incubation. In addition, they were found to produce antimicrobial metabolites that are inhibitory to pathogens, able to tolerate bile salt at concentration upto 3%, but their inhibitory activity was hampered with trypsin treatment at concentration of 0.5 mg/ml. The strains were found susceptible to erythromycin, chloramphenicol, gentamicin and ciprofloxacin, but resistant against penicillin-G, ampicillin and amoxicillin. The present study thus elucidates that goat milk naturally inhabits Lactobacillus spp. which can be potential candidates in treatment of various infectious diseases in human mainly related with gastrointestinal system. Keyword: Antimicrobial activity, Bacteriocin, Goat milk, Lactobacillus spp., Probiotics

A New Preconditioner for the GeneRank Problem Identifying key genes involved in a particular disease is a very important problem which is considered in biomedical research. GeneRank model is based on the PageRank algorithm that preserves many of its mathematical properties. The model brings together gene expression information with a network structure and ranks genes based on the results of microarray experiments combined with gene expression information, for example from gene annotations (GO). In the present study, we present a new preconditioned conjugate gradient algorithm to solve GeneRank problem and study its properties. Some numerical experiments are given to show the effectiveness of the suggested preconditioner. Introduction. Identifying genes involved in a particular disease is regarded as a great challenge in post-genome medical research. Such identification can provide us with a better understanding of the disease. Furthermore, it is often considered as the first step in finding treatments for it. However, the genetic bases of many multifactorial diseases are still uncertain, and modern technologies usually report hundreds or thousands of genes related to a disease of interest. In this context is where gene-disease prioritization methods are of use. The act of finding the most potentially successful genes among a variety of listed genes has been defined as the gene prioritization problem. Considering rapid growth in biological data sources containing gene-related information such as, for instance, sequence information, microarray expression data, functional annotation data, proteinprotein interaction data, and the biological and medical literature, we can see much interest in recent years in developing bioinformatics approaches that can analyze this data and help with the identification of important genes. The common aim in the present study is to prioritize the genes in a way that those related to the disease under study possibly appear at the top of ranking. In the last decade, several methods have been proposed for ranking or prioritizing genes by relevance to a disease. Some of these methods have been collected at Gene Prioritization Portal: http://homes.esat.kuleuven.be/~bioiuser/gpp/index.php. These methods fall into two broad classes. The first class of the methods mostly uses microarray expression data; these methods focus on identifying genes that are differentially expressed in a disease, and use simple statistical measures such as the t-statistic or related classification methods in machine learning to rank genes based on this property. The second class of methods which is more general, often making use of a variety of data sources; these methods start with some existing knowledge of 'training' genes already known to be related to the disease under study, and directly or indirectly rank the remaining genes based on their similarity to these training genes. There are also some methods that rank or prioritize genes based on their overall likelihood of being involved in some disease in general. Those kinds of methods that purpose to improve an initial ranking obtained from expression data by augmenting it with a network structure derived from other data sources can be related to the methods of second class. For example, the GeneRank algorithm of Morrison et al., is an intuitive change of PageRank algorithm used by the Google search engine that preserves many of its mathematical properties. It combines gene expression information with a network structure derived from gene annotations (gene ontologies (GO)) or expression profile correlations. In the resulting gene ranking algorithm the ranking of genes can be obtained by solving a large linear system of equations. Wu et al. showed that this is equivalent to a symmetric positive definite linear system of equations and analyzed its properties. Then they used the conjugate gradient (CG) algorithm in conjunction with a diagonal scaling to solve the system. Their numerical experiments show that it is the most effective among the tested methods. In this paper, we propose another preconditioner for the GeneRank problem and study its properties. Throughout this paper, we use the following notations, definitions and results. A matrix A is called nonnegative (positive) and is denoted by A ≥ 0 (A > 0) if each entry of A is nonnegative (positive). Similarly, for n-dimensional vectors, by identifying them with n 1 matrices, we can also define x ≥ 0 and x > 0. For a square matrix A, an eigenvalue of A is denoted by (A) and the smallest and largest eigenvalues of A are given by min (A) and max (A), respectively. For any nonsingular The rest of the paper is organized as follows. In section 2 we introduce the GeneRank problem in details. Section 3 is devoted for presenting our preconditioner. In section 4 we present some numerical experiments to show the effectiveness our preconditioner. Finally, concluding remarks are given in section 5. 2. The GeneRank problem formulation. Let the set G = {g 1,..., g n } be n genes in a microarray. Similar to the idea of PageRank, if a gene is connected with many highly ranked genes, it should be highly ranked as well, even if it may have a low rank from the experimental data. In GO, if two genes share at least one annotation with other genes, they are defined to be connected. From this idea, we can build a gene network whose adjacent matrix is W, with entries W ij = 1, g i and g j (i = j) have the same annotation in GO, 0, otherwise. (2.1) In contrast to PageRank, the connections are not directed. Thus, instead of a nonsymmetric hyperlink matrix, GeneRank considers the symmetric adjacency matrix W of the gene network, i.e., W T = W. Let Note that since a gene might not be connected to any of the other genes, W may have zero rows. Now, suppose that the diagonal matrix D is defined by D = diag(d 1,..., d n ), where Then the GeneRank problem can be written as the following large scale nonsymmetric linear system (Morrison et al. ) where I denotes the n n identity matrix. Also is the damping factor with 0 < < 1, and ex = T, with ex i ≥ 0, is the absolute value of the expression change for g i, i = 1, 2,..., n. The solution vector x is called GeneRank vector, and its entries provide information about the significance of a gene. Morrison et al suggested using = 0.5. However, the optimal choice of is still an interesting topic and deserves further study. Note that W is a sparse matrix in general. Morrison et al., used a direct method for the computation of GeneRank vector that is extremely inefficient when the problem is very large or is very close to 1. Yue et al, reformulated the GeneRank model as a linear combination of three parts, and presented an explicit formulation for the GeneRank vector. In Wu et al, the GeneRank problem was rewritten as a large scale eigenvalue problem, and it was solved by some Arnoldi-type algorithms. In, Wu and coworkers proved that the matrix D − W is symmetric positive definite matrix and showed that the nonsymmetric linear system (2.3) can be rewritten as the following symmetric positive definite (SPD) linear system 3). With this modification, methods that are suitable for symmetric systems, can be used for the GeneRank problem. They implemented the Jacobi preconditioner (symmetric diagonal scaling) on D − W. In that case, the preconditioned system is given by x. Wu and coworkers also showed that the eigenvalues of the preconditioned matrix are bounded as follows: These bounds are independent of the size of the matrix, and they only depend on the value of a parameter used in the GeneRank model. They then implemented the classical Chebyshev iteration for the GeneRank problem. It is a polynomial scheme to accelerate the convergence of the stationary iterative method to solve a linear system of equation Ax = b, in which M is a nonsingular matrix. If the matrix I − M −1 A is similar to a symmetric matrix with eigenvalues lying in an interval , then the Chebyshev iteration to solve Ax = b is given by (for more details, see ) where and For the GeneRank problem, it is considered Numerical results presented in show that the number of iterations of the method is usually more than those of the CG method with the diagonal scaling. However, the cost per iteration is much lower and leads to faster convergence in terms of CPU time. 3. Main results. Wu et al. showed that coefficient matrix of linear system (2.4) is symmetric positive definite and therefore we can use the conjugate gradient method ( ) to solve the system. As we know that the convergence rate of the CG method depends on the condition number of the matrix in question, or more generally the distribution of eigenvalues. If the eigenvalue distribution of the preconditioned system is better than that of the original one, the convergence will be accelerated dramatically. For this reason Wu and coworkers applied Jacobi preconditioner on the system (2.4) to accelerate in convergence of the method. In this case, the linear system (2.5) were resulted. Considering (2.6) and (2.7), we can conclude that increasing from 0 to 1 can cause an increase in the ratio of max / min and therefore the coefficient matrix would be increasingly ill-conditioned and the rate of convergence of CG expected to decrease as increases. From now on, for the sake of the simplicity, let J = D − 1 2 W D − 1 2 and S = I − J. It is noted that the matrix S is the coefficient matrix of the system (2.5). We now propose the preconditioner M = I + J for the system (2.5). In this case, the preconditioned system takes the form In the sequel, we investigate the properties of the proposed preconditioner. Proof. Clearly, we have Therefore, According to the relations (2.6) and (2.7) we have 0 < (S ) < 2. Hence, by the latter equation we deduce that 0 < (T ) < 1. Now since T is a symmetric matrix with positive eigenvalues, it follows that T is symmetric positive definite. Obviously, T is a Z-matrix. From Theorem 2.1, S is an M -matrix. Therefore, by Theorem 1.2, there exists a positive vector x such that S x > 0. Hence, T x = (I + J )S x > 0. This shows that the matrix T is an M-matrix. Lemma 3.2. If W = 0, then min (S ) = 1 −. Proof. Let x = (x 1,..., x n ) T, such that otherwise, for i = 1, 2,..., n. Obviously, we have W x = Dx. Now, assuming y = D 1 2 x = 0, we get which is equivalent to S y = (1 − )y. Now by Eq. (2.7), we see that 1 − is the smallest eigenvalue of S. Remark 3.3. In the case that W = 0, we have S = I and there is nothing to investigate. Theorem 3.4. The spectral condition number of T is not greater than that of the matrix S, i.e., Proof. Since both of the matrices S and T are symmetric, it is enough to prove that For every eigenvalue (S ) of S, we have min (S ) ≤ (S ) ≤ max (S ). Then, From Lemma 3.2, we have min (S ) ≤ 1. We now consider two cases, max (S ) ≥ 1 and max (S ) < 1. If max (S ) ≥ 1, then by Eq. (2.6) and Lemma 3.2 we have and since the maximum value of 1 − (1 − (S )) 2 is equal to 1, we get Now, suppose that max (S ) ≤ 1. In this case, it is easy to see that Therefore, the proof of theorem is completed. This theorem shows that the eigenvalues of the matrix T are clustered at least as the matrix S. As we shall see, for the presented numerical experiments the eigenvalues of T are more clustered than those of the matrix S. Numerical experiments. As we mentioned, Wu et al. in successfully employed the Jacobi preconditioner together with the CG algorithm for the solution of the linear system. They have deduced that the CG method in conjunction with the Jacobi preconditioner was faster among the tested methods for each of the presented examples. In this section we compared the numerical results of our preconditioner with those of the Jacobi preconditioner. All the numerical experiments presented in this section were computed in double precision and the algorithms are implemented in MATLAB 7.12.0 and tested on a 64-bit 1.73 GHz intel Q740 core i7 processor and 4GB RAM running windows 7. We use the stopping criteria based on the 1-norm of the residual. That is, we stop iterating as soon as r 1 < tol, where tol is a given tolerance. The initial guess is the null vector. We use two different choices for ex: ex = ( 1 n )e, where e is the vector of all ones, and ex = p, where p is a randomly chosen probability vector -that is, a random vector with entries in. For each adjacency matrix, we use four different values of to form the corresponding GeneRank matrices D − W : = 0.5, 0.75, 0.80, 0.99. The obtained numerical results are presented in some tables. In all the tables, "CG", "PCG", "Chebyshev" and "CG-M " are, respectively, denoted for the CG method implemented for the system (2.4), CG method applied to the system (2.5), Chebyshev iteration and the CG algorithm to the system (2.4) in conjunction with the preconditioner M. Example 4.1. In this example there are three adjacency matrices, w All which is of size 4, 047 4, 047, with 339, 596 nonzero entries, w Up which is of size 2, 392 2, 392, with 128, 468 nonzero entries and w Down which is of size 1, 625 1, 625 with 67, 252 nonzero entries and three expression change vectors expr data, expr dataUp and expr dataDown. These matrices were constructed using the all three sections of the GO, where a link is presented between two genes if they share a GO annotation. Only genes which are up-regulated are included in w Up and only down-regulated in w Down. The data files are available from. The results for the these matrices are given in Tables 4.1, 4.2 and 4.3. In these tables (hereafter for other tables) the number of iterations for the convergence together with the CPU time (in parenthesis) in seconds are given. Here we mention that, in this example, the tolerance tol is taken to be 10 −14. As the numerical experiments show our preconditioner is more effective than the Jacobi preconditioner. It seems that the number of iterations of our method is about half of those of the CG method in conjunction with the Jacobi preconditioner. In addition, as seen our preconditioner outperforms in terms of both number of iterations and CPU times. For more investigation, in Figures 4.1, 4.2 and 4.3 we depict the eigenvalues distribution of S and T for test matrices w Down when = 0.5, w U p when = 0.75 and w All when = 0.9, respectively. As seen the eigenvalues of the T are more clustered than those of S for all the three test matrices. experiments. The first matrix is a SNPa adjacency matrix (single-nucleotide polymorphism matrix). This matrix has n = 152, 520 rows and columns, and is very sparse with only 639,248 nonzero entries. The second type is RENGA adjacency matrix (range-dependent random graph model). In our experiments we set = 0.9 and = 1, the default values in RENGA. Both of these types of matrices are tested in. The results for the SNPa matrix are given in Tables 4.4 and 4.5, and the results for the RENGA matrix with n = 100, 000 and n = 500, 000 are given in Tables 4.6 and 4.7. In this example the tolerance tol is assumed to be 10 −10. As the numerical experiments show the proposed preconditioner is more effective tahn the diagonal scaling and Chebyshev iteration method. Conclusion. In this paper, a preconditioner has been presented for the Gen-eRank problem. Then some of its properties have been given. Finally some numerical experiments have been presented to show the effectiveness of the proposed preconditioner. As seen it does not need any CPU time to set up the preconditioner and the preconditioner is explicitly in hand. Our numerical results show that the proposed preconditioner is more effective than the ones presented in the literature.

The present invention relates generally to a technology of heating related elements to an internal combustion engine and, more particularly, to a control system of a combustion heater for heating constituting elements of the internal combustion engine with heat generated when burning a fuel. In the internal combustion engine, an operability is deteriorated with an increased friction in a low-temperature state, and with this deterioration a fuel efficiency worsens or causing an increase in noises. Further, there arises a problem, wherein a related device operating in connection with the internal combustion engine such as, in a vehicle, for example, a car-room heater with the cooling water of the internal combustion engine serving as a thermal source, is deteriorated in its function when the internal combustion engine is at the low temperature, i.e., when the temperature of the cooling water is low. Accordingly, it is required that the internal combustion engine is provided with a heating device for increasing the temperature of the internal combustion engine itself or a temperature of the related element to the internal combustion engine itself when in a warm-up process after a start-up or when an exothermic quantity of the engine itself is small. This necessity is remarkable especially in a diesel engine with a small exothermic quantity and becomes more remarkable with a tendency of reducing the fuel consumption in recent years. Such being the case, Japanese Patent Laid-Open Publication No. 62-75069 discloses a technology of improving a starting characteristic by speeding up the warm-up of the internal combustion engine. According to this technology, a rise in temperature of the internal combustion engine body is speeded up by speeding up a rise in temperature of the cooling water by providing a combustion heater for heating the cooling water. In this technology, the attention is paid to a temperature when starting the internal combustion engine. In addition to the starting time of the internal combustion engine, however, the temperature of the internal combustion engine becomes comparatively low also when the exothermic quantity of the internal combustion engine itself is small, e.g., when a quantity of fuel consumption in the combustion chamber is small. In such a case also, it is advantageous to the internal combustion engine itself or the related element thereto to provide the combustion heater. Although the combustion of the combustion heater continues during an operation of the internal combustion engine for a short while after the start-up thereof, the exhaust gas from the combustion heater is led into the combustion chamber via the intake passageway. Therefore, the heat of the exhaust gas can be collected in the internal combustion engine, and it follows that an effect of rising the temperature of the internal combustion engine is obtained. In the case of introducing the exhaust gas of the combustion heater into the intake passageway during the operation of the internal combustion engine, however, if the exhaust gas is led into the intake passageway without taking into consideration a combustion state in the combustion chamber of the internal combustion engine, there arises a problem of causing a deterioration of the combustion state in the combustion chamber. For instance, a quantity of the exhaust gas introduced into the combustion chamber from the combustion heater becomes larger, a quantity of fresh air taken in from the atmospheric air becomes smaller. If the quantity of fuel supplied to the combustion chamber is large and a load on the internal combustion engine is high, an air/fuel ratio in the combustion chamber might change to be rich, with the result that more smoke is emitted.

Jamaican sprint star Usain Bolt says he would probably only run the 200 metres if he decides to compete at next year's Commonwealth Games. The six-time Olympic champion hopes to participate in Glasgow, but will discuss it with his coach Glen Mills. 27-year-old Bolt says it's on his to-do list, however the 100-200 double probably would be unlikely. The Jamaican has never won a Commonwealth medal. Bolt came to prominence with victory in the 100m, 200m and 4x100m relay at the 2008 Beijing Olympics and did the same in London four years later. He now aims to complete the 'treble treble' by winning all three disciplines at the 2016 Games in Rio to cement his legend status. Bolt is considering putting plans to end his career after the next Olympics on hold to run at the 2017 World Championships in London.

