In the present study, a participation of RhoA-mediated Ca2+ sensitization in the augmented bronchial smooth muscle (BSM) contraction in a murine model of allergic asthma was examined.It has recently been suggested that RhoA plays an important role in the enhancement of the CaOvalbumin (OA)-sensitized BALB/c mice were repeatedly challenged with aerosolized OA and sacrificed 24 hours after the last antigen challenge. The contractility and RhoA protein expression of BSMs were measured by organ-bath technique and immunoblotting, respectively.+-depolarization. In α-toxin-permeabilized BSMs, ACh induced a Ca2+ sensitization of contraction, which is sensitive to Clostridium botulinum C3 exoenzyme, indicating that RhoA is implicated in this Ca2+ sensitization. Interestingly, the ACh-induced, RhoA-mediated Ca2+ sensitization was significantly augmented in permeabilized BSMs of OA-challenged mice. Moreover, protein expression of RhoA was significantly increased in the hyperresponsive BSMs.Repeated OA challenge to sensitized mice caused a BSM hyperresponsiveness to acetylcholine (ACh), but not to high K2+ sensitizing effect, probably via an up-regulation of RhoA protein, might be involved in the enhanced BSM contraction in antigen-induced airway hyperresponsiveness.These findings suggest that the augmentation of Ca Increased airway narrowing in response to nonspecific stimuli is a characteristic feature of human obstructive diseases, including bronchial asthma. This abnormality is an important symptom of the disease, although the pathophysiological variations leading to the hyperresponsiveness are unclear now. Several mechanisms have been suggested to explain the airway hyperresponsiveness (AHR), such as alterations in the neural control of airway smooth muscle , increas2+ concentration in myocytes. Recently, additional mechanisms have been suggested in agonist-induced smooth muscle contraction by studies in which the simultaneous measurements of force development and intracellular Ca2+ concentration, and chemically permeabilized preparations in various types of smooth muscles were used. It has been demonstrated that agonist stimulation increases myofilament Ca2+ sensitivity in permeabilized smooth muscles of the rat coronary artery  = 5.67 ± 0.04, Emax = 26.7 ± 1.2 mg; N = 6) and repeatedly OA-challenged (pEC50[Ca2+ (M)] = 5.78 ± 0.15, Emax = 22.8 ± 5.9 mg; N = 6) groups. In both groups, when the Ca2+ concentration was clamped at pCa 6.0, application of ACh (10-5–10-3 M) in the presence of GTP (10-4 M) caused a further contraction, i.e., ACh-induced Ca2+ sensitization, in an ACh concentration-dependent manner . However, the ACh -induced Ca2+ sensitizing effect was inhibited by treatment with C3 in both the sensitized control and OA-challenged groups . In the Pro-Q Diamond dye-stained gels, there were several positive bands, i.e., phosphorylated proteins [bottom panel). The ACh-induced phosphorylation of MLC in BSMs of OA-challenged mice was markedly augmented as compared with those of control animals. A Pro-Q Diamond dye-positive 140 kD band probably corresponding to MYPT1, i.e., phosphorylated MYPT1, was also found in the ACh-stimulated BSM samples and was increased in the OA-challenged group (data not shown).Figure proteins , in the in vivo AHR accompanied by increased IgE production and pulmonary eosinophilia has been demonstrated in the actively sensitized and repeatedly OA-challenged BALB/c strain of mice [+-depolarized (without receptors stimulation), intact muscle strips of the repeatedly OA-challenged mice . These findings suggest that the C3-sensitive, RhoA-mediated Ca2+ sensitization might be augmented in BSMs of the OA-challenged AHR mice. Indeed, the current study also demonstrated a marked increase in the expression and activation of RhoA protein in BSMs of the AHR mice  of α-toxin-permeabilized BSMs was observed between groups (see Result section), indicating that the contents of typical contractile elements such as calmodulin, myosin light chain  BSMs induced by high K+ depolarization was not changed after OA challenge also support our speculation. Thus, the baseline Ca2+ sensitivity of contractile elements themselves in BSM cells is unlikely to change in AHR.In the present study, no significant difference in the Ca2+ itself, the ACh-stimulated contraction of intact BSM strips from the OA-challenged mice was significantly augmented as compared to that from the sensitized control animals  and receptor-operated (nicardipine-insensitive) Ca2+ channels [2+ mobilization in BSM cells. However, the possibility might be denied by the current result that the ACh-induced contraction of α-toxin-permeabilized BSMs from the OA-challenged mice was significantly augmented as compared with that from the control animals even at a constant Ca2+ concentration  of up-regulation of RhoA in OA-challenged BSMs is not known here, inflammatory cytokines such as tumor necrosis factor-α [One of the important findings in the present study is that the C3-sensitive, RhoA-mediated Cace Figs. . Thus, tfactor-α , which ifactor-α ,35. It i2+ level was within normal level even in the hyperresponsive BSMs [2+ sensitization, although the mechanism(s) for activation of RhoA by ACh is still unclear. If RhoA proteins are activated by receptors other than muscarinic receptor, it might account for the 'non-specific' AHR, which is a common feature of allergic asthmatics. Indeed, the BSMs of the OA-challenged mice also have hyperresponsiveness to endothelin-1 [An increase in responsiveness to muscarinic agonists of airway smooth muscle has been demonstrated in animal models of AHR ,22,36,37ive BSMs ,30. Takethelin-1 , which hthelin-1 .An upregulation of RhoA/Rho-kinase associated with the augmented smooth muscle contractility has also been reported in rat myomertium during pregnancy ,43, arte2+ sensitization in murine BSM contraction. An augmentation of the Ca2+ sensitizing effect, probably by the upregulation of RhoA protein, might be involved in the enhanced BSM contraction observed in the antigen-induced AHR in mice.In conclusion, the current study demonstrated an ACh-induced, RhoA-mediated CaYC conceived of the study, participated in its design and coordination, and drafted the manuscript. AU carried out the intact smooth muscle studies. KS, HT and HS carried out the skinned fiber studies and immunoblot analysis. SN carried out the analysis of active RhoA. MM participated in the direction of the study as well as writing and preparing the manuscript. All authors read and approved the final manuscript.


The brain-derived protein S100B has been shown to be a useful marker of brain injury of different etiologies. Cognitive dysfunction after cardiac surgery using cardiopulmonary bypass has been reported to occur in up to 70% of patients. In this study we tried to evaluate S100B as a marker for cognitive dysfunction after coronary bypass surgery with cardiopulmonary bypass in a model where the inflow of S100B from shed mediastinal blood was corrected for.56 patients scheduled for coronary artery bypass grafting underwent prospective neuropsychological testing. The test scores were standardized and an impairment index was constructed. S100B was sampled at the end of surgery, hourly for the first 6 hours, and then 8, 10, 15, 24 and 48 hours after surgery. None of the patients received autotransfusion.In simple linear analysis, no significant relation was found between S100B levels and neuropsychological outcome. In a backwards stepwise regression analysis the three variables, S100B levels at the end of cardiopulmonary bypass, S100B levels 1 hour later and the age of the patients were found to explain part of the neuropsychological deterioration .In this study we found that S100B levels 1 hour after surgery seem to be the most informative. Our attempt to control the increased levels of S100B caused by contamination from the surgical field did not yield different results. We conclude that the clinical value of S100B as a predictive measurement of postoperative cognitive dysfunction after cardiac surgery is limited. Despite the fact that incidence figures between 4–79% have been reported for cognitive dysfunction after cardiac surgery , diagnosSeveral studies in humans suffering from stroke of different ethiologies, have shown a rather strong correlation between serum levels of S100B and size of lesion(s) as well as outcome -17.Lately a number of studies have addressed the question whether S100B can be considered as a marker for cognitive dysfunction after cardiac surgery; however the conclusions presented are disparate. One major reason for this could be the fact that S100B is present in high concentrations in shed mediastinal blood that is retransfused to the patient by cardiotomy suction and autotransfusion, thus obscuring the measured levels of S100B early after surgery. To date, only one study have been published where this contamination was taken into account . We receThe study group comprised 56 patients who underwent coronary artery bypass graft (CABG) surgery at the Division of Cardiac Surgery, University Hospital MAS, Malmoe, Sweden and Department of Cardiothoracic Surgery, Lund University Hospital, Lund, Sweden. Only patients planned for elective CABG with cardiopulmonary bypass (CPB) as their sole procedure were included. The study protocol was approved by the local ethics committee, and patients gave a written informed consent before the study protocol was initiated. Patients with a history of stroke, transient ischemic attack (TIA), reversible neurological disorder (RIND), known carotid artery disease or other brain diseases were excluded. In order to avoid possible influence of renal disorder on the elimination of S100B, patients with known renal failure (Creatinin > 160 μmol/L) were excluded.The patients were examined for signs of neurological dysfunctions daily during the hospital stay by either experienced cardiac anesthesiologists or by experienced cardiac surgeons.