Lipid vesicle size of an oral influenza vaccine delivery vehicle influences the Th1/Th2 bias in the immune response and protection against infection.
Previous studies, using parenteral administration of antigen in lipid vesicles, have indicated a possible role for vesicle size in determining the Th1/Th2 bias of the resulting immune response. We have also demonstrated that the incorporation of bile salts into lipid vesicles (bilosomes) allows successful induction of mucosal and systemic immunity via the oral route. The following study was therefore carried out to determine whether size could also influence the Th1/Th2 bias in the immune response to bilosome entrapped influenza A antigen containing haemagglutinin administered by the oral route in the mouse and whether this could influence the disease process in the classical ferret model of disease. Consequently we produced two formulations of influenza A antigen entrapped in bilosomes: BV3 which contained a single population (range 10-100 nm, Z-average diameter 250 nm) and BV which had two populations (60-350 and 400-2,500 nm, Z-average 980 nm). Following oral vaccination of BALB/c mice, BV was found to generate an immune response that had a significantly greater Th1 bias than BV3 as measured by serum IgG2a production and antigen-induced spleen cell IFN-gamma production. In the traditional infection challenge model (ferrets) vaccination with BV (large) vesicles resulted in greater protection in terms of symptom-score and a higher responder number. However, both oral vaccine formulations were an improvement on intramuscular administration in terms of higher antibody production, lower temperatures, and reduced symptoms over time, post-infection. The results presented here demonstrate that oral vaccine formulations can be physically modified to manipulate resultant immune responses following vaccination and consequently can be designed to enhance the effectiveness of candidate vaccine antigens.


Potential mechanisms of attenuation for rifampicin-passaged strains of Flavobacterium psychrophilum.
Flavobacterium psychrophilum is the etiologic agent of bacterial coldwater disease in salmonids. Earlier research showed that a rifampicin-passaged strain of F. psychrophilum (CSF 259-93B.17) caused no disease in rainbow trout (Oncorhynchus mykiss, Walbaum) while inducing a protective immune response against challenge with the virulent CSF 259-93 strain. We hypothesized that rifampicin passage leads to an accumulation of genomic mutations that, by chance, reduce virulence. To assess the pattern of phenotypic and genotypic changes associated with passage, we examined proteomic, LPS and single-nucleotide polymorphism (SNP) differences for two F. psychrophilum strains (CSF 259-93 and THC 02-90) that were passaged with and without rifampicin selection. Rifampicin resistance was conveyed by expected mutations in rpoB, although affecting different DNA bases depending on the strain. One rifampicin-passaged CSF 259-93 strain (CR) was attenuated (4 % mortality) in challenged fish, but only accumulated eight nonsynonymous SNPs compared to the parent strain. A CSF 259-93 strain passaged without rifampicin (CN) accumulated five nonsynonymous SNPs and was partially attenuated (28 % mortality) compared to the parent strain (54.5 % mortality). In contrast, there were no significant change in fish mortalities among THC 02-90 wild-type and passaged strains, despite numerous SNPs accumulated during passage with (n = 174) and without rifampicin (n = 126). While only three missense SNPs were associated with attenuation, a Ser492Phe rpoB mutation in the CR strain may contribute to further attenuation. All strains except CR retained a gliding motility phenotype. Few proteomic differences were observed by 2D SDS-PAGE and there were no apparent changes in LPS between strains. Comparative methylome analysis of two strains (CR and TR) identified no shared methylation motifs for these two strains. Multiple genomic changes arose during passage experiments with rifampicin selection pressure. Consistent with our hypothesis, unique strain-specific mutations were detected for the fully attenuated (CR), partially attenuated (CN) and another fully attenuated strain (B17).


p16 Immunohistochemistry Interpretation by Nonpathologists as an Accurate Method for Diagnosing Cervical Precancer and Cancer.
We conducted a pilot study of whether nonpathologists could accurately diagnose cervical precancer in biopsies using only a basic light microscope, evaluating p16 immunohistochemistry (p16 IHC) of biopsies, and video-based training for both. Using biopsies collected as part of a screening study conducted in rural China, we randomly selected 50 biopsies with a precancerous diagnosis of cervical intraepithelial neoplasia grade 2 (CIN2) or more severe (CIN2+) and 50 biopsies with diagnosis of CIN less severe than CIN2, and stained them for p16 using a commercial IHC kit. Twelve nonpathologists of varying educational backgrounds living in Beijing, China received video training and were assigned one of 4 sets of 25 CIN2+ and 25 CIN less severe than CIN2 for evaluation. A pathologist reviewed all 100 cases. The mean sensitivity and specificity of the p16 IHC staining scored by the nonpathologists were 91.7% and 94.1%, respectively, compared to scoring by the pathologist. The readers and the pathologist agreed on p16 IHC scoring for 42 (84%) of the 50 slides of CIN less severe than CIN2 and 37 (74%) of the 50 CIN2+ slides. The mean sensitivity and specificity for consensus CIN2+ of p16 IHC as scored by the readers were 88% and 87%, respectively, versus an overall sensitivity and specificity by the pathologist of 96% and 92%, respectively. We demonstrated that nonpathologists can accurately diagnose CIN2+ using p16 IHC alone.


A longitudinal modelling study estimates acute symptoms of community acquired pneumonia recover to baseline by 10 days.
Our aims were to address three fundamental questions relating to the symptoms of community-acquired pneumonia (CAP): Do patients completely recover from pneumonia symptoms? How long does this recovery take? Which factors influence symptomatic recovery?We prospectively recruited patients at two hospitals in Liverpool, UK, into a longitudinal, observational cohort study and modelled symptom recovery from CAP. We excluded patients with cancer, immunosuppression or advanced dementia, and those who were intubated or palliated from admission. We derived a statistical model to describe symptom patterns.We recruited 169 (52% male) adults. Multivariable analysis demonstrated that the time taken to recover to baseline was determined by the initial severity of symptoms. Severity of symptoms was associated with comorbidity and was inversely related to age. The pattern of symptom recovery was exponential and most patients' symptoms returned to baseline by 10 days.These results will inform the advice given to patients regarding the resolution of their symptoms. The recovery model described here will facilitate the use of symptom recovery as an outcome measure in future clinical trials.


Pig coronary artery smooth muscle: substrate and pH dependence of the two calcium pumps.
The properties of the pig coronary artery smooth muscle Ca transport mechanisms and the effects of pH were examined. The Ca pump in the plasma membrane requires less than 50 microM Mg-ATP2- for half-maximal activity, but the pump in the endoplasmic reticulum requires greater than 700 microM Mg-ATP2-. The Ca uptake by the various subcellular organelles occurs with the following affinity characteristics towards Ca2+: plasma membrane, Km = 0.91 +/- 0.06 microM, and Hill coefficient = 1.31 +/- 0.09; endoplasmic reticulum, Km = 0.58 +/- 0.05 microM, and Hill coefficient = 2.48 +/- 0.34; and mitochondria, Km = 7.1 +/- 1.16, and Hill coefficient = 0.93 +/- 0.10. The active Ca transport systems showed similar pH dependence in that the Ca uptake at pH 6.8 was greater than at 7.6. However, the pH effects were much larger on the Ca uptake by the endoplasmic reticulum and mitochondrial membranes than by the plasma membrane. The plasma membrane also showed a small amount of pH-dependent, high-affinity Ca binding. Ca efflux from passively loaded plasma membrane vesicles was similar at pH 6.4 and 7.4. From these data, it is concluded that the endoplasmic reticulum Ca pump may play a larger role in the vasodilation of large coronary artery, but the role of the plasma membrane may be more in maintenance of relaxation mechanism.


Hospital environment, nurse-physician relationships and quality of care: questionnaire survey.
This paper is a report of a study conducted to (a) to compare hospitalized patients' and nurses' perceptions of the hospital environment, nurse-physician relationships and quality of care; (b) to determine which factors best predict the quality of care from hospitalized patients' and nurses' perspectives; and (c) to assess the relationships among all variables. Quality of care is a function of many factors and includes elements of the hospital environment and nurse-physician relationships. However, comparisons between patients' and nurses' perceptions are relatively limited. This was a cross-sectional study, and 575 patients and 220 nurses across 13 units completed questionnaires. Data were collected in 2009 and analysed using descriptive statistics, independent t-tests, stepwise regression and path analysis. Overall, patients' mean scores were statistically significantly higher than those of nurses on perception of hospital environment (3·05 vs. 2·65 points), nurse-physician relationships (7·88 vs. 6·53 points) and quality of care (7·91 vs. 6·63 points) (P < 0·001). Both the hospital environment and nurse-physician relationships were statistically significant predictors of quality of care. Path analysis showed that quality of care was affected by the hospital environment, nurse-physician relationships and years of education for patients (P < 0·05). However, for nurses, quality of care was only directly affected by the hospital environment and nurse-physician relationships (P < 0·05). These findings may help nursing leaders to ensure future excellent patient care by helping them to see which areas need improvement. They may also help nursing leaders to develop strategies to meet patients' and nurses' expectations.


Plutonium-induced proliferative lesions and pulmonary epithelial neoplasms in the rat: immunohistochemical and ultrastructural evidence for their origin from type II pneumocytes.
Immunohistochemistry and transmission electron microscopy were used to clarify the cellular origin for plutonium-239-induced pulmonary proliferative (preneoplastic) epithelial lesions and epithelial neoplasms in F344 rats. Examples of each histologic type of proliferative lesion and neoplasm were stained by the avidin-biotin complex immunoperoxidase method using antibodies to rat surfactant apoprotein and Clara cell antigen. Rat surfactant apoprotein immunostaining was detected in type II pneumocytes in sections of normal lung, in the cells of the proliferative lesions classified histologically as alveolar epithelial hyperplasia (51) and mixed foci (alveolar epithelial hyperplasia with fibrosis) (30), and in adenomas (2), adenocarcinomas (3), and adenosquamous carcinomas (2). With the exception of one adenosquamous carcinoma, Clara cell antigen immunostaining was not detected in any of the pulmonary lesions but was detected in nonciliated cuboidal epithelial (Clara) cells in normal bronchioles. The epithelial cells of the proliferative lesions and neoplasms had ultrastructural features consistent with type II pneumocytes, i.e., the presence of cytoplasmic lamellar and multivesicular bodies. The results of these studies indicate that the majority of plutonium-induced proliferative epithelial lesions and neoplasms in the rat originate from alveolar type II pneumocytes.


