Cancer prevalence in Italian regions with local cancer registries.
To provide estimates and projections of cancer incidence and prevalence for those Italian regions whose population is partially covered by a cancer registry (CR) and to determine to what extent local CRs can be considered representative of the region, thus improving the potential of the information provided by CRs. A statistical method, MIAMOD (mortality-incidence analysis model), was used to estimate regional cancer incidence and prevalence from regional cancer mortality data and patient survival data recorded by the cancer registries. Estimates of the cancer incidence and prevalence in the various regions have thus been obtained for a number of major cancer sitas. A first and important step in validating the regional estimates has been the comparison of the MIAMOD estimates in the areas covered by the cancer registries with empirical incidence and prevalence observed by CRs, in order to assess the consistency in data, methods and assumptions. Empirical prevalence has been calculated by counting patients with a diagnosis of cancer who were alive on the reference date by PREVAL method. A correction factor has been applied to include patients diagnosed before the period of activity of the registry. General consistency was found between empirical and estimated (by MIAMOD) incidence and prevalence in the registry areas, which is indicative of the quality and the completeness of all data involved as well as the appropriateness of model choices. The prevalence of all cancers combined for Italian regions with CRs was estimated and projected to the year 2000 as ranging between 1,240 per 100,000 in Sicilia and 2,781 in Emilia-Romagna for men, while for women these figures were 1,765 in Sicilia and 4,019 in Liguria. Comparison of cancer prevalence in CR areas with regional estimates shows quite good consistency for Piemonte, Liguria and Lombardia, which means that the local CRs (of Torino, Genova and Varese, respectively) are representative of their respective regions. Prevalence in Emilia-Romagna appears to be rather well represented by only one, the Parma CR, of the three local CRs. The southern Italian registries of Latina and Ragusa recorded a lower cancer prevalence than was actually estimated in their respective regions. Cancer registries with a longer period of activity showed better agreement between empirical and estimated figures due to the more precise information provided, particularly regarding survival and incidence trends. In conclusion, this work shows the potential of the cancer registries not only to represent their population with respect to cancer morbidity but also as an invaluable tool to extrapolate this information to the larger areas they represent.


Secondary Use of Electronic Health Record Data for Prediction of Outpatient Visit Length in Ophthalmology Clinics.
Electronic health record systems have dramatically transformed the process of medical care, but one challenge has been increased time requirements for physicians. In this study, we address this challenge by developing and validating analytic models for predicting patient encounter length based on secondary EHR data. Key findings from this study are: (1) Secondary use of EHR data may be captured to predict provider interaction time with patients; (2) Modeling results using secondary data may provide more accurate predictions of provider interaction time than an expert provide; (3) These findings suggest that secondary use of EHR data may be used to develop effective customized scheduling methods to improve clinical efficiency. In the future, this has the potential to contribute toward methods for improved clinical scheduling and efficiency.


Initial human experience with Rubidium-82 renal PET/CT imaging.
Preclinical data have shown that Rubidium-82 chloride ((82)Rb) is a radiotracer with high first pass extraction and slow washout in the kidneys. The goal of this study was to investigate the feasibility of human kidney imaging with (82)Rb positron emission tomography (PET) and obtain quantitative data of its uptake non-invasively. Eight healthy volunteers underwent dynamic PET/CT imaging with (82)Rb. A preprogrammed pump was used to insure reproducible injections. Tissue time activity curves were generated from the renal cortex. An input function was derived from the left ventricular blood pool (LVBP), the descending thoracic aorta and the abdominal aorta. Renal blood flow was estimated by applying a two-compartment kinetic model. Results obtained with different input functions were compared. Radiotracer accumulation was rapid and reached a plateau within 15-30 s after the bolus entered the kidneys. The derived K1 and k2 parameters were reproducible using input functions obtained from diverse vascular locations. K1 averaged 1.98 ± 0.14 mL/min/g. The average k2 was 0.35 ± 0.11/min. Correlation between K1 values obtained from the LVBP from different bed positions when the kidneys and abdominal aorta were in the same field of view was excellent (R = 0.95). Non-invasive quantitative human kidney imaging with (82)Rb PET is feasible. Advantages of renal PET with (82)Rb include excellent image quality with high image resolution and contrast. (82)Rb has potential as a clinical renal imaging agent in humans.


