Searching for an efficient institutional review board review model: Interrelationship of trainee-investigators, funding, and initial approval.
In this study we sought to identify variables associated with institutional review board (IRB) decisions to develop an efficient "pre-IRB" review model. We explored several variables, including relationships among the identification of trainees as investigators, external sources of funding, and initial approval. The sample consisted of all new submissions reviewed by the 2 medical IRBs at the University of Miami (UM) during a 1-year period. Trainees included students, residents, and fellows. At least 1 trainee had to be identified for a proposal to be considered a trainee submission. The medical-science committees (MSCs) were similar with regard to the numbers of new submissions they reviewed during convened meetings (MSC-A 242, MSC-B 241) and the percentages of proposals that were initially approved (MSC-A 52.9%, MSC-B 53.1%). Approved submissions were defined as those initially approved or conditionally approved pending minor modifications. We noted a robust statistical difference between the percentages of trainee submissions initially approved (39.9%) and submissions that did not identify a trainee (59.4%) ( P <.0001). Of the proposals that were initially not approved (tabled [deferred] or rejected [not approved]), 28.9% of those including a trainee were rejected, compared with 11.0% without a trainee ( P <.001). Proposals in which the source of funding was identified were more likely to be approved (64.2%) than were those in which it was not (30.8%) ( P <.0001). Funding also seemed to influence the trainee and initial-approval interaction. Our results show that new submissions that identified trainee investigators were more likely to be deferred or not approved than those that did not. Nonapproved proposals that identified a trainee were 3.8 times more likely to be initially rejected than those that did not. A prereview model could target those submissions that list a trainee, lack funding, or both.


Infrared and Raman screening of seized novel psychoactive substances: a large scale study of >200 samples.
The potential of IR absorption and Raman spectroscopy for rapid identification of novel psychoactive substances (NPS) has been tested using a set of 221 unsorted seized samples suspected of containing NPS. Both IR and Raman spectra showed large variation between the different sub-classifications of NPS and smaller, but still distinguishable, differences between closely related compounds within the same class. In initial tests, screening the samples using spectral searching against a limited reference library allowed only 41% of the samples to be fully identified. The limiting factor in the identification was the large number of active compounds in the seized samples for which no reference vibrational data were available in the libraries rather than poor spectral quality. Therefore, when 33 of these compounds were independently identified by NMR and mass spectrometry and their spectra used to extend the libraries, the percentage of samples identified by IR and Raman screening alone increased to 76%, with only 7% of samples having no identifiable constituents. This study, which is the largest of its type ever carried out, therefore demonstrates that this approach of detecting non-matching samples and then identifying them using standard analytical methods has considerable potential in NPS screening since it allows rapid identification of the constituents of the majority of street quality samples. Only one complete feedback cycle was carried out in this study but there is clearly the potential to carry out continuous identification/updating when this system is used in operational settings.


High levels of ephrinB2 over-expression increases the osteogenic differentiation of human mesenchymal stem cells and promotes enhanced cell mediated mineralisation in a polyethyleneimine-ephrinB2 gene-activated matrix.
Gene therapy can be combined with tissue engineering constructs to produce gene-activated matrices (GAMs) with enhanced capacity for repair. Polyethyleneimine (PEI), a non-viral vector, has previously been optimised for high efficiency gene transfer in rat mesenchymal stem cells (rMSCs). The use of PEI to transfect human MSCs (hMSCs) with ephrinB2 is assessed here. Recently a role for the ephrinB2 ligand and EphB4 receptor duo has been proposed in bone remodelling. Herein, over-expression of the ephrinB2 ligand resulted in increased osteogenic differentiation in hMSCs. As ephrinB2 is a cell surface anchored ligand which only interacts with cells expressing the cognate EphB4 receptor through direct contact, we have shown that direct cell-cell contact between two neighbouring cells is responsible for enhanced osteogenesis. In an effort to begin to elucidate the molecular mechanisms at play downstream of ephrinB2 over-expression, RT-PCR was performed on the GAMs which revealed no significant changes in runx2 or BMP2 expression but an upregulation of osterix (Osx) and Dlx5 expression prompting the belief that the mode of osteogenesis is independent of the BMP2 pathway. This select interaction, coupled with the transient gene expression profile of PEI, makes the PEI-ephrinB2 GAM an ideal candidate matrix for a bone targeted GAM.


A Pilot Study on the Influence of Facial Expression on Measurements in Three-Dimensional Digital Surfaces of the Face in Infants With Cleft Lip and Palate.
Three-dimensional surface imaging is an increasingly popular modality for face measurements in infants with cleft lip and palate. Infants are noncompliant toward producing specific facial expressions, and selecting the appropriate moment of acquisition is challenging. The objective was to estimate amount and spatial distribution of deformation of the face due to facial expression in infants with cleft lip and palate and provide recommendations for an improved acquisition protocol, including a method of quality control in terms of obtaining images with true neutral expression. Three-dimensional surface images of ten 4-month-old infants with unrepaired cleft lip and palate were obtained using a 3dMDface stereophotogrammetric system. For each subject, five surface images judged as representing a neutral expression were obtained during the same photo session. Mean and maximum deformations were calculated. A formalized review was performed, allowing the image exhibiting the "best" neutral expression to be selected, thus decreasing errors due to residual facial expression. Deformation due to facial expression generally increased from forehead to chin. The amount of deformation in three selected regions were determined: nose (mean, 1 mm; maximum = 3 mm); cleft region (mean, 2 mm; maximum = 5 mm); chin region (mean, 5 mm; maximum = 12 mm). Analysis indicated that introduction of a formalized review of images could reduce these errors by a factor of 2. The continuous change of facial expression in infants represents a substantial source of error; however, this may be reduced by incorporating a formalized review into the acquisition protocol.


Skin antisepsis with chlorhexidine-alcohol versus povidone iodine-alcohol, with and without skin scrubbing, for prevention of intravascular-catheter-related infection (CLEAN): an open-label, multicentre, randomised, controlled, two-by-two factorial trial.
Intravascular-catheter-related infections are frequent life-threatening events in health care, but incidence can be decreased by improvements in the quality of care. Optimisation of skin antisepsis is essential to prevent short-term catheter-related infections. We hypothesised that chlorhexidine-alcohol would be more effective than povidone iodine-alcohol as a skin antiseptic to prevent intravascular-catheter-related infections. In this open-label, randomised controlled trial with a two-by-two factorial design, we enrolled consecutive adults (age ≥18 years) admitted to one of 11 French intensive-care units and requiring at least one of central-venous, haemodialysis, or arterial catheters. Before catheter insertion, we randomly assigned (1:1:1:1) patients via a secure web-based random-number generator (permuted blocks of eight, stratified by centre) to have all intravascular catheters prepared with 2% chlorhexidine-70% isopropyl alcohol (chlorhexidine-alcohol) or 5% povidone iodine-69% ethanol (povidone iodine-alcohol), with or without scrubbing of the skin with detergent before antiseptic application. Physicians and nurses were not masked to group assignment but microbiologists and outcome assessors were. The primary outcome was the incidence of catheter-related infections with chlorhexidine-alcohol versus povidone iodine-alcohol in the intention-to-treat population. This study is registered with ClinicalTrials.gov, number NCT01629550 and is closed to new participants. Between Oct 26, 2012, and Feb 12, 2014, 2546 patients were eligible to participate in the study. We randomly assigned 1181 patients (2547 catheters) to chlorhexidine-alcohol (594 patients with scrubbing, 587 without) and 1168 (2612 catheters) to povidone iodine-alcohol (580 patients with scrubbing, 588 without). Chlorhexidine-alcohol was associated with lower incidence of catheter-related infections (0·28 vs 1·77 per 1000 catheter-days with povidone iodine-alcohol; hazard ratio 0·15, 95% CI 0·05-0·41; p=0·0002). Scrubbing was not associated with a significant difference in catheter colonisation (p=0·3877). No systemic adverse events were reported, but severe skin reactions occurred more frequently in those assigned to chlorhexidine-alcohol (27 [3%] patients vs seven [1%] with povidone iodine-alcohol; p=0·0017) and led to chlorhexidine discontinuation in two patients. For skin antisepsis, chlorhexidine-alcohol provides greater protection against short-term catheter-related infections than does povidone iodine-alcohol and should be included in all bundles for prevention of intravascular catheter-related infections. University Hospital of Poitiers, CareFusion.


Supporting implementation of Cochrane methods in complex communication reviews: resources developed and lessons learned for editorial practice and policy.
Every healthcare encounter involves some form of communication and there is growing recognition that effective health communication is central to the delivery of safe, high-quality healthcare. Conversely, poor communication has a range of adverse consequences for those receiving healthcare and the systems delivering care, including elevated patient safety risks. Increasing understanding and documentation of the key role that good communication plays in healthcare design and delivery has meant there is growing demand from policy-makers and other decision-makers for evidence on the effects of health communication interventions - that is, how best to communicate. While systematic reviews of such interventions are fundamental to building this evidence base, such interventions and reviews are often highly complex and pose considerable challenges for authors and editors. In this paper, we describe our experience as a Cochrane editorial group identifying common issues in reviews of communication interventions and developing resources to support authors to better meet these challenges. Our analysis found that issues typically fell into one or more of the following three stages of the review process: understanding and applying systematic review methods (e.g. selecting outcomes for analysis); reporting the review's methods (e.g. describing key decisions made in conducting the review); and interpreting the findings (e.g. incorporating quality of the evidence into findings of the review). We also found that common issues reflected both practical difficulties (such as the typically large size of reviews and disparate measures for outcomes) and conceptual challenges (for instance, the difficulties of identifying comparisons). While extensive advice for Cochrane systematic reviewers exists, this standardised advice does not cover all of the issues emerging for complex communication reviews. In response, we therefore developed a collection of resources, both general and targeted to specific methodological issues. Here, we describe the types of resources developed and the aims of these, the rationale for why we needed to fill specific gaps in existing advice, and reflect on the lessons for future editorial practice, policies and research in relation to the implementation of Cochrane review methods in the area of health communication.