Predicting observed and hidden extreme events in complex nonlinear dynamical systems with partial observations and short training time series.
Extreme events appear in many complex nonlinear dynamical systems. Predicting extreme events has important scientific significance and large societal impacts. In this paper, a new mathematical framework of building suitable nonlinear approximate models is developed, which aims at predicting both the observed and hidden extreme events in complex nonlinear dynamical systems for short-, medium-, and long-range forecasting using only short and partially observed training time series. Different from many ad hoc data-driven regression models, these new nonlinear models take into account physically motivated processes and physics constraints. They also allow efficient and accurate algorithms for parameter estimation, data assimilation, and prediction. Cheap stochastic parameterizations, judicious linear feedback control, and suitable noise inflation strategies are incorporated into the new nonlinear modeling framework, which provide accurate predictions of both the observed and hidden extreme events as well as the strongly non-Gaussian statistics in various highly intermittent nonlinear dyad and triad models, including the Lorenz 63 model. Then, a stochastic mode reduction strategy is applied to a 21-dimensional nonlinear paradigm model for topographic mean flow interaction. The resulting five-dimensional physics-constrained nonlinear approximate model is able to accurately predict extreme events and the regime switching between zonally blocked and unblocked flow patterns. Finally, incorporating judicious linear stochastic processes into a simple nonlinear approximate model succeeds in learning certain complicated nonlinear effects of a six-dimensional low-order Charney-DeVore model with strong chaotic and regime switching behavior. The simple nonlinear approximate model then allows accurate online state estimation and the short- and medium-range forecasting of extreme events.


An assessment framework for climate-proof nature-based solutions.
Raising interest in 'nature-based solutions' (NBS) has inspired attempts to organise their principles and qualities within comprehensive and internally consistent evaluation frameworks, so as to demonstrate the superior performance of 'working with nature'. However, the proposed frameworks stop short of taking into account the changing conditions in which NBS are set to operate. Climate change, in particular, can alter ecosystems and their services, and may undermine the performance of green solutions that rely on them. We present here a 'dynamic' assessment framework that explicitly accounts for the impact of climate change on the effectiveness of the proposed NBS. The framework is based on an innovative approach that integrates system analysis and backcasting. Although it has not yet been applied to the NBS context, backcasting is well-suited to seize the transformational character of NBS, as it encourages 'breakthrough' leaps rather than incremental improvements. Our framework factors in the multifunctional character of NBS and is designed to capture associated direct benefits/costs and co-benefits/costs. It is meant to be applied ex ante to ideally support the choice between innovative NBS and traditional options, in an effort to respond to the societal challenges identified by the EU Research & Innovation agenda on the environment.


Cystine Stone Formers Have Impaired Health-Related Quality of Life Compared with Noncystine Stone Formers: A Case-Referent Study Piloting the Wisconsin Stone Quality of Life Questionnaire Among Patients with Cystine Stones.
Cystinuria is a rare cause of urolithiasis. Affected patients have an earlier onset and more aggressive disease than patients with other stone etiologies. We assessed the health-related quality of life (HRQOL) of cystine stone-forming patients using the disease-specific Wisconsin Stone Quality of Life questionnaire (WISQOL). Cystine patients treated in our stone clinics (n = 12) completed the WISQOL; information about medical and stone histories was gathered. Patients were matched with noncystine stone formers (n = 12) for gender, age, and comorbidities. In addition, a second control group (n = 90), also from our institution and consisting of mixed calcium stone formers, was included. WISQOL responses were compared between groups. Cystine patients had significantly lower total WISQOL scores than noncystine patients. Compared with noncystine stone formers, cystine stone formers also had lower HRQOL scores for subscales (domains) related to social impact, emotional impact, disease impact, and vitality (p ≤ 0.04 for all). On specific items, cystine patients reported significantly more sleep problems (p = 0.02), more bother with nocturia (p = 0.03), and feeling tired or fatigued (p = 0.02). Among those with current stones, cystine patients scored lower than noncystine patients for total score and in two of four domains. Using a stone-specific questionnaire, patients with cystine stones have lower HRQOL compared with noncystine stone formers. Identifying and addressing specific areas of decrement in patients with cystine stones may improve disease management and patients' HRQOL.


Reactive astrocytosis, microgliosis and inflammation in rats with neonatal hydrocephalus.
The deleterious effects of hydrocephalus, a disorder that primarily affects children, include reactive astrocytosis, microgliosis and inflammatory responses; however, the roles that these mechanisms play in the pathophysiology of hydrocephalus are still not clear in terms of cytopathology and gene expression. Therefore we have examined neuroinflammation at both the cellular and the molecular levels in an experimental model of neonatal obstructive hydrocephalus. On post-natal day 1, rats received an intracisternal injection of kaolin to induce hydrocephalus; control animals received saline injections. Prior to sacrifice on post-natal day 22, animals underwent magnetic resonance imaging to quantify ventricular enlargement, and the parietal cortex was harvested for analysis. Immunohistochemistry and light microscopy were performed on 5 hydrocephalic and 5 control animals; another set of 5 hydrocephalic and 5 control animals underwent molecular testing with Western blots and a gene microarray. Scoring of immunoreactivity on a 4-point ranking scale for GFAP and Iba-1 demonstrated an increase in reactive astrocytes and reactive microglia respectively in the hydrocephalic animals compared to controls (2.90±0.11 vs. 0.28±0.26; 2.91±0.11 vs. 0.58±0.23, respectively). Western blots confirmed these results. Microarray analysis identified significant (1.5-fold) changes in 1729 of 33,951 genes, including 26 genes out of 185 genes (26/185) in the cytokine-cytokine receptor interaction pathway, antigen processing and presentation pathways (15/66), and the apoptosis pathway (10/69). Collectively, these results demonstrate alterations in normal physiology and an up-regulation of the inflammatory response. These findings lead to a better understanding of neonatal hydrocephalus and begin to form a baseline for future treatments that may reverse these effects.


Mechanisms by which tumor cells and monocytes expressing the angiogenic factor thymidine phosphorylase mediate human endothelial cell migration.
The angiogenic factor thymidine phosphorylase (TP) is highly expressed in many human solid tumors, and the level of its expression is associated with tumor neovascularization, invasiveness, and metastasis and with shorter patient survival time. TP promotes endothelial cell (EC) migration in vitro and angiogenesis in vivo, and these have been linked to its enzymatic activity. The mechanism by which TP stimulates EC migration was investigated using human umbilical vein ECs (HUVECs). TP induced concentration-dependent HUVEC migration, which required a TP gradient and thymidine and which was abrogated by the TP inhibitor CIMU (5-chloro-6(1-imidazolylmethyl)uracil). The chemotactic actions of TP plus thymidine were duplicated by the TP metabolite, 2-deoxyribose-1-phosphate (dR-1-P), and 10-fold more potently by its subsequent metabolite, 2-deoxyribose (2dR). Migration induced by dR-1-P, but not 2dR, was blocked by an alkaline phosphatase inhibitor, suggesting that the actions of dR-1-P first required its conversion to 2dR. In the migration assay, [5'-3H]dThd was metabolized to dR-1-P (96%) and 2dR (3.8%), and a gradient of both metabolites was maintained between the lower and upper chambers over the entire 5-h assay. TP expression in human solid tumors occurs in both tumor epithelial cells and in tumor-associated macrophages. The migration assay was adapted to use TP-transfected carcinoma cells to stimulate HUVEC migration, and they were found to induce more migration than did control vector-transfected cells. Human monocyte cells U937 and THP1, which constitutively expressed high levels of TP, also strongly induced HUVEC migration in the coculture assay. CIMU inhibited tumor-cell and monocyte-induced migration. In contrast, a neutralizing antibody to TP had no effect on cell-stimulated HUVEC migration, even though it completely blocked the migration mediated by purified TP. Thus, the intracellular actions of TP were sufficient to stimulate HUVEC chemotaxis. In contrast to purified TP, when incubated with [5'-3H]-thymidine, cells expressing TP released up to 20-fold more 2dR into the medium than dR-1-P. These studies demonstrate that TP-expressing cells mediate EC migration via the intracellular metabolism of thymidine and subsequent extracellular release of 2dR, which forms a chemotactic gradient.


Development of oral side preference during chewing and its relation to hand preference in normal 2- to 8-year-old children.
Normative data on skills of the tongue used in eating are presented. Normal children 5 to 8 years old were studied regarding their preference of placing food either on the right or left side of the mouth when starting to eat. A developmental curve spanning 2 to 8 years was generated by including data from an earlier study. Data of oral side preference were correlated with data of preferred hand use. In addition, the ability to move a small piece of food from one side of the mouth to the other was studied. It was found that normal children undergo a transition from placing solid food predominantly on the right side at 2 years of age to placing it on the left side at 4 years of age. The left side preference persists until at least 8 years of age for both viscous and solid food textures. Oral side preference did not correlate with hand preference. The ability to move food from one side of the mouth to the other (lateralizing) was found to undergo a developmental progression: The inability to lateralize in a third of 2-year-olds gave way to rolling movements. Concomitantly, a consistent increase in slow and then smooth movements was found to occur from 2 to 8 years of age. These data provide the clinician with a normative baseline against which eating-impaired children can be compared.


Phylogenetic Analysis of Geographically Diverse Radopholus similis via rDNA Sequence Reveals a Monomorphic Motif.
The nucleic acid sequences of rDNA ITS1 and the rDNA D2/D3 expansion segment were compared for 57 burrowing nematode isolates collected from Australia, Cameroon, Central America, Cuba, Dominican Republic, Florida, Guadeloupe, Hawaii, Nigeria, Honduras, Indonesia, Ivory Coast, Puerto Rico, South Africa, and Uganda. Of the 57 isolates, 55 were morphologically similar to Radopholus similis and seven were citrus-parasitic. The nucleic acid sequences for PCR-amplified ITS1 and for the D2/D3 expansion segment of the 28S rDNA gene were each identical for all putative R. similis. Sequence divergence for both the ITS1 and the D2/D3 was concordant with morphological differences that distinguish R. similis from other burrowing nematode species. This result substantiates previous observations that the R. similis genome is highly conserved across geographic regions. Autapomorphies that would delimit phylogenetic lineages of non-citrus-parasitic R. similis from those that parasitize citrus were not observed. The data presented herein support the concept that R. similis is comprised of two pathotypes-one that parasitizes citrus and one that does not.