(CNN) - To call Mia Love a minority is an understatement. She’s a black woman who won an upset primary race to become the Republican candidate in Utah’s 4th Congressional District. If elected, she’d be the first black Republican congresswoman in the House of Representatives. Love, who has attracted lots of national Republican support, also stands out because of her religion: She’s a Mormon. The politician is a poster child for the Church of Jesus Christ of Latter-day Saints’ campaign to present a more diverse face to a historically very white church. Most Muslims in several key Middle Eastern and Asian countries hold negative views of the terrorist network al Qaeda a year after U.S. forces killed its leader Osama bin Laden, according to a recent survey. The Sikh Coalition unveils new app to combat racial profiling. The Sikh Coalition, a civil rights advocacy group, on Monday released a mobile application on iPhones and Android phones giving passengers who feel they've been racially or religiously profiled a way to speak out against screeners with the Transportation Security Administration. (CNN) - Most Muslims in several key Middle Eastern and Asian countries hold negative views of the terrorist network al Qaeda a year after U.S. forces killed its leader Osama bin Laden, according to a recent survey. The poll by the Pew Research Center's Global Attitudes Project, released Monday, found that a high proportion - between 71% and 98% - of Muslims questioned in Egypt, Jordan, Turkey and Lebanon viewed al Qaeda in an unfavorable way.

1. Field of the Invention The present invention relates to an inflator that provides inflation fluid to inflate an inflatable vehicle occupant protection device and, more specifically, to an initiator retainer for an air bag inflator. 2. Description of Related Art An inflatable vehicle occupant protection device, such as an air bag, is inflated upon the occurrence of a vehicle condition requiring inflation of the air bag. When inflation is required, an inflator is actuated to provide inflation fluid, which inflates the air bag into the vehicle occupant compartment. The inflator includes a container defining an inflation fluid pressure chamber with an outlet passage. A rupturable closure member is fixed to the container to block flow of inflation fluid through the outlet passage. The inflator further includes an electrically actuatable initiator which, when actuated, causes the closure member to rupture so that inflation fluid in the pressure chamber can flow from the inflator. In some applications, it is desirable that the inflation fluid exit the inflator in a direction parallel to a longitudinal axis of the inflator. The present invention is an inflator for inflating an inflatable vehicle occupant protection device. The inflator comprises a container in which inflation fluid is stored under pressure. The container has an opening through which inflation fluid flows in a given direction from the container. A rupturable closure member is fixed to the container and blocks flow of inflation fluid through the opening. The inflator includes an initiator for, when actuated, rupturing the closure member to enable inflation fluid to flow from the container through the opening. The inflator also includes a retainer for retaining the initiator on the container. The retainer comprises at least one part having a passage for directing gas that flows from the container in the given direction. The inflator still further includes a support for the rupturable closure member. The rupturable closure member has a first portion that is deformed into engagement with the support by the pressure of the inflation fluid in the container. The support transmitting force from the closure member to the retainer.

Gastroenteritis at a University in Texas: An Epidemiologic Case Study This CD-ROM is an important addition to case exercises in field epidemiology that serve to educate when actual participation in a field investigation is not possible or practical. The authors have prepared a case exercise based on an actual field investigation with real data that have been put together in a meaningful and effective way. The use of an epidemic of gastroenteritis is a cutting-edge element, since foodborne disease is a major public health problem today. The outbreak occurs on a college campus, which lends an air of verisimilitude, and the causative agent, norovirus, is a genuine public health threat. This reviewer had a number of specific editorial recommendations for the authors that could enhance forthcoming versions. These suggestions included inserting a case definition in the investigation outline; adding the role of the state laboratory; consistently labeling outbreak, epidemic, and epidemic curve throughout the program; clarifying the rationale for limiting the outbreak to the university; further refining methods for the study and controls; using 22 tables to illustrate epidemiologic ratios; and expanding the employee training plan. Overall, these types of training aids are needed as we attempt to further expose public health workers to field investigations so that they can conduct investigations effectively. The reference to additional educational material throughout the steps is a well-conceived and appropriate aspect of the investigation. The narrative information, questions, and explanations are appropriate and flow smoothly.

Analysis of economic trends in overseas markets for Indian tea and coffee Since world market economies are dynamic, the responsiveness of export quantities to changes in international prices is of direct relevance in international economics. India has 12% share of world tea exports and is the seventh largest coffee producer in the world (20132014). This study analyses the exports and price growth rates, elasticity, instability and major global markets for Indian tea and coffee. The study was based on 24 years agricultural time-series data from 19901991 to 20122013. The study found that elasticity for coffee and tea was 61.4 and 13.5%, respectively. The destinations which account for the major share of exports for Indian tea are developed countries including Australia (18.7%), Russia (17.3%), and the United States (15.9%) and for coffee are Italy (28.7%), Germany (13.8%), and Belgium (7.6%). Based on an analysis of individual country elasticity and growth rates, India has comparative advantage to export to the Middle East, Europe, the United States, and Australia.

Improvement of High Strength Automotive Steels Wettability Properties Using CO2 Laser Surface Treatment As a result of stricter environmental and safety standards, vehicle manufacturers have to reduce the weight of the vehicles, because 10% weight loss cause 8-10% reduction of fuel consumption. To reduce cars weight and increase safety, vehicle manufacturers use high-strength steels. Further weight reduction can be achieved by using corresponding bonding technology (soldering, sticking) and optimizing these technologies can increase the strength of the joints. According to literature research, the improvement of interface properties has a large effect on bonding technologies. In order to improve interface properties, we can use multiple surface treatments. In our research we investigate the effects of CO2 laser surface treatment on high strength steels, because CO2 lasers are often used in the vehicle industry. In order to detect the effect of surface treatment, we investigate the wettability of the treated and untreated steels. In our research we measure the surface tension of treated and untreated steels. Our main goal is to improve wettability properties thus the bonding technology. In our research we used DP 600 high strength steel sheet with the thickness of 1 mm. We cut the steel sheet to 25mm wide and 55mm long workpieces. Before the surface treatment, the workpieces had to be cleaned and degreased using methanol. We searched for parameters that do not cause any visible changes on the surfaces. Among the parameters of the treatment we were able to change the output power of the laser. We used contact angle measurement to examine the wettability.

Liquid biopsy of extracellular vesicle biomarkers for prostate cancer personalized treatment decision Liquid biopsy of tumor-derived extracellular vesicles (EVs) has great potential as a biomarker source for prostate cancer (CaP) early diagnosis and predicting the stages of cancer. The contents of EVs play an important role in intercellular communication and have specific expression in blood and urine samples from CaP patients. Powered by high-throughput, next-generation sequencing and proteomic technologies, novel EV biomarkers are easily detected in a non-invasive manner in different stages of CaP patients. These identified potential biomarkers can be further validated with a large sample size, machine learning model, and other different methods to improve the sensitivity and specificity of CaP diagnosis. The EV-based liquid biopsy is a novel and less-invasive alternative to surgical biopsies which would enable clinicians to potentially discover a whole picture of tumor through a simple blood or urine sample. In summary, this approach holds promise for developing personalized medicine to guide treatment decisions precisely for CaP patients.

According to a classified FBI counterterrorism policy guide obtained by The Intercept, "white supremacists and other domestic extremists" have been joining law enforcement agencies across the United States. "Domestic terrorism investigations focused on militia extremists, white supremacist extremists, and sovereign citizen extremists often have identified active links to law enforcement officers,” the FBI's guide states. Detailing the ways the FBI places individuals on a terrorism watch list, the Known or Suspected Terrorism File. Advertisement: Despite the evident threat posed by these right-wing extremists, law enforcement has failed to acknowledge proper ways to combat, or even address the systemic issue. "No centralized recruitment process or set of national standards exists for the 18,000 law enforcement agencies in the United States, many of which have deep historical connections to racist ideologies. As a result, state and local police as well as sheriff’s departments present ample opportunities for white supremacists and other right-wing extremists looking to expand their power base," The Intercept wrote. In October 2006, the FBI released an internal assessment noting that white supremacy groups have a history intending to infiltrate law enforcement agencies. The memo contained many redactions, but noted "ghost skins," or individuals who mask their personal beliefs in order to integrate with society, while still secretly advancing their ideologies. "In at least one case, the FBI learned of a skinhead group encouraging ghost skins to seek employment with law enforcement agencies in order to warn crews of any investigations," The Intercept reported. The report came about after widespread instances of abuse, including a local sheriff's department that formed a neo-Nazi gang that consistently terrorized black and latino communities, and a police detective in Chicago rumored to be a KKK member that tortured at least 120 black men throughout his career, to name a few. After Barack Obama took office, an intelligence study conducted by the Department of Homeland Security, with the partnership of the FBI, warned of a "resurgence" in right-wing extremism. The report stated, "lone wolves and small terrorist cells embracing violent right-wing extremist ideology are the most dangerous domestic terrorism threat in the United States." Advertisement: These conclusions resulted in unrest from many conservatives who felt as if they were painted in a bad light, and presumed dangerous, by association. Due to the criticism, the report was condemned by then DHS Secretary, Janet Napolitano, who also issued an apology to military veterans. Despite re-establishing the Domestic Terrorism Task Force in 2014, the majority of the government's efforts to thwart extremism was focused on foreign groups, rather than domestic. "Critics fear that the backlash following the 2009 DHS report hindered further action against the growing white supremacist threat, and that it was largely ignored because the issue was so politically controversial," The Intercept wrote. The Intercept cited sociologist Pete Simi, who has studied for decades the rapid increase in white supremacists in both law enforcement and the military. Simi conducted a study finding at least 31 percent of individuals indicted for right wing extremist activities, to have military experience.

Field Experiments in the Global South: Assessing Risks, Localizing Benefits, and Addressing PositionalityADDENDUM Randomized controlled trials (RCTs) have emerged as a leading methodological tool to strengthen causal inference in the social sciences. Yet RCTs carry significant risks for everyone involved, from participants to researchers themselves, especially in the Global South. In this article, we explore how researchers identities and power influence the conduct of research and their positionality within the research contextsespecially when conducting RCTs in the Global South. Our goal is to center local contexts and demands at each stage of the research process. Overall, we argue that centering local contexts, stakeholders, and demands at each stage of the research process is key to ensuring that RCTs in the Global South are ethically sound and generate insights that can serve the communities they investigate.

Chinese web censors eradicate SCMP presence in mainland China’s internet censors have shut down the South China Morning Post’s social media accounts, effectively eradicating the Hong Kong English-language newspaper’s online presence in the mainland. The SCMP’s microblogs on Sina Weibo and Tencent Weibo and its WeChat page used to post updates from the paper’s Chinese-language website, SCMPChinese.com (nanzao.com), which is blocked in the mainland. But the newspaper’s official Sina Weibo page now leads viewers to an error message that reads, “Sorry, there is something wrong with the account you are currently trying to access, and it is temporarily inaccessible.” A similar error message can be seen on what used to be SCMP’s Tencent Weibo page. At SCMP’s official WeChat account, an empty shell is all that remains. All previous posts published on the page have been deleted. If past history is any indication, the shutdown doesn’t appear temporary, and there is little chance that the accounts will be restored, Shanghai-based English-language website Shanghaiist reported Wednesday. Launched in April 2013, SCMP’s Chinese portal, with its mix of translations from the English-language paper and original Chinese-language content, was hailed by the publisher as a major push into the digital space and into the wider Chinese market. The US$266 million acquisition of the newspaper assets by Jack Ma Yun’s Alibaba Group Holding Ltd. in December has stoked fears that the SCMP’s independence would continue to be compromised, as Ma’s ties to the governing elite in Beijing are well-known, Shanghaiist said. Following President Xi Jinping’s recent high-profile visits to state media outlets and his declaration that media outlets must be “surnamed Party” so they can achieve “correct guidance of public opinion” by “singing the main theme, transmitting positive energy”, influential weibo celebrity and former property tycoon Ren Zhiqiang wrote on his personal account that state media should serve the public, not the Communist Party. One week later, China’s top internet watchdog shut down his social media accounts, where he had accumulated more than 37 million followers. Like Ren, the SCMP was a powerful force on weibo. In March 2015, it was named one of the most influential Hong Kong media sources on weibo for the second consecutive year in The Star of Weibo awards, organized annually by Sina Weibo. Less than a year later, the SCMP appears to have lost its more than 500,000 followers on weibo overnight. With a dwindling pool of English-language readers at home and its ability to penetrate mainland China now severely limited, the future looks bleak for the Post, Shanghaiist commented. – Contact us at [email protected] FL

Screenshot by Chris Matyszczyk/CNET When it comes to image, Wall Street's remains somewhere below that of Lance Armstrong and Anthony Weiner. You'd think, therefore, that Wall Street types might have some sensitivity to how they're seen in the outside world. You'd also think that mud is the perfect gravy for a french fry. At least that is the view of Brendan O'Connor, the former cook at the well-known four-wheeled New York food provider Milk Truck. For he was cheesed off when some people from Glass, Lewis & Co came to his truck, ordered around $170 worth of food, and then decided to leave no tip. A crime of omission on commission is not to be taken lightly in the food business -- as, one supposes, it wouldn't be appreciated on Wall Street. My own local restaurant has a lady who comes in and tips a pittance. She is not well-regarded. Perhaps you might think that food trucks don't deserve tips. But on a $170 order, might it not have been a decent gesture? So O'Connor took to Twitter and made the following statement: "Shout out to the good people of Glass, Lewis & Co. for placing a $170 order and not leaving a tip. @glasslewis." It's possible that the "@glasslewis" part alerted the Wall Streeters to O'Connor's grief. So the company contacted Milk Truck and, suitably humbled, offered to pay $30 as a tip. No, I don't have that quite right. As O'Connor himself wrote in the Awl, the company allegedly contacted O'Connor's boss and then he was fired. Indeed, as O'Connor wrote in the Awl: "Obviously I knew it was a possibility that I'd get fired. I guess I had hoped that the owner would have my back if they complained, but that was a miscalculation." Perhaps you can understand that from the Milk Truck owner's point of view. He has a small business to run. O'Connor admits he wasn't the finest of employees. But what about the folks at Glass Lewis? Instead of reacting to the tweet by complaining, what if they'd slapped their foreheads in a public place and returned O'Connor's tweet like so: "We forgot the tip? Shame on us. We've been so bloody busy. We're on our way down to make amends"? Wouldn't the company have come off rather better than it looks now? Currently, the #glasslewis hashtag is enjoying some painful commentary. I have contacted Glass Lewis to ask for the company's view and will update, should I hear. When someone offers you brusque bluster on Twitter, often they might be entirely full of cat food and last night's beer. It's sometimes even worth retweeting their nonsense, which can be decorated with badly spelled curses. Just occasionally, though, isn't it worth considering whether the tweet might have merit? O'Connor's tweet was mere sarcasm, the sort that you'd hear on Wall Street every day. Doesn't Glass Lewis, a self-proclaimed "Global Governance Leader," believe in tipping on $170 worth of food?