Skeletal morbidity in children receiving chemotherapy for acute lymphoblastic leukemia and its association with mineral homeostasis and duration of inpatient stay.
Reduced activity, older age, and abnormal bone mineral status are considered as important determinants of poor bone health in children with acute lymphoblastic leukemia (ALL). The independent contribution of these factors toward skeletal morbidity (SM) requires further investigation. The aim of this study was to investigate the influence of activity, age, and mineral status over the first 12 months of chemotherapy on subsequent SM. The medical records of 56 children presenting with ALL between 2003 and 2007 and treated on UKALL2003 were reviewed for the number of inpatient days over the first 12 months of chemotherapy as a surrogate marker of inactivity and lack of well-being. Data for serum Ca, albumin, Mg, and Pho were also collected over this period. SM was defined as any episode of musculoskeletal pain or fractures. The median duration of inpatient days over the first 12 months of treatment in children with no SM was 58 days (40,100), whereas the median number of inpatient days during the first 12 months in those children with any SM, musculoskeletal pain only, or fractures only was 83 days (54,131), 81 days (52,119) and 91 days (59,158), respectively (P=0.003). Children with SM and fractures particularly had lower levels of serum Ca, Mg, and Pho compared with those without SM over the first 12 months of chemotherapy. There was a higher risk of SM in those who were diagnosed after the age of 8 years (P=0.001, odds ratio=16, 95% confidence interval: 3.80). Multiple regression analysis showed that the incidence of SM only had a significant independent association with age at diagnosis (P=0.001) and the number of inpatient days (P=0.03) over the first 12 months (r=23). All children who were diagnosed after the age of 8 years with an inpatient stay of more than 75 days, in the first 12 months of the chemotherapy (n,14) children had some form of SM (odds ratio=64). The incidence of SM in children receiving chemotherapy for ALL is associated with a higher likelihood of being older and having longer periods of inpatient stay. The close link between age and changes in bone mineral status may be one explanation for the increased bone morbidity in ALL children.


Medium chain triglycerides and vitamin E reduce the severity of established experimental alcoholic liver disease.
Lipid peroxidation may be important in the pathogenesis of alcoholic liver injury. We investigated the potential of medium chain triglycerides and vitamin E to decrease lipid peroxidation and reverse established alcoholic liver injury. Four groups (five rats/group) of male Wistar rats were studied. Rats in group 1 were fed a fish oil-ethanol diet for 6 weeks. Rats in groups 2, 3 and 4 were fed the fish oil-ethanol diet for 6 weeks before being switched to fish oil-dextrose (group 2), fish oil-dextrose plus vitamin E (group 3) or medium chain triglycerides-dextrose (group 4) diets for 2 weeks. Liver samples were analyzed for histopathology, lipid peroxidation, fatty acid composition and cytochrome P450 2E1 activity. By 6 weeks, all rats developed fatty liver, inflammation and necrosis. After switching to the dextrose-containing diets, there was minimal histologic improvement in group 2, moderate improvement in group 3 and near normalization of the histology in group 4. Histologic improvement was associated with decreased lipid peroxidation and cytochrome P450 2E1 activity. Higher levels of polyunsaturated fatty acids were seen in groups 2 and 3 than in group 4. Our results indicate that a diet enriched in saturated (group 4) but not polyunsaturated (group 2) fatty acids effectively reverses alcoholic liver injury. Treatment with vitamin E also led to histologic improvement. These effects may be explained, at least in part, by down-regulation of lipid peroxidation. Other effects of medium chain triglycerides, such as their propensity for oxidation rather than esterification, may also be important.


[The assessment of the clinical efficacy, vasoactive and metabolic effects of mexidol in elderly patients with discirculatory encephalopathy].
Forty patients, aged from 55 to 74 years, with chronic cerebral ischemia - discirculatory encephalopathy, stages I--II, against the background of arterial hypertension and atherosclerosis were studied. Patients were randomized into 2 equal groups. Patients of the main group received the antioxidant mexidol intramuscular in dose 100 mg during the first 10 days and then perorally in dose 125 mg three times a day during 20 days in the combination with the regular treatment (diroton in dose 5-20 mg daily, cardiomagnyl in dose 75 mg daily). Patients of the control group received regular treatment during 30 days. The efficacy was evaluated at baseline, on the 10th and 30th days after the treatment using clinical scales (the number of complaints, neurological deficit, cognitive and emotional disorders), parameters of cerebral hemodynamics measured with transcranial duplex scanning, laboratory indices of the state of oxidant and antioxidant systems (the content of malonic dialdehyde and superoxide dismutase in the blood plasma as well as the plasma total antioxidant activity). The statistically significant (p<0.05-0.01) reduction in the severity of asthenic and anxiety disorders, the improvement of static-locomotor and cognitive functions as well as of parameters of cerebral hemodynamics due to the decrease in the peripheral vascular resistance and the normalization of venous outflow from the cavity of scull was seen in patients treated with mexidol compared to the baseline and the control group. Mexidol significantly reduced the intensity of lipid peroxidation and increase the total antioxidant activity of the blood. The results obtained in the study allow to consider mexidol as the effective drug in the complex treatment of discirculatory encephalopathy in elderly patients.


Viral and molecular aspects of oral cancer.
Squamous cell carcinoma (SCC) is the most common epithelial malignancy in the oral cavity. SCCs and their variants constitute over 90% of oral malignancies, and the disease is associated with poor prognosis. OSCC is a complex malignancy where environmental factors, virus infections, and genetic alterations most likely interact, and thus give rise to the malignant condition. Herein, we review the available literature regarding high-risk factors such as alcohol and tobacco usage; discuss the roles of human papillomaviruses (HPV), the Epstein-Barr virus, and the human herpes simplex virus (HSV); and evaluate several candidate genes associated with the condition: p53, p16(INK4) and p21(WAF1/CIPI), survivin, B-cell lymphoma-2 (BCL-2), keratins, Fibroblast growth factor 3 (FGF3), FGF4, FGF19, Oral cancer overexpressed gene 1 (ORAOV1), and Cyclin D1 (CCND1).


Long-term patterns of shrub expansion in a C4-dominated grassland: fire frequency and the dynamics of shrub cover and abundance.
Worldwide, grassland ecosystems have experienced a major shift in growth-form dominance as woody plant species have expanded and replaced native grasses. In the C(4)-dominated grasslands of central North America, a reduction in fire frequency is the most cited cause of this shift in growth forms as fire both enhances grass productivity and constrains the establishment and expansion of native woody vegetation. Using an 18-yr plant species composition data set, we quantified patterns of change in shrub cover, frequency, and species richness associated with three distinct fire regimes. During the study period (1983-2000), shrub cover increased most dramatically in sites in which the frequency of fire was once every 4 yr (intermediate frequency; 28.6%) followed by sites in which fire occurred only once during the 18-yr period (low frequency; 23.7%). Annual fire effectively prevented the recruitment of new woody species, but even with this high fire frequency, shrub cover increased slightly (3.7%). Comparatively, shrub species richness increased by three and six, respectively, in the intermediate- and low-frequency fire sites. These data indicate that within this grassland, periods without fire are necessary for recruitment of both new individuals and additional shrub species; however, once established, shrub cover will increase regardless of fire frequency and even annual fire will not reduce shrub abundance.


High variability in strain estimation errors when using a commercial ultrasound speckle tracking algorithm on tendon tissue.
Ultrasound speckle tracking offers a non-invasive way of studying strain in the free Achilles tendon where no anatomical landmarks are available for tracking. This provides new possibilities for studying injury mechanisms during sport activity and the effects of shoes, orthotic devices, and rehabilitation protocols on tendon biomechanics. To investigate the feasibility of using a commercial ultrasound speckle tracking algorithm for assessing strain in tendon tissue. A polyvinyl alcohol (PVA) phantom, three porcine tendons, and a human Achilles tendon were mounted in a materials testing machine and loaded to 4% peak strain. Ultrasound long-axis cine-loops of the samples were recorded. Speckle tracking analysis of axial strain was performed using a commercial speckle tracking software. Estimated strain was then compared to reference strain known from the materials testing machine. Two frame rates and two region of interest (ROI) sizes were evaluated. Best agreement between estimated strain and reference strain was found in the PVA phantom (absolute error in peak strain: 0.21 ± 0.08%). The absolute error in peak strain varied between 0.72 ± 0.65% and 10.64 ± 3.40% in the different tendon samples. Strain determined with a frame rate of 39.4 Hz had lower errors than 78.6 Hz as was the case with a 22 mm compared to an 11 mm ROI. Errors in peak strain estimation showed high variability between tendon samples and were large in relation to strain levels previously described in the Achilles tendon.


Fournier's gangrene: outcome analysis and prognostic factors.
Fournier's gangrene is an infective necrotising fasciitis of the external genitalia and perineum associated with significant morbidity and mortality. The factors associated with non survival have been described but are not universally accepted. The identification of prognostic factors remains critical to improve outcomes. To determine the hospital based mortality and factors associated with non-survival among subjects with a clinical diagnosis of Fournier's gangrene. A prospective hospital based observational study on 51patients with a clinical diagnosis of Fournier's gangrene over a 2-year period. A comparison was made between survivors and non-survivors to establish prognostic factors associated with non survival. The disease related hospital mortality was 27% (14/51). The mean age of the 51, all male patients was 47 years. An older age was significantly associated with non-survival (p=0.02). The presence of renal dysfunction (p=0.001), severe sepsis (p=0.000), delay in surgical debridment (p=0.04), urogenital source of infection (p=0.01), a body surface area involvement of greater than 5% (p=0.006), abdominal wall involvement (p=0.02) on admission were significant factors associated with mortality. The presence of either HIV infection or diabetes mellitus was not a prognostic indicator of mortality. The clinical and biochemical parameters on admission associated with non survival were a high respiratory rate (p=0.03), a low hemoglobin(p=0.0001), an elevated blood urea nitrogen (p=0.005) and creatinine (p=0.01). Multivariate logistic regression analysis did not show any independent factors associated with non survival. Fournier's gangrene remains a fatal condition with a hospital mortality of 27%. Prognostic factors for non survival include an advanced age, a urogenital source of infection, abdominal involvement, severe sepsis and renal dysfunction.