An effective and safe sedation technique combining target-controlled infusion pump with propofol, intravenous pentazocine, and bispectral index monitoring for peroral double-balloon endoscopy.
Because peroral double-balloon endoscopy (DBE) is a time-consuming, painful procedure, sedation with analgesics, and/or anesthetics is generally required. The aim of this prospective study was to investigate the safety and efficacy of our sedation protocol for peroral DBE, which consisted of target-controlled infusion (TCI) anesthesia with propofol, an intravenous bolus of pentazocine, and bispectral index (BIS) monitoring. A total of 34 consecutive patients who underwent DBE by the oral approach were enrolled. Patients were primarily sedated with a continuous infusion of propofol and adjusted in accordance with the BIS levels. The bolus infusion of pentazocine was performed when the propofol infusion was insufficient. The primary outcome measure of this study was to ensure the safety and efficacy of this sedation technique. The secondary purpose was to identify the characteristics of the patient who required the bolus infusion of pentazocine. Five patients (14.7%) required a reduction in the dose of propofol. However, no patient experienced any serious adverse events. All patients (100%) and 80.6% (25/31) of endoscopists answered that the sedation protocol was 'excellent' or 'enough' for peroral DBE. Eleven patients (32.3%) required a bolus injection of pentazocine. Age <60 years and a total procedure time of >70 min were significant risk-factors for pentazocine use. A combination of propofol via TCI pump, bolus injection of pentazocine as needed, and BIS monitoring was a safe and effective procedure for peroral DBE. Reasonable satisfaction indices were obtained from both patients and endoscopists. Pentazocine was required for young patients and in cases with longer procedure times.


Psychometric properties of worry and anxiety measures in a sample of african american and caucasian older adults.
Objective: The aim of our study was to examine the psychometric properties of commonly used anxiety and worry assessment measures in a community-based, low-income sample of African American and Caucasian older adults.Method: African American and Caucasian participants from three community-based clinical trials testing treatments for late-life worry/anxiety were pooled to examine the factor structure, internal consistency reliability, and convergent, discriminant, and predictive validity of the Penn State Worry Questionnaire-Abbreviated (PSWQ-A), Generalized Anxiety Disorder-7 (GAD-7) and Geriatric Anxiety Inventory-short form (GAI-SF).Results: All three measures demonstrated an adequate fit to a one-factor structure. Internal consistency reliability was adequate for the PSWQ-A and GAD-7 in the total sample and racial subgroups but was acceptable for the GAI-SF only in the African American subgroup. The PSWQ-A and GAD-7 demonstrated good convergent, discriminant, and predictive validity. The GAI-SF has adequate convergent and divergent validity in the African American subgroup.Conclusion: Our study offers preliminary evidence for use of the PSWQ-A and GAD-7 for assessment of anxiety in a sample of low-income, predominantly African American participants. These measures may facilitate identification of anxiety symptoms, which are often overlooked in this population. More research is needed to examine the accuracy of these measures in other racial/ethnic groups.


Pinch Loss Ameliorates Obesity, Glucose Intolerance, and Fatty Liver by Modulating Adipocyte Apoptosis in Mice.
The mammalian focal adhesion proteins Pinch1/2 activate integrins and promote cell-extracellular matrix adhesion and migration; however, their roles in adipose tissue and metabolism are unclear. Here we find that high-fat diet (HFD) feeding dramatically increases expression of Pinch1/2 proteins in white adipose tissue (WAT) in mice. Furthermore, expression of Pinch1 is largely upregulated in WAT in leptin-deficient ob/ob type 2 diabetic mice and obese humans. While mice with loss of Pinch1 in adipocytes or global Pinch2 do not display any notable phenotypes, deleting Pinch1 in adipocytes and Pinch2 globally significantly decreases body weight and WAT mass, but not brown adipose tissue mass, in HFD-fed, but not normal chow diet-fed, mice. Pinch loss ameliorates HFD-induced glucose intolerance and fatty liver. After HFD challenge, Pinch loss slightly but significantly accelerates energy expenditure. While Pinch loss decreases adipocyte size and alters adipocyte size distribution, it greatly accelerates cell apoptosis primarily in epididymal WAT and to a lesser extent in subcutaneous WAT. In vitro studies demonstrate that Pinch loss accelerates adipocyte apoptosis by activating the Bim/Caspase-8 pathway. In vivo, genetic ablation of Caspase-8 expression in adipocytes essentially abolishes the ameliorating effects of Pinch deficiency on obesity, glucose intolerance, and fatty liver in mice. Thus, we demonstrate a previously unknown function of Pinch in control of adipose mass, glucose, and fat metabolism via modulation of adipocyte apoptosis. We may define a novel target for the prevention and treatment of metabolic diseases, such as obesity and diabetes.


Does topical Amethocaine cream increase first-time successful cannulation in children compared with a eutectic mixture of local anaesthetics (EMLA) cream? A systematic review and meta-analysis of randomised controlled trials.
Cannulation of children is often required for administration of intravenous fluids and medications, but can cause pain and anxiety. Amethocaine and a eutectic mixture of local anaesthetics (EMLA) cream are two of the most commonly used local anaesthetic creams. To examine the evidence for the superiority of Amethocaine cream compared with EMLA cream in facilitating successful first-time cannulation in children. A systematic search was undertaken in MEDLINE and EMBASE in June 2014. Studies examining cannulation, undertaken with children and providing data about first-time cannulation success rates were considered for inclusion. Three randomised controlled trials met the inclusion criteria and were included in the meta-analysis. Data extraction was undertaken independently by the two authors using predefined data fields. Pooled analysis was based on a random effects model. Low statistical heterogeneity was observed. Amethocaine cream increased the likelihood of successful first-time cannulation (RR 1.046, CI 0.975 to 1.122), although this did not reach statistical significance (p=0.211). Amethocaine cream does not appear to significantly facilitate successful first-time cannulation. Lack of precision and design weaknesses of the included studies hinder the formation of a strong recommendation for either cream. Based on the evidence reviewed here and considering analgesic properties and cost-savings associated with both creams, a weak recommendation can be issued in favour of Amethocaine cream for cannulation in children based on high-quality evidence but where the treatment choice will depend on other factors including cost and provider preference.


Omics for prediction of environmental health effects: Blood leukocyte-based cross-omic profiling reliably predicts diseases associated with tobacco smoking.
The utility of blood-based omic profiles for linking environmental exposures to their potential health effects was evaluated in 649 individuals, drawn from the general population, in relation to tobacco smoking, an exposure with well-characterised health effects. Using disease connectivity analysis, we found that the combination of smoking-modified, genome-wide gene (including miRNA) expression and DNA methylation profiles predicts with remarkable reliability most diseases and conditions independently known to be causally associated with smoking (indicative estimates of sensitivity and positive predictive value 94% and 84%, respectively). Bioinformatics analysis reveals the importance of a small number of smoking-modified, master-regulatory genes and suggest a central role for altered ubiquitination. The smoking-induced gene expression profiles overlap significantly with profiles present in blood cells of patients with lung cancer or coronary heart disease, diseases strongly associated with tobacco smoking. These results provide proof-of-principle support to the suggestion that omic profiling in peripheral blood has the potential of identifying early, disease-related perturbations caused by toxic exposures and may be a useful tool in hazard and risk assessment.


Diet analysis by next-generation sequencing indicates the frequent consumption of introduced plants by the critically endangered red-headed wood pigeon (Columba janthina nitens) in oceanic island habitats.
Oceanic island ecosystems are vulnerable to the introduction of alien species, and they provide a habitat for many endangered species. Knowing the diet of an endangered animal is important for appropriate nature restoration efforts on oceanic islands because introduced species may be a major component of the diets of some endangered species. DNA barcoding techniques together with next-generation sequencing may provide more detailed information on animal diets than other traditional methods. We performed a diet analysis using 48 fecal samples from the critically endangered red-headed wood pigeon that is endemic to the Ogasawara Islands based on chloroplast trnL P6 loop sequences. The frequency of each detected plant taxa was compared with a microhistological analysis of the same sample set. The DNA barcoding approach detected a much larger number of plants than the microhistological analysis. Plants that were difficult to identify by microhistological analysis after being digested in the pigeon stomachs were frequently identified only by DNA barcoding. The results of the barcoding analysis indicated the frequent consumption of introduced species, in addition to several native species, by the red-headed wood pigeon. The rapid eradication of specific introduced species may reduce the food resources available to this endangered bird; thus, balancing eradication efforts with the restoration of native food plants should be considered. Although some technical problems still exist, the trnL approach to next-generation sequencing may contribute to a better understanding of oceanic island ecosystems and their conservation.


Dietary elevated sucrose modulation of diesel-induced genotoxicity in the colon and liver of Big Blue rats.
Earlier studies have indicated that sucrose possesses either co-carcinogenic or tumor-promoter effects in colon carcinogenesis induced by genotoxic carcinogens. In this study we investigated the role of sucrose on diesel exhaust particle (DEP)-induced genotoxicity in the colonic mucosa and liver. Big Blue rats were fed with DEP (0.8 ppm in feed) and/or sucrose (3.45% or 6.85% w/w in feed) for 3 weeks. DEP increased both DNA strand breaks and DNA adducts in colon. Interestingly, sucrose also increased the level of bulky DNA adducts in colon. DEP and sucrose had no effect on DNA strand-breaks and DNA adducts in liver. DEP and sucrose treatment did not have any effect on mutation frequency in colon and liver. Oxidative DNA damage detected as 8-oxodG (8-oxo-7,8-dihydro-2'-deoxyguanosine) and endonuclease III or formamidopyrimidine DNA glycosylase sensitive sites was unaltered in colon and liver. The mRNA expression levels of the DNA repair enzymes N-methylpurine DNA glycosylase ( MPG), 8-oxoguanine DNA glycosylase ( OGG1) and ERCC1 (part of the nucleotide excision repair complex) measured by reverse transcription-polymerase chain reaction were increased in liver by DEP feeding. In colon, expression was unaffected by DEP or sucrose feeding. Among biomarkers of oxidative stress, including vitamin C, malondialdehyde and protein oxidations (gamma-glutamyl semialdehyde and 2-amino adipic semialdehyde) in plasma and liver, only malondialdehyde was increased in plasma by sucrose/DEP feeding. In conclusion, sucrose feeding did not increase DEP-induced DNA damage in colon or liver.


