Variable Selection for Support Vector Machines in Moderately High Dimensions.
The support vector machine (SVM) is a powerful binary classification tool with high accuracy and great flexibility. It has achieved great success, but its performance can be seriously impaired if many redundant covariates are included. Some efforts have been devoted to studying variable selection for SVMs, but asymptotic properties, such as variable selection consistency, are largely unknown when the number of predictors diverges to infinity. In this work, we establish a unified theory for a general class of nonconvex penalized SVMs. We first prove that in ultra-high dimensions, there exists one local minimizer to the objective function of nonconvex penalized SVMs possessing the desired oracle property. We further address the problem of nonunique local minimizers by showing that the local linear approximation algorithm is guaranteed to converge to the oracle estimator even in the ultra-high dimensional setting if an appropriate initial estimator is available. This condition on initial estimator is verified to be automatically valid as long as the dimensions are moderately high. Numerical examples provide supportive evidence.


Is there a relationship between resistin levels and left ventricular end-diastolic pressure?
Resistin, a cysteine-rich peptide, is associated with atherosclerosis and diabetes. Resistin levels increase corresponding to coronary artery disease (CAD) and heart failure severity. Since resistin level tends to elevate with symptomatic heart failure, it is expected to be associated with left ventricular end-diastolic pressure (LVEDP). However, there is no relevant literature on the relationship between resistin levels and LVEDP. We aimed to evaluate the association between resistin levels and LVEDP, severity of CAD, carotid intima-media thickness (CIMT), and echocardiographic diastolic dysfunction parameters. For this study, 128 euvolemic patients with creatinine clearance >50 mg/dL and without acute coronary syndrome, who had typical chest pain or were stress test positive, were enrolled. Resistin level was measured by Enzyme-linked immunosorbent assays (ELISA) method. Severe CAD is defined as ≥50% stenosis in one of the major coronary arteries. LVEDP was measured during left heart catheterization. After coronary angiography, 60 patients (46.9%) had severe CAD. The mean LVEDPs were similar for patients with and without severe CAD (p=0.480). The resistin levels did not differ between the groups (p=0.154). The resistin levels did not correlate with LVEDP (r=-0.045, p=0.627), ejection fraction (EF; r=0.110, p=0.228), the Gensini score (r=-0.091, p=0.328), and CIMT (r=0.082, p=0.457). No significant correlation was found between the echocardiographic diastolic dysfunction parameters and resistin levels. There was no significant correlation between resistin level and LVEDP, CAD severity, echocardiographic diastolic dysfunction parameters, and CIMT. Further studies are warranted to determine the efficacy of resistin in clinical use.


Mold and endotoxin levels in the aftermath of Hurricane Katrina: a pilot project of homes in New Orleans undergoing renovation.
After Hurricane Katrina, many New Orleans homes remained flooded for weeks, promoting heavy microbial growth. A small demonstration project was conducted November 2005-January 2006 aiming to recommend safe remediation techniques and safe levels of worker protection, and to characterize airborne mold and endotoxin throughout cleanup. Three houses with floodwater lines between 0.3 and 2 m underwent intervention, including disposal of damaged furnishings and drywall, cleaning surfaces, drying remaining structure, and treatment with a biostatic agent. We measured indoor and outdoor bioaerosols before, during, and after intervention. Samples were analyzed for fungi [culture, spore analysis, polymerase chain reaction (PCR)] and endotoxin. In one house, realtime particle counts were also assessed, and respirator-efficiency testing was performed to establish workplace protection factors (WPF). At baseline, culturable mold ranged from 22,000 to 515,000 colony-forming units/m3, spore counts ranged from 82,000 to 630,000 spores/m3, and endotoxin ranged from 17 to 139 endotoxin units/m3. Culture, spore analysis, and PCR indicated that Penicillium, Aspergillus, and Paecilomyces predominated. After intervention, levels of mold and endotoxin were generally lower (sometimes, orders of magnitude). The average WPF against fungal spores for elastomeric respirators was higher than for the N95 respirators. During baseline and intervention, mold and endotoxin levels were similar to those found in agricultural environments. We strongly recommend that those entering, cleaning, and repairing flood-damaged homes wear respirators at least as protective as elastomeric respirators. Recommendations based on this demonstration will benefit those involved in the current cleanup activities and will inform efforts to respond to future disasters.


Association of HLA DRB1 alleles with juvenile idiopathic arthritis in Mexicans.
The aim of the study was to investigate association between HLA class II alleles and juvenile idiopathic arthritis (JIA) in Mexican patients. We typed 120 patients with JIA and 99 healthy controls for HLA class II alleles were performed by PCR-SSO. Differences between the whole group of JIA and its subtypes and controls were calculated by using the Xi2; p-values were corrected (pc) with Bonferroni's test. The alleles HLA-DRB1*01 (pc= 0.00083) and HLA-DRB1*04 (pc=0.0049) were strongly associated with systemic JIA, while HLA-DRB1*11 and HLA-DRB1*14 were found to have decreased frequencies in the patients with systemic JIA compared to the controls. Two alleles were found to have increased frequencies with JIA oligoarthritis subgroup, HLA-DRB1*11 (p=0.01, pc=NS) and HLA-DRB1*13 (p=0.01, pc=NS). The HLA-DRB1*04 was found increased frequencies with susceptibility for RF negative and RF positive polyarthritis JIA subgroups (p correction resulted in loss of significance). In contrast two alleles HLA-DRB1*07 and HLA-DRB1*14 were found decreased frequencies only patients RF positive polyarthritis JIA subgroup compared to the controls (pc=NS). The profile of HLA-DRB1 alleles associations in Mexican with JIA were somewhat distinct from association typically found in Caucasians.


Temporal regulation of the expression of syncytin (HERV-W), maternally imprinted PEG10, and SGCE in human placenta.
Maternally imprinted PEG10 and SGCE, separated by approximately 2.15 Mb from Syncytin (HERV-W) gene at 7q21.3, are implicated in choriocarcinoma and Silver-Russell syndrome. Here we have analyzed the temporal regulation of mRNA expression of these genes in placenta and demonstrate that Syncytin gene activation is highest in term placenta, PEG10, downregulated at early hypoxic phase, and highly activated at 11-12 wk of gestation. In contrast, transcription from SGCE remained unchanged throughout pregnancy, suggesting two neighboring imprinted genes are differentially regulated at very early pregnancy. Additionally, accumulation of two major species of mRNA (8 kb and 3.1 kb) encoded by HERV-W in placenta is regulated: 3.1 kb mRNA level remained unchanged throughout pregnancy, whereas the production of 8 kb species was highest in term placenta. Western blot and immunohistochemical staining of placental tissues with monoclonal antibodies revealed a marked reduction of syncytin glycoprotein synthesis in late pregnancy. Therefore, the relative levels of 3.1 kb and 8 kb mRNAs in trophoblasts could regulate syncytin protein synthesis, possibly by competition of the two mRNA species for translational apparatus.


HBV-related HCC, clinical issues and therapy.
Every year, more than 600 thousand persons will develop hepatocellular carcinoma (HCC) worldwide. Sixty percent of all HCCs in Asia and Africa are related to chronic infection with the hepatitis B virus (HBV) compared to only 20% of HCC cases in Europe, Japan and USA. Surveillance of chronically infected patients improves treatment of HCC, since it helps detecting tumors amenable to radical therapies. In cirrhotic patients, HCC can be confidently diagnosed by contrast imaging techniques, albeit the accuracy of radiological diagnosis is largely influenced by tumor size. Hepatic resection and liver transplantation are the first therapeutic options for patients within Milan criteria. The long-term outcome of resection, however, is affected by high risk of tumor recurrence, whereas liver transplantation has a negligible risk of both HBV and HCC recurrence-related anticipated mortality. The clinical benefits of loco-regional ablative techniques are evidence based for patients with less than 3 cm tumors only, whereas the standard of care for intermediate tumors, chemoembolization, needs to be validated in view of new technical improvements. Tertiary prevention of HCC in patients with established HBV infection is doubtful, mainly due to methodological weaknesses of studies based on interferon and nucleos(t)ide analogues therapy.


Regulation of the sacPA operon of Bacillus subtilis: identification of phosphotransferase system components involved in SacT activity.
The sacT gene which controls the sacPA operon of Bacillus subtilis encodes a polypeptide homologous to the B. subtilis SacY and the Escherichia coli BglG antiterminators. Expression of the sacT gene is shown to be constitutive. The DNA sequence upstream from sacP contains a palindromic sequence which functions as a transcriptional terminator. We have previously proposed that SacT acts as a transcriptional antiterminator, allowing transcription of the sacPA operon. In strains containing mutations inactivating ptsH or ptsI, the expression of sacPA and sacB is constitutive. In this work, we show that this constitutivity is due to a fully active SacY antiterminator. In the wild-type sacT+ strain or in the sacT30 mutant, SacT requires both enzyme I and HPr of the phosphotransferase system (PTS) for antitermination. It appears that the PTS exerts different effects on the sacB gene and the sacPA operon. The general proteins of the PTS are not required for the activity of SacY while they are necessary for SacT activity.


Patient, Provider, and Practice Characteristics Associated With Sacubitril/Valsartan Use in the United States.
Background Current guidelines recommend sacubitril/valsartan for patients with heart failure with reduced ejection fraction, but the rate of adoption in the United States has been slow. Methods and Results Using data from CHAMP-HF (Change the Management of Patients With Heart Failure), we described current sacubitril/valsartan use and identified patient, provider, and practice characteristics associated with its use. We considered patients to be on sacubitril/valsartan if they were prescribed it before enrollment or initiated on it at the baseline visit. We excluded patients with a contraindication to sacubitril/valsartan and practices with <10 patients enrolled. Of 4216 patients from 121 sites, 616 (15%) were prescribed sacubitril/valsartan, 2506 (59%) an angiotensin-converting enzyme (ACE) inhibitor/angiotensin receptor blocker (ARB), and 1094 (26%) neither. Patients prescribed sacubitril/valsartan were younger (63 years versus 66 years ACE inhibitor/ARB versus 69 years neither, P<0.001), less likely to have chronic kidney disease (15% versus 17% ACE inhibitor/ARB versus 30% neither, P<0.001), more likely to have cardiac resynchronization therapy (12% versus 7% ACE inhibitor/ARB versus 7% neither, P<0.001), and had lower ejection fraction (27% versus 30% ACE inhibitor/ARB versus 30% neither, P<0.001). Larger practices, based on number of cardiologists and advanced practice providers, were associated with the highest sacubitril/valsartan use. After multivariable adjustment, the number of advanced practice providers was associated with sacubitril/valsartan use (adjusted odds ratio,1.08; 95% CI, 1.03-1.14). Conclusions Despite current guideline recommendations, adoption of sacubitril/valsartan remains low. Provider and practice characteristics associated with sacubitril/valsartan use were related to general practice size and were not associated with practice characteristics specific for heart failure. Further research is needed to identify strategies for effective quality improvement interventions in chronic heart failure with reduced ejection fraction.