The Role of medicinal herbs in treatment of insulin resistance in patients with Polycystic Ovary Syndrome: A literature review.
Background Polycystic Ovary Syndrome (PCOS) is one of the most common endocrine abnormalities in women. Due to the side effects of drugs, the tendency to use natural antioxidants and anti-inflammatory agents to regulate metabolism, hyperinsulinemia, and hyperlipidemia in PCOS patients has been increased. This review aimed to investigate the role of herbal substances on the treatment of PCOS. Methods The present review was carried out using keywords such as polycystic ovary syndrome and/or PCOS and/or herb. Databases including Web of Science, PubMed, and Science Direct were used to collect all related articles published from 1990 to 2019. We excluded studies unrelated to the PCOS and medical herbs. Results Overall, 361 records were identified through database searching. After primary screening and the full-texts assessment, 323 records were excluded, and 38 articles were finally included. The results indicate that some medicinal herbs may have a key role in treating PCOS. The compounds in these medical herbs can affect lipid profiles (Aloe vera, chamomile, and cinnamon), insulin resistance (cinnamon, chamomile, Aloe vera, and Camellia sinensis), blood glucose (Aloe vera, cinnamon, and Camellia sinensis), hormones (Aloe vera, silymarin, chamomile, fenugreek, Camellia sinensis, Heracleum persicum, Potentilla, Mentha spicata, Foeniculum vulgar, licorice, and Marrubium), and ovarian tissue (Aloe vera, chamomile, Camellia sinensis, Mentha spicata, and silymarin). Conclusion Natural substances such as Aloe vera, cinnamon, green tea, fenugreek, and silymarin can be used as a new supportive care for PCOS. Further clinical trials are warranted to confirm their benefits and safety.


Increased expression of phospholipase D in the heart with experimental autoimmune myocarditis in Lewis rats.
The expression of phospholipase D (PLD) in the hearts of rats with experimental autoimmune myocarditis (EAM) was studied to elucidate the functional role of PLD in the pathogenesis of EAM. Western blot analysis showed that the level of the PLD1 isoform was significantly increased in the hearts of rats with EAM on days 14, 17 and 21 postimmunization (pi) (P < 0.01; control vs EAM at 14 pi, 17 pi and 21 pi). The phenotypes of cells exhibiting increased PLD1 expression were primarily inflammatory cells, including ED1 positive macrophages, in the inflammatory EAM lesions. Some cardiomyocytes also showed increased PLD1 immunoreaction in and around EAM lesions. Some PLD1-positive cells were also positive for proliferating cell nuclear antigen in some cardiomyocytes or terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling in some macrophages, suggesting that PLD1 positive cells have a capacity for proliferation or apoptosis depending on cell types in the target organ. Thus, it is postulated that increased expression of PLD1 in EAM may support an early inflammatory response in proliferating inflammatory cells, and its expression in cardiomyocytes may help them to survive by activation of survival factors in hearts with EAM.


Different genes interact with particulate matter and tobacco smoke exposure in affecting lung function decline in the general population.
Oxidative stress related genes modify the effects of ambient air pollution or tobacco smoking on lung function decline. The impact of interactions might be substantial, but previous studies mostly focused on main effects of single genes. We studied the interaction of both exposures with a broad set of oxidative-stress related candidate genes and pathways on lung function decline and contrasted interactions between exposures. For 12679 single nucleotide polymorphisms (SNPs), change in forced expiratory volume in one second (FEV(1)), FEV(1) over forced vital capacity (FEV(1)/FVC), and mean forced expiratory flow between 25 and 75% of the FVC (FEF(25-75)) was regressed on interval exposure to particulate matter <10 µm in diameter (PM10) or packyears smoked (a), additive SNP effects (b), and interaction terms between (a) and (b) in 669 adults with GWAS data. Interaction p-values for 152 genes and 14 pathways were calculated by the adaptive rank truncation product (ARTP) method, and compared between exposures. Interaction effect sizes were contrasted for the strongest SNPs of nominally significant genes (p(interaction)<0.05). Replication was attempted for SNPs with MAF>10% in 3320 SAPALDIA participants without GWAS. On the SNP-level, rs2035268 in gene SNCA accelerated FEV(1)/FVC decline by 3.8% (p(interaction) = 2.5×10(-6)), and rs12190800 in PARK2 attenuated FEV1 decline by 95.1 ml p(interaction) = 9.7×10(-8)) over 11 years, while interacting with PM10. Genes and pathways nominally interacting with PM10 and packyears exposure differed substantially. Gene CRISP2 presented a significant interaction with PM10 (p(interaction) = 3.0×10(-4)) on FEV(1)/FVC decline. Pathway interactions were weak. Replications for the strongest SNPs in PARK2 and CRISP2 were not successful. Consistent with a stratified response to increasing oxidative stress, different genes and pathways potentially mediate PM10 and tobacco smoke effects on lung function decline. Ignoring environmental exposures would miss these patterns, but achieving sufficient sample size and comparability across study samples is challenging.


Over-the-counter analgesics: a toxicology perspective.
The decision to use any analgesic is a balance of benefit and risk. In the case of analgesics, it is important to balance the therapeutic benefit against both the risk in therapeutic use and the risk (and ease of treatment) in overdose. Paracetamol in therapeutic dose carries little risk of adverse events. Less than 0.1% of the estimated 30 million paracetamol users in the United Kingdom attend hospital with a paracetamol overdose each year, and approximately 200 people die, most of whom presented late or did not receive antidote, N-acetylcysteine, within 12 hours. Nonsteriodal anti-inflammatory drugs (NSAIDs) have greater adverse effects in therapeutic use than paracetamol but also have a lower incidence of severe features or death in overdose. There is no antidote available for NSAID poisoning. Aspirin carries both significant adverse effects in therapeutic dose and a substantial risk in overdose, for which there is no antidote. Its risk-benefit profile is probably the poorest of all analgesics currently available over-the-counter (OTC); this is reflected in current trends both in analgesic use and overdose figures. Although a number of options to reduce deaths from poisoning by OTC analgesics have been considered, few are practical, and all must take account of the public health benefits provided by these drugs. A perspective should be retained that the vast majority of the population in Australia, the United States, the United Kingdom, and Denmark derive therapeutic benefit from OTC analgesics and do not take them in overdose. The majority of those who do take overdoses come to little or no harm. Management of serious poisoning by paracetamol, aspirin, or NSAIDs remains a medical challenge.


Effects of Wharton's jelly-derived mesenchymal stem cells on neonatal neutrophils.
Mesenchymal stem cells (MSCs) have been proposed as autologous therapy for inflammatory diseases in neonates. MSCs from umbilical cord Wharton's jelly (WJ-MSCs) are accessible, with high proliferative capacity. The effects of WJ-MSCs on neutrophil activity in neonates are not known. We compared the effects of WJ-MSCs on apoptosis and the expression of inflammatory, oxidant, and antioxidant mediators in adult and neonatal neutrophils. WJ-MSCs were isolated, and their purity and function were confirmed by flow cytometry. Neutrophils were isolated from cord and adult blood by density centrifugation. The effects of neutrophil/WJ-MSC co-culture on apoptosis and gene and protein expression were measured. WJ-MSCs suppressed neutrophil apoptosis in a dose-dependent manner. WJ-MSCs decreased gene expression of NADPH oxidase-1 in both adult and neonatal neutrophils, but decreased heme oxygenase-1 and vascular endothelial growth factor and increased catalase and cyclooxygenase-2 in the presence of lipopolysaccharide only in adult cells. Similarly, generation of interleukin-8 was suppressed in adult but not neonatal neutrophils. Thus, WJ-MSCs dampened oxidative, vascular, and inflammatory activity by adult neutrophils, but neonatal neutrophils were less responsive. Conversely, Toll-like receptor-4, and cyclooxygenase-2 were upregulated in WJ-MSCs only in the presence of adult neutrophils, suggesting an inflammatory MSC phenotype that is not induced by neonatal neutrophils. Whereas WJ-MSCs altered gene expression in adult neutrophils in ways suggesting anti-inflammatory and antioxidant effects, these responses were attenuated in neonatal cells. In contrast, inflammatory gene expression in WJ-MSCs was increased in the presence of adult but not neonatal neutrophils. These effects should be considered in clinical trial design before WJ-MSC-based therapy is used in infants.


Dextran sulphate-induced tau assemblies cause endogenous tau aggregation and propagation in wild-type mice.
Accumulation of assembled tau protein in the central nervous system is characteristic of Alzheimer's disease and several other neurodegenerative diseases, called tauopathies. Recent studies have revealed that propagation of assembled tau is key to understanding the pathological mechanisms of these diseases. Mouse models of tau propagation are established by injecting human-derived tau seeds intracerebrally; nevertheless, these have a limitation in terms of regulation of availability. To date, no study has shown that synthetic assembled tau induce tau propagation in non-transgenic mice. Here we confirm that dextran sulphate, a sulphated glycosaminoglycan, induces the assembly of recombinant tau protein into filaments in vitro. As compared to tau filaments induced by heparin, those induced by dextran sulphate showed higher thioflavin T fluorescence and lower resistance to guanidine hydrochloride, which suggests that the two types of filaments have distinct conformational features. Unlike other synthetic filament seeds, intracerebral injection of dextran sulphate-induced assemblies of recombinant tau caused aggregation of endogenous murine tau in wild-type mice. AT8-positive tau was present at the injection site 1 month after injection, from where it spread to anatomically connected regions. Induced tau assemblies were also stained by anti-tau antibodies AT100, AT180, 12E8, PHF1, anti-pS396 and anti-pS422. They were thioflavin- and Gallyas-Braak silver-positive, indicative of amyloid. In biochemical analyses, accumulated sarkosyl-insoluble and hyperphosphorylated tau was observed in the injected mice. In conclusion, we revealed that intracerebral injection of synthetic full-length wild-type tau seeds prepared in the presence of dextran sulphate caused tau propagation in non-transgenic mice. These findings establish that propagation of tau assemblies does not require tau to be either mutant and/or overexpressed.


A refined model for the somatostatin pharmacophore: conformational analysis of lanthionine-sandostatin analogs.
We report the conformational analysis of a series of analogs of sandostatin (octreotide, D-Phe1-c[Cys2-Phe3-D-Trp4-Lys5-Thr6-Cys 7]-Thr8-ol) using 1H NMR spectroscopy and molecular modeling. Two active compounds in which the disulfide group is replaced by a monosulfide (lanthionine) bridge (D-Phe1-c[AlaL2-Phe3-D-Trp4-Lys5-Thr6-A laL7]-Thr8-ol and D-Phe1-c[AlaL2-Phe3-D-Trp4-Lys5-Thr6-Al aL7]-Thr8-NH2, where AlaL denotes each of the lanthionine amino acid ends linked by the monosulfide bridge) show different mSSTR2b/rSSTR5 receptor selectivities as compared to sandostatin. These new results have enabled us to reveal features of the somatostatin pharmacophore common to the model previously proposed in our laboratory on the basis of main chain and side chain chiral methylation studies. In addition, our studies provide new insight into the role of the disulfide bridge and of Thr8 in binding potency. We also show that the lanthionine group is a good mimetic of beta-VI turns and can be incorporated in sandostatin analogs maintaining the essential secondary structural features of sandostatin. These results facilitate the design of new sandostatin peptidomimetics.


