The MEST score provides earlier risk prediction in lgA nephropathy.
The Oxford Classification of IgA nephropathy (IgAN) includes the following four histologic components: mesangial (M) and endocapillary (E) hypercellularity, segmental sclerosis (S) and interstitial fibrosis/tubular atrophy (T). These combine to form the MEST score and are independently associated with renal outcome. Current prediction and risk stratification in IgAN requires clinical data over 2 years of follow-up. Using modern prediction tools, we examined whether combining MEST with cross-sectional clinical data at biopsy provides earlier risk prediction in IgAN than current best methods that use 2 years of follow-up data. We used a cohort of 901 adults with IgAN from the Oxford derivation and North American validation studies and the VALIGA study followed for a median of 5.6 years to analyze the primary outcome (50% decrease in eGFR or ESRD) using Cox regression models. Covariates of clinical data at biopsy (eGFR, proteinuria, MAP) with or without MEST, and then 2-year clinical data alone (2-year average of proteinuria/MAP, eGFR at biopsy) were considered. There was significant improvement in prediction by adding MEST to clinical data at biopsy. The combination predicted the outcome as well as the 2-year clinical data alone, with comparable calibration curves. This effect did not change in subgroups treated or not with RAS blockade or immunosuppression. Thus, combining the MEST score with cross-sectional clinical data at biopsy provides earlier risk prediction in IgAN than our current best methods.


Plasma IL-1 Receptor Antagonist Concentration Has an Inverse Association With Birth Weight in Prepubertal Children.
Birth size has an impact on later cardiometabolic risk that is strongly related to low-grade inflammation. To evaluate plasma interleukin-1 receptor antagonist (IL-1ra) concentrations in relation to birth size and cardiometabolic and inflammatory markers in prepubertal children. A cohort study. Anthropometric data were recorded. Fasting blood samples were collected for plasma analyses of IL-1ra, alanine transaminase, total cholesterol, high- and low-density lipoprotein cholesterols, triglyceride, glucose, and serum analyses of 25-hydroxyvitamin D [25(OH)D] and high-sensitivity C-reactive protein (hs-CRP) concentrations. Forty-nine large for gestational age (LGA), 56 appropriate for gestational age, and 23 small for gestational age (SGA) children at 5 to 8 years of age were examined. Differences in IL-1ra concentrations among the birth-size groups and associations between IL-1ra and other metabolic markers were assessed. Body mass index (BMI) standard deviation score (SDS)-adjusted plasma IL-1ra concentrations were highest in the SGA- and lowest in the LGA-born children (P = 0.015). Age- and sex-adjusted IL-1ra concentrations had strongest associations with BMI SDS (P < 0.001) and hs-CRP (P < 0.001, also when further adjusted for BMI SDS). Prepubertal children born SGA had the highest and those born LGA the lowest IL-1ra concentrations in this study cohort. Most associations found between IL-1ra and the studied metabolic parameters were weight related, but the association with hs-CRP remained strong after adjustment for BMI. It seems that at prepuberty, SGA children have a stronger inflammatory state than LGA children and may thus be at a greater risk for later metabolic disturbances.


Molecular dynamics simulations of large integral membrane proteins with an implicit membrane model.
The heterogeneous dielectric generalized Born (HDGB) methodology is an the extension of the GBMV model for the simulation of integral membrane proteins with an implicit membrane environment. Three large integral membrane proteins, the bacteriorhodopsin monomer and trimer and the BtuCD protein, were simulated with the HDGB model in order to evaluate how well thermodynamic and dynamic properties are reproduced. Effects of the truncation of electrostatic interactions were examined. For all proteins, the HDGB model was able to generate stable trajectories that remained close to the starting experimental structures, in excellent agreement with explicit membrane simulations. Dynamic properties evaluated through a comparison of B-factors are also in good agreement with experiment and explicit membrane simulations. However, overall flexibility was slightly underestimated with the HDGB model unless a very large electrostatic cutoff is employed. Results with the HDGB model are further compared with equivalent simulations in implicit aqueous solvent, demonstrating that the membrane environment leads to more realistic simulations.


Near infrared two photon imaging using a bright cationic Yb(iii) bioprobe spontaneously internalized into live cells.
An Yb(iii) complex based on a dimethyl cyclen macrocyclic ligand functionalized by charge transfer antennae was prepared. This cationic [YbL3]+ complex is stable and soluble in water and presents interesting photophysical nonlinear properties. It is spontaneously internalized and accumulates in live cells. High quality images have been obtained both in a classical NIR-to-vis configuration and in the more challenging NIR-to-NIR one.


Insurance coverage of special foods needed in the treatment of phenylketonuria.
Optimal medical management of phenylketonuria (PKU) requires the use of special low-phenylalanine foods for many years. For women with PKU, elevated maternal blood levels of phenylalanine even at conception can lead to fetal damage. Despite this need, private health insurance, Medicaid, and other public health programs often exclude the cost of these foods from their benefits. The New York State Department of Health conducted a survey of metabolic disorders treatment centers to elucidate the problems PKU patients have obtaining and paying for the special foods essential to their care. Payment for special foods was denied to nearly half of those with private health insurance policies and was covered for only 10 percent of Medicaid-eligibles. A public program for children with special health care needs covered these food costs in upstate New York but not in New York City. There is no program of assistance for adults who are not eligible for Medicaid and who do not have private insurance coverage of special foods. At present, many private health insurance policies and public programs do not cover the costs of low-phenylalanine foods other than infant formula. Payment for this essential part of the management of PKU should be mandated for all public programs for persons with chronic illnesses, public medical assistance (Medicaid) programs, and private health insurance. There is a need for a public program to assist adults with PKU who are not eligible for Medicaid and who do not have health insurance that covers these costs.


A giant planet candidate transiting a white dwarf.
Astronomers have discovered thousands of planets outside the Solar System1, most of which orbit stars that will eventually evolve into red giants and then into white dwarfs. During the red giant phase, any close-orbiting planets will be engulfed by the star2, but more distant planets can survive this phase and remain in orbit around the white dwarf3,4. Some white dwarfs show evidence for rocky material floating in their atmospheres5, in warm debris disks6-9 or orbiting very closely10-12, which has been interpreted as the debris of rocky planets that were scattered inwards and tidally disrupted13. Recently, the discovery of a gaseous debris disk with a composition similar to that of ice giant planets14 demonstrated that massive planets might also find their way into tight orbits around white dwarfs, but it is unclear whether these planets can survive the journey. So far, no intact planets have been detected in close orbits around white dwarfs. Here we report the observation of a giant planet candidate transiting the white dwarf WD 1856+534 (TIC 267574918) every 1.4 days. We observed and modelled the periodic dimming of the white dwarf caused by the planet candidate passing in front of the star in its orbit. The planet candidate is roughly the same size as Jupiter and is no more than 14 times as massive (with 95 per cent confidence). Other cases of white dwarfs with close brown dwarf or stellar companions are explained as the consequence of common-envelope evolution, wherein the original orbit is enveloped during the red giant phase and shrinks owing to friction. In this case, however, the long orbital period (compared with other white dwarfs with close brown dwarf or stellar companions) and low mass of the planet candidate make common-envelope evolution less likely. Instead, our findings for the WD 1856+534 system indicate that giant planets can be scattered into tight orbits without being tidally disrupted, motivating the search for smaller transiting planets around white dwarfs.


Magnesium supplementation in the prevention of arrhythmias in pediatric patients undergoing surgery for congenital heart defects.
The efficacy of magnesium in the prevention of arrhythmias in pediatric patients after heart surgery remains unknown. Therefore we prospectively examined the effect of magnesium treatment on the incidence of postoperative arrhythmias in pediatric patients undergoing surgical repair of congenital heart defects. Twenty-eight pediatric patients undergoing heart surgery with cardiopulmonary bypass were prospectively, randomly assigned in a double-blind fashion to receive intravenous magnesium (magnesium group, n = 13; 30 mg/kg) or saline (placebo group, n = 15) immediately after cessation of cardiopulmonary bypass. Magnesium, potassium, and calcium levels were measured at defined intervals during surgery and 24 hours after surgery. Continuous electrocardiographic documentation by Holter monitor was performed for 24 hours after surgery. Magnesium levels were significantly decreased below the normal reference range for patients in the placebo group compared with the magnesium group on arrival in the intensive care unit and for 20 hours after surgery. Magnesium levels remained in the normal range for patients in the magnesium group after magnesium supplementation. In 4 patients in the placebo group (27%), junctional ectopic tachycardia developed within the initial 20 hours in the intensive care unit. No junctional ectopic tachycardia was observed in the magnesium group (P =.026). Although this study was originally targeted to include 100 patients, the protocol was terminated because of the unacceptable incidence of hemodynamically significant junctional ectopic tachycardia that was present in the placebo group. Thus low magnesium levels in pediatric patients undergoing heart surgery are associated with an increased incidence of junctional ectopic tachycardia in the immediate postoperative period.


Gestational diabetes in IVF and spontaneous pregnancies.
Increasing number of pregnancies resulting from In vitro Fertilisation (IVF) combined with increased awareness of Gestational Diabetes Mellitus (GDM) - related morbidity calls for research on possible differences in pregnancy outcomes between IVF and non-IVF GDM complicated pregnancies. The aim of this study was to compare GDM - affected pregnancies, both resulting from IVF and conceived without medical intervention with regards to maternal data, fetal biometry and neoanatal outcomes. We used our Clinic's databases to retrospectively identify 36 women who had successful IVF and developed GDM during the course of their singleton pregnancy (IVF group) and 137 non-IVF women with GDM (non-IVF group). They were matched according to age, pre-pregnancy BMI and none had the history of diabetes mellitus before their pregnancies. We compared the maternal characteristics, course of pregnancy and neonatal outcome. The weight gain until the diagnosis of GDM in both non-IVF and IVF groups of women was not significantly different (9.81±4.37 vs 10.0±4.8 kg, p=0.8 respectively) with similar time at which they came under the specialist GDM care (29±4.0 vs 28±4.5 wks, p=0.42,). When analyzing first trimester fasting glucose levels we found it to be significantly higher in IVF group (89±16.2 vs 83±11.3 mg/dl, p=0.04). Second trimester oral glucose tolerance test (OGTT) results and glucose levels during GDM treatment did not differ between the groups. No changes were noted in investigated fetal and neonatal variances: 3rd trimester AC, it's percentile and neonatal birth weight (3 460±641 vs 3 200±440 g, p=0.22). GDM among women after in vitro fertilisation is characterised by higher first trimester fasting glucose levels. Early diagnostic intervention in IVF pregnancies is specially needed.