Model averaging, optimal inference, and habit formation.
Postulating that the brain performs approximate Bayesian inference generates principled and empirically testable models of neuronal function-the subject of much current interest in neuroscience and related disciplines. Current formulations address inference and learning under some assumed and particular model. In reality, organisms are often faced with an additional challenge-that of determining which model or models of their environment are the best for guiding behavior. Bayesian model averaging-which says that an agent should weight the predictions of different models according to their evidence-provides a principled way to solve this problem. Importantly, because model evidence is determined by both the accuracy and complexity of the model, optimal inference requires that these be traded off against one another. This means an agent's behavior should show an equivalent balance. We hypothesize that Bayesian model averaging plays an important role in cognition, given that it is both optimal and realizable within a plausible neuronal architecture. We outline model averaging and how it might be implemented, and then explore a number of implications for brain and behavior. In particular, we propose that model averaging can explain a number of apparently suboptimal phenomena within the framework of approximate (bounded) Bayesian inference, focusing particularly upon the relationship between goal-directed and habitual behavior.


A new mathematical model for avascular tumour growth.
The early development of solid tumours has been extensively studied, both experimentally via the multicellular spheroid assay, and theoretically using mathematical modelling. The vast majority of previous models apply specifically to multicell spheroids, which have a characteristic structure of a proliferating rim and a necrotic core, separated by a band of quiescent cells. Many previous models represent these as discrete layers, separated by moving boundaries. Here, the authors develop a new model, formulated in terms of continuum densities of proliferating, quiescent and necrotic cells, together with a generic nutrient/growth factor. The model is oriented towards an in vivo rather than in vitro setting, and crucially allows for nutrient supply from underlying tissue, which will arise in the two-dimensional setting of a tumour growing within an epithelium. In addition, the model involves a new representation of cell movement, which reflects contact inhibition of migration. Model solutions are able to reproduce the classic three layer structure familiar from multicellular spheroids, but also show that new behaviour can occur as a result of the nutrient supply from underlying tissue. The authors analyse these different solution types by approximate solution of the travelling wave equations, enabling a detailed classification of wave front solutions.


Assessing Discrepancies in Neurocognitive and Patient-Reported Measures of Brain Tumor Survivors.
To examine the association between performance-based neurocognitive and patient-reported cognitive function tests and identify characteristics that may explain observed discrepancies as a means to advance intervention development. 40 adults diagnosed with a primary brain tumor (PBT) (high-grade, n = 35) were recruited from two academic neuro-oncology clinics in North Carolina. Eligibility included a Mini-Mental State Examination score of 24 or greater, having completed cancer treatment, and having tumor stability. Participants completed performance-based neurocognitive and patient-reported cognitive function, demographic, and symptom assessment tests at one time point. Neurocognitive impairments included executive control, memory, and attention. Age, time since diagnosis, and tumor- or treatment-specific variables were not associated with neurocognitive or patient-reported cognitive function. Those reporting worse cognitive impairment tended also to report greater severity of PBT-specific and depressive symptoms. Patient-reported cognitive concerns warrant additional assessment for potential interventions to maintain function.


[Vancomycin-resistant enterococci (VRE). Recent results and trends in development of antibiotic resistance].
Enterococci (mainly E. faecalis, E. faecium) are important nosocomial pathogens predominantly affecting older and/or immunocompromised patients. The bacteria possess a broad spectrum of intrinsic and acquired antibiotic resistance properties. Among these, the transferrable glycopeptide resistance of the vanA and vanB genotypes in vancomycin-resistant enterococci (VRE; reservoir: E. faecium) as well as resistance to last resort antibiotics (e.g. linezolid and tigecycline) are of special concern. Enterococci (including VRE) are easily transferred in hospitals; however, colonizations are far more frequent than infections. Resistance frequencies for vancomycin in clinical E. faecium isolates have remained at a relatively constant level of 8-15% (but with local or regional variations) in recent years whereas frequencies for teicoplanin resistance have shown a slight decrease. Glycopeptide resistance trends correlate with a spread of hospital-associated E. faecium strains carrying the vanA and, with rising frequency in recent years, the vanB gene cluster, the latter being associated with teicoplanin susceptibility. This increased occurrence of vanB-positive E. faecium strains may be caused by an increased use of antibiotics selecting enterococci and VRE as well as due to methodological reasons (e.g. reduced EUCAST MIC-breakpoints for glycopeptides; increased use and sensitive performance of chromogenic VRE agars, increased use of molecular diagnostic assays).


Monocyte Based Correlates of Immune Activation and Viremia in HIV-Infected Long-Term Non-Progressors.
Background: Disease progression monitoring through CD4 counts alone can be inadequate in HIV infection as ongoing immune activation may result in Serious non-AIDS events (SNAEs). SNAEs involve monocyte activation driven chronic inflammation with significant sequelae observed even during HAART. Here, we attempted to delineate functional monocyte based signatures across stages of HIV disease progression. Methods: Participants spanning four cohorts were recruited-pre-ART (PA; <7 years of infection; n = 20), long-term non-progressors (LTNP; >7 years of infection, CD4 > 350 cells/μL, n = 20), individuals on therapy (ART; n = 18) and seronegative controls (SN; n = 15). Immunophenotyping of monocyte subsets and evaluation of expression of HIV-binding receptors-CD4 and CCR5, marker of immune activation- HLA-DR and M2 phenotype-mannose receptor (CD206) was followed by association of monocyte-specific parameters with conventional markers of disease progression such as absolute CD4 count, CD4/CD8 ratio, viral load, and T cell activation. Results: A significant expansion of intermediate monocytes (CD14++CD16+) with a concomitant decline in classical subset (CD14++CD16-) was observed in all infected cohorts compared to seronegative controls. In addition, an expansion of the non-classical subset (CD14+CD16++) was observed in long-term non-progressors. Dysregulation in monocyte subsets associated with CD4 count and CD4/CD8 ratio in PAs but not in LTNPs. We report for the first time that expression of CD206 is most prominent on intermediate monocytes which also have the highest expression of CD4, CCR5, and HLA-DR. Despite preserved CD4 counts, LTNPs had similar immune activation profiles to PAs, as evidenced by elevated HLA-DR expression across monocyte subsets. HLA-DR expression, similar to that in SNs, observed in the ART group indicated partial immune restoration within the monocyte compartment. Increased CD206 expression on monocytes together with frequency of activated CD4+ T lymphocytes (HLA-DR+CD38+) showed significant and positive association with viral load in LTNPs, but not PAs. Conclusion: Our results describe for the first time the presence of monocyte dysregulation involving increased activation in LTNPs, who, in spite of preserved CD4 counts, may remain susceptible to prolonged effects of systemic inflammation and highlight CD206, as a unique non-T correlate of viremia, in viremic non-progression.


Medicines in schools: a cross-sectional survey of children, parents, teachers and health professionals.
To describe how individual schools manage medicines and strategies for implementation of guidance, to determine the nature of problems perceived by children, parents, teachers and healthcare professionals (HCPs) in relation to medicines management in schools and to highlight differences between these perceptions. A cross-sectional survey study in which questionnaires were completed by children, their parents and carers, groups of HCPs and head teachers. There were 158 respondents to this survey. The management of medicines varies between schools and this reflects how policy guidance is interpreted and is revealed by the differences in experience described. Head teachers acknowledge that there is a lack of expertise about medicines among their staff and they rely on interpretation of and adherence to policy and procedure and compliance with training was used as a measure of good medicines management. There are inconsistencies in how information about medicines is communicated between the healthcare team, families and schools, and there is evidence that this communication is not always timely or effective. This results in problems with medicines at school. Parents emphasised the need for staff at school to understand their child's condition and their medicines. There are differences between how individual schools manage medicines and interpret policy guidance and discrepancies between the views of each stakeholder group. There is some evidence that medicines management does not always meet the needs of children and their families. Fewer than half of parents and HCPs are satisfied with how medicines are dealt with in schools.


Evaluation of peripheral arterial disease in prediabetes.
The prevalence of prediabetes in the world continues to increase. These patients have elevated the risk of atherosclerosis. The current study was designed to assess the prevalence of peripheral arterial disease (PAD) and its related risk factors in prediabetes patients. This was the case-control study in which 135 adults in three groups: Diabetes, prediabetes, and normal were studied. We evaluated the prevalence of PAD through the measurement of ankle-brachial index (ABI). All the patients were interviewed about demographic and medical data, including age, sex, disease duration, body mass index, hypertension (HTN), fasting blood glucose, hemoglobin A1C (HbA1C), lipid profile, and medication use. The prevalence of PAD in diabetes patients was higher than the normal group (8.5%vs. 0.0%) (P < 0.05), but the differences between prediabetes compared with diabetes and normal group were not significant. The mean level of ABI in normal, prediabetes, and diabetes group was (1.11 ± 0.11), (1.09 ± 0.12), and (1.05 ± 0.03) respectively (P < 0.1). There were marginally significant differences of ABI observed between the normal group and the diabetes group. The observed differences between groups in the ABI were significant after adjusting the effects of age and sex (P < 0.05). There was an association observed between ABI and HbA1C in diabetes patients (r = 0.249, P < 0.01) and a significant association seen between PAD and HTN in the prediabetes group (P < 0.01). Peripheral arterial disease is common in asymptomatic diabetes and prediabetes patients. Management of hypertensive prediabetes patients and early detection of PAD in this group as well as in asymptomatic patients is important.