Antisense DNA and RNA agents against picornaviruses.
Anti-picornaviral antisense agents are part of a broader group of nucleic acid-based molecules developed for sequence-specific inhibition of translation and/or transcription of the target sequence through induced nuclease activity or physical hindrance. Three types of nucleic acid-based gene silencing molecules can be distinguished, including DNA-base antisense oligonucleotides (ASO), nucleic acid enzymes (ribozyme and DNAzyme) and double-stranded small interfering RNA (siRNA or microRNA). These antisense DNA and RNA molecules have been widely studied for gene functional studies and therapeutic purposes. In this review, we focus on drug development using ASO and siRNA strategies to inhibit picornavirus infections. The picornavirus genome organization and life cycle is described, followed by discussion of design considerations, chemical modifications and drug delivery approaches. Recent studies using antisense against picornavirus are reviewed. Finally, we compare the advantages and disadvantages of the antisense agents with those of other therapeutics, taking into consideration their limitations which need to be overcome to achieve the final goal of clinical application.


Comparison of compounds of three Rubus species and their antioxidant activity.
Rubus amabilis, Rubus niveus Thunb., and Rubus sachalinensis are three Rubus species that are alternatively found in Manubzhithang, a Tibetan medicine, in different areas of China. The current study analyzed HPLC/UV chromatograms and it compared compounds of these three Rubus species in contrast to reference substances such as 2,6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside, procyanidin B4, and isovitexin-7-O-glucoside. The three Rubus species produced similar peaks in chromatograms. The antioxidant activity of the three Rubus species was determined using an assay for DPPH free radical scavenging activity. Results indicated that the three Rubus species extracts had almost the same level of free radical scavenging activity. Thus, findings indicated the rationality of substituting these species for one another as an ingredient in Manubzhithang.


Diffusion-weighted multiple shot echo planar imaging of humans without navigation.
A new method is proposed for reducing the artifacts produced in diffusion-weighted imaging. When data are acquired using multiple shot echo planar acquisitions, conventional reconstruction methods produce artifactual images as a consequence of diffusion weighting and small amounts of bulk motion of the subject. If the amount of motion can be determined, it is possible to correct the data before reconstruction, which removes the artifact. A method for estimating the motion from the acquired data has been developed and evaluated. This method assumes that ghost image effects will be minimized when motion has been correctly compensated. By considering the amount of signal in the background of the image, appropriate corrections to the data can be made, and the accuracy of the motion compensation may be estimated. This technique has been evaluated by computer simulation, and its performance has been demonstrated in a phantom and humans with both two- and four-shot echo planar acquisitions and using both "mosaic" and "interleaved" sampling schemes.


[Determination of a visco-hyperelastic material law based on dynamic tension test data].
The objective of this study was to determine the visco-hyperelastic constitutive law of brain tissue under dynamic impacts. A method combined by finite element simulations and optimization algorithm was employed for the determination of material variables. Firstly, finite element simulations of brain tissue dynamic uniaxial tension, with a maximum stretch rate of 1.3 and strain rates of 30 s -1 and 90 s -1, were developed referring to experimental data. Then, fitting errors between the engineering stress-strain curves predicted by simulations and experimental average curves were assigned as objective functions, and the multi-objective genetic algorithm was employed for the optimation solution. The results demonstrate that the brain tissue finite element models assigned with the novel obtained visco-hyperelastic material law could predict the brain tissue's dynamic mechanical characteristic well at different loading rates. Meanwhile, the novel material law could also be applied in the human head finite element models for the improvement of the biofidelity under dynamic impact loadings.


Gain-of-function of mutated C-CBL tumour suppressor in myeloid neoplasms.
Acquired uniparental disomy (aUPD) is a common feature of cancer genomes, leading to loss of heterozygosity. aUPD is associated not only with loss-of-function mutations of tumour suppressor genes, but also with gain-of-function mutations of proto-oncogenes. Here we show unique gain-of-function mutations of the C-CBL (also known as CBL) tumour suppressor that are tightly associated with aUPD of the 11q arm in myeloid neoplasms showing myeloproliferative features. The C-CBL proto-oncogene, a cellular homologue of v-Cbl, encodes an E3 ubiquitin ligase and negatively regulates signal transduction of tyrosine kinases. Homozygous C-CBL mutations were found in most 11q-aUPD-positive myeloid malignancies. Although the C-CBL mutations were oncogenic in NIH3T3 cells, c-Cbl was shown to functionally and genetically act as a tumour suppressor. C-CBL mutants did not have E3 ubiquitin ligase activity, but inhibited that of wild-type C-CBL and CBL-B (also known as CBLB), leading to prolonged activation of tyrosine kinases after cytokine stimulation. c-Cbl(-/-) haematopoietic stem/progenitor cells (HSPCs) showed enhanced sensitivity to a variety of cytokines compared to c-Cbl(+/+) HSPCs, and transduction of C-CBL mutants into c-Cbl(-/-) HSPCs further augmented their sensitivities to a broader spectrum of cytokines, including stem-cell factor (SCF, also known as KITLG), thrombopoietin (TPO, also known as THPO), IL3 and FLT3 ligand (FLT3LG), indicating the presence of a gain-of-function that could not be attributed to a simple loss-of-function. The gain-of-function effects of C-CBL mutants on cytokine sensitivity of HSPCs largely disappeared in a c-Cbl(+/+) background or by co-transduction of wild-type C-CBL, which suggests the pathogenic importance of loss of wild-type C-CBL alleles found in most cases of C-CBL-mutated myeloid neoplasms. Our findings provide a new insight into a role of gain-of-function mutations of a tumour suppressor associated with aUPD in the pathogenesis of some myeloid cancer subsets.


Early psychological interventions for adult survivors of trauma: a review.
Psychological interventions after traumatic events have only recently been evaluated in randomized, controlled trials. Recent systematic reviews concluded that single sessions of individual psychological debriefing are not effective in reducing distress or subsequent posttraumatic stress disorder (PTSD) symptoms. The present article reviews trials of early cognitive behavior therapy (CBT) after trauma. Cognitive behavioral therapy was more effective than supportive counseling in preventing chronicity of PTSD symptoms; however, in most available studies it remained unclear whether supportive counseling facilitated or retarded recovery, compared with no intervention. A brief CBT program given in the first month of trauma was not superior to repeated assessment; however, a course of CBT of up to 16 sessions given at 1-4 months after trauma was superior to self-help, repeated assessment, and no intervention. Possible reasons for the difference in efficacy between CBT and debriefing or self-help are discussed. These include the way of working through traumatic memories and the impact of the interventions on patients' interpretations of their PTSD symptoms. Possible ways of identifying people who are in need of specialist psychological intervention after trauma and who are unlikely to recover on their own are discussed. Some ideas for alternative ways of offering help to trauma survivors are presented, and methodologic suggestions for future research are given.


Readiness factors for lean implementation in healthcare settings--a literature review.
The purpose of this paper is to determine the readiness factors that are critical to the application and success of lean operating principles in healthcare organizations through a review of relevant literature. A comprehensive review of literature focussing on lean and lean healthcare was conducted. Leadership, organizational culture, communication, training, measurement, and reward systems are all commonly attributed readiness factors throughout general change management and lean literature. However, directly related to the successful implementation of lean in healthcare is that a setting is able to authorize a decentralized management style and undertake an end-to-end process view. These can be particularly difficult initiatives for complex organizations such as healthcare settings. The readiness factors identified are based on a review of the published literature. The external validity of the findings could be enhanced if tested using an empirical study. The readiness factors identified will enable healthcare practitioners to be better prepared as they begin their lean journeys. Sustainability of the lean initiative will be at stake if these readiness factors are not addressed. To the best of the knowledge, this is the first paper that provides a consolidated list of key lean readiness factors that can guide practice, as well as future theory and empirical research.


The ubiquitin ligase Mindbomb 1 coordinates gastrointestinal secretory cell maturation.
After cell fate specification, differentiating cells must amplify the specific subcellular features required for their specialized function. How cells regulate such subcellular scaling is a fundamental unanswered question. Here, we show that the E3 ubiquitin ligase Mindbomb 1 (MIB1) is required for the apical secretory apparatus established by gastric zymogenic cells as they differentiate from their progenitors. When Mib1 was deleted, death-associated protein kinase-1 (DAPK1) was rerouted to the cell base, microtubule-associated protein 1B (MAP1B) was dephosphorylated, and the apical vesicles that normally support mature secretory granules were dispersed. Consequently, secretory granules did not mature. The transcription factor MIST1 bound the first intron of Mib1 and regulated its expression. We further showed that loss of MIB1 and dismantling of the apical secretory apparatus was the earliest quantifiable aberration in zymogenic cells undergoing transition to a precancerous metaplastic state in mouse and human stomach. Our results reveal a mechanistic pathway by which cells can scale up a specific, specialized subcellular compartment to alter function during differentiation and scale it down during disease.


Oxidative stress-related liver dysfunction by sodium arsenite: Alleviation by Pistacia lentiscus oil.
Pistacia lentiscus L. (Anacardiaceae) is an evergreen shrub widely distributed throughout the Mediterranean region. Pistacia lentiscus oil (PLo) was particularly known in North African traditional medicine. Thus, people of these regions have used it externally to treat sore throats, burns and wounds, as well as they employed it internally for respiratory allergies. PLo is rich in essential fatty acids, vitamin E and polyphenols. As a very active site of metabolism, liver is reported to be susceptible to arsenic (As) intoxication. The present study evaluates the protective effect of PLo against sodium arsenite-induced hepatic dysfunction and oxidative stress in experimental Wistar rats. Twenty-eight rats were equally divided into four groups; the first served as a control, the remaining groups were respectively treated with PLo (3.3 mL/kg body weight), sodium arsenite (5.55 mg/kg body weight) and a combination of sodium arsenite and PLo. After 21 consecutive days, cellular functions were evaluated by hematological, biochemical and oxidative stress markers. A significant decrease in the levels of red blood cells, haemoglobin (p ≤ 0.001), hematocrit (p ≤ 0.001), reduced glutathione and metallothionein (p ≤ 0.05) associated with a significant increase of malondialdehyde (p ≤ 0.001) were noticed in the arsenic-exposed group when compared to the control. The As-treated group also exhibited an increase in hepatic antioxidant enzymes namely superoxide dismutase, glutathione peroxidase (p ≤ 0.01) and catalase (p ≤ 0.05). However, the co-administration of PLo has relatively reduced arsenic effect. The results showed that arsenic intoxication disturbed the liver pro-oxidant/antioxidant status. PLo co-administration mitigates arsenic-induced oxidative damage in rat.