®, Roche, Basel, Switzerland) or propofol  10 mg/kg. It was subsequently maintained with fentanyl 10 μg/kg , a continuous infusion of propofol 3–6 mg/kg/h or inhalation of isoflurane 0,5–1% . Nitrous oxide  was used before CPB but not during or after CPB.Anesthesia was induced with midazolam 3–5 mg iv  and administered in the ascending aorta and the anastomosed vein-grafts intermittently.CABG surgery was performed during aortic cross clamping with the distal anastomosis preceding the proximal anastomosis. A tangential occluder replaced the cross-clamp during the proximal anastomosis. Antegrade cold S:t Thomas crystalloid cardioplegia was used (Cardioplegi2 at normothermia. The perfusate was cooled to approximately 32°C. Heparin (400 U/kg bodyweight) was given prior to cannulation and reversed with equal doses of protamine sulphate at decannulation.Perfusion was performed with a roller pump . The perfusion catheters and circuit were made of polyvinylchloride in the line and silicon in the pumphead. The arterial cannulation was made in the ascending aorta and venous cannulation in the right atrium by a two-stage venous cannula. All circuits contained a heparin-coated 40 μm arterial filter  and a membrane oxygenator . The circuit was primed with approximately 1000 ml of Ringer's lactate (Pharmacia-Upjohn), 250 ml 15% Mannitol (Pharmacia-Upjohn) and 75 mmol Addex tromethamine (Pharmacia-Upjohn). Perfusion flow was non-pulsatile with a flow rate of 2.4 l/min/mAfter surgery, the patients patient were transferred to the ICU for recovery and enrolled in the sampling scheme for S100B analysis. None of the patients received autotransfusion.Serum for S100B analysis was sampled before surgery, at the end of CPB, and then 1, 2, 3, 4, 5, 6, 8, 10, 15, 24 and 48 hours after surgery. The S100B levels at these time points will be referred to as T0, T1, T2....T48. Blood samples, both arterial and venous samples, were cooled and centrifuged within 5 hours. All samples were measured by a monoclonal two-site immunoluminometric assay .Since early levels of S100B are contaminated by S100B from cardiotomy suction, an attempt was made to exclude this S100B from the levels measured one and two hours after surgery. Assuming that all of the measured S100B at the termination of CPB was a contamination, this non-cerebral S100B was eliminated with a half-life of 25 minutes, as illustrated in figure e is the estimated true levels of S100B at time t, Ct = the measured concentration of S100B at T1 or T2, C0 the concentration at the end of CPB, t the time after T0 and t1/2 the half-life of S100B.where CWe have earlier suggested that the elimination rate could be used as another measure of cerebral release , and theThe patients underwent neuropsychological testing by the same trained neuropsychologist 1–2 days before and 5–7 days after surgery. The tests used were: Mental Control, Figural Memory, Logical Memory (A/B), Visual Reproduction, Rey Auditory/Verbal Learning Test (RAVLT), Trail Making A, Trail Making B, Digit Symbol, Digit Span, Visual Memory Span, Visual Paired Associates II or Verbal Paired Associates I and RAVLT, Delayed Retention. The tests were chosen from the Wechsler Memory Scale-Revised (WMS-R), the Wechsler Adult Intelligence Scale (WAIS – R) and the Halsted-Reitan Neuropsychological Battery ,21.Differences for each sub-test were first calculated and then standardized to z-values. All sub-tests were then aggregated to create an impairment index . The impAll results were analyzed with the Statistica version 5.0 for PC. Regression analysis was performed with the least square method with a casewise deletion of missing data. For the multiple regression a backwards stepwise regression was performed to determine which variables to include in the final analysis. The initial variables included in this analysis were age, gender, perfusion time, years of education, and S100B levels at all sampling times. A p-value < 0,05 was considered significant. Unless otherwise stated numerical values are presented as mean ± 1 standard deviation.Patient demographics are presented in table The appearance of S100B protein followed the same pattern in all patients with high levels at the end of CPB and a decrease or slight increase the first hour figure . ThereafWe found a correlation between patient age and S100B levels measured up to 24 hours after bypass Table . NeitherIn multiple regression analysis, measured S100B levels at the end of CPB (T0), one hour later (T1) and age were found to explain part of the neuropsychological impairment  declare that they have no competing interests.HJ: Principal investigator recruited, enrolled patients analyzed and wrote the paperPJ: Had an integral role in the planning and the analysis, also help with writingMB: Designed neuropsychological test battery. Analyzed neuropsychological dataCB: Performed all neuropsychological testing with patientsCA: Did the S100B-analyzesSB: Mentor an principal leader of the project, who also helped with the writing.The pre-publication history for this paper can be accessed here:


Pulmonary oedema and impairment of oxygenation are reported as common consequences of haemorrhagic shock and resuscitation (HSR). Surprisingly, there is little information in the literature examining differences in crystalloid type during the early phase of HSR regarding the development of pulmonary oedema, the impact on oxygenation and the haemodynamic response. These experiments were designed to determine if differences exist because of crystalloid fluid type in the development of oedema, the impact on oxygenation and the haemodynamic response to fluid administration in early HSR.Twenty anaesthetised swine underwent a grade V liver injury and bled without resuscitation for 30 minutes. The animals were randomised to receive, in a blinded fashion, either normal saline  or lactated Ringer's solution . They were then resuscitated with study fluid to, and maintained at, the preinjury mean arterial pressure (MAP) for 90 minutes.P = 0.027). After 150 ml/kg of fluid, EVLWI increased from 9.5 ± 0.3 ml/kg to 11.4 ± 0.3 ml/kg NS and from 9.3 ± 0.2 ml/kg to 10.8 ± 0.3 ml/kg LR (P = 0.035). Despite this, oxygenation was not significantly impacted /fraction of inspired oxygen (FiO2) ≤ 100) until approximately 250 ml/kg of either fluid had been administered. Animals resuscitated with NS were more acidaemic (with lower lactates), pH 7.17 ± 0.03 NS vs. 7.41 ± 0.02 LR (P < 0.001).Extravascular lung water index (EVLWI) began to increase immediately with resuscitation with both fluid types, increasing earlier and to a greater degree with NS. A 1 ml/kg increase in EVLWI from baseline occurred after administartion of (mean ± standard error of the mean) 68.6 ± 5.2 ml/kg of normal saline and 81.3 ± 8.7 ml/kg of LR (This study suggests that early resuscitation of haemorrhagic shock with NS or LR has little impact on oxygenation when resuscitation volume is less than 250 ml/kg. LR has more favourable effects than NS on EVLWI, pH and blood pressure but not on oxygenation. Pulmonary oedema has been reported to be a common consequence of haemorrhagic shock and resuscitation (HSR) . The earThe EVLWI at any time represents a dynamic balance between factors that cause fluid to accumulate in the lungs and those that carry it out of the lungs. An increase in fluid leaving the vascular space and remaining in the pulmonary parenchyma following HSR can result from: endothelial injury with increased pulmonary capillary permeability; alveolar epithelial injury and decreased alveolar fluid clearance; dilutional decreases in serum oncotic pressure; increased pulmonary capillary pressure; coagulation abnormalities with increased extravasation of fluid; and increased inflammation ,3. But b2)/fraction of inspired oxygen (FiO2) and parameters of haemodynamics during early resuscitation from haemorrhagic shock with LR or NS and determine if there are differences on the development of oedema, the impact on oxygenation and the haemodynamic response to fluid administration because of fluid type.These experiments were designed to measure the EVLWI, the partial pressure of arterial oxygen , intubated with an oral endotracheal tube and placed on mechanical ventilation. Tidal volume was set at 12 ± 2 ml/kg, and respiratory rate was adjusted to maintain end-tidal carbon dioxide and partial pressure of arterial carbon dioxide (PaCO2) of 40 ± 4 mmHg. Anaesthesia was maintained using 1 to 3% isoflurane as needed. To assess adequacy of anaesthesia, we monitored jaw tone. Monitoring devices were placed after establishing anaesthesia, including an oesophageal thermometer and external jugular vein catheter. Animal temperature was maintained at 38.0 ± 1.5°C using external warming devices and warmed fluids. Femoral artery cut down was performed to place a 4-F femoral catheter with an integrated thermistor tip  for continuous blood pressure monitoring, blood sampling and transpulmonary thermodilution determinations of cardiac output (CO), EVLWI, global end-diastolic volume (GEDV) and intrathoracic blood volume (ITBV). MAP and heart rate (HR) were continuously recorded.Twenty Yorkshire crossbred pigs weighing approximately 35 kg underwent a 16-hour fast preoperatively with water The transpulmonary thermodilution method using the single-indicator transpulmonary thermodilution technique  was originally developed in swine and has been previously validated by comparison with the postmortem gravimetric technique, and with the double dilution (thermo-dye) technique in swine, sheep and humans in a variety of disease states -18. The We examined the precision of this technique for measuring EVLWI in this species of swine using repeated measures of EVLWI at baseline in three animals in separate experiments. We found an average coefficient of variation (standard deviation/mean × 100) of less than 5%. As an example, repeated measurements (n = 20) were made in one animal at baseline yielding a mean EVLW (± standard deviation) of 9.1 ± .45 ml/kg.For these experiments, a 15 ml bolus of iced randomised fluid (0 to 6°C) was injected via a central venous catheter into the right atrium. The thermodilution curve was recorded with a femoral artery thermistor and used to determine CO, the volume of blood in the heart at the end of filling or the GEDV, the ITBV and EVLWI, as previously described ,20. The The animals underwent a midline coeliotomy, suprapubic urinary catheter placement and splenectomy. Splenectomies are performed in swine haemorrhage models because of the spleen's distensibility and the resultant variable amounts of sequestered blood. The spleen was weighed and randomised fluid was infused to replace three times the spleen weight. Following a 15-minute stabilisation period, a standardised grade V liver injury was created with a specially designed clamp. The clamp was directed centrally over the liver and created a consistent pattern of injury involving one or more central hepatic veins. Injuries met criteria for grade V liver injuries as described by the American Association for the Surgery of Trauma Organ Injury Scaling system . This moAfter 30 minutes of uncontrolled haemorrhage, blood was evacuated from the abdomen and measured. Blinded resuscitation was begun with either LR or NS. Both fluids were purchased from Baxter  and were unmodified . A 30-minute delay before beginning resuscitation allowed the animals to reach their nadir blood pressure, replicating the civilian trauma scenario. Fluid was delivered at 165 mL/minute. This rate was chosen because it is one-half of the rate delivered by the level 1 infuser to humans, and the pigs were approximately one-half the weight of a