Thrombosis, Bleeding, and the Observational Effect of Early Therapeutic Anticoagulation on Survival in Critically Ill Patients With COVID-19.
Hypercoagulability may be a key mechanism of death in patients with coronavirus disease 2019 (COVID-19). To evaluate the incidence of venous thromboembolism (VTE) and major bleeding in critically ill patients with COVID-19 and examine the observational effect of early therapeutic anticoagulation on survival. In a multicenter cohort study of 3239 critically ill adults with COVID-19, the incidence of VTE and major bleeding within 14 days after intensive care unit (ICU) admission was evaluated. A target trial emulation in which patients were categorized according to receipt or no receipt of therapeutic anticoagulation in the first 2 days of ICU admission was done to examine the observational effect of early therapeutic anticoagulation on survival. A Cox model with inverse probability weighting to adjust for confounding was used. 67 hospitals in the United States. Adults with COVID-19 admitted to a participating ICU. Time to death, censored at hospital discharge, or date of last follow-up. Among the 3239 patients included, the median age was 61 years (interquartile range, 53 to 71 years), and 2088 (64.5%) were men. A total of 204 patients (6.3%) developed VTE, and 90 patients (2.8%) developed a major bleeding event. Independent predictors of VTE were male sex and higher D-dimer level on ICU admission. Among the 2809 patients included in the target trial emulation, 384 (11.9%) received early therapeutic anticoagulation. In the primary analysis, during a median follow-up of 27 days, patients who received early therapeutic anticoagulation had a similar risk for death as those who did not (hazard ratio, 1.12 [95% CI, 0.92 to 1.35]). Observational design. Among critically ill adults with COVID-19, early therapeutic anticoagulation did not affect survival in the target trial emulation. None.


Long-term consequences of Kawasaki disease. A 10- to 21-year follow-up study of 594 patients.
The long-term consequences of the cardiovascular sequelae in Kawasaki disease remain uncertain. We identified 594 consecutive children with acute Kawasaki disease between 1973 and 1983, and this cohort was followed up for 10 to 21 years (mean, 13.6 years). In all patients, we evaluated coronary lesions by coronary angiography just after the acute stage. One hundred and forty-six patients (24.6%) were diagnosed as having coronary aneurysms. A second angiogram was performed 1 to 2 years later in all 146 patients who previously had coronary aneurysms, which demonstrated that 72 (49.3%) of these 146 had regression in the coronary aneurysm. A third angiogram was performed for 62 patients, a fourth for 29, and a fifth for 17. By 10 to 21 years after the onset of the illness, stenosis in the coronary aneurysm had developed in 28 patients. Myocardial infarction occurred in 11 patients, 5 of whom died. In the 26 patients with giant coronary aneurysms, stenotic lesions developed in 12, and no regression occurred. The 448 patients with normal findings at the first angiogram subsequently never developed any abnormal cardiac findings. Systemic artery aneurysms developed in 13 patients (2.2%), and valvular heart disease appeared in 7 (1.2%). The incidence of coronary aneurysm in acute Kawasaki disease was 25%, 55% of which showed regression. During follow-up, ischemic heart disease developed in 4.7% and myocardial infarction in 1.9%. Death occurred in 0.8%.


Diabetes in pancreatectomized baboons: a model for pancreatic transplantation studies.
This study was designed to assess plasma glucose levels (PLG) and insulin release in totally pancreatectomized baboons when challenged with intravenous glucose administration (IVGTT). Ten animals (Papio ursinus) were used and duodenectomy was intentionally avoided. The PLG at death was 18.0 +/- mmol/L, and the mean K-value within 3 days after pancreatectomy was 0.4% +/- 0.2%, indicating a significant impairment of glucose disappearance from the blood when compared to the control animals (P less than 0.01). Plasma insulin levels before and after stimulation with glucose were below the lowest level of insulin assay sensitivity. We conclude that in the primate, as in the dog, surgical pancreatectomy produced a reliable diabetic model, which is uniformly lethal if left untreated.


Dasatinib induces lung vascular toxicity and predisposes to pulmonary hypertension.
Pulmonary arterial hypertension (PAH) is a life-threatening disease that can be induced by dasatinib, a dual Src and BCR-ABL tyrosine kinase inhibitor that is used to treat chronic myelogenous leukemia (CML). Today, key questions remain regarding the mechanisms involved in the long-term development of dasatinib-induced PAH. Here, we demonstrated that chronic dasatinib therapy causes pulmonary endothelial damage in humans and rodents. We found that dasatinib treatment attenuated hypoxic pulmonary vasoconstriction responses and increased susceptibility to experimental pulmonary hypertension (PH) in rats, but these effects were absent in rats treated with imatinib, another BCR-ABL tyrosine kinase inhibitor. Furthermore, dasatinib treatment induced pulmonary endothelial cell apoptosis in a dose-dependent manner, while imatinib did not. Dasatinib treatment mediated endothelial cell dysfunction via increased production of ROS that was independent of Src family kinases. Consistent with these findings, we observed elevations in markers of endothelial dysfunction and vascular damage in the serum of CML patients who were treated with dasatinib, compared with CML patients treated with imatinib. Taken together, our findings indicate that dasatinib causes pulmonary vascular damage, induction of ER stress, and mitochondrial ROS production, which leads to increased susceptibility to PH development.


Strong or Weak Handgrip? Normative Reference Values for the German Population across the Life Course Stratified by Sex, Age, and Body Height.
Handgrip strength is an important biomarker of healthy ageing and a powerful predictor of future morbidity and mortality both in younger and older populations. Therefore, the measurement of handgrip strength is increasingly used as a simple but efficient screening tool for health vulnerability. This study presents normative reference values for handgrip strength in Germany for use in research and clinical practice. It is the first study to provide normative data across the life course that is stratified by sex, age, and body height. The study used a nationally representative sample of test participants ages 17-90. It was based on pooled data from five waves of the German Socio-Economic Panel (2006-2014) and involved a total of 11,790 persons living in Germany (providing 25,285 observations). Handgrip strength was measured with a Smedley dynamometer. Results showed that peak mean values of handgrip strength are reached in men's and women's 30s and 40s after which handgrip strength declines in linear fashion with age. Following published recommendations, the study used a cut-off at 2 SD below the sex-specific peak mean value across the life course to define a 'weak grip'. Less than 10% of women and men aged 65-69 were classified as weak according to this definition, shares increasing to about half of the population aged 80-90. Based on survival analysis that linked handgrip strength to a relevant outcome, however, a 'critically weak grip' that warrants further examination was estimated to commence already at 1 SD below the group-specific mean value.


A 3.3 Å-Resolution Structure of Hyperthermophilic Respiratory Complex III Reveals the Mechanism of Its Thermal Stability.
Respiratory chain complexes convert energy by coupling electron flow to transmembrane proton translocation. Owing to a lack of atomic structures of cytochrome bc1 complex (Complex III) from thermophilic bacteria, little is known about the adaptations of this macromolecular machine to hyperthermophilic environments. In this study, we purified the cytochrome bc1 complex of Aquifex aeolicus, one of the most extreme thermophilic bacteria known, and determined its structure with and without an inhibitor at 3.3 Å resolution. Several residues unique for thermophilic bacteria were detected that provide additional stabilization for the structure. An extra transmembrane helix at the N-terminus of cyt. c1 was found to greatly enhance the interaction between cyt. b and cyt. c1 , and to bind a phospholipid molecule to stabilize the complex in the membrane. These results provide the structural basis for the hyperstability of the cytochrome bc1 complex in an extreme thermal environment.


Predicting the ligand-binding properties of Borrelia burgdorferi s.s. Bmp proteins in light of the conserved features of related Borrelia proteins.
Bacteria of the genus Borrelia cause vector-borne infections like the most important hard tick-borne disease in the northern hemisphere, Lyme borreliosis (LB), and soft tick or louse transmitted relapsing fevers (RF), prevalent in temperate and tropical areas. Borrelia burgdorferi sensu lato (s.l.) includes several genospecies and causes LB in humans. In infected patients, Borrelia burgdorferi sensu stricto (s.s.) expresses the BmpA, BmpB, BmpC and BmpD proteins. The role of these proteins in the pathogenesis of LB remains incompletely characterized, but they are, however, closely related to Treponema pallidum PnrA (Purine nucleoside receptor A), a substrate-binding lipoprotein of the ATP-binding cassette (ABC) transporter family preferentially binding purine nucleosides. Based on 3D homology modeling, the Bmp proteins share the typical fold of the substrate-binding protein family and the ligand-binding properties of BmpA, BmpB and BmpD are highly similar, whereas those of BmpC differ markedly. Nevertheless, these residues are highly conserved within the genus Borrelia and the inferred phylogenetic tree also reveals that the RF Borrelia lack BmpB proteins but has an additional Bmp protein (BmpA2) missing in LB-causing Borrelia burgdorferi s.l. Our results indicate that the Bmp proteins could bind nucleosides, although BmpC might have a different ligand-binding specificity and, therefore, a distinct function. Furthermore, the work provides a means for classifying the Bmp proteins and supports further elucidation of the roles of these proteins.


Multicenter Registry of Real-World Patients With Severely Calcified Coronary Lesions Undergoing Orbital Atherectomy: 1-Year Outcomes.
We report the 1-year outcomes of real-world patients with severely calcified coronary arteries who underwent orbital atherectomy. Percutaneous coronary intervention of heavily calcified lesions is technically challenging and associated with worse clinical outcomes. Modification of severely calcified coronary lesions with orbital atherectomy facilitates stent delivery and expansion. Although we previously reported the safety of orbital atherectomy at 30 days in all comers with severely calcified coronary lesions, including patients who were excluded from the ORBIT II trial, longer-term follow-up is unknown. We retrospectively analyzed 458 all-comer patients who underwent orbital atherectomy followed by stenting from October 2013 to December 2015 at three centers. The primary endpoint was the 1-year major adverse cardiac and cerebrovascular event (MACCE) rate, defined as the composite of death, myocardial infarction, target-vessel revascularization, and stroke. One-year data were available for 453/457 patients (98.9%). At 1-year follow-up, the MACCE rate was 12.6%, death rate was 4.0%, myocardial infarction rate was 1.8%, target-vessel revascularization rate was 7.5%, stroke rate was 1.3%, and stent thrombosis rate was 1.3%. Orbital atherectomy is a valuable option for the treatment of severely calcified coronary arteries, including patients with very complex coronary anatomy and severe underlying comorbid conditions. Orbital atherectomy provided acceptable outcomes at 1 year and compared favorably to historical controls. A randomized trial with longer follow-up is needed to determine the optimal treatment strategy for patients with severely calcified coronary lesions.