Experimental demonstration of a multi-wavelength distributed feedback semiconductor laser array with an equivalent chirped grating profile based on the equivalent chirp technology.
We report, to the best of our knowledge, the first realization of a multi-wavelength distributed feedback (DFB) semiconductor laser array with an equivalent chirped grating profile based on equivalent chirp technology. All the lasers in the laser array have an identical grating period with an equivalent chirped grating structure, which are realized by nonuniform sampling of the gratings. Different wavelengths are achieved by changing the sampling functions. A multi-wavelength DFB semiconductor laser array is fabricated and the lasing performance is evaluated. The results show that the equivalent chirp technology is an effective solution for monolithic integration of a multi-wavelength laser array with potential for large volume fabrication.


Measuring vaccine acceptance among Canadian parents: A survey of the Canadian Immunization Research Network.
Parental decision making about childhood vaccinations is complex and multidimensional. There is a perception that the number of parents having concerns regarding childhood vaccinations has been increasing in Canada. The aim of this study was to explore vaccine hesitancy among Canadian parents and to examine factors associated with a parent's intention to vaccinate his/her child. Informed by the Theory of Planned Behaviour (TPB) this study assesses potential associations between parents' knowledge, attitudes and beliefs toward vaccination and their intention to vaccinate their child in the future. A national sample of Canadian parents of children aged 24-59 months (N = 2013) was surveyed using an online survey methodology. Half of the surveyed parents strongly intended to have their child vaccinated in the future. Parents' information needs and searches as well as parents' trust in different institutions were associated with intention to vaccinate. Parents who reported having frequently looked for vaccine information, who considered that it was their role as parents to question vaccines, or who had previously experienced difficulty accessing vaccination services were less likely to strongly intend to vaccinate their child in the future. Parents who had a high level of trust in doctors and public health were most likely to strongly intend to vaccinate their child. Results of the multivariate analysis showed that positive attitudes (aOR = 8.0; 95% CI: 6.0, 10.4), higher perceived social support (aOR = 3.0; 95% CI: 2.3, 3.93), and higher perceived behavioural control (aOR = 1.8; 95% CI: 1.4, 2.43) were associated with parents' intention to vaccinate their child. Findings of this study suggest that trust-building interventions that promote pro-vaccine social norms and that address negative attitudes toward vaccination could enhance vaccine acceptance among Canadian parents.


Antibody-guided radiolocalisation of tumours in patients with testicular or ovarian cancer using two radioiodinated monoclonal antibodies to placental alkaline phosphatase.
Monoclonal antibodies (H317 and H17E2) against placental-type alkaline phosphatase and testicular placental-like alkaline phosphatase were radiolabelled with 123I or 131I and used in a prospective study of patients with germ-cell neoplasms of the testis (13 studies) or epithelial-origin neoplasms of the ovary (13 studies). The presence of tumour was confirmed in the majority of patients with active disease by antibody scanning. The absence of tumour was confirmed in all cases in patients in complete remission. In two patients with positive antibody scans but no disease found by conventional radiological evaluation, further investigations subsequently revealed the presence of tumour. A positive antibody scan indicates the definite presence of a tumour, although a negative antibody scan does not always exclude the presence of disease. This method may be of clinical value in establishing disease status in patients with potentially curable diseases such as ovarian or testicular cancers.


Experimental method to characterize the strain dependent permeability of tissue engineering scaffolds.
Permeability is an overarching mechanical parameter encompassing the effects of porosity, pore size, and interconnectivity of porous structures. This parameter directly influences transport of soluble particles and indirectly regulates fluid pressure and velocity in tissue engineering scaffolds. The permeability also contributes to the viscoelastic behavior of visco-porous material under loading through frictional drag mechanism. We propose a straightforward experimental method for permeability characterization of tissue engineering scaffolds. In the developed set-up a step-wise spacer was designed to facilitate measurement of the permeability under different compressive strains while maintaining similar experimental conditions during the successive measurements. As illustration of the method, we measured the permeability of scaffolds presenting different average pore sizes and subjected to different compression values. Results showed an exponential relationship between the permeability and the average pore size of the scaffolds. Furthermore, the trend of the permeability decrease with compressive strains was depending on pore sizes of the scaffolds. The permeability also appeared to play a role in relaxation behavior of the scaffolds.


Direct carbonization of organic solvents toward graphene quantum dots.
The bottom-up synthesis of graphene quantum dots (GQDs) using solvothermal methods has attracted considerable attention because of their fewer defects and controllable size/morphology. However, the influence of organic solvents on the preparation of GQDs is still unknown. Herein, a systematic study on the carbonization of organic solvents toward GQDs is reported. The results show that organic solvents with the double bond or benzene ring or double hydrophilic groups could be directly decomposed into GQDs without the addition of catalysts or molecular precursors. The as-synthesized GQDs demonstrate ultra-small size distribution, high stability, tunable excitation wavelength and upconverted fluorescence. Both hematological and histopathological analyses show that the as-synthesized GQDs demonstrate a very good safety profile and excellent biocompatibility. The versatility of this synthesis strategy offers easy control of the surface group, composition, and optical properties of GQDs at the molecular level.


Eye Care Service Use and Its Determinants in Marginalized Communities in Pakistan: The Karachi Marine Fishing Communities Eye and General Health Survey.
To examine gender, ethnic, and socioeconomic differences in access to eye care services in marine fishing communities in Karachi, Pakistan. The Karachi Marine Fishing Communities Eye and General Health Survey was a door-to-door, cross-sectional survey conducted between March 2009 and April 2010 in fishing communities in Keamari, Karachi, located on the coast of the Arabian Sea. Adults aged ≥50 years living on three islands and in four coastal areas were enrolled. Participants underwent a detailed interview regarding sociodemographics, eye problems and eye care service use, testing of presenting and best-corrected visual acuity with a reduced logMAR chart, and detailed eye examination. A total of 700 people were planned to be included in the study; 638 (91.1%) were interviewed and examined. Most participants were extremely poor and had no formal education. Only 45.3% (95% confidence interval, CI, 41.4-49.2%) of participants reported having had an eye examination in the past; 12.1% (95% CI 9.5-14.6%) and 30.9% (95% CI 27.3-34.5%) had seen an eye doctor within the last year or prior 5 years, respectively. In the multivariable analysis, ethnicity was the strongest independent predictor of eye care service use, followed by self-reported eye problems and diabetes. Ethnic Bengalis were 4.2 times less likely (adjusted odds ratio 0.24, 95% CI 0.15-0.38; p < 0.001) to have had an eye examination in the past than Kutchis. Despite a high prevalence of visual impairment and blindness, levels of eye care examinations in fishing communities, especially among ethnic Bengalis, are disappointingly low. Such communities deserve particular attention in Vision 2020 and other national and international strategies and plans.


Interplay between the alpharetroviral Gag protein and SR proteins SF2 and SC35 in the nucleus.
Retroviruses are positive-sense, single-stranded RNA viruses that reverse transcribe their RNA genomes into double-stranded DNA for integration into the host cell chromosome. The integrated provirus is used as a template for the transcription of viral RNA. The full-length viral RNA can be used for the translation of the Gag and Gag-Pol structural proteins or as the genomic RNA (gRNA) for encapsidation into new virions by the Gag protein. The mechanism by which Gag selectively incorporates unspliced gRNA into virus particles is poorly understood. Although Gag was previously thought to localize exclusively to the cytoplasm and plasma membrane where particles are released, we found that the Gag protein of Rous sarcoma virus, an alpharetrovirus, undergoes transient nuclear trafficking. When the nuclear export signal of RSV Gag is mutated (Gag.L219A), the protein accumulates in discrete subnuclear foci reminiscent of nuclear bodies such as splicing speckles, paraspeckles, and PML bodies. In this report, we observed that RSV Gag.L219A foci appeared to be tethered in the nucleus, partially co-localizing with the splicing speckle components SC35 and SF2. Overexpression of SC35 increased the number of Gag.L219A nucleoplasmic foci, suggesting that SC35 may facilitate the formation of Gag foci. We previously reported that RSV Gag nuclear trafficking is required for efficient gRNA packaging. Together with the data presented herein, our findings raise the intriguing hypothesis that RSV Gag may co-opt splicing factors to localize near transcription sites. Because splicing occurs co-transcriptionally, we speculate that this mechanism could allow Gag to associate with unspliced viral RNA shortly after its transcription initiation in the nucleus, before the viral RNA can be spliced or exported from the nucleus as an mRNA template.


CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses.
Intracellular calcium acts as a secondary messenger in a wide variety of crucial biological signaling processes. Advances in fluorescence microscopy and calcium sensitive dyes has led to the routine quantification of calcium responses in non-excitable cells. However, the automatization of global intracellular calcium analysis at the single-cell level within a large population simultaneously remains challenging. One software, CalQuo (Calcium Quantification), offers some automatic features in calcium analysis. Here, we present an advanced version of the software package: CalQuo 2 . CalQuo 2 analyzes the calcium response in the Fourier-domain, allowing the number of user-defined filtering parameters to be reduced to one and a greater diversity of calcium responses to be recognized, compared to CalQuo that directly interprets the calcium intensity signal. CalQuo 2 differentiates cells that release a single calcium response and those that release oscillatory calcium fluxes. We have demonstrated the use of CalQuo 2 by measuring the calcium response in genetically modified Jurkat T-cells under varying ligand conditions, in which we show that peptide:MHCs and anti-CD3 antibodies trigger a fraction of T cells to release oscillatory calcium fluxes that increase with increasing koff rates. These results show that CalQuo 2 is a robust and user-friendly tool for characterizing global, single cell calcium responses.


Survival advantage of black patients with kidney disease after acute myocardial infarction.
Black individuals have a disproportionate incidence of ESRD when compared with white individuals, and among patients with ESRD, black patients experience better survival. The aim of this analysis is to assess, in a nationally representative sample of patients with cardiovascular disease, ethnic differences in survival among predialysis patients with kidney disease. A retrospective cohort analysis was conducted of Cooperative Cardiovascular Project data of Medicare patients who were aged > 65 yr and admitted for incident acute myocardial infarction and had 3 yr of mortality follow-up. Cox regression models and Kaplan Meier estimates were performed to examine differences in survival between black and white patients stratified by severity of kidney disease. Of 57,942 patients, 7.3% were black. Black patients were younger and more likely to be female and were less likely to have decreased kidney function. A significant interaction between race and kidney function existed with respect to mortality among patients who survived to discharge. The adjusted hazard ratios for death, black compared with white patients, were 1.00 (95% confidence interval 0.90 to 1.11) among patients with a GFR > or = 60 ml/min per 1.73 m2 and decreased monotonically among patients with lower GFR to 0.79 (95% confidence interval 0.61 to 0.97) among patients with a GFR 15 to 29 ml/min per 1.73 m2. Among patients with incident acute myocardial infarction, black patients with more severe kidney disease, when compared with their white counterparts, experience better survival. Further investigation into the reasons for ethnic differences in survival and progression of kidney disease is warranted.