normal adult human. The goal of resuscitation was to achieve and maintain the baseline MAP for 90 minutes post injury. Blood loss was determined by placing pre-weighed laparotomy sponges into the pelvis and inferior right and left pericolic gutters before creating the liver injury and collecting active haemorrhage by suction or with the sponges, while avoiding manipulation of the liver injury. The sponges were removed before abdominal closure. Following resuscitation and prior to euthanasia, the abdomen was opened and any additional blood loss was collected by suction. Blood loss (mL/kg) was reported as a mean for each resuscitation group. To ensure comparable injuries between the study groups, we removed the liver and identified the number of hepatic vessels injured.t tests using a statistical software package for personal computers . Significance was defined as p < 0.05. All data are presented as means ± standard error of the mean. Linear regression analysis was performed to determine the effect of fluid type, independent of volume, as shown by differences in the slopes of the regression lines. Boxplots comparing differences in means with confidence intervals of MAP, CO, systemic vascular resistance (SVR), GEDV, stroke volume (SV) and HR were performed to examine differences in those values with NS vs. LR resuscitation.Comparisons between groups were made with independent-sample One animal in the NS group did not survive the two-hour study. All the animals in the LR group survived. Composite MAP of the two groups throughout the study are shown in Figure Mean urine output and blood loss were greater in the NS group than in the LR group. Urine output was 44.1 ml/kg ± 8.1 in the NS group vs. 19.4 ml/kg ± 3.4 ml/kg in the LR group (p = 0.012), and total blood loss was 34.3 ± 2.9 ml/kg vs. 23.7 ± 2.1 ml/kg (p = 0.009). The NS group required almost twice as much fluid  over the 90-minute resuscitation period in an attempt to maintain goal MAP. There was no difference in blood loss prior to fluid resuscitation.2/FiO2 ratio as a function of resuscitation fluid volume over the entire study are plotted in Figure 2/FiO2 of 100  did not occur until approximately 250 ml/kg of either fluid had been administered.EVLWI and the PaO2/FiO2 plotted as a function of resuscitation fluid volume administered early in the study, limited to 250 ml/kg. This was performed to examine the effect of fluid type early in the resuscitation at similar volumes. Linear regression analysis revealed a larger increase in EVLWI in the NS group as compared with the LR group occurring early in the resuscitation. There was no significant difference in baseline EVLWI for the two groups. A 1 ml/kg increase in EVLWI from baseline occurred after 68.6 ± 5.2 ml/kg of NS and 81.3 ± 8.7 ml/kg of LR (p = 0.027). After 150 ml/kg of fluid EVLWI increased from 9.5 ± 0.3 ml/kg to 11.4 ± 0.3 ml/kg for NS and from 9.3 ± 0.2 mg/g to 10.8 ± 0.3 ml/kg for LR (p = 0.035). Significant changes in oxygenation, defined as a drop in baseline PaO2/FiO2 to 100 or greater, did not occur over this range for either fluid type.Figure 3 vs.1177.6 ± 34.6 dyne × sec/m3; p < 0.001). As shown in Figure Figure Figure Laboratory values at the end of the study are shown in Table 2/FiO2 of 100 or more , until approximately 250 ml of either fluid had been infused. This corresponds to 17.5 litres in a 70 kg human – well in excess of what would be normally administered during most human resuscitations prior to the arrival of blood products.In this swine model of traumatic haemorrhagic shock, resuscitation with NS as compared with LR resulted in greater EVLWI formation at similar amounts of resuscitation volume. In addition, the blood pressure response to fluid was considerably lower in the NS group  despite requiring more total fluid over the entire study period  in an attempt to maintain goal MAP. The lower blood pressure with NS administration was due to a greater systemic vasodilatation as compared with LR. Surprisingly neither fluid resulted in significant changes in oxygenation, as defined by a drop in baseline PaOWe observed differences in EVLWI early in the resuscitation when similar volumes of the fluids had been given, more NS than LR. This supports the idea that a fluid specific effect, independent of volume, was present. To evaluate this we performed two separate regression analyses: Figure 2/FiO2 vs. volume of resuscitation for two reasons. First, previous studies have shown a stepwise increase in the rate of EVLWI formation as a function of the interstitial matrix filling and then abruptly 'giving way' with a subsequent increased rate of fluid extravasation into the matrix and then alveoli. One could expect an initial improvement in PaO2/FiO2 with improved CO early in the resuscitation, followed by a curvilinear decline as the matrix 'lets go' with a subsequent increase rate in fluid extravasation. Secondly a curvilinear line dramatically improved the regression coefficient (R2); from 0.620 to 0.815 for NS, and from 0.15 to 0.60 for LR. We found significant differences in EVLWI formation at similar volumes of resuscitation, more NS than LR early in the resuscitation. The cause of the greater EVLWI accumulation in the NS group at similar resuscitation volumes as compared with the LR group is not known.A second order polynomial was used as the best fit line for PaOEVLWI is defined as the extravascular fluid in the lung at any moment including the intracellular fluid of inflammatory, endothelial and epithelial cells in the extravascular space, as well as alveolar and interstitial fluid and intrapulmonary lymph . The amoHSR results in increased expression of inflammatory cytokines, increased neutrophil sequestration in the lung, increased generation of reactive oxygen and nitrogen species, and activation of coagulation. All of these factors can cause endothelial and epithelial cell dysfunction and increased EVLWI ,26-28. HThe NS group developed a non-anion gap metabolic acidosis whereas the LR group remained pH neutral. Several studies have shown potential lung protective effects of hypercapnic acidosis in acute lung injury -32. HoweDilutional decreases in serum oncotic pressure are likely to have contributed to the rise in EVLWI seen with both fluid types in this study. It is likely that the differences in the total EVLWI measured at study end were in large part due to dilutional effects from the greater volume of NS required. Twice the volume of NS was required to maintain target MAP. This has clinical relevance in regards to selecting fluid type, in that a greater volume of NS will need to be administered to maintain blood pressure goals and will result in greater EVLWI formation and acidosis.Figure As shown in Figure Total blood loss was greater in the NS group than the LR group . It is known that HSR with NS attenuates the hypercoagulable response seen with haemorrhage as compared with LR . IncreasPrevious studies examining the effects of NS vs. LR on EVLWI, oxygenation and the haemodynamic response have been conducted in controlled haemorrhage models. Many re-infused shed blood with the resuscitation fluid. Our study used a more clinically relevant model by adding tissue injury to uncontrolled haemorrhage and initiating early resuscitation with crystalloids alone and resuscitating to a goal blood pressure. Although this complicated the analysis of the effect of fluid type independent of volume, we felt by doing so our results could be more reliably extrapolated to the human clinical scenario. Further work is needed to examine late effects of crystalloid type and volume on EVLWI, oxygenation and haemodynamics in a living model of HSR.Our study did have limitations. By study end the animals had received extremely large resuscitation volumes that exceeded those typically seen in human scenarios. We believe this reflects the severity of the model as well as the choice to resuscitate to the baseline MAP. This resulted in resuscitation beyond normal perfusion as evidenced by supranormal urine outputs especially in the NS group. Despite this fact, differences between the fluids manifested very early, prior to exceeding resuscitation volumes typically used clinically. We have also previously shown that differences in pH and coagulation between LR and NS occur very early after initiation of fluid resuscitation . This inA tidal volume of 12 ml/kg was used in this study. This tidal volume may have caused some lung injury. Although this may have played a role in the development of increased EVLWI, both animal groups were exposed to the same tidal volume making it an unlikely cause of a difference between groups.This study suggests that early resuscitation of haemorrhagic shock with NS or LR prior to the arrival of blood products is safe in terms of the immediate impact on oxygenation when resuscitation volume is limited to less than 250 ml/kg. Resuscitation with LR has more favourable effects on EVLWI formation, pH, coagulation and haemodynamics but not on oxygenation. Further work is needed to understand the cause of these fluid-specific differences.• When resuscitating haemorrhagic shock prior to the arrival of blood products significantly more NS is required to maintain goal blood pressure than LR.• LR has more favourable effects on EVLWI formation, pH, coagulation and haemodynamics than does NS.• Further investigation is warranted to attempt to explain these fluid-specific differences.2: fraction of inspired oxygen; GEDV: global end-diastolic volume; HR: heart rate; HSR: haemorrhagic shock and resuscitation; IL: interleukin; ITBV: intrathoracic blood volume; LR: lactated Ringer's solution; MAP: mean arterial pressure; NS: normal saline; PaCO2: partial pressure of arterial carbon dioxide; PaO2: partial pressure of arterial oxygen; PBV: pulmonary blood volume; PVPI: pulmonary vascular permeability index; SV: stroke volume; SVR: systemic vascular resistance; TNF: tumour necrosis factor.CO: cardiac output; EVLWI: extravascular lung water index; FiOCRP has acted as an advisor to Pulsion Medical Systems, makers of the PiCCO device that measures EVLW. There are no other potential conflicts from any of the authors.CRP participated in study design and coordination and helped draft the mauscript. KV helped with data collection and analysis. DSH participated in executing the experiments, and helped with data collection. RSS participated in the design of the study and participated in executing the experiments. JAD participated in the design and coordination of the study, in study execution, data collection and analysis. JMW participated in the design of the study and participated in executing the experiments. MAS conceived of the study, participated in study design and coordination and helped draft the mauscript. All authors read and approved the final manuscript.