Successful management of severe allergic reactions to platelet transfusion with omalizumab Abstract Rationale: An allergic transfusion reaction is a common side effect of transfusions of red blood cells. Using washed red blood cells is the most effective method for preventing such a reaction. However, the availability of other washed transfusion components, including platelets, is limited. Patient concerns: A 69-year-old patient with acute myeloid leukemia progressed from myelodysplastic syndrome and was treated with azacitidine. She experienced a minor reaction to platelet transfusion that initially responded to the administration of corticosteroids and antihistamines. However, she worsened even after subsequent preventive treatments and was referred to the emergency department due to anaphylaxis. The patient developed hypotension, chest pain, and dyspnea 10 minutes after the initiation of platelet transfusion. Diagnoses: She was diagnosed with platelet-induced anaphylaxis. Interventions: In an attempt to prevent anaphylaxis, 150mg of omalizumab was prescribed 1week prior to transfusion. However, she experienced anaphylaxis again and was administered intramuscular epinephrine. For the following transfusion, we treated her with a 300mg dose of omalizumab 24hours before the transfusion. Outcomes: She tolerated well and continued to receive further chemotherapy and platelet transfusion with premedication. Lessons: This case suggests that omalizumab is a good candidate for the management of severe allergic transfusion reactions. Introduction Allergic transfusion reactions (ATRs) are common side effects of transfusion. Many different blood components can induce such reactions, and they are more common in cases of platelet or plasma transfusion due to the activation of leukocytes and the accumulation of cytokines when these transfused products are stored at room temperature. The incidence of ATRs is 0.3% to 6% for platelet transfusion and 1% to 3% for plasma transfusion. The etiology of ATRs remains unclear. However, both the allergic characteristics of the patient receiving the transfusion and the cellular and plasma components of the transfusion products can contribute to the development of ATRs, specifically type 1 hypersensitivity reactions in which immunoglobulin E (IgE) interacts with components in the donor plasma. Manifestations vary from mild allergic reactions to severe anaphylaxis. Specific medical conditions, such as IgA or haptoglobin deficiency, can make patients more susceptible to anaphylaxis. Mild reactions can be managed with corticosteroids or antihistamines, but severe anaphylaxis must be treated with prompt injection of epinephrine. To prevent ATR recurrence, pretreatment with antihistamines and systemic corticosteroids can be used in mild cases; further, the use of washed blood components has been shown to be helpful in some cases. However, there are no definitive guidelines to prevent the recurrence of severe ATRs, especially those induced by platelet transfusion. Omalizumab is a recombinant, humanized, monoclonal anti-IgE antibody that binds to circulating IgE molecules and prevents Editor: Maya Saranathan. The ethical committee waived the requirement for obtaining approval from the institutional review board for case reports. The patient provided informed consent for the publication of the case. The authors have no funding and conflicts of interest to disclose. The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. OPEN IgE binding to high-and low-affinity IgE receptors (FceRI and FceRII) on effector cells. It is widely used for the treatment of severe asthma and refractory chronic urticaria. In limited cases, omalizumab has been used to manage anaphylaxis. We report a case of platelet-induced anaphylaxis that was successfully managed with omalizumab. Case presentation A previously healthy 69-year-old female patient was diagnosed with myelodysplastic syndrome and refractory anemia with excess blasts-1 in June 2016. Her physical examination revealed no specific findings at the time of diagnosis, but the initial complete blood count was indicative of pancytopenia (white blood cell count, 1040/mL; hemoglobin level, 8 g/dL; and platelet count, 28,000/mL). She received azacitidine chemotherapy (75 mg/m 2 ; body weight, 61 kg). After 3 rounds of chemotherapy, the patient achieved complete remission, as confirmed by bone marrow biopsy, and continued the treatment course. However, the pancytopenia worsened despite 32 rounds of azacitidine chemotherapy; a complete blood count on March 13, 2020, revealed a white blood cell count of 590/mL, hemoglobin of 7.1 g/ dL, platelet count of 42,000/mL, and absolute neutrophil count of 136/mL. Repeated bone marrow biopsy revealed progression to acute myeloid leukemia with myelodysplasia-related changes. The patient occasionally had adverse ATRs during transfusion. Initially, mild ATRs occurred after the patient was transfused with red blood cells (RBCs), and they were successfully prevented using washed RBCs. The patient also experienced mild ATRs after platelet transfusion, which were prevented using irradiated leukocyte-depleted platelets and pretreatment with antihistamines (4 mg chlorpheniramine) and systemic corticosteroids (100 mg hydrocortisone) until November 29, 2019. When she was transfused with irradiated leukocyte-depleted platelets on February 11, 2020, she experienced shortness of breath, abdominal cramping, and nausea but no skin manifestations. The patient was referred to the emergency department. Her vital signs were stable without desaturation or hypotension, and her symptoms were relieved after treatment with antihistamines and systemic corticosteroids. Baseline IgA levels were normal at 153.97 mg/dL (normal range: 70-400 mg/dL), serum tryptase levels were normal at 1.4 mg/L (normal range: < 11 mg/L) measured 2 hours after anaphylaxis, and total IgE levels were 62.6 kU/L (normal range: < 100 kU/L). As ATRs were successfully prevented using antihistamines and systemic corticosteroids previously, we planned to increase the dosage and duration of these medications as follows: intravenous administration of 40 mg methylprednisolone at 1, 7, and 13 hours prior to transfusion and 4 mg chlorpheniramine 1 hour prior to transfusion. However, 3 minutes after the initiation of platelet transfusion, the patient complained of chest tightness, and her blood pressure dropped rapidly from 140/90 to 105/75 mm Hg. The patient recovered from shock after intramuscular epinephrine injection. Therefore, we decided to pretreat the patient with omalizumab, an anti-IgE monoclonal antibody, before the next transfusion. Initially, a dose of 150 mg omalizumab was administered to the patient 1 week before the platelet transfusion in addition to systemic corticosteroids and antihistamines administered 13 hours prior to the transfusion. However, chest tightness, abdominal cramps, and shortness of breath with desaturation occurred within 10 minutes after platelet transfusion. For the next transfusion, we administered 300 mg omalizumab 24 hours before transfusion, resulting in the successful completion of the transfusion without recurrent anaphylaxis. Since that time, the patient has tolerated platelet transfusion accompanied by pretreatment with 300 mg omalizumab, systemic corticosteroids, and antihistamines (Fig. 1). Discussion ATRs occur commonly during transfusion. They have been classified into different subtypes, including febrile nonhemolytic reactions, allergic reactions, delayed and hyper-hemolytic reactions, transfusion-related graft-versus-host disease, transfusion-related acute lung injury, and massive transfusion-associated reactions. The management of these different types depends on the specific type of ATR and the blood components transfused. Anaphylaxis is a rare manifestation, with an incidence of < 1%. The use of antihistamines and glucocorticoids is the current gold standard, especially in patients who receive multiple units of platelets (from one or multiple donors), meaning that they are more likely to be exposed to allogenic antibodies and develop reactions easily. Therefore, the removal of plasma components may help prevent ATR. There is some evidence that concentrated apheresis can reduce the incidence of ATRs. A washed product is a more effective way to remove plasma under 10%; however, the limited availability of manufacturing is challengeable. We successfully treated our patient with Several mechanisms account for anaphylaxis, one of which is IgA deficiency. According to a previous report by Shi et al, a patient exhibited borderline levels of IgA; however, due to limitations, no additional studies were performed. Our patient had completely normal laboratory results, including initial levels of IgA, tryptase, and other immunoglobulins. The patient initially experienced febrile nonhemolytic transfusion reactions, which were later followed by anaphylaxis. In our case, we did not measure other serum proteins or autoantibodies; therefore, it was difficult to clarify the origin of the ATRs. We hypothesized that our patient produced IgE in response to an unknown plasma protein after receiving transfusion products at certain times. According to one report, multiple components in the plasma, such as complement or haptoglobin, are regarded as candidate triggers of ATRs. Omalizumab is an anti-IgE antibody that binds to free IgE in human plasma or interstitial fluid and reduces sensitivity to allergens. In addition to its antiallergic effects in steroidrefractory asthma, some studies have found that omalizumab can stabilize mast cells and act as an immune-stabilizer in nonallergic diseases. Therefore, omalizumab has been approved for the treatment of chronic urticaria, which is influenced by both allergic and nonallergic mechanisms. It has also been used offlabel for the prevention of anaphylaxis during specific allergen immunotherapy, anaphylaxis during mastocytosis, idiopathic anaphylaxis, and transfusion-induced anaphylaxis. The optimal dose of omalizumab for allergic asthma is 75 to 325 mg subcutaneously injected every 2 or 4 weeks; a dose of 300 mg every 4 weeks is commonly used for chronic idiopathic urticaria. However, there is no recommended dose for off-label use. Some case reports of systemic mastocytosis show that multiple doses of 150 or 300 mg omalizumab may effectively manage anaphylaxis. In a previous case of platelet-induced anaphylaxis in a patient with IgA deficiency by Shi et al, the patient was pretreated with 150 mg of omalizumab and successfully tolerated platelet transfusion. However, in our case, platelet-induced anaphylaxis was not prevented by pretreatment with 150 mg omalizumab 7 days prior to transfusion. The patient was then treated with 300 mg omalizumab 24 hours before transfusion, which successfully prevented platelet-induced anaphylaxis. The successful prevention of platelet-induced anaphylaxis may be more likely due to the increased dose of omalizumab rather than to the timing of the injection, considering that peak concentrations of omalizumab are reached 7 to 8 days after injection. In conclusion, omalizumab is a promising candidate for the prevention of transfusion-induced anaphylaxis. Further studies are warranted to optimize its dose and efficacy, which will depend on baseline IgE levels as well as the time of drug administration.

The Importance of Relationships in Education: Reflections of Current Educators The purpose of this study was to seek to understand and describe current educators perceptions of building relationships in education. Thirty-five K-12 educators completed reflective writing responses as part of this phenomenological study. The thematic analysis constructed four themes: (a) students are motivated to learn when strong relationships are formed, (b) if they dont trust you and dont think you care, learning decreases, (c) teachers had a great impact on me that I admire deeply-these teachers are the reason I went into education, and (d) they can always google the content. The findings highlight the significant impact of relationships.

Mathematical model of negative control of E-coli bacteria A mathematical model for the negative control of the lac operon is presented. This model consists of coupled nonlinear differential equations representing the dynamic behaviors of the repressor-inducer components of negative control components of the system. To eliminate the dependency of the magnitude on the system of units of measurements, a dimensionless derivation for the operon system was employed to produce singularly perturbed models with fast and slow time-scale models. The fast model represents the dynamical behavior of the operator. Whereas the slow model represents the dynamical behaviors of the inducer and the repressor. It is shown that by applying the singular perturbation theory we can study the approximate behavior of the free operator by solving one equation instead of three coupled nonlinear differential equations. The behavior of the system can be described as a rapid on-off switch of structural gene transcription. This is consistent with the requirement of the bacterial cell to quickly respond to nutrient fluctuations.

Torque ripple minimization of Matrix Converter-fed PMSM drives using Model Predictive Torque Control A Model Predictive -Direct Torque Control (MP-DTC) technique is proposed for Matrix Converter (MC)-based Permanent Magnet Synchronous Motor (PMSM) drive system, which reduces the torque ripples, does not need the duty cycle calculation, and ensures the fixed switching frequency. The predictive model of PMSM and MC is realized by means of State model. Then, the advanced MC-fed model predictive DTC algorithm is implemented based on cost function evaluation. The simulation results exhibit remarkable torque ripple reduction with the help of Model Predictive Control (MPC). As a result, the proposed strategy is proved to be effective in minimizing the torque ripples for MC-based PMSM drives.

BONN, Germany (Reuters) - Germany’s foreign minister on Thursday warned that building more Israeli settlements in the Palestinian territories could end the prospect of a two-state solution and fuel conflict in the region. Sigmar Gabriel’s comments came as conflicting statements by the new U.S. administration threw off European allies who had hoped to get some clarity from Washington following U.S. President Donald Trump’s apparent shift in policy on Wednesday regarding the Middle East peace process. “We are concerned that unlimited construction of settlements will ... make a two-state solution impossible and could increase the risks of conflicts in the Middle East, including possible war,” Gabriel told reporters, showing Berlin’s growing frustration about settlement activity in the Israeli-occupied West Bank. A vote by the Israeli Knesset to “legalize” settlements banned under international law further complicated the situation, Gabriel said during a news conference at a G20 foreign ministers meeting. Trump on Wednesday dropped a U.S. commitment to a two-state solution to the Israeli-Palestinian conflict, abandoning a major pillar of U.S. Middle East policy. But on Thursday, U.S. Ambassador to the United Nations Nikki Haley said it would be wrong to say that Washington no longer supported a two-state solution. French Foreign Minister Jean-Marc Ayrault told reporters after a meeting with U.S. Secretary of State Rex Tillerson that the U.S. position on the Israeli-Palestinian dossier was “very confused and worrying”. Gabriel, who is due to meet with Tillerson later on Thursday, said Germany would continue to advocate a two-state solution for the Israeli-Palestinian conflict, calling it “the only realistic option to reduce conflict in the region and prevent the emergency of a new war”. Germany’s concerns about settlements have already derailed a meeting planned between the German and Israeli governments in May, with one senior German official saying ties between the two countries had been “completely pared back.” German governments have made strong relations with Israel a top priority ever since World War Two, going to great lengths to make amends for the killing of six million Jews by the Nazis. But relations have grown tense in recent years as Germany questioned Netanyahu’s commitment to a two-state solution with the Palestinians. Israel cites biblical, historical and political connections to the West Bank and East Jerusalem, which it captured in the 1967 Middle East war and where some 2.6 million Palestinians live. It has built about 120 settlements in the West Bank. About 350,000 settlers live there, with a further 200,000 in East Jerusalem. Settlement building activity has increased sharply since Trump took office last month, with Netanyahu approving construction of 6,000 settler homes in the two areas - moves widely condemned internationally, but not by the White House.

A Sacramento police cruiser's dash cam recorded officers saying things like "F— this guy," and "I'm going to hit him," as they appear to try to run over a mentally ill homeless man, Joseph Mann, 51, whom they were sent to confront. (City of Sacremento) “F— this guy,” the officer says before aiming his police cruiser at the mentally ill homeless man that he and his partner had been sent out to confront. “I”m going to hit him.” “Okay, go for it. Go for it,” his partner responds, his voice recorded on the Sacramento police cruiser’s dash cam. Twice, the man dodges their accelerating cruiser. In the second attempt, he leaps into a median, barely avoiding the vehicle. But Joseph Mann, 51, could not escape the volley of bullets that followed moments later. Mann died in the street shortly after that July 11 shooting. Mann’s family has accused officers Randy Lozoya and John Tennis of escalating the situation and showing no regard for his life, even before they jumped out of the car, fired 18 bullets and shot Mann 14 times. The family wants the officers charged with murder. They say the video, the audio of which was enhanced by the Sacramento Bee and published Friday, is damning. “They are officers that shouldn’t be in uniform,” Robert Mann, Joseph Mann’s brother, told the newspaper. “If this is their state of mind when they go to work, this doesn’t serve anyone well.” Robert Mann, right, addresses community members at Allen Chapel AME Church in Sacramento, accompanied by his attorney, John Burris. (Darcy Costello/Associated Press) On Monday, the family asked for a U.S. Justice Department investigation into the Sacramento Police Department, the newspaper reported. “This is murder and a reckless disregard for human life,” the family’s attorney, John Burris, said at a Monday press conference, according to the paper. The attempts to run down Mann “epitomized the lawlessness of these officers.” The Mann case was not the first time Tennis has killed a man in the line of duty, Burris said. In 1997, Tennis placed Albert Thiel, a 35-year-old black man, in a chokehold. Thiel died during the violent struggle. The deaths of Thiel and now Mann are a sign of larger problems in the department, according to Burris. “There are policies and procedures in place that really need to be checked,” he said. “There are issues here about use of the car, how do you treat the mentally impaired, that it’s important that the DOJ … takes a look at.” Lozoya and Tennis have been placed on desk duty. Mann’s family has filed a federal lawsuit and a claim against the city, which they plan to update to include the attempts to hit Mann with the car, the newspaper reported. Police released the dash-cam video two weeks ago, after resisting for two months following the fatal confrontation. On July 11, they were called to a brightly lit street in California’s capital by neighbors who reported a man standing in the street, waving a knife. That was Joseph Mann, who was in the midst of a mental health crisis, according to court documents filed by his family. They say he was “doing karate moves and zigzagging back and forth across the street as he tried to walk away from the Officers.” Mann also had methamphetamine in his system, according to police. And he wasn’t cooperating with the officers who arrived. A lawsuit filed by Mann’s family said he was “displaying obvious signs of mental distress.” It faults the department for not trying to de-escalate the situation. “Inexplicably, the Officers failed to contact any properly trained mental health counselors or make any attempt to use less than lethal force and ignored the established police protocols to make attempts to de-escalate the situation,” the lawsuit said. [Philadelphia gunman, ‘driven by hatred,’ ambushed officer, then went on deadly rampage] “Instead, Officers left their (positions) of safety, cornered Mr. Mann against a (fence/wall) and opened fire on him.” The shooting happened as officers across the nation have been embroiled in a debate about whether they are too quick to use lethal force, especially against minorities. Police have killed 719 people so far this year, according to a Washington Post examination of fatal force. Like Mann, approximately 1 in 4 of those killed were reported to have some form of mental illness. A few days before Mann was shot to death, a gunman opened fire on police officers at a Black Lives Matter protest in Dallas, killing five and increasing tensions between police and the communities they serve. Mann’s shooting sparked protests in Sacramento as residents demanded the release of the dash-cam videos. For months, police claimed the videos were not public record because they were part of the investigative file. But two weeks ago, amid mounting public and political pressure, officials relented, releasing video from three dashboard cameras and one from a nearby business. The most incendiary one has been the video shot through the windshield of Lozoya and Tennis’s police cruiser. It shows the series of close calls as the officers tried to hit Mann. After the second attempt, the vehicle is parked and two officers can be seen running in Mann’s direction. Then, shots are fired. This post has been updated. Read more: After shootings, police fear being ‘a public target’ — even at lunch One-third of officers killed responding to calls were alone, new study of police deaths finds The Charlotte police shooting that hasn’t gone away

Success in Theory of Mind Peter wants to get the beer that he left in the refrigerator. Predicting Peters behaviour correctly is usually an easy matter, but understanding how people correctly predict his behaviour with ease is a much more difficult task. Thirty years of research on theory of mind has focused on the interesting few cases in which people fail to reason about mental states correctly, however it is perhaps more interesting to explore the common, reliable cases of successful theory of mind reasoning. This symposium presents research exploring successful instances of theory of mind reasoning using a variety of experimental approaches, and examines the ability to succeed consistently across the lifespan, with results from toddlers, preschoolers, young children, and adults. Important conclusions are drawn from the presented research, which includes the first evidence that children as young as 2.5 years of age can succeed on explicit false belief tasks (Scott & Baillargeon), the most direct behavioral evidence to date for inhibitory processing in successful behavior prediction based on false belief and avoidance desire in preschoolers and young children (Petrashek & Friedman), and, in adults, evidence from a probabilistic modeling approach to theory of mind and social learning development with extensions to pragmatic language usage and natural pedagogy (Goodman).