Inactivation of D-alanine carboxypeptidase by penicillins and cephalosporins is not lethal in Bacillus subtilis.
The D-alanine carboxypeptidase of Bacillus subtilis is a particulate enzyme that is irreversibly inactivated by penicillins and cephalosporins. However, the lethal concentrations of these antibiotics are not the same as those that inhibit enzymatic activity in vitro. 6-Aminopenicillanic acid inactivates at least 95% of the enzyme at nonlethal concentrations. Conversely, cephalothin is lethal at concentrations that do not inactivate the enzyme. Experiments with intact, growing cells confirm the results obtained in vitro. Therefore, a killing site distinct from the carboxypeptidase must be postulated.


Melatonin is a biomarker of circadian dysregulation and is correlated with major depression and fibromyalgia symptom severity.
This study compared urinary 6-sulfatoxymelatonin (aMT6s) over 24 hours among fibromyalgia (FM), major depression disorder (MDD), and healthy control (HC) groups, and examined whether rhythm is correlated with depressive symptoms. To answer this question we compared the rhythm of urinary aMT6s secretion among each group in four time series: morning (06:00-12:00 hours), afternoon (12:00-18:00 hours), evening (18:00-24:00 hours), and night (24:00-06:00 hours). In the FM subjects, we assessed if the rhythm of urinary aMT6s secretion is associated with pain severity, sleep quality, number of trigger points (NTPs), and the pain pressure threshold (PPT). We included 54 women, aged 18-60 years with diagnosis of FM (n=18), MDD (n=19), and HC (n =17). The 24-hour urinary aMT6s was evaluated according to four standardized periods. The assessment instruments were the Hamilton Depression Rating Scale (HDRS), Pittsburgh Sleep Quality Index, and Fibromyalgia Impact Questionnaire. A generalized estimating equation revealed no difference in the daily load of aMT6s secretion among the three groups (P=0.49). However, at the daily time (06:00-18:00 hours), the load secretion of aMT6s reached 41.54% and 60.71% in the FM and MDD, respectively, as compared to 20.73% in the HC (P<0.05). A higher score in the HDRS was positively correlated with the amount of aMT6s secretion during daytime (06:00-18:00 hours). Also, multivariate linear regression revealed that in FM subjects, the aMT6s secretion during daytime (06:00-18:00 hours) was negatively correlated with the PPTlog (partial η2=0.531, P=0.001). However, it was positively correlated with depressive symptoms (partial η2=0.317, P=0.01); PQSI (partial η2=0.306, P=0.017), and NTPs (partial η2=0.23, P=0.04). A more significant load of aMT6s secretion during daytime hours was observed in MDD and FM subjects compared to HC. These findings help to comprehend the biological basis of these disorders and show how disruption in melatonin secretion is positively correlated with clinical symptoms.


Phase-shift tinnitus treatment: an open prospective clinical trial.
We report on a novel treatment for tinnitus using phase-shift pure tone sound treatment in patients with predominantly pure tone tinnitus. Thirty-five patients with pure tone tinnitus unresponsive to all previous treatment were enrolled in the study. All patients were treated three times in one week. If the patient noticed an improvement, the therapy was continued for six weeks with a home device customised to their specific treatment frequency. Twenty-one of the 35 patients (60%) responded positively to the initial therapy sessions. Tinnitus was assessed before treatment, after three in-office Tinnitus Phase-Out System therapy sessions, and after six weeks of home use of the Patient Treatment Device. The assessment instruments were a VAS loudness scale and the quality of life Tinnitus Questionnaire. Significant tinnitus reduction was obtained on VAS after three office Tinnitus Phase-Out System therapy sessions (before treatment: mean VAS = 6.4; after three therapy sessions: mean VAS = 4.9; p = 0.042) and after six weeks of home use of the Patient Treatment Device (mean VAS = 4.9; p = 0.005). When analysing the mean TQ score over treatment, there was a significant improvement in total score from pretreatment (mean TQ score = 41.9) to six weeks after home use of the Patient Treatment Device use (mean TQ score = 36.4) (p = 0.003). In view of the results obtained, the Phase-Out Treatment for tinnitus may provide the majority of patients with a significant improvement in their symptoms. Further evaluation, comparing this specific Phase-Out Treatment with more general noise stimulation treatment, will further specify the indications for this treatment option.


The effects of hepatocyte stimulating factor on fibrinogen biosynthesis in hepatocyte monolayers.
The biosynthesis of fibrinogen increased at least eightfold in primary hepatocytes when incubated in the presence of monocyte/macrophage-derived hepatocyte stimulating factor (HSF). The large increase in fibrinogen production is due to increased availability of the mRNAs for the protein since cytodot analysis of cellular RNA showed a 10-12-fold increase in each of the fibrinogen mRNAs. Pulse-chase experiments showed that the time for fibrinogen synthesis, assembly, and secretion was 40-50 min for both control and stimulating conditions. This indicates that the increased production was due principally to the presence of greater amounts of fibrinogen mRNA rather than translation or secretion-specific events. Three lines of evidence indicate that the increase in fibrinogen production was due to HSF effects on transcription: (a) analysis of cytoplasmic levels of each of the fibrinogen mRNAs showed that all three increased at the same rate and to the same extent, demonstrating that HSF affects the three gene products coordinately; (b) Northern gel analysis of cytoplasmic RNA isolated after very brief exposures to HSF showed increases in a large molecular weight fibrinogen RNA precursor; and (c) actinomycin D blocked the HSF-stimulated increase in fibrinogen mRNA species. Furthermore, experiments in which protein synthesis was inhibited by cycloheximide failed to inhibit the increase in fibrinogen mRNAs, indicating new protein synthesis is not required for the HSF stimulation of fibrinogen mRNA. These results are consistent with our hypothesis that HSF is exerting its control of fibrinogen at the level of gene transcription.


Analysis of photonic band structure in a one-dimensional photonic crystal containing single-negative materials.
A theoretical analysis on the angle- and thickness-dependent photonic band structure in a one-dimensional photonic crystal containing single-negative (SNG) materials is presented. The photonic crystal consists of two alternating SNG materials, including that one has a negative permittivity (ENG) and the other has a negative permeability (MNG). It is found that there are two types of SNG gaps. The first is the low-frequency gap which is very insensitive to the incident angle in the transversal electric (TE) wave. The second gap, which strongly relies on the incident angle for both TE and transversal magnetic (TM) waves, will close at the zero bandgap frequency at which the impedance match as well as the phase match in the constituent ENG and MNG layers must be simultaneously satisfied. This zero bandgap frequency is also strongly dependent on the incident angle. The band edges and the gap maps are investigated rigorously as a function of the incident angle and the ratio of thickness of the two SNG layers. The analyses are made in the lossless and lossy cases for both TE and TM waves. The inclusion of the loss enables us to further clarify two fundamentally distinct second SNG gaps which are separated by a threshold angle of incidence.


Life cycle assessment of refractory waste management in a Spanish steel works.
A life cycle assessment (LCA) is conducted on the current refractory waste management practices in a steel works in Spain producing around 6,000 tonnes of refractory waste in 2018. Management practices included direct reuse of spent magnesia-carbon (MgO-C) bricks, recycling of MgO-C bricks and high-alumina refractories by an external contractor and landfilling of monolithics and isostatic refractories, for which there were yet no established valorization routes. This current situation was compared to a hypothetical scenario where all refractories were disposed in landfill. The LCA included waste management activities (collection, transport, treatment) as well as production of substituted primary materials (dead-burned magnesia, calcined bauxite and new MgO-C bricks) produced in China and Germany. Results are discussed for four indicators: greenhouse-gas emissions, Non-renewable energy demand, land use and water use. Overall, per tonne of produced waste, the current management leads to a reduction or saving of 0.54 tonnes CO2-eq, 3 GJ primary energy from non-renewable sources, the occupation of 10 m2 of land during one year and the abstraction of 5 m3 freshwater. The results also show that reuse, in the particular case of spent MgO-C bricks, leads to higher benefits than recycling by At least a factor four. The robustness of these results is confirmed through the application of a Monte Carlo uncertainty analysis, as well as a sensitivity analysis focusing on a shift from coal to natural gas in the Chinese refractory industry.


Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time.
We have developed a method to measure binding of adenine nucleotides to intact, functional transmembrane receptors in a cellular or membrane environment. This method combines expression of proteins tagged with the fluorescent non-canonical amino acid ANAP, and FRET between ANAP and fluorescent (trinitrophenyl) nucleotide derivatives. We present examples of nucleotide binding to ANAP-tagged KATP ion channels measured in unroofed plasma membranes and excised, inside-out membrane patches under voltage clamp. The latter allows for simultaneous measurements of ligand binding and channel current, a direct readout of protein function. Data treatment and analysis are discussed extensively, along with potential pitfalls and artefacts. This method provides rich mechanistic insights into the ligand-dependent gating of KATP channels and can readily be adapted to the study of other nucleotide-regulated proteins or any receptor for which a suitable fluorescent ligand can be identified.


Formation and prevention of fractures in sol-gel-derived thin films.
Sol-gel-derived thin films play an important role as the functional coatings for various applications that require crack-free films to fully function. However, the fast drying process of a standard sol-gel coating often induces mechanical stresses, which may fracture the thin films. An experimental study on the crack formation in sol-gel-derived silica and organosilica ultrathin (submicron) films is presented. The relationships among the crack density, inter-crack spacing, and film thickness were investigated by combining direct micrograph analysis with spectroscopic ellipsometry. It is found that silica thin films are more prone to fracturing than organosilica films and have a critical film thickness of 300 nm, above which the film fractures. In contrast, the organosilica films can be formed without cracks in the experimentally explored regime of film thickness up to at least 1250 nm. These results confirm that ultrathin organosilica coatings are a robust silica substitute for a wide range of applications.