Objective: Over the last 15 years, researchers from around the world have developed instruments for assessing the risk of conversion to psychosis. The objective of this article is to review the literature on these instruments by focusing on genealogy links and on their performance in predicting conversion to psychosis.Method: A systematic review of articles published since 1980 relating to risk assessment instruments for conversion to psychosis by manual search and consultation of electronic databases MEDLINE, EMBASE, and PsycINFO.Results: Three hundred ninety one (391) publications were selected and analyzed. Among these, 22 instruments were identified. These instruments are briefly described and placed on a timeline according to their year of publication. A code of positions, patterns, and forms is used to schematize the characteristics of each instrument. A table is presented to show changes in rates of conversion to psychosis within cohorts of subjects considered at risk according to the instruments. A second code of shades and outlines is used to schematize the characteristics of each cohort of patients. The two graphics set the stage for a discussion about the major strategies that were adopted to improve the performance of risk assessment instruments.Conclusion: These graphics allow a better understanding of the origin, evolution, current status, strengths, shortcomings, and future prospects of research on risk assessment instruments.Clinical ImplicationsThe integration of theoretical approaches, the multicenter studies, and the pre-selection of patients with short questionnaires were the main strategies to improve the performance of instruments assessing the risk of conversion to psychosis.These instruments are better at predicting conversion to psychosis than conventional variables within a more limited time span and can therefore enable the evaluation of various risk factors and biomarkers that may be associated with psychosis.LimitationsThe studies selected for this review of literature were not classified according to their methodological quality.These studies are based on heterogeneous populations and this must be taken into account when comparing the rates of conversion to psychosis.This review of literature was based on published data only and they were no direct communication with the authors of these instruments. Beyond its economic implications, psychosis is a disease that all too often has a lasting and painful impact on the patient’s life. In the hope of avoiding the most dramatic consequences, research groups around the world have been working on psychosis prevention since the early 1990s , the development of instruments to evaluate the risk of conversion to psychosis would make it possible to identify individuals who will develop psychosis before the disease sets in and therefore before their lives are disrupted. These people could then receive treatments to mitigate, delay, or even prevent the undesirable consequences. For more than 20 years, such instruments have been developed and tested around the world, starting with the scales created by Chapman and Chapman , who souHowever, the conversion rate in the cohorts of individuals identified as being at risk of developing psychosis are relatively low, ranging from less than 10% to slightly over 50% within follow-up periods ranging from 6 months to 10 years. In many centers, it has also been observed that conversion rates decline with time . The publications retained had to concern one or more instruments for evaluating the risk of conversion to psychosis and to be written in English or French. Particular attention was paid to previously published literature reviews on these instruments to better understand their theoretical bases.To do this, a review of the literature on instruments designed to evaluate the risk of conversion to psychosis was done by our group from November 2009 to July 2011. The keywords The scales and other tools on which they were based were placed on a timeline Figure , and claThe search in PsycINFO returned 240 publications, in EMBASE, 249, and in MEDLINE, 63. After adding the publications found with a manual search and applying the inclusion and exclusion criteria, we retained a total of 391 publications. Of this number, 181 publications (46.3%) came from Europe, 123 (31.5%) from North America, 51 (13.0%) from Australia, 14 (3.6%) from Asia, and 2 (0.5%) from Israel. Certain publications featured contributions by authors from several research centers around the world, and 20 (5.1%) were classified as the outcome of a multicenter cooperative study. Several of these publications concerned the follow-up of cohorts of patients at risk of conversion to psychosis (155 or 39.6%). As well, 71 reviews of the literature (or 18.2% of the publications retained) enabled us to consolidate our knowledge of the topic.Through this search, we identified 22 instruments for evaluation of the risk of conversion to psychosis. These instruments were placed on the timeline based on their publication dates , which occurs at the very start of the development of psychosis , cognitive problems, disorders affecting cenesthesia (the sense of bodily existence), central neurovegetative disorders, and coping strategies  were first operationalized by the research team at the PACE Clinic in Melbourne, VIC, Australia. They refer to three groups of patients: patients who present attenuated psychotic symptoms ; patients who present what are known as Brief Limited Intermittent Psychotic Symptoms or BLIPS ; and patients who present risk factors combined with an at-risk state or trait and state risk factors  . According to this research strategy, patients are at risk of conversion to psychosis if they present attenuated positive symptoms (or CHR+), but also if they manifest attenuated negative symptoms (or CHR−) , was developed by a Japanese team that added a section on the duration of symptoms to improve its specificity (symptoms present for a longer time are considered to be more representative of being at risk of conversion to psychosis)  more specifically indicate an increased risk of conversion to psychosis. In a sample of 501 subjects with at least one DSM-III-R criterion for prodrome, the D3R identified 20 subjects as being at risk of conversion to psychosis; 3 of them (15%) developed a psychosis after 6 months of follow-up  in Switzerland led to the development of several instruments inspired by the DSM-III-R criteria, but also by the literature on prodromal psychosis . The PROD-screen is a 29-question screening questionnaire inspired by the SIPS, the IRAOS, and the BSABS . This is well represented in Figure But this reconciliation of the two approaches is not sufficient to mitigate the differences between the various research teams. Differences remain between the characteristics of the basic population, the recruitment of patients, the evaluation process, the follow-up, the treatments provided, etc. This is why there was the creation of several multicenter studies . In addition to enabling researchers to test evaluation instruments on a much larger population, these studies are pursuing the homogenization of the process of evaluating subjects at risk of conversion to psychosis. This homogenization makes it easier to compare the results of research on these subjects, which had formerly been difficult and risky.Figure We should point out that the way subjects get into programs for evaluating the risk of conversion probably has a great influence on the proportion of subjects who will develop psychosis out of the total cohort of patients considered to be at risk. Yung et al.  hypothesDespite their relatively low positive predictive value, it should be emphasized that these evaluation instruments are better at predicting conversion to psychosis than conventional variables  within a more limited time span Cannon, . They maThese results show how these instruments are evolving. They are constantly being revised as a function of the clinical and ethical challenges represented by research on the prodrome of psychosis. This is what we have attempted to schematize in Figure One of the next steps could be to push the analysis further and to consider changes in these patients’ distress levels and functioning. As Ruhrmann et al.  emphasizThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The first is presented to a scientist who has made distinguished contributions over many years in research, teaching, service, or any combination of the three. This year, the ISCB Accomplishment by a Senior Scientist Award goes to Michael Ashburner in the department of genetics at the University of Cambridge. The second, known as the Overton Prize, honours a young scientist in the early to mid-stage of his or her career who has already achieved significant and lasting impact in the field of computational biology. In 2011, the Overton Prize is awarded to Olga Troyanskaya of Princeton University in New Jersey.Each year, the International Society for Computational Biology  in Madrid. The winners will receive their awards at the ISCB's annual meeting, where they will also deliver keynote talks. This meeting, ISMB/ECCB 2011 If computational biology seems challenging in the second decade of the 21st century, spare a thought for those who pioneered the discipline in the 1980s. Michael Ashburner  at the UDrosophila and, in particular, polytene chromosomes, which form when certain specialised cells undergo repeated rounds of DNA replication. Polytene chromosomes have a characteristic banded structure. In Drosophila there are some 5,000 bands and a subset of these undergo, during development, a reversible structural modification as the result of transcription; this is know as puffing and can be considered an analog of gene activity. In the late 1960s and early 1970s, Ashburner studied puffing patterns and inferred the existence of a cascade of genetic controls under the influence of the hormone ecdysone during larval development.Ashburner began his career with a degree in genetics from the University of Cambridge in 1964. He stayed on to do a PhD, studying Alcohol dehydrogenase gene and its environs. By the mid-1980s, he had the most detailed analysis in