Profile of circulating levels of IL-1Ra, CXCL10/IP-10, CCL4/MIP-1β and CCL2/MCP-1 in dengue fever and parvovirosis.
Dengue virus (DENV) and parvovirus B19 (B19V) infections are acute exanthematic febrile illnesses that are not easily differentiated on clinical grounds and affect the paediatric population. Patients with these acute exanthematic diseases were studied. Fever was more frequent in DENV than in B19V-infected patients. Arthritis/arthralgias with DENV infection were shown to be significantly more frequent in adults than in children. The circulating levels of interleukin (IL)-1 receptor antagonist (Ra), CXCL10/inducible protein-10 (IP-10), CCL4/macrophage inflammatory protein-1 beta and CCL2/monocyte chemotactic protein-1 (MCP-1) were determined by multiplex immunoassay in serum samples obtained from B19V (37) and DENV-infected (36) patients and from healthy individuals (7). Forward stepwise logistic regression analysis revealed that circulating CXCL10/IP-10 tends to be associated with DENV infection and that IL-1Ra was significantly associated with DENV infection. Similar analysis showed that circulating CCL2/MCP-1 tends to be associated with B19V infection. In dengue fever, increased circulating IL-1Ra may exert antipyretic actions in an effort to counteract the already increased concentrations of IL-1β, while CXCL10/IP-10 was confirmed as a strong pro-inflammatory marker. Recruitment of monocytes/macrophages and upregulation of the humoral immune response by CCL2/MCP-1 by B19V may be involved in the persistence of the infection. Children with B19V or DENV infections had levels of these cytokines similar to those of adult patients.


Ricoeur's "Petite éthique": an ethical epistemological perspective for clinician-bioethicists.
The passage from a posture of clinician to that of clinician-bioethicist poses significant challenges for health professionals, most notably with regards to theoretical or epistemological views of complex ethical impasses encountered in clinical settings. Apprehending these situations from the only clinical perspective of the nurse or the doctor, for example, can be very unproductive to help solve this kind of situation and certainly poses great limits to the role of the clinician-bioethicist. Drawing on my own experience as a former nurse who, following graduate studies in bioethics has begun providing ethics consultation services, I argue that clinicians must undergo an epistemological transformation in order to become clinician-bioethicists. A source of inspiration or framework for would-be clinician-bioethicists is, I suggest, the "Petite éthique" developed by the contemporary French philosopher Paul Ricoeur. Specifically, clinician-bioethicists should develop specific core ethical competencies (in line with the conclusions of the American Society for Bioethics and Humanities (Core competencies for health care ethics consultation, 1998); namely: savoir or knowing, savoir faire or knowing how to do, and savoir être or knowing how to be.


Premature pulmonary valve opening.
Premature opening of the pulmonary valve (opening independent of atrial or ventricular systole) was originally described in a case of sinus of Valsalva rupture into the right atrium. Since that time we have observed five additional cases in which the pulmonary valve opened prematurely. Entities encountered included: 1) constrictive pericarditis; 2) Loeffler's endocarditis; 3) Ebstein's anomaly with tricuspid regurgitation; 4) tricuspid regurgitation following tricuspid valvulectomy, and 5) pulmonary regurgitation accompanied by atrial septal defect. In the first two cases, premature pulmonary valve opening is felt to be due to restriction of diastolic filling of the right ventricle with subsequent early diastolic rise in pressure equalling or exceeding pulmonary artery diastolic pressure. In the latter three cases, the increased volume of blood entering the right ventricle again appeared to result in a rapid rise in initial right ventricular diastolic pressure and to produce premature opening of the pulmonary valve. Premature pulmonary valve opening, therefore, does not appear specific for any particular clinical entity but reflects the relative pressures in the right ventricle and pulmonary artery during diastole.


Contemporary Pediatric Palliative Care: Myths and Barriers to Integration into Clinical Care.
At least 20 million children would benefit from Pediatric Palliative Care (PPC) annually, and 8 million children would need specialized PPC services. In the USA alone, more than 42,000 children 0-19 years died in 2013, fifty-five percent of them infants younger than 1 year. This article aims to critically review eight common assumptions, myths and barriers, which may hinder the implementation of PPC into the care of a child with advanced serious illness. Interdisciplinary PPC is about matching treatment to patient goals and is considered specialized medical care for children with serious illness. It is focused on relieving pain, distressing symptoms, and stress of a serious illness and appropriate at any age and at any stage, together with curative treatment. The primary goal is to improve the quality of life of the child and his or her family. Emerging evidence shows, that palliative care involvement results in improved quality of life as well as prolongation of life.


Can hantavirus infections be predicted on admission to hospital?
The aim of this study was to investigate the predictive factors which contribute to diagnosis of hantavirus infection. One hundred patients from rural areas hospitalized with a preliminary diagnosis of hantavirus infection from different hospitals in Turkey were investigated. Hantavirus infection was confirmed in 20 patients (Group 1) using immunofluorescence and immunoblot assays at the Refik Saydam National Public Health Agency. Hantaviruses were not detected in the serum of the remaining 80 patients, other infectious and non-infectious diseases being diagnosed in this group (Group 2). Patients' demographic characteristics and clinical and laboratory data on admission were examined and compared between the two groups. Fever, proteinuria, hematuria, lethargy-weakness, and nausea-vomiting were the most frequent symptoms and findings in Group 1, seen in almost all patients. Proteinuria, hematuria, muscle pain, diarrhea/abdominal pain, hypotension, shock, and sweating were observed at significantly higher levels in Group 1 compared to Group 2. Serum urea, creatinine, uric acid, lactate dehydrogenase (LDH), aspartate transaminase (AST), alkaline phosphatase (ALP), and C-reactive protein (CRP) were significantly higher, but serum platelet counts were lower in Group 1 patients. Area beneath the receiver operating characteristics (ROC) curve analysis was used to calculate the discriminative ability of various laboratory values to identify patients with hantavirus infection. This analysis revealed that, serum CRP had a 100% negative predictive value, whilst, platelet, and creatinine had 75% and 70% positive predictive values for the diagnosis of hantavirus infection. In summary, laboratory markers used in clinical practice are of great importance predicting hantavirus infections.


Bilateral syringeal coupling during phonation of a songbird.
The syrinx of oscine birds ("true songbirds") is a double vocal organ, and each side has generally been presumed to function independently under separate neural control during phonation. A significant counterexample is demonstrated here in the production of a common vocalization by the black-capped chickadee (Parus atricapillus), in which the 2 acoustic sources of the syrinx interact in a nonlinear fashion. The chickadee produces a sound with multiple frequency components that superficially resemble harmonics. An analysis of vocal production after unilateral and bilateral syringeal denervation shows instead that these frequency components are sum and difference frequencies, or heterodyne frequencies, resulting from cross-modulation between the 2 syringeal sides. A limited form of this bilateral coupling may be achieved after unilateral denervation of either syringeal side but not after bilateral denervation. Unilaterally denervated birds are capable of significant improvement in coupling after 10 d, too short a period for neural regrowth. These results suggest that coupling arises from a passive physical interaction between the 2 syringeal sources which is activated or regulated in some fashion by neural control from either side.


Synergistic effects of zinc oxide nanoparticles and Fatty acids on toxicity to caco-2 cells.
Fatty acids exposure may increase sensitivity of intestinal epithelial cells to cytotoxic effects of zinc oxide (ZnO) nanoparticles (NPs). This study evaluated the synergistic effects of ZnO NPs and palmitic acid (PA) or free fatty acids (FFAs) mixture (oleic/PA 2:1) on toxicity to human colon epithelial (Caco-2) cells. The ZnO NPs exposure concentration dependently induced cytotoxicity to Caco-2 cells showing as reduced proliferation and activity measured by 3 different assays. PA exposure induced cytotoxicity, and coexposure to ZnO NPs and PA showed the largest cytotoxic effects. The presence of FFAs mixture did not affect the ZnO NPs-induced cytotoxicity. Filtration of freshly prepared suspension of NPs through a 0.45-µm pore size membrane significantly reduced the cytotoxicity, indicating a role of concentration or size of particles in cytotoxic effects. The ZnO NPs and PA coexposure induced production of mitochondrial reactive oxygen species (mROS) but not intracellular ROS production, whereas FFAs mixture exposure did not induce mROS and inhibited intracellular ROS. Both ZnO NPs and fatty acids (PA and FFAs mixture) promoted lysosomal destabilization, which was not correlated with cytotoxicity. These results indicated that PA can enhance ZnO NPs-induced cytotoxicity probably by the augmentation of mROS production, whereas FFAs mixture did not affect ROS production. Synergistic effects between ZnO NPs and fatty acids may be important when considering NPs toxicity via oral exposure.


The epidermal growth factor receptor decreases Stathmin 1 and triggers catagen entry in the mouse.
The epidermal growth factor receptor (EGFR) is necessary for normal involution of hair follicles after the growth phase of anagen, although the mechanisms through which it acts are not well understood. In this report, we used transcriptional profiling of microdissected hair follicles from mice with skin-targeted deletion of Egfr to investigate how EGFR activation triggers catagen. Immunofluorescence for phospho-EGFR in mouse skin revealed increased activation of EGFR in follicular keratinocytes at catagen onset. Consistent with other models of EGFR deficiency, mice with skin-targeted deletion of Egfr (Krt14-Cre(+) /Egfr(fl/fl) ) exhibited a delayed and asynchronous catagen entry. Transcriptional profiling at the time of normal catagen onset at post-natal day (P) 17 revealed increased expression of the mitotic regulator Rcc2 in hair follicles lacking EGFR. Rcc2 protein was strongly immunopositive in the nuclei of control follicular keratinocytes at P16 then rapidly decreased until it was undetectable between P18 and 21. In contrast, Rcc2 expression continued in Egfr mutant follicles throughout this period. Proliferation, measured by bromodeoxyuridine incorporation, was also significantly increased in Egfr mutant follicular keratinocytes compared to controls at P18-21. Similarly, Rcc2-regulated mitotic regulator Stathmin 1 was strikingly reduced in control but not Egfr mutant follicles between P17 and P19. Deletion of Stmn1, in turn, accelerated catagen entry associated with premature cessation of proliferation in the hair follicles. These data reveal EGFR suppression of mitotic regulators including Rcc2 and Stathmin 1 as a mechanism for catagen induction in mouse skin.