Brachycephalic feline noses: CT and anatomical study of the relationship between head conformation and the nasolacrimal drainage system.
A study was designed to evaluate the influence of head conformation on the course of the nasolacrimal drainage system (NDS) in 31 brachycephalic and 15 mesocephalic cats using computed tomography (CT), CT-dacryocystography and anatomical methods. The higher the degree of brachycephalia, the more the facial bones and upper canine teeth are displaced dorsally (ie, the more pronounced the dorsorotation). Dorsorotation leads to abnormal dislocation of the ventral nasal concha and to almost horizontally rotated upper canine teeth, and thus a steeply oriented NDS. In severe brachycephalia the NDS is forced to pass below the canine tooth (adopt a V-shaped course) and the drainage function seems to be inefficient. The rotation of the upper canine teeth appears to provide a basis for classification of brachycephalia in cats. The authors recommend that breeders avoid breeding from individuals affected by this condition and to give preference to cats with longer facial bones.


Tetrahedral colloidal clusters from random parking of bidisperse spheres.
Using experiments and simulations, we investigate the clusters that form when colloidal spheres stick irreversibly to--or "park" on--smaller spheres. We use either oppositely charged particles or particles labeled with complementary DNA sequences, and we vary the ratio α of large to small sphere radii. Once bound, the large spheres cannot rearrange, and thus the clusters do not form dense or symmetric packings. Nevertheless, this stochastic aggregation process yields a remarkably narrow distribution of clusters with nearly 90% tetrahedra at α = 2.45. The high yield of tetrahedra, which reaches 100% in simulations at α = 2.41, arises not simply because of packing constraints, but also because of the existence of a long-time lower bound that we call the "minimum parking" number. We derive this lower bound from solutions to the classic mathematical problem of spherical covering, and we show that there is a critical size ratio α(c) = (1 + sqrt[2]) ≈ 2.41, close to the observed point of maximum yield, where the lower bound equals the upper bound set by packing constraints. The emergence of a critical value in a random aggregation process offers a robust method to assemble uniform clusters for a variety of applications, including metamaterials.


Petiveria alliacea L. extract protects mice against Listeria monocytogenes infection--effects on bone marrow progenitor cells.
In this study we have investigated the effects of Petiveria alliacea on the hematopoietic response of mice infected with Listeria monocytogenes. Our results demonstrate a protective effect of the crude extract of P. alliacea since the survival of the treated/infected was higher than that in the infected group. Moreover, the number of granulocyte/macrophage colonies (CFU-GM) and the serum colony stimulating activity levels were increased in the treated/infected mice in relation to the infected group. These results suggest an immunomodulation of Petiveria alliacea extract on hematopoiesis, which may be responsible, at least in part, for the increased resistance of mice to Listeria monocytogenes infection.


Chemical spraying as reported by refugees from South Vietnam.
Ninety-eight refugees who had been exposed to chemical sprays in South Vietnam were interviewed in Hanoi. Most reported effects on eyes and skin and gastrointestinal upsets. Ninety-two percent suffered fatigue, prolonged or indefinite in 17 percent of cases. Reports of abortions and monstrous births in sprayed humans and animals and of substantial numbers of deaths among fish, fowl, and pigs were also given.


On-chip cell lysis by antibacterial non-leaching reusable quaternary ammonium monolithic column.
Reusable antibacterial non-leaching monolithic columns polymerized in microfluidic channels designed for on-chip cell lysis applications were obtained by the photoinitiated free radical copolymerization of diallyldimethylammonium chloride (DADMAC) and ethylene glycol diacrylate (EGDA) in the presence of a porogenic solvent. The microfluidic channels were fabricated in cross-linked poly(methyl methacrylate) (X-PMMA) substrates by laser micromachining. The monolithic columns have the ability to inhibit the growth of, kill and efficiently lyse Gram-positive Micrococcus luteus (Schroeter) (ATCC 4698) and Kocuria rosea (ATCC 186), and Gram-negative bacteria Pseudomonas putida (ATCC 12633) and Escherichia coli (ATCC 35218) by mechanically shearing the bacterial membrane when forcing the cells to pass through the narrow pores of the monolithic column, and simultaneously disintegrating the cell membrane by physical contact with the antibacterial surface of the column. Cell lysis was confirmed by off-chip PCR without the need for further purification. The influence of the cross-linking monomer on bacterial growth inhibition, leaching, lysis efficiency of the monolithic column and its mechanical stability within the microfluidic channel were investigated and analyzed for three different cross-linking monomers: ethylene glycol dimethacrylate (EGDA), ethylene glycol dimethacrylate (EGDMA) and 1,6-hexanediol dimethacrylate (1,6-HDDMA). Furthermore, the bonding efficiency of two X-PMMA substrates with different cross-linking levels was studied. The monolithic columns were shown to be stable, non-leaching, and reusable for over 30 lysis cycles without significant performance degradation or DNA carryover when they were back-flushed between lysis cycles.


The emotional eyewitness: the effects of emotion on specific aspects of eyewitness recall and recognition performance.
The present set of experiments aimed to investigate the effects of negative emotion on specific aspects of eyewitness recall and recognition performance. The experience of emotion was manipulated between subjects, with participants either viewing a crime scenario (a mugging) or a neutral scenario (a conversation). Eyewitness recall was categorized into descriptions of the perpetrator, critical incident, victim, and environmental details. The completeness and accuracy of eyewitness recall across categories of detail were measured in Experiment 1. A significant main effect of negative emotion was found for the completeness of recall. Furthermore, a significant main effect of the completeness of eyewitness statements was found, but not for their accuracy. However, these main effects were qualified by a significant interaction between emotion and category of detail recalled. Specifically, emotional participants provided a more complete description of the perpetrator than neutral participants; however, they were less able than their neutral counterparts to describe what the perpetrator did to the victim. In light of these findings, Experiment 2 investigated whether enhanced completeness of perpetrator descriptions during recall translated into an enhanced ability to recognize the perpetrator from a photographic lineup by emotional compared with neutral participants. Results from Experiment 2 suggest that while emotional participants again provide a more complete description of the perpetrator, they are less able than their neutral counterparts to recognize the perpetrator from a photographic lineup. Results are discussed in terms of a retrieval motivation hypothesis of negative emotional experience and the possible consequences for eyewitness testimony.


Mental health, coping and resilience among young men who have sex with men in Zambia.
Current law in Zambia criminalises same-sex sexual contact while strong socio-cultural values drive a profoundly negative view of the moral status of gender and sexual minorities. Despite this, Zambia's national HIV programme has recently identified the predominantly young population of men who have sex with men as a priority group for HIV and other sexual health programming. Research in other African settings has shown how the sexual health of these young men is affected by mental health. This mixed-methods study, which drew on the minority stress model as an analytical framework, sought to explore mental health as an initial step towards understanding its influence on other health domains. Findings describe the tension, and its psychological effects, surrounding the trajectory of discovering and affirming same-sex sexuality in an environment replete with social, physical and emotional risks, but one in which young men must nevertheless seek to create and maintain a meaningful, if precarious, social existence. To achieve this, in the absence of supportive mental health services or other programmatic responses, they adopt numerous risk-mitigation or coping strategies to attempt to build resiliency and to preserve their psychological and emotional well-being.


Causes of male sexual trait divergence in introduced populations of guppies.
Males from different populations of the same species often differ in their sexually selected traits. Variation in sexually selected traits can be attributed to sexual selection if phenotypic divergence matches the direction of sexual selection gradients among populations. However, phenotypic divergence of sexually selected traits may also be influenced by other factors, such as natural selection and genetic constraints. Here, we document differences in male sexual traits among six introduced Australian populations of guppies and untangle the forces driving divergence in these sexually selected traits. Using an experimental approach, we found that male size, area of orange coloration, number of sperm per ejaculate and linear sexual selection gradients for male traits differed among populations. Within populations, a large mismatch between the direction of selection and male traits suggests that constraints may be important in preventing male traits from evolving in the direction of selection. Among populations, however, variation in sexual selection explained more than half of the differences in trait variation, suggesting that, despite within-population constraints, sexual selection has contributed to population divergence of male traits. Differences in sexual traits were also associated with predation risk and neutral genetic distance. Our study highlights the importance of sexual selection in trait divergence in introduced populations, despite the presence of constraining factors such as predation risk and evolutionary history.


Plasma triglyceride level and mortality from coronary heart disease.
Whether the plasma triglyceride level is a risk factor for coronary heart disease has been controversial, and evaluation of the triglyceride level as a risk factor is fraught with methodologic difficulties. We studied the association between plasma triglyceride levels and the 12-year incidence of death from coronary heart disease in 10 North American populations participating in the Lipid Research Clinics Follow-up Study, while adjusting for the potential confounding effects of other risk factors for cardiovascular disease, including the level of high-density lipoprotein (HDL) cholesterol. All analyses were sex-specific, and separate analyses were performed in high and low strata of HDL cholesterol, low-density lipoprotein (LDL) cholesterol, fasting plasma glucose, and age. The rates of coronary death in both men and women increased with the triglyceride level. In Cox proportional-hazards models adjusted for age, in which the natural log of the triglyceride levels was used to give a normal distribution, the relative risk per natural-log unit of triglyceride (e.g., a triglyceride level of 150 mg per deciliter vs. a level of 55 mg per deciliter) was 1.54 (95 percent confidence interval, 1.19 to 1.98; P < 0.001) in men and 1.88 (95 percent confidence interval, 1.19 to 2.98; P < 0.007) in women. After an adjustment for potential covariates, however, these relative risks were not statistically significant. Analyses based on lipoprotein cholesterol levels revealed a positive association between the triglyceride level and coronary mortality in the lower stratum of both HDL and LDL cholesterol, but not in the higher stratum. Conversely, the HDL cholesterol level was unrelated to coronary mortality in the lower stratum of LDL cholesterol, but was strongly inversely associated with coronary death in the higher stratum of LDL cholesterol. The relative risk of coronary death associated with triglyceride level was higher at younger ages. The associations between the triglyceride level and coronary mortality in the lower HDL cholesterol, LDL cholesterol, and age strata were small and were further reduced by an adjustment for the fasting plasma glucose level. Overall, the plasma triglyceride level showed no independent association with coronary mortality. However, in subgroups of subjects with lower HDL and LDL cholesterol levels and in younger subjects, defined a priori, an association between the triglyceride level and coronary mortality was observed, although this association was small and was not statistically significant after an adjustment for the plasma glucose level.