Effects of aerobic exercise on premenopausal sex hormone levels: results of the WISER study, a randomized clinical trial in healthy, sedentary, eumenorrheic women.
It is hypothesized that exercise can lead to a decrease in breast cancer risk through several hormonal and nonhormonal mechanisms. The WISER (Women In Steady Exercise Research) study investigated the effects of aerobic exercise on premenopausal sex hormone levels. Three hundred ninety-one sedentary, healthy, young eumenorrheic women were randomized either into an exercise intervention of 30 minutes of aerobic exercise 5 times a week for approximately 16 weeks (n = 212) or into a control group (n = 179). Serum levels of estradiol, estrone sulfate, testosterone, and sex hormone-binding globulin (SHBG), all in the midfollicular phase, and of progesterone, in the midluteal phase, were measured at baseline and at the end of the 16-week period. Compared with the controls (n = 153), exercisers (n = 166) experienced significant increases in aerobic fitness, lean body mass, and decreases in percent body fat. There were no significant changes in body weight and menstrual cycle length between or within groups. Progesterone decreased significantly in exercisers; however, this reduction was similar to that of the control group. No significant changes between or within groups were found for any of the other sex hormones or SHBG. In premenopausal women, 16 weeks of 150 minutes per week of moderate aerobic exercise in young women did not significantly alter sex hormone or SHBG levels. Any favorable effects that moderate aerobic exercise without an associated weight change may have on breast cancer risk in premenopausal women are unlikely to be a consequence of changes in levels of sex hormones or SHBG.


EMERGE: A Randomized Phase II Study of the Antibody-Drug Conjugate Glembatumumab Vedotin in Advanced Glycoprotein NMB-Expressing Breast Cancer.
Glycoprotein NMB (gpNMB), a negative prognostic marker, is overexpressed in multiple tumor types. Glembatumumab vedotin is a gpNMB-specific monoclonal antibody conjugated to the potent cytotoxin monomethyl auristatin E. This phase II study investigated the activity of glembatumumab vedotin in advanced breast cancer by gpNMB expression. Patients (n = 124) with refractory breast cancer that expressed gpNMB in ≥ 5% of epithelial or stromal cells by central immunohistochemistry were stratified by gpNMB expression (tumor, low stromal intensity, high stromal intensity) and were randomly assigned 2:1 to glembatumumab vedotin (n = 83) or investigator's choice (IC) chemotherapy (n = 41). The study was powered to detect overall objective response rate (ORR) in the glembatumumab vedotin arm between 10% (null) and 22.5% (alternative hypothesis) with preplanned investigation of activity by gpNMB distribution and/or intensity (Stratum 1 to Stratum 3). Glembatumumab vedotin was well tolerated as compared with IC chemotherapy (less hematologic toxicity; more rash, pruritus, neuropathy, and alopecia). ORR was 6% (five of 83) for glembatumumab vedotin versus 7% (three of 41) for IC, without significant intertreatment differences for predefined strata. Secondary end point revealed ORR of 12% (10 of 83) versus 12% (five of 41) overall, and 30% (seven of 23) versus 9% (one of 11) for gpNMB overexpression (≥ 25% of tumor cells). Unplanned analysis showed ORR of 18% (five of 28) versus 0% (0 of 11) in patients with triple-negative breast cancer (TNBC), and 40% (four of 10) versus 0% (zero of six) in gpNMB-overexpressing TNBC. Glembatumumab vedotin is well tolerated in heavily pretreated patients with breast cancer. Although the primary end point in advanced gpNMB-expressing breast cancer was not met for all enrolled patients (median tumor gpNMB expression, 5%), activity may be enhanced in patients with gpNMB-overexpressing tumors and/or TNBC. A pivotal phase II trial (METRIC [Metastatic Triple-Negative Breast Cancer]) is underway.


A Glimpse of the Peptide Profile Presentation by Xenopus laevis MHC Class I: Crystal Structure of pXela-UAA Reveals a Distinct Peptide-Binding Groove.
The African clawed frog, Xenopus laevis, is a model species for amphibians. Before metamorphosis, tadpoles do not efficiently express the single classical MHC class I (MHC-I) molecule Xela-UAA, but after metamorphosis, adults express this molecule in abundance. To elucidate the Ag-presenting mechanism of Xela-UAA, in this study, the Xela-UAA structure complex (pXela-UAAg) bound with a peptide from a synthetic random peptide library was determined. The amino acid homology between the Xela-UAA and MHC-I sequences of different species is <45%, and these differences are fully reflected in the three-dimensional structure of pXela-UAAg. Because of polymorphisms and interspecific differences in amino acid sequences, pXela-UAAg forms a distinct peptide-binding groove and presents a unique peptide profile. The most important feature of pXela-UAAg is the two-amino acid insertion in the α2-helical region, which forms a protrusion of ∼3.8 Å that is involved in TCR docking. Comparison of peptide-MHC-I complex (pMHC-I) structures showed that only four amino acids in β2-microglobulin that were bound to MHC-I are conserved in almost all jawed vertebrates, and the most unique feature in nonmammalian pMHC-I molecules is that the AB loop bound β2-microglobulin. Additionally, the binding distance between pMHC-I and CD8 molecules in nonmammals is different from that in mammals. These unique features of pXela-UAAg provide enhanced knowledge of T cell immunity and bridge the knowledge gap regarding the coevolutionary progression of the MHC-I complex from aquatic to terrestrial species.


Bacterial transformations of naphthothiophenes.
Naphthothiophenes are minor components of fossil fuels, and they can enter the environment from oil spills. Naphtho[2,1-b]thiophene, naphtho[2,3-b]thiophene, and 1-methylnaphtho[2,1-b]thiophene were synthesized and used in biodegradation studies with 1-methylnaphthalene (1-MN)-degrading Pseudomonas strains W1, F, and BT1. Cultures were incubated with one of the naphthothiophenes with or without 1-MN, acidified, and extracted with CH(inf2)Cl(inf2). The extracts were analyzed by gas chromatography with flame photometric and mass detectors to characterize sulfur-containing metabolites and with an atomic emission detector for quantification. Only strain W1 was able to grow on naphtho[2,1-b]thiophene, but strains F and BT1 cometabolized this compound if 1-MN was present. 1-MN was required by all three strains to metabolize naphtho[2,3-b]thiophene, which was more resistant to biodegradation than the [2,1-b] isomer. Two metabolites of naphtho [2,1-b]thiophene were purified, analyzed by (sup1)H nuclear magnetic resonance spectroscopy, and found to be 4-hydroxybenzothiophene-5-carboxylic acid (metabolite I) and 5-hydroxybenzothiophene-4-carboxylic acid (metabolite II). In cultures of strain W1 grown for 7 days on 52 (mu)mol of naphtho[2,1-b]thiophene, >84% of the substrate was degraded and metabolites I and II accounted for 19 and 9%, respectively, of the original amount of naphtho[2,1-b]thiophene. When 1-MN was present, strain W1 degraded >97% of the naphtho[2,1-b]thiophene and similar amounts of metabolite II were produced, but metabolite I did not accumulate. 1-MN was shown to promote the further degradation of metabolite I, but not of metabolite II, by strain W1. Thus, 1-MN enhanced the biodegradation of naphtho[2,1-b]thiophene. Approximately 70% of the 1-methylnaphtho [2,1-b]thiophene added to cultures of strain W1 with 1-MN was recovered as 4-hydroxy-3-methylbenzothiophene-5-carboxylic acid, the 3-methyl analog of metabolite I. The methyl substitution hindered further metabolism of 3-methyl-metabolite I even in the presence of 1-MN. Cometabolism of naphtho[2,3-b]thiophene yielded two products that were tentatively identified as 5-hydroxybenzothiophene-6-carboxylic and 6-hydroxybenzothiophene-5-carboxylic acids.


Single-molecule tracking technologies for quantifying the dynamics of gene regulation in cells, tissue and embryos.
For decades, we have relied on population and time-averaged snapshots of dynamic molecular scale events to understand how genes are regulated during development and beyond. The advent of techniques to observe single-molecule kinetics in increasingly endogenous contexts, progressing from in vitro studies to living embryos, has revealed how much we have missed. Here, we provide an accessible overview of the rapidly expanding family of technologies for single-molecule tracking (SMT), with the goal of enabling the reader to critically analyse single-molecule studies, as well as to inspire the application of SMT to their own work. We start by overviewing the basics of and motivation for SMT experiments, and the trade-offs involved when optimizing parameters. We then cover key technologies, including fluorescent labelling, excitation and detection optics, localization and tracking algorithms, and data analysis. Finally, we provide a summary of selected recent applications of SMT to study the dynamics of gene regulation.


Overlapping signals for protein degradation and nuclear localization define a role for intrinsic RAG-2 nuclear uptake in dividing cells.
Expression of the recombinase proteins RAG-1 and RAG-2 is discordant: while RAG-1 is relatively long lived, RAG-2 is degraded periodically at the G(1)-S transition. Destruction of RAG-2 is mediated by a conserved interval in the recombination-dispensable region. The need for RAG-2 to reaccumulate in the nucleus at each cell division suggested the existence of an intrinsic RAG-2 nuclear localization signal (NLS). RAG-1 or RAG-2, expressed individually, is a nuclear protein. A screen for proteins that bind the recombination-dispensable region of RAG-2 identified the nuclear transport protein Importin 5. Mutation of residues 499 to 508 in RAG-2 abolished Importin 5 binding, nuclear accumulation, and periodic degradation of RAG-2. The Importin 5 binding site overlaps an NLS, defined by mutagenesis. RAG-1 rescued the localization of degradation-defective, RAG-2 NLS mutants; this required an intact RAG-1 NLS. Mutations in RAG-2 that abolish intrinsic nuclear accumulation but spare periodic degradation impaired recombination in cycling cells; induction of quiescence restored recombination to wild-type levels. Recombination defects were correlated with a cell cycle-dependent defect in the ability of RAG-1 to rescue localization of the RAG-2 mutants. These results suggest that the intrinsic RAG-2 NLS functions in the nuclear uptake of RAG-2 following its reexpression in cycling cells.