Finite element modelling versus classic beam theory: comparing methods for stress estimation in a morphologically diverse sample of vertebrate long bones.
Classic beam theory is frequently used in biomechanics to model the stress behaviour of vertebrate long bones, particularly when creating intraspecific scaling models. Although methodologically straightforward, classic beam theory requires complex irregular bones to be approximated as slender beams, and the errors associated with simplifying complex organic structures to such an extent are unknown. Alternative approaches, such as finite element analysis (FEA), while much more time-consuming to perform, require no such assumptions. This study compares the results obtained using classic beam theory with those from FEA to quantify the beam theory errors and to provide recommendations about when a full FEA is essential for reasonable biomechanical predictions. High-resolution computed tomographic scans of eight vertebrate long bones were used to calculate diaphyseal stress owing to various loading regimes. Under compression, FEA values of minimum principal stress (σ(min)) were on average 142 per cent (±28% s.e.) larger than those predicted by beam theory, with deviation between the two models correlated to shaft curvature (two-tailed p = 0.03, r(2) = 0.56). Under bending, FEA values of maximum principal stress (σ(max)) and beam theory values differed on average by 12 per cent (±4% s.e.), with deviation between the models significantly correlated to cross-sectional asymmetry at midshaft (two-tailed p = 0.02, r(2) = 0.62). In torsion, assuming maximum stress values occurred at the location of minimum cortical thickness brought beam theory and FEA values closest in line, and in this case FEA values of τ(torsion) were on average 14 per cent (±5% s.e.) higher than beam theory. Therefore, FEA is the preferred modelling solution when estimates of absolute diaphyseal stress are required, although values calculated by beam theory for bending may be acceptable in some situations.


Association between bacterial vaginosis and Herpes simplex virus type-2 infection: implications for HIV acquisition studies.
Bacterial vaginosis (BV) and Herpes simplex virus type-2 (HSV-2) have been linked to an increased risk of HIV-1 acquisition. Recent research suggests an association between BV and HSV-2 acquisition, but the converse has not been studied. Here, we investigate whether an association exists between BV and HSV-2 infection We examined the determinants of BV occurrence in a cohort of female sex workers in Burkina Faso. Participants were followed every 3 months for diagnosis of genital infections and report of sexual behaviours. Factors associated with BV occurrence were assessed using generalised estimating equation models. We enrolled 273 women (mean age, 28 years) and conducted 812 follow-up visits (mean 2.93 visit per woman). Baseline seroprevalence of HIV-1, HSV-2 and recent syphilis were 31.5%, 70.1% and 0.4%, respectively, while baseline prevalence of BV, Trichomonas vaginalis (TV) and Candida albicans were 20.5%, 3.3% and 2.5%, respectively. In multivariable analysis, HSV-2 (relative risk (RR) = 1.73, 95% CI 1.12 to 2.65), HIV-1 (RR = 1.76, 95% CI 1.30 to 2.40), TV (RR = 1.5, 95% CI 1.0 to 2.3), and having > or = 3 sexual partners in the preceding week (RR = 2.2, 95% CI 1.1 to 4.6) were independently associated with BV, while hormonal contraception showed a protective effect (RR = 0.11, 95% CI 0.02 to 0.70). HSV-2 infection was associated with BV occurrence in this population. As HSV-2 is strongly linked to HIV-1 acquisition, studies assessing the cofactor effect of BV on HIV acquisition should control for the presence of HSV-2. Further studies are required to investigate the relative effect of asymptomatic HSV-2 shedding and/or genital ulcerations on BV occurrence.


Fatty acid ethyl esters in the blood as markers for ethanol intake.
To determine the clinical utility of fatty acid ethyl esters (FAEEs) in the blood as a short-term confirmatory marker for ethanol intake and a longer-term marker for ethanol intake after ethanol is no longer detectable. Single-center controlled clinical trial and a blinded comparison involving 48 blood samples that were positive, negative, or equivocal for blood ethanol. Seven healthy subjects (4 men and 3 women, aged 21 to 23 years) participated in the clinical trial. Blood samples from participants for the blinded comparison portion of the study were numbered from 1 to 48 and not identified by name. The 7 healthy subjects ingested a known amount of ethanol at a fixed rate. The concentration of FAEEs in the blood after ethanol intake was determined for a period of up to 24 hours. There was no intervention in the blinded comparison study. In the clinical trial, a pharmacokinetic analysis of FAEE concentration in the blood after ethanol intake was completed for 7 individuals whose blood ethanol level was elevated from 25 to 35 mmol/L. In the blinded comparison, the 48 blood samples that were positive, negative, or equivocal for blood ethanol were analyzed for FAEE concentration. In the clinical trial, the disappearance of FAEEs from the blood followed a decay curve that initially resembled the decay curve for blood ethanol. However, because of a very slow secondary elimination phase, the FAEEs were found to persist in the blood for at least 24 hours after ethanol intake was completed. In the blinded comparison, all 20 samples that were positive for ethanol were positive for FAEEs, 7 of 7 samples equivocal for ethanol were positive for FAEEs, and 21 of 21 negative samples for ethanol were negative for FAEEs. Serum concentration of FAEEs can serve as an excellent short-term confirmatory test for ethanol intake as well as a longer-term marker of ethanol ingestion. Measurement of FAEEs in the blood may be a more sensitive indicator of ethanol ingestion than the measurement of blood ethanol .


Will reduced radiation damage occur with very small crystals?
The primary event which occurs when an X-ray photon of energy less than 30 keV is absorbed in a protein crystal (or other organic material) is the production of a photoelectron with a similar energy to that of the absorbed photon. The electron then scatters inelastically off the surrounding material losing energy in the process. This reduction in energy takes place over track lengths of a few microm for 20 keV electrons. The vector distances between the initial and final positions of the photoelectrons are less than the track lengths owing to the non-linear tracks followed by the electrons. For crystals with smaller dimensions than the vector distances, a significant proportion of the energy could leave the crystal with the high-energy electrons. This could provide an advantage in terms of reduced radiation damage. In order to estimate the possible benefits, calculations of the electron tracks are given, initially using the continuous slowing-down approximation. A Monte Carlo approach is then used to provide more accurate values of the vector distance travelled by electrons inside a protein crystal. The calculations indicate that significant reductions in radiation damage could occur for crystals of a few microm in size. The benefits would be greater when operating at higher energies. In addition, a scheme for realising the possible benefits in a practical situation is described. This could then form the basis of trial experiments.


Molecular analyses of oral polio vaccine samples.
It has been suggested that the human immunodeficiency virus (HIV), and thus the acquired immunodeficiency syndrome (AIDS) it causes, was inadvertently introduced to humans by the use of an oral polio vaccine (OPV) during a vaccination campaign launched by the Wistar Institute, Philadelphia, PA, USA, in the Belgian Congo in 1958 and 1959. The "OPV/AIDS hypothesis" suggests that the OPV used in this campaign was produced in chimpanzee kidney epithelial cell cultures rather than in monkey kidney cell cultures, as stated by H. Koprowski and co-workers, who produced the OPV. If chimpanzee cells were indeed used, this would lend support to the OPV/AIDS hypothesis, since chimpanzees harbor a simian immunodeficiency virus, widely accepted to be the origin of HIV-1. We analyzed several early OPV pools and found no evidence for the presence of chimpanzee DNA; by contrast, monkey DNA is present.


Tacrolimus in pancreas transplantation: a multicenter analysis. Tacrolimus Pancreas Transplant Study Group.
This follow-up multicenter analysis is based on 362 pancreas allograft recipients at 14 institutions who were given tacrolimus between 1 May 1994 and 15 November 1995. Three groups were studied: (1) recipients given tacrolimus initially for induction and maintenance therapy (n = 250; 215 without, 35 with, a concurrent bone marrow transplant), (2) recipients who converted to tacrolimus for rescue or rejection therapy (n = 89), and (3) recipients who converted to tacrolimus for other reasons (n = 23). Of 215 recipients without a bone marrow transplant in the induction group, 166 (77%) underwent a simultaneous pancreas-kidney transplant (SPK), 29 (14%) a pancreas transplant alone (PTA), and 20 (9%) a pancreas after previous kidney transplant (PAK). Initial antibody therapy was given to 185 (86%) recipients. All 215 received tacrolimus and prednisone; 202 (94%) also received azathioprine (AZA) and 11 (5%) mycophenolate mofetil (MMF). The most common side effects of tacrolimus were neurotoxicity in 21%, nephrotoxicity in 21%, gastrointestinal (GI) toxicity in 13%, and diabetogenicity in 13% of these recipients. No recipient in this group developed new-onset insulin-dependent diabetes mellitus. Of 89 recipients in the rescue group, 71 (79%) had an SPK, 11 (13%) a PTA, and 7 (8%) a PAK. Before conversion, all had been on cyclosporine (CsA)-based immunosuppression; 74% of them had 2 or more rejection episodes previously. The most common side effects were nephrotoxicity in 27%, neurotoxicity in 26%, GI toxicity in 18%, and diabetogenicity in 8% of these recipients. No recipient in this group developed new-onset insulin-dependent diabetes mellitus. In the induction group patient survival at 1 yr was 98% for SPK, 79% for PTA, and 100% for PAK recipients. According to a matched-pair analysis, pancreas graft survival for SPK recipients at 1 yr was 88% with tacrolimus vs. 73% with CsA (p = 0.002); for PTA recipients, 68% vs. 70% (p > 0.35); and for PAK recipients, 85% vs. 65% (p = 0.13). Graft loss from rejection was not different with tacrolimus vs. CsA in all 3 pancreas recipient categories. At 1 yr, 17% of recipients had converted from tacrolimus to CsA for diabetogenicity, nephrotoxicity, or rejection; 23% had converted from AZA to MMF. The incidence of post-transplant lymphoma was < 2%. In the rescue group, patient survival rates at 1 yr were 96% for SPK, 100% for PTA, and 86% for PAK recipients (p < 0.08). Pancreas graft survival at 1 yr was 89% for SPK, 58% for PTA, and 69% for PAK recipients (p = 0.004). Graft loss from rejection was significantly lower for SPK vs. PTA or PAK recipients. At 1 yr, 20% of recipients had reconverted from tacrolimus to CsA for rejection, neurotoxicity, or nephrotoxicity; 19% had converted from AZA to MMF. There were no post-transplant lymphomas in the rescue group. This follow-up multicenter analysis shows that tacrolimus after pancreas transplantation is associated with high graft survival rates when used for induction and with high graft salvage rates when used for rescue therapy. The rate of graft loss from rejection is low in all 3 pancreas recipient categories. The overall incidence of new-onset insulin-dependent diabetes mellitus is < 1%, as is the incidence of post-transplant lymphoma. Converting from tacrolimus to CsA and, in patients on tacrolimus, from AZA to MMF, is safe; interchangeable use of drugs appears to be of immunologic benefit. To determine the best immunosuppressive regimen after pancreas transplantation, a prospective randomized study comparing tacrolimus and MMF vs. Neoral plus MMF is mandatory.