A Novel DFT-based Digital Watermarking System for Images A novel design of a DFT-based digital watermarking system for images is proposed in this paper. The system provides images copyright protection which can be done by adding invisible watermark into the digital image. First, the original image is decomposed into DFT coefficients using a fast Fourier transform. For minimal loss in image fidelity, the watermark is embedded in those DFT coefficients with highest magnitudes except for those in the lowest one. The watermark detection is achieved without use of the original image by computing a similarity measure between the input watermark and the DFT coefficients of the attack image. Experimental results show that the proposed method is robust against the common image/signal processing operations

Autonomous and Operator-Assisted Electric Rope Shovel Performance Study Automation has been changing the mining industry for the past two decades. Material handling is a critical task in a mining operation, and truck-shovel handling systems are the primary method for surface mining. Mines have deployed autonomous trucks, and their positive impact on both production and safety has been reported. This paper aims to study the extent to which autonomous and operator-assisted loading units could improve different aspects of a mining operation. Four different levels of automation ranging from operator-assisted swing and return to fully autonomous for a shovel were considered. A discrete event simulation model was developed and verified using detailed data from a shovel monitoring system. Later, the developed model was deployed to assess how each of the proposed technologies could improve productivity and efficiency. Results show that up to a 41% increase in production can be achieved. Both mining companies and equipment manufacturers can use the methodology and results of this study for future decision-making and product development.

Moving Beyond Chalk and Talk: Using Problem-Based-Learning In A Research Methods Course Sequence The average adult can concentrate for only about eight to ten minutes during an hour-long lecture. Thus, students' ability to absorb information may be seriously impeded if we college professors talk nonstop. One alternative to the traditional "chalk and talk" instructional method is problem-based learning (PBL) - an instructional approach using real world problems as a format for students to acquire critical thinking, problem solving and group interaction skills. We describe how we transformed a two-course sequence in research methods into a problem-based-learning format. Student-reported benefits of the PBL approach include the need for higher-order thinking, improved group interaction skills, relevance of course material to real world situations, higher motivation and an overall higher level of class enjoyment.

ZoneAlarm, which has been around since the late 1990s, is well known for its free firewall; more recently, it has been marketing a full security suite. With ZoneAlarm Security Suite 2010, Check Point Software Technologies Inc. (which purchased the product from Zone Labs in 2004) has integrated its firewall and spyware-prevention products into a suite that incorporates Kaspersky’s virus-scanning engine to create a full array of anti-malware, anti-intrusion capabilities. Since ZoneAlarm Security Suite 2010 uses Kaspersky’s virus-scanning engine, the anti-malware capabilities are very similar to those of Kaspersky’s product. However, the company has done a good job of integrating the virus-scanning technology into the suite, providing a near seamless experience from the program’s menus. Company Check Point Software Technologies Inc. Blocking malicious programs is easy, thanks to ZoneAlarm’s SmartDefense Advisor technology, which preconfigures settings for millions of known programs and sets a trust level for each. Unknown malware is handled by an “Auto-learn” mode, which starts by treating every unknown program as valid, allowing that program to be executed and then monitoring the activity of that program to see if it exhibits suspicious activity. Initially, all unknown applications have a rule assigned that allows continued network access. That leaves it up to ZoneAlarm to detect if the program is a problem, based upon behavior. The goal of Auto-learn mode is to limit confusing firewall pop-up messages, making security less intrusive — but with that comes the possibility of reduced security. If you turn off Auto-learn, then unknown programs are blocked until the user acknowledges that they are OK — which may be more irritating, but is also more secure. The product integrates with popular browsers and prevents malware by blocking dangerous sites. If the site is not blocked, the product allows the requested file to be downloaded. If it can’t guarantee the file is good, ZoneAlarm initiates a more intense scan that analyzes the file’s execution in a digital sandbox. The advanced scan starts after the download finishes and can take a few minutes. ZoneAlarm has done a fine job of rolling the separate security components together into a unified suite. I found it very easy to install. Dialog boxes were kept to a minimum, requiring very little user interaction — while that does simplify the installation, it would have been nice to be presented with a little more information, such as percentage complete and what part of the installation process was occurring. Scans proved to be very fast, and the application used a minimum of CPU cycles and resources, making it effective even on older systems with low-powered CPUs and on netbooks. In most cases, test scans only increased CPU utilization by 10per cent or less. However, more in-depth scans of executables, which execute the applications in a digital sandbox, spiked CPU usage as high as 90per cent for a few seconds. According to ZoneAlarm’s PR representative, the company has not released any information about the next version of its product or about its plans for an updated version of ZoneAlarm Security Suite 2010. All things considered, ZoneAlarm Security Suite 2010 covers the basics very well. Its integrated firewall proves to be an excellent security tool for power users who want to control and monitor all traffic in and out of a PC. I do have an issue with its Auto-learn mode — but as long as you ignore that feature, the firewall is very good. You should also expect performance hits during in-depth scans. Thanks to the incorporation of Kaspersky’s security tools, ZoneAlarm Security Suite 2010 will protect PCs from the common ills found on the Internet, which helps to round out the product and put it into the Internet Security Suite category. The product could be a top contender with the addition of free phone support, which is the norm among the vendors in this market, and a slimmed-down interface that better hides complexity from neophyte users.

What prospect does the pharmaceutical sector have for the healthcare and economic development of Nigeria? The Pharmaceutical sector has great potential for both diverse and interrelated sectors. As you are aware, we came up with the medicines’ security concept which says that unless a people exert sufficient control over how their medicines and healthcare commodities are produced, sustainable access to relevant, affordable, high quality products cannot be guaranteed in that setting. In line with this concept, our work is therefore positioned to expedite improved access to healthcare in the nation. Another critical area where we are focusing our efforts on is ensuring that our research and development initiatives are embedded in governments overarching socioeconomic objectives. As such we are re-engineering our projects to drive key government policies such as job creation, human capital development and revenue generation. The framework we are developing will therefore ensure that the Pharma Sector will not only improve access to health in Nigeria, it will increase other socioeconomic indices, such as employment generation, knowledge transfer, capacity building, attraction of foreign direct investment, and backward integration in ancillary industries. Is there evidence to support argument for prioritisation of the pharmaceutical sector in Nigeria? A number of things are happening in Africa which supports my argument for prioritisation of the sector. The continental pharma market is estimated to hit $50 billion per annum in a couple of years. Also, there is an ongoing paradigm shift, meaning that the policy architecture is gradually changing to favor local content development, both in terms of natural resources and human capital. Together, these two factors mean that great opportunities exist for entities with local presence. These, alongside other variables such as our considerable disease burden and high fertility rates, make the African pharma market one that cannot be ignored. It is therefore important to explore what this scenario portends for the country. Nigeria currently has over 120 manufacturers and 500 importers of pharmaceuticals and related products, who represent a significant proportion of all pharmaceutical companies in Africa. Nigerian pharma companies have also attained the highest standards in products as well as in processes. For instance, four out of 10 companies who have World Health Organisation GMP certification in sub-Saharan Africa are Nigerian companies. Additionally, Nigeria currently has one of the highest populations of professionals with technical capacity in various fields of pharmaceutical development. It is therefore clear that Nigeria is suitable positioned to emerge as the Pharma sector hub for Africa, and that means the potential to capture a significant proportion of the $50 billion a year market, which will in turn catalyse widespread development even in interrelated sectors. Up until now however, Nigeria has not been harnessing these resources that can position it to capture the continental market. With the framework we are propagating, our preliminary analysis indicates that the Nigerian pharma sector will, in addition to exponentially improving access to medicines, significantly contribute to the national Gross Domestic Product (GDP) as well as provide employment for over one million Nigerians. What are the challenges affecting prioritisation of the pharmaceutical sector in Nigeria? Previously, challenges affecting prioritisation of the pharma sector used to be political will, but the current administration has demonstrated through various policies and executive orders that the development of natural resources and local content remains the highest priority. Now what we need to address include getting various actors to understand the benefits of prioritising the sector and getting stakeholders with outlier interests to align with the national position. Doing this in a timely manner is however of utmost importance as Nigeria’s current vantage position is fast being eroded. Ghana recently passed the Executive Instrument Number 181 ultimately aimed at achieving the same goals, Ethiopia too has just launched a Pharmaceutical Sector Development Plan expected to replicate the success of Ethiopian Airlines’ success story in Aviation. It is important to note that the framework and strategy we are propagating has been successfully used in other settings. There is international evidence that similar approaches have been used in aggressively developing other pharma sectors, for instance in India, Thailand and Bangladesh. Also, Ethiopia and Rwanda have successfully used the same strategy to respectively develop their aviation and ICT sectors. What sort of socioeconomic benefits can be associated with the development of the Nigerian pharmaceutical sector? It is on record that the production of pharmaceuticals is associated with some of the highest value addition chains, in terms of backward integration and ancillary sector development. Empirical evidence exists that suggests that for every job created in the pharmaceutical manufacturing sector, between five to 10 corresponding jobs are created in the wider economy. This makes the pharma sector a powerful tool for stimulating employment generation. With prioritisation of the sector, the emergent framework would support mergers, acquisitions and partnerships that leverage on the current considerable infrastructure that exists in the sector. The opportunities thus created would generate sizeable interests from new investors as well as other global players in the industry, and consequently attract foreign direct investment to the economy. The framework and consequent recognition of Nigeria as the pharma manufacturing hub can even become a basis for anchoring emergent development initiatives for other sectors. With respect to backward integration, the prioritisation would further potentiate innovation and research initiatives already being spearheaded by the National Institute for Pharmaceutical Research and Development (NIPRD. For instance, activities on excipients like pharmaceutical grade starch, cellulose and alcohol, which can all be sourced locally from various natural resources, such as cassava and grains. The framework being developed will therefore enable a seamless nexus for these other local industries that provide such input to the Pharmaceutical Industry. Additionally, activities at the institute aimed at training local people to harness Nigeria’s abundant rich Natural Biodiversity, will be accelerated by the prioritisation of the sector. With the framework in place, targeted activities for the local production of Active Pharmaceutical Ingredients (APIs) from petrochemicals, as well as natural sources, can commence in earnest. The synergistic contribution of these various aspects of the framework will lead to a systematic expansion of the National economy. Expectedly, this will further boost employment generation, knowledge and technology transfer, revenue generation, export of Nigerian products and many other related socioeconomic indices. What is the role of NIPRD in formulating development strategy for the pharma sector in Nigeria? NIPRD is the only one of its kind in the region, and it is statutorily charged with the responsibility for research and development of drugs, vaccines, phytomedicines, commodities, and diagnostics aimed at improving sustainable access to safe, affordable and high quality healthcare. NIPRD also undertakes activities relating to capacity building, policymaking, data collation, drug distribution and the development of contextual partnerships that can expedite access to healthcare. To accelerate development in the sector, our reengineering strategy has commenced with the aggregation of the relevant data sets as well as exploration of the appropriate engagement models that can support the most efficient and effective transition in our setting. We have also begun working on the identification and classification of relevant stakeholders, as it is of critical importance to partner with parties that have similar goals and objectives, particularly with respect to increasing access to healthcare, as well as ensuring that the reforms are sustainable. Engagement within government as well as with external actors have now commenced as a prelude to the next phase. In our view that the pharmaceutical sector is suitably positioned for exponential growth which will no doubt translate to increased access to medicines for Nigerians, as well as stimulate development in various sectors within the country. The prioritisation and consequent reengineering of the sector is no doubt the most efficient and effective strategy for achieving this.

North Cork Junior A Hurling Championship Development The Cork Junior Hurling Championship had been contested on a countywide basis since 1895, however, an increase in the number of participating clubs resulted in a restructuring of the entire championship. The North Cork Board was established in 1925 with other divisions in other parts of the county created in the years that followed. Since then the individual junior championships were organised on a divisional basis with the division winners progressing to contest the county series of games. The first North Cork Junior Championship was played in 1925. In 2007 the championship was split in two with the top teams competing in the newly named North Cork Premier Junior Hurling Championship. Following this change the North Cork Junior A Hurling Championship was contested by the second tier teams. This system lasted until 2014 when the premier championship reverted to being called the North Cork JAHC. Team dominance Milford were the first team to enjoy multiple successes in the championship. They won four titles between 1925 and 1936, while also becoming the first club to retain the championship. Near neighbours and local rivals Ballyhea and Newtownshandrum enjoyed a dominant period from 1939 until 1968. The two clubs shared 14 championship titles during this period. Castletownroche also had their greatest period during this time, winning four titles between 1954 and 1960. Since the 1960s the championship was dominated by Kildorrery and Kilworth, who regularly won titles in each of the following decades. Kilworth claimed all of their 11 championships in a 45-year period between 1961 and 2006 to leave them in the top position on the all-time roll of honour. Their hegemony was closely challenged by Kildorrery who won eight of their nine championship titles between 1962 and 1988. At this time Fermoy emerged as a force, winning five championships between 1987 and 1999. Their county final defeat in 2000 brought the curtain down on their greatest era in the competition. The first decade of the 21st century was dominated by Charleville. After a 15-year hiatus they claimed their fifth ever title in 2001. Four more championships followed between 2002 and 2011. Format history For the first seventy years the championship was played as a single elimination tournament whereby once a team lost they were eliminated from the championship. Since the late 1990s the championship has seen the introduction of a “back door” system which provides each team with a minimum of two games before exiting the championship. Qualification for subsequent competitions The North Cork Championship winners and runners-up qualify for the subsequent Cork Junior Hurling Championship. Prior to 2017 only the winners were permitted to progress to the county championship. Overview The North Cork Junior Championship is effectively a knockout tournament with pairings drawn at random — there are no seeds. Each match is played as a single leg. If a match ends as a draw there is a period of extra time, however, if both sides are still level at the end of extra time a replay takes place and so on until a winner is found. The double Two teams have won the North Cork Junior A Hurling Championship and the North Cork Junior A Football Championship in a single year as part of a hurling-Gaelic football double. Fermoy became the first club to win the double when they achieved the feat in 1941. Clyda Rovers became only the second team to complete the double in 1989. Kildorrery, Mallow, Buttevant and Doneraile also hold the distinction of being dual North Cork Championship winning teams, however, these were not achieved in a single calendar year. Combined teams Glanworth-Harbour Rovers and Ballygiblin-Mitchelstown have also won North Cork titles in both codes.

Only four players scored for Texas Tech in its 69-47 loss to Kansas State on Wednesday, and three starters - Robert Lewandowski, Javarez Willis and DeShon Minnis - registered "goose eggs," as Tech coach Billy Gillispie described the scoreless outputs. Lewandowski's scoreless performance came three days after he tied a career high with 20 points in a loss to Iowa State. "You have to have more consistency," Gillispie said. "It's very difficult, because you need to get to the point where you know what you can get from guys. ... You just keep on keeping on and try to get better." Aside from Jaye Crockett's nine points, all of Tech's scoring on Wednesday came from true freshmen. The Red Raiders shot just 37.5 percent from the field (15 of 40), their seventh straight game under the 40-percent plateau. Making just his fourth start and second at home, Estacado product Kevin Wagner gave the local crowd reasons to cheer during Texas Tech's 69-47 loss to Kansas State. The freshman guard scored a career-high 16 points, including Tech's first five points that gave the Red Raiders a brief lead in the early minutes. Wagner also dished out four assists and when freshman Jordan Tolbert was asked what sparked his 18-point performance, he replied simply, "Kevin Wagner." "He played really well tonight and got me the ball," Tolbert said. Wagner's breakout performance wasn't without its shortcomings, though. He shot just 5 of 16 from the floor - including missed layups to begin the first and second halves - and turned the ball over five times. Still, his ability to get to the rim off the dribble was an element Tech has lacked in its offense this season. Like many of Tech's other players, he must improve his finishes around the basket. The team's inability to do so left Gillispie frustrated following Wednesday's game. "When you get a layup, you need to make it," Gillispie said. "Either that or get two foul shots. We just continue to miss layups." Texas Tech has been one of the nation's most turnover-prone teams this season, and those problems reached a fever pitch on Wednesday. The Red Raiders committed 14 of their season-high 25 turnovers in the first half, and Kansas State finished with 27 points off those miscues. Tech coach Billy Gillispie called the turnovers "demoralizing." "When you go down there and just give them the ball," Gillispie said ... "it just demoralizes you. I don't care how tough you are, it's demoralizing. You talk them through and coach them through it - say we have to be tough, we have to be resilient and move on to the next play - but when you just go down three times in a row when they can't score on you and bounce it off your foot or throw it to the defense, it's tough." "But we've got to work through it and we've got to get better."