TopBP1 interacts with BLM to maintain genome stability but is dispensable for preventing BLM degradation.
The Bloom syndrome helicase BLM and topoisomerase-IIβ-binding protein 1 (TopBP1) are key regulators of genome stability. It was recently proposed that BLM phosphorylation on Ser338 mediates its interaction with TopBP1, to protect BLM from ubiquitylation and degradation (Wang et al., 2013). Here, we show that the BLM-TopBP1 interaction does not involve Ser338 but instead requires BLM phosphorylation on Ser304. Furthermore, we establish that disrupting this interaction does not markedly affect BLM stability. However, BLM-TopBP1 binding is important for maintaining genome integrity, because in its absence cells display increased sister chromatid exchanges, replication origin firing and chromosomal aberrations. Therefore, the BLM-TopBP1 interaction maintains genome stability not by controlling BLM protein levels, but via another as-yet undetermined mechanism. Finally, we identify critical residues that mediate interactions between TopBP1 and MDC1, and between BLM and TOP3A/RMI1/RMI2. Taken together, our findings provide molecular insights into a key tumor suppressor and genome stability network.


Variability in temporary emigration rates of individually marked female Weddell seals prior to first reproduction.
In many species, temporary emigration (TE) is a phenomenon, often indicative of life-history characteristics such as dormancy, skipped reproduction, or partial migration, whereby certain individuals in a population are temporarily unobservable at a particular site. TE may be a flexible condition-dependent strategy that allows individuals to mitigate effects of adverse conditions. Consequently, TE rates ought to be highly variable, but sources of variations are poorly understood for most species. We used data from known-aged female Weddell seals (Leptonychotes weddellii) tagged in Erebus Bay, Antarctica, to investigate sources of variation in TE rates prior to reproduction and to evaluate possible implications for age-specific probability of first reproduction. TE rates were near 1 the year after birth, decreased to an average of 0.15 (SE = 0.01) by age 8, and were similar thereafter. TE rates varied substantially from year-to-year and were lower for seals that attended reproductive colonies the previous year than for seals that did not attend (e.g., ψ(i,age 8)(UU) - ψ(i, age 8)(PU) = 0.22). Recruitment rates were marginally greater for seals that did attend than for seals that did not attend colonies the previous year. For Weddell seals specifically, our results suggest that (1) motivation to attend colonies varied temporally, (2) as seals grew older they had increased motivation to attend even before reproductive maturity, and (3) seals appear to follow various attendance strategies. More broadly, our results support the idea of TE as a variable, condition-dependent strategy, and highlight the utility of TE models for providing population and life-history insights for diverse taxa.


Mid infrared pulse shaping by optical parametric amplification and its application to optical free induction decay measurement.
We produce microjoule energy shaped mid infrared (MIR) pulses in an optical parametric amplification (OPA) process by imposing the phase and amplitude profile of an arbitrarily shaped pump pulse onto the idler pulse. Using phase locked pulses created using this technique, we measure for the first time, complex optical free induction decay (OFID) of the vibrational coherence of a C-H stretching mode.


HPTLC determination of chemical composition variability in raw materials used in botanicals.
Besides the chemotaxonomic value, nowadays determination of biodiversity and chemical variability has a commercial impact. The exact identity of raw material and constituents of botanical products, such as food supplements or herbal remedies, is a very important argument, being the real prerequisite for quality control and traceability, followed by the determination of active components. However, the analytical approach must consider the natural great variability in secondary metabolites and product form, such as in extracts. Against the reductive approach, on the basis of single chemical standards, so far dominant in Pharmacopoeias monographs, we report applications and utility of the high-performance thin-layer chromatography fingerprint in determination of species of the same genus, of populations of the same species and of different drugs of the same plant.


Blurring emotional memories using eye movements: individual differences and speed of eye movements.
In eye movement desensitization and reprocessing (EMDR), patients make eye movements (EM) while recalling traumatic memories. Making EM taxes working memory (WM), which leaves less resources available for imagery of the memory. This reduces memory vividness and emotionality during future recalls. WM theory predicts that individuals with small working memory capacities (WMCs) benefit more from low levels of taxing (i.e., slow EM) whereas individuals with large WMC benefit more from high levels of taxing (i.e., fast EM). We experimentally examined and tested four prespecified hypotheses regarding the role of WMC and EM speed in reducing emotionality and vividness ratings: 1) EM-regardless of WMC and EM speed-are more effective compared to no dual task, 2) increasing EM speed only affects the decrease in memory ratings irrespective of WMC, 3) low-WMC individuals-compared to high-WMC individuals-benefit more from making either type of EM, 4) the EM intervention is most effective when-as predicted by WM theory-EM are adjusted to WMC. Undergraduates with low (n=31) or high (n=35) WMC recalled three emotional memories and rated vividness and emotionality before and after each condition (recall only, recall + slow EM, and recall + fast EM). Contrary to the theory, the data do not support the hypothesis that EM speed should be adjusted to WMC (hypothesis 4). However, the data show that a dual task in general is more effective in reducing memory ratings than no dual task (hypothesis 1), and that a more cognitively demanding dual task increases the intervention's effectiveness (hypothesis 2). Although adjusting EM speed to an individual's WMC seems a straightforward clinical implication, the data do not show any indication that such a titration is helpful.


Incidence of and risk factors for nosocomial bloodstream infections in adults in the United States, 2003.
Although many studies have examined nosocomial bloodstream infection (BSI), US national estimates of incidence and case-fatality rates have seldom been reported. The purposes of this study were to generate US national estimates of the incidence and severity of nosocomial BSI and to identify risk factors for nosocomial BSI among adults hospitalized in the United States on the basis of a national probability sample. This cross-sectional study used the US Nationwide Inpatient Sample for the year 2003 to estimate the incidence and case-fatality rate associated with nosocomial BSI in the total US population. Cases of nosocomial BSI were defined by using 1 or more International Classification of Diseases, 9th Revision, Clinical Modification codes in the secondary field(s) that corresponded to BSIs that occurred at least 48 hours after admission. The comparison group consisted of all patients without BSI codes in their NIS records. Weighted data were used to generate US national estimates of nosocomial BSIs. Logistic regression was used to identify independent risk factors for nosocomial BSI. The US national estimated incidence of nosocomial BSI was 21.6 cases per 1,000 admissions, while the estimated case-fatality rate was 20.6%. Seven of the 10 leading causes of hospital admissions associated with nosocomial BSI were infection related. We estimate that 541,081 patients would have acquired a nosocomial BSI in 2003, and of these, 111,427 would have died. The final multivariate model consisted of the following risk factors: central venous catheter use (odds ratio [OR], 4.76), other infections (OR, 4.61), receipt of mechanical ventilation (OR, 4.97), trauma (OR, 1.98), hemodialysis (OR, 4.83), and malnutrition (OR, 2.50). The total maximum rescaled R(2) was 0.22. The Nationwide Inpatient Sample was useful for estimating national incidence and case-fatality rates, as well as examining independent predictors of nosocomial BSI.


Chronic inflammation and angiogenic signaling axis impairs differentiation of dental-pulp stem cells.
Dental-pulp tissue is often exposed to inflammatory injury. Sequested growth factors or angiogenic signaling proteins that are released following inflammatory injury play a pivotal role in the formation of reparative dentin. While limited or moderate angiogenesis may be helpful for dental pulp maintenance, the induction of significant level of angiogenesis is probably highly detrimental. Hitherto, several studies have addressed the effects of proinflammatory stimuli on the survival and differentiation of dental-pulp stem cells (DPSC), in vitro. However, the mechanisms communal to the inflammatory and angiogenic signaling involved in DPSC survival and differentiation remain unknown. Our studies observed that short-term exposure to TNF-α (6 and 12 hours [hrs]) induced apoptosis with an upregulation of VEGF expression and NF-κB signaling. However, long-term (chronic) exposure (14 days) to TNF-α resulted in an increased proliferation with a concomitant shortening of the telomere length. Interestingly, DPSC pretreated with Nemo binding domain (NBD) peptide (a cell permeable NF-κB inhibitor) significantly ameliorated TNF-α- and/or VEGF-induced proliferation and the shortening of telomere length. NBD peptide pretreatment significantly improved TNF-α-induced downregulation of proteins essential for differentiation, such as bone morphogenic proteins (BMP)-1 & 2, BMP receptor isoforms-1&2, trasnforming growth factor (TGF), osteoactivin and osteocalcin. Additionally, inhibition of NF-κB signaling markedly increased the mineralization potential, a process abrogated by chronic exposure to TNF-α. Thus, our studies demonstrated that chronic inflammation mediates telomere shortening via NF-κB signaling in human DPSC. Resultant chromosomal instability leads to an emergence of increased proliferation of DPSC, while negatively regulating the differentiation of DPSC, in vitro.


Redundant target effect and intersensory facilitation from visual-tactile interactions in simple reaction time.
In a simple reaction time (RT) task, normal observers responded faster to simultaneous visual and tactile stimuli than to single visual or tactile stimuli. RT to simultaneous visual and tactile stimuli was also faster than RT to simultaneous dual visual or tactile stimuli. The advantage for RT to combined visual-tactile stimuli over RT to the other types of stimulation could be accounted for by intersensory neural facilitation rather than by probability summation. The direction of gaze (and presumably of visual attention) to space regions near to or far from the site of tactile stimulation had no effect on tactile RT. However, RT to single or dual tactile stimuli was fastest when observers could see the sites of tactile stimulation on their hands both directly and through a mirror at the same time. All these effects can be ascribed to the convergence of tactile and visual inputs onto neural centers which contain flexible multimodal representations of body parts.


The long and winding road to useful predictive factors for anti-EGFR therapy in metastatic colorectal carcinoma: the KRAS/BRAF pathway.
Monoclonal antibodies targeting the epidermal growth factor receptor (EGFR) have improved outcomes for patients with metastatic colorectal carcinoma. Among patients not carrying activating mutations in the KRAS gene, only a limited number will experience tumor response to these therapeutic agents. The role of BRAF mutations in determining resistance to this treatment is emerging through preclinical and clinical studies. Standardization and validation of laboratory mutation analysis is needed to allow an optimal use of anti-EGFR therapies in the management of colorectal carcinoma. Clinical single-arm and randomized studies were conducted both in first-line and refractory settings to evaluate the correlation of KRAS mutational status and efficacy of cetuximab and panitumumab. The main trials on first-line regiments are CRYSTAL, which is looking at FOLFIRI (folinic acid, fluorouracil, irinotecan) + cetuximab, and OPUS, which is evaluating FOLFOX (folinic acid, fluorouracil, oxaliplatin) + cetuximab. The results of these trials have induced the European Medicines Agency to apply restrictions to its approval of cetuximab and panitumumab for use in metastatic colorectal cancer patients with wild-type KRAS tumors. However, the absence of KRAS mutations is not sufficient to assure clinical response to cetuximab and panitumumab. We need to discover further molecular biomarkers of impairment in this or other signaling pathways to identify responders more specifically. Preclinical rationale is available for combined therapies, which simultaneously target EGFR and the RAS/RAF/MAPK signaling pathways for metastatic colorectal cancer.