A cross-sectional survey of cannabis use by people with MS in Oregon and Southwest Washington.
Evidence supports that cannabinoids reduce self-reported spasticity and neuropathic pain in people with MS (PwMS), and legal access to cannabis for medical and recreational use continues to rise. However, there are limited data regarding patterns of cannabis use and perceived benefits of cannabis among PwMS in the US. This study describes the prevalence of cannabis use, routes of administration, perceived benefit of cannabis for MS, and characteristics associated with cannabis use and perception of benefit among a population of PwMS living in two states where cannabis is legal for both medical and recreational use. A survey about treatments used by PwMS, focusing on complementary and alternative medicine (CAM), was sent to PwMS living in Oregon and Southwest Washington. This survey included questions about current and past cannabis use, route of cannabis administration, and perceived benefits, as well as personal demographics. Of the 1188 returned surveys, 1000 had at least 75% complete survey responses and also completed the questions about current and past cannabis use. Thirty percent (n=303) of respondents reported currently using cannabis, 21% (n=210) used in the past but not currently, and 49% (n=487) had never used cannabis. Among current users, rates of use by smoking, vaping, topicals, tinctures and oils, or edibles were similar (35-46%), and most (59%) reported using multiple routes of administration. Most (64-78%, varying by route) current and past users reported cannabis being very or somewhat beneficial for their MS. The odds of current cannabis use were higher in PwMS who: 1) were younger (OR 2.24 [95% CI 1.39-3.61] for those age 18-40 compared with age >60]; 2) had lower household income (OR 3.94 [95% CI 2.55-6.09] with annual income <$25k compared with those with >$100k); 3) had secondary progressive MS (OR 1.77 [95% CI 1.07-2.92]); and 4) had more than minimal MS disability (OR 2.05 [95% CI 1.03-4.10] for those using a walker compared to those with none/minimal disability). The odds of perceiving cannabis as beneficial for MS were higher in: 1) younger individuals (OR 5.61 [95% CI 2.62-11.98] for those age 18-40 compared with age >60); 2) those with lower household income (OR 3.35 [95% CI 1.65-6.80] with annual income <$25k compared with those with >$100k), 3) those not currently using disease modifying therapies (OR 2.32 [95% CI 1.30-4.13]), and 4) those with the greatest disability (OR 17.96; [95% CI 2.00-161.22]). In this survey, 30% of PwMS reported currently using cannabis for their MS, mostly by multiple routes of administration, and most of these people report this being helpful for their MS. People who were younger, had lower household income, had progressive disease, and had more than minimal disability were more likely to use cannabis and report it was beneficial for their MS. People who were not using disease modifying therapies were also more likely to report benefit from cannabis use.


Are there consistent grazing indicators in Drylands? Testing plant functional types of various complexity in South Africa's Grassland and Savanna Biomes.
Despite our growing knowledge on plants' functional responses to grazing, there is no consensus if an optimum level of functional aggregation exists for detecting grazing effects in drylands. With a comparative approach we searched for plant functional types (PFTs) with a consistent response to grazing across two areas differing in climatic aridity, situated in South Africa's grassland and savanna biomes. We aggregated herbaceous species into PFTs, using hierarchical combinations of traits (from single- to three-trait PFTs). Traits relate to life history, growth form and leaf width. We first confirmed that soil and grazing gradients were largely independent from each other, and then searched in each biome for PFTs with a sensitive response to grazing, avoiding confounding with soil conditions. We found no response consistency, but biome-specific optimum aggregation levels. Three-trait PFTs (e.g. broad-leaved perennial grasses) and two-trait PFTs (e.g. perennial grasses) performed best as indicators of grazing effects in the semi-arid grassland and in the arid savanna biome, respectively. Some PFTs increased with grazing pressure in the grassland, but decreased in the savanna. We applied biome-specific grazing indicators to evaluate if differences in grazing management related to land tenure (communal versus freehold) had effects on vegetation. Tenure effects were small, which we mainly attributed to large variability in grazing pressure across farms. We conclude that the striking lack of generalizable PFT responses to grazing is due to a convergence of aridity and grazing effects, and unlikely to be overcome by more refined classification approaches. Hence, PFTs with an opposite response to grazing in the two biomes rather have a unimodal response along a gradient of additive forces of aridity and grazing. The study advocates for hierarchical trait combinations to identify localized indicator sets for grazing effects. Its methodological approach may also be useful for identifying ecological indicators in other ecosystems.


Innate immunity in ocular Chlamydia trachomatis infection: contribution of IL8 and CSF2 gene variants to risk of trachomatous scarring in Gambians.
Trachoma, a chronic keratoconjunctivitis caused by Chlamydia trachomatis, is the world's commonest infectious cause of blindness. Blindness is due to progressive scarring of the conjunctiva (trachomatous scarring) leading to in-turning of eyelashes (trichiasis) and corneal opacification. We evaluated the contribution of genetic variation across the chemokine and cytokine clusters in chromosomes 4q and 5q31 respectively to risk of scarring trachoma and trichiasis in a large case-control association study in a Gambian population. Linkage disequilibrium (LD) mapping was used to investigate risk effects across the 4q and 5q31 cytokine clusters in relation to the risk of scarring sequelae of ocular Ct infection. Disease association and epistatic effects were assessed in a population based study of 651 case-control pairs by conditional logistic regression (CLR) analyses. LD mapping suggested that genetic effects on risk within these regions mapped to the pro-inflammatory innate immune genes interleukin 8 (IL8) and granulocyte-macrophage colony stimulatory factor (CSF2) loci. The IL8-251 rare allele (IL8-251 TT) was associated with protection from scarring trachoma (OR = 0.29 p = 0.027). The intronic CSF2_27348 A allele in chromosome 5q31 was associated with dose dependent protection from trichiasis, with each copy of the allele reducing risk by 37% (p = 0.005). There was evidence of epistasis, with effects at IL8 and CSF2 loci interacting with those previously reported at the MMP9 locus, a gene acting downstream to IL8 and CSF2 in the inflammatory cascade. innate immune response SNP-haplotypes are linked to ocular Ct sequelae. This work illustrates the first example of epistatic effects of two genes on trachoma.


Mind and muscle: the cognitive-affective neuroscience of exercise.
There is growing basic-science interest in the mechanisms underpinning the positive effects of exercise on brain function and cognitive-affective performance. There is also increasing clinical evidence that exercise may prevent and treat various neuropsychiatric disorders. At the same time, there is growing awareness that athletic performance is mediated in crucial ways by central nervous system mechanisms. The relevant mechanisms in all these cases requires further exploration, but likely includes neurotrophic, neuroendocrine, and neurotransmitter systems, which in turn are crucial mediators of psychopathology and resilience. The hypothesis that Homo sapiens evolved as a specialist endurance runner provides an intriguing context against which to research the proximal mechanisms relevant to a cognitive-affective neuroscience of exercise.


Cytochrome cd1 nitrite reductase NirS is involved in anaerobic magnetite biomineralization in Magnetospirillum gryphiswaldense and requires NirN for proper d1 heme assembly.
The alphaproteobacterium Magnetospirillum gryphiswaldense synthesizes magnetosomes, which are membrane-enveloped crystals of magnetite. Here we show that nitrite reduction is involved in redox control during anaerobic biomineralization of the mixed-valence iron oxide magnetite. The cytochrome cd1-type nitrite reductase NirS shares conspicuous sequence similarity with NirN, which is also encoded within a larger nir cluster. Deletion of any one of these two nir genes resulted in impaired growth and smaller, fewer, and aberrantly shaped magnetite crystals during nitrate reduction. However, whereas nitrite reduction was completely abolished in the ΔnirS mutant, attenuated but significant nitrite reduction occurred in the ΔnirN mutant, indicating that only NirS is a nitrite reductase in M. gryphiswaldense. However, the ΔnirN mutant produced a different form of periplasmic d(1) heme that was not noncovalently bound to NirS, indicating that NirN is required for full reductase activity by maintaining a proper form of d1 heme for holo-cytochrome cd(1) assembly. In conclusion, we assign for the first time a physiological function to NirN and demonstrate that effective nitrite reduction is required for biomineralization of wild-type crystals, probably by contributing to oxidation of ferrous iron under oxygen-limited conditions.


Ethics in neonatology: a look over Europe.
Advances in perinatal medicine have dramatically improved neonatal survival. End-of-life decision making for newborns with adverse prognosis is an ethical challenge, the ethical issues are controversial and little evidence exists on attitudes and values in Europe. to assess the attitudes of the neonatal departments in perinatal clinical practice in the hospitals of European countries. a questionnaire was send to 55 NICUs from 19 European countries. Forty five (81.8%) NICUs were Level III. Religion was Christian in 90.7% and we observed that in north countries the religion is more influent on clinical decisions (p = 0.032). Gestational age was considered with no significant difference for clinical investment. North countries consider birth weight (p = 0.011) and birth weight plus gestational age (p = 0.024) important for clinical investment. In north countries ethical questions should not prevail when the decision is made (p = 0.049) and from an ethical point of view, there is no difference between withdraw a treatment and do not initiate the treatment (p = 0.029). More hospitals in south countries administer any analgesia (p = 0.007). When the resuscitation is not successful 96.2% provide comfort care. Our study reveals that cultural and religious differences influenced ethical attitudes in NICUs of the European countries.


Cerebral gray matter volume deficits after weight recovery from anorexia nervosa.
Structural changes have been observed in the brains of low-weight patients with anorexia nervosa (AN), including increased cerebrospinal fluid (CSF) volumes and decreased gray matter and white matter volumes. We hypothesized that subjects who are weight-recovered from AN would show elevated CSF volumes and reduced gray matter volumes compared with controls. We used magnetic resonance imaging to compare the brains of 12 subjects who are weight-recovered from AN (time since weight recovery, 1-23 years) with those of 18 healthy control subjects and 13 low-weight patients with AN. Axial, dual-echo scans of the whole brain were segmented into gray matter, white matter, and CSF compartments by means of a computerized volumetric approach. Brain measures were corrected for the significant effects of intracranial volume and age, based on regression analysis of a larger group of 30 healthy female controls. Tests showed that the weight-recovered group had significantly greater CSF volumes and smaller gray matter volumes than the control group. By comparison with low-weight patients, the weight-recovered subjects had significantly smaller CSF volumes and significantly larger gray matter and white matter volumes. In the weight-recovered group, neither the CSF elevations nor gray matter deficits were correlated with the length of time since weight recovery. The persistent gray matter volume deficits in subjects who are weight-recovered from AN suggest that there may be an irreversible component to the brain changes associated with the illness. The neuropathological features of this irreversible component have yet to be characterized.