Role of redox-sensitive catalytic interaction with ADAM10 in mutant-selective extracellular shedding of prion protein Misfolded glycosylphosphatidylinositol-anchored prion protein (PrP) is primarily degraded in lysosomes but is often rapidly removed from the cell surface before endocytosis in a preemptive manner. However, this mechanism is poorly understood. In this study, we discovered a disease-causing prion mutation (Q212P) that exceptionally promoted the extracellular release of PrP. Spatiotemporal analyses combined with genome editing identified the role of sheddase ADAM10 in Q212P shedding from the cell surface. ADAM10 was observed to catalytically interacts with Q212P but non-catalytically with wild-type PrP (wtPrP). This intrinsic difference in the interaction of ADAM10 between Q212P and wtPrP allowed Q212P to selectively access the sheddase activity of ADAM10 in a redox-sensitive manner. In addition, redox perturbation instigated the latent misfolding propensity of Q212P and disrupted the catalytic interaction between PrP and ADAM10, resulting in the accumulation of misfolded PrP on the cell surface. Upon recovery, active ADAM10 was able to reversibly release the surface Q212P. However, it might prove detrimental if unregulated resulting in unexpected proteotoxicity. This study provides a molecular basis of the mutant-selective shedding of PrP by demonstrating the catalytic interaction of ADAM10 with Q212P. Introduction Secretory and membrane proteins newly synthesized in the endoplasmic reticulum (ER) undergo folding, modifications, and a complex assembly during transit through the secretory pathway. This enables the proteins to acquire unique three-dimensional structures for efficient functioning. These processes require a sophisticated collaboration of various molecular chaperones and are prone to perturbation by mutations, translational errors, or different environmental stresses. In general, these perturbations restrict many newly synthesized proteins from folding properly and therefore, such proteins are efficiently removed by a timely activation of quality control (QC) mechanisms. However, some misfolded proteins may accumulate in secretory organelles or on the cell surface, leading to risks of proteotoxicity. ER-associated degradation (ERAD) is the most extensively studied and conceptualized QC mechanism in the stressed ER, in which thousands of misfolded substrates with varying biophysical parameters are recognized by adaptors in the ER lumen, delivered to the cytosolic side, extracted from the ER, and degraded in the proteasome. The first two steps are critical and therefore, tightly controlled in a substrate-specific manner in the ER lumen, while the next two occur in the cytosol and are shared by almost all substrates. Owing to this, at present, the major interest lies in elucidating the factors or mechanisms that discriminate potential ERAD substrates expected to be misfolded from a pool of newly synthesized proteins in the folding process. For example, misfolding of glycoproteins identified as the most extensively studied group of ERAD substrates is mediated by aberrant glycan modifications and is recognized by ER-resident lectins. However, not all misfolded proteins synthesized in the ER are degraded by ERAD. One such typical ERAD-resistant misfolding-prone protein is the prion protein (PrP) synthesized in a form covalently attached to the luminal side of the ER membrane by a glycosylphosphatidylinositol (GPI)-anchor. PrP is a mysterious protein for which the conformational conversion from its native cellular form (PrP C ) to its pathological isoform (PrP SC ) directly correlates with the pathogenesis of fatal neurodegenerative disorders. More than 30 variants in the human PrP have been identified as causes of prion diseases characterized with diverse heterogeneous phenotypes. Notably, these mutations often act during the conversion of PrP C and perturb its trafficking or processing, therefore, resulting in the accumulation of misfolded PrP in intracellular compartments during passage through the secretory pathway. Misfolded PrP accumulation may occurs due to inefficient clearance of conformationally problematic PrP mutants. This can be resolved by various QC mechanisms that are mechanistically different from ERAD and are rapidly activated in response to ER stress. For example, preemptive QC (pQC) aborts the production of difficultto-fold proteins, including PrP, at the step of protein translocation across the ER membrane. In one of our recent studies, we have revealed that pQC is utilized for mutant-selective topologic conversion (MSTC) of PrP, which selectively removes the membrane-anchored PrP isoform (ctmPrP) and alleviates PrP proteotoxicity. In addition, rapid ER stress-induced export (RESET) rapidly exports ERAD-resistant misfolded PrP from the stressed ER. This exported PrP forms a complex with ER-derived chaperones and cargo receptors, is escorted through the secretory pathway, and is transiently expressed on the cell surface before being subjected to lysosomal degradation. As such, both pQC and RESET are undoubtedly beneficial to reduce substrate burden and restore the folding capacity in stressed ER. More recently, we found that the endocytosis of PrP mutants defective in the disulfide bridge, a well-characterized RESET substrate, is inhibited at the cell surface as a result of the removal of the unpaired free-thiol group through the plasma membrane QC (pmQC) mechanism. pQC and pmQC act by reducing the burden of misfolded PrP entering the ER and lysosome, respectively. pQC is broadly utilized by the cells for many substrates in a signal sequence-dependent manner, whereas pmQC has a limited substrate range owing to its activation mediated in a conformation-dependent manner. A disintegrin and metalloproteinase 10 (ADAM10), a major sheddase along with its close relative ADAM17, has been identified as a potential modulator of the pmQC for PrP. ADAM10/17 are membrane-integrated proteins and ubiquitously expressed in mammalian cells. Thses proteins are able to function as molecular scissors and proteolytically cleave the extracellular regions or ectodomains of at least 40 integral membrane proteins playing important roles in cell adhesion, proliferation, differentiation, migration, immunity, and receptor-ligand signaling. PrP is the first GPI-anchored protein identified as a common substrate for ADAM10/17. Of these, ADAM10 has been reported to cleave the site in close proximity to the GPI-anchor linked to the C-terminus of surface PrP between the amino acid positions, Gly 228 and Arg 229, resulting in the release of anchorless PrP into the extracellular space. Nevertheless, the processes regulating the extracellular release of anchorless PrP mediated by ADAM10/17 remain poorly defined. In the present study, we found a naturally occurring pathogenic PrP mutant (Q212P) processed by pmQC. Spatiotemporal analysis of PrP expression, trafficking, and processing, combined with genome editing, identified ADAM10 as a pmQC regulator that triggered Q212P shedding from the cell surface. This study unraveled the molecular basis of this triage reaction that occurred at the cell surface in response to the environmental redox perturbations. This study also provided an insight into the ADAM10-mediated Q212P shedding as a peripheral QC to protect cells from the surface overload of misfolded PrP. Endo H, PNGase F, and all enzymes used for cloning were obtained from New England Biolabs. MG132, DTT, cycloheximide, thapsigargin and all chemicals used for the biochemical procedures in this study were purchased from Merck. Monensin and bafilomycin-A1 were purchased from MedChemExpress. Cell culture analyses Flp-In T-REx 293 cells were purchased from Invitrogen (R78007) and grown in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum and incubated in 5% CO 2 at 37 C. To establish CRISPR/Cas9 cell lines, Flp-In T-REx 293 cells were modified by the stable integration of the Cas9 gene (cloned in pcDNA3), which promotes genome editing. Ten Flp-In T-REx 293-Cas9 cell lines were established by the clonal isolation of G418-resistant cells by serial dilution in 96 well plates, and their Cas9 activities were verified by T7E1 assay and immunoblotting. Isogenic cell lines with an inducible expression of stably integrated wild-type or mutant PrPs were generated according to the manufacturer's directions (Invitrogen) following transfection of plasmid constructs with Lipofectamine 2000 (Invitrogen). In this system, PrP expression was controlled by the CMV promoter activated by doxycycline (Dox) (10 ng/ml) for 16 h unless otherwise indicated. To establish an ADAM10-deficient cell line (ADAM10), the Flp-In T-REx 293-Cas9 cell line was transfected with a mixture of three ADAM10 and ADAM17 gRNA constructs as described (see "Molecular biology" section) for 24 h and incubated in the presence of puromycin (2 g/mL) for an additional 48 h. Viable cells were maintained in culture media containing puromycin (0.5 g/mL). Subcellular localizations of PrPs were analyzed by directly visualizing the fluorescence in cells cotransfected with PrP fused with RFP and GFP as previously described, and images were obtained using a confocal microscope (Zeiss LSM880 with Airyscan; Carl Zeiss Microimaging) using the manufacturer's image acquisition software, ZEN 2.3 (black edition). Colony forming assays were performed using a previously published procedure with minor modifications. Briefly, 100 cells per well were plated on 6-well dishes and cultured in DMEM supplemented with Dox for three weeks. Viable cell colonies were fixed and counterstained with 0.5% crystal violet in 6% glutaraldehyde, and visualized via GelCount™ (Oxford Optronix; Abington, UK) using the manufacturer's image acquisition software. Molecular biology The PrP coding region (GeneBank accession number: M13899) was PCR-amplified from the first-strand cDNA synthesized from HeLa cells and cloned into the HindIII and XhoI sites of the pcDNA5-FRT/TO vector (Invitrogen). All mutant constructs used in this study were engineered by conventional site-directed mutagenesis using Phusion high-fidelity DNA polymerase (New England BioLabs). Fluorescent protein (FP) fusion constructs were created by the insertion of GFP or RFP genes into the unique Bsu36I site within the N-terminal coding region of wtPrP and mutant PrP. ADAM10 constructs were created by the insertion of human cDNA encoding ADAM10 (GeneBank accession number: AF009615) into HindIII and XhoI sites of homemade pcDNA5-FRT/TO-HA vector. A mutant ADAM10 (AXXA) construct was constructed in the same manner as the PrP mutants. Biochemistry Pulse-chase experiments were performed to determine PrP synthesis and turnover rates. PrP expression was induced by Dox (10 ng/ml) for 16 h before pulse-labeling. Cells were starved with serum-free and methionine/cysteine-free media for 30 min and pulse-labeled with a trans-labeling mixture (-methionine/cysteine; PerkinElmer) for an additional 30 min. The spent medium was replaced with fresh growth medium, and the pulse-labeled cells were solubilized in buffer K (1% SDS, 100 mM Tris-HCl, pH 7.5) at the indicated time points and diluted to 10-folds with IPT buffer (1% Triton X-100, 50 mM HEPES, pH 7.5, 150 mM NaCl). PrP was captured by PrP-specific 3F4 antibody or anti-RFP for 90 min and recovered from cell lysates by protein G-conjugated magnetic beads (GE Healthcare Life Sciences) for an additional 90 min. The beads were washed five times with 1 mL of IPT buffer to remove nonspecific binding and resuspended in 30 l of 1.5X SDS-PAGE sample buffer. Lysosome-restricted free RFP generation was assessed by monitoring the level of free RFP produced from newly synthesized intact PrP-RFP during the chase as previously described. For immunoprecipitation (IP) of PrP-RFP in the extracellular space, cell culture media was collected, passed through the 0.45 m syringe filter, and denatured by adding 0.1 volume of buffer K. The CM was incubated with an RFP-specific antibody and protein A-agarose beads for 90 min. The beads were washed five times with 1 ml of IPT buffer and resuspended in 30 l of 1.5X SDS-PAGE sample buffer. In some experiments (Fig. 6), the CM was further analyzed by sequential centrifugation. The CM containing PrP-RFP was subjected to centrifugation at 300g for 10 min. The supernatant was collected and further separated in a stepwise manner at 2000g to 200,000g for 20 min up to 1 h as described in Fig. 6C. The PrP-RFP was enriched in the supernatant or pellet from each centrifugation and detected by immunoblotting with an RFP-specific antibody. For a stable isolation of PrP complexes expressed on the cell surface, surface-restricted labeling using GFP-Nb was performed as previously described with minor modification. Briefly, PrPs-GFP were induced by the treatment with Dox (10 ng/ml) for 24 h, and these complexes were covalently linked with their interacting partners using 2 mM of DSP (ThermoFisher, 22,585) for 30 min at room temperature (RT). Following quenching with 20 mM of Tris (pH 7.5) for 15 min at RT, surface PrPs-GFP were selectively labeled with 100 nM FLAG-conjugated GFP-Nb prepared in 1X PBS supplemented with 10% FBS for 30 min at 4 C. The labeled cells were solubilized in lysis buffer (1% CHAPS, 50 mM HEPES, 150 mM NaCl, 2 mM MgCl 2 ) for 10 min on ice and centrifuged at 16,000g for 10 min to remove cell debris. The supernatant was further incubated with anti-FLAG M2 magnetic beads (Merck M8823) for 3 h at 4 C. The beads were washed six times with lysis buffer, and the PrP-interacting partners crosslinked with surface PrP-GFP were eluted by the incubation with 30 L of lysis buffer containing FLAG peptide (1 mg/mL) for 30 min at RT. In all experiments involving IP, 10 L of eluents were separated on gels and analyzed by autoradiography or immunoblotting with appropriate antibodies. Miscellaneous Comparative analyses of biochemical properties including glycosidase sensitivity, detergent solubility, trypsin sensitivity, and sucrose gradient centrifugation were performed as previously described. In protein analysis experiments, the cells were fully solubilized in buffer K, boiled, and separated by SDS-PAGE using 10 or 12% Tris-tricine gels. The experimental conditions for each experiment have been described in individual figure legends. Quantitation and statistical analysis Average band intensities of immunoblotting were measured directly by Image Studio (LI-COR, Inc.), and those of 35 S incorporation into the newly synthesized proteins were inverted to black-and-white for clarity and measured by Image J (NIH) software. The calculations and protein expressions have been described in individual figure legends. The data has been presented as the mean ± standard deviation (S.D.) of more than three independent replicates. n represents the number of independent experiments. Statistical differences were analyzed using Student's t-test, and their significance was expressed with p-values as follows; *p < 0.05, **p < 0.005, ***p < 0.001. Q212P mutation promotes PrP trafficking and processing This study was motivated by the discovery of a disease-causing PrP mutant (Q212P) showing a unique pattern of PrP subpopulations. Most notably, in contrast to wtPrP synthesized in four different forms corresponding to non-, mono-, di-, and fully glycosylated forms, Q212P was synthesized in two major forms, i.e., di-and fully-glycosylated forms (Fig. 1A). Of these two forms, the fully-glycosylated form modified by complex glycan while passing through post-ER compartments during the secretory pathway (i.e., as it is sensitive to PNGase F but resistant to Endo H) was selectively augmented by ~ 1.6-fold as a result of Q212P mutation (Fig. 1A, see also Fig. S1A). This pattern is not generally observed in pathogenic PrP mutants, as shown by the H187R mutant, which did not synthesize the fully glycosylated form (Fig. S1A). Nevertheless, the general pattern in subcellular localization of PrP was unchanged by the Q212P mutation, as visualized by the complete overlap of Q212P-RFP and wtPrP-GFP at the cell surface and intracellular compartments (Fig. S1B). In contrast, the surface localization of PrP was selectively inhibited by Q212P mutation through repressed de novo protein synthesis using cycloheximide (CHX), suggesting a possible role of Q212P mutation in promoting the clearance of surface PrP (Fig. 1B). To explore this idea, we investigated the impact of the Q212P mutation on PrP synthesis and turnover using pulse-chase analysis. In this experiment, inducible expression of stably integrated untagged PrP allowed us to efficiently discriminate the small differences in sizes of folding intermediates transiently synthesized in transit through the secretory compartments. Consistent with the result of immunoblotting (Fig. 1A), wtPrP was newly synthesized in non-, mono-, and diglycosylated forms in pulse-labeled cells, whereas the Q212P mutant was synthesized predominantly in a di-glycosylated form without detectable amounts of the other forms (Fig. 1C). During the chase, the diglycosylated forms of both wtPrP and Q212P were trimmed with glucosidases in the ER within 30 min and further modified with complex glycans in the Golgi apparatus within 1 h. In contrast, monoglycosylated wtPrP remained unmodified further in the ER until 4 h after the chase. Despite the similarity in the rate of post-ER trafficking, the fully glycosylated PrP was produced at a higher level in Q212P cells than in wtPrP cells. This could be attributed to the fact that the post-ER trafficking was allowed only for the di-glycosylated form initially synthesized in pulse-labeled cells, noting that a higher level of diglycosylated PrP was newly synthesized in pulse-labeled Q212P cells (Fig. S1C). Thus, an accelerated PrP maturation mediated by the Q212P mutation might be a plausible explanation for the observed increase in fully glycosylated Q212P. Q212P mutation triggers the extracellular release of PrP In our pulse-chase analyses, we noticed that fully glycosylated PrP was decreased selectively in Q212P cells by ~30% between 2 h and 4 h after the chase ( Fig. 1C and D). Given the fact that lysosomes are the primary sites for the clearance and recycling of PrP, the logical explanation for the decrease in surface Q212P may be attributed to inhibitory endocytosis. To investigate this idea, we employed a well-validated method that has been previously used to determine the differential rate in lysosomal deliveries of wtPrP and mutant PrP inserted by RFP resistant to lysosomal acidic proteases. We reasoned that, by monitoring the level of free RFP in cells expressing RFP-inserted Q212P, we could verify the impact of the Q212P mutation on proteolytic endocytosis of PrP ( Fig. 2A). A combination of this assay with pulse-chase experiments revealed that RFP insertion did not perturb the general pattern of PrP synthesis and turnover; the newly synthesized wtPrP-RFP was fully glycosylated within 1 h after the chase and then endocytosed and processed into free RFP (Fig. S2A). The fully glycosylated PrP was expressed on the cell surface within 1 h after the chase, as deduced through its complete digestion by exogenously added trypsin (Fig. 2B). In contrast, free RFP was protected by trypsin and increased progressively up to 4 h after the chase; however, its generation was suppressed by an inhibitor of lysosomal acidic proteases, bafilomycin A1 (BAF-A1) (Fig. 2C, see also Fig. S2B). Accelerated proteolytic endocytosis did not appear to be the sole reason for the reduction in surface Q212P owing to the unchanged level and rate of free RFP generation post its mutation (Fig, 2B, see also Fig. S2A). This might be a reasonable explanation for the increase in accumulated Q212P accumulated at the cell surface (Fig. 1A). Therefore, we aimed to identify an additional mechanism by which Q212P is selectively processed at the cell surface. We hypothesized that extracellular PrP release might also reduce the surface PrP levels. To test this hypothesis, we pulse-labeled cells, collected the conditioned media (CM), and examined whether the newly synthesized PrP was recovered from the CM by an RFP-specific antibody. As expected, newly synthesized PrP-RFP, but not free RFP, was selectively recovered from the CM of Q212P cells. Extracellular Q212P proved to be a fully mature form synthesized through the conventional secretory pathway, as revealed by its similarity in gel mobility to the Endo H-resistant surface PrP (of note, Endo H selectively cleaves simple modifications of N-linked glycans processed by ER enzymes) (Fig. 2D) and the inhibition of its secretion by monensin (of note, monensin prevents protein secretion from the medial to trans cisternae of the Golgi complex) (Fig. 2E). Thus, Q212P mutation might be able to catalyzes the extracellular release of surface PrP without affecting endocytosis. ADAM10 catalyzes Q212P shedding at the cell surface We aimed to identify the factor responsible for promoting the extracellular release of surface Q212P and first hypothesized that sheddase ADAM10, a surface metalloproteinase that proteolytically cleaves the ectodomain of surface PrP was a promising candidate. This hypothesis was supported by two notable preliminary results. First, surface PrP was shed by N-ethylmaleimide (NEM) (Fig. S3A), a thiol-modifying reagent that is reported to activate sheddase ADAM10. Second, the extracellular Q212P level was decreased by GI254023X (Fig. S3B), a potent inhibitor of ADAM10. To obtain more direct evidence, we established an ADAM10disrupted (ADAM10) cell line using the CRISPR/Cas9-mediated gene editing. Defective gene expression and protein elimination of ADAM10 were confirmed by RT-PCR and immunoblotting, respectively (Fig. 3A). This cell line was further engineered with inducible expression of stably integrated Q212P-RFP. We monitored the newly synthesized Q212P-RFP recovered in the cell lysates or the CM to determine whether ADAM10 was required for the steady-state shedding of Q212P. Comparative analysis of extracellular Q212P-RFP newly synthesized from pulse-labeled wild-type (WT) and ADAM10 cells showed a clear inhibition of Q212P shedding in ADAM10 cells (Fig. 3B, left panel, see also Fig. 4C). This inhibitory Q212P shedding conferred an increase in the level of surface Q212P but was not reversed by NEM (Fig. 3B, right panel, see also Fig. 4C). In contrast, the general pattern of free RFP generation was changed neither by ADAM10 alone nor in combination with NEM (Fig. 3B), consistently suggesting that ADAM10 was not functionally involved in Q212P endocytosis (Fig. 2B, see also Fig. S2A). In this context, NEM-induced PrP shedding differed from the steadystate Q212P shedding in two notable aspects. First, it was not specific for Q212P, and second, it inhibits PrP endocytosis as judged by the decreased level of free RFP during the chase. Moreover, the sheddase activity of ADAM10 for Q212P appeared to be exerted at the cell surface. This was revealed by the inducible reexpressions of wt and mutant (i.e., CXXC-to-AXXA substitution) ADAM10 in ADAM10 cells. Q212P shedding was restored by the reexpression of wtADAM10, but not by that of the mutant ADAM10 (Fig. 3C) previously shown not to be expressed on the cell surface by being converted into the inactive conformation. An additional result in line with our biochemical analyses was also observed in cells chased with CHX and verified by direct visualization of Q212P-RFP restored on the cell surface of ADAM10 cells (Fig. 3D). Collectively, these observations suggested that the Q212P mutation promoted the Fig. 2. Analysis of Q212P release into the extracellular space (A) The lysosome-restricted free RFP generation. The expected metabolism and processing of PrP-RFP were indicated by green arrows; the differential rate in lysosome deliveries were assessed by monitoring the level of free RFP in cells expressing RFP-inserted PrP. Reagents used in this study for determining the localization of PrPs were shown in blue, and the extracellular release of surface Q212P was indicated by the red arrow. (B) The effect of Q212P mutation on the secretory trafficking of PrP was assessed by the surfacerestricted protease digestion. Cells expressing wtPrP-RFP and Q212P-RFP were pulse-labeled for 30 min and followed by the chase as described in the protocol (upper panel). At the indicated time points, intact radioactive cells were incubated for 10 min in the absence (-Trypsin) or presence (+Trypsin) of exogenously added trypsin (0.25%). Fully solubilized cells were diluted 10-fold with IPT buffer and subjected to immunoprecipitation with an RFP-specific antibody (1:500). Note, fRFP: free RFP resistant to lysosomal acidic protease. Surface Q212P catalytically interacts with ADAM10 Given the catalytic activity of ADAM10 on Q212P shedding, we inferred that ADAM10 discriminated Q212P mutation through direct interaction. To this end, we created stable cell lines expressing PrPs inserted by GFP (PrPs-GFP). In the preliminary experiment, we subjected these cell lines to a conventional co-IP and examined the interaction of PrPs with endogenous ADAM10. The cells used in this study synthesized two different endogenous ADAM10 corresponding to the intracellular and surface forms whose pro-domains were uncleaved and cleaved by proprotein convertases, furin and PC7, respectively. Of these two forms, the intracellular form (i.e., the precursor form) was observed to interact selectively with Q212P, suggesting that misfolded Q212P may be escorted by ADAM10 while passing through the secretory pathway. In contrast, the surface form appeared to interact with neither wtPrP nor Q212P (Fig. S4A). This may be attributed to the Q212P shedding is mediated by a rapid enzymatic reaction resulting from its transient interaction with ADAM10. This technical problem could be solved by stabilization of the interaction using DSP, a reversible crosslinker, followed by surfacerestriction labeling with an FLAG-conjugated GFP-Nb (FLAG-Nb). Using this method, the surface