Interactions of the EphA2 Kinase Domain with PIPs in Membranes: Implications for Receptor Function.
EphA2 is a member of the receptor tyrosine kinase family. Interactions of the cytoplasmic region of EphA2 with the cell membrane are functionally important and yet remain incompletely characterized. Molecular dynamics simulations combined with biochemical studies reveal the interactions of the transmembrane, juxtamembrane (JM), and kinase domains with the membrane. We describe how the kinase domain is oriented relative to the membrane and how the JM region can modulate this interaction. We highlight the role of phosphatidylinositol phosphates (PIPs) in mediating the interaction of the kinase domain with the membrane and, conversely, how positively charged patches at the kinase surface and in the JM region induce the formation of nanoclusters of PIP molecules in the membrane. Integration of these results with those from previous studies enable computational reconstitution of a near complete EphA2 receptor within a membrane, suggesting a role for receptor-lipid interactions in modulation of EphA2.


Angiostatin inhibits endothelial MMP-2 and MMP-14 expression: a hypoxia specific mechanism of action.
Angiostatin is an angiogenesis inhibitor in part generated by and released from platelets. Since platelets upon thrombus formation can give rise to areas of hypoxia, we investigated the effects of angiostatin on endothelial cell migration and apoptosis during hypoxia. Human microvascular endothelial cells (HMVEC-L) were exposed to angiostatin under normoxic or hypoxic conditions. Apoptosis was measured by flow-cytometry. HMVEC-L migration was studied using a modified Boyden Chamber assay, in which migration is MMP-dependent. MMP-2, MMP-14, and VEGF levels were measured using immunoblot, Q-PCR and ELISA. During hypoxia HMVEC-L were protected from angiostatin-induced apoptosis due to increased hypoxia-induced VEGF expression. However, MMP-dependent migration of HMVEC-L was inhibited by angiostatin under hypoxic but not normoxic conditions. Angiostatin decreased MMP-2 at the gene and protein levels only in HMVEC-L exposed to hypoxia. A similar result was obtained for MMP-14. Higher angiostatin concentrations, as would be seen during thrombosis, induced HMVEC-L apoptosis, which was not rescued by VEGF. Under hypoxic conditions angiostatin's primary anti-angiogenic mechanism is likely inhibition of endothelial cell MMP-dependent endothelial cell migration. Only at higher concentrations does angiostatin induce endothelial cell death. This study identifies a novel angiostatin anti-angiogenesis mechanism that is only triggered under pathological-like conditions.


Collision tumors: CAMEL, METRO and other acronyms.
We describe two exceptional collision tumors, namely: a 63-year-old woman revealing a melanocytic tumor within a trichoblastoma and a 71-year-old woman with a squamous cell carcinoma colonized by dendritic cells of a melanoma. Both neoplasms showed two different tumor components with intimate relationship. The lesions are labeled in a "playful" way with the acronyms METRO (MElanocytic tumor +TRichOblastoma) and CAMEL (CArcinoma +MELanoma) to facilitate memorization.


Structural and functional comparison of SARS-CoV-2-spike receptor binding domain produced in Pichia pastoris and mammalian cells.
The yeast Pichia pastoris is a cost-effective and easily scalable system for recombinant protein production. In this work we compared the conformation of the receptor binding domain (RBD) from severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) Spike protein expressed in P. pastoris and in the well established HEK-293T mammalian cell system. RBD obtained from both yeast and mammalian cells was properly folded, as indicated by UV-absorption, circular dichroism and tryptophan fluorescence. They also had similar stability, as indicated by temperature-induced unfolding (observed Tm were 50 °C and 52 °C for RBD produced in P. pastoris and HEK-293T cells, respectively). Moreover, the stability of both variants was similarly reduced when the ionic strength was increased, in agreement with a computational analysis predicting that a set of ionic interactions may stabilize RBD structure. Further characterization by high-performance liquid chromatography, size-exclusion chromatography and mass spectrometry revealed a higher heterogeneity of RBD expressed in P. pastoris relative to that produced in HEK-293T cells, which disappeared after enzymatic removal of glycans. The production of RBD in P. pastoris was scaled-up in a bioreactor, with yields above 45 mg/L of 90% pure protein, thus potentially allowing large scale immunizations to produce neutralizing antibodies, as well as the large scale production of serological tests for SARS-CoV-2.


Functional reconstitution into liposomes of purified human RhCG ammonia channel.
Rh glycoproteins (RhAG, RhBG, RhCG) are members of the Amt/Mep/Rh family which facilitate movement of ammonium across plasma membranes. Changes in ammonium transport activity following expression of Rh glycoproteins have been described in different heterologous systems such as yeasts, oocytes and eukaryotic cell lines. However, in these complex systems, a potential contribution of endogenous proteins to this function cannot be excluded. To demonstrate that Rh glycoproteins by themselves transport NH(3), human RhCG was purified to homogeneity and reconstituted into liposomes, giving new insights into its channel functional properties. An HA-tag introduced in the second extracellular loop of RhCG was used to purify to homogeneity the HA-tagged RhCG glycoprotein from detergent-solubilized recombinant HEK293E cells. Electron microscopy analysis of negatively stained purified RhCG-HA revealed, after image processing, homogeneous particles of 9 nm diameter with a trimeric protein structure. Reconstitution was performed with sphingomyelin, phosphatidylcholine and phosphatidic acid lipids in the presence of the C(12)E(8) detergent which was subsequently removed by Biobeads. Control of protein incorporation was carried out by freeze-fracture electron microscopy. Particle density in liposomes was a function of the Lipid/Protein ratio. When compared to empty liposomes, ammonium permeability was increased two and three fold in RhCG-proteoliposomes, depending on the Lipid/Protein ratio (1/300 and 1/150, respectively). This strong NH(3) transport was reversibly inhibited by mercuric and copper salts and exhibited a low Arrhenius activation energy. This study allowed the determination of ammonia permeability per RhCG monomer, showing that the apparent Punit(NH3) (around 1x10(-3) microm(3)xs(-1)) is close to the permeability measured in HEK293E cells expressing a recombinant human RhCG (1.60x10(-3) microm(3)xs(-1)), and in human red blood cells endogenously expressing RhAG (2.18x10(-3) microm(3)xs(-1)). The major finding of this study is that RhCG protein is active as an NH(3) channel and that this function does not require any protein partner.


A General Model of the Relationship between the Apportionment of Human Genetic Diversity and the Apportionment of Human Phenotypic Diversity.
Models that examine genetic differences between populations alongside a genotype-phenotype map can provide insight about phenotypic variation among groups. We generalize a simple model of a completely heritable, additive, selectively neutral quantitative trait to examine the relationship between single-locus genetic differentiation and phenotypic differentiation on quantitative traits. In agreement with similar efforts using different models, we show that the expected degree to which two groups differ on a neutral quantitative trait is not strongly affected by the number of genetic loci that influence the trait: neutral trait differences are expected to have a magnitude comparable to the genetic differences at a single neutral locus. We discuss this result with respect to population differences in disease phenotypes, arguing that although neutral genetic differences between populations can contribute to specific differences between populations in health outcomes, systematic patterns of difference that run in the same direction for many genetically independent health conditions are unlikely to be explained by neutral genetic differentiation.


Expansive generation of functional airway epithelium from human embryonic stem cells.
Production of human embryonic stem cell (hESC)-derived lung progenitors has broad applicability for drug screening and cell therapy; however, this is complicated by limitations in demarcating phenotypic changes with functional validation of airway cell types. In this paper, we reveal the potential of hESCs to produce multipotent lung progenitors using a combined growth factor and physical culture approach, guided by the use of novel markers LIFRα and NRP1. Lung specification of hESCs was achieved by priming differentiation via matrix-specific support, followed by air-liquid interface to allow generation of lung progenitors capable of in vitro maturation into airway epithelial cell types, resulting in functional characteristics such as secretion of pulmonary surfactant, ciliation, polarization, and acquisition of innate immune activity. This approach provided a robust expansion of lung progenitors, allowing in vivo assessment, which demonstrated that only fully differentiated hESC-derived airway cells were retained in the distal airway, where they aided in physiological recovery in immunocompromised mice receiving airway injury. Our study provides a basis for translational applications of hESCs for lung diseases.


Covert hepatic encephalopathy leads to distinct alterations in the emotional state, independently of MELD-Score.
Covert hepatic encephalopathy impairs many aspects of quality of life, although its impact on the emotional state has not been evaluated. This study aims to evaluate the impact of covert hepatic encephalopathy on the emotional state and which factors are associated with changes in the emotional state in patients with cirrhosis. This single-center study included all patients with cirrhosis who underwent the portosystemic encephalopathy syndrome (PSE) test, critical flicker frequency, and emotional state assessment with the Eigenschaftswörterliste 60-S in 2011. Covert hepatic encephalopathy was defined by abnormal PSE. Parametric and non-parametric tests were used according to variable distribution. One hundred seventeen patients with cirrhosis were included (median age: 59 [interquartile range: 48 - 67], 32 % female, 74 % alcohol-associated). Seventy patients had covert hepatic encephalopathy (60 %) with a higher MELD (16 [interquartile range: 13 - 21], p = 0.001) and a higher Child-Pugh score (p = 0.003) compared to patients without encephalopathy. Patients with covert encephalopathy felt reduced mental activity (p = 0.004), lower general well-being (p = 0.001), and reduced extraversion (p = 0.021). The scores in the negative domains such as general lethargy (p = 0.031) and anxiousness/depressiveness (p = 0.033) were higher in patients with covert hepatic encephalopathy. There was no correlation between MELD and the emotional state. Patients with 2 pathological tests (critical flicker frequency and PSE) showed the most distinct alterations in the emotional state in the group of patients with covert hepatic encephalopathy. Patients with covert hepatic encephalopathy have an alteration of the emotional state, which is more marked in patients with 2 pathological tests. Interestingly, MELD had no impact on the emotional state.