SF3B1 haploinsufficiency leads to formation of ring sideroblasts in myelodysplastic syndromes.
Whole exome/genome sequencing has been fundamental in the identification of somatic mutations in the spliceosome machinery in myelodysplastic syndromes (MDSs) and other hematologic disorders. SF3B1, splicing factor 3b subunit 1 is mutated in 60%-80% of refractory anemia with ring sideroblasts (RARS) and RARS associated with thrombocytosis (RARS-T), 2 distinct subtypes of MDS and MDS/myeloproliferative neoplasms (MDSs/MPNs). An idiosyncratic feature of RARS/RARS-T is the presence of abnormal sideroblasts characterized by iron overload in the mitochondria, called RS. Based on the high frequency of mutations of SF3B1 in RARS/RARS-T, we investigated the consequences of SF3B1 alterations. Ultrastructurally, SF3B1 mutants showed altered iron distribution characterized by coarse iron deposits compared with wild-type RARS patients by transmission electron microscopy. SF3B1 knockdown experiments in K562 cells resulted in down-regulation of U2-type intron-splicing by RT-PCR. RNA-sequencing analysis of SF3B1 mutants showed differentially used genes relevant in MDS pathogenesis, such as ASXL1, CBL, EZH, and RUNX families. A SF3B pharmacologic inhibitor, meayamycin, induced the formation of RS in healthy BM cells. Further, BM aspirates of Sf3b1 heterozygous knockout mice showed RS by Prussian blue. In conclusion, we report the first experimental evidence of the association between SF3B1 and RS phenotype. Our data suggest that SF3B1 haploinsufficiency leads to RS formation.


The mechanical stability of ubiquitin is linkage dependent.
Ubiquitin chains are formed through the action of a set of enzymes that covalently link ubiquitin either through peptide bonds or through isopeptide bonds between their C terminus and any of four lysine residues. These naturally occurring polyproteins allow one to study the mechanical stability of a protein, when force is applied through different linkages. Here we used single-molecule force spectroscopy techniques to examine the mechanical stability of N-C-linked and Lys48-C-linked ubiquitin chains. We combined these experiments with steered molecular dynamics (SMD) simulations and found that the mechanical stability and unfolding pathway of ubiquitin strongly depend on the linkage through which the mechanical force is applied to the protein. Hence, a protein that is otherwise very stable may be easily unfolded by a relatively weak mechanical force applied through the right linkage. This may be a widespread mechanism in biological systems.


Coronary artery disease in pediatric cardiac transplant recipients receiving triple-drug immunosuppression.
Although triple-drug immunosuppression (cyclosporine, azathioprine, and prednisone) has reduced the incidence of acute rejection after pediatric cardiac transplantation, its effect on the development of coronary artery disease, which may be the major determinant of long-term survival in these patients, is not well defined. We reviewed 42 coronary angiograms obtained annually in 17 cardiac transplant recipients, aged 6 months to 18 years (mean, 12.4 years) at the time of transplantation, who had been maintained on triple-drug immunosuppression and had survived at least 1 year after transplantation. Each angiogram was reviewed for luminal irregularities or discrete stenoses, for the loss of third-order branching, and for the presence of myocardial bridging or calcification of vessels. Patient files were reviewed for donor and recipient age, sex, and ABO blood group, for postoperative episodes of rejection or cytomegalovirus infection, for hypertension, and for cholesterol and triglyceride values. No recipient in our series has died or undergone retransplantation because of coronary artery disease. Six of 17 (35%) patients have developed angiographically identifiable coronary artery abnormalities: four by the first year and two additional recipients by the second and third years, respectively, after transplantation. Development of coronary artery abnormalities approached a significant correlation when related to posttransplantation cytomegalovirus infection (p = 0.11) and older recipient age (p = 0.056) but not to any other factors studied, including episodes of rejection (p = 1.0). Angiographically identifiable coronary artery abnormalities can occur in pediatric recipients within the first year after cardiac transplantation in spite of a low incidence of acute rejection. Although the abnormalities may be mild initially, they can progress and require intervention.(ABSTRACT TRUNCATED AT 250 WORDS)


Population structure of the soft tick Ornithodoros maritimus and its associated infectious agents within a colony of its seabird host Larus michahellis.
The epidemiology of vector-borne zoonoses depends on the movement of both hosts and vectors, which can differ greatly in intensity across spatial scales. Because of their life history traits and small size, vector dispersal may be frequent, but limited in distance. However, little information is available on vector movement patterns at local spatial scales, and particularly for ticks, transmitting the greatest diversity of recognized infectious agents. To test the degree to which ticks can disperse and disseminate pathogens at local scales, we investigated the temporal dynamics and population structure of the soft tick Ornithodoros maritimus within a colony of its seabird host, the Yellow-legged gull Larus michahellis. Ticks were repeatedly sampled at a series of nests during the host breeding season. In half of the nests, ticks were collected (removal sampling), in the other half, ticks were counted and returned to the nest. A subsample of ticks was screened for known bacteria, viruses and parasites using a high throughput real-time PCR system to examine their distribution within the colony. The results indicate a temporal dynamic in the presence of tick life stages over the season, with the simultaneous appearance of juvenile ticks and hatched chicks, but no among-nest spatial structure in tick abundance. Removal sampling significantly reduced tick numbers, but only from the fourth visit onward. Seven bacterial isolates, one parasite species and one viral isolate were detected but no spatial structure in their presence within the colony was found. These results suggest weak isolation among nests and that tick dispersal is likely frequent enough to quickly recolonize locally-emptied patches and disseminate pathogens across the colony. Vector-mediated movements at local scales may therefore play a key role in pathogen emergence and needs to be considered in conjunction with host movements for predicting pathogen circulation and for establishing effective control strategies.


A Quasi-Experimental Evaluation of an Education Program for the Use of Electrical Stimulation Therapy to Heal Pressure Injuries.
Electrical stimulation (E-Stim) involves applying low levels of electrical current. Despite high-level recommendations for E-stim use in many pressure injury (PrI) best practice treatment guidelines, clinicians seldom use E-Stim. This quasi-experimental design study aimed to determine whether an educational program could improve health care providers' knowledge and attitudes regarding the use of E-Stim for treating PrIs in community-dwelling individuals with spinal cord injury living in 1 region of Ontario, Canada. An educational intervention based on a university-level continuing education program was developed as part of a multifaceted knowledge mobilization project. Health care providers (eg, nurses, physicians, and allied health professionals) from multiple agencies were invited to participate. The instructional series included 8 online modules on background theory and knowledge and a hands-on workshop that familiarized participants with the equipment necessary to deliver E-Stim. Knowledge (percentage of correct answers using a knowledge test developed by the research team) and attitudes (assessed using the E-Stim Attitude Survey in which items were scored using a 5-point Likert scale (where 0 indicated a negative attitude and 5 a positive attitude) were evaluated 3 times (pre-education, post-online, and post-workshop). Data were aggregated into unit-weighted averaged composites of 3 attitude subscales (resources, evidence-based practice, and education), which were compared before and after educational sessions using a linear mixed effect model. Among the 83 participants, a significant increase in knowledge scores was noted from pre-education (55.9%) to post-online (78.4%) and post-workshop (78.0%) (X² [2] = 89.34; P <.001). A significant increase in attitude scores was noted across time points (resources: X² [2] = 27.32, P <.0001; evidence-based practice: X² [2] = 38.93, P <.0001; and education: X² [2] = 92.88, P <.0001). For the evidence-based practice subscale, attitude increased significantly post-online (t[127] = 6.03, P <.0001). For the resources subscale, a significant increase was detected after post-workshop (t[113] = 5.23, P <.001]. Online education increased health care providers' knowledge about E-Stim; however, hands-on workshops were required to change certain attitudes about the use of E-Stim for wound healing. Further research is required to evaluate 1) whether a change in knowledge and attitude scores translates to a practice change for health care providers and 2) the potential importance of ongoing coaching and mentorship for a sustainable change in the clinical setting.


Twenty-Five New Viruses Associated with the Drosophilidae (Diptera).
Drosophila melanogaster is an important laboratory model for studies of antiviral immunity in invertebrates, and Drosophila species provide a valuable system to study virus host range and host switching. Here, we use metagenomic RNA sequencing of about 1600 adult flies to discover 25 new RNA viruses associated with six different drosophilid hosts in the wild. We also provide a comprehensive listing of viruses previously reported from the Drosophilidae. The new viruses include Iflaviruses, Rhabdoviruses, Nodaviruses, and Reoviruses, and members of unclassified lineages distantly related to Negeviruses, Sobemoviruses, Poleroviruses, Flaviviridae, and Tombusviridae. Among these are close relatives of Drosophila X virus and Flock House virus, which we find in association with wild Drosophila immigrans. These two viruses are widely used in experimental studies but have not been previously reported to naturally infect Drosophila. Although we detect no new DNA viruses, in D. immigrans and Drosophila obscura, we identify sequences very closely related to Armadillidium vulgare iridescent virus (Invertebrate iridescent virus 31), bringing the total number of DNA viruses found in the Drosophilidae to three.


Effects of far-infrared sauna bathing on recovery from strength and endurance training sessions in men.
This study investigated effects of far-infrared sauna (FIRS) bathing on recovery from strength training and endurance training sessions, but also possible differences between FIRS and traditional (TRAD) Finnish sauna bathing. Ten healthy physically active male volunteers had on various days either a 60 min hypertrophic strength training session (STS) or a 34-40 min maximal endurance training session (ETS), which was following by 30 min bathing in special FIRS sauna at temperature of 35-50°C and humidity of 25-35%. After the sauna, subjects sat for 30 min at room temperature (21°C and 25-30% humidity). In comparison, 30 min of TRAD took place at 35-50°C and in 60-70% humidity. Performance tests included maximal isometric bench press and leg press, counter movement jump (CMJ) and maximal oxygen uptake on a treadmill. After STS, there were decreases in maximal isometric bench press (p < 0.001), maximal isometric leg press (p < 0.001), CMJ (p < 0.001) and pH (p < 0.001), but increases in heart rate (p < 0.001) and lactate concentration (p < 0.001) as expected. During recovery there were no differences in any variables between FIRS and no sauna bathing (NO SAUNA). Maximal ETS increased oxygen uptake (p < 0.001), heart rate (p < 0.001), lactate concentration (p < 0.001) and decreased pH (p < 0.001) as expected. During recovery at 30 min, CMJ was significantly (p < 0.05) higher (0.34 ± 0.09 m) after FIRS bathing than after sitting with NO SAUNA (0.32 ± 0.0 m). After sauna heart rate was higher (p < 0.05) in TRAD (92 ± 13 beats/min) than in FIRS (71 ± 7 beats/min). In conclusion, deep penetration of infrared heat (approximately 3-4 cm into fat tissue and neuromuscular system) with mild temperature (35-50°C), and light humidity (25-35%) during FIRS bathing appears favorable for the neuromuscular system to recover from maximal endurance performance. FIRS bathing is a very light loading for the body and provides a comfortable and relaxing experience.