protein complexes containing PrPs-GFP were labeled selectively with Nb, recovered successfully by anti-FLAG antibody (Fig. S4B), and analyzed by immunoblotting with ADAM10specific antibody (Fig. 4A). Given that ADAM10-mediated PrP shedding occurred at the cell surface (Fig. 3C), surface ADAM10 (i.e., active form lacking pro-domain) was crosslinked with both wtPrP and Q212P and successfully recovered by Nb. However, the amount of ADAM10 recovered with Q212P was noticeably less than that with wtPrP. This could be attributed to the rapid extracellular release of cleaved Q212P (Fig. 4B). In contrast, ADAM17 (Fig. S4C), the structural and functional homolog of ADAM10, was observed not to interact with Q212P or wtPrP (Fig. 4B) and did not appear to be involved in Q212P shedding (Fig. 4C, see also Fig. S4D), indicating that PrP is not the substrate of ADAM17. This was shown by pulse-chase experiments in ADAM17 cells in the presence and absence of NEM (Fig. 4C, see also Fig. S4D). Given the model of ADAM10-mediated Notch proteolysis to be discussed later, we hypothesized that ADAM10 activity could be catalyzed by the interaction with Q212P, in a manner similar to NEM, but not by wtPrP. Q212P shedding is redox-sensitive Early nuclear magnetic resonance (NMR) structures and X-ray absorption measurements highlighted a unique feature in the secondary structure of Q212P distinctive to that of wtPrP. They revealed that its C-terminal region was readily convertible from its native to a pathogenic form. However, we failed to identify any noticeable difference in the two typical determinants indicating PrP proteotoxicity, detergent solubility, and protease sensitivity between wtPrP and Q212P under normal conditions and in ADAM10 (Fig. S5A). Instead, unlike wtPrP, newly synthesized Q212P appeared to be sensitive to Fig. 3. Analysis of ADAM10-mediated Q212P shedding (A) ADAM10-deficient cells expressing Q212P-RFP were established using CRISPR/Cas9-mediated genome editing (ADAM10) and verified by RT-PCR (left panel) and immunoblotting with ADAM10specific antibody (right panel). Note, X: non-specific band (B) The effect of ADAM10 on Q212P shedding was analyzed by pulse-chase experiments. Wild type (WT) and ADAM10 cells expressing Q212P-RFP were pulse-labeled for 30 min and allowed to recover for 1.5 h ("C1"), followed by the chase for an additional 4.5 h ("C2") in the absence or presence of NEM as indicated. The fully solubilized cells and CM were subjected to immunoprecipitation with an RFPspecific antibody. Plotted are the changes of intact Q212P-RFP (left panel) and free RFPs (middle panel) in cells and CMs (right panel) by NEM (5 mM) treatment (n = 3), and significance was considered with p-values as described in "Materials and Methods". Note, "N⋅S." means "Not Significant" (C) The importance of surface expression of ADAM10 in Q212P shedding was analyzed. ADAM10 cells expressing Q212P-RFP were transfected with constructs expressing wild-type or mutant (AXXA) ADAM10 fused with HA epitope and subjected to pulse-chase experiments as described in the upper panel. At the indicated time points, the cells and CM were harvested and immunoprecipitated with an RFPspecific antibody (1:500). The expression of transfected ADAM10 was detected in the fully solubilized cells by immunoblotting with an anti-HA antibody (1:5000). The plots represent the relative amount of Q212P-RFP in the CM of cells expressing wild-type versus mutant ADAM10 harvested at 5 h after the chase was plotted (n = 3; p = 0.017) (D) Surface expression of wtPrP-RFP and Q212P-RFP was visualized by direct RFP fluorescence in wildtype and ADAM10 cells following treatment with cycloheximide (100 g/mL) for 1 h. Scale bar: 50 m. environmental redox perturbations and was detected with additional high-molecular-weight smears by DTT (Fig. 5A) previously shown to inhibit intramolecular disulfide bonds within PrP. These bands were found to be SDS-resistant and confirmed to undergo PNGase F-resistant glycan modifications at least in part during passage through the Golgi apparatus (Fig. S5B). Further analysis of the native size on sucrose gradient suggesting the oligomerization propensity of Q212P under redox perturbations suggested the possibility that these bands were early intermediates of oligomeric Q212P (hereafter, referred as "Q212P*") (Fig. 5B). These results collectively suggested that Q212P had an intrinsically vulnerable conformation to the environmental redox perturbations. We further investigated the effect of DTT on Q212P shedding using the CM analysis. As expected, the newly synthesized Q212P* remained unreleased from the surface of the DTT-treated cells. Upon DTT withdrawal, the Q212P* was reversibly shed from the surface and increased in the CM with a proportional decrease in the surface level (Fig. 5C). Accelerated endocytosis and ER stress were not involved in these reactions, as revealed by the unchanged free-RFP levels and failure to generate Q212P* by an ER stress inducer thapsigargin (Tg), respectively (Fig. S5C). In a similar manner, we analyzed Q212P in the ADAM10 cells. Consistent with the observation in the WT cells (Fig. 5C), Q212P* was also observed in ADAM10 cells treated with DTT and was more prominent than in WT cells. However, unlike WT cells, the inhibitory shedding of the Q212P* was hardly restored in ADAM10 cells, even by DTT withdrawal (Fig. 5D). This finding was a plausible explanation for the increase in Q212P* on the surface of ADAM10 cells. The redoxsensitive inhibition of Q212P shedding was, therefore, reversible as long as ADAM10 was active (Fig. 5E). Inhibitory Q212P shedding promotes cell proliferation Given that oligomer-prone PrP exacerbated neurotoxicity and neuronal degeneration, persistent Q212P shedding by uncontrolled ADAM10 activity could eventually be detrimental unless extracellular Q212P was efficiently eliminated by a clearance mechanism. In contrast, inhibitory Q212P shedding might be more favorable to cells as misfolded surface PrP can be endocytosed eventually into lysosomes for degradation. This was validated by the ability of ADAM10 to promote colony formation in single cells expressing Q212P (Fig. 6A). Comparable results were also observed in long-term cultures of mixed cells expressing fluorogenic PrPs and verified by the predominant visualization of ADAM10 cells expressing Q212P-RFP (Fig. S6). These observations suggested a potential anti-proliferative effect of extracellular Q212P, with our results consistently demonstrating the efficient endocytosis of Q212P despite ADAM10 (Fig. 6B). This was hinted by the sequential centrifugation of the CM (Fig. 6C) showing that the extracellular Q212P was mostly insoluble and precipitated by ultracentrifugation at 100,000g (P100) and 200,000g (P200) (Fig. 6D). Intriguingly, ADAM10 was also detected in the CM. Extracellular ADAM10 was apparently released from the cell surface but appeared to be independent of Q212P shedding for the following reasons. First, this reaction was not Q212P-selective, as evident through the detection of ADAM10 in the CM of wtPrP cells (Fig. 6D-left and Fig. 6D-middle). Second, ADAM10 was recovered predominantly in S200 rather than P200, indicating that, unlike Q212P, it was mostly present in a soluble form (Fig. 6D-middle and Fig. 6D-right). Last, extracellular ADAM10 was the same in size as the surface form (Fig. 6D-left). In addition, we detected Hsp 90, a major cargo contained in the extracellular vesicles (EVs) such as exosomes, in the ADAM10-enriched S200 of both wtPrP and Q212P cells (Fig. 6D-right). This result was in agreement with a previous report suggesting that ADAM10 is incorporated into EVs and spontaneously secreted. Besides Q212P and ADAM10, two small fragments were additionally detected in S200 by an RFP-specific antibody (Fig. 6D-right). Considering that RFP was inserted into the N-terminal region of PrP, these WT, ADAM10, and ADAM17 cells expressing wtPrP-RFP and Q212P-RFP were pulse-labeled for 30 min and allowed to recover for 1.5 h, followed by the chase for an additional 4.5 h in the presence of NEM as in Fig. 3B. The fully solubilized cells and CM were subjected to immunoprecipitation with an RFPspecific antibody (1:500). (D) The catalytic interaction of ADAM10 with Q212P at the cell surface is described. fragments were most likely those of the N-terminal PrP (i.e., N1) proteolytically processed by various proteases and liberated from the cell surface in soluble form. Even though one such typical protease responsible for this cleavage (i.e., -cleavage) is ADAM10, N1 fragments were still produced in ADAM10 cells (Fig. 6E). Therefore, ADAM10 was more actively involved in ectodomain shedding near the site of GPI-anchor addition in the C-terminus than -cleavage within the central hydrophobic region of PrP. Discussion In this study, we illustrated mutant-selective extracellular release of surface PrP by showing spatiotemporal differences in the metabolism and processing of PrP attributed to the catalytic activity of sheddase ADAM10. The new findings are twofold. One is the discovery of a disease-causing human mutation (Q212P) that promoted ectodomain shedding of surface PrP. The other is the molecular basis for the susceptibility of ADAM10-mediated Q212P shedding to environmental redox perturbations. Our intensive biochemical analysis combined with mutational analyses provided clues to several unresolved issues related to the peripheral pmQC aimed to remove ERAD-resistant misfolded GPIanchored PrP (Fig. 7). Steps susceptible to Q212P mutation during the PrP metabolism pmQC is a regulated process that limits the endocytic flux of misfolded PrP into lysosomes from the cell surface for degradation. This endocytosis is beneficial to cells as it reduces the burden of lysosomes owing to the misfolded PrP, therefore ensuring the safety of the endocytic pathway. However, the physiologically relevance of this regulation was questionable, as all studies were focused on artificial mutants lacking one or both cysteine residues within PrP. Therefore, we aimed to identify pmQC clients from ~30 disease-causing PrP mutants that are reported to metabolize and process differently than wtPrP. We, for the first time, reported the Q212P mutation that selectively increases fully glycosylated PrP. There are various possible reasons for the relative increase in fully glycosylated Q212P (Fig. 1A). First, glycosylation-defective subpopulations might be degraded by ERAD. However, this is least likely because PrP is synthesized with covalent attachment to the ER membrane via a GPI-anchor and poorly degraded by ERAD. Second, the Q212 mutation might inhibit the endocytosis of misfolded PrP from the cell surface into the lysosome for degradation. This did not appear to be the principal reason because the Q212P mutation did not interfere with lysosome-restricted free RFP generation in our analyses (Fig. 2B). Third, the Q212P mutation might enhance the rate of PrP transport to ensure the safety of compartments involved in the secretory Fig. 5. Analysis of redox-sensitive Q212P shedding (A) Q212P* synthesis induced by DTT was analyzed by pulse-chase experiments. Cells expressing wtPrP-RFP and Q212P-RFP were pulse-labeled followed by the chase for 1 h in the absence or presence of DTT as indicated. The fully solubilized cells were subjected to immunoprecipitation with an RFP-specific antibody. Equal loading was confirmed by the detection of total proteins newly synthesized. Note, *: Q212P* (B) A higher oligomerization propensity of Q212P* was analyzed by the density gradient centrifugation. Cells expressing Q212P-RFP were treated with DTT (10 mM) for 1 h, solubilized in IP buffer containing 1% CHAPS, and separated on the sucrose gradient (5-25%) for 3 h in TLS-55 rotor at 50,000 rpm. Each fraction from the top to the bottom of the sucrose gradient was blotted with an RFP-specific antibody (1:5000). Note, T: unfractionated protein samples (0.1 vol) (C) Q212P* shedding was analyzed by pulse-chase experiments. Cells expressing Q212P-RFP were pulse-labeled for 30 min, chased in the absence ("-DTT") or presence of DTT (10 mM) ("+DTT") for 1 h, and recovered for the indicated time after DTT withdrawal (upper panel). Cells were fully solubilized and subjected to immunoprecipitation with an RFP-specific antibody (D) The effect of ADAM10 on Q212P* shedding was determined in ADAM10 cells in the same method as described in (C). The plots represent the changes of surface Q212P-RFP (left panel) and free RFPs in cells (middle panel) and CMs (right panel) by ADAM10 depletion (n = 3). The significance was analyzed through p-values calculated using the student's-t test as described in "Materials and Methods". (E) Redox-sensitive Q212P shedding is described. pathway. This was validated by the accelerated complex glycan modifications of Q212P compared to that of wtPrP in the pulse-chase analyses (Fig. 1C). Last, the Q212P mutation possibly inhibited the release of surface PrP into the extracellular space. Unexpectedly, the extracellular release of newly synthesized PrP was rather triggered by the Q212P mutation but was insufficient to completely remove the misfolded PrP from the cell surface (Fig. 2). These explanations allowed us to conclude that the rate of secretory trafficking (i.e., RESET) and extracellular release (i.e., pmQC) determine the surface level of pathogenic Q212P, as further discussed in Supplementary Note. Effect of modulation of ADAM10 activity on Q212P shedding Given the fact that the Q212P mutation promotes PrP shedding, we inferred that ADAM10 would interact with Q212P at the cell surface with a higher affinity than with wtPrP. However, their transient interaction on the cell surface posed a major challenge to validating this idea. Our attempt to stabilize the cell-surface interaction was able to successfully detect ADAM10 bound to PrP, but this interaction was apparently attenuated by the Q212P mutation. A logical situation supporting this observation would be a rapid release of surface Q212P from the cell surface immediately after the transient catalytic interaction with Fig. 6. Analysis of the proliferation-promoting effect of inhibitory Q212P shedding (A) The effect of inhibitory Q212P shedding on cell proliferation was determined by the colony-forming assay. Wild-type and ADAM10-deficient (ADAM10) cells expressing Q212P-RFP were cultured for 3 weeks in the presence of Dox, and allowed to form colonies, that were visualized by staining with crystal violet (left panel). The relative number of large colonies (>2 mm diameter) out of total colonies from three independent experiments (three times in an independent experiment) are plotted as a percentage, and significance was considered with p-values (left panel). (B) The potential proliferation-promoting effect of inhibitory Q212P shedding is illustrated (C) The protocol shows the sequential centrifugation of CMs containing wtPrP-RFP and Q212P-RFP. (D) The solubility of extracellular Q212P was examined by sequential centrifugation. Cells expressing wtPrP-RFP and Q212P-RFP were induced by Dox treatment for 24 h and subjected to centrifugal fractionation as described in (C). PrPs-RFP concentrated in indicated fractions were detected by immunoblotting with an RFP-specific antibody. The plots represent the changes of surface and extracellular Q212P-RFP in indicated fractions by ADAM10 depletion (n = 3), and significance was considered with p-values as described in "Materials and Methods". (E) CMs of wild-type and ADAM10 cells expressing Q212P-RFP were centrifuged at 200,000g for 1 h. The supernatant (S200) and pellet (P200) were subjected to immunoblotting with an RFP-specific antibody. The plots represent the changes of Q212P and N1 fragments in CMs by ADAM10 depletion (n = 3). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.) Fig. 7. A working model depicting the role of ADAM10 in promoting Q212P shedding On the cell surface, the PrP can interact with the cysteine-rich region of ADAM10, either catalytically or non-catalytically. Q212P mutation provides catalytic interaction of PrP with ADAM10, switching the conformation of the cysteine-rich region, allowing PrP to access the ADAM10 active site, and inducing its extracellular shedding. Under redox perturbation, inhibitory Q212P shedding induces the surface accumulation of oligomeric Q212P but is restored, albeit somewhat slowly, upon redox recovery as long as ADAM10 is activated. Nevertheless, cells prefer ADAM10-independent endo-lysosomal degradation to eliminate surface Q212P because it is more efficient than ADAM10-dependent extracellular shedding. This strategy renders cells more capable of fully digesting misfolded surface Q212P that may exert unexpected protein toxicity when released into the extracellular space. ADAM10. This may be key to the molecular basis of mutant-selective PrP shedding, as supported by the facts suggested in previous studies as follows. First, it is possible that the conformation of ADAM10 is switched from the resting state to the active state mediated by its interaction with Q212P. This idea was suggested by the model of ADAM10-mediated Notch proteolysis derived from the X-ray crystal structure of the mature ADAM10 ectodomain. Most notably, the ADAM10 catalytic site adopts a closed and autoinhibited conformation in the resting state. At this stage, the metalloproteinase domain is partially closed by an intramolecular cysteine-rich domain, limiting the access to the active site. In contrast, upon ligand binding, the activated Notch receptor opens and directly contacts the ADAM10 catalytic site to promote cleavage. Substrate selectivity is, therefore, not only a cause of a typical interaction of the substrate and ADAM10 but also the conformation of the substrate-binding pocket of ADAM10. Likewise, the Q212P mutation may indicate an open conformation of this pocket, which is compatible with a potential mechanism of the mutant-selective PrP shedding of ADAM10. Second, disulfide exchange switches the conformation of the cysteine-rich region and modulates substrate access to the ADAM10 active site, governing the catalytic activity and substrate selectivity of ADAM10. This has been previously proven by the conformation-specific antibody 8C7, which binds to a conformationally active cysteine-rich domain and inhibits dynamic shuffling of the disulfide linkage with a thioredoxin CXXC motif. As predicted from the structure, inhibitory disulfide shuffling keeps the ADAM10 active site open and enhances its catalytic activity on the substrate, for example, inhibiting Notch activity. In our study, for PrP, comparable results were observed by the exposure to the thiol-modifying reagent NEM, which inhibited dynamic rearrangement of disulfide exchanges; however, in contrast to the steady state, wtPrP was also shed at a similar efficiency to Q212P (Fig. S3A). Although the structural basis for the opening of the ADAM10 active site by the Q212P mutation remains unsolved, our biochemical analyses allowed us to speculate that, similar to the actions of 8C7 and NEM, the Q212P mutation inhibited disulfide exchange in the cysteine-rich region, relieved ADAM10 autoinhibition, and catalyzed ADAM10 activity for PrP shedding. Last, Q212P shedding is also likely to be also regulated by tetraspanins (Tspans) that form microdomains where ADAM10 is predominantly localized. There are 33 reported human Tspans (i.e., Tspan1-Tspan33) that interact with membrane proteins including cell adhesion, signaling molecules, and proteolytic sheddase such as ADAM10 to regulate their intracellular trafficking, lateral mobility, and cell surface clustering. Unlike the other Tspans, the TspanC8 subgroup (TspanC8s) has eight cysteine residues within the main extracellular region, which form structurally important disulfide bridges for inter-Tspan interactions and regulate ADAM10 activity. The subgroup consists of six members (i.e., Tspan5, Tspan10, Tspan14, Tspan15, Tspan17, and Tspan33) and forms six different "scissor complexes" with ADAM10 that have distinct substrate repertoires. One such notable member of TspanC8s is Tspan15 which has been identified to interact with ADAM10 and suggested as a potent regulator of ADAM10 proteolytic activity. More recently, inhibitory PrP shedding has been observed in Tspan15 knockout mice, suggesting the possibility that Tspan15 causes ADAM10 to release PrP from the cell surface. Although there is currently no definitive evidence to address the dynamic assembly and disassembly of the Tspan15-ADAM10-Q212P complex in Tspan microdomain, it is likely that Tspan15 and/or possibly the other members of TspanC8s modulate ADAM10-mediated Q212P shedding in the complex. Importance of regulation in cell and organism Given the vulnerability of Q212P to environmental redox perturbations, catalysis of sheddase ADAM10 appears to be a "double-edged sword" that, on the one hand, protects the cell surface from Q212P, but on the other, spreads misfolded Q212P to neighboring cells. These opposing reactions must be fine-tuned via ADAM10 in a timely and cooperative manner, otherwise, Q212P accumulates either on the cell surface or in the extracellular space. Although we did not perform an in-depth investigation on the intrinsic properties of Q212P, our analyses in ADAM10 cells provided several valuable clues for a better understanding of the pathological/physiological significance of Q212P shedding mediated by its catalytic interaction with ADAM10. First, inhibitory Q212P shedding was observed as pro-proliferative. This effect was seen in ADAM10 cells expressing Q212P and confirmed by an increase in the size rather than the number of viable colonies grown from single cells (Fig. 6A, see also Fig. S6). In addition, it was not surprising that the cell surface accumulation of Q212P mediated by inhibitory shedding did not induce proteotoxicity, as it was determined to be similar to wtPrP in terms of detergent solubility and protease sensitivity (Fig. S5A), the two typical indicators of proteotoxicity for PrP. Upon release, the extracellular Q212P was expected to be deleterious on its own, as shown by its reduced solubility in the CM, but was diluted to ineffective concentration in the extracellular space, therefore, making it difficult to observe overt proteotoxicity. In contrast, while on the cell surface, Q212P only affected surrounding cells, it was rapidly endocytosed into lysosomes for degradation before exerting proteotoxicity. This might be a plausible explanation for the increased size of the viable colonies as a result of inhibitory Q212P shedding (Fig. 6). Second, ADAM10 served as a negative regulator of pmQC. This fact may be reasonable owing to Q212P's tendency to shed extracellularly in contrast to PrP*, a well-known pmQC substrate that is prone to rapid lysosomal degradation. Nonetheless, both Q212P and PrP* were assumed to be RESET substrates that were rapidly delivered to the cell surface from the stressed ER. During RESET, Q212P formed a complex with precursor ADAM10 (Fig. S4A), while PrP* is known to interact with TMED10. Thus, ADAM10 regulates the Q212P with two sequential functions with the precursor ADAM10 serving as a chaperone that shields misfolded Q212P during the post-ER itinerary, and mature ADAM10 subsequently acting as a sheddase that mediates the proteolytic release of surface Q212P into the extracellular space. Therefore, it can be concluded that ADAM10 participates throughout the secretory itinerary of Q212P, including RESET and pmQC, and allowed PrP accessing the cell surface via RESET to bypass pmQC and get released into the extracellular space (Fig. 7). From a larger perspective, ADAM10 was worth considering as a proteostasis checkpoint to ensure the safety of the secretory and endocytic pathways from misfolded PrP (discussed further in Supplementary Note). Last, ADAM10 deserves significant consideration as a valuable therapeutic target for various human diseases. Besides PrP, more than 100 other substrates, including Notch receptor, APP, and cadherin family, are processed by ADAM10. These substrates are important regulators of cell adhesion and receptor signaling, and their dysregulations are often associated with pathological progression in human diseases, such as brain disorders including Alzheimer's disease, prion disease, fragile X-syndrome, Huntington's disease; cancer including T cell acute lymphoblastic leukemia, glioblastoma, breast cancer, squamous cell carcinoma; and immune disorders including psoriasis, systemic lupus erythematosus, rheumatoid arthritis, Staphylococcus aureus infections, chronic liver inflammation, atherosclerosis. The role of ADAM10 substrates in these disease progressions suggests the potential of ADAM10 as a promising therapeutic target. For instance, Alzheimer's disease is characterized by deposition of pathogenic amyloid peptide (A) derived from amyloid precursor protein (APP), an ADAM10 substrate. ADAM10 activation has been expected to inhibit disease progression in Alzheimer's disease. In addition to promoting ADAM10 activity, the therapeutic strategies aimed towards inhibiting its activity by targeting its active site have been tested in the field of synthetic small molecules. Several hydroxamate inhibitors that chelate the Zn 2+ of the ADAM10 active site, including GI254023X, have been screened and developed. However, despite their high potential demonstrated through the cell and animal-based experiments, they are frequently withdrawn from clinical trials owing to insufficient selectivity, toxicity, and drug resistance. These side effects may be attributed to the varieties of substrates and expression patterns of ADAM10, necessitating a better understanding of the functional consequences of various substrates shed by ADAM10. This requires intensive investigation of the expression, regulation, localization, and interacting partners of ADAM10 in cell culture and animal models. Declaration of competing interest The authors declare that no competing interests exist. Data availability Data will be made available on request.