Geochemical characterization of acid mine lakes in northwest Turkey and their effect on the environment.
Mining activity generates a large quantity of mine waste. The potential hazard of mine waste depends on the host mineral. The tendency of mine waste to produce acid mine drainage (AMD) containing potentially toxic metals depends on the amounts of sulfide, carbonate minerals, and trace-element concentrations found in ore deposits. The acid mine process is one of the most significant environmental challenges and a major source of water pollution worldwide. AMD and its effects were studied in northwest Turkey where there are several sedimentary and hydrothermal mineral deposits that have been economically extracted. The study area is located in Can county of Canakkale province. Canakkale contains marine, lagoon, and lake sediments precipitated with volcanoclastics that occurred as a result of volcanism, which was active during various periods from the Upper Eocene to Plio-Quaternary. Can county is rich in coal with a total lignite reserve >100 million tons and contains numerous mines that were operated by private companies and later abandoned without any remediation. As a result, human intervention in the natural structure and topography has resulted in large open pits and deterioration in these areas. Abandoned open pit mines typically fill with water from runoff and groundwater discharge, producing artificial lakes. Acid drainage waters from these mines have resulted in the degradation of surface-water quality around Can County. The average pH and electrical conductivity of acid mine lakes (AMLs) in this study were found to be 3.03 and 3831.33 μS cm(-1), respectively. Total iron (Fe) and aluminum (Al) levels were also found to be high (329.77 and 360.67 mg L(-1), respectively). The results show that the concentration of most elements, such as Fe and Al in particular, exceed national and international water-quality standards.


Monoacyl phosphatidylcholine inhibits the formation of lipid multilamellar structures during in vitro lipolysis of self-emulsifying drug delivery systems.
The colloidal structures formed during lipolysis of self-emulsifying drug delivery systems (SEDDS) might affect the solubilisation and possibly the absorption of drugs. The aim of the current study is to elucidate the structures formed during the in vitro lipolysis of four SEDDS containing medium-chain glycerides and caprylocaproyl polyoxyl-8 glycerides (Labrasol), with or without monoacyl phosphatidylcholine (MAPC). In situ synchrotron small-angle X-ray scattering (SAXS) was combined with ex situ cryogenic transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS) to elucidate the generated structures. The SAXS scattering curves obtained during the lipolysis of MAPC-free SEDDS containing 43-60% w/w Labrasol displayed a lamellar phase peak at q=2.13nm-1 that increased with Labrasol concentration, suggesting the presence of multilamellar structures (MLS) with a d-spacing of 2.95nm. However, SEDDS containing 20-30% w/w MAPC did not form MLS during the lipolysis. The cryo-TEM and DLS studies showed that MAPC-free SEDDS formed coarse emulsions while MAPC-containing SEDDS formed nanoemulsions during the dispersion in digestion medium. From the first minute and during the entire lipolysis process, SEDDS both with and without MAPC generated uni-, bi-, and oligo-lamellar vesicles. The lipolysis kinetics in the first minutes of the four SEDDS correlated with an increased intensity of the SAXS curves and the rapid transformation from lipid droplets to vesicles observed by cryo-TEM. In conclusion, the study elucidates the structures formed during in vitro lipolysis of SEDDS and the inhibitory effect of MAPC on the formation of MLS.


Confined photo-release of nitric oxide with simultaneous two-photon fluorescence tracking in a cellular system.
Nitric oxide (NO) is a key signaling molecule in biological systems. New tools are required to therapeutically modulate NO levels with confined precision. This study explores the photoactivatable properties of an NO releasing compound (CPA), based on cupferron O-alkylated with an anthracene derivative. Upon light stimulation, CPA uncages two species: cupferron, which liberates NO, and an anthrylmethyl carbocation, which evolves into a fluorescent reporter. Proof-of-principle is demonstrated using one- and two-photon excitation (1PE and 2PE) in a cellular system (A431 cells). It was found that 1PE induces cell toxicity, while 2PE does not. Since 1PE using UV light is more likely to generate cellular photodamage, the cell toxicity observed using 1PE is most likely a combinatory effect of NO release and other UV-induced damage, which should be subject to further investigation. On the other hand, absence of phototoxicity using 2PE suggests that NO alone is not cytotoxic. This leads to the conclusion that the concept of 2PE photorelease of NO from CPA enable opportunities for biological studies of NO signaling with confined precision of NO release with minimal cytotoxicity.


A transcriptome derived female-specific marker from the invasive Western mosquitofish (Gambusia affinis).
Sex-specific markers are a prerequisite for understanding reproductive biology, genetic factors involved in sex differences, mechanisms of sex determination, and ultimately the evolution of sex chromosomes. The Western mosquitofish, Gambusia affinis, may be considered a model species for sex-chromosome evolution, as it displays female heterogamety (ZW/ZZ), and is also ecologically interesting as a worldwide invasive species. Here, de novo RNA-sequencing on the gonads of sexually mature G. affinis was used to identify contigs that were highly transcribed in females but not in males (i.e., transcripts with ovary-specific expression). Subsequently, 129 primer pairs spanning 79 contigs were tested by PCR to identify sex-specific transcripts. Of those primer pairs, one female-specific DNA marker was identified, Sanger sequenced and subsequently validated in 115 fish. Sequence analyses revealed a high similarity between the identified sex-specific marker and the 3´ UTR of the aminomethyl transferase (amt) gene of the closely related platyfish (Xiphophorus maculatus). This is the first time that RNA-seq has been used to successfully characterize a sex-specific marker in a fish species in the absence of a genome map. Additionally, the identified sex-specific marker represents one of only a handful of such markers in fishes.


Cannabinoids as gastrointestinal anti-inflammatory drugs.
In this mini-review, we focus on the potential of the endocannabinoid system as a target for novel therapies to treat gastrointestinal (GI) inflammation. We discuss the organization of the endocannabinoid signaling and present possible pharmacological sites in the endocannabinoid system. We also refer to recent clinical findings in the field. Finally, we point at the potential use of cannabinoids at low, non-psychoactive doses to counteract non-inflammatory pathological events in the GI tract, like chemotherapy-induced diarrhea, as evidenced by Abalo et al. in the rat model.


In vitro and in vivo release of nerve growth factor from biodegradable poly-lactic-co-glycolic-acid microspheres.
Regeneration of peripheral nerves after injury is suboptimal. We now report the long term delivery of nerve growth factor (NGF) by biodegradable poly-lactic-co-glycolic acid (PLGA) microspheres in vitro and in vivo. Lactic to glycolic acid ratios of 50:50 and 85:15 were fabricated using the double emulsion solvent, evaporation technique. Three different inherent viscosities (0.1 dL g(-1) : 1A, 0.4 dL g(-1) : 4A, 0.7 dL g(-1) : 7A) were analyzed. In vitro, release of NGF for 23 days was measured. Electron microscopy demonstrated intact spheres for at least 7 days (50:50 1A), 14 days (50:50 4A), or 35 days (50:50 7A and 85:15 7A). In vitro release kinetics was characterized by burst release, followed by release of NGF at a rate of 0.6-1.6% a day. Release curves for 50:50 1A and 85:15 7A differed significantly from other compositions (p < 0.01). In vivo, release was characterized by a novel radionuclide tracking assay. Release rates varied from 0.9 to 2.2% per day with linear kinetics. All but the 85:15 type of spheres showed different release profiles in vivo compared to in vitro conditions. On the basis of the surface morphology and release profiles, we found microspheres fabricated from 50:50 4A PLGA to be best suited for the use in a rat sciatic nerve injury model.


How neuronal migration contributes to the morphogenesis of the CNS: insights from the zebrafish.
We used transgenic zebrafish expressing GFP or YFP in subpopulations of neurons to study the migration, homing process and axon extension of groups of CNS neurons in different regions of the zebrafish brain. We found that extensive migration takes place at all levels of the CNS and gives rise to nuclei or cell populations with specific identities. Here, we describe 4 previously unknown or only partially characterized migratory events taking place in the zebrafish telencephalon and rhombic lip, using 3 different transgenic lines, and identify the phenotypes of the cells undertaking these migrations. The migration of a subgroup of mitral cell precursors from the dorsocaudal telencephalon to the olfactory bulb, visualized in the tg(tbr1:YFP) transgenic line, is coupled with morphogenetic transformation of the dorsal telencephalon. The tg(1.4dlx5a-6a:GFP) transgenic line provides a means to analyze the migration of GABAergic interneurons from the ventral to the dorsal telencephalon, thus extending the occurrence of this migration to another vertebrate. The tg(Xeom:GFP) transgenic line provides the first demonstration of the dorsoventral migration of glutamatergic septal neurons, present in mammals and now described in fish, thus reconciling the contrasting evidence of dorsal patterning genes (tbr1, eomes) expressed in a ventral cell population. Furthermore, migration studies in the tg(1.4dlx5a-6a:GFP) and tg(Xeom:GFP) lines help determine the origin of 2 important cell populations in the fish cerebellum: projection neurons and Purkinje cells. These examples reinforce the concept that migratory events contribute to the distribution of cell types with diverse identities through the CNS and that zebrafish transgenic lines represent excellent tools to study these events.


An approximate and efficient method for optimal rotation alignment of 3D models.
In many shape analysis applications, the ability to find the best rotation that aligns two models is an essential first step in the analysis process. In the past, methods for model alignment have either used normalization techniques, such as PCA alignment, or have performed an exhaustive search over the space of rotation to find the best optimal alignment. While normalization techniques have the advantage of efficiency, providing a quick method for registering two shapes, they are often imprecise and can give rise to poor alignments. Conversely, exhaustive search is guaranteed to provide the correct answer, but, even using efficient signal processing techniques, this type of approach can be prohibitively slow. In this paper, we present a new method for aligning two 3D shapes. We show that the method is markedly faster than existing approaches based on efficient signal processing and we provide registration results demonstrating that the alignments obtained using our method have a high degree of precision and are markedly better than those obtained using normalization.


Understanding the infrared spectrum of bare CH5+.
Protonated methane, CH5+, continues to elude definitive structural assignment, as large-amplitude vibrations and hydrogen scrambling challenge both theory and experiment. Here, the infrared spectrum of bare CH5+ is presented, as detected by reaction with carbon dioxide gas after resonant excitation by the free electron laser at the FELIX facility in the Netherlands. Comparison of the experimental spectrum at approximately 110 kelvin to finite-temperature infrared spectra, calculated by ab initio molecular dynamics, supports fluxionality of bare CH5+ under experimental conditions and provides a dynamical mechanism for exchange of hydrogens between CH3 tripod positions and the three-center bonded H2 moiety, which eventually leads to full hydrogen scrambling. The possibility of artificially freezing out scrambling and internal rotation in the simulations allowed assignment of the infrared spectrum despite this pronounced fluxionality.