Children with dry eye type conditions may report less severe symptoms than adult patients.
To compare the symptom scores of children with those of adults with the same degree of mild ocular surface signs. This study included patients with at least one ocular surface symptom and sign. Data obtained from 45 subjects aged 8.79 ± 2.95 yr (Mean ± SD) (pediatric group) and 45 adults aged 52.87 ± 10.87 yr (adult group) who were matched to each pediatric patient based on ocular surface signs were analyzed. Demographic data including age and sex; parameters of ocular surface signs including fluorescein staining score, tear film breakup time and Schirmer test score; symptom scores including ocular surface disease index (OSDI) and visual analog scale (VAS) were assessed and compared between the two groups. Significant differences emerged in sex ratio as well as in age between the two groups (P < 0.001, for both). Regarding ocular surface signs and tear film tests, the study revealed no significant differences in fluorescein staining score (1.44 ± 1.95 vs. 1.40 ± 1.68 P = 0.913), tear film break up time (4.96 ± 1.94 vs. 3.82 ± 1.85 s, P = 0.612) and Schirmer score (12.79 ± 9.05 vs. 10.11 ± 8.45 mm / 5 min, P = 0.370). Regarding dry eye symptoms, both OSDI and VAS were significantly lower in the pediatric group (P = 0.001 and < 0.001, respectively) than in the adult group. Pediatric patients with mild ocular surface damage may report fewer dry eye symptoms compared to adult patients with similar stages of ocular surface damage.


Flagellin is not a major defense elicitor in Ralstonia solanacearum cells or extracts applied to Arabidopsis thaliana.
The phytopathogenic bacterium Ralstonia solanacearum requires motility for full virulence, and its flagellin is a candidate pathogen-associated molecular pattern that may elicit plant defenses. Boiled extracts from R. solanacearum contained a strong elicitor of defense-associated responses. However, R. solanacearum flagellin is not this elicitor, because extracts from wild-type bacteria and fliC or flhDC mutants defective in flagellin production all elicited similar plant responses. Equally important, live R. solanacearum caused similar disease on Arabidopsis ecotype Col-0, regardless of the presence of flagellin in the bacterium or the FLS2-mediated flagellin recognition system in the plant. Unlike the previously studied flg22 flagellin peptide, a peptide based on the corresponding conserved N-terminal segment of R. solanacearum, flagellin did not elicit any response from Arabidopsis seedlings. Thus recognition of flagellin plays no readily apparent role in this pathosystem. Flagellin also was not the primary elicitor of responses in tobacco. The primary eliciting activity in boiled R. solanacearum extracts applied to Arabidopsis was attributable to one or more proteins other than flagellin, including species purifying at approximately 5 to 10 kDa and also at larger molecular masses, possibly due to aggregation. Production of this eliciting activity did not require hrpB (positive regulator of type III secretion), pehR (positive regulator of polygalacturonase production and motility), gspM (general secretion pathway), or phcA (LysR-type global virulence regulator). Wild-type R. solanacearum was virulent on Arabidopsis despite the presence of this elicitor in pathogen extracts.


LHRH Agonists for the Treatment of Prostate Cancer: 2012.
The most recent guidelines on prostate cancer screening from the American Urological Association (2009), the National Comprehensive Cancer Network (2011), and the European Association of Urology (2011), as well as treatment and advances in disease monitoring, have increased the androgen deprivation therapy (ADT) population and the duration of ADT usage as the first-line treatment for metastatic prostate cancer. According to the European Association of Urology, gonadotropin-releasing hormone (GnRH) agonists have become the leading therapeutic option for ADT because they avoid the physical and psychological discomforts associated with orchiectomy. However, GnRH agonists display several shortcomings, including testosterone (T) surge ("clinical flare") and microsurges. T surge delays the intended serologic endpoint of T suppression and may exacerbate clinical symptoms. Furthermore, ADT manifests an adverse-event spectrum that can impact quality of life with its attendant well-documented morbidities. Strategies to improve ADT tolerability include a holistic management approach, improved diet and exercise, and more specific monitoring to detect and prevent T depletion toxicities. Intermittent ADT, which allows hormonal recovery between treatment periods, has become increasingly utilized as a methodology for improving quality of life while not diminishing chronic ADT efficacy, and may also provide healthcare cost savings. This review assesses the present and potential future role of GnRH agonists in prostate cancer and explores strategies to minimize the adverse-event profile for patients receiving ADT.


Circumferential resection margin involvement in stage III rectal cancer patients treated with curative resection followed by chemoradiotherapy: a surrogate marker for local recurrence?
Circumferential resection margin (CRM) involvement is a well-known predictor for poor prognosis in rectal cancer. However, the significance is controversial in some studies. Accordingly, this study attempted to examine the prognostic impact of CRM involvement in stage III rectal cancer. Between January 1990 and December 2007, a total of 449 patients who underwent curative resection followed by complete adjuvant chemoradiotherapy for stage III rectal cancer located within 12 cm from the anal verge were selected. Patients were divided into a CRM-positive group (n=79, 17.6%) and a CRM-negative group (n=370, 82.4%). With a median follow-up of 56.6 months, recurrent disease was seen in 53.2 and 43.5% of the CRM-positive and CRM-negative group, respectively. CRM involvement was an independent prognostic factor for 5-year systemic recurrence-free survival (HR: 1.5, CI: 1.0-2.2, p=0.017). However, no significant difference was observed for local recurrence rate between the two groups (13.0 and 13.5%, respectively, p=0.677). In this study, local recurrence rate did not differ according to CRM involvement status in stage III rectal cancer patients, although CRM involvement was shown to be an independent poor prognostic factor. Accordingly, validation of the results of this study by further large prospective randomized trials is warranted.


Nuclear localization of human AP endonuclease 1 (HAP1/Ref-1) associates with prognosis in early operable non-small cell lung cancer (NSCLC).
The present study examined the immunohistochemical expression of human AP endonuclease 1 (HAP1/Ref-1), the major endonuclease in the repair of apurinic/apyrimidinic (AP) sites in cellular DNA, in normal lung and lung carcinomas. Cellular expression of HAP1 was determined using a standard avidin-biotin-peroxidase complex (ABC) technique and an anti-HAP1 rabbit polyclonal antibody on paraffin-embedded tissue sections from normal lung and in 103 primary non-small cell lung carcinomas (NSCLCs). In normal lung, the staining for HAP1 was found to be both nuclear and cytoplasmic in the pneumocytes of the alveoli. Superficial ciliated cells of the bronchial epithelium presented cytoplasmic staining, while staining for the basal cells was mostly nuclear. Bronchial glandular cells demonstrated mixed nuclear and cytoplasmic staining. Lung carcinomas showed all patterns of expression for HAP1. Loss of HAP1 expression was associated with low proliferation index (p=0.01) and with squamous histology (p=0.04). In squamous carcinomas, a significant correlation was observed between positive nuclear HAP1 and negative p53 expression (p=0.03). A survival benefit was seen in patients presenting nuclear HAP1 expression and those presenting the nuclear HAP1+/p53- phenotype (p=0.01 and 0.007, respectively). It is concluded that nuclear HAP1 localization may be relevant to its role as a DNA repair protein and/or to the recently proposed role as an activator of wild-type p53, and thus to the better outcome seen in this group of patients.


Community analysis of pigment patterns from 37 microalgae strains reveals new carotenoids and porphyrins characteristic of distinct strains and taxonomic groups.
Phytoplankton, with an estimated 30 000 to 1 000 000 species clustered in 12 phyla, presents a high taxonomic and ecophysiological diversity, reflected by the complex distribution of pigments among the different algal classes. High performance liquid chromatography is the gold standard method for qualitative and quantitative analysis of phytoplankton pigments in seawater and culture samples, but only a few pigments can be used as robust chemotaxonomic markers. A major challenge is thus to identify new ones, characteristic of a strain, species, class or taxon that cannot be currently identified on the basis of its pigment signature. Using an optimized extraction process coupled to a HPLC de-replication strategy, we examined the pigment composition of 37 microalgae strains, representative of the broad taxonomic diversity of marine and freshwater species (excluding cyanobacteria). For each species, the major pigments already described were unambiguously identified. We also observed the presence of several minor unidentified pigments in each chromatogram. The global analysis of pigment compositions revealed a total of 124 pigments, including 98 pigments or derivatives unidentified using the standards. Absorption spectra indicated that 35 corresponded to chlorophyll/porphyrin derivatives, 57 to carotenoids and six to derivatives having both spectral signatures. Sixty-one of these unidentified or new carotenoids and porphyrin derivatives were characteristic of particular strains or species, indicating their possible use as highly specific chemotaxonomic markers capable of identifying one strain out of the 37 selected. We developed a graphical analysis using Gephi software to give a clear representation of pigment communities among the various phytoplankton strains, and to reveal strain-characteristic and shared pigments. This made it possible to reconstruct the taxonomic evolution of microalgae classes, on the basis of the conservation, loss, and/or appearance of pigments.


Neural responses to ambiguity involve domain-general and domain-specific emotion processing systems.
Extant research has examined the process of decision making under uncertainty, specifically in situations of ambiguity. However, much of this work has been conducted in the context of semantic and low-level visual processing. An open question is whether ambiguity in social signals (e.g., emotional facial expressions) is processed similarly or whether a unique set of processors come on-line to resolve ambiguity in a social context. Our work has examined ambiguity using surprised facial expressions, as they have predicted both positive and negative outcomes in the past. Specifically, whereas some people tended to interpret surprise as negatively valenced, others tended toward a more positive interpretation. Here, we examined neural responses to social ambiguity using faces (surprise) and nonface emotional scenes (International Affective Picture System). Moreover, we examined whether these effects are specific to ambiguity resolution (i.e., judgments about the ambiguity) or whether similar effects would be demonstrated for incidental judgments (e.g., nonvalence judgments about ambiguously valenced stimuli). We found that a distinct task control (i.e., cingulo-opercular) network was more active when resolving ambiguity. We also found that activity in the ventral amygdala was greater to faces and scenes that were rated explicitly along the dimension of valence, consistent with findings that the ventral amygdala tracks valence. Taken together, there is a complex neural architecture that supports decision making in the presence of ambiguity: (a) a core set of cortical structures engaged for explicit ambiguity processing across stimulus boundaries and (b) other dedicated circuits for biologically relevant learning situations involving faces.