full genetic terms of any small chromosome region of any multi-cellular organism, and had the Adh gene sequences from several different species of Drosophila. “That drew me into bioinformatics because we needed a way of comparing sequences,” he says. “There was almost no software available to help.”In the late 1970s, Ashburner turned his attention to the study of the Two people came to his aid. The first was Walter Bodmer, director of the Imperial Cancer Research Fund, who gave Ashburner the use of a DEC computer with access to the early network. “We could access this machine by dial-up and do some analysis,” he says. The second was Doug Brutlag at Stanford University, who was developing MOLGEN, an early bioinformatics system, which he allowed Ashburner to access.That presented a significant obstacle, however. Getting a computer in the United Kingdom to speak to one in Stanford was not straightforward. Today, everybody uses the Internet, defined by the TCP/IP protocol. But in the early ‘80 s, the UK and United States used different systems. The US was pioneering TCP/IP while the UK had a standard called the Coloured Book protocols. “The only place that had an interface between the two protocols was University College, London, and they were very helpful,” says Ashburner, “giving us 5 kb of disk space.”The process of connecting to Stanford was far from simple. “The way you did it was to dial up your local packet switching exchange at the Post Office and connect to the Rutherford Appleton Laboratory. You then typed in some code which connected you to UCL where you could use TCP/IP,” he says. The signal was routed via Goonhilly satellite station in Cornwall to Carnegie Mellon University and from there to Stanford. “I had a dumb terminal, that is a box with no memory, so everything had to be captured by a printer in parallel.” Ashburner was far from deterred, however.At about that time, the European Molecular Biology Laboratory (EMBL) in Heidelberg and GenBank in the US released the first nucleotide sequence libraries in quick succession. Using his network access, Ashburner and his colleagues, collaboratively with MOLGEN, set up one of the first bulletin boards, called BioNet, to keep people informed of changes to the library and to software. “This became well used and things evolved from there,” he says.As the field of bioinformatics grew, the need for an institution to house the data and conduct research increased. So in 1992, the EMBL decided to set up an institute of bioinformatics that would house this library and carry out research. This organisation became known as the European Bioinformatics Institute, based in Hinxton, UK, with Ashburner and John Sulston having led the UK bid to host it. “I was persuaded to become the first program coordinator and took half-time leave from Cambridge to do that,” he says. He eventually took over as joint-director, a post he held until 2001. “At first, the finances were sticky and the politics were horrendous. But it has since gone from strength to strength,” he says.Drosophila genetics. This is a field with a rich and long history of collecting and sharing mutations. The first catalogue of mutations was published in 1925 and it was still being revised in paper form in the late 1980s. But the field was beginning to expand quickly and the books were out of date as soon as they were published. “It became clear to me that we couldn't carry on publishing in paper form every 10 or 20 years,” he recalls.At the same time, Ashburner continued his interest in So in 1989 he proposed that the community set up an electronic database to take over the role of the printed one. In 1992, the NIH funded the project that became known as FlyBase, one of the first genetic and now genomic databases.FlyBase was a crucial factor in triggering Ashburner's interest in a structured, controlled vocabulary, a formal representation of knowledge about genes and gene products. He began to define terms for gene products by their biological processes, such as wing development, and then defined the data structure in which these terms were related to each other. “It occurred to me that if you were able to do this for several model species, you'd have a fantastic tool,” he says.Drosophila, yeast, and mouse databases. They called it the Gene Ontology, and it is now a major bioinformatics project that covers over 1,800 species. Their original paper on the idea in Nature Genetics is one of the most highly cited in the field. “His achievement is not just to have built this system but also to have organised the consortium behind it. It is now one of the most used resources in all of biology,” says Valencia.But this insight initially met with little interest. “My first presentation, at ISMB in Greece in 1997, went down like a lead balloon,” he recalls. Eventually, he and three like-minded colleagues settled the matter in a bar at the Montreal ISMB in 1998. Together, they decided to set up a cross-species ontology to be used by the Drosophila genome in 1999. “The process turned me into a nervous wreck,” he jokes. He published his account of this roller-coaster experience in a short but entertaining book called Won for All: How the Drosophila Genome was Sequenced .He went on to collaborate with Gerry Rubin and Craig Venter in sequencing the “We're lucky to have such an inspirational figure in the community,” says Valencia. “This award has been well deserved for a number of years.”In the spring of 1997, Olga Troyanskaya  was workAnd so began the career of one of the most promising young researchers in bioinformatics, and a deserving winner of this year's Overton Prize. “She is one of these forces of nature, full of energy,” says Alfonso Valencia, chair of the ISCB awards committee.Troyanskaya herself talks with infectious enthusiasm about her work. “I've always been fascinated by the problems of biology,” she says. “I was just better at computer science and math than the wet lab research. And it seemed to me that there had to be a lot you could contribute with computer science that you couldn't do with experimental techniques alone.”From the University of Richmond, Troyanskaya moved to Stanford University to complete a PhD in biomedical informatics, under the supervision of Russ Altman, a bioinformatician, and David Botstein, a geneticist. “I wanted a setup that was close to real biological problems, and I got exactly that. I learned a great deal from both of them,” she says.In 2003, she moved to Princeton University as an assistant professor in the Department of Computer Science and the Lewis-Sigler Institute for Integrative Genomics. “I am fortunate that the computer science department appreciates the impact of computing in biology, and that I have many wonderful colleagues at both the department and in the Institute. I found several amazing collaborators, and this allowed me to begin a number of interesting projects.”One of the key problems she focuses on is making better use of the vast but unwieldy biological datasets in databases around the world. “So instead of focusing on one study, we can take the entirety of published data. That allows you to ask very specific questions in a data-driven way and to develop novel biological hypotheses,” she says.An important goal is to predict the function of genes or proteins. There have been many experimental approaches to determine what genes do and how they are controlled inside the cell. But this work tends to produce datasets that are large and noisy. Troyanskaya's approach is to develop new ways for extracting useful information from these datasets using techniques from computer science such as machine learning and data mining.“Computation by itself is often not enough to discover new biology but it can direct experimental work,” she says. And she has set up a wet lab to help test and validate the hypotheses that the computer science helps generate. In 2009, for example, she used this approach to identify 109 new proteins involved in mitochondrial biogenesis in yeast.This combined approach is one of the things that sets Troyanskaya apart, says Valencia. “She is one of the first to have come from the computational side and then moved into the experimental area to combine both,” he says.Understanding the function of individual genes is only a small part of a much bigger story. Many genes and proteins play multiple roles within a cell as parts of various networks of biological processes. Mapping out these networks and understanding how they work and interact with each other is yet another strand of her research. “She has made important contributions to systems biology,” says Valencia.The process of evaluating and validating computational predictions is an area requiring a broad collaboration to develop standards and methods that can be used to achieve a consensus about the results. To this end, Troyanskaya is collaborating with the curators of model organism databases and members of the Gene Ontology Consortium.Another problem that many researchers face is handling the data avalanches currently being generated. So Troyanskaya, in collaboration with Princeton colleagues Kai Li and Moses Charikar, is looking at ways to better search and visualise these huge datasets, something that is challenging because of high noise levels and the enormous volume of the data. “We are developing better ways to do this,” she says.PLoS Computational Biology and Bioinformatics. And she is involved in conferences: organizing, chairing tracks and program committees. “That is something that is very much appreciated,” says Valencia. “We are lucky to have her.”The awards committee was also impressed by Troyanskaya's service for the community. She is involved in the Society's two official journals, And there is surely more to come. Troyanskaya points to numerous questions that are driving her research forward. She wants to know, for example, how we can predict which genes are involved in kidney disease, to understand their function and their clinical role on a molecular level. She works on these questions in close collaboration with experimental researchers, such as Matthias Kretzler and his group from the University of Michigan, Ann Arbor. And she is passionate about finding ways to ask questions in a data-driven way, not just in a knowledge-driven way that relies on what we already know about biology. “These are the questions that I'm really interested in,” she says. “And we really haven't yet harnessed the full potential of our data collections.”http://www.iscb.org/ismbeccb2011. The conference will also feature parallel tracks for Proceedings of original research papers, Highlights of recently published papers, Special Sessions on emerging topics, Late Breaking Research of peer-reviewed abstract submissions, and Technology demonstrations and workshops presented by academic researchers and commercial vendors. The conference also displays a unique “Art and Science” exhibit of scientifically based artistic visual images and videos submitted, and offers a commercial and non-profit vendor exhibition.The full conference agenda and registration information for ISMB/ECCB 2011, where these ISCB award winners, along with four other distinguished Keynote lecturers, can be found on the conference Web site at http://www.iscb.org/iscb-awards.For a review of past ISCB award winners, please see