High-Dose, Diazoxide-Mediated Insulin Suppression Boosts Weight Loss Induced by Lifestyle Intervention.
Obesity-related hyperinsulinism may impede lifestyle-initiated weight loss. Proof-of-concept study to investigate the amplifying effects of diazoxide (DZX)-mediated insulin suppression on lifestyle-induced weight loss in nondiabetic, hyperinsulinemic, obese men. Twelve-month study comprising an initial 6-month, double-blind trial, followed by a partially de-blinded 6-month extension in men with obesity with a body mass index of 30 to 37.5 kg/m2 and a fasting serum C-peptide level >1.00 nM. Patients were randomized into three treatment groups: DZX + placebo (DZX + PL), DZX + metformin (DZX + MTF), and double PL (PL + PL). At 6 months, DZX treatment was associated with a 6.1-kg PL-subtracted decline in fat mass (FM), and at 12 months, FM had decreased by a total of 15.7 ± 2.5 kg. Twelve months of DZX treatment was also associated with a significant decline in systolic (-6.6%) and diastolic (-8.6%) blood pressure and low-density lipoprotein-cholesterol (-18%) and triglycerides (-43%) and a 39% rise in high-density lipoprotein-cholesterol. These effects were achieved at the cost of a small rise in fasting glucose (95% CI: 0.2 to 1.0 mM) and hemoglobin A1c (95% CI: -0.08% to 0.44%). There were no differences between DZX monotherapy and the combination of DZX + MTF. High-dose DZX treatment of 1 year resulted in a substantial decrease in FM, blood pressure, and lipid levels at the cost of a small rise in blood glucose levels.


Respiratory disease in cotton textile workers in the People's Republic of China. II. Pulmonary function results.
Pulmonary function tests were performed pre and post workshift on 887 textile workers with at least two years of employment in two cotton mills and one silk mill in Shanghai, the People's Republic of China. Environmental sampling was performed with vertical elutriators, and pulmonary function was performed with standardized techniques. Cotton textile workers were found to have greater across-shift decrements in forced expiratory volume in 1 s (FEV1.0) than silk workers. Increasing duration of exposure resulted in increasing acute decrements in FEV1.0, although significant acute decrements were found in workers with less than five years of exposure. The acute changes in FEV 1.0 were noted in both symptomatic and asymptomatic cotton workers, though the difference between the across-shift change in FEV1.0 (delta FEV1.0%) of the byssinotics and nonbyssinotics increased as work duration increased. There was no difference in preshift FEV1.0 between the cotton and silk workers, but several selection factors likely influenced the observations.


Comparative research on phosphorus removal by pilot-scale vertical flow constructed wetlands using steel slag and modified steel slag as substrates.
This research mainly focused on the phosphorus removal performance of pilot-scale vertical flow constructed wetlands with steel slag (SS) and modified steel slag (MSS). First, bench-scale experiments were conducted to evaluate the phosphorus adsorption capacity. Results showed that the Langmuir model could better describe the adsorption characteristics of the two materials; the maximum adsorption of MSS reached 12.7 mg/g, increasing by 34% compared to SS (9.5 mg/g). Moreover, pilot-scale constructed wetlands with SS and MSS were set up outdoors. Then, the influence of hydraulic retention time (HRT) and phosphorus concentration in phosphorus removal for two wetlands were investigated. Results revealed that better performance of the two systems could be achieved with an HRT of 2 d and phosphorus concentration in the range of 3-4.5 mg/L; the system with MSS had a better removal efficiency than the one with SS in the same control operation. Finally, the study implied that MSS could be used as a promising substrate for wetlands to treat wastewater with a high phosphorus concentration. However, considering energy consumption, SS could be regarded as a better alternative for substrate when treating sewage with a low phosphorus concentration.


Global disparities in SARS-CoV-2 genomic surveillance.
Genomic sequencing is essential to track the evolution and spread of SARS-CoV-2, optimize molecular tests, treatments, vaccines, and guide public health responses. To investigate the global SARS-CoV-2 genomic surveillance, we used sequences shared via GISAID to estimate the impact of sequencing intensity and turnaround times on variant detection in 189 countries. In the first two years of the pandemic, 78% of high-income countries sequenced >0.5% of their COVID-19 cases, while 42% of low- and middle-income countries reached that mark. Around 25% of the genomes from high income countries were submitted within 21 days, a pattern observed in 5% of the genomes from low- and middle-income countries. We found that sequencing around 0.5% of the cases, with a turnaround time <21 days, could provide a benchmark for SARS-CoV-2 genomic surveillance. Socioeconomic inequalities undermine the global pandemic preparedness, and efforts must be made to support low- and middle-income countries improve their local sequencing capacity.


4-protein signature predicting tamoxifen treatment outcome in recurrent breast cancer.
Estrogen receptor (ER) positive tumors represent the majority of breast malignancies, and are effectively treated with hormonal therapies, such as tamoxifen. However, in the recurrent disease resistance to tamoxifen therapy is common and a major cause of death. In recent years, in-depth proteome analyses have enabled identification of clinically useful biomarkers, particularly, when heterogeneity in complex tumor tissue was reduced using laser capture microdissection (LCM). In the current study, we performed high resolution proteomic analysis on two cohorts of ER positive breast tumors derived from patients who either manifested good or poor outcome to tamoxifen treatment upon recurrence. A total of 112 fresh frozen tumors were collected from multiple medical centers and divided into two sets: an in-house training and a multi-center test set. Epithelial tumor cells were enriched with LCM and analyzed by nano-LC Orbitrap mass spectrometry (MS), which yielded >3000 and >4000 quantified proteins in the training and test sets, respectively. Raw data are available via ProteomeXchange with identifiers PXD000484 and PXD000485. Statistical analysis showed differential abundance of 99 proteins, of which a subset of 4 proteins was selected through a multivariate step-down to develop a predictor for tamoxifen treatment outcome. The 4-protein signature significantly predicted poor outcome patients in the test set, independent of predictive histopathological characteristics (hazard ratio [HR] = 2.17; 95% confidence interval [CI] = 1.15 to 4.17; multivariate Cox regression p value = 0.017). Immunohistochemical (IHC) staining of PDCD4, one of the signature proteins, on an independent set of formalin-fixed paraffin-embedded tumor tissues provided and independent technical validation (HR = 0.72; 95% CI = 0.57 to 0.92; multivariate Cox regression p value = 0.009). We hereby report the first validated protein predictor for tamoxifen treatment outcome in recurrent ER-positive breast cancer. IHC further showed that PDCD4 is an independent marker.


Periosteal osteosarcoma--a European review of outcome.
Periosteal osteosarcoma is a rare primary malignant bone tumour. Treatment is by surgical excision, but controversy remains about the value of chemotherapy. The members of the European Musculo Skeletal Oncology Society (EMSOS) collaborated to produce a dataset of 119 patients. The predominant site for the tumour was the femur, followed by the tibia. All but 2 patients underwent surgery, with 9 requiring amputation and the others having limb salvage. A total of 81 patients had chemotherapy, of whom 50 had neoadjuvant chemotherapy. There was no standard chemotherapy regime, but all patients receiving chemotherapy were given doxorubicin combined with at least one other agent. The overall survival was 89% at 5 years and 83% at 10 years. Eight patients developed local recurrence, of whom 5 died. Survival was related to appearance of local recurrence (P < 0.0001) but no other single factor. The use of chemotherapy was not shown to be a prognostic factor, but was used in two-thirds of the patients in this study.


A biomechanical analysis of the stand-up paddle board stroke: a comparative study.
Stand-up paddle boarding (SUP) is a rapidly growing global aquatic sport, with increasing popularity among participants within recreation, competition and rehabilitation. To date, few scientific studies have focused on SUP. Further, there is no research examining the biomechanics of the SUP paddle stroke. The purpose of this study was to investigate whether variations in kinematics existed among experienced and inexperienced SUP participants using three-dimensional motion analysis. This data could be of significance to participants, researchers, coaches and health practitioners to improve performance and inform injury minimization strategies. A cross-sectional observational design study was performed with seven experienced and 19 inexperienced paddlers whereby whole-body kinematic data were acquired using a six-camera Vicon motion capture system. Participants paddled on a SUP ergometer while three-dimensional range of motion (ROM) and peak joint angles were calculated for the shoulders, elbows, hips and trunk. Mann-Whitney U tests were conducted on the non-normally distributed data to evaluate differences between level of expertise. Significant differences in joint kinematics were found between experienced and inexperienced participants, with inexperienced participants using greater overall shoulder ROM (78.9° ± 24.9° vs 56.6° ± 17.3°, p = 0.010) and less hip ROM than the experienced participants (50.0° ± 18.5° vs 66.4° ± 11.8°, p = 0.035). Experienced participants demonstrated increased shoulder motion at the end of the paddle stoke compared to the inexperienced participants (74.9° ± 16.3° vs 35.2° ± 28.5°, p = 0.001 minimum shoulder flexion) and more extension at the elbow (6.0° ± 9.2° minimum elbow flexion vs 24.8° ± 13.5°, p = 0.000) than the inexperienced participants. The results of this study indicate several significant kinematic differences between the experienced and inexperienced SUP participants. These variations in technique were noted in the shoulder, elbow and hip and are evident in other aquatic paddling sports where injury rates are higher in these joints. These finding may be valuable for coaches, therapists and participants needing to maximize performance and minimize injury risk during participation in SUP.