First Report of Late Blight Caused by Phytophthora infestans Clonal Lineage US-24 on Potato (Solanum tuberosum) in Wisconsin.
Potato (Solanum tuberosum) crops are grown on over 25,090 ha in Wisconsin annually. Late blight, caused by Phytophthora infestans (Mont.) deBary, is a potentially devastating disease that affects tomato and potato crops in Wisconsin every few years when inoculum is introduced and weather conditions favor disease. Incidence and severity of late blight are highly variable in these few years due to differences in pathogen clonal lineages, their timing and means of introduction, and weather conditions. Prevention of this disease through prophylactic fungicide application can cost producers millions of dollars annually in additional chemical, fuel, and labor expenses. Populations of P. infestans in the U.S. have recently undergone significant genetic change, resulting in isolates with unique clonal lineages and epidemiological characteristics (1). In 2010, late blight epidemics were of low severity in discrete portions of a few fields and were seen exclusively on potato in two counties of central Wisconsin. Symptoms included water-soaked to dark brown circular lesions with pale green haloes accompanied by white fuzzy pathogen sporulation typically on leaf undersides in high humidity conditions. Infected plants were collected by professional crop consultants and submitted to the authors at the University of Wisconsin Vegetable Pathology Laboratory in Madison, Wisconsin. Eight isolates of P. infestans were generated from individual leaf samples, representing separate fields, by removing sporangia from sporulating lesions and placing onto Rye A agar amended with rifampicin and ampicillin. Axenic, single zoospore-derived cultures of isolates were generated from parent cultures and maintained on Rye A agar for further characterization. Mycelium was coenocytic with hyphal diameter of 5 to 8 μm (n = 50). Sporangia were limoniform to ovoid, semi- to fully papillate, caducous, had short pedicels, and were 36.22 × 19.11 μm (height × width; n = 50). The average length-width ratio was 1.91. Allozyme banding patterns at the glucose-6-phosphate isomerase (Gpi) locus indicated a 100/100/111 profile, consistent with the US-24 clonal lineage (3,4). Mating type assays confirmed the isolates to be A1 and intermediate insensitivity to mefenoxam was observed in vitro (4). Genomic DNA was extracted with a phenol:chloroform:isoamyl alcohol solution and restriction fragment length polymorphism (RFLP) analysis was performed using the RG-57 probe on a representative isolate and resulted in banding patterns consistent with US-24 (2,3). Clonal lineages of P. infestans documented in Wisconsin in previous epidemics included US-8 in the mid-1990s and US-1 in the 1970s. The US-24 (A1) clonal lineage was very widespread in the U.S. in 2010 and its presence in Wisconsin in the same year as identification of US-22 (A2) posed great concern for potential sexual recombination, oospore production, and soil persistence. Fortunately, the opposite mating types were separated spatiotemporally. To the best of our knowledge, this is the first report of the P. infestans clonal lineage US-24 causing late blight on potato in Wisconsin. References: (1) K. Deahl. (Abstr.) Phytopathology 100:S161, 2010. (2) S. B. Goodwin et al. Curr. Genet. 22:107, 1992. (3) Hu et al. Plant Dis. 96:1323, 2012. (4) A. C. Seidl and A. J. Gevens. (Abstr.) Phytopathology 101:S162, 2011.


Parvicapsula bicornis n. sp. and P. limandae n. sp. (Myxozoa, Parvicapsulidae) in Pleuronectidae (Teleostei, Heterosomata) from Denmark.
Two species of Parvicapsula were found in the kidney tubules and the urinary bladder of 2 pleuronectid fish from the northern Oresund, Denmark. The coelozoic, spherical, disporic trophozoites of both species are 10 to 12 pm in diameter. The myxospores of both species are elongate, asymmetrical and slightly curved, and have spherical polar capsules. Parvicapsula bicornis n. sp. (6-8 x 5-6 microm, polar capsule 2.5 microm in diameter) occurs in Pleuronectes platessa. The polar capsules of P. bicornis are arranged symmetrically on either side of the longitudinal axis and its spores differ from other species of Parvicapsula in having two 2-3 microm long posterior processes of different length. Parvicapsula limandae n. sp. (8-11 x 4-5 pm, polar capsule 1.6 microm in diameter) is found in Limanda limanda. The polar capsules are arranged along the longitudinal axis. It differs from Parvicapsula unicornis Kabata, 1962, recorded from L. limanda, in the arrangement of the polar capsules and in the absence of a posterior horn-like projection. The phylogenetic relationship between P. bicornis n. sp., P. limandae n. sp. and other Parvicapsula spp. was examined with their partial small subunit rDNA (SSU rDNA) sequences. P. limandae n. sp. and P. asymmetrica appear to be closely related, while P. bicornis n. sp. and P. minibicornis are the most divergent members of the genus.


Pharmacological approaches to improve endothelial repair mechanisms.
Endothelial injury is thought to play a pivotal role in the development and progression of vascular diseases, such as atherosclerosis, hypertension or restenosis, as well as their complications, including myocardial infarction or stroke. Accumulating evidence suggests that bone marrow-derived endothelial progenitor cells (EPCs) promote endothelial repair and contribute to ischemia-induced neovascularization. Coronary artery disease and its risk factors, such as diabetes, hypercholesterolemia, hypertension and smoking, are associated with a reduced number and impaired functional activity of circulating EPCs. Moreover, initial data suggest that reduced EPC levels are associated with endothelial dysfunction and an increased risk of cardiovascular events, compatible with the concept that impaired EPC-mediated vascular repair promotes progression of vascular disease. In this review we summarize recent data on the effects of pharmacological agents on mobilization and functional activity of EPCs. In particular, several experimental and clinical studies have suggested that statins, angiotensin-converting enzyme inhibitors, angiotensin II type 1 receptor blockers, PPAR-gamma agonists and erythropoietin increase the number and functional activity of EPCs. The underlying mechanisms remain largely to be defined; however, they likely include activation of the PI3-kinase/Akt pathway and endothelial nitric oxide synthase, as well as inhibition of NAD(P)H oxidase activity of progenitor cells.


Physical activity and incidence of hypertension in college alumni.
In a study population of 14,998 Harvard male alumni, 681 hypertensives were first diagnosed during a 6-10-year follow-up beginning 16-50 years after college entrance. The study comprised 105,662 man-years of observation of these men who had entered college in 1916-1950, and who were followed from 1962 or 1966 to 1972. Presence or absence of a background of collegiate sports did not influence risk of hypertension in this study population, nor did stair-climbing, walking, or light sports play by alumni. But, alumni who did not engage in vigorous sports play were at 35% greater risk of hypertension than those who did, and this relationship held at all ages, 35-74 years. Higher levels of body mass index, weight gain since college, history of parental hypertension, and lack of strenuous exercise independently predicted increased risk of hypertension in alumni. Men 20% or more over ideal weight-for-height were at 78% greater risk than lighter men. Those who had gained 25+ lbs (c. 11.5+ kg) since entering college were at 60% greater risk than those who had gained less. Alumni with a hypertensive parent were at 83% higher risk than men without such parentage. Contemporary vigorous exercise was inversely related to hypertension risk, but chiefly among alumni overweight-for-height. In the clinical sense, attributable risk estimates ranged from 30% to nearly 50% for the alumni characteristics of overweight, weight gain, parental hypertension, and lack of vigorous exercise. In the community sense, attributable risk of these same characteristics ranged 13-26%. To sum up, vigorous exercise is associated with lower hypertension incidence, and, without necessarily altering body weight-for-height, avoids or reduces fat and promotes muscle; obesity, rather than excess weight-for-height, is associated with higher hypertension incidence; hence, vigorous exercise is appropriate for use as an intervention regimen in the prevention of hypertension.


Osteoporosis management: a perspective based on bisphosphonate data from randomised clinical trials and observational databases.
The efficacy of treatments for osteoporosis can be evaluated using a variety of study designs. This article aims to comprehensively review the evidence for bisphosphonate anti-fracture efficacy in postmenopausal women, discussing the strengths and limitations associated with each study method. Literature analysis included English-language publications reporting results of randomised controlled trials (RCTs), post hoc analyses, meta analyses and observational studies evaluating the efficacy of alendronate (ALN), ibandronate (IBN), risedronate (RIS) and zoledronate (ZOL), with an initial sample size > or = 100 patients, and follow-up data for at least 1 year. Primary and secondary analyses of RCT data suggest differences among bisphosphonates with regard to site-specific anti-fracture efficacy and onset of fracture risk reduction. While some observational studies indicate differences in clinical outcomes among these agents, others report similar effectiveness. ALN and RIS data demonstrate sustained fracture protection for up to 10 and 7 years of treatment respectively. The efficacy of IBN and ZOL has been evaluated for up to 3 and 5 years respectively. Understanding of the benefits of bisphosphonate treatment can be maximised by evaluating complementary data from RCTs and observational database studies. Fracture risk reduction with bisphosphonates is shown in RCTs and in real-world clinical settings.


Breath tests sustainability in hospital settings: cost analysis and reimbursement in the Italian National Health System.
The high demand of Breath Tests (BT) in many gastroenterological conditions in time of limited resources for health care systems, generates increased interest in cost analysis from the point of view of the delivery of services to better understand how use the money to generate value. This study aims to measure the cost of C13 Urea and other most utilized breath tests in order to describe key aspects of costs and reimbursements looking at the economic sustainability for the hospital. A hospital based cost-analysis of the main breath tests commonly delivery in an ambulatory setting is performed. Mean salary for professional nurses and gastroenterologists, drugs/preparation used and disposable materials, purchase and depreciation of the instrument and the testing time was used to estimate the cost, while reimbursements are based on the 2013 Italian National Health System ambulatory pricelist. Variables that could influence the model are considered in the sensitivity analyses. The mean cost for C13--Urea, Lactulose and Lactose BT are, respectively, Euros 30,59; 45,20 and 30,29. National reimbursement often doesn't cover the cost of the analysis, especially considering the scenario with lower number of exam. On the contrary, in high performance scenario all the reimbursement could cover the cost, except for the C13 Urea BT that is high influenced by the drugs cost. However, consideration about the difference between Italian Regional Health System ambulatory pricelist are done. Our analysis shows that while national reimbursement rates cover the costs of H2 breath testing, they do not cover sufficiently C13 BT, particularly urea breath test. The real economic strength of these non invasive tests should be considered in the overall organization of inpatient and outpatient clinic, accounting for complete diagnostic pathway for each gastrointestinal disease.