Translational animal models for Alzheimer's disease: An Alzheimer's Association Business Consortium Think Tank.
Over 5 million Americans and 50 million individuals worldwide are living with Alzheimer's disease (AD). The progressive dementia associated with AD currently has no cure. Although clinical trials in patients are ultimately required to find safe and effective drugs, animal models of AD permit the integration of brain pathologies with learning and memory deficits that are the first step in developing these new drugs. The purpose of the Alzheimer's Association Business Consortium Think Tank meeting was to address the unmet need to improve the discovery and successful development of Alzheimer's therapies. We hypothesize that positive responses to new therapies observed in validated models of AD will provide predictive evidence for positive responses to these same therapies in AD patients. To achieve this goal, we convened a meeting of experts to explore the current state of AD animal models, identify knowledge gaps, and recommend actions for development of next-generation models with better predictability. Among our findings, we all recognize that models reflecting only single aspects of AD pathogenesis do not mimic AD. Models or combinations of new models are needed that incorporate genetics with environmental interactions, timing of disease development, heterogeneous mechanisms and pathways, comorbidities, and other pathologies that lead to AD and related dementias. Selection of the best models requires us to address the following: (1) which animal species, strains, and genetic backgrounds are most appropriate; (2) which models permit efficient use throughout the drug development pipeline; (3) the translatability of behavioral-cognitive assays from animals to patients; and (4) how to match potential AD therapeutics with particular models. Best practice guidelines to improve reproducibility also need to be developed for consistent use of these models in different research settings. To enhance translational predictability, we discuss a multi-model evaluation strategy to de-risk the successful transition of pre-clinical drug assets to the clinic.


Effect of added fat on the plasma glucose and insulin response to ingested potato given in various combinations as two meals in normal individuals.
In normal subjects, ingestion of fat with potato in a morning meal resulted in a decrease in the glucose response. Therefore, we wished to determine whether a fat-induced decrease in blood glucose also would be observed after a second identical meal. In addition, we were interested in determining if fat ingestion with a morning meal would have an effect on the blood glucose and insulin responses to a second meal not containing fat. Nine healthy male subjects ingested two meals consisting of an amount of potato containing 50 g carbohydrate, either alone or with 50 g fat as butter. The meals were served in four combinations as follows: 1) potato for the first meal, potato for the second meal; 2) potato for the first meal, potato with fat for the second meal; 3) potato with fat for the first meal, potato for the second meal; and 4) potato with fat for the first meal, potato with fat for the second meal. Meals were ingested at 8:00 A.M. and noon. Plasma glucose and C-peptide, serum insulin, triglyceride, and free fatty acid (FFA) concentrations were determined over an 8-h period. The integrated area responses to the meals were quantified over the subsequent 4-h period using the fasting value or the noon value as baseline for the first and second meals, respectively. When the first meal contained potato only, the glucose area response to the second meal was significantly less when the second meal contained fat. However, fat ingestion had no effect on the glucose area response to the second meal when fat was present in the first meal. The insulin area responses to the first and second meals were similar after ingestion of potato or potato with fat. However, the insulin response to the second meal always was less than that to the first meal. The C-peptide area responses after ingestion of the second meal also were all higher than those after the first meal. The triglyceride area responses were slightly negative after ingestion of potato alone in the first meal. When fat was ingested, they were positive. When the first meal contained fat but the second meal did not, there was a rise in triglyceride concentration after the second meal as well as after the first meal. That is, a rise occurred without ingestion of fat with the second meal. If fat was present in the second meal the rise was even greater. The FFA area responses were similar to the triglyceride area responses. When fat was ingested with carbohydrate in either the first or second meal, the glucose area response was decreased. However, when both meals contained fat, a decrease in the glucose area response did not occur with the second meal. The glucose area responses all were greater after the second meal compared with those after the first meal, i.e., the opposite of a Staub-Traugott effect was observed. The insulin area responses to the first and second meals were similar whether fat was ingested or not.


Tumor-infiltrating myeloid cells induce tumor cell resistance to cytotoxic T cells in mice.
Cancer immunotherapeutic approaches induce tumor-specific immune responses, in particular CTL responses, in many patients treated. However, such approaches are clinically beneficial to only a few patients. We set out to investigate one possible explanation for the failure of CTLs to eliminate tumors, specifically, the concept that this failure is not dependent on inhibition of T cell function. In a previous study, we found that in mice, myeloid-derived suppressor cells (MDSCs) are a source of the free radical peroxynitrite (PNT). Here, we show that pre-treatment of mouse and human tumor cells with PNT or with MDSCs inhibits binding of processed peptides to tumor cell-associated MHC, and as a result, tumor cells become resistant to antigen-specific CTLs. This effect was abrogated in MDSCs treated with a PNT inhibitor. In a mouse model of tumor-associated inflammation in which the antitumor effects of antigen-specific CTLs are eradicated by expression of IL-1β in the tumor cells, we determined that therapeutic failure was not caused by more profound suppression of CTLs by IL-1β-expressing tumors than tumors not expressing this proinflammatory cytokine. Rather, therapeutic failure was a result of the presence of PNT. Clinical relevance for these data was suggested by the observation that myeloid cells were the predominant source of PNT in human lung, pancreatic, and breast cancer samples. Our data therefore suggest what we believe to be a novel mechanism of MDSC-mediated tumor cell resistance to CTLs.


Frequencies of regulatory T cells in the peripheral blood of dogs with primary immune-mediated thrombocytopenia and chronic enteropathy: a pilot study.
Regulatory T (Treg) cells are specialized immune cells with a pivotal role in the maintenance of immune homeostasis and control of inflammation. However, relatively little is known about immune regulation in the peripheral blood of dogs with primary immune-mediated thrombocytopenia or chronic enteropathy. Using flow cytometry this study investigated Treg responses in the peripheral blood of dogs with respective autoimmune or chronic intestinal diseases and demonstrated that a reduction of Treg frequencies is observed in dogs with clinical signs compared to dogs undergoing remission and healthy dogs. These findings suggest that reduced frequencies of Treg cells in peripheral blood might be causally associated with the onset and/or progression of autoimmune and chronic intestinal diseases in dogs and that measurement of regulatory T cells might represent a useful tool in the monitoring of treatment response and disease progression.


Genome-wide signatures of male-mediated migration shaping the Indian gene pool.
Multiple questions relating to contributions of cultural and demographical factors in the process of human geographical dispersal remain largely unanswered. India, a land of early human settlement and the resulting diversity is a good place to look for some of the answers. In this study, we explored the genetic structure of India using a diverse panel of 78 males genotyped using the GenoChip. Their genome-wide single-nucleotide polymorphism (SNP) diversity was examined in the context of various covariates that influence Indian gene pool. Admixture analysis of genome-wide SNP data showed high proportion of the Southwest Asian component in all of the Indian samples. Hierarchical clustering based on admixture proportions revealed seven distinct clusters correlating to geographical and linguistic affiliations. Convex hull overlay of Y-chromosomal haplogroups on the genome-wide SNP principal component analysis brought out distinct non-overlapping polygons of F*-M89, H*-M69, L1-M27, O2a-M95 and O3a3c1-M117, suggesting a male-mediated migration and expansion of the Indian gene pool. Lack of similar correlation with mitochondrial DNA clades indicated a shared genetic ancestry of females. We suggest that ancient male-mediated migratory events and settlement in various regional niches led to the present day scenario and peopling of India.


Prognosis and management in the first few days after a transient ischemic attack or minor ischaemic stroke.
The risk of recurrent stroke during the first few days after a transient ischaemic attack (TIA) or minor stroke is very much higher than previously estimated. However, there is considerable international variation in how patients with suspected TIA or minor stroke are managed in the acute phase, some healthcare systems providing immediate emergency inpatient care and others providing non-emergency outpatient clinic assessment. This review considers what is known about the early prognosis after TIA and minor ischaemic stroke, what factors identify individuals at particularly high early risk of stroke, and what evidence there is that urgent preventive treatment is likely to be effective in reducing the early risk of stroke.


N-methyl-D-aspartate receptor type 2B is epigenetically inactivated and exhibits tumor-suppressive activity in human esophageal cancer.
Promoter hypermethylation accompanied by gene silencing is a common feature of human cancers. We identified previously several new tumor suppressor genes based on pharmacologic unmasking of the promoter region and detection of reexpression on microarray analysis. In this study, we modified the selection of candidates from our previous microarray data by excluding genes that showed basal expression in cancer cell lines. With the new method, we found novel methylated genes with 90% accuracy. Among these 33 novel methylated genes that we identified in esophageal squamous cell carcinoma (ESCC) cell lines, N-methyl-D-aspartate receptor type 2B (NMDAR2B) was of particular interest. NMDAR2B was methylated in 95% of primary human ESCC tissue specimens and 12 ESCC cell lines by sequence analysis. NMDAR2B expression was silenced in all 12 ESCC cell lines and was reactivated by the demethylating agent 5-aza-2'-deoxycytidine. Moreover, reintroduction of the gene was accompanied by marked Ca(2+)-independent apoptosis in ESCC cell lines, suggesting that NMDAR2B can suppress tumor growth. Thus, NMDAR2B promoter methylation is common in ESCC, abrogating gene transcription and leading to cellular resistance to apoptosis.