Alterations in body composition in Indian patients with non-small cell lung cancer.
Alterations in body composition are common in cancer and may affect outcomes differentially based on geographical and ethnic factors. However, data in lung cancer are sparse and conflicting. We compared the body composition of Indian lung cancer patients with healthy subjects using a retrospective review of all newly diagnosed patients with nonsmall cell lung cancer. Age- and sex-matched healthy controls were recruited prospectively. Basal metabolic rate (BMR), total body water (TBW), fat mass, and fat-free mass (FFM) were calculated by bioelectric impedance method. A total of 256 patients (83.6% males) and 210 controls (81.4% males) were studied. The mean (standard deviation) age of patients was 54.5 (9.0) years, median smoking index was 598.2 (range, 0-2500), and median Karnofsky performance scale (KPS) was 80 (range, 40-100). Majority (54.7%) had Stage IV disease. All components of body composition, i.e., BMR, TBW, fat mass, and FFM, were significantly lower (P < 0.01) in patients as compared to controls. Body mass index, fat mass, FFM, and TBW were lower in older subjects with poorer KPS. The presence of metastasis or symptom duration did not affect body composition. These results indicate that Indian patients with lung cancer have altered body composition which declines with increasing age and worsening performance status.


Development and Application of a Microfluidics-Based Panel in the Basal/Luminal Transcriptional Characterization of Archival Bladder Cancers.
In the age of personalized medicine stratifying tumors into molecularly defined subtypes associated with distinctive clinical behaviors and predictable responses to therapies holds tremendous value. Towards this end, we developed a custom microfluidics-based bladder cancer gene expression panel for characterization of archival clinical samples. In silico analysis indicated that the content of our panel was capable of accurately segregating bladder cancers from several public datasets into the clinically relevant basal and luminal subtypes. On a technical level, our bladder cancer panel yielded robust and reproducible results when analyzing formalin-fixed, paraffin-embedded (FFPE) tissues. We applied our panel in the analysis of a novel set of 204 FFPE samples that included non-muscle invasive bladder cancers (NMIBCs), muscle invasive disease (MIBCs), and bladder cancer metastases (METs). We found NMIBCs to be mostly luminal-like, MIBCs to include both luminal- and basal-like types, and METs to be predominantly of a basal-like transcriptional profile. Mutational analysis confirmed the expected enrichment of FGFR3 mutations in luminal samples, and, consistently, FGFR3 IHC showed high protein expression levels of the receptor in these tumors. Our bladder cancer panel enables basal/luminal characterization of FFPE tissues and with further development could be used for stratification of bladder cancer samples in the clinic.


Tracking of Pedometer-Determined Physical Activity in Healthy Elderly Japanese People.
Tracking refers to the tendency for an individual to maintain their rank within a group over time. This study longitudinally investigated the 8-year tracking of pedometer-determined physical activity among physically independent elderly Japanese people aged 72 to 80 years. Steps/day were measured when participants were aged 72 and assessed again after 2, 5, and 8 years. The number of participants with a 2-year follow-up (72 to 74 years) was 177 (101 males, 76 females), with a 5-year follow-up (72 to 77 years) was 183 people (110 males, 73 females), and with an 8-year follow-up (72 to 80 years) was 145 people (91 males, 54 females). Step counts were continuously measured for 1 week in January, April, July, and October during each year of assessment. A high rank correlation coefficient for steps/day exceeding 0.60 was obtained at the 2-, 5- and 8-year follow-up examinations for both males and females. This study suggests that the rank within a group of pedometer-determined steps/day remains stable over up to 8 years in healthy Japanese people aged between 72 to 80 years old.


Health services utilisation disparities between English speaking and non-English speaking background Australian infants.
To examine the differences in health services utilisation and the associated risk factors between infants from non-English speaking background (NESB) and English speaking background (ESB) within Australia. We analysed data from a national representative longitudinal study, the Longitudinal Study of Australian Children (LSAC) which started in 2004. We used survey logistic regression coupled with survey multiple linear regression to examine the factors associated with health services utilisation. Similar health status was observed between the two groups. In comparison to ESB infants, NESB infants were significantly less likely to use the following health services: maternal and child health centres or help lines (odds ratio [OR] 0.56; 95% confidence intervals [CI], 0.40-0.79); maternal and child health nurse visits (OR 0.68; 95% CI, 0.49-0.95); general practitioners (GPs) (OR 0.58; 95% CI, 0.40-0.83); and hospital outpatient clinics (OR 0.54; 95% CI, 0.31-0.93). Multivariate analysis results showed that the disparities could not be fully explained by the socioeconomic status and language barriers. The association between English proficiency and the service utilised was absent once the NESB was taken into account. Maternal characteristics, family size and income, private health insurance and region of residence were the key factors associated with health services utilisation. NESB infants accessed significantly less of the four most frequently used health services compared with ESB infants. Maternal characteristics and family socioeconomic status were linked to health services utilisation. The gaps in health services utilisation between NESB and ESB infants with regard to the use of maternal and child health centres or phone help, maternal and child health nurse visits, GPs and paediatricians require appropriate policy attentions and interventions.


Platelet-derived growth factor: a key factor in the pathogenesis of graves' ophthalmopathy and potential target for treatment.
Activation of orbital fibroblasts resulting in excessive proliferation, cytokine and hyaluronan production and differentiation into adipocytes, is a main determinant of orbital tissue inflammation and tissue expansion in Graves' ophthalmopathy (GO). During the last years we have shown that the platelet-derived growth factor (PDGF) isoforms PDGF-AA, PDGF-AB and PDGF-BB are increased in orbital tissue from GO patients with active and inactive disease. These PDGF isoforms exhibit the capacity to stimulate proliferation, hyaluronan and cytokine/chemokine production by orbital fibroblasts. Moreover, PDGF-AB and PDGF-BB increase thyroid stimulating hormone receptor (TSHR) expression by orbital fibroblasts, which enhances the orbital fibroblast activating capacity of the THSR stimulatory autoantibodies present in Graves' disease (GD) patients. Of these PDGF isoforms PDGF-BB exhibits the strongest orbital fibroblast activating effects, which is likely related to its ability to bind both the PDGF-receptor (PDGF-R)α and PDGF-Rβ chains. Thus the PDGF-system fulfills important roles in orbital fibroblast activation in both active and inactive GO, which supports a therapeutic rationale for blocking PDGF signaling in GO. Tyrosine kinase inhibitors (TKIs) may be candidates to target PDGF signaling. Of several TKIs tested dasatinib exhibited the highest potency to block PDGF-R signaling in orbital fibroblasts and may represent a promising compound for the treatment of GO as it was effective at low dosage and is associated with less side effects compared to imatinib mesylate and nilotinib. In this review the contribution of PDGF to the pathophysiology of GO as well as therapeutic approaches to target this PDGF-system will be addressed.


A century of cerebral achromatopsia.
This review is an enquiry into why the early clinical evidence for a colour centre in the cerebral cortex of man was so successfully dismissed for the best part of a century. The imperfection of this evidence cannot be the reason, for the same evidence that was rejected earlier is accepted today. Instead, it was because the prevalent concepts of vision as a function, and of the role of the cerebral cortex in it, dominated facts and prevented acceptance of evidence showing a specialization for colour in the visual cortex. It was only after those concepts were overthrown by the demonstration of functional specialization in the visual cortex of the primate that the evidence for a colour centre in the human brain became acceptable. Today, our new knowledge of the colour areas and pathways in the primate brain allows us to give a more complete account of the pathophysiology of cerebral achromatopsia in man.


Sex workers' accounts of condom use: implications for condom production, promotion and health policy.
To explore sex workers' accounts of condom use and their recommendations about how condoms might be improved. In-depth interviews were conducted with 22 female sex workers in sex work premises in London, UK and Amsterdam, The Netherlands. The consistent and effective use of condoms was dependent upon client selection, sex worker control of the condom, communication skills and on condom- and sex-related skills. The design of facilities, the way the encounter was structured and alarm call systems were key to generating an environment in which sex worker control of the interaction was feasible. A wide range of practices used for 'safer sex', including the use of simulated vaginal sex, skills in fitting the condom in a sexually arousing way, checking the condom placement during intercourse and holding onto the condom during withdrawal, were described but awareness of such practices was piecemeal. Several sex workers said that particular care is needed when using condoms in men with a small penis and pointed out to the authors that a smaller condom would be useful. The sex workers told us about the importance of environmental factors and a range of sex- and condom-related skills in 'safer sex'. Environmental 'safety' features could be addressed through a licensing system for sex work premises. Communication, condom- and sex-related skills should be more broadly disseminated through health promotion initiatives with sex workers. The issue of condom fit deserves further attention.


Climate change, migration and health systems resilience: Need for interdisciplinary research.
Climate change is one of today's major challenges, among the causes of population movements and international migration. Climate migrants impact health systems and how they respond and adapt to their needs and patterns. But to date, the resilience of health systems in the context of climate change has been little explored. The purpose of this article is to show the importance of studying, from an interdisciplinary perspective, the relationships between climate change, migration, and the resilience of health systems. Resilience is an old concept, notably in the field of psychology, and is increasingly applied to the study of health systems. Yet, no research has analysed the resilience of health systems in the context of climate change. While universal health coverage is a major international goal, little research has to date focused on the existing links between climate, migration, health systems and resilience. We propose an interdisciplinary approach relying on the concept of health system resilience to study adaptive and transformative strategies to articulate climate change, migration and health systems.


Cancer stem cells from colorectal cancer-derived cell lines.
Cancer stem cells (CSCs) are the subpopulation of cells within a tumor that can self-renew, differentiate into multiple lineages, and drive tumor growth. Here we describe a two-pronged approach for the identification and characterization of CSCs from colorectal cancer cell lines, using a Matrigel-based differentiation assay, and cell surface markers CD44 and CD24. About 20 to 30% of cells from the SW1222 cell line form megacolonies in Matrigel that have complex 3D structures resembling colonic crypts. The megacolonies' capacity to self-renew in vitro is direct evidence that they contain the CSCs. Furthermore, just 200 cells from SW1222 megacolonies initiate tumors in NOD/SCID mice. We also showed that CD44(+)CD24(+) cells enriched for colorectal CSCs in the HT29 and SW1222 cell lines, which can self-renew and reform all four CD44/CD24 subpopulations, are the most clonogenic in vitro and can initiate tumors in vivo. A single SW1222 CD44(+)CD24(+) CSC, when grown in Matrigel, can form large megacolonies that differentiate into enterocyte, enteroendocrine, and goblet cell lineages. The HCT116 line does not differentiate or express CDX1, nor does it contain subpopulations of cells with greater tumor-forming capacity, suggesting that HCT116 contains mainly CSCs. However, forced expression of CDX1 in HCT116 leads to reduced clonogenicity and production of differentiating crypt-containing colonies, which can explain the selection for reduced CDX1 expression in many colorectal cancers. In summary, colorectal cancer cell lines contain subpopulations of CSCs, characterized by their cell surface markers and colony morphology, which can self-renew and differentiate into multiple lineages.