Phosphatidylethanol Formation via Transphosphatidylation by Rat Brain Synaptosomal Phospholipase D Abstract: Phosphatidylethanol (Peth) formation catalyzed by the transphosphatidylation activity of phospholipase D was demonstrated to occur in a rat brain synaptosomal enriched preparation. The optimal pH was determined to be 6.5, and the optimal ethanol concentration was determined to be 0.30.4 M with an apparent Km of 0.2 M. Peth formation was barely detectable in the absence of an appropriate activator and several unsaturated fatty acids were found to be effective activators. The concentrations of oleic acid required for maximum activation varied with the concentration of exogenous phosphatidylcholine present in the incubation mixtures. All detergents tested were significantly less active than the unsaturated fatty acids and divalent ions were not required for Peth formation. Phosphatidylcholine was the most effective phosphatidyl donor of the phospholipids tested. Peth forming activity was greatest in the synaptic membrane fraction of the various brain subfractions examined. The 12,000 g100,000 g paniculate fraction of lung, heart, and adipose tissue had activities similar to that of brain.

Seniors who have taken an active role find themselves more functionally independent then those who did not. On a fitness scale, it seems to appear seniors in their 60s and 70s these days are unlike those of the previous generations. As we move into a time when there is a greater number of individuals over age 60 than ever before, along with the progression of wellness over the last 20 or so years, I think we need to redefine the meaning of senior fitness. Seniors in their 60s and early 70s who choose to take an active role in their wellness during their 40s and 50s or earlier and stayed the course, find themselves more functionally independent than those who did not. In years past, fitness for seniors was viewed as basic movements and resistance that helped maintain general active daily living. As long as there is no limiting pathology or injury, the functional level of an individual can always improve to higher than above average levels. Challenging one's physical capacity also increases mental cognition along with slowing the aging process. From a functional exercise standpoint, seniors in their 60s have the same capability as individuals in their 40s. There does not have to be limitations to your level of fitness once you are over 60. You truly still have the physical capacity to reach new levels of fitness and not be limited to general levels of exercise deemed for seniors. "My generation embraces the values of exercise and a healthy diet," Sherry Carvalho said. "I believe people now understand that age is just a number and they can have an impact on their quality of life and longevity. As we age we should be consistent in challenging, improving, and maintaining a high functional level of life. This is achieved through purposeful exercise and activity. Physical independence becomes so important as one advances into their senior years of life. "I would encourage all able seniors to challenge their current level of balance and flexibility in order to prevent the falls and injuries that plague us as we age," said 68-year-old Brenda Thompson. "I encourage seniors to participate in goal-oriented programs with the assistance of a professional who can help ensure success and prevent injury. The investment of time and money is truly worth it." Sherman Rosier is the owner of Fit & Functional gym on Thomasville Road.

Application of innovation-based genetic control scheme to signal acquisition for global navigation satellite systems This paper utilizes a novel evolutionary approach to proceed with signal acquisition in Global Navigation Satellite Systems (GNSS). The method controls search space bound of frequency shift and code delay to shorten signal acquisition time. Simulation results show that the proposed scheme can acquire more accurate signal parameters and take less signal acquisition time than the traditional method.