We evaluated the potentiality of plasma microRNAs  that were considered as novel biomarkers for acute coronary syndrome (ACS), including acute myocardial infarction (AMI) and unstable angina (UA).We initially identified plasma miR-122, -140-3p, -144, -720, -1225-3p, -2861, and -3149 as candidate miRNAs associated with AMI (≥2 fold and P < 0.05) by comparing expression differences of miRNAs among AMI, non-coronary heart disease (non-CHD) and stable angina (SA) groups, using miRNA microarrays (n = 8 independent arrays in each group). Those seven plasma miRNAs were further examined with qRT-PCR analyses in two replications including 111 and 428 patients separately, and the results demonstrated that plasma miR-122, -140-3p, -720, -2861, and -3149 were elevated in the ACS group vs. the non-ACS (non-CHD + SA) group (P < 0.01). The area under the receiver operating characteristic curve (AUC) of the five miRNAs for ACS classification was 0.838, 0.818, 0.865, 0.852, and 0.670, respectively , while the values reached 0.843 and 0.925 when simultaneously with miR-122 and -3149 or with miR-122, -2861, and -3149 together . In plasma of pigs after coronary ligation, miR-122 was increased from 180 min to 240 min and miR-3149 was augmented from 30 min to 240 min compared with the sham pigs .Plasma miR-122, -140-3p, -720, -2861, and -3149 were associated with and potentially novel biomarkers for ACS. Acute coronary syndrome (ACS), including acute myocardial infarction (AMI) and unstable angina (UA), is one of the most serious cardiovascular diseases and a major cause of mortality and morbidity worldwide. Timely revascularization therapy within 3 h after the onset of symptoms is recommended by guidelines to rescue the ischemic myocardium and, thereafter, could eventually reduce the mortality . TherefoRecently, microRNAs  have been demonstrated to be involved in the mechanism of ACS, including rupture of the atherosclerotic plaque , 5, platFor detailed methods, see Blood samples were obtained in the cardiac catheterization laboratory from 16 healthy individuals , 72 high-risk individuals (Highrisk), 81 stable angina (SA), 287 UA, and 115 AMI (including STEMI and NSTEMI) patients.The protocol of this study was approved by the Ethics Committee of Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, China. Written informed consent was obtained from each patient before enrolment.Thirty six Bama male minipigs (10 months old) were fed a high-cholesterol diet for 20 weeks. Then pigs were randomly assigned to Sham (n = 6) and AMI (n = 30) groups, and the AMI group was further divided into AMI with ventricular fibrillation  and AMI without VF (AMI-NVF) groups after the end of the operation. All pigs were anesthetized with a mixture of ketamine hydrochloride 700 mg and diazepam 30 mg intramuscularly and were continuously infused with the mixture (2 mg/kg per hour) intravenously. At the end of the experiment, the deeply anesthetized pig was killed by an injection of 15% KCl (1 ml/kg). Blood samples were collected before the operation and after 30, 60, 90, 120, 180, and 240 min of coronary ligation.All animals received humane care in compliance with the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health, USA. The animal experimental protocols and procedures were approved by the Care of Experimental Animals Committee of Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, China.Peripheral blood samples (5 mL) were collected in the cardiac catheterization laboratory, and were separated into plasma and cellular fractions within 2 h of collection.Total RNA was isolated from plasma with TRIzol LS Reagent  according to the manufacturer’s protocol.http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE66752). The candidate miRNAs that differentially expressed in AMI patients were further validated by miRNA stem loop quantitative real-time polymerase chain reaction (qRT-PCR) technology.miRNAs profile in human plasma of AMI, Health, Highrisk and SA groups was determined by the human miRNA array (Agilent miRNAs microarray Version 16.0). The microarray dataset is publicly available at GEO database .-ΔΔct method. All values of miRNAs are expressed as mean ± SE. Comparisons of parameters among ≥ 3 groups were analyzed by oneway ANOVA, followed by post hoc testing with Bonferroni correction. Independent-sample T test was used for 2 group comparisons. All P values were two-sided and differences were considered statistically significant at a value of P < 0.05. The receiver operating characteristic (ROC) curves were established for classification of ACS and non-ACS patients. All statistical calculations were performed by the SPSS 20.0.Patients’ baseline characteristics of different groups were compared by One-way ANOVA and Fisher’s exact test. For microarray analysis, the Mann-Whitney unpaired test was used for the three pairwise comparisons . For the data obtained by qRT-PCR, a widely used method to present relative expression of miRNA is the 2Expression profiles of 1205 human miRNAs were initially screened by microarray between the Health, Highrisk, SA, and AMI patients (n = 8 independent arrays in each group). No significant differences of clinical characteristics were found among the four groups .Quantification revealed that plasma miR-122, -140-3p, -144, -720, -1225-3p, -2861, and -3149 were up-regulated ≥2 fold in the AMI patients compared with the Health, Highrisk, and SA patients . These 7 miRNAs were selected as candidate miRNAs for further testing via qRT-PCR in human samples . BecauseThe seven candidate miRNAs were first tested with qRT-PCR in 111 human plasma samples, including 21 non-CHD, 30 SA, 30 UA, and 30 AMI patients, and no significant differences of clinical characteristics were found among the four groups . The resThe expression profiles of miR-122, -140-3p, -720, -2861, and -3149 were further examined with qRT-PCR in 428 additional subjects’ plasma samples, including 51 non-CHD, 43 SA, 257 UA, and 77 AMI patients, and no significant differences of clinical characteristics were found among the four groups . The resA total of 539 human plasma samples from the two replications, including 394 ACS patients and 145 non-ACS patients, were used as the resource of training data set for the construction of the miRNA panel for the classification of ACS and non-ACS patients. The expression of miR-122, -140-3p, -720, -2861, and -3149 was increased respectively by 20.7, 14.7, 49.3, 27.6, and 12.5 fold in the ACS group compared with the non-ACS group  . The accAnimal experiments were conducted to assess whether the effects of ACS on miRNAs plasma levels in humans were observed in a swine model of AMI. Evans blue and triphenyltetrazolium chloride (TTC) staining indicated the success of AMI model , Panel AIschemic heart disease accounts for approximately 30% of all deaths, causing 1.4 million deaths in the developed world and 5.7 million deaths in developing regions . While aIn this study, we reported that the levels of circulating miR-122, -140-3p, -720, -2861, and -3149 were significantly up-regulated in human ACS, in comparison with the non-ACS patients. The ability of the five miRNAs to distinguish the ACS patients from the non-ACS patients was shown by the ROC curves with the AUC of 0.838, 0.818, 0.865, 0.852, and 0.670, respectively , and themiR-122, regarded as liver-specific , plays cmiR-140-3p has been reported to be increased in CHD . miR-720In addition, our animal study found that the plasma levels of miR-122 and -3149 were not only higher in the AMI pigs in comparison with the sham pigs, but also higher in the AMI-VF pigs versus the AMI-NVF pigs. The upregulated miR-21 has been reported to promote atrial fibrillation (AF) in MI-induced heart failure in rats . miR-29 It should be noted that the levels of cTnI were not significantly different within 240 min after coronary ligation between the sham and AMI pigs with the success of AMI model documented clearly by the triphenyltetrazolium chloride (TTC) staining. Previous studies have demonstrated that the cTnI levels in blood of AMI patients began to rise after 4–8 h of myocardial injury . But in The potential limitations should be addressed. First, the sample size of current study is relatively small. Second, the generalizability of our findings is limited by the demographic differences and treatment characteristics of ACS patients. In addition, it would be ideal to have information on the time course for corresponding miRNA quantification in AMI patients. Therefore, further studies with larger sample size and patients with various CHD subtypes are needed to validate the diagnostic value of plasma miR-122 and -3149 for ACS.In conclusion, plasma miR-122 and miR-3149 are associated with ACS and might be potentially novel biomarkers for accelerating the diagnosis of ACS patients in emergency department.S1 Appendix(DOCX)Click here for additional data file.S1 Table(DOC)Click here for additional data file.S2 Table(DOC)Click here for additional data file.S3 Table(DOC)Click here for additional data file.S4 Table(DOC)Click here for additional data file.