An economic evaluation of universal vaccination against hepatitis B virus.
This report presents the results of an economic evaluation utilizing U.K. data, into a vaccination programme against Hepatitis B using a genetically engineered, yeast-derived vaccine, Engerix B. Cost-effectiveness ratios were calculated for four different programmes: an infant vaccination programme; a child vaccination programme; an adolescent vaccination programme; and a combined child and adolescent programme. For each programme, the number of annual cohorts vaccinated was varied from 1 to 25. The outcome was defined as incremental life years gained, and the results are reported as costs per incremental life-year gained. Benefits were calculated in both undiscounted and discounted forms. All costs were discounted. The discount rate used was 6%. All major epidemiological and cost assumptions were subjected to a sensitivity analysis. Baseline results with benefits discounted range from pounds188015 to pounds301365 per life year gained, depending on the duration of programme and vaccination strategy. With benefits undiscounted, the comparable range is from pounds5234 to pounds13034.


Gamma knife radiosurgery in the treatment of tumor-related facial pain.
Intracranial neoplasms can cause pain similar to trigeminal neuralgia. Literature regarding radiosurgery for this is limited. We present a retrospective review of patients with tumor-related facial pain from benign lesions treated with gamma knife radiosurgery (GKRS) at Wake Forest University. The primary objectives were to determine long-term pain relief and predictive factors for pain alleviation. We reviewed 515 patients treated with GKRS for benign meningioma, vestibular schwannoma or trigeminal schwannoma between August 1999 and August 2010. Twenty-one eligible patients had tumor-related facial pain prior to GKRS. The median marginal tumor dose was 12 Gy. Long-term pain relief data were obtained by chart review and telephone interview. The median follow-up for symptom evaluation was 3.8 years. Seventeen of 21 patients (81%) experienced a Barrow Neurological Institute (BNI) score of I-III at 6 months following GKRS. Kaplan-Meier estimates of freedom from BNI IV-V relapse were 66% at 1 year and 53% at 2 years. No pain relapses occurred after 2 years. GKRS of benign lesions is a noninvasive option for patients with tumor-related facial pain. Pain relief is modest, with the majority of pain relapses occurring within 2 years and approximately one half of patients maintaining relief beyond 2 years.


The role of oxidative stress in Parkinson's disease.
Oxidative stress plays an important role in the degeneration of dopaminergic neurons in Parkinson's disease (PD). Disruptions in the physiologic maintenance of the redox potential in neurons interfere with several biological processes, ultimately leading to cell death. Evidence has been developed for oxidative and nitrative damage to key cellular components in the PD substantia nigra. A number of sources and mechanisms for the generation of reactive oxygen species (ROS) are recognized including the metabolism of dopamine itself, mitochondrial dysfunction, iron, neuroinflammatory cells, calcium, and aging. PD causing gene products including DJ-1, PINK1, parkin, alpha-synuclein and LRRK2 also impact in complex ways mitochondrial function leading to exacerbation of ROS generation and susceptibility to oxidative stress. Additionally, cellular homeostatic processes including the ubiquitin-proteasome system and mitophagy are impacted by oxidative stress. It is apparent that the interplay between these various mechanisms contributes to neurodegeneration in PD as a feed forward scenario where primary insults lead to oxidative stress, which damages key cellular pathogenetic proteins that in turn cause more ROS production. Animal models of PD have yielded some insights into the molecular pathways of neuronal degeneration and highlighted previously unknown mechanisms by which oxidative stress contributes to PD. However, therapeutic attempts to target the general state of oxidative stress in clinical trials have failed to demonstrate an impact on disease progression. Recent knowledge gained about the specific mechanisms related to PD gene products that modulate ROS production and the response of neurons to stress may provide targeted new approaches towards neuroprotection.


The tyrosine phosphatase CD148 is an essential positive regulator of platelet activation and thrombosis.
Platelets play a fundamental role in hemostasis and thrombosis. They are also involved in pathologic conditions resulting from blocked blood vessels, including myocardial infarction and ischemic stroke. Platelet adhesion, activation, and aggregation at sites of vascular injury are regulated by a diverse repertoire of tyrosine kinase-linked and G protein-coupled receptors. Src family kinases (SFKs) play a central role in initiating and propagating signaling from several platelet surface receptors; however, the underlying mechanism of how SFK activity is regulated in platelets remains unclear. CD148 is the only receptor-like protein tyrosine phosphatase identified in platelets to date. In the present study, we show that mutant mice lacking CD148 exhibited a bleeding tendency and defective arterial thrombosis. Basal SFK activity was found to be markedly reduced in CD148-deficient platelets, resulting in a global hyporesponsiveness to agonists that signal through SFKs, including collagen and fibrinogen. G protein-coupled receptor responses to thrombin and other agonists were also marginally reduced. These results highlight CD148 as a global regulator of platelet activation and a novel antithrombotic drug target.


NG2/HMPG modulation of human articular chondrocyte adhesion to type VI collagen is lost in osteoarthritis.
NG2/human melanoma proteoglycan (HMPG) is a chondroitin sulphate proteoglycan (CSPG), expressed by chondrocytes in fetal and in normal and osteoarthritic (OA) adult articular cartilage. NG2/HMPG is a receptor for extracellular matrix proteins, including type VI collagen, and regulates beta1 integrin binding to fibronectin. This study was undertaken to identify whether NG2/HMPG had similar activities in human articular chondrocytes (HACs). Normal and OA adult HAC adhesion to fibronectin, type II or type VI collagen was assessed using a methylene blue assay. The requirement for integrins, NG2/HMPG, and integrin-associated signalling molecules was investigated using anti-beta1 integrin and anti-HMPG antibodies and pharmacological inhibitors of signalling molecules. The adhesion of normal and OA HACs to fibronectin, type II and type VI collagen was beta1 integrin-dependent. Normal HAC adhesion to type VI collagen was stimulated by anti-HMPG antibodies. This effect was inhibited by pertussis toxin. Anti-HMPG antibodies had no effect on OA chondrocyte adhesion to type VI collagen, or on normal and OA cell adhesion to fibronectin and type II collagen. The results show that NG2/HMPG modulates integrin-mediated interactions of normal HACs with type VI collagen. Loss of this activity may be of importance in the progression of osteoarthritis.


Matrix metalloproteinase-based photodynamic molecular beacons for targeted destruction of bone metastases in vivo.
The metastatic spread of cancer from the primary site or organ is one of its most devastating aspects, being responsible for up to 90% of cancer-associated mortality. Bone is one of the common sites of metastatic spread, including the vertebrae. Regardless of the treatment strategy, the clinical goals for patients with vertebral metastases are to improve the quality of life by preventing neurologic decline, to achieve durable pain relief and enhance local tumor control. However, in part due to the close proximity of the spinal cord, current treatment options are limited. We propose a novel therapeutic strategy with the use of photodynamic molecular beacons (PMBs) for targeted destruction of spinal metastases, particularly to de-bulk lesions as an adjuvant to vertebroplasty or kyphoplasty in order to mechanically stabilize weak or fractured vertebrae. The PDT efficacy of a matrix metalloproteinase-specific PMB is reported in a metstatic model that recapitulates the clinical features of tumor growth within the bone. We demonstrate that not only does tumor cell destruction occur but also the killing of bone stromal cells. The potential of PMB-PDT to destroy metastatic tumors, disrupt the osteolytic cycle and better preserve critical organs with an increased therapeutic window compared with conventional photosensitizers is demonstrated.


Current practice of difficult airway management: A survey.
Difficult airway (DA) management depends on both training and actual usage of the various approaches in the event of difficulty. The aim of the study was to assess how well the anaesthesiologists are equipped to deal with DA situations. The current practice preference of DA management was also assessed. A questionnaire was distributed in a continuing medical education (CME) programme dedicated to DA and responses were noted and analysed, using Statistical Package for Social Sciences (SPSS) version 18. The response rate was 73%. Airway assessment was performed by majority. Sixty eight percent consultants and 47% residents were well aware of the American Society of Anesthesiologists' DA algorithm. 67% consultants and 65% residents attended at least one CME on DA in the previous 5 years. There was an overall deficiency of video laryngoscopes, retrograde intubation and cricothyrotomy sets. Most of the respondents were comfortable in using supraglottic airway devices (SGADs). In anticipated DA, the preferred choice of management for junior doctors was attempting conventional method once and awake fibreoptic bronchoscopy (FOB) for the experienced. In unanticipated DA, most of the residents and consultants opted for SGAD. Extubation strategy was similar for both. Thirty four percent of respondents experienced a 'cannot intubate-cannot ventilate' situation at least once. Our survey showed that most respondents performed routine pre-operative airway assessment. A good armamentarium of airway gadgets should be made available in hospitals. Further training in techniques like video laryngoscopy, FOB or cricothyrotomy are essential.


Denitrifiers, nitrogen-fixing bacteria and N2O soil gas flux in high Arctic ice-wedge polygon cryosols.
Climate warming and subsequent permafrost thaw may result in organic carbon and nutrient stores being metabolized by microbial communities, resulting in a positive feedback loop of greenhouse gas (GHG) soil emissions. As the third most important GHG, understanding nitrous oxide (N2O) flux in Arctic mineral ice-wedge polygon cryosols and its relationship to the active microbial community is potentially a key parameter for understanding future GHG emissions and climatic warming potential. In the present study, metatranscriptomic analyses of active layer Arctic cryosols, at a representative ice-wedge polygon site, identified active nitrogen-fixing and denitrifying bacteria that included members of Rhizobiaceae, Nostocaceae, Cyanothecaceae, Rhodobacteraceae, Burkholderiaceae, Chloroflexaceae, Azotobacteraceae and Ectothiorhodospiraceae. Unique microbial assemblages with higher proportion of Rhodobacteriales and Rhocyclales were identified by targeted functional gene sequencing at locations with higher (P = 0.053) N2O emissions in the wetter trough soils compared with the dryer polygon interior soils. This coincided with a higher relative abundance of the denitrification nirS gene and higher nitrate/nitrite concentrations in trough soils. The elevated N2O flux observed from wetter trough soils compared with drier polygon interior soils is concerning from a climate warming perspective, since the Arctic is predicted to become warmer and wetter.