Transposon-insertion sequencing screens unveil requirements for EHEC growth and intestinal colonization.
Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important food-borne pathogen that colonizes the colon. Transposon-insertion sequencing (TIS) was used to identify genes required for EHEC and E. coli K-12 growth in vitro and for EHEC growth in vivo in the infant rabbit colon. Surprisingly, many conserved loci contribute to EHEC's but not to K-12's growth in vitro. There was a restrictive bottleneck for EHEC colonization of the rabbit colon, which complicated identification of EHEC genes facilitating growth in vivo. Both a refined version of an existing analytic framework as well as PCA-based analysis were used to compensate for the effects of the infection bottleneck. These analyses confirmed that the EHEC LEE-encoded type III secretion apparatus is required for growth in vivo and revealed that only a few effectors are critical for in vivo fitness. Over 200 mutants not previously associated with EHEC survival/growth in vivo also appeared attenuated in vivo, and a subset of these putative in vivo fitness factors were validated. Some were found to contribute to efficient type-three secretion while others, including tatABC, oxyR, envC, acrAB, and cvpA, promote EHEC resistance to host-derived stresses. cvpA is also required for intestinal growth of several other enteric pathogens, and proved to be required for EHEC, Vibrio cholerae and Vibrio parahaemolyticus resistance to the bile salt deoxycholate, highlighting the important role of this previously uncharacterized protein in pathogen survival. Collectively, our findings provide a comprehensive framework for understanding EHEC growth in the intestine.


[Neurourological characterization of myelomeningocele in the adult patient].
To characterize the neurourological features of the vesicourethral dysfunction of patients with cogenital myelomeningocele when they reach the adult age. We report the results of the neurourological physical examination and video urodynamic tests of 52 adult patients born with myelomeningocele, with ages ranging from 18 to 14 years (mean age 20.7) controlled over the last 14 years in the Urodynamics Unit of the Hospital Gregorio Morañon in Madrid. The neurological level of the lesion was evaluated in all of them. Following these results we established the diagnosis of the lesion of the three constitutive elements of the vesicourethral innervation (sympathetic, parasympathetic, and pudendal) in order to classify the lower urinary tract neurogenic dysfunction of the patients based on lesion level and type of damaged innervation. The alteration of the perineal sensitivity and diminishment of the anal sphincter tone, in addition to absence of anal voluntary controlled and bulbocavernosus reflex, where the most frequent findings in the neurourological physical examination. Detrusor behaviour was predominantly areflexic (88.4%), in association with diminished bladder compliance in half of the cases. The sphincter was not competent during the filling phase in 51.9% of the cases, competent in 42.3%, and normal in the remainder 5.7%. During the voiding phase we confirmed the absence of sphincter relaxation in 82.6% of the cases. There was a predominance of the lower level of lesion in all three sympathetic, parasympathetic and pudendal nerves. In almost all patients (96.7%) there was lesion in at least two of the three types of innervation. The most frequent neurogenic dysfunction of the lower urinary tract was the multiple inferior motor neuron pure lesion (65.3%), also finding pure superior motor neuron (3.8%), mixed multiple (9.6%), and pure single lesions (1.9%). Three patients presented a normal vesicourethral function. We demonstrated the multiplicity of lower urinary tract neurogenic dysfunction in adult patients with congenital myelomeningocele, its occasional absence, the lack of correlation with the neurological level of the lesion, and the predominance of the multiple inferior motor neuron pure type of lesion.


Effects of cholinesterase inhibiting sage (Salvia officinalis) on mood, anxiety and performance on a psychological stressor battery.
Salvia officinalis (sage) has previously been shown both to possess in vitro cholinesterase inhibiting properties, and to enhance mnemonic performance and improve mood in healthy young participants. In this double-blind, placebo-controlled, crossover study, 30 healthy participants attended the laboratory on three separate days, 7 days apart, receiving a different treatment in counterbalanced order on each occasion (placebo, 300, 600 mg dried sage leaf). On each day mood was assessed predose and at 1 and 4 h postdose. Each mood assessment comprised completion of Bond-Lader mood scales and the State Trait Anxiety Inventory (STAI) before and after 20 min performance of the Defined Intensity Stress Simulator (DISS) computerized multitasking battery. In a concomitant investigation, an extract of the sage leaf exhibited dose-dependent, in vitro inhibition of acetylcholinesterase and, to a greater extent, butyrylcholinesterase. Both doses of sage led to improved ratings of mood in the absence of the stressor (that is, in pre-DISS mood scores) postdose, with the lower dose reducing anxiety and the higher dose increasing 'alertness', 'calmness' and 'contentedness' on the Bond-Lader mood scales. The reduced anxiety effect following the lower dose was, however, abolished by performing the DISS, with the same dose also being associated with a reduction of alertness during performance. Task performance on the DISS battery was improved for the higher dose at both postdose sessions, but reduced for the lower dose at the later testing session. The results confirm previous observations of the cholinesterase inhibiting properties of S. officinalis, and improved mood and cognitive performance following the administration of single doses to healthy young participants.


Postsynaptic receptor trafficking underlying a form of associative learning.
To elucidate molecular, cellular, and circuit changes that occur in the brain during learning, we investigated the role of a glutamate receptor subtype in fear conditioning. In this form of learning, animals associate two stimuli, such as a tone and a shock. Here we report that fear conditioning drives AMPA-type glutamate receptors into the synapse of a large fraction of postsynaptic neurons in the lateral amygdala, a brain structure essential for this learning process. Furthermore, memory was reduced if AMPA receptor synaptic incorporation was blocked in as few as 10 to 20% of lateral amygdala neurons. Thus, the encoding of memories in the lateral amygdala is mediated by AMPA receptor trafficking, is widely distributed, and displays little redundancy.


Physician-scientist, heal thyself . . .
Historically, physician-scientists have had dual roles in caring for patients and in performing investigative research that could potentially lead to new diagnostics and therapeutics. Physician-scientists conducted teaching rounds in the hospital, surrounded by eager house staff and medical students, and were often avidly pursued as the most important sources of new knowledge for trainees. But alas, times have changed. Now physician-scientists are rarely seen in the hospital; they are most often spotted at their desks tapping out yet another grant application. Most struggle to find the time to mentor students and clinical trainees, let alone to care for patients in the hospital, even though these interactions are often the motivating forces for scientific creativity.


Cerebral amyloid angiopathy in dementia and old age.
A necropsy study of 159 elderly patients drawn mainly from a prospectively assessed geriatric hospital population was carried out to investigate the relationship of cerebral amyloid angiopathy to Alzheimer's disease, other CNS disease and ageing. About half the patients were demented and the majority of these had Alzheimer's disease. In Alzheimer's disease there was an incidence of cerebral amyloid angiopathy of 82%. Among the other groups of patients, both demented and non-demented, the incidence of cerebral amyloid angiopathy was a little over 30%, and remained constant between 60 and 102 years of age.


Hair growth promoting activity of Eclipta alba in male albino rats.
Alopecia is a dermatological disorder with psychosocial implications on patients with hair loss. Eclipta alba Hassk. is a well-known Ayurvedic herb with purported claims of hair growth promotion. In the reported work attempts were undertaken to evaluate petroleum ether and ethanol extract of E. alba Hassk. for their effect on promoting hair growth in albino rats. The extracts were incorporated into oleaginous cream (water in oil cream base) and applied topically on shaved denuded skin of albino rats. The time (in days) required for hair growth initiation as well as completion of hair growth cycle was recorded. Minoxidil 2% solution was applied topically and served as positive control for comparison. Hair growth initiation time was significantly reduced to half on treatment with the extracts, as compared to control animals. The time required for complete hair growth was also significantly reduced. Quantitative analysis of hair growth after treatment with petroleum ether extract (5%) exhibited greater number of hair follicles in anagenic phase (69 +/- 4) which were higher as compared to control (47 +/- 13). The result of treatment with 2 and 5% petroleum ether extracts were better than the positive control minoxidil 2% treatment.


Steps to Enhance Early Recovery After Hematopoietic Stem Cell Transplantation: Lessons Learned From a Physical Activity Feasibility Study.
This pilot study tested and refined a free-living physical activity intervention. The investigators evaluated the acceptability and feasibility of the intervention after hematopoietic stem cell transplantation and determined preliminary effects on physical activity, fatigue, muscle strength, functional ability, and quality of life. This pilot study used a 1-group, pretest-posttest design. The free-living physical activity intervention consisted of an education component and 6 weeks of gradually increasing physical activity after discharge from the hospital. The intervention was designed to increase steps by 10% weekly. Subjects were assessed before transplantation and during the seventh week after discharge from the hospital after completing the intervention. Pretest-posttest scores were analyzed with paired t tests. Subject wore the physical activity tracker for an average of 38 of 42 days and met their physical activity goals 57% of the time. Subjects reported significantly less physical fatigue after the free-living physical activity intervention compared with baseline (P = .05). Improvements in quality of life approached significance (P = .06). The findings demonstrate that the free-living physical activity intervention implemented during the very early recovery period after transplantation is feasible and acceptable. The intervention potentially reduces fatigue and improves quality of life. The positive results must be interpreted cautiously given the pilot nature of the study. The evidence supports continued investigation.


Theory and delivery of health programming in the community: the Pawtucket Heart Health Program.
The Pawtucket Heart Health Program is one of the community studies examining whether population-based efforts to lower cardiovascular risk factors will reduce cardiovascular morbidity and mortality. The Pawtucket Heart Health Program intervention is based on a blend of social learning theory, community organization models, community psychology tenets, and diffusion research. This model allows for multifaceted programs that target individuals, groups, organizations, and the entire community to alter their cardiovascular risk through managing blood pressure, lowering blood cholesterol, quitting smoking, increasing fitness, and maintaining desirable weight levels. A dominant feature of the intervention is the emphasis that it places on volunteers for program delivery. The role of volunteers in providing direct services to help citizens lower their blood pressure and lose weight is highlighted to demonstrate the feasibility and effectiveness of these services. In addition, church-based programming which utilizes volunteers to manage and direct programs is also presented as an example of community-based health promotion efforts that promote collective efficacy.


Polystomatidae (Monogenea) of African Anura: Polystoma dawiekoki n. sp. parasitic in Ptychadena anchietae (Bocage).
Polystoma dawiekoki n. sp. is described as a new species of the Polystomatidae parasitic in the urinary bladder of the plain grass frog Ptychadena anchietae. This parasite was collected at Mkuze town and Mkuze Game Reserve in northern Kwazulu-Natal Province, in the Kruger National Park in Mpumalanga Province, South Africa, and at Bulwa in Tanga Province, East Usambara Mountains, Tanzania. It is distinguished from other African Polystoma species by a combination of characters, including the body size, size and shape of marginal hooklets and the haptor length to body length ratio. The presence of adult, as well as subadult, parasites in the same individuals, as is known for Eupolystoma, represents a significant evolutionary departure from the pattern of transmission typical of Polystoma in most of the other anuran hosts.