Fish and aquatic habitat conservation in South America: a continental overview with emphasis on neotropical systems.
Fish conservation in South America is a pressing issue. The biodiversity of fishes, just as with all other groups of plants and animals, is far from fully known. Continuing habitat loss may result in biodiversity losses before full species diversity is known. In this review, the main river basins of South America (Magdalena, Orinoco, Amazon and Paraná-La Plata system), together with key aquatic habitats (mangrove-fringed estuaries of the tropical humid, tropical semi-arid and subtropical regions) are analysed in terms of their characteristics and main concerns. Habitat loss was the main concern identified for all South American ecosystems. It may be caused by damming of rivers, deforestation, water pollution, mining, poor agricultural practice or inadequate management practice. Habitat loss has a direct consequence, which is a decrease in the availability of living resources, a serious social and economic issue, especially for South American nations which are all developing countries. The introduction of exotic species and overfishing were also identified as widespread across the continent and its main freshwater, coastal and marine ecosystems. Finally, suggestions are made to find ways to overcome these problems. The main suggestion is a change of paradigm and a new design for conservation actions, starting with integrated research and aiming at the co-ordinated and harmonized management of the main transboundary waters of the continent. The actions would be focused on habitat conservation and social rescue of the less well-off populations of indigenous and non-indigenous peoples. Energy and freshwater demands will also have to be rescaled in order to control habitat loss.


Neogene precipitation, vegetation, and elevation history of the Central Andean Plateau.
Andean uplift played a fundamental role in shaping South American climate and species distribution, but the relationship between the rise of the Andes, plant composition, and local climatic evolution is poorly known. We investigated the fossil record (pollen, leaves, and wood) from the Neogene of the Central Andean Plateau and documented the earliest evidence of a puna-like ecosystem in the Pliocene and a montane ecosystem without modern analogs in the Miocene. In contrast to regional climate model simulations, our climate inferences based on fossil data suggest wetter than modern precipitation conditions during the Pliocene, when the area was near modern elevations, and even wetter conditions during the Miocene, when the cordillera was around ~1700 meters above sea level. Our empirical data highlight the importance of the plant fossil record in studying past, present, and future climates and underscore the dynamic nature of high elevation ecosystems.


Relative performance of customized and universal probe sets in target enrichment: A case study in subtribe Malinae.
Custom probe design for target enrichment in phylogenetics is tedious and often hinders broader phylogenetic synthesis. The universal angiosperm probe set Angiosperms353 may be the solution. Here, we test the relative performance of Angiosperms353 on the Rosaceae subtribe Malinae in comparison with custom probes that we specifically designed for this clade. We then address the impact of bioinformatically altering the performance of Angiosperms353 by replacing the original probe sequences with orthologs extracted from the Malus domestica genome. To evaluate the relative performance of these probe sets, we compared the enrichment efficiency, locus recovery, alignment length, proportion of parsimony-informative sites, proportion of potential paralogs, the topology and support of the resulting species trees, and the gene tree discordance. Locus recovery was highest for our custom Malinae probe set, and replacing the original Angiosperms353 sequences with a Malus representative improved the locus recovery relative to Angiosperms353. The proportion of parsimony-informative sites was similar between all probe sets, while the gene tree discordance was lower in the case of the custom probes. A custom probe set benefits from data completeness and can be tailored toward the specificities of the project of choice; however, Angiosperms353 was equally as phylogenetically informative as the custom probes. We therefore recommend using both a custom probe set and Angiosperms353 to facilitate large-scale systematic studies, where financially possible.


How many surface atoms in Co3O4 take part in oxygen evolution? Isotope labeling together with differential electrochemical mass spectrometry.
Understanding the mechanism underlying the oxygen evolution reaction (OER) on oxides is crucial for the development of many energy storage systems. Here, the mechanism of OER on a Co3O4 spinel catalyst is investigated in alkaline media using 18O-labeling combined with differential electrochemical mass spectrometry (DEMS). This work unravels the role of surface oxygen of the oxide in the OER. It is shown that in H218O-containing electrolyte the amount of 18O16O evolved increases from cycle to cycle together with a concomitant decrease of the amount of 16O2 with each cycle before reaching a steady-state value. 18O16O is also evolved from a H216O solution on a Co3O4 electrode pre-treated in H218O-containing solution, indicating the formation of the 18O-labeled oxide in the previous step. Therefore, the oxide layer takes part in OER via an oxygen exchange mechanism. The total number of oxygen atoms of the oxide participating in OER is 0.1 to 0.2% of the total oxide loading, corresponding to about 10-30% of the surface atoms; these represent the catalytically active sites. Moreover, the real surface area of the catalyst is estimated using different methods (namely the ball model, double layer capacitance method, redox peak method, isotope exchange), and compared to the BET data. The surface areas calculated from the BET data, ball model and redox peak method are similar for small particles, which indicates their smooth surface; however they are smaller than that estimated from double-layer capacitance. For larger particles, the much larger surface area estimated from the redox peak in comparison to that expected from the ball model seems to be due to their roughness. Thus, this work highlights the importance of probing the mechanism when investigating the OER activity of a catalyst.


GeneMANIA Cytoscape plugin: fast gene function predictions on the desktop.
The GeneMANIA Cytoscape plugin brings fast gene function prediction capabilities to the desktop. GeneMANIA identifies the most related genes to a query gene set using a guilt-by-association approach. The plugin uses over 800 networks from six organisms and each related gene is traceable to the source network used to make the prediction. Users may add their own interaction networks and expression profile data to complement or override the default data. The GeneMANIA Cytoscape plugin is implemented in Java and is freely available at http://www.genemania.org/plugin/.


Neurogenic orthostatic hypotension: roles of norepinephrine deficiency in its causes, its treatment, and future research directions.
Although a diversity of neurotransmitters and hormones participate in controlling blood pressure, norepinephrine released from postganglionic sympathetic nerve terminals is an important mediator of the rapid regulation of cardiovascular function required for homeostasis of cerebral perfusion. Hence, neurogenic orthostatic hypotension (NOH) often represents a deficiency of noradrenergic responsiveness to postural change. PubMed searches with 'orthostatic hypotension' and 'norepinephrine' as conjoint search terms and no restriction on language or date, so as to survey the pathophysiologic and clinical relevance of norepinephrine deficiency for current NOH interventions and for future directions in treatment and research. Norepinephrine deficiency in NOH can arise peripherally, due to cardiovascular sympathetic denervation (as in pure autonomic failure, Parkinson's disease, and a variety of neuropathies), or centrally, due to a failure of viscerosensory signals to generate adequate sympathetic traffic to intact sympathetic nerve endings (as in multiple system atrophy). Nonpharmacologic countermeasures such as pre-emptive water intake may yield blood-pressure increases exceeding those achieved pharmacologically. For patients with symptomatic NOH unresponsive to such strategies, a variety of pharmacologic interventions have been administered off-label on the basis of drug mechanisms expected to increase blood pressure via blood-volume expansion or vasoconstriction. Two pressor agents have received FDA approval: the sympathomimetic midodrine and more recently the norepinephrine prodrug droxidopa. Pressor agents are important for treating symptomatic NOH in patients unresponsive to lifestyle changes alone. However, the dysautonomia underlying NOH often permits blood-pressure excursions toward both hypotension and hypertension. Future research should aim to shed light on the resulting management issues, and should also explore the possibility of pharmacotherapy selectively targeting orthostatic blood-pressure decreases.


Short dysfunctional telomeres impair tumorigenesis in the INK4a(delta2/3) cancer-prone mouse.
Maintenance of telomere length is predicted to be essential for bypass of senescence and crisis checkpoints in cancer cells. The impact of telomere dysfunction on tumorigenesis was assessed in successive generations of mice doubly null for the telomerase RNA (mTR) and the INK4a tumor suppressor genes. Significant reductions in tumor formation in vivo and oncogenic potential in vitro were observed in late generations of telomerase deficiency, coincident with severe telomere shortening and associated dysfunction. Reintroduction of mTR into cells significantly restored the oncogenic potential, indicating telomerase activation is a cooperating event in the malignant transformation of cells containing critically short telomeres. The results described here demonstrate that loss of telomere function in a cancer-prone mouse model possessing intact DNA damage responses impairs, but does not prevent, tumor formation.


'I fell off and landed badly': Children's experiences of forearm fracture and injury prevention.
Forearm fractures are one of the most common injuries sustained by children. Our descriptive study addressed, from the perspective of a child, the following research objectives: (1) to describe their fracture experience and (2) to describe how fractures might be prevented. Photovoice is a unique research strategy by which people create and discuss photographs. This technique has been used to elicit the perspectives of those whose voices are often 'not heard' in research, like children. Participants were recruited from a larger three-year prospective trial and included 10 boys (12.3 ± 1.6 years) and 7 girls (11.3 ± 1.6 years). We asked participants to take pictures to explain where their injury occurred (place), what they were doing at the time (context) and how the fracture had happened (mechanism). We also used semi-structured interview techniques. The following key themes emerged from our interviews: (1) the built environment as a key factor that 'caused' their fracture, (2) the fracture experienced as a journey not an event and (3) strategies to prevent fractures. A simple clinical step to potentially reduce subsequent fractures will be for clinicians to have a brief conversation with their young patients and to listen to the child's personal preventive strategies.


Foam flow in a model porous medium: I. The effect of foam coarsening.
Foam structure evolves with time due to gas diffusion between bubbles (coarsening). In a bulk foam, coarsening behaviour is well defined, but there is less understanding of coarsening in confined geometries such as porous media. Previous predictions suggest that coarsening will cause foam lamellae to move to low energy configurations in the pore throats, resulting in greater capillary resistance when restarting flow. Foam coarsening experiments were conducted in both a model-porous-media micromodel and in a sandstone core. In both cases, foam was generated by coinjecting surfactant solution and nitrogen. Once steady state flow had been achieved, the injection was stopped and the system sealed off. In the micromodel, the foam coarsening was recorded using time-lapse photography. In the core flood, the additional driving pressure required to reinitiate flow after coarsening was measured. In the micromodel the bubbles coarsened rapidly to the pore size. At the completion of coarsening the lamellae were located in minimum energy configurations in the pore throats. The wall effect meant that the coarsening did not conform to the unconstricted growth laws. The coreflood tests also showed coarsening to be a rapid process. The additional driving pressure to restart flow reached a maximum after just 2 minutes.