The interleukin-1 receptor/Toll-like receptor superfamily: signal transduction during inflammation and host defense.
The signal transduction pathways activated by the proinflammatory cytokine interleukin-1 (IL-1) have been the focus of much attention because of the important role that IL-1 plays in inflammatory diseases. A number of proteins have been described that participate in the post-receptor activation of the transcription factor NF-kappaB and stress-activated protein kinases such as p38 mitogen-activated protein kinase (MAPK). It has also emerged that the type I IL-1 receptor (termed IL-1RI) is a member of an expanding receptor superfamily. These related receptors all have sequence similarity in their cytosolic regions. The family includes the Drosophila melanogaster protein Toll, the IL-18 receptor (IL-18R), and the Toll-like receptors TLR-2 and TLR-4, which bind molecules from Gram-positive and Gram-negative bacteria, respectively. Because of the similarity of IL-1RI to Toll, the conserved sequence in the cytosolic region of these proteins has been termed the Toll-IL-1 receptor (TIR) domain. The same proteins activated during signaling by IL-1RI also participate in signaling by IL-18R and TLR-4. The receptor superfamily is evolutionarily conserved; members occur in plants and insects and also function in host defense. The signaling proteins activated are also conserved across species. This receptor superfamily therefore represents an ancient signaling system that is a critical determinant of the innate immune and inflammatory responses.


Bias-dependent local structure of water molecules at a metallic interface.
Understanding the local structure of water at the interfaces of metallic electrodes is a key issue in aqueous-based electrochemistry. Nevertheless a realistic simulation of such a setup is challenging, particularly when the electrodes are maintained at different potentials. To correctly compute the effect of an external bias potential applied to truly semi-infinite surfaces, we combine Density Functional Theory (DFT) and Non-Equilibrium Green's Function (NEGF) methods. This framework allows for the out-of-equilibrium calculation of forces and dynamics, and directly correlates to the chemical potential of the electrodes, which is introduced experimentally. In this work, we apply this methodology to study the electronic properties and atomic forces of a water molecule at the interface of a gold surface. We find that the water molecule tends to align its dipole moment with the electric field, and it is either repelled or attracted to the metal depending on the sign and magnitude of the applied bias, in an asymmetric fashion.


Chaperoning 5S RNA assembly.
In eukaryotes, three of the four ribosomal RNAs (rRNAs)—the 5.8S, 18S, and 25S/28S rRNAs—are processed from a single pre-rRNA transcript and assembled into ribosomes. The fourth rRNA, the 5S rRNA, is transcribed by RNA polymerase III and is assembled into the 5S ribonucleoprotein particle (RNP), containing ribosomal proteins Rpl5/uL18 and Rpl11/uL5, prior to its incorporation into preribosomes. In mammals, the 5S RNP is also a central regulator of the homeostasis of the tumor suppressor p53. The nucleolar localization of the 5S RNP and its assembly into preribosomes are performed by a specialized complex composed of Rpf2 and Rrs1 in yeast or Bxdc1 and hRrs1 in humans. Here we report the structural and functional characterization of the Rpf2-Rrs1 complex alone, in complex with the 5S RNA, and within pre-60S ribosomes. We show that the Rpf2-Rrs1 complex contains a specialized 5S RNA E-loop-binding module, contacts the Rpl5 protein, and also contacts the ribosome assembly factor Rsa4 and the 25S RNA. We propose that the Rpf2-Rrs1 complex establishes a network of interactions that guide the incorporation of the 5S RNP in preribosomes in the initial conformation prior to its rotation to form the central protuberance found in the mature large ribosomal subunit.


New Zealand's new alcohol laws: protocol for a mixed-methods evaluation.
Alcohol consumption is a major cause of mortality and morbidity globally. In response to strong calls from the public for alcohol law reform, the New Zealand Government recently reduced the blood alcohol limit for driving and introduced the Sale and Supply of Alcohol Act which aim to (1) improve community input into local decision-making on alcohol; (2) reduce the availability of alcohol; and (3) reduce hazardous drinking and alcohol-related harm. In this project we seek to evaluate the new laws in terms of these objectives. A policy evaluation framework is proposed to investigate the implementation and outcomes of the reforms. We will use quantitative and qualitative methods, employing a pre-post design. Participants include members of the public, local government staff, iwi (Māori tribal groups that function collectively to support their members) and community group representatives. Data will be collected via postal surveys, interviews and analysis of local government documents. Liquor licensing, police and hospital injury data will also be used. Community input into local government decision-making will be operationalised as: the number of objections per license application and the number of local governments adopting a local alcohol policy (LAP). Outcome measures will be the 'restrictiveness' of LAPs compared to previous policies, the number (per 1000 residents) and density (per square kilometre) of alcohol outlets throughout NZ, and the number of weekend late-night (i.e., post 10 pm) trading hours. For consumption and harm, outcomes will be the prevalence of hazardous drinking, harm from own and others' drinking, community amenity effects, rates of assault, and rates of alcohol-involved traffic crashes. Multiple regression will be used to model how the outcomes vary by local government area from before to after the law changes take effect. These measures will be complemented by qualitative analysis of LAP development and public participation in local decision-making on alcohol. The project will evaluate how well the reforms meet their explicit public health objectives.


Validating the Questionnaire for Verifying Stroke-Free Status (QVSFS) by neurological history and examination.
The Questionnaire for Verifying Stroke-Free Status (QVSFS) is an 8-item structured interview designed to identify stroke-free individuals. Previously, the QVSFS was validated with medical record review in a cohort with a low prevalence (7.1%) of stroke or transient ischemic attack (TIA). The objective of this study was to evaluate the validity of the QVSFS by comparing it with stroke status as determined by neurological history and examination in a population with a higher prevalence of stroke. A research assistant administered the QVSFS to outpatients from Veterans Administration stroke and general medicine clinics. Subjects were defined as QVSFS negative if responses to all 8 questions were negative. Questions requiring rephrasing or clarification were noted. Neurologists, blinded to QVSFS scores, interviewed and examined all subjects to determine stroke-free status, defined as no history or examination findings of previous stroke and/or TIA. One hundred fifty-five subjects were examined; mean age was 70 years; 98.1% were male. Seventy-eight subjects were determined to be stroke free by the neurologist. The negative predictive value of the QVSFS was 0.96, with positive predictive value of 0.71. No question required rephrasing or clarification >5 times. Twenty-two subjects (14.2%) required rephrasing or clarification of at least 1 question. The QVSFS can effectively identify stroke-free individuals with a high degree of accuracy, even in a population with a large proportion of patients with prior stroke or TIA. Accuracy for identifying subjects with stroke and/or TIA is lower, but the QVSFS may still be useful as a screening tool in that regard.


Structures of normal single-stranded DNA and deoxyribo-3'-S-phosphorothiolates bound to the 3'-5' exonucleolytic active site of DNA polymerase I from Escherichia coli.
The interaction of a divalent metal ion with a leaving 3' oxygen is a central component of several proposed mechanisms of phosphoryl transfer. In support of this are recent kinetic studies showing that thiophilic metal ions (e.g., Mn2+) stimulate the hydrolysis of compounds in which sulfur takes the place of the leaving oxygen. To examine the structural basis of this phenomenon, we have solved four crystal structures of single-stranded DNA's containing either oxygen or sulfur at a 3'-bridging position bound in conjunction with various metal ions at the 3'-5' exonucleolytic active site of the Klenow fragment (KF) of DNA polymerase I from Escherichia coli. Two structures of normal ssDNA bound to KF in the presence of Zn2+ and Mn2+ or Zn2+ alone were refined at 2.6- and 2.25-A resolution, respectively. They serve as standards for comparison with other Mn2+- and Zn2+-containing structures. In these cases, Mn2+ and Zn2+ bind at metal ion site B in a nearly identical position to Mg2+ (Brautigam and Steitz (1998) J. Mol. Biol. 277, 363-377). Two structures of KF bound to a deoxyoligonucleotide that contained a 3'-bridging sulfur at the scissile phosphate were refined at 2.03-A resolution. Although the bridging sulfur compounds bind in a manner very similar to that of the normal oligonucleotides, the presence of the sulfur changes the metal ion binding properties of the active site such that Mn2+ and Zn2+ are observed at metal ion site B, but Mg2+ is not. It therefore appears that the ability of the bridging sulfur compounds to exclude nonthiophilic metal ions from metal ion site B explains the low activity of KF exonuclease on these substrates in the presence of Mg2+ (Curley et al. (1997) J. Am. Chem. Soc. 119, 12691-12692) and that the 3'-bridging atom of the substrate is influencing the binding of metal ion B prior to catalysis.


Live-Cell RNA Imaging with Metabolically Incorporated Fluorescent Nucleosides.
Fluorescence imaging is a powerful method for probing macromolecular dynamics in biological systems; however, approaches for cellular RNA imaging are limited to the investigation of individual RNA constructs or bulk RNA labeling methods compatible primarily with fixed samples. Here, we develop a platform for fluorescence imaging of bulk RNA dynamics in living cells. We show that fluorescent bicyclic and tricyclic cytidine analogues can be metabolically incorporated into cellular RNA by overexpression of uridine-cytidine kinase 2. In particular, metabolic feeding with the tricyclic cytidine-derived nucleoside tC combined with confocal imaging enables the investigation of RNA synthesis, degradation, and trafficking at single-cell resolution. We apply our imaging modality to study RNA metabolism and localization during the oxidative stress response and find that bulk RNA turnover is greatly accelerated upon NaAsO2 treatment. Furthermore, we identify cytoplasmic RNA granules containing RNA transcripts generated during oxidative stress that are distinct from canonical stress granules and P-bodies and co-localize with the RNA helicase DDX6. Taken together, our work provides a powerful approach for live-cell RNA imaging and reveals how cells reshape RNA transcriptome dynamics in response to oxidative stress.