First Trimester Gaze Pattern Estimation Using Stochastic Augmentation Policy Search for Single Frame Saliency Prediction.
While performing an ultrasound (US) scan, sonographers direct their gaze at regions of interest to verify that the correct plane is acquired and to interpret the acquisition frame. Predicting sonographer gaze on US videos is useful for identification of spatio-temporal patterns that are important for US scanning. This paper investigates utilizing sonographer gaze, in the form of gaze-tracking data, in a multimodal imaging deep learning framework to assist the analysis of the first trimester fetal ultrasound scan. Specifically, we propose an encoderdecoder convolutional neural network with skip connections to predict the visual gaze for each frame using 115 first trimester ultrasound videos; 29,250 video frames for training, 7,290 for validation and 9,126 for testing. We find that the dataset of our size benefits from automated data augmentation, which in turn, alleviates model overfitting and reduces structural variation imbalance of US anatomical views between the training and test datasets. Specifically, we employ a stochastic augmentation policy search method to improve segmentation performance. Using the learnt policies, our models outperform the baseline: KLD, SIM, NSS and CC (2.16, 0.27, 4.34 and 0.39 versus 3.17, 0.21, 2.92 and 0.28).


Time course of spatial contextual interference: event history analyses of simultaneous masking by nonoverlapping patterns.
Simultaneous masking refers to the impairment of performance on a visual target by simultaneously presented flankers. Whereas the spatial aspects of simultaneous masking have been studied extensively, the time course of these spatial influences is much less well understood. We here measure response latency and accuracy in a simultaneous masking paradigm and apply event history analysis to study the time course of target-flanker interactions. In our experiments, we presented a central target vernier flanked on both sides by 12 aligned distractor verniers that were either shorter, longer, or equal in length (Experiment 1), and that also were congruent or incongruent in their spatial offset with the target (Experiment 2). Response time distributions showed that there were more fast responses when the target was flanked by short flankers. Conditional accuracy functions showed that accuracy of responses dropped when the flankers had the same length as the target, but only for slow responses. These results are at odds with accounts based solely on lateral neural interactions or response competition, and instead suggest that top-down visual object-to-feature interference occurs when the target is not selected fast enough, congruent with object substitution theory.


Characterization of the annexin I gene and evaluation of its role in type 2 diabetes.
In a previous study, we identified suggestive linkage between type 2 diabetes and a locus on chromosome 9p13-q21. This region contains the gene annexin I (ANXA1), encoding a protein suggested to be involved in both insulin secretion and insulin action. In this study, we sequenced the exon/intron boundaries of the human ANXA1 gene and performed mutation screening in 41 individuals from the initial linkage study. We identified five single nucleotide polymorphisms A58G, A401G, intronic variance sequence (IVS)8-28A/G, IVS11 +31A/G, and IVS12-11T/G, which were further tested for association to diabetes in 197 parent/offspring trios using the transmission disequilibrium test. No significant association with type 2 diabetes was observed, although the common A allele of the +58A/G variant gave a 22:12 transmission distortion (P = 0.12). This variant was further genotyped in 481 case and control subjects, but no difference in allele, genotype, or haplotype frequencies were observed between the groups. Further, a novel polymorphic (CA)(15-25) repeat in intron 11 was genotyped in the subjects included in the initial linkage study. No improvement of the original finding was observed. We therefore concluded that the ANXA1 gene is unlikely to harbor variants that contribute to risk of type 2 diabetes.


Dosimetric comparison of standard three-dimensional conformal radiotherapy followed by intensity-modulated radiotherapy boost schedule (sequential IMRT plan) with simultaneous integrated boost-IMRT (SIB IMRT) treatment plan in patients with localized carcinoma prostate.
DOSIMETERIC AND RADIOBIOLOGICAL COMPARISON OF TWO RADIATION SCHEDULES IN LOCALIZED CARCINOMA PROSTATE: Standard Three-Dimensional Conformal Radiotherapy (3DCRT) followed by Intensity Modulated Radiotherapy (IMRT) boost (sequential-IMRT) with Simultaneous Integrated Boost IMRT (SIB-IMRT). Thirty patients were enrolled. In all, the target consisted of PTV P + SV (Prostate and seminal vesicles) and PTV LN (lymph nodes) where PTV refers to planning target volume and the critical structures included: bladder, rectum and small bowel. All patients were treated with sequential-IMRT plan, but for dosimetric comparison, SIB-IMRT plan was also created. The prescription dose to PTV P + SV was 74 Gy in both strategies but with different dose per fraction, however, the dose to PTV LN was 50 Gy delivered in 25 fractions over 5 weeks for sequential-IMRT and 54 Gy delivered in 27 fractions over 5.5 weeks for SIB-IMRT. The treatment plans were compared in terms of dose-volume histograms. Also, Tumor Control Probability (TCP) and Normal Tissue Complication Probability (NTCP) obtained with the two plans were compared. The volume of rectum receiving 70 Gy or more (V > 70 Gy) was reduced to 18.23% with SIB-IMRT from 22.81% with sequential-IMRT. SIB-IMRT reduced the mean doses to both bladder and rectum by 13% and 17%, respectively, as compared to sequential-IMRT. NTCP of 0.86 ± 0.75% and 0.01 ± 0.02% for the bladder, 5.87 ± 2.58% and 4.31 ± 2.61% for the rectum and 8.83 ± 7.08% and 8.25 ± 7.98% for the bowel was seen with sequential-IMRT and SIB-IMRT plans respectively. For equal PTV coverage, SIB-IMRT markedly reduced doses to critical structures, therefore should be considered as the strategy for dose escalation. SIB-IMRT achieves lesser NTCP than sequential-IMRT.