The effects of food and parasitism on reproductive performance of a wild rodent.
Food and parasitism can have complex effects on small mammal reproduction. In this study, we tested the effects of sex, food, and parasitism on reproductive performance of the Taiwan field mouse (Apodemus semotus). In a field experiment, we increased food availability for a portion of the mice in the population by providing sorghum seeds to a set of food stations. We reduced parasite intensity of randomly chosen mice through ivermectin treatment. We determined the number and quality of offspring for the mice using paternity analysis. We quantified seed consumption with stable carbon isotope values of mouse plasma and parasite intensity with fecal egg counts of intestinal nematodes and cestodes (FEC). In a laboratory experiment, we reduced parasite intensity of randomly chosen mice through ivermectin treatment. We quantified their immune functions by total white blood cell count, percent granulocyte count, and percent lymphocyte count through hematological analyses. We measured the FEC and energy intake of the mice. From the field experiment, the number of offspring in A. semotus increased with increasing seed consumption. Due to the trade-off between number and quality of offspring, the offspring quality decreased with increasing seed consumption for the females. The ivermectin treatment did not affect offspring number or quality. However, the FEC was positively correlated with number of offspring. In the laboratory experiment, the percent lymphocyte/granulocyte count changed with parasite intensity at low energy intake, which was relaxed at high energy intake. This study demonstrated positive effects of food availability and neutral effects of parasitism on A. semotus reproduction. However, the benefits of food availability for the females need to take into account the offspring number-quality trade-off, and at high infection intensity, parasitism might negatively affect offspring quality for the males. We suggest that food availability could mediate the relationships between parasite intensity and immune responses.


Nitric oxide as a biomarker of intracellular Salmonella viability and identification of the bacteriostatic activity of protein kinase A inhibitor H-89.
Salmonella enterica serovar Enteritidis is one of the most prevalent Salmonella serovars in poultry and is often associated with human salmonellosis. S. Enteritidis is known to suppress nitric oxide (NO) production in infected chicken macrophage HD11 cells, while dead S. Enteritidis stimulates a high level of NO production, suggesting a bacterial inhibitory effect on NO production. Based on these observations, the present study was conducted to evaluate whether NO production in S. Enteritidis-infected HD11 cells can be used as a biomarker to identify molecules that kill intracellular Salmonella. Since Salmonella are known to manipulate the host cell kinase network to facilitate intracellular survival, we screened a group of pharmaceutical inhibitors of various kinases to test our hypothesis. A protein kinase A inhibitor, H-89, was found to reverse the suppression of NO production in S. Enteritidis-infected HD11 cells. Production of NO in S. Enteritidis-infected HD11 cells increased significantly following treatment with H-89 at or above 20 µM. Inversely, the number of viable intracellular Salmonella decreased significantly in cells treated with H-89 at or above 30 µM. Furthermore, the growth rate of S. Enteritidis in culture was significantly inhibited by H-89 at concentrations from 20 to 100 µM. Our results demonstrate that NO-based screening using S. Enteritidis-infected HD11 cells is a viable tool to identify chemicals with anti-intracellular Salmonella activity. Using this method, we have shown H-89 has bacteriostatic activity against Salmonella, independent of host cell protein kinase A or Akt1 activity.


Geropsychiatric education programs. Providing skills and understanding.
1. The treatment of mental health problems is a growing concern in long-term care facilities; up to 80% of nursing home residents suffer from some type of mental illness, but most receive no active treatment. 2. Nursing home staff do not receive sufficient training on mental health issues, the aging process, and assessment and management of psychiatric symptoms in the elderly. 3. Psychiatric nurses can provide a number of quality geriatric mental health programs to increase staff knowledge and promote quality of care.


Using simulation models to evaluate ape nest survey techniques.
Conservationists frequently use nest count surveys to estimate great ape population densities, yet the accuracy and precision of the resulting estimates are difficult to assess. We used mathematical simulations to model nest building behavior in an orangutan population to compare the quality of the population size estimates produced by two of the commonly used nest count methods, the 'marked recount method' and the 'matrix method.' We found that when observers missed even small proportions of nests in the first survey, the marked recount method produced large overestimates of the population size. Regardless of observer reliability, the matrix method produced substantial overestimates of the population size when surveying effort was low. With high observer reliability, both methods required surveying approximately 0.26% of the study area (0.26 km(2) out of 100 km(2) in this simulation) to achieve an accurate estimate of population size; at or above this sampling effort both methods produced estimates within 33% of the true population size 50% of the time. Both methods showed diminishing returns at survey efforts above 0.26% of the study area. The use of published nest decay estimates derived from other sites resulted in widely varying population size estimates that spanned nearly an entire order of magnitude. The marked recount method proved much better at detecting population declines, detecting 5% declines nearly 80% of the time even in the first year of decline. These results highlight the fact that neither nest surveying method produces highly reliable population size estimates with any reasonable surveying effort, though either method could be used to obtain a gross population size estimate in an area. Conservation managers should determine if the quality of these estimates are worth the money and effort required to produce them, and should generally limit surveying effort to 0.26% of the study area, unless specific management goals require more intensive sampling. Using site- and time- specific nest decay rates (or the marked recount method) are essential for accurate population size estimation. Marked recount survey methods with sufficient sampling effort hold promise for detecting population declines.


White matter microstructure across the adult lifespan: A mixed longitudinal and cross-sectional study using advanced diffusion models and brain-age prediction.
The macro- and microstructural architecture of human brain white matter undergoes substantial alterations throughout development and ageing. Most of our understanding of the spatial and temporal characteristics of these lifespan adaptations come from magnetic resonance imaging (MRI), including diffusion MRI (dMRI), which enables visualisation and quantification of brain white matter with unprecedented sensitivity and detail. However, with some notable exceptions, previous studies have relied on cross-sectional designs, limited age ranges, and diffusion tensor imaging (DTI) based on conventional single-shell dMRI. In this mixed cross-sectional and longitudinal study (mean interval: 15.2 months) including 702 multi-shell dMRI datasets, we combined complementary dMRI models to investigate age trajectories in healthy individuals aged 18 to 94 years (57.12% women). Using linear mixed effect models and machine learning based brain age prediction, we assessed the age-dependence of diffusion metrics, and compared the age prediction accuracy of six different diffusion models, including diffusion tensor (DTI) and kurtosis imaging (DKI), neurite orientation dispersion and density imaging (NODDI), restriction spectrum imaging (RSI), spherical mean technique multi-compartment (SMT-mc), and white matter tract integrity (WMTI). The results showed that the age slopes for conventional DTI metrics (fractional anisotropy [FA], mean diffusivity [MD], axial diffusivity [AD], radial diffusivity [RD]) were largely consistent with previous research, and that the highest performing advanced dMRI models showed comparable age prediction accuracy to conventional DTI. Linear mixed effects models and Wilk's theorem analysis showed that the 'FA fine' metric of the RSI model and 'orientation dispersion' (OD) metric of the NODDI model showed the highest sensitivity to age. The results indicate that advanced diffusion models (DKI, NODDI, RSI, SMT mc, WMTI) provide sensitive measures of age-related microstructural changes of white matter in the brain that complement and extend the contribution of conventional DTI.


Antiretroviral Drug Use in a Cohort of HIV-Uninfected Women in the United States: HIV Prevention Trials Network 064.
Antiretroviral (ARV) drug use was analyzed in HIV-uninfected women in an observational cohort study conducted in 10 urban and periurban communities in the United States with high rates of poverty and HIV infection. Plasma samples collected in 2009-2010 were tested for the presence of 16 ARV drugs. ARV drugs were detected in samples from 39 (2%) of 1,806 participants: 27/181 (15%) in Baltimore, MD and 12/179 (7%) in Bronx, NY. The ARV drugs detected included different combinations of non-nucleoside reverse transcriptase inhibitors and protease inhibitors (1-4 drugs/sample). These data were analyzed in the context of self-reported data on ARV drug use. None of the 39 women who had ARV drugs detected reported ARV drug use at any study visit. Further research is needed to evaluate ARV drug use by HIV-uninfected individuals.


Outcomes in the highest panel reactive antibody recipients of deceased donor kidneys under the new kidney allocation system.
Since the institution of the new kidney allocation system in December 2014, kidney transplant candidates with the highest calculated panel reactive antibodies (cPRA) of 99-100 have been transplanted at much higher rates. However, concerns have been raised that outcomes in these patients might be impaired due to higher immunological risk and longer cold ischemia times resulting from long-distance sharing of kidneys. Here, we compare outcomes at the University of Wisconsin between study patients with cPRA 99-100 and all other recipients of deceased donor kidneys transplanted between 12/04/2014 and 12/31/2015. All patients had at least 6 months post-transplant follow-up. The mean follow-up was 13.9±3 months in cPRA ≥99% and 12.3±3.5 months in cPRA ≤98%. There was a total of 152 transplants, 25 study patients, and 127 controls. No statistically significant differences were found between the two groups in delayed graft function, rejection, kidney function, graft and patient survival, or infections. We conclude that transplanting the most highly sensitized patients with kidneys shared outside their local donation service areas is associated with excellent short-term outcomes that are comparable to controls.


Evaluation of a self-adhering flowable composite in terms of micro-shear bond strength and microleakage.
The objective of this study was to evaluate dentin bond strength and microleakage of a self-adhering flowable resin with or without adhesive resin. Dentin pieces were randomly assigned to three groups (n = 10): Group OR (Optibond+Revolution Formula 2 Flow); Group VF (Vertise Flow); and Group OVF (Optibond+Vertise Flow). Then specimens were stressed in shear at a cross-head speed of 1 mm/min until the bond failed. Moreover, microleakage was determined by restoring Class V cavities. A statistical analyses was carried out using Mann-Whitney U and Kruskal-Wallis tests (p < 0.05). Group OVF specimens produced the highest mean shear bond strength. In addition, the results showed that the marginal seal in group VF was similar to that in group OR. Self-adhering flowable composite resin combined with adhesive resin provided stronger dentin bond strength and a better marginal seal than when it was used individually. VF is a useful material with an acceptable bond strength and marginal seal.


Basic Characteristics of Adults with Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Adenopathy Syndrome in Comparison with the Typical Pediatric Expression of Disease.
Autoinflammatory diseases are caused by inflammasome dysregulation leading to overproduction of proinflammatory cytokines and a pathological delay in the inflammation switching off. The progress of cellular biology has partially clarified pathogenic mechanisms behind monogenic autoinflammatory diseases, whereas little is known about the polygenic ones. Although the genetic susceptibility of periodic fever, aphthous stomatitis, pharyngitis, and adenopathy (PFAPA) syndrome is still obscure, the presence of overlapping symptoms with monogenic periodic fevers, the recurrence in family members, the important role played by dysregulated interleukin- (IL-) 1β secretion during flares, the overexpression of inflammasome-associated genes during attacks, and, last but not least, the therapeutic efficacy of IL-1β blockade strongly indicate a potential genetic involvement in its pathogenesis, probably linked with environmental factors. PFAPA syndrome has a typical inception in the pediatric age, but a delayed onset during adulthood has been described as well. Treatments required as well as effectiveness of tonsillectomy remain controversial, even if the disease seems to have a self-limited course mostly in children. The purpose of this review is to provide an overview of this complex polygenic/multifactorial autoinflammatory disorder in which the innate immune system undoubtedly plays a basic role.