Perspectives on the Use of Ninjin'yoeito in Modern Medicine: A Review of Randomized Controlled Trials.
Ninjin'yoeito (NYT), a traditional Japanese (Kampo) medicine that originates from China, has been used to treat qi and blood deficiency based on its original concept. Kampo medicine has been widely used to treat many conditions and disorders combined with western medicine or Kampo medicine alone in modern situation. We reviewed randomized controlled trials (RCTs) of NYT and discussed various standpoints regarding its use in modern situation. We searched PubMed, Cochrane Library, and Evidence Reports of Kampo Treatment (EKAT) for articles written in English, and Ichushi, J-Stage, and EKAT for those written in Japanese. Articles published before January 1, 2019, were retrieved using the keywords "ninjinyoeito" and "ninjin'yoeito," and RCTs were selected from these extracted articles. Of 734 articles, 13 were RCTs, 46 were non-RCTs or studies of other designs, 15 were case reports, and 36 were experimental studies using NYT. NYT was evaluated for its use as a treatment for cancer and related conditions, refractory blood diseases and conditions, and otorhinolaryngologic symptom in 13 RCTs. Based on the use of Kampo medicine in modern situation called as Yasui's classification, 10 of 13 RCTs were categorized as "the side effects of Western medicinal treatment are mitigated when combined with Kampo treatment" and the remaining 3 were categorized as "treatment effect of Kampo medicine is increased in combination with standard Western medicinal treatment." Conclusion. Several studies demonstrated the efficacy of NYT in refractory diseases and other conditions, and the accompanied side effects of treatment with western medicine.


Crowding increases salivary cortisol but not self-directed behavior in captive baboons.
Reduced space can lead to crowding in social animals. Crowding increases the risk of agonistic interactions that, in turn, may require additional physiological defensive coping mechanisms affecting health. To determine the stress induced from increased social density in a group of nineteen baboons living in an indoor/outdoor enclosure, saliva cortisol levels and rates of anxiety-related behavior were analyzed across two unique crowding episodes. Initially, mean salivary cortisol levels when animals were restricted to their indoor quarters were compared to those when they also had access to their larger outdoor enclosure. Then, mean cortisol levels were compared before, during, and after two distinct crowding periods of long and short duration. Crowding resulted in significantly elevated cortisol during crowding periods compared to non-crowded periods. Cortisol levels returned to baseline following two crowding episodes contrasting in their length and ambient climate conditions. These cortisol elevations indicate greater metabolic costs of maintaining homeostasis under social stress resulting from reduced space. Self-directed behavior, conversely, was not reliably elevated during crowding. Results suggest that the potential for negative social interactions, and/or the uncertainty associated with social threat can cause physiological stress responses detected by salivary cortisol. Self-directed behavioral measures of stress may constitute inadequate indicators of social stress in colony-housed monkeys or represent subjective emotional arousal unrelated to hypothalamic-pituitary adrenal axis activation.


Designing novel hybrid materials by one-pot co-condensation: from hydrophobic mesoporous silica nanoparticles to superamphiphobic cotton textiles.
This work reports the synthesis and characterization of mesoporous silica nanoparticles (MSNs) functionalized with tridecafluorooctyltriethoxysilane (F13) and their in situ incorporation onto cotton textiles. The hybrid MSNs and the functional textiles were prepared by a one-pot co-condensation methodology between tetraethylorthosilicate (TEOS) and F13, with hexadecyltrimethylammonium chloride (CTAC) as the template and triethanolamine as the base. The influence of the F13 to TEOS molar ratio (1:10, 1:5 and 1:3) on the nanoparticle morphology, porosity, degree of functionalization, and hydro/oleophobic properties is discussed. The hybrid nanosilicas presented high colloidal stability and were spherical and monodispersed with average particle size of ∼45 nm. They also showed high surface areas, large pore volumes, and a wormhole-type mesoporous structure. The increase in the organosilane proportion during the co-condensation process led to a more radially branched wormhole-like mesoporosity, a decrease in the surface area, pore volume, and amount of surface silanol groups, and an enrichment of the surface with fluorocarbon moieties. These changes imparted hydrophobic and oleophobic properties to the materials, especially to that containing the highest F13 loading. Cotton textiles were coated with the F13-MSNs through an efficient and less time-consuming route. The combination between surface roughness and mesoporosity imparted by the MSNs, and the low surface energy provided by the organosilane resulted in superhydrophobic functional textiles. Moreover, the textile with the highest loading of fluorocarbon groups was superamphiphobic.


Reliability of causality assessment for drug, herbal and dietary supplement hepatotoxicity in the Drug-Induced Liver Injury Network (DILIN).
Because of the lack of objective tests to diagnose drug-induced liver injury (DILI), causality assessment is a matter of debate. Expert opinion is often used in research and industry, but its test-retest reliability is unknown. To determine the test-retest reliability of the expert opinion process used by the Drug-Induced Liver Injury Network (DILIN). Three DILIN hepatologists adjudicate suspected hepatotoxicity cases to one of five categories representing levels of likelihood of DILI. Adjudication is based on retrospective assessment of gathered case data that include prospective follow-up information. One hundred randomly selected DILIN cases were re-assessed using the same processes for initial assessment but by three different reviewers in 92% of cases. The median time between assessments was 938 days (range 140-2352). Thirty-one cases involved >1 agent. Weighted kappa statistics for overall case and individual agent category agreement were 0.60 (95% CI: 0.50-0.71) and 0.60 (0.52-0.68) respectively. Overall case adjudications were within one category of each other 93% of the time, while 5% differed by two categories and 2% differed by three categories. Fourteen per cent crossed the 50% threshold of likelihood owing to competing diagnoses or atypical timing between drug exposure and injury. The DILIN expert opinion causality assessment method has moderate interobserver reliability but very good agreement within one category. A small but important proportion of cases could not be reliably diagnosed as ≥50% likely to be DILI.


A network model for the specification of vulval precursor cells and cell fusion control in Caenorhabditis elegans.
The vulva of Caenorhabditis elegans has been long used as an experimental model of cell differentiation and organogenesis. While it is known that the signaling cascades of Wnt, Ras/MAPK, and NOTCH interact to form a molecular network, there is no consensus regarding its precise topology and dynamical properties. We inferred the molecular network, and developed a multivalued synchronous discrete dynamic model to study its behavior. The model reproduces the patterns of activation reported for the following types of cell: vulval precursor, first fate, second fate, second fate with reversed polarity, third fate, and fusion fate. We simulated the fusion of cells, the determination of the first, second, and third fates, as well as the transition from the second to the first fate. We also used the model to simulate all possible single loss- and gain-of-function mutants, as well as some relevant double and triple mutants. Importantly, we associated most of these simulated mutants to multivulva, vulvaless, egg-laying defective, or defective polarity phenotypes. The model shows that it is necessary for RAL-1 to activate NOTCH signaling, since the repression of LIN-45 by RAL-1 would not suffice for a proper second fate determination in an environment lacking DSL ligands. We also found that the model requires the complex formed by LAG-1, LIN-12, and SEL-8 to inhibit the transcription of eff-1 in second fate cells. Our model is the largest reconstruction to date of the molecular network controlling the specification of vulval precursor cells and cell fusion control in C. elegans. According to our model, the process of fate determination in the vulval precursor cells is reversible, at least until either the cells fuse with the ventral hypoderm or divide, and therefore the cell fates must be maintained by the presence of extracellular signals.


Physical activity and sedentary activity patterns among children and adolescents: a latent class analysis approach.
While much is known about the overall levels of physical activity and sedentary activity among youth, few studies have attempted to define clusters of such behaviors. The purpose of this study was to identify and describe unique classes of youth based on their participation in a variety of physical activity and sedentary behaviors. Latent class analysis was used to characterize segments of youth based on patterns of self-reported and accelerometer-measured participation in 12 behaviors. Children and adolescents (N=720) from 6th-11th grade were included in the analysis. Differences in class membership were examined using multinomial logistic regression. Three distinct classes emerged for boys and girls. Among boys, the 3 classes were characterized as "Active" (42.1%), "Sedentary" (24.9%), and "Low Media/Moderate Activity" (33.0%). For girls, classes were "Active" (18.7%), "Sedentary" (47.6%), and "Low Media/Functional Activity" (33.7%). Significant differences were found between the classes for a number of demographic indicators including the proportion in each class who were classified as overweight or obese. The behavioral profiles of the classes identified in this study can be used to suggest possible audience segments for intervention and to tailor strategies appropriately.


The relationship between individual empowerment and health-promoting lifestyle among women NGOs in northern Iran.
According to the health-promoting approach, people should be empowered such that they take responsibility for their health and follow a healthy lifestyle. Empowerment is a process in which people confront problems and tasks in their lives in order to better control them. This study was conducted to specify the relationship between individual empowerment and health-promoting lifestyle among women NGOs of northern Iran. In this cross-sectional study, 290 women NGOs of Guilan Province were selected randomly using multistage cluster sampling, and were examined using the questionnaire of health-promoting lifestyle profile II and individual empowerment inventory scale. Data were analyzed using STATA 11 software via one-way ANOVA, Pearson correlation coefficient and multivariate linear regression. Both variables of individual empowerment and health-promoting lifestyle were of favorable status among the population under study. The highest score in individual empowerment belonged to the domain of social support and the highest score in lifestyle belonged to spiritual growth. A significant relationship was found between individual empowerment and health-promoting lifestyle (p<0.001). The highest correlation between individual empowerment and dimensions of health-promoting lifestyle was related to interpersonal relationships. In addition, it was specified that 21% of lifestyle variance could be explained by individual empowerment. There is a relation between individual empowerment and health-promoting lifestyle. Individual empowerment is a predictive variable to have a health-promoting lifestyle. Therefore, by increasing individuals' empowerment, their healthy lifestyle can be promoted.


Distribution of diatoms in relation to land use and pH in blackwater coastal plain streams.
We compared the composition of diatom assemblages collected from New Jersey Pinelands blackwater streams draining four different land uses, including forest land, abandoned-cranberry bogs, active-cranberry bogs, and developed and upland-agricultural land. Over a 2-year period (2002-2003), we collected 132 diatom taxa at 14 stream sites. Between-year variability in the composition of stream samples was high. Most diatom species were rarely encountered and were found in low abundance. Specific conductance and pH were higher at developed/agricultural sites compared with all other site types. Neither species richness nor genus richness was significantly different between stream types. However, clear community patterns were evident, and a significant difference in species composition existed between the developed/agricultural sites and both cranberry and forest sites. The primary community gradient, represented by the first axis of a DCA ordination, was associated with variations in pH and specific conductance. Although community patterns revealed by ordinating the data collected in 2002 differed from those obtained using the 2003 data, both ordinations contrasted the developed/agricultural sites and the other sites. Acidobiontic and acidophilous diatoms characterized the dominant species at forest, abandoned-bog, and cranberry sites, whereas indifferent species dominated the developed/agricultural samples. Although our study demonstrated a relationship between the composition of diatom assemblages and watershed conditions, several factors, including taxonomic problems, the large number of diatom species, incomplete pH classifications, and year-to-year variability may limit the utility of diatom species as indicators of watershed conditions in the New Jersey Pinelands.