Nuclear myocardial perfusion imaging with a cadmium-zinc-telluride detector technique: optimized protocol for scan time reduction.
We aimed at establishing the optimal scan time for nuclear myocardial perfusion imaging (MPI) on an ultrafast cardiac gamma-camera using a novel cadmium-zinc-telluride (CZT) solid-state detector technology. Twenty patients (17 male; BMI range, 21.7-35.5 kg/m(2)) underwent 1-d (99m)Tc-tetrofosmin adenosine stress and rest MPI protocols, each with a 15-min acquisition on a standard dual-detector SPECT camera. All scans were immediately repeated on an ultrafast CZT camera over a 6-min acquisition time and reconstructed from list-mode raw data to obtain scan durations of 1 min, 2 min, etc., up to a maximum of 6 min. For each of the scan durations, the segmental tracer uptake value (percentage of maximum myocardial uptake) from the CZT camera was compared by intraclass correlation with standard SPECT camera data using a 20-segment model, and clinical agreement was assessed per coronary territory. Scan durations above which no further relevant improvement in uptake correlation was found were defined as minimal required scan times, for which Bland-Altman limits of agreement were calculated. Minimal required scan times were 3 min for low dose (r = 0.81; P < 0.001; Bland-Altman, -11.4% to 12.2%) and 2 min for high dose (r = 0.80; P < 0.001; Bland-Altman, -7.6% to 12.9%), yielding a clinical agreement of 95% and 97%, respectively. We have established the minimal scan time for a CZT solid-state detector system, which allows 1-d stress/rest MPI with a substantially reduced acquisition time resulting in excellent agreement with regard to uptake and clinical findings, compared with MPI from a standard dual-head SPECT gamma-camera.


Applications of ALD MnO to electrochemical water splitting.
Atomic layer deposition (ALD) is an attractive method to deposit uniform catalytic films onto high surface area electrodes. One interesting material for ALD synthesis is MnOx, a promising earth-abundant catalyst for the oxygen evolution reaction (OER). It has previously been shown that catalysts beginning as MnO synthesized using ALD on smooth glassy carbon (s-GC) electrodes and Mn2O3 obtained upon annealing MnO on s-GC are active OER catalysts. Here, we use ALD to deposit MnO on high surface area GC (HSA-GC) substrates, forming an active catalyst on a geometric surface area basis. We then characterize three types of catalysts, HSA-GC MnO, s-GC MnO, and annealed MnO (Mn2O3), using cyclic voltammetry (CV), scanning electron microscopy (SEM), and ex situ X-ray absorption spectroscopy (XAS). We show that under OER conditions, all three catalysts oxidize to similar surface states with a mixture of Mn(3+)/Mn(4+) and that MnOx surface area effects can account for the observed differences in the catalytic activity. We also demonstrate the need for a high surface area support for high OER activity on a geometric basis.


Prevalence, comorbidity and heritability of hoarding symptoms in adolescence: a population based twin study in 15-year olds.
Hoarding Disorder (HD) is often assumed to be an 'old age' problem, but many individuals diagnosed with HD retrospectively report first experiencing symptoms in childhood or adolescence. We examined the prevalence, comorbidity and etiology of hoarding symptoms in adolescence. To determine the presence of clinically significant hoarding symptoms, a population-based sample of 15-year old twins (N = 3,974) completed the Hoarding Rating Scale-Self Report. Co-occurring Obsessive Compulsive Disorder (OCD), Autism Spectrum Disorders (ASD) and Attention Deficit Hyperactivity Disorder (ADHD) were estimated from parental report. Model-fitting analyses divided hoarding symptom scores into additive genetic, shared, and non-shared environmental effects. The prevalence of clinically significant hoarding symptoms was 2% (95% CI 1.6-2.5%), with a significantly higher prevalence in girls than boys. Exclusion of the clutter criterion (as adolescents do not have control over their environment) increased the prevalence rate to 3.7% (95% CI 3.1-4.3%). Excessive acquisition was reported by 30-40% among those with clinically significant hoarding symptoms. The prevalence of co-occurring OCD (2.9%), ASD (2.9%) and ADHD (10.0%) was comparable in hoarding and non-hoarding teenagers. Model-fitting analyses suggested that, in boys, additive genetic (32%; 95% CI 13-44%) and non-shared environmental effects accounted for most of the variance. In contrast, among girls, shared and non-shared environmental effects explained most of the variance, while additive genetic factors played a negligible role. Hoarding symptoms are relatively prevalent in adolescents, particularly in girls, and cause distress and/or impairment. Hoarding was rarely associated with other common neurodevelopmental disorders, supporting its DSM-5 status as an independent diagnosis. The relative importance of genetic and shared environmental factors for hoarding differed across sexes. The findings are suggestive of dynamic developmental genetic and environmental effects operating from adolescence onto adulthood.


SINUS STIMULATION AS A FACTOR IN THE RESUSCITATION OF THE HEART.
The four series of experiments described above indicate clearly that tetanic stimulation of the sinus region of the auricles is of material assistance to massage in the resuscitation of the heart. It causes the auricles to generate impulses, either rhythmical or irregular, to which the ventricles, when susceptible, respond. When massage administered for brief periods has failed to resuscitate the heart, the same period of massage combined with tetanic stimulation of the sinus has usually brought the circulation back to normal. In some instances massage alone when continued over long periods has been without effect, and to all appearances would have continued to be without effect, whereas at such a time massage combined with stimulation of the sinus has effected recovery. Whether the stimulus causes the auricles to beat normally or to fibrillate seems to make no essential difference in the end result. Not infrequently conductivity and irritability are lowered to such an extent by the cessation of the circulation, that impulses generated in an auricle started by stimulation can at first manifest no effect upon the ventricles. Under such circumstances massage alone or, more frequently, massage plus stimulation may cause the ventricles to beat with their inherent slow rate. Often then the force of the ventricular contractions is insufficient for the maintenance of an effective circulation. In such cases, however, with the gradual improvement in the condition of the heart, the auricular impulses, as a rule, soon begin to reach the ventricles, with the result that the latter immediately begin to beat more rapidly and effectively. The beneficial results of stimulation of the auricles, therefore, manifest themselves relatively late, but none the less effectively. It has been found that even after the heart has begun to beat, stimulation of the sinus region may still be of some assistance, though in another direction. It not infrequently happens that after resuscitation of the heart the blood pressure fails to rise above a certain low and insufficient level. In such cases repeated tetanic stimulation of the sinus may accelerate the rise of blood pressure and so hasten recovery (protocol of experiment 12, series II, and text-figure 5). No attempt has been made to determine the mechanism of this effect.


An Anacardiaceae preparation reduces the expression of inflammation-related genes in murine macrophages.
This study investigated the effects of an aqueous extract of the stem bark of Mangifera indica L. (Anacardiaceae; Vimang), which contains a defined mixture of components including polyphenols (principally mangiferin, MA), triterpenes, phytosteroids, fatty acids and microelements, on expression of inflammation mediators in inflammatory murine macrophages after stimulation in vitro with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). In vitro treatment with Vimang at 4 microg/ml reduced levels of NOS-2 mRNA and NOS-2, while treatment at 40 microg/ml also reduced levels of COX-2 mRNA, COX-2, and prostaglandin E2 (PGE2). Results suggested that MA is involved in these effects. In vitro treatment with Vimang at 40 microg/ml also inhibited mRNA levels of the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and colony-stimulating factor (GM-CSF), but did not affect mRNA levels of IL-6 or tumor growth factor-beta (TGF-beta). Extracellular release of TNF-alpha by inflammatory macrophages was inhibited by in vitro treatment with Vimang at the same concentrations that showed inhibition of TNF-alpha mRNA levels. The inhibition of TNF-alpha production appears to be at least partially attributable to MA. Vimang at 4 microg/ml decreased mRNA levels of nuclear factor-kappaB (NF-kappaB) but did not affect expression of the NF-kappaB inhibitor (IkappaB). These data indicate that the potent anti-inflammatory effects of Vimang are due to selective modulation of the expression of inflammation-related genes, leading to attenuation of macrophage activation.


Contrasted innate responses to two viruses in zebrafish: insights into the ancestral repertoire of vertebrate IFN-stimulated genes.
Ease of imaging and abundance of genetic tools make the zebrafish an attractive model host to understand host-pathogen interactions. However, basic knowledge regarding the identity of genes involved in antiviral immune responses is still lagging in this species. We conducted a microarray analysis of the larval zebrafish response to two models of RNA virus infections with very different outcomes. Chikungunya virus (CHIKV) induces a rapid and protective IFN response. Infection with infectious hematopoietic necrosis virus is lethal and is associated with a delayed and inefficient IFN response. A typical signature of IFN-stimulated genes (ISGs) was observed with both viruses, but was stronger for CHIKV. We further compared the zebrafish and human ISG repertoires and made a genomic and phylogenic characterization of the main gene families. We describe a core set of well-induced ISGs conserved across vertebrates, as well as multigenic families diversified independently in each taxon. The conservation of ISGs involved in antiviral signaling indicates conservation of the main feedback loops in these pathways. Whole-mount in situ hybridization of selected transcripts in infected larvae revealed a typical pattern of expression for ISGs in the liver, gut, and blood vessels with both viruses. We further show that some inflammatory genes were additionally induced through IFN-independent pathways by infectious hematopoietic necrosis virus and not by CHIKV. This study provides a useful reference set for the analysis of host-virus interactions in zebrafish and highlights the differences between protective and nonprotective antiviral innate responses.


Cross-Plane and In-Plane Heat Conductions in Layer-by-Layer Membrane: Molecular Dynamics Study.
A material with anisotropic heat conduction characteristics, which is determined by molecular scale structure, provides a way of controlling heat flow in nanoscale spaces. As such, here, we consider layer-by-layer (LbL) membranes, which are an electrostatic assembly of polyelectrolyte multilayers and are expected to have different heat conduction characteristics between cross-plane and in-plane directions. We constructed models of a poly(acrylic acid)/polyethylenimine (PAA/PEI) LbL membrane sandwiched by charged solid walls and investigated their anisotropic heat conduction using molecular dynamics simulations. In the cross-plane direction, the thermal boundary resistance between the solid wall and the LbL membrane and that between the constituent PAA and PEI layers decrease with increasing degree of ionization (solid surface charge density and the number of electric charges per PAA/PEI molecule). When the degree of ionization is low, the cross-plane thermal conductivity of a constituent layer is higher than that of the bulk state. As the degree of ionization increases, however, the cross-plane thermal conductivity of PAA, a linear polymer, decreases because of the increase in the number of in-plane oriented polymer chains. In the in-plane direction, we investigated the heat conduction of each layer and found the enhancement of effective in-plane thermal conductivity again due to the in-plane oriented chain alignment. The heat conduction in the LbL membrane is three-dimensionally enhanced compared to those in the bulk states of the constituent polymers because of the electrostatic interactions in the cross-plane direction and the molecular alignment in the in-plane direction.