Interleukin-1beta (IL-1beta) processing pathway.
The innate immune system senses molecular patterns from invading microorganisms. Once activated, it orchestrates the inflammatory response by secreting proinflammatory cytokines, such as interleukin-1 (IL-1)-type cytokines, in particular IL-1beta. IL-1 mediates the expression of a vast array of genes involved in secondary inflammation. IL-1-responsive genes coordinate all aspects of local inflammation and also attract and activate cells of the adaptive immune system at sites of infection. Moreover, the innate immune system can also sense a wide range of nonmicrobial molecular patterns that represent danger or damage signals. These signals activate the NALP3-inflammasome pathway, which plays a central role in acute and chronic sterile inflammation. Here, we describe the essential components of the NALP3-inflammasome that control processing and release of IL-1beta.


Superposition of transcriptional behaviors determines gene state.
We introduce a novel technique to determine the expression state of a gene from quantitative information measuring its expression. Adopting a productive abstraction from current thinking in molecular biology, we consider two expression states for a gene--Up or Down. We determine this state by using a statistical model that assumes the data behaves as a combination of two biological distributions. Given a cohort of hybridizations, our algorithm predicts, for the single reading, the probability of each gene's being in an Up or a Down state in each hybridization. Using a series of publicly available gene expression data sets, we demonstrate that our algorithm outperforms the prevalent algorithm. We also show that our algorithm can be used in conjunction with expression adjustment techniques to produce a more biologically sound gene-state call. The technique we present here enables a routine update, where the continuously evolving expression level adjustments feed into gene-state calculations. The technique can be applied in almost any multi-sample gene expression experiment, and holds equal promise for protein abundance experiments.


Monomethyl Fumarate Protects the Retina From Light-Induced Retinopathy.
We determine if monomethyl fumarate (MMF) can protect the retina in mice subjected to light-induced retinopathy (LIR). Albino BALB/c mice were intraperitoneally injected with 50 to 100 mg/kg MMF before or after exposure to bright white light (10,000 lux) for 1 hour. Seven days after light exposure, retinal structure and function were evaluated by optical coherence tomography (OCT) and electroretinography (ERG), respectively. Retinal histology also was performed to evaluate photoreceptor loss. Expression levels of Hcar2 and markers of microglia activation were measured by quantitative PCR (qPCR) in the neural retina with and without microglia depletion. At 24 hours after light exposure, retinal sections and whole mount retinas were stained with Iba1 to evaluate microglia status. The effect of MMF on the nuclear factor kB subunit 1 (NF-kB) and Nrf2 pathways was measured by qPCR and Western blot. MMF administered before light exposure mediated dose-dependent neuroprotection in a mouse model of LIR. A single dose of 100 mg/kg MMF fully protected retinal structure and function without side effects. Expression of the Hcar2 receptor and the microglia marker Cd14 were upregulated by LIR, but suppressed by MMF. Depleting microglia reduced Hcar2 expression and its upregulation by LIR. Microglial activation, upregulation of proinflammatory genes (Nlrp3, Caspase1, Il-1β, Tnf-α), and upregulation of antioxidative stress genes (Hmox1) associated with LIR were mitigated by MMF treatment. MMF can completely protect the retina from LIR in BALB/c mice. Expression of Hcar2, the receptor of MMF, is microglia-dependent in the neural retina. MMF-mediated neuroprotection was associated with attenuation of microglia activation, inflammation and oxidative stress in the retina.


Auditory-visual integration of emotional signals in a virtual environment for cynophobia.
Cynophobia (dog phobia) has both visual and auditory relevant components. In order to investigate the efficacy of virtual reality (VR) exposure-based treatment for cynophobia, we studied the efficiency of auditory-visual environments in generating presence and emotion. We conducted an evaluation test with healthy participants sensitive to cynophobia in order to assess the capacity of auditory-visual virtual environments (VE) to generate fear reactions. Our application involves both high fidelity visual stimulation displayed in an immersive space and 3D sound. This specificity enables us to present and spatially manipulate fearful stimuli in the auditory modality, the visual modality and both. Our specific presentation of animated dog stimuli creates an environment that is highly arousing, suggesting that VR is a promising tool for cynophobia treatment and that manipulating auditory-visual integration might provide a way to modulate affect.


Exome sequencing in 53 sporadic cases of schizophrenia identifies 18 putative candidate genes.
Schizophrenia (SCZ) is a severe, debilitating mental illness which has a significant genetic component. The identification of genetic factors related to SCZ has been challenging and these factors remain largely unknown. To evaluate the contribution of de novo variants (DNVs) to SCZ, we sequenced the exomes of 53 individuals with sporadic SCZ and of their non-affected parents. We identified 49 DNVs, 18 of which were predicted to alter gene function, including 13 damaging missense mutations, 2 conserved splice site mutations, 2 nonsense mutations, and 1 frameshift deletion. The average number of exonic DNV per proband was 0.88, which corresponds to an exonic point mutation rate of 1.7×10(-8) per nucleotide per generation. The non-synonymous-to-synonymous mutation ratio of 2.06 did not differ from neutral expectations. Overall, this study provides a list of 18 putative candidate genes for sporadic SCZ, and when combined with the results of similar reports, identifies a second proband carrying a non-synonymous DNV in the RGS12 gene.


Comparison of plasma vitamin A and E, copper and zinc levels in free-ranging and captive greater flamingos (Phoenicopterus roseus) and their relation to pododermatitis.
Pododermatitis is a worldwide problem in captive flamingos. Studies in domestic poultry showed that nutrition is a possible influencing factor for pododermatitis. Vitamin A and E, copper and zinc levels were analysed in two different diets (diet 1 = in-house mix and diet 2 = commercial diet) and in plasma of captive greater flamingos fed these diets and compared to those of free-ranging greater flamingos. Results were analysed with respect to type and severity of foot lesions of the individuals from the different groups. Juvenile and subadult/adult captive flamingos on diet 1 showed various types and severities of foot lesions, whereas no foot lesions were found at the time of blood sampling in juvenile captive flamingos on diet 2. Juvenile captive flamingos on diet 1 had significantly lower plasma zinc levels than juvenile captive flamingos on diet 2 and juvenile free-ranging flamingos; data were also lower than reference ranges for flamingos, poultry and cranes. There were no significant differences in plasma vitamin A, vitamin E, copper or zinc levels between animals with different types of foot lesions or with different severity scores. Shortly after the change to diet 2 (fed to juvenile captive flamingos that did not show any foot lesion), the flooring of the outdoor water pools was covered with fine granular sand. Because both factors (nutrition and flooring) were changed during the same evaluation period, it cannot be concluded which factor contributed in what extent to the reduction of foot lesions. While it is assumed that low plasma zinc levels identified in the group of juvenile captive flamingos on diet 1 were not directly responsible for foot lesions observed in these animals, they may have played a role in altering the skin integrity of the feet and predisposing them to pododermatitis.


Does specialist physician supply affect pediatric asthma health outcomes?
Pediatrician and pediatric subspecialist density varies substantially among the various Canadian provinces, as well as among various states in the US. It is unknown whether this variability impacts health outcomes. To study this knowledge gap, we evaluated pediatric asthma admission rates within the 2 Canadian provinces of Manitoba and Saskatchewan, which have similarly sized pediatric populations and substantially different physician densities. This was a retrospective cross-sectional cohort study. Health regions defined by the provincial governments, have, in turn, been classified into "peer groups" by Statistics Canada, on the basis of common socio-economic characteristics and socio-demographic determinants of health. To study the relationship between the distribution of the pediatric workforce and health outcomes in Canadian children, asthma admission rates within comparable peer group regions in both provinces were examined by combining multiple national and provincial health databases. We generated physician density maps for general practitioners, and general pediatricians practicing in Manitoba and Saskatchewan in 2011. At the provincial level, Manitoba had 48.6 pediatricians/100,000 child population, compared to 23.5/100,000 in Saskatchewan. There were 3.1 pediatric asthma specialists/100,000 child population in Manitoba and 1.4/100,000 in Saskatchewan. Among peer-group A, the differences were even more striking. A significantly higher number of patients were admitted in Saskatchewan (590.3/100,000 children) compared to Manitoba (309.3/100,000, p < 0.0001). Saskatchewan, which has a lower pediatrician and pediatric asthma specialist supply, had a higher asthma admission rate than Manitoba. Our data suggest that there is an inverse relationship between asthma admissions and pediatrician and asthma specialist supply.


NK-B cell cross talk induces CXCR5 expression on natural killer cells.
B cell follicles (BCFs) in lymph nodes (LNs) are generally exempt of CD8+ T and NK cells. African green monkeys (AGMs), a natural host of simian immunodeficiency virus (SIV), display NK cell-mediated viral control in BCF. NK cell migration into BCF in chronically SIVagm-infected AGM is associated with CXCR5+ NK cells. We aimed to identify the mechanism leading to CXCR5 expression on NK cells. We show that CXCR5+ NK cells in LN were induced following SIVagm infection. CXCR5+ NK cells accumulated preferentially in BCF with proliferating B cells. Autologous NK-B cell co-cultures in transwell chambers induced CXCR5+ NK cells. Transcriptome analysis of CXCR5+ NK cells revealed expression of bcl6 and IL6R. IL-6 induced CXCR5 on AGM and human NK cells. IL6 mRNA was detected in LN at higher levels during SIVagm than SIVmac infection and often produced by plasma cells. Our study reveals a mechanism of B cell-dependent NK cell regulation.