A multidisciplinary method to map potential tuberculosis transmission 'hot spots' in high-burden communities.
Global control of the tuberculosis (TB) epidemic remains poor, especially in high-burden settings where ongoing transmission sustains the epidemic. In such settings, a significant amount of transmission takes place outside the household, and practical approaches to understanding transmission at community level are needed. To identify and map potential TB transmission 'hot spots' across high-burden communities. Our method draws on data that qualitatively describe a high-burden community in Cape Town, South Africa. Established transmission principles are applied to grade the potential TB transmission risk posed by congregate settings in the community. Geographic information systems (GIS) technology then creates a visual map, locating potential transmission 'hot spots' in the community. Drinking places (shebeens), clinics and churches (often gatherings in confined homes) emerge as gathering places that potentially pose a high transmission risk, particularly if located in overcrowded and impoverished areas of the community. This proof-of-concept study demonstrates that combining qualitative techniques with GIS mapping may improve our understanding of potential TB transmission within a community and guide public health interventions to enhance TB control efforts.


Association of Longitudinal Trajectories of Systolic BP with Risk of Incident CKD: Results from the Korean Genome and Epidemiology Study.
Although hypertension is a well known risk factor for CKD, few studies have evaluated the association between temporal trends of systolic BP and kidney function decline in persons without hypertension. We studied whether changes in systolic BP over time could influence incident CKD development in 4643 individuals without CKD and hypertension participating in the Korean Genome and Epidemiology Study, a prospective community-based cohort study. Using group-based trajectory modeling, we categorized three distinct systolic BP trajectories: decreasing, stable, and increasing. The primary outcome was incident CKD development, defined as two consecutive eGFR measurements <60 ml/min per 1.73 m2. Among participants with an increasing systolic BP trajectory, systolic BP increased from 105 to 124 mm Hg. During 31,936 person-years of follow-up (median 7.7 years), 339 participants developed incident CKD. CKD incidence rates were 8.9, 9.6, and 17.8 cases per 1000 person-years in participants with decreasing, stable, and increasing systolic BP trajectories, respectively. In multivariable cause-specific Cox analysis, after adjustment of baseline eGFR, systolic BP, and other confounders, increasing systolic BP trajectory associated with a 1.57-fold higher risk of incident CKD (95% confidence interval, 1.20 to 2.06) compared with a stable trajectory. There was a significant effect modification of baseline systolic BP on the association between systolic BP trajectories and CKD risk (P value for interaction =0.02), and this association was particularly evident in participants with baseline systolic BP <120 mm Hg. In addition, increasing systolic BP trajectory versus a stable trajectory was associated with higher risk of new development of albuminuria. Increasing systolic BP over time without reaching the hypertension threshold is associated with a significantly increased risk of incident CKD in healthy adults.


A comparison of severe hemodynamic disturbances between dexmedetomidine and propofol for sedation in neurocritical care patients.
Dexmedetomidine and propofol are commonly used sedatives in neurocritical care as they allow for frequent neurologic examinations. However, both agents are associated with significant hemodynamic side effects. The primary objective of this study is to compare the prevalence of severe hemodynamic effects in neurocritical care patients receiving dexmedetomidine and propofol. Multicenter, retrospective, propensity-matched cohort study. Neurocritical care units at two academic medical centers with dedicated neurocritical care teams and board-certified neurointensivists. Neurocritical care patients admitted between July 2009 and September 2012 were evaluated and then matched 1:1 based on propensity scoring of baseline characteristics. Continuous sedation with dexmedetomidine or propofol. A total of 342 patients (105 dexmedetomidine and 237 propofol) were included in the analysis, with 190 matched (95 in each group) by propensity score. The primary outcome of this study was a composite of severe hypotension (mean arterial pressure < 60 mm Hg) and bradycardia (heart rate < 50 beats/min) during sedative infusion. No difference in the primary composite outcome in both the unmatched (30% vs 30%, p = 0.94) or matched cohorts (28% vs 34%, p = 0.35) could be found. When analyzed separately, no differences could be found in the prevalence of severe hypotension or bradycardia in either the unmatched or matched cohorts. Severe hypotension and bradycardia occur at similar prevalence in neurocritical care patients who receive dexmedetomidine or propofol. Providers should similarly consider the likelihood of hypotension or bradycardia before starting either sedative.