Analytical study of the edge states in the bosonic Haldane model We investigate the properties of magnon edge states in a ferromagnetic honeycomb spin lattice with a Dzyaloshinskii-Moriya interaction (DMI). We derive analytical expressions for the energy spectra and wavefunctions of the edge states localized on the boundaries. By introducing an external on-site potential at the outermost sites, we show that the bosonic band structure is similar to that of the fermionic graphene. We investigate the region in the momentum space where the bosonic edge states are well defined and we analyze the width of the edge state and their dependence with DMI strength. Our findings extend the predictions using topological arguments and they allow size-dependent confirmation from possible experiments I. INTRODUCTION Perhaps the most important and intriguing aspect of the topological insulators is the presence of edge states. A well known example is the Kane-Mele model in graphene, where the spin-orbit coupling (SOC) causes a transition from a semi-metal to a quantum spin Hall insulator. Accompanying the transition is the appearing of gapless edge states with distinct properties from the bulk energy band. In a fermionic model, the edge states are spin-filtered, since the electrons with opposite spins propagate in opposite directions with robustness against external perturbations. The edge states have also been studied in magnetic insulators, where the spin moments are carried by magnons. The magnon Hall effect was observed in a collinear ferromagnetic insulator Lu 2 V 2 O 7 with pyrochlore structure, in the Kgome ferromagnetic lattice, and have also been studied in the Lieb lattice and honeycomb ferromagnetic lattice. Until recently, most investigations of the edge states have been based on fermionic models. Now similar study has been extended to the bosonic models. For example, it has been recently shown that the bosonic equivalent for the Kane-Mele-Haldane model is a ferromagnetic Heisenberg Hamiltonian with the Dzialozinskii-Moriya interaction. The thermal Hall effect and spin Nernst effect have been predicted for this magnetic system. So far, the study of the magnon edge states is mainly based on topological arguments and clearly the detailed properties of the edge states will be required if the potential of magnonics is to be realized. In particular, systematic investigation of the spin-density profile (or magnon density) of the edge magnon and their dependence with the DMI strength and external on-site potentials may be useful in manufacturing small-sized devices based on such magnets. Although a direct experimental demonstration of the magnon edge modes in magnetic systems is still absent, it is interesting to ask whether the similar edge properties in the fermionic systems are also exhibited in the bosonic counterparts. Particularly, with regard to the fact that the strength of DMI in some magnets is greater than that of SOC in graphene by five orders of magnitude. With such motivations in this paper we analyze a ferromagnetic honeycomb lattice with DMI. After introducing an on-site potential at the outermost sites we derive an analytical form for the wavefunctions and energy spectrum for the edge states. We investigate the conditions in the momentum space where the edge states are well defined and we write the edge-state width in terms of the DMI strength. This paper is organized as follows: In Sec. II we introduce the model Hamiltonian with a next-nearest neighbor DMI which opens a gap in the energy spectra. In Sec. III we adapt the analytical approach for the edge state developed by Wang. et. al. and Doh. et. al to a bosonic Haldane model. In order to cover the region where most study has been focusing using topological approach, we introduced an on-site potential at the outermost sites. In Sec. IV, the edge states and their energy spectra for zero and non-zero DMI are investigated. We also analyze the range in the momentum space in which the edge states are well defined and we express the width of the edge state in terms of the DMI strength. We conclude with a summary in Sec. V. A brief description to obtain the edge states and the energy spectrum without DMI is presented in the appendix. II. MODEL HAMILTONIAN We consider a ferromagnetic Heisenberg Hamiltonian with DMI on a honeycomb lattice. The DMI vanishes for the nearest-neighbors (NN) but has non-zero component along the z−direction for the next-nearest-neighbors (NNN). Assuming − → D = D, the corresponding model Hamiltonian for J > 0 is arXiv:1704.07746v3 9 Apr 2019 where i, j indicates the NN coupling, i, j is the NNN coupling, in analogy with the Kane-Mele model, and ij = ±1, depending on the orientation of the two sites. The energy spectrum of Eq. is given by in the linear spin-wave approximation for the infinite system. Here, k = 1 at the two Dirac points K and K in the spin-wave spectra and edge states emerge with gapless energy. Motivated by such predictions, in the following sections, we derive analytical expressions for the edge states and explicit forms for the energy spectrum for the semi-infinite system. zig-zag edges., are the nearest-neighbors and the next-nearest neighbor index vectors, respectively. ij = ±1 is the orientation dependent coefficient and is positive (negative) if the electron makes a left (right) turn to get the second site. The external on-site potential = 1 is introduced at the outermost sites. A. Hamiltonian matrix elements Starting from a Nel ordering state for a bipartite lattice and by the standard Holstein-Primakoff transformations in the linear spin-wave theory, we can write the following effective bosonic Hamiltonian for a ferromagnetic lattice of Eq. as, where the index i (j) denotes the A (B) sublattice sites, and are the NN and NNN vector, respectively, as shown in Fig.. This Hamiltonian is the bosonic equivalent to the Haldane model. To investigate the edge state, we assume a zigzag boundary along the x direction and semi-infinite in the y direction. If k is the momentum in the x direction, the Hamiltonian can be written as, where 2 k, D = D/J, and T is a semi-infinite displacement matrix as defined in Ref.. Here, we introduced an external on-site potential,, at the outermost sites. In a ferromagnetic lattice the intrinsic on-site potential is given by the number of nearest-neighbors, along the zig-zag edge there is a missing bond and the edge on-site energy is lower than in the bulk. By introducing an external onsite potential the edge state can be modified. In this work, we will focus on the "symmetric" case with = 1, as in Refs., where the on-site potential along the edge is the same as in the bulk. We will consider other cases with different values of elsewhere. B. Self-consistent equation for the edge modes From the Hamiltonian and with the explicit form of the matrix elements, we can construct the coupled Harper equations for the edge state. Using the assumption that the edge states are exponentially decaying from the boundary, we consider the following anzats for the eigenstates of M in Eq., In the above wavefunction k is a 2−component vector, z is a complex number with magnitude less than the unity and n = 1, 2, 3..., are the lattice coordinates in the y direction as shown in Fig.. The form z n in the above equation is a finite-size Fourier transform along the ydirection and is related to the eigenvalues of a finite-size displacement matrix. Therefore, the effective Hamiltonian for the edge state can now be written as, The characteristic equation for the eigenvalue is obtained from the condition, |H (z) − | = 0, or, where, a = −J 2 We solve for z + z −1 to obtain four solutions z ± 1 and z ± 2 in terms of the momentum k and energy. Two of the solutions with |z| < 1 describe the state at the upper edge, while the other two with |z| > 1 describe the state for the lower edge, if any. The eigenfunction of Eq. satisfying lim n→∞ k (n) = 0 may now in general be written as, where u 1 are normalization constants and the eigenvector of the Hamiltonian, Eq., corresponding to the solution z r (r = 1, 2), is given by If an opposite boundary exists, the semi-infinite matrices in Eq. can be truncated at some size N and the edge states of the system will become a linear combination of the solutions at each boundary. As N increases over some value, the edge states become independent of the size as expected, allowing us to obtain the solutions at each edge independently. For a large N and for the opposite boundary, the edge state can be obtained if we perform the substitution z n → z N −n+1 in the Eq.. Furthermore, we have to choose the eigenvector whose A−sublattice component goes to zero when D → 0 (the details of the solution without DMI are presented in the appendix). From the Hamiltonian, the eigenvector, r, for the opposite edge, has the form, For a zig-zag edge, the boundary conditions are satisfied by setting: As in the Kane-Mele model, the eigenvectors for each solution are linearly dependent. Then, from the Eq. and the edge condition, Eq., the non-trivial solution is given by setting the determinant | 1 2 | = 0 or | 1 2 | = 0 for the other edge, providing us the following additional relation for the edge energy dispersion, with eigenstates for the upper edge. As we mentioned before, for the lower edge at large N, the eigenstate and energy spectrum can be obtained by the substitutions z n → z N −n+1 in the Eq. and z → z −1 in the Eq.. A. Energy Spectrum Equations and can be solved numerically for the energy and the wavefunction of the edge state. We first discuss the case of D = 0. As shown in Fig. 2(a), the bulk energy spectra is always degenerate at the K and K points, independently of the boundary conditions. However, if we consider a semi-infinite lattice with zig-zag edges, a flat band appears (see appendix for further details). Such flat band is the bosonic analogous to the edge state in zig-zag graphene and the corresponding edge state can be described in a similar way as follows: the bulk bands for a ferromagnetic lattice are symmetric around = 3, hence, if a state exists along the boundary, the sum of the components of the wavefunction over the nearest-neighbors sites must be equal to the on-site energy. Such equality is only fulfilled if we include an external on-site energy, in particular for = 1 we obtain an edge state localized at the A−sublattice. From the Eq. (A5), we can write the wavefunction as, where J 1 is a function of k as we defined before. Hence, the spin-density of the edge magnon is proportional to J 2(n−1) 1 in the region 2 3 (between the K and K points) where the flat band exists. At the point k 0 = √ 3 the edge state is completely localized at the single line of boundary sites and gradually extends inside to the inner sites as we move to the Dirac Points where it merges into the bulk. For the opposite edge, with the same argument, an edge state exists and is localized at the B−sublattice. We next consider the case of non-zero DMI. The energy bands are obtained by solving the Eq. with the solutions given by the Eq., and the edge wavefunction by the Eq.. In the Fig. 2(b) we show the energy bands for a DMI strength of D = 0.1 J. The shaded regions correspond to the bulk spectra, while the two bands which transverse the gap, connecting the upper and lower bulk bands, are the spectra of the edge states. By the analysis of the Eq. or the wavefunctions of Eq., we find that there is only one edge state at each boundary. The edge band at the upper boundary has negative slope while the edge band at the opposite edge has positive slope. Hence, as predicted in Ref., the magnons are moving to the left at one of the edges, while they are moving to the right in the other one. On the other hand, at the point k 0 = √ 3 and = 3, the edge state for the upper edge takes a simple form, The spin-density of the edge magnon is therefore proportional to |z 0 | 2(n−1) and |z 0 | 2n for the A and B sublattice, respectively, decaying exponentially. As we move from k 0 to near the Dirac points K or K, the edge state becomes delocalized and merged with that of the bulk state. B. Allowed range for the edge states We notice that the restriction |z 1,2 | < 1 in the Eq. and allow us to investigate two important properties of the edge states: the region in the momentum space where they are well defined and their confinement to the physical boundary. In general, a particular edge state has two critical points defined as k l and k r located before and after the two Dirac K and K points, respectively, as indicated in the Fig. 2(b). For the upper edge, the eigenfunctions (and also the energy spectrum) depends on z + 1 and z + 2 in the (left) interval k ∈ (k l, k 0 ) and on z − 1 and z − 2 in the (right) interval k ∈ (k 0, k r ). The critical points are determined when the modulus of one z in Eq. reach the unity and the edge state becomes indistinguishable from the bulk. In the Fig. the plot of |z| versus k is shown for different values of D in the interval where the edge state is well defined. For the edge state at the upper edge, the critical point near K is given when the modulus z + 2, reach the unity, whereas that in the right interval (near K ), the critical point is given when z − 1 = 1. In particular for D = 0.1 J, k l ≈ 1.09 and k r ≈ 2.53 . Only when D = 0, these critical points correspond to the Dirac points. If we increase the value of D, the critical points move further away from the Dirac points as shown in Fig.. This suggests that the edge state merges into the bulk but remains in (or near) its boundaries. Such critical points are important since they define the range where the edge state is well defined and they also determine the critical region where z is transformed from real to a complex with unitary modulus. The z solutions with unitary modulus, which obey the bulk and boundary conditions, are sensitive to the values of DMI and the details of edge modifications. C. Width of the bosonic edge state In the previous section, we have investigated the critical points and their dependence with the solutions z + 2 and z − 1 in their corresponding intervals. In this section, we will examine the width of the edge state and their relation with the remaining solutions, z + 1 and z − 2. As described before, the spin-density of the edge magnon decreases as we move away from the boundary, and this happens with some characteristic length scale. Following the Ref. for a semi-infinite zig-zag graphene, the width or characteristic length scale is given by, where |z i (k)| < 1 is the smallest factor in the eigenstate given by Eq.. We notice that such condition is fulfilled by z + 1 and z − 2 between (k l, k 0 ) and (k 0, k r ), respectively, as shown in Fig.. In Fig. we plot the width versus momentum for different values of D, where by completeness we also have included the larger decaying factors. In the slightly flat region around k 0 the width is almost constant. Here, z + 1 = z + 2 and z − 1 = z − 2, since the discriminant of the Eq. is negative (b 2 −4ac < 0), which gives rise a two pairs of complex conjugate solutions, one pair with magnitude lower than one. However, out of this complex region and between the critical values, the solutions are real. In such region, the width associate to the solution z + 1 decreases while the associate to z + 2 grows quickly and diverges at the critical point where the edge state merges into the bulk. Since the edge state wave-function is written as a linear combination, Eq., then the edge state merges into the bulk through a bifurcation of the edge state width in complete analogy with a semi-infinite graphene. Expanding near D = 0 the above equation takes the form, which shows that the width of the edge state vanishes logarithmically as the DMI approaches to zero. Such behavior can be also observed in the Fig., where the z solutions collapses to a single, z = −J 1. In consequence, at D = 0 and k = k 0 the edge state is completely localized at the single line of boundary sites as we mentioned before. As in the fermionic model, the width of the edge state as we discussed above could have important effects when determining the edge properties of samples with finite size. For example, it has been found that there is magnon propagation in interfaces between ferromagnets and normal metals with a strong spin-orbit coupling, referred to as interfacial DMI interaction. It also have been proposed magnon waveguides, where the propagation is between the interface of two topological insulators. In both such cases, the width of the edge state will be useful in the measurement of the magnon current. V. CONCLUSIONS We have derived analytical expressions for the magnon edge-state wavefunctions and their energy spectra for a honeycomb lattice with DMI and zig-zag edges. We demonstrate that the bosonic edge states are defined in a region in the momentum space before merging into the bulk and the width of the edge state can be controlled by the DMI strength. By introducing an on-site potential at the outermost sites we found that the magnonic ferromagnetic band structure closely resembles that of the fermionic graphene. However, unlike graphene where the energy spectrum is doubly degenerate, our explicit solutions demonstrate that there is only one chiral magnon edge state at each edge as predicted by topological approaches. Moreover, our approach is applicable for arbitrary values of the on-site potential as introduced in Eq.. The edge-magnon width, the chirality and the spin density as a function of the DMI presented in this paper could be useful for experiments in small sized monolayers or thin film magnets. by Mexico's National Council of Science and Technology (CONACYT) under the scholarship No. 381939.

MISOPROSTOL STIMULATES cAMP GENERATION IN HUMAN LEUKOCYTES: SYNERGY WITH COLCHICINE SUGGESTS A NEW POTENTIAL FOR ESTABLISHED DRUGS In isolated leukocytes, elevation of cAMP can inhibit various proinflammatory and immune functions. The prostaglandins E (PGEs) are known to stimulate leukocyte cAMP production, and for years they have been viewed as potential immunosuppressive and/or anti-inflammatory agents. However, their clinical use is severely limited by extreme metabolic instability and by poor oral absorption, which necessitates administration by infusion or injection. Misoprostol is a synthetic analog of PGE1 that is relatively stable and orally absorbable. We examined the effects of misoprostol on cAMP production in leukocytes, in view of the possibility that it mimics PGE1 and, thus, might represent a clinically useful immunosuppressive or anti-inflammatory drug. Our results indicate the following: Misoprostol increases leukocyte cAMP production in a dose-dependent manner (∼ 20 nM to > 100 M) and acts by stimulating adenylate cyclase. Its potency and maximal effect are somewhat less than those of PGE1 cAMP generation in response to either misoprostol or PGE1 is transient (in the presence of isobutylmethylxanthine to inhibit endogenous phosphodiesterases). Misoprostol's stimulation of adenylate cyclase is synergistically increased by pretreatment of cells with colchicine, a microtubule-disrupting agent that is currently used for prophylaxis and treatment of gout. Colchicine acts by increasing the initial rate of cAMP production and not by prolonging the response to misoprostol. A clinically relevant dose of colchicine (0.25 M) is effective given sufficient pretreatment time. Whereas a clinically relevant dose of misoprostol (3 nM) is ineffective alone, preexposure of cells to colchicine enables such a dose to stimulate cAMP generation significantly. The combination of misoprostol with colchicine might eventually prove useful in the therapy of immune or inflammatory disease.

A cloud robot system using the dexterity network and berkeley robotics and automation as a service (Brass) In support of Cloud Robotics, Robotics and Automation as a Service (RAaaS) frameworks have the potential to reduce the complexity of software development, simplify software installation and maintenance, and facilitate data sharing for machine learning. In this proof-of-concept paper, we describe Berkeley Robotics and Automation as a Service (Brass), a RAaaS prototype that allows robots to access a remote server that hosts a robust grasp-planning system (Dex-Net 1.0) that maintains data on hundreds of candidate grasps on thousands of 3D object meshes and uses perturbation sampling to estimate and update a stochastic robustness metric for each grasp. Results suggest that such a system can increase grasp reliability over naive locally-computed grasping strategies with network latencies of 30 and 200 msec for servers 500 and 6000 miles away, respectively. We also study how the system can use execution reports from robots in the field to update grasp recommendations over time.

S. Korea, US to work together against outer space waste By Kim Hyo-jin Korea and the United States have agreed to jointly take measures against space debris in response to the rising risk of possible collisions with satellites. Officials from both governments held their first annual talks on space policies at the U.S. State Department in Washington D.C. Thursday and shared concerns about the increasing space waste. The agreement came days after a South Korea satellite barely avoided a collision with a piece of debris produced by the 2009 collision between an inactive Russian communications satellite and an active U.S. satellite. “Securing stability in space is becoming more important as there is an increasing amount of space debris that can possibly collide with satellites,” said a foreign ministry official, adding that both nations shared the necessity of jointly responding to the issue. During the talks, they discussed joint measures to raise “Space Situational Awareness.” This refers to the ability to view, understand and predict the physical location of natural and manmade objects in orbit around the Earth. Both parties are expected to hold a second dialogue sometime next year.