Strong regulation of slow anion channels and abscisic acid signaling in guard cells by phosphorylation and dephosphorylation events.
Recent evidence suggests that slow anion channels in guard cells need to be activated to trigger stomatal closing and efficiently inactivated during stomatal opening. The patch-clamp technique was employed here to determine mechanisms that produce strong regulation of slow anion channels in guard cells. MgATP in guard cells, serving as a donor for phosphorylation, leads to strong activation of slow anion channels. Slow anion-channel activity was almost completely abolished by removal of cytosolic ATP or by the kinase inhibitors K-252a and H7. Nonhydrolyzable ATP, GTP, and guanosine 5'-[gamma-thio]triphosphate did not replace the ATP requirement for anion-channel activation. In addition, down-regulation of slow anion channels by ATP removal was inhibited by the phosphatase inhibitor okadaic acid. Stomatal closures in leaves induced by the plant hormone abscisic acid (ABA) and malate were abolished by kinase inhibitors and/or enhanced by okadaic acid. These data suggest that ABA signal transduction may proceed by activation of protein kinases and inhibition of an okadaic acid-sensitive phosphatase. This modulation of ABA-induced stomatal closing correlated to the large dynamic range for up- and down-regulation of slow anion channels by opposing phosphorylation and dephosphorylation events in guard cells. The presented opposing regulation by kinase and phosphatase modulators could provide important mechanisms for signal transduction by ABA and other stimuli during stomatal movements.


Neural regions that underlie reinforcement learning are also active for social expectancy violations.
Prediction error, the difference between an expected and an actual outcome, serves as a learning signal that interacts with reward and punishment value to direct future behavior during reinforcement learning. We hypothesized that similar learning and valuation signals may underlie social expectancy violations. Here, we explore the neural correlates of social expectancy violation signals along the universal person-perception dimensions trait warmth and competence. In this context, social learning may result from expectancy violations that occur when a target is inconsistent with an a priori schema. Expectancy violation may activate neural regions normally implicated in prediction error and valuation during appetitive and aversive conditioning. Using fMRI, we first gave perceivers high warmth or competence behavioral information that led to dispositional or situational attributions for the behavior. Participants then saw pictures of people responsible for the behavior; they represented social groups either inconsistent (rated low on either warmth or competence) or consistent (rated high on either warmth or competence) with the behavior information. Warmth and competence expectancy violations activate striatal regions that represent evaluative and prediction error signals. Social cognition regions underlie consistent expectations. These findings suggest that regions underlying reinforcement learning may work in concert with social cognition regions in warmth and competence social expectancy. This study illustrates the neural overlap between neuroeconomics and social neuroscience.


Paraneoplastic pemphigus is associated with the DRB1*03 allele.
Pemphigus is a group of autoimmune blistering diseases caused by autoantibodies directed against keratinocyte adhesion molecules. Pemphigus vulgaris (PV) and pemphigus foliaceus (PF), in which autoantibodies bind, respectively, to desmoglein 3 and desmoglein 1, are strongly associated with HLA-class II DR4 and DR14 alleles. In paraneoplastic pemphigus (PNP), a rare variant associated with neoplasia, autoantibodies target proteins of the plakin family in addition to desmogleins 1 and 3. The presence of anti-desmoglein antibodies in all types of pemphigus raises the question of common molecular mechanisms of susceptibility, particularly similar MHC-class II allele associations, in the different forms of the disease. HLA-DRB1 typing was performed in 13 PNP patients and results were compared to those obtained from 84 healthy controls, 37 PV and 31 PF patients. Our data demonstrate a significant association of PNP with HLA-DRB1*03 allele which was found in 61.5% of the patients, whereas DRB1*04 and DRB1*14 appear not to be involved in PNP susceptibility. Therefore, the HLA-genetic background of PNP differs from that of other types of pemphigus, which suggests that distinct mechanism(s) initiate(s) the immunological response in this form of pemphigus.


Sudden arrhythmia death syndrome: importance of the long QT syndrome.
In approximately 5 percent of sudden cardiac deaths, no demonstrable anatomic abnormality is found. Some cases are caused by sudden arrhythmia death syndrome. A prolonged QT interval is a common thread among the various entities associated with sudden arrhythmia death syndrome. A number of drugs are known to cause QT prolongation (e.g., terfenadine), as are hypokalemia, hypomagnesemia, myocarditis, and endocrine and nutritional disorders. Recently, attention has focused on a group of inherited gene mutations in cardiac ion channels that cause long QT syndrome and carry an increased risk for sudden death. Some of the highest rates of inherited long QT syndrome occur in Southeast Asian and Pacific Rim countries. The median age of persons who die of long QT syndrome is 32 years; men are predominately affected. In addition to a prolonged QT interval, which occurs in some but not all persons with long QT syndrome, another characteristic electrocardiographic abnormality is the so-called Brugada sign (an upward deflection of the terminal portion of the QRS complex). Most cardiac events are precipitated by vigorous exercise or emotional stress, but they also can occur during sleep. Torsades de pointes and ventricular fibrillation are the usual fatal arrhythmias. Long QT syndrome should be suspected in patients with recurrent syncope during exertion and those with family histories of sudden, unexpected death. Unfortunately, not all persons with long QT syndrome have premonitory symptoms or identifiable electrocardiographic abnormalities, and they may first present with sudden death. Beta blockers, potassium supplements, and implantable defibrillators have been used for treatment of long QT syndrome. Identifying the specific gene mutation in a given patient with long QT syndrome can help guide prophylactic therapy.


A vector set for systematic metabolic engineering in Saccharomyces cerevisiae.
A set of shuttle vectors was constructed to facilitate expression of genes for metabolic engineering in Saccharomyces cerevisiae. Selectable markers include the URA3, TRP1, MET15, LEU2-d8, HIS3 and CAN1 genes. Differential expression of genes can be achieved as each marker is available on both CEN/ARS- and 2 µ-containing plasmids. Unique restriction sites downstream of TEF1, PGK1 or HXT7-391 promoters and upstream of the CYC1 terminator allow insertion of open-reading frame cassettes for expression. Furthermore, a fragment appropriate for integration into the genome via homologous recombination can be readily generated in a polymerase chain reaction. Vector marker genes are flanked by loxP recognition sites for the CreA recombinase to allow efficient site-specific marker deletion and recycling. Expression and copy number were characterized for representative high- and low-copy vectors carrying the different marker and promoter sequences. Metabolic engineering typically requires the stable introduction of multiple genes and genomic integration is often preferred. This requires an expanded number of stable expression sites relative to standard gene expression studies. This study demonstrated the practicality of polymerase chain reaction amplification of an expression cassette and genetic marker, and subsequent replacement of endogenous retrotransposons by homologous recombination with flanking sequences. Such reporters were expressed comparably to those inserted at standard integration loci. This expands the number of available characterized integration sites and demonstrates that such sites provide a virtually inexhaustible pool of integration targets for stable expression of multiple genes. Together these vectors and expression loci will facilitate combinatorial gene expression for metabolic engineering.


Two-dimensional infrared spectroscopy: an emerging analytical tool?
Ultrafast two-dimensional infrared (2D-IR) spectroscopy has provided valuable insights into biomolecular structure and dynamics, but recent progress in laser technology and data analysis methods have demonstrated the potential for high throughput 2D-IR measurements and analytical applications. Using 2D-IR as an analytical tool requires a different approach to data collection and analysis compared to pure research applications however and, in this review, we highlight progress towards usage of 2D-IR spectroscopy in areas relevant to biomedical, pharmaceutical and analytical molecular science. We summarise the technical and methodological advances made to date and discuss the challenges that still face 2D-IR spectroscopy as it attempts to transition from the state-of-the-art laser laboratory to the standard suite of analytical tools.


Antiviral activity of theaflavin digallate against herpes simplex virus type 1.
Tea is the second most consumed drink in the world. The beneficial effects of tea have been mostly attributed to its catechin content. Black tea is derived from the leaves of Camellia sinensis plant, and it is rich in theaflavin polyphenols, in particular theaflavin (TF1), theaflavin-3-monogallate (TF2A), theaflavin-3'-monogallate (TF2B), and theaflavin-3,3'-digallate (TF3). Vero and A549 cells were used to evaluate the effect of purified individual black tea theaflavins as anti-herpes simplex virus 1 agents. With the rise of HSV resistant strains, there is a critical need to develop novel antiherpesviral treatments. Results of the cytotoxicity assay tested by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium] showed that TF1, TF2, and TF3 are not toxic to Vero and A549 cells at a concentration up to 75 μM. The antiviral activity of the individual theaflavins was tested by plaque reduction assay, MTS assay, flow cytometric analysis and confocal microscopy observations. The results showed that TF1, TF2, and TF3 exhibit potent, dose-dependent anti-HSV-1 effect, with TF3 being the most efficient in both Vero and A549 cells. A concentration of 50 μM TF3 and above was sufficient to inhibit >99% of the production of HSV-1 viral particles. The anti-HSV-1 effect of TF3 is due to a direct effect on the virions, and treating Vero or A549 cells with TF3 for 1h prior to infection, or treating the cells at different times post infection does not inhibit HSV-1 production. TF3 is stable at vaginal pH, indicating its potential to be a promising natural and affordable remedy against herpes simplex viral infections.


Rad4-Rad23 interaction with SWI/SNF links ATP-dependent chromatin remodeling with nucleotide excision repair.
Chromatin rearrangement occurs during nucleotide excision repair (NER). Here we show that Snf6 and Snf5, two subunits of the SWI/SNF chromatin-remodeling complex in Saccharomyces cerevisiae, copurify with the NER damage-recognition heterodimer Rad4-Rad23. This interaction between SWI/SNF and Rad4-Rad23 is stimulated by UV irradiation. We demonstrate that NER in the transcriptionally silent, nucleosome-loaded HML locus is reduced in yeast cells lacking functional SWI/SNF. In addition, using a restriction enzyme accessibility assay, we observed UV-induced nucleosome rearrangement at the silent HML locus. Notably, this rearrangement is markedly attenuated when SWI/SNF is inactivated. These results indicate that the SWI/SNF chromatin-remodeling complex is recruited to DNA lesions by damage-recognition proteins to increase DNA accessibility for NER in chromatin.