Organ-specific radiation dose rates and effective dose rates during percutaneous nephrolithotomy.
Radiation exposure during medical procedures continues to be an increasing concern for physicians and patients. We determined organ-specific dose rates and calculated effective dose rates during right and left percutaneous nephrolithotomy (PCNL) using a validated phantom model. A validated anthropomorphic adult male phantom was placed prone on an operating room table. Metal oxide semiconductor field effect transistor dosimeters were placed at 20 organ locations in the model and were used to measure the organ dosages. A portable C-arm was used to provide continuous fluoroscopy for three 10 minute runs each to simulate a left and right PCNL. Organ dose rate (mGy/s) was determined by dividing organ dose by fluoroscopy time. The organ dose rates were multiplied by their tissue weighting factor and summed to determine effective dose rate (EDR) (mSv/s). Two-dimensional radiation distribution in the abdomen during a left-sided PCNL was visually determined using radiochromic film. The EDR for a left PCNL was 0.021 mSv/s ± 0.0008. The EDR for a right PCNL was 0.014 mSv/s ± 0.0004. The skin entrance was exposed to the greatest amount of radiation during left and right PCNL, 0.24 mGy/s and 0.26 mGy/s, respectively. Radiochromic film demonstrates visually the nonuniform dose distribution as the x-ray beam enters through the skin from the radiation source. The effective dose rate is higher for a left-sided PCNL compared with a right-sided PCNL. The distribution of radiation exposure during PCNL is not uniform. Further studies are needed to determine the long-term implications of these radiation doses during percutaneous stone removal.


A genome-scale RNA-interference screen identifies RRAS signaling as a pathologic feature of Huntington's disease.
A genome-scale RNAi screen was performed in a mammalian cell-based assay to identify modifiers of mutant huntingtin toxicity. Ontology analysis of suppressor data identified processes previously implicated in Huntington's disease, including proteolysis, glutamate excitotoxicity, and mitochondrial dysfunction. In addition to established mechanisms, the screen identified multiple components of the RRAS signaling pathway as loss-of-function suppressors of mutant huntingtin toxicity in human and mouse cell models. Loss-of-function in orthologous RRAS pathway members also suppressed motor dysfunction in a Drosophila model of Huntington's disease. Abnormal activation of RRAS and a down-stream effector, RAF1, was observed in cellular models and a mouse model of Huntington's disease. We also observe co-localization of RRAS and mutant huntingtin in cells and in mouse striatum, suggesting that activation of R-Ras may occur through protein interaction. These data indicate that mutant huntingtin exerts a pathogenic effect on this pathway that can be corrected at multiple intervention points including RRAS, FNTA/B, PIN1, and PLK1. Consistent with these results, chemical inhibition of farnesyltransferase can also suppress mutant huntingtin toxicity. These data suggest that pharmacological inhibition of RRAS signaling may confer therapeutic benefit in Huntington's disease.


Socioeconomic inequalities in cause-specific mortality in 15 European cities.
Socioeconomic inequalities are increasingly recognised as an important public health issue, although their role in the leading causes of mortality in urban areas in Europe has not been fully evaluated. In this study, we used data from the INEQ-CITIES study to analyse inequalities in cause-specific mortality in 15 European cities at the beginning of the 21st century. A cross-sectional ecological study was carried out to analyse 9 of the leading specific causes of death in small areas from 15 European cities. Using a hierarchical Bayesian spatial model, we estimated smoothed Standardized Mortality Ratios, relative risks and 95% credible intervals for cause-specific mortality in relation to a socioeconomic deprivation index, separately for men and women. We detected spatial socioeconomic inequalities for most causes of mortality studied, although these inequalities differed markedly between cities, being more pronounced in Northern and Central-Eastern Europe. In the majority of cities, most of these causes of death were positively associated with deprivation among men, with the exception of prostatic cancer. Among women, diabetes, ischaemic heart disease, chronic liver diseases and respiratory diseases were also positively associated with deprivation in most cities. Lung cancer mortality was positively associated with deprivation in Northern European cities and in Kosice, but this association was non-existent or even negative in Southern European cities. Finally, breast cancer risk was inversely associated with deprivation in three Southern European cities. The results confirm the existence of socioeconomic inequalities in many of the main causes of mortality, and reveal variations in their magnitude between different European cities.


Biogeography and evolution of the Carassius auratus-complex in East Asia.
Carassius auratus is a primary freshwater fish with bisexual diploid and unisexual gynogenetic triploid lineages. It is distributed widely in Eurasia and is especially common in East Asia. Although several genetic studies have been conducted on C. auratus, they have not provided clear phylogenetic and evolutionary descriptions of this fish, probably due to selection bias in sampling sites and the DNA regions analysed. As the first step in clarifying the evolutionary entity of the world's Carassius fishes, we attempted to clarify the phylogeny of C. auratus populations distributed in East Asia. We conducted a detailed analysis of a large dataset of mitochondrial gene sequences [CR, 323 bp, 672 sequences (528 sequenced + 144 downloaded); CR + ND4 + ND5 + cyt b, 4669 bp in total, 53 sequences] obtained from C. auratus in East Asia. Our phylogeographic analysis revealed two superlineages, one distributed mainly among the Japanese main islands and the other in various regions in and around the Eurasian continent, including the Ryukyus and Taiwan. The two superlineages include seven lineages with high regional specificity that are composed of endemic populations indigenous to each region. The divergence time of the seven lineages was estimated to be 0.2 million years ago (Mya) by a fossil-based method and 1.0-1.9 Mya by the molecular clock method. The antiquity and endemism of these lineages suggest that they are native to their respective regions, although some seem to have been affected by the artificial introduction of C. auratus belonging to other lineages. Triploids of C. auratus did not form a monophyletic lineage but were clustered mostly with sympatric diploids. The results of the present study revealed the existence of two superlineages of C. auratus in East Asia that include seven lineages endemic to each of the seven regions examined. The lack of substantial genetic separation between triploids and diploids indicates that triploids are not composed of a single independent lineage. The ancient origins and evolutionary uniqueness of the seven lineages warrant their conservation. An overall phylogenetic framework obtained from the present study will be of use for estimating the phylogenetic relationships of Carassius fishes on the Eurasian continent.


Optimal lesion assessment following acute radio frequency ablation of porcine kidney: cellular viability or histopathology?
Radio frequency ablation (RFA) has been used as a minimally invasive alternative to nephrectomy for small renal tumors. Questions have arisen regarding the accuracy of cell viability determination on standard hematoxylin and eosin (H & E) staining. We investigated and compared the histological characteristics of RF ablated renal tissue using nicotinamide adenine dinucleotide (NADH) and H & E staining. Ten porcine kidneys underwent laparoscopic RFA of the upper and lower poles using a 2 (8) or 3 cm (2) protocol with 2 cycles of 90 W, target temperature 105C and treatment time 5.5 minutes per cycle. Following tract ablation the kidneys were immediately harvested, gross lesion size was measured and tissue was processed for standard H & E and NADH staining. H & E staining of ablated tissue revealed a number of alterations in renal tubular histology. However, all of these findings were focal with areas of parenchyma that appeared well preserved. Corresponding areas on NADH processed sections showed the complete absence of staining, indicating the lack of cellular viability. There were no skip areas noted on NADH processed sections and treated portions demonstrated a well demarcated border of ablation. While RFA produces discernible histological changes acutely on H & E, these alterations are variable and patchy, and they alternate with areas of well preserved tissue. Therefore, NADH staining should always be used to assess and verify cellular death in RFA lesions. In this study no skip areas of viable cells were noted within areas of ablated tissue on NADH staining.


A Shigella effector dampens inflammation by regulating epithelial release of danger signal ATP through production of the lipid mediator PtdIns5P.
Upon infection with Shigella flexneri, epithelial cells release ATP through connexin hemichannels. However, the pathophysiological consequence and the regulation of this process are unclear. Here we showed that in intestinal epithelial cell ATP release was an early alert response to infection with enteric pathogens that eventually promoted inflammation of the gut. Shigella evolved to escape this inflammatory reaction by its type III secretion effector IpgD, which blocked hemichannels via the production of the lipid PtdIns5P. Infection with an ipgD mutant resulted in rapid hemichannel-dependent accumulation of extracellular ATP in vitro and in vivo, which preceded the onset of inflammation. At later stages of infection, ipgD-deficient Shigella caused strong intestinal inflammation owing to extracellular ATP. We therefore describe a new paradigm of host-pathogen interaction based on endogenous danger signaling and identify extracellular ATP as key regulator of mucosal inflammation during infection. Our data provide new angles of attack for the development of anti-inflammatory molecules.


Unusual meningitis caused by non-typhoid Salmonella in an Italian infant: a case report.
Non-typhoid Salmonella (NTS) is an important cause of bacterial meningitis in newborn and infants in developing countries, but rarely in industrialized ones. We describe an unusual presentation of bacterial meningitis in an infant, focusing on his diagnostic and therapeutic management. An Italian two-month old male presented high fever and diarrhea with blood, associated with irritability. Inflammatory markers were high, cerebrospinal fluid analysis was compatible with bacterial meningitides but microbiological investigations were negative. Salmonella enteritidis was isolated from blood. Cerebral ultrasound and MRI showed periencephalic collection of purulent material. Specific antibiotic therapy with cefotaxime was initiated with improvement of clinical conditions and blood tests. Brain MRI follow up improved progressively. Most of pediatric patients with NTS infection develop self-limited gastroenteritis, but in 3-8% of the cases complications such as bacteremia and meningitis may occur, especially in weak patients. Cerebral imaging can be useful to identify neurological findings. Although there is no standardized treatment for this condition, specific antibiotic therapy for at least four weeks is recommended. Neuroimaging follow up is required due to high risk of relapse.