Topological states of matter support quantised nondissipative responses and exotic quantum particles that cannot be accessed in common materials. The exceptional properties and application potential of topological materials have triggered a large-scale search for new realisations. Breaking away from the popular trend focusing almost exclusively on crystalline symmetries, we introduce the Shiba glass as a platform for amorphous topological quantum matter. This system consists of an ensemble of randomly distributed magnetic atoms on a superconducting surface. We show that subgap Yu–Shiba–Rusinov states on the magnetic moments form a topological superconducting phase at critical density despite a complete absence of spatial order. Experimental signatures of the amorphous topological state can be obtained by scanning tunnelling microscopy measurements probing the topological edge mode. Our discovery demonstrates the physical feasibility of amorphous topological quantum matter, presenting a concrete route to fabricating new topological systems from nontopological materials with random dopants. Apart from the intensive efforts to explore topological properties in crystalline materials, the study of such properties in amorphous materials has been rare. Here, Pöyhönen et al. predict a topological superconducting phase in an ensemble of randomly distributed magnetic atoms on a superconducting surface. While topological properties can be studied independently of local order, spatial symmetries play a central role in virtually all material realisations. This is emphasised by the fact that the theoretical search for new topological materials extensively employs band structures and reciprocal space. While topological states are generically robust to disorder which breaks spatial symmetries, this is typically established by treating the disorder as an additional feature in a well-defined clean system. Even topological Anderson insulators4, where moderate disorder actually gives rise to nontrivial topological properties, rely crucially on a specific band structure of the clean system. The concept of disorder, almost by definition, implies the existence of an underlying ordered reference state.Topological states are characterised by integer-valued invariants7. The wires have the advantage of potentially allowing topological computation, but to carry out this function they must be almost defect free, which illustrates a generic complication in top-down fabrication strategies. On the other hand, fabrication of topological matter with randomly distributed constituents, if possible, would avoid that obstacle and offer new opportunities. A recent discovery of a mechanical gyroscopic metamaterial8, albeit a purely classical system, suggests that also amorphous topological quantum matter could be achievable. By studying the properties of long-range hopping toy models, Agarwala and Shenoy9 pointed out the mathematical possibility of topological states with randomly localised states.The role of spatial symmetries in topological materials raises the question of how much spatial order is necessary for topological states to persist. In addition to the fundamental interest, possible realisations have far-reaching practical implications. The search for topological states has already moved beyond the elements found in nature to artificial man-made structures such as Majorana wiresIn this work, we propose the Shiba glass, depicted in Fig. 18 with supporting experimental evidence21. More recently, ferromagnetic 2D lattices have emerged as a promising platform for chiral superconductivity23 with a rich topological phase diagram25. Classical magnetic moments embedded in a gapped s-wave superconductor give rise to Yu–Shiba–Rusinov (YSR) subgap states26, localised subgap states which decay algebraically for distances smaller than the superconducting coherence length. In 2D superconductors, such as layered systems, thin films and surfaces, the decay of the wavefunctions from the deep-lying impurity has a functional form ξ and kF are the superconducting coherence length and the Fermi wave vector of the underlying bulk. The Shiba glass results from a hybridisation of randomly distributed YSR states. To model the system, we consider deep-lying YSR states with energies ε0 located in the vicinity of the gap centre J is the magnetic coupling, S is the magnitude of the magnetic moment and ε0 ≈ Δ(1 − α). As outlined in the Methods section, the low-energy properties of the coupled impurity moments are modelled by a tight-binding Bogoliubov-de Gennes Hamiltonian24rn. The entries hmn, Δmn for arbitrary configuration of magnetic moments is lengthy and given in Supplementary Note The studied amorphous topological superconductor is comprised of randomly distributed magnetic moments on a superconducting surface with a Rashba spin-orbit coupling. The moments can arise from magnetic atoms, molecules or nanoparticles. Regular 1D structures of this type have been predicted to host Majorana statesxmn and ymn are components of rm − rn ≡ . The hopping elements are expressed in terms of the functionsJn and Hn are Bessel and Struve functions of order n. The Rashba spin-orbit coupling induces two helical Fermi surfaces with density of states λ = αR/(ħvF) is the dimensionless Rashba coupling and kF,vF the Fermi wavenumber and velocity in the absence of spin-orbit coupling. The Rashba coupling also slightly modifies the superconducting coherence length ij vanishes with vanishing Rashba coupling αR = 0. The low-energy Hamiltonian  oscillates as a function of the position as cos(2kFr). While for distances π/4 < kFr < 3π/4 the interaction is effectively ferromagnetic. Therefore, in the large part of the topological region BSZ would drive the system towards an out-of-plane polarisation.In our work, we have not addressed the question of the magnetic ordering, but rather show that a finite polarisation perpendicular to the plane gives rise to a topological phase. The nature of the ordering would likely depend sensitively on the specific physical realisation; however, there exist a number of mechanisms driving the system to a polarised state. For example, ignoring the modifications arising from superconductivity at large distancesd semiconductors with Rashba spin-orbit coupling are promising candidate systems. Another candidate system is the layered superconductor NbSe2 where 2d YSR states29 and their coupling have been observed30 recently. The most direct experimental probe is provided by STM measurement of the LDOS. As shown above, in the topological phase the Shiba glass system exhibits a significant concentration of the subgap LDOS at the sample boundaries, which can be directly observed by STM. This signal is clearly detectable at temperatures below the mobility gap scale which can be of the order of kBT = 0.1Δ − 0.3Δ as shown in Fig. The studied Shiba glass system could be realised by decorating an effective 2D or a layered 3D superconductor with magnetic atoms or molecules. Considering the requirement In summary, we introduced the Shiba glass as a platform for amorphous topological superconductivity and elucidated the general properties of such systems. Our results illustrate the physical feasibility of amorphous topological quantum matterials and provide a concrete prescription to experimentally realise and observe them. Our discovery motivates expanding the search for topological materials beyond crystalline systems and paves the way for fabricating topological matter from nontopological materials with random dopants.k-space, but there are various methods of computing it in real space as well32. A comparison shows that these methods are generally of similar computational efficiency and yield the same values for the topological invariant.To find the topological phase diagram, we need to evaluate topological invariants in real space. The relevant topological index for 2D systems with broken time-reversal symmetry is the Chern number. This is generally obtained in 32 proceeds by defining the coupling matrices Cα,α + 1, with elementsR is the position operator, qα = π for α = 0, …, 3, and where ψm are the eigenfunctions of the system with periodic boundary conditions. By use of these matrices, the Chern number is then obtained through the equationλm being the complex eigenvalues of the matrix C01C12C23C30.The real-space Chern number method of ref. Data sharing not applicable to this article as no datasets were generated or analysed during the current study.Supplementary InformationPeer Review File


Diabetic nephropathy (DN) is the most serious microvascular complication of diabetes and the largest single cause of end-stage renal disease (ESRD) in many developed countries. DN is also associated with an increased cardiovascular mortality. It occurs as a result of interaction between both genetic and environmental factors. Hyperglycemia, hypertension, and genetic predisposition are the major risk factors. However, the exact mechanisms of DN are unclear. Despite the benefits derived from strict control of glucose and blood pressure, as well as inhibition of renin-angiotensin-aldosterone system, many patients continue to enter into ESRD. Thus, there is urgent need for improving mechanistic understanding of DN and then developing new and effective therapeutic approaches to delay the progression of DN. This review focuses on recent progress and future perspective about mechanistic insight and management of DN. Some preclinical relevant studies are highlighted and new perspectives of traditional Chinese medicine (TCM) for delaying DN progression are discussed in detail. These findings strengthen the therapeutic rationale for TCM in the treatment of DN and also provide new insights into the development of novel drugs for the prevention of DN. However, feasibility and safety of these therapeutic approaches and the clinical applicability of TCM in human DN need to be further investigated. Diabetic nephropathy (DN) is a major complication of diabetes and the largest single cause of end-stage renal disease (ESRD) in many developed countries. DN is also associated with an increased cardiovascular mortality. Immunologic derangement is the cornerstone of pathogenesis of CKD . Of courDN is one of the most frequent and severe complications of diabetes mellitus and a worldwide public health problem affecting millions of people. Currently, based on the criteria of the kidney disease, Improving Global Outcomes (KDIGO) Clinical Practice Guideline, DN was diagnosed by renal biopsy or medical history . DN is tThe prevalence of DN has been dramatically increased in recent decades; especially in China, the adjusted prevalence of diabetes mellitus and prediabetes among Chinese adults is 9.7% and 15.5%, respectively , 19. In DN occurs as a result of interaction between both genetic and environmental factors. Hyperglycemia, hypertension, and genetic predisposition are the major risk factors. The aggravating hyperglycemia is also a high risk factor for the development of microvascular complications . In othePodocyte damage occurs at a relatively early stage in CKD . PodocytThe management of diabetes mellitus hinges on “five carriages,” which are health education, diet, exercise, weight control, and drug treatment, respectively, of which the former four therapies are described as nonpharmacological measures. And the pharmacological agents currently approved for treatment of diabetes mellitus include sulfonylureas, metformin, acarbose, and insulin . Nonpharβ-cells function of pancreatic island [β-cells of pancreatic island [There is no doubt that glycemic control plays the most momentous role in management of DN; keeping blood glucose within normal limits is a foundation and prerequisite for the treatment of DN . Severalc island , 38. Henc island . Moreovec island . Sulfonyc island . In addic island –44. TakeProteinuria is considered as a hallmark and sensitive marker of DN, which is mainly caused by the severe podocyte injury . But whaAt the same time, many patients of DN still show merging symptoms, such as dyslipidemia and hypertension. Poorly controlled blood glucose levels in patients with DN are associated with dyslipidemia which constitutes an additional risk factor. Dyslipidemia consists of elevated levels of low density lipoprotein cholesterol and low levels of high-density lipoprotein cholesterol and increased levels of apolipoproteins B . In viewCutting-edge evidence shows that there are certain trends of the future therapies for DN, including comprehensive exploration of existing medicines, application of molecular biology discoveries, progress in stem cell therapy, and the applicability of traditional Chinese medicine (TCM) in the treatment of DN.As mentioned previously, The RAAS pathway is very important in the progression of DN. Several studies indicate that the combination of spironolactone with an ACEI or ARB could significantly mitigate proteinuria compared with the placebo , 54. Furβ-cell receptors, stimulate insulin release, and improve glycemic control and then play a protecting role in management of DN [κB is a ubiquitous nuclear transcription factor that regulates expression of a large number of genes that are critical for the regulation of inflammation [Studies performed have discovered several key signaling pathways in succession, such as endoplasmic reticulum (ER) stress, which already have become a therapeutic target of much concern for DN . Besidesnt of DN . Moleculammation . The conammation .Stem cells that are identified nearly half a century ago exhibited great potential for the repair of damaged tissues and organs and provided new hope for a means to change the track of DN . Multiplα3β1 integrin upregulation and astragaloside IV could inhibit podocyte apoptosis by downregulation of PERK-ATF4-CHOP pathway [TCM has long histories in China and has emerged and influenced hundreds of thousands of people. Historically, conventional medicines, such as ACEI or ARB, are not able to totally prevent the development of DN . Thus, t pathway , 72. Fur pathway . Yiqi Ya pathway . These fIt is a long time that the relationship between hypertension and DN has been the subject of controversy, and after diabetes mellitus, hypertension is the second most commonly reported etiology of ESRD in the United States Renal Data System, and hypertension treatment targets in patients with DN remain important clinical concerns . Recent Despite the benefits derived from strict control of glucose and blood pressure, many patients continue to enter into ESRD. Thus, there is urgent need for improving mechanistic understanding of DN and then developing new and effective therapeutic approaches to delay the progression of DN. Many studies strengthen the therapeutic rationale for TCM in the treatment of DN. However, feasibility and safety of these therapeutic approaches and the clinical applicability of TCM in human DN need to be further investigated.