An expanded self-antigen peptidome is carried by the human lymph as compared to the plasma.
The pre-nodal afferent lymph is the fluid which directly derives from the extracellular milieu from every parenchymal organ and, as it continues to circulate between the cells, it collects products deriving from the organ metabolism/catabolism. A comprehensive qualitative and quantitative investigation of the self-antigenic repertoire transported by the human lymph is still missing. A major difference between lymph and plasma could be visualized by FPLC and 2D gel in the amount of low molecular weight products corresponding to peptide fragments. Naturally processed peptides in normal pre-nodal human lymph were then fractionated by HPLC and characterized by multidimensional mass spectrometry. Analysis of more then 300 sequences identified self-peptides derived from both intracellular and extracellular proteins revealing the variety of catabolic products transported by human lymph. Quantitative analysis established that at least some of these peptides are present in the circulating lymph in nanomolar concentration. The peptidome, generated by physiological tissue catabolism and transported by the pre-nodal lymph, is in addition to the self-peptidome generated in endosomal compartment. Unlike self antigen processed by local or nodal APC, which mostly produce epitopes constrained by the endosomal processing activity, self antigens present in the lymph could derived from a wider variety of processing pathways; including caspases, involved in cellular apoptosis, and ADAM and other metalloproteinases involved in surface receptor editing, cytokines processing and matrix remodeling. Altogether, expanding the tissue-specific self-repertoire available for the maintenance of immunological tolerance.


The PAK system links Rho GTPase signaling to thrombin-mediated platelet activation.
Regulation of the platelet actin cytoskeleton by the Rho family of small GTPases is essential for the proper maintenance of hemostasis. However, little is known about how intracellular platelet activation from Rho GTPase family members, including Rac, Cdc42, and Rho, translate into changes in platelet actin structures. To better understand how Rho family GTPases coordinate platelet activation, we identified platelet proteins associated with Rac1, a Rho GTPase family member, and actin regulatory protein essential for platelet hemostatic function. Mass spectrometry analysis revealed that upon platelet activation with thrombin, Rac1 associates with a set of effectors of the p21-activated kinases (PAKs), including GIT1, βPIX, and guanine nucleotide exchange factor GEFH1. Platelet activation by thrombin triggered the PAK-dependent phosphorylation of GIT1, GEFH1, and other PAK effectors, including LIMK1 and Merlin. PAK was also required for the thrombin-mediated activation of the MEK/ERK pathway, Akt, calcium signaling, and phosphatidylserine (PS) exposure. Inhibition of PAK signaling prevented thrombin-induced platelet aggregation and blocked platelet focal adhesion and lamellipodia formation in response to thrombin. Together, these results demonstrate that the PAK signaling system is a key orchestrator of platelet actin dynamics, linking Rho GTPase activation downstream of thrombin stimulation to PAK effector function, MAP kinase activation, calcium signaling, and PS exposure in platelets.


RNA deep sequencing reveals novel candidate genes and polymorphisms in boar testis and liver tissues with divergent androstenone levels.
Boar taint is an unpleasant smell and taste of pork meat derived from some entire male pigs. The main causes of boar taint are the two compounds androstenone (5α-androst-16-en-3-one) and skatole (3-methylindole). It is crucial to understand the genetic mechanism of boar taint to select pigs for lower androstenone levels and thus reduce boar taint. The aim of the present study was to investigate transcriptome differences in boar testis and liver tissues with divergent androstenone levels using RNA deep sequencing (RNA-Seq). The total number of reads produced for each testis and liver sample ranged from 13,221,550 to 33,206,723 and 12,755,487 to 46,050,468, respectively. In testis samples 46 genes were differentially regulated whereas 25 genes showed differential expression in the liver. The fold change values ranged from -4.68 to 2.90 in testis samples and -2.86 to 3.89 in liver samples. Differentially regulated genes in high androstenone testis and liver samples were enriched in metabolic processes such as lipid metabolism, small molecule biochemistry and molecular transport. This study provides evidence for transcriptome profile and gene polymorphisms of boars with divergent androstenone level using RNA-Seq technology. Digital gene expression analysis identified candidate genes in flavin monooxygenease family, cytochrome P450 family and hydroxysteroid dehydrogenase family. Moreover, polymorphism and association analysis revealed mutation in IRG6, MX1, IFIT2, CYP7A1, FMO5 and KRT18 genes could be potential candidate markers for androstenone levels in boars. Further studies are required for proving the role of candidate genes to be used in genomic selection against boar taint in pig breeding programs.


Newly designed hemagglutinin-Der p 2 chimera is a potential candidate for allergen specific immunotherapy.
Aim To investigate the immunomodulatory potential of a chimera composed of the receptor-binding domain of hemagglutinin 1 (H1s) from Influenza virus and Der p 2 (D2) allergen for allergen-specific immunotherapy of house-dust mite allergy (HDM). H1sD2 chimera and D2 allergen were produced by genetic engineering in E. coli. Recombinant antigens were extracted from inclusion bodies by urea, then refolded and purified by immobilized- metal affinity chromatography (IMAC). Purity was verified by 2D-PAGE and secondary structures were assessed by CD spectroscopy. IgE reactivity of H1sD2 and D2 was tested in western blot with sera from 8 persons with clinical history of HDM allergy. Immunogenicity of H1sD2 and D2 were analyzed in Balb/c mice. Cytokine profile was analyzed by ELISA after stimulation of mouse spleen cells with H1sD2 and D2. Leukocyte population abundance of cells isolated from spleen and lymph node was assessed by flow cytometry. Purified recombinant proteins H1sD2 (42 kDa) and D2 (15 kDa) revealed well defined secondary structures, and preserved IgE reactive epitopes. Analysis of supernatants of mouse spleen cells after stimulation with H1sD2 and D2, revealed a qualitatively different cytokine profile from H1sD2 immunized mouse cells (increase in IL10). CD8+ cells were decreased in the lymph node of D2 immunized mice, whereas H1sD2 immunization led to an increase of CD8+ cells in both the lymph node and the spleen. H1sD2 chimera attenuates Der p 2-inherent Th2 response and directs the immune response toward Th1 and Treg phenotype.


Successful engraftment of human postnatal thymus in severe combined immune deficient (SCID) mice: differential engraftment of thymic components with irradiation versus anti-asialo GM-1 immunosuppressive regimens.
To develop a model of human thymus growth in vivo, we have implanted postnatal human thymus under the renal capsule of severe combined immune deficient (SCID) mice and assayed for graft survival and graft characteristics 1-3 mo after engraftment. Three groups of SCID mice were engrafted with postnatal human thymus: untreated SCID mice, SCID mice pretreated with 400 cGy of gamma irradiation 1-5 d before engraftment, and SCID mice treated with intraperitoneal anti-asialo GM-1 antiserum every 4-5 d during engraftment. In the untreated group of SCID mice, only 37% of grafts survived and consisted of human thymic microenvironment components and human immature thymocytes. Irradiation of SCID mice before engraftment improved survival of human thymic grafts to 83%, but these grafts were largely devoid of thymocytes and contained only thymic microenvironment components with large numbers of thymic macrophages. Treatment of SCID mice with anti-asialo GM-1 antiserum throughout the engraftment period also promoted human thymus engraftment (70%) and induced SCID B cell Ig production (SCID[Ig+]) in 38% of animals. In SCID(Ig-) anti-asialo GM-1-treated mice, the human thymic grafts were similar in content to those in untreated SCID mice. However, in anti-asialo GM-1-treated animals with grafts that became SCID(Ig+), all animals were found to have mouse-human chimeric grafts in that the human thymic microenvironment (human fibroblasts, thymic epithelium, vessels) was colonized by murine T cells. These data demonstrate that human postnatal thymus will grow as xenografts in SCID mice, and that the components of human thymus that engraft are dependent on the immunosuppressive regimen used in recipient mice. A striking finding in this study was the induction of T and B lymphopoiesis in SCID mice by abrogation of NK cell activity with in vivo anti-asialo GM-1 treatment. These data strongly suggest that asialo GM-1+ NK cells and/or macrophages play a role in mediation of suppression of lymphopoiesis in SCID mice.


A brand new START: abscisic acid perception and transduction in the guard cell.
The soluble receptors of abscisic acid (ABA) have been identified in Arabidopsis thaliana. The 14 proteins in this family, bearing the double name of PYRABACTIN RESISTANCE/PYRABACTIN-LIKE (PYR/PYL) or REGULATORY COMPONENTS OF ABA RECEPTOR (RCAR) (collectively referred to as PYR/PYL/RCAR), contain between 150 and 200 amino acids with homology to the steroidogenic acute regulatory-related lipid transfer (START) protein. Structural studies of these receptors have provided rich insights into the early mechanisms of ABA signaling. The binding of ABA to PYR/PYL/RCAR triggers the pathway by inducing structural changes in the receptors that allows them to sequester members of the clade A negative regulating protein phosphatase 2Cs (PP2Cs). This liberates the class III ABA-activated Snf1-related kinases (SnRK2s) to phosphorylate various targets. In guard cells, a specific SnRK2, OPEN STOMATA 1 (OST), stimulates H(2)O(2) production by NADPH oxidase respiratory burst oxidase protein F and inhibits potassium ion influx by the inward-rectifying channel KAT1. OST1, the kinase CPK23, the calcium-dependent kinase CPK21, and the counteracting PP2Cs modulate the slow anion channel SLAC1, a pathway that contributes to stomatal responses to diverse stimuli, including ABA and carbon dioxide. A minimal ABA response pathway that leads to activation of the SLAC1 homolog, SLAH3, and presumably stomatal closure has been reconstituted in vitro. The identification of the soluble receptors and core components of the ABA signaling pathway provides promising targets for crop design with higher resilience to water deficit while maintaining biomass.


American Society of Clinical Oncology policy statement: opportunities in the patient protection and affordable care act to reduce cancer care disparities.
Patients in specific vulnerable population groups suffer disproportionately from cancer. The elimination of cancer disparities is critically important for lessening the burden of cancer. The Patient Protection and Affordable Care Act provides both opportunities and challenges for addressing cancer care disparities and access to care. The American Society of Clinical Oncology (ASCO) advocates for policies that ensure access to cancer care for the underserved. Such policies include insurance reform and the reduction of economic barriers to quality health care. Building on ASCO's prior statement on disparities in cancer care (2009), this article summarizes elements of the health care law that are relevant to cancer disparities and provides recommendations for addressing major provisions in the law. It outlines specific strategies to address insurance reform, access to care, quality of care, prevention and wellness, research on health care disparities, and diversity in the health care workforce. ASCO is committed to leading efforts toward the improvement of cancer care among the most vulnerable patients.