Face scanning and spontaneous emotion preference in Cornelia de Lange syndrome and Rubinstein-Taybi syndrome.
Existing literature suggests differences in face scanning in individuals with different socio-behavioural characteristics. Cornelia de Lange syndrome (CdLS) and Rubinstein-Taybi syndrome (RTS) are two genetically defined neurodevelopmental disorders with unique profiles of social behaviour. Here, we examine eye gaze to the eye and mouth regions of neutrally expressive faces, as well as the spontaneous visual preference for happy and disgusted facial expressions compared to neutral faces, in individuals with CdLS versus RTS. Results indicate that the amount of time spent looking at the eye and mouth regions of faces was similar in 15 individuals with CdLS and 17 individuals with RTS. Both participant groups also showed a similar pattern of spontaneous visual preference for emotions. These results provide insight into two rare, genetically defined neurodevelopmental disorders that have been reported to exhibit contrasting socio-behavioural characteristics and suggest that differences in social behaviour may not be sufficient to predict attention to the eye region of faces. These results also suggest that differences in the social behaviours of these two groups may be cognitively mediated rather than subcortically mediated.


Phenotypic and genotypic resistance patterns of HIV-1 isolates derived from individuals treated with didanosine and stavudine.
To evaluate the phenotypic susceptibilities and genotypic resistance patterns to both didanosine and stavudine of baseline and follow-up HIV-1 isolate pairs, derived from antiretroviral naive subjects treated with this dual nucleoside combination. Phenotypic drug susceptibility testing was performed in peripheral blood mononuclear cells on 34 viral isolate pairs derived from patients participating in the BMS AI-460 trial. Sequencing of the complete reverse transcriptase of 36 study isolate pairs, baseline and follow-up, was performed using standard dideoxy techniques. The mean fold change in susceptibilities to didanosine was 1.6 (P= 0.278) and to stavudine 1.9 (P= 0.002, Wilcoxon's signed rank test). Mutations classically associated with zidovudine resistance were observed to emerge in 7 out of 36 isolates, T215Y/F (four), M41L +T215Y/F (two) and D67N (one). Other mutations observed included the A62V, V751, F77L, F116Y, Q151 M multinucleoside resistance complex (one), the Q151M mutation (two) and the rare V75T mutation (two). No mutations classically associated with didanosine exposure and resistance were observed. No relationship was evident between the emergence of zidovudine associated mutations and the level of phenotypic resistance to either stavudine or didanosine or between the emergence of zidovudine associated mutations and changes in plasma HIV RNA levels. These comprehensive data demonstrate modest (< twofold) mean reductions in didanosine and stavudine susceptibilities at follow-up. The emergence of zidovudine associated mutations in this retroviral-naive population treated with combination didanosine and stavudine therapy is notable. Furthermore, the emergence of these mutations and of the Q151 M multinucleoside resistance complex raise concerns for potential nucleoside analog cross-resistance. The potential mechanisms driving the selection of the zidovudine associated mutations in the setting of didanosine and stavudine therapy and the relevance of these findings to current three and four drug regimens merit further evaluation.


An improved physical and genetic map of the genome of alkaliphilic Bacillus sp. C-125.
Among alkaliphilic bacteria reported so far, Bacillus sp. C-125 is the strain most thoroughly characterized physiologically, biochemically, and genetically. A physical map of the chromosome of this strain was constructed to facilitate further genome analysis, and the genome size was revised from 3.7 to 4.25Mb. Complete digestion of the chromosomal DNA with two rare cut restriction endonucleases, AscI and Sse8387I, each yielded 20 fragments ranging in size from 20 to 600 kb. Seventeen linking clones were isolated in each instance to join the adjacent AscI or Sse8387I fragments in the chromosomal map. All AscI linking clones isolated were sequenced and analyzed by comparison with the BSORF database to map the genes in the chromosome of strain C-125. Several ORFs showing significant similarities to those of B. subtilis in the AscI linking clones were positioned on the physical map. The oriC region of the C-125 chromosome was identified by southern blot analysis with a DNA probe containing the gyrB region.


Central nervous system infarction related to cocaine abuse.
Cocaine use in the United States has reached epidemic proportions, and increased availability of "crack" since 1983 has noticeably increased the incidence of neurovascular complications. In this report, we examine the relationship between cocaine use and ischemic infarct. This study reports 18 cases of ischemic cerebrovascular events, which occurred among 15 men and three women aged 21-47 years who were evaluated in a 2-year period. Clinical presentations include thirteen cases with hemispheric infarcts, two brain stem strokes, two anterior spinal artery infarcts, and one with both hemispheric and cerebellar infarcts. Nine patients smoked crack, four snorted cocaine, and three injected it intravenously. In two cases, the route of administration could not be determined. Two patients died, but the other survived with various degrees of neurological deficit. Traditional risk factors for strokes were identified in only six patients, suggesting that these factors are not necessary for the occurrence of a cocaine-related infarct. Multiple overlapping mechanisms may be responsible, including vasospasm, sudden onset of hypertension, myocardial infarction with cardiac arrhythmias, increased platelet aggregation, and vasculitis.


A Be-type star with a black-hole companion.
Stellar-mass black holes have all been discovered through X-ray emission, which arises from the accretion of gas from their binary companions (this gas is either stripped from low-mass stars or supplied as winds from massive ones). Binary evolution models also predict the existence of black holes accreting from the equatorial envelope of rapidly spinning Be-type stars (stars of the Be type are hot blue irregular variables showing characteristic spectral emission lines of hydrogen). Of the approximately 80 Be X-ray binaries known in the Galaxy, however, only pulsating neutron stars have been found as companions. A black hole was formally allowed as a solution for the companion to the Be star MWC 656 (ref. 5; also known as HD 215227), although that conclusion was based on a single radial velocity curve of the Be star, a mistaken spectral classification and rough estimates of the inclination angle. Here we report observations of an accretion disk line mirroring the orbit of MWC 656. This, together with an improved radial velocity curve of the Be star through fitting sharp Fe II profiles from the equatorial disk, and a refined Be classification (to that of a B1.5-B2 III star), indicates that a black hole of 3.8 to 6.9 solar masses orbits MWC 656, the candidate counterpart of the γ-ray source AGL J2241+4454 (refs 5, 6). The black hole is X-ray quiescent and fed by a radiatively inefficient accretion flow giving a luminosity less than 1.6 × 10(-7) times the Eddington luminosity. This implies that Be binaries with black-hole companions are difficult to detect in conventional X-ray surveys.


Complete genome of biodegradable plastics-decomposing Roseateles depolymerans KCTC 42856(T) (=61A(T)).
Roseateles depolymerans is a Gram-negative, rod-shaped, flagellated, obligately aerobic, photosynthetic bacterium that was isolated from the Hanamuro River, Ibaraki Prefecture, Japan. Here, we report the complete genome of R. depolymerans KCTC 42856(T) (=61A(T)=DSM 11813(T)=CCUG 48747(T)=NCIMB 13588(T)), which consists of 5,681,722bp (G+C content of 66.57%) with a single chromosome, 4,773 protein-coding genes, 57 tRNAs and 4 rRNA operons. Several genes related to degradation of aliphatic and aromatic polymers were detected in the genome that help explain how the strain mediates decomposition of biodegradable plastics into fragments which are then assimilated and subsequently metabolized by microbial cells.


Heterotrophic Bacteria Dominate the Diazotrophic Community in the Eastern Indian Ocean (EIO) during Pre-Southwest Monsoon.
The diazotrophic communities play an important role in sustaining primary productivity through adding new nitrogen to oligotrophic marine ecosystems. Yet, their composition in the oligotrophic Indian Ocean is poorly understood. Here, we report the first observation of phylogenetic diversity and distribution of diazotrophs in the Eastern Indian Ocean (EIO) surface water (to 200 m) during the pre-southwest monsoon period. Through high throughput sequencing of nifH genes, we identified diverse groups of diazotrophs in the EIO including both non-cyanobacterial and cyanobacterial phylotypes. Proteobacteria (mainly Alpha-, Beta-, and Gamma-proteobacteria) were the most diverse and abundant groups within all the diazotrophs, which accounted for more than 86.9% of the total sequences. Cyanobacteria were also retrieved, and they were dominated by the filamentous non-heterocystous cyanobacteria Trichodesmium spp. Other cyanobacteria such as unicellular diazotrophic cyanobacteria were detected sporadically. Interestingly, our qPCR analysis demonstrated that the depth-integrated gene abundances of the diazotrophic communities exhibited spatial heterogeneity with Trichodesmium spp. appeared to be more abundant in the Bay of Bengal (p < 0.05), while Sagittula castanea (Alphaproteobacteria) was found to be more dominating in the equatorial region and offshores (p < 0.05). Non-metric multidimensional scaling analysis (NMDS) further confirmed distinct vertical and horizontal spatial variations in the EIO. Canonical correspondence analysis (CCA) indicated that temperature, salinity, and phosphate were the major environmental factors driving the distribution of the diazotroph communities. Overall, our study provides the first insight into the diversity and distribution of the diazotrophic communities in EIO. The findings from this study highlight distinct contributions of both non-cyanobacteria and cyanobacteria to N2 fixation. Moreover, our study reveals information that is critical for understanding spatial heterogeneity and distribution of diazotrophs, and their vital roles in nitrogen and carbon cycling.


Time of Day and Day of Week Trends in EMS Demand.
We examined temporal variations in overall Emergency Medical Services (EMS) demand, as well as medical and trauma cases separately. We analyzed cases according to time of day and day of week to determine whether population level demand demonstrates temporal patterns that will increase baseline knowledge for EMS planning. We conducted a secondary analysis of data from the Ambulance Victoria data warehouse covering the period 2008-2011. We included all cases of EMS attendance which resulted in 1,203,803 cases for review. Data elements comprised age, gender, date and time of call to the EMS emergency number along with the clinical condition of the patient. We employed Poisson regression to analyze case numbers and trigonometric regression to quantify distribution patterns. EMS demand exhibited a bimodal distribution with the highest peak at 10:00 and a second smaller peak at 19:00. The highest number of cases occurred on Fridays, and the lowest on Tuesdays and Wednesdays. However, the distribution of cases throughout the day differed by day of week. Distribution patterns on Fridays, Saturdays and Sundays differed significantly from the rest of the week (p < 0.001). When categorized into medical or trauma cases, medical cases were more frequent during working hours and involved patients of higher mean age (57 years vs. 49 years for trauma, p < 0.001). Trauma cases peaked on Friday and Saturday nights around midnight. Day of week EMS demand distribution patterns reveal differences that can be masked in aggregate data. Day of week EMS demand distribution patterns showed not only which days have differences in demand but the times of day at which the demand changes. Patterns differed by case type as well. These differences in distribution are important for EMS demand planning. Increased understanding of EMS demand patterns is imperative in a climate of ever-increasing demand and fiscal constraints. Further research is needed into the effect of age and case type on EMS demand.