Detection of telomerase in hepatocellular carcinomas using a PCR ELISA assay: comparison with hTR expression.
While telomerase is undetectable in most normal somatic tissues, telomerase activation has been detected in many immortal cell lines and various cancers. To investigate telomerase expression in hepatocellular carcinoma, and to assess the expression of the RNA component of telomerase, hTR. 39 hepatocellular carcinomas were studied using a telomerase polymerase chain reaction (PCR) enzyme linked immunosorbent assay, which does not require radioactive PCR amplification and yields a semiquantitative measurement. Expression of hTR was also assessed by a non-radioactive in situ hybridisation procedure. The correlations between these two markers and the clinicopathological data were analysed. Telomerase activity was detected in 23 of 39 hepatocellular carcinoma specimens (59%). Comparison of hepatocellular carcinoma with and without telomerase expression, or with high and low telomerase (10 cases v 13 cases), showed no differences in the principal clinicopathological data. Although median survival was lower in the group with detectable telomerase activity than in that with undetectable activity (510 v 720 days) the difference was not significant (log-rank test, p = 0.08). hTR expression was detected in 11 of 14 cases of hepatocellular carcinoma tested (78%) and in four of 12 samples of adjacent non-cancerous tissue (33%). Five tumours and four non-cancerous tissues were positive for hTR, whereas no telomerase activity was detected in these. The presence of telomerase activity in hepatocellular carcinomas is confirmed. No correlation was observed between clinicopathological data and telomerase expression in hepatocellular carcinoma, but survival seemed better in the absence of telomerase expression. hTR seems to be more widely expressed than telomerase.


Clinical indicators of treatment-resistant psychosis.
Around 30% of individuals with schizophrenia remain symptomatic and significantly impaired despite antipsychotic treatment and are considered to be treatment resistant. Clinicians are currently unable to predict which patients are at higher risk of treatment resistance. To determine whether genetic liability for schizophrenia and/or clinical characteristics measurable at illness onset can prospectively indicate a higher risk of treatment-resistant psychosis (TRP). In 1070 individuals with schizophrenia or related psychotic disorders, schizophrenia polygenic risk scores (PRS) and large copy number variations (CNVs) were assessed for enrichment in TRP. Regression and machine-learning approaches were used to investigate the association of phenotypes related to demographics, family history, premorbid factors and illness onset with TRP. Younger age at onset (odds ratio 0.94, P = 7.79 × 10-13) and poor premorbid social adjustment (odds ratio 1.64, P = 2.41 × 10-4) increased risk of TRP in univariate regression analyses. These factors remained associated in multivariate regression analyses, which also found lower premorbid IQ (odds ratio 0.98, P = 7.76 × 10-3), younger father's age at birth (odds ratio 0.97, P = 0.015) and cannabis use (odds ratio 1.60, P = 0.025) increased the risk of TRP. Machine-learning approaches found age at onset to be the most important predictor and also identified premorbid IQ and poor social adjustment as predictors of TRP, mirroring findings from regression analyses. Genetic liability for schizophrenia was not associated with TRP. People with an earlier age at onset of psychosis and poor premorbid functioning are more likely to be treatment resistant. The genetic architecture of susceptibility to schizophrenia may be distinct from that of treatment outcomes.


Role of the plasma contact system in the pathogenesis of experimental anti-GBM glomerulonephritis.
To study the participation of the Hageman factor-related contact system of plasma in the pathogenesis of glomerulonephritis (GN), an anti-BM GN was induced in a group of 10 normal Brown Norway rats and another of seven Brown Norway BN/Mai Pfd f rats. The latter strain is characterized by a congenital deficiency of plasma prekallikrein and of high-molecular weight kininogen, with lengthening of the activated partial thromboplastin time. In the deficient group, one animal developed crescents in less than 25% of glomeruli, five in 25-50% and one in 50-75%. In the group of normal rats, extracapillary proliferation was of greater severity: one animal showed crescents in less than 25% of glomeruli, two in 50-75% and five in more than 75% of glomeruli. Although in both groups intense glomerular fibrin deposition was documented, the intensity of these deposits was less severe in the deficient animals. These data suggest, in the first place, that functional integrity of the contact system is not a necessary requirement for glomerular fibrinogenesis, other mechanisms being implicated in this phenomenon. On the other hand, this functional deficit has exerted a