We propose a novel mechanism of enhancement of turbulence by energetic-particle-driven geodesic acoustic modes (EGAMs). The dynamics of drift-wave-type turbulence in the phase space is investigated by wave-kinetic equation. Spatially inhomogeneous turbulence in the presence of a transport barrier is considered. We discovered that trapping of turbulence clumps by the EGAMs is the key parameter that determines either suppress or enhance turbulence. In regions where turbulence is unstable, EGAM suppresses the turbulence. In contrast, in the stable region, EGAM traps clumps of turbulence and carries them across the transport barrier, so that the turbulence can be enhanced. The turbulence trapped by EGAMs can propagate independent of the gradients of density and temperature, which leads to non-Fickian transport. Hence, there appear a new global characteristic velocity, the phase velocity of GAMs, for turbulence dynamics, in addition to the local group velocity and that of the turbulence spreading. With these effect, EGAMs can deteriorate transport barriers and affect turbulence substantially. This manuscript provides a basis to consider whether a coherent wave breaks or strengthen transport barriers. The coexistence of global-coherent Alfven wave and micro-turbulence has been observed at the surface of the sun, which could link with the coronal heating problem3. Turbulence in planetary atmospheres generates zonal flows2, such as jet stream on the earth, stripes of Jupiter, super rotation on Venus, and tachocline on the sun4. In magnetically confined plasmas, geodesic acoustic modes (GAMs), which are oscillatory zonal flows, have attracted much attention as coherent waves, because GAMs are expected to suppress turbulence by their velocity shear1. Actually, the suppression of turbulence and transport have been reported in turbulence simulations5. Experimental study has shown that the transition to high confinement state is accompanied by GAMs6. However, recently, the enhancement of turbulence by GAMs has been observed in first principle simulations, with the subsequent destruction of a transport barrier9. In this way, GAMs can either mitigate or enhance the turbulence. This dual effect of the GAMs on turbulence requires theoretical investigation.Problems including interactions between micro-turbulence and coherent waves are ubiquitous in a variety of systems10 but also by energetic particles (EPs), which are called EGAM14. Turbulence driven GAMs suppress turbulence, which can be understood in terms of energy conservation . In contrast, the impacts of EGAMs on turbulence is not clear, in particular because EGAMs obtain their energy from EPs, not from turbulence. Actually, EGAMs have been reported to enhance turbulence in spite of the fact that EGAMs have velocity shears8. In order to clarify the impact of finite-frequency zonal flows on turbulence, we focus on EGAMs. The phase-space dynamics results in trapping of turbulence wave-packets by EGAMs15. We found that the trapped turbulence wave-packets leak across the transport barrier. As a result, turbulence is enhanced by EGAMs in the stable region, while turbulence suppression is obtained in the unstable region. We discovered that trapping of turbulence clumps by the EGAMs is the key parameter that determines either suppress or enhance turbulence. The propagation of the turbulence is ballistic, with the phase velocity of the EGAM. Thus, the turbulence propagation is in some sense independent from the background profiles such as the gradients of density and temperature. Thus, turbulence carried by the EGAMs shows non-Fickian transport properties. The propagation of trapped turbulence is different from processes such as turbulence spreading18, avalanches21 and others23. Hence, there appear a new global characteristic velocity, the phase velocity of GAMs, for turbulence dynamics, in addition to the local group velocity and that of the turbulence spreading.We investigate the phase-space dynamics of spatially inhomogeneous turbulence in the presence of GAMs. GAMs are driven not only by turbulencex-direction. We consider a situation where the turbulence unstable region faces the stable region, and a transport barrier is localized at the boundary, as shown in Fig. qy ≈ 011 and another has a steep poloidal structure24. In this study, we focus on the branch with qy ≈ 0, which is unaffected by the doppler-shift due to the mean sheared flow. This branch is driven by the resonance with the toroidally passing EPs, where we neglect the effect of the magnetically trapped EPs. In such a situation, the turbulence is governed by the wave-kinetic equation25,Nk and ωk are the normalized action and frequency of the turbulence, respectively25. The linear growth rate and the nonlinear decorrelation rate of turbulence are denoted by γL, and Δω, respectively. Here time and space are normalized by ρs, where Vd is the diamagnetic drift velocity, and ρs is the ion gyro-radius measured by the sound velocity. It is noted that the typical frequency of the turbulence satisfies the relation, ωEP is the transit frequency of EPs. Thus, we assume that the resonance of the EPs with the turbulence is weaker than that with EGAMs. We treat γL as an independent parameter on EPs. We consider drift-wave-type turbulence, so Nk is given as ωk iskx, ky are the turbulence wavenumbers. The turbulence frequency ωk includes the doppler shift due to We consider the dynamics of spatially inhomogeneous turbulence with a transport barrier in the presence of EGAMs. We focus on the impact of EGAM on turbulence. As the first step, in order to simplify the problem, we neglect the direct effect of EPs on turbulence, as shown in Fig. 26. Here, we compare the assumptions and results between these previous studies26 and the present work. In these studies, an analytic solution for a nonlinear wave pattern for zonal flows was obtained by considering the interaction between turbulence driven zonal flows and turbulence. The following assumptions were used; spatially homogeneous turbulence with a marginal stability condition was assumed  are shown in Fig. VG is the amplitude of the E × B velocity of the EGAM. The kx-spectrum of the turbulence in the unstable region x < 20 has positive-negative symmetry with respect to kx. In the stable region, the kx-spectrum becomes asymmetric, and the turbulence exists only for kx < 0, where the group velocity of the turbulence is positive. This outflow of the turbulence into the stable region can be understood by considering the motion of the quasi-particles (QPs). The equation of motion for the QPs is governed by Eq. , turbulence trapped by the EGAM forms islands in the phase space. As seen from the time evolution of the turbulence intensity in Fig. ωk becomes an adiabatic constant of motion. From Eq.  in space, with the typical scales, VG varies. The island in the case of n is an integer, and kx-spectra of ensemble averaged Nk at X-point, VG, the intensity of the turbulence increases inside the island We describe two mechanisms by which the turbulence propagates in the stable region; one occurs without the EGAM (transit loss), and another is due to the turbulence trapping by the EGAM (trapped loss). We investigate conditions for trapped loss to be dominant. Figure For our parameters, this condition is When both conditions, Eqs  and 10)10), are x = 0 (the unstable region) on the EGAM amplitude. The intensity N is calculated from the ensemble average, and the error bar is estimated from the standard deviation. The intensity N, which is calculated from the whole region in kx-space, decreases with VG, whose amplitude dependence is fitted to x = 27, is shown in Fig. VG, which clearly shows that the EGAM can enhance turbulence. The total N in the stable region scales as α is a constant parameter, VG and the spectral width increases with We study the dependence of the turbulence intensity 27. In the case of the gyrokinetic simulations by Zarzoso and others8, the shear layer is much narrower than the EGAM wavelength, The present mechanism of turbulence enhancement is discussed below in the contexts of experiments, and first-principle simulations. In the LHD, large amplitude EGAMs are observed with amplitude 19. The turbulence intensity is determined by that of the trapped turbulence Itrap, while the theoretical expression of the turbulence intensity of the avalanche has not been not reported. The radial flux of the turbulence clump by the trapped turbulence by the EGAM is qx is small, and/or the amplitude is large. The propagation distance of the trapped turbulence is Finally, the propagation property of the trapped turbulence is compared with the other processes. The propagation characteristics are summarized in Table In conclusion, two novel effects of the EGAM on turbulence are proposed by studying the phase-space dynamics of turbulence. Spatially inhomogeneous turbulence in the presence of transport barrier is considered. In the turbulence unstable region, the shear of the EGAM suppresses the turbulence. On the other hand, the turbulence trapped by EGAM leaks into the stable region across the transport barrier. Hence, turbulence is enhanced by the EGAM. The condition for the wave-trapping effect to be dominant is obtained, which is consistent with the simulations. The trapped turbulence by the EGAM can propagate independently from the gradients of density and temperature, which leads to the non-Fickian transport. Hence, there appear a new global characteristic velocity for turbulence dynamics, in addition to the local group velocity and that of the turbulence spreading. This manuscript provides a basis to consider whether a coherent wave breaks or strengthen transport barriers.ky-spectrum is assumed for simplicity26, and the time evolution of Nk is calculated in the phase space of A monochromatic VG is a given parameter, since the EGAM survives regardless of the presence of turbulence. We implicitly assume that the scale length of the EP-density is much longer than the wavelength of the EGAM. The calculation region is Nk with respect to x and kx are set to be zero at the boundaries. The simulations are performed with the set of the parameters;Nk numerically.Here,