Cost-effectiveness of tissue-type plasminogen activator in the 3- to 4.5-hour time window for acute ischemic stroke.
The aim of this study was to determine the cost-effectiveness of tissue-type plasminogen activator (tPA) treatment in the 3- to 4.5-hour time window after ischemic stroke. Decision-analytic and Markov state-transition models were created to determine the cost-effectiveness of treatment of ischemic stroke patients with intravenous tPA administered in the 3- to 4.5-hour time window compared with medical therapy without tPA. Health benefits were measured in quality-adjusted life-years (QALYs). The economic outcome measure of the model was the difference in estimated healthcare costs between the 2 treatment alternatives. The incremental cost-effectiveness ratio was calculated by dividing the cost difference by the difference in QALYs. One-way sensitivity and probabilistic analyses were performed to test the robustness of the model. The administration of tPA compared with standard medical therapy resulted in a lifetime gain of 0.28 QALYs for an additional cost of $6050, yielding an incremental cost-effectiveness ratio of $21 978 per QALY. One-way sensitivity analyses demonstrated that the incremental cost-effectiveness ratio was most sensitive to the cost of hospitalization for patients who received tPA. Based on probabilistic analysis, there is an 88% probability that tPA is the preferred treatment at a willingness-to-pay threshold of $50 000 per QALY. The balance of costs and benefits favors treatment with intravenous tPA in the 3- to 4.5-hour time window. This supports, from a societal perspective, the use of tPA therapy in this treatment time window for acute ischemic stroke.


The eyes remember it: oculography and pupillometry during recollection in three amnesic patients.
Two patients (TC and SS) with lesions that included the hippocampal regions (predominantly on the left side) were severely impaired in their recall of simple, verbally stated facts. However, both patients remembered spatial information that was temporally associated with semantic information. Specifically, TC and SS could not recall explicitly the content of an episode, but their spontaneous oculomotor behavior showed that they retained some information about the event as their gaze automatically returned to the locations on the computer screen where visual information had been paired to verbally presented information. Thus, this spatial information is implicit, automatically retrieved, and eye-based, as when one patient (TC) was asked to point with the finger to the same positions he was impaired. In addition, in an old/new recognition task, TC and SS and an additional patient, OB, showed significant changes in eye pupil diameter when viewing novel visual stimuli compared to stimuli that they had previously seen, also when they (incorrectly) declared with confidence that an old item was new. The spared memory of these patients, despite severe amnesia for the learning episodes, is characterized by a re-enactment of previous eye fixations that were associated with each (forgotten) episode and physiological responses (as indexed by pupillometry) to previously seen stimuli. Such spared memory can be seen as a type of "snapshot" memory, which automatically processes eye-based spatial information and whose content remains implicit. Finally, we surmise on the basis of the neuroanatomical findings of these patients, that neural substrates in the spared (right) hemisphere might support both the eye fixations' re-enactment and implicit visual pattern recognition.


Benefits and risks of the Sanofi-Pasteur dengue vaccine: Modeling optimal deployment.
The first approved dengue vaccine has now been licensed in six countries. We propose that this live attenuated vaccine acts like a silent natural infection in priming or boosting host immunity. A transmission dynamic model incorporating this hypothesis fits recent clinical trial data well and predicts that vaccine effectiveness depends strongly on the age group vaccinated and local transmission intensity. Vaccination in low-transmission settings may increase the incidence of more severe "secondary-like" infection and, thus, the numbers hospitalized for dengue. In moderate transmission settings, we predict positive impacts overall but increased risks of hospitalization with dengue disease for individuals who are vaccinated when seronegative. However, in high-transmission settings, vaccination benefits both the whole population and seronegative recipients. Our analysis can help inform policy-makers evaluating this and other candidate dengue vaccines.


Guideline No. 422b: Menopause and Genitourinary Health.
Provide strategies for improving the care of perimenopausal and postmenopausal women based on the most recent published evidence. Perimenopausal and postmenopausal women. Target population will benefit from the most recent published scientific evidence provided via the information from their health care provider. No harms or costs are involved with this information since women will have the opportunity to choose among the different therapeutic options for the management of the symptoms and morbidities associated with menopause, including the option to choose no treatment. Databases consulted were PubMed, MEDLINE, and the Cochrane Library for the years 2002-2020, and MeSH search terms were specific for each topic developed through the 7 chapters. The authors rated the quality of evidence and strength of recommendations using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. See online Appendix A (Tables A1 for definitions and A2 for interpretations of strong and weak recommendations). INTENDED AUDIENCE: physicians, including gynaecologists, obstetricians, family physicians, internists, emergency medicine specialists; nurses, including registered nurses and nurse practitioners; pharmacists; medical trainees, including medical students, residents, fellows; and other providers of health care for the target population. RECOMMENDATIONS.


The effect of acute caffeine ingestion on upper and lower body anaerobic exercise performance.
The current study examined the effect of acute caffeine ingestion on mean and peak power production, fatigue index and rating of perceived exertion (RPE) during upper body and lower body Wingate anaerobic test (WANT) performance. Using a double-blind design, 22 males undertook one upper body and one lower body WANT, 60 min following ingestion of caffeine (5 mg*kg-1) and one upper body and one lower body WANT following ingestion of placebo (5 mg*kg-1 Dextrose). Peak power was significantly higher (P = .001) following caffeine ingestion in both upper and lower body WANT. Peak power and mean power was also significantly higher during lower body, compared to upper body WANTs irrespective of substance ingested. However, caffeine ingestion did not enhance mean power neither in upper nor lower-body WANT. There were no significant differences in mean fatigue index as a consequence of substance ingested or mode of exercise (all P > 0.05). For RPE there was also a significant substance ingested X mode interaction (P = .001) where there were no differences in RPE between caffeine and placebo conditions in lower body WANTs but significantly lower RPE during upper body WANT in the presence of caffeine compared to placebo (P = .014). This is the first study to compare the effects of caffeine ingestion on upper and lower body 30-second WANT performance and suggests that caffeine ingestion in the dose of 5 mg*kg-1 ingested 60 min prior to exercise significantly enhances peak power when data from upper and lower body WANTs are combined.


Deep sequencing reveals mixed infection with 2009 pandemic influenza A (H1N1) virus strains and the emergence of oseltamivir resistance.
Mixed infections with seasonal influenza A virus strains are a common occurrence and an important source of genetic diversity. Prolonged viral shedding, as observed in immunocompromised individuals, can lead to mutational accumulation over extended periods. Recently, drug resistance was reported in immunosuppressed patients infected with the 2009 pandemic influenza A (H1N1) virus within a few days after oseltamivir treatment was initiated. To better understand the evolution and emergence of drug resistance in these circumstances, we used a deep sequencing approach to survey the viral population from an immunosuppressed patient infected with H1N1/2009 influenza and treated with neuraminidase inhibitors. This patient harbored 3 genetic variants from 2 phylogenetically distinct viral clades of pandemic H1N1/2009, strongly suggestive of mixed infection. Strikingly, one of these variants also developed drug resistance de novo in response to oseltamivir treatment. Immunocompromised individuals may, therefore, constitute an important source of genetic and phenotypic diversity, both through mixed infection and de novo mutation.


Quantifying the biomass of parasites to understand their role in aquatic communities.
By infecting multiple host species and acting as a food resource, parasites can affect food web topography and contribute to ecosystem energy transfer. Owing to the remarkable secondary production of some taxa, parasite biomass - although cryptic - can be comparable to other invertebrate and vertebrate groups. More resolved estimates of parasite biomass are therefore needed to understand parasite interactions, their consequences for host fitness, and potential influences on ecosystem energetics. We developed an approach to quantify the masses of helminth parasites and compared our results with those of biovolume-based approaches. Specifically, we massed larval and adult parasites representing 13 species and five life stages of trematodes and cestodes from snail and amphibian hosts. We used a replicated regression approach to quantify dry mass and compared these values with indirect biovolume estimates to test the validity of density assumptions. Our technique provided precise estimates (R (2) from 0.69 to 0.98) of biomass across a wide range of parasite morphotypes and sizes. Individual parasites ranged in mass from 0.368 ± 0.041 to 320 ± 98.1 μg. Among trematodes, adult parasites tended to be the largest followed by rediae, with nonclonal larval stages (metacercariae and cercariae) as the smallest. Among similar morphotypes, direct estimates of dry mass and the traditional biovolume technique provided generally comparable estimates (although important exceptions also emerged). Finally, we present generalized length-mass regression equations to calculate trematode mass from length measurements, and discuss the most efficient use of limited numbers of parasites. By providing a novel method of directly estimating parasite biomass while also helping to validate more traditional methods involving length-mass conversion, our findings aim to facilitate future investigations into the ecological significance of parasites, particularly with respect to ecosystem energetics. In addition, this novel technique can be applied to a wide range of difficult-to-mass organisms.


[The treatment of lupus nephritis, between consolidated strategies and new therapeutic options: a narrative review].
Over a half of patients with Systemic Lupus Erythematosus will develop lupus nephritis (LN). The diagnosis of LN, suspected based on clinical data (proteinuria, active urinary sediment, renal dysfunction), is confirmed with renal biopsy. The immunosuppressive treatment of proliferative classes of LN is based on an induction phase, where high-dose steroids are used in conjunction with mycophenolate mofetil (MMF) or cyclophosphamide, and a subsequent maintenance phase, that combines low-dose steroids with MMF or azathioprine. Different classes of drugs (calcineurin inhibitors, anti-CD20) can be used as an alternative, or in resistant forms of LN, although their role is less well-established. Recently published (or nearing completion) studies have opened up the possibility of using new drugs in LN. In particular, depletion (Obinutuzumab, anti-CD20 monoclonal antibody) or neutralization (Belimumab, anti-"B-cell activating factor" monoclonal antibody) of B lymphocytes, and the use of a calcineurin inhibitor with a low profile of renal and systemic toxicity (Voclosporin) demonstrated an improvement in renal response in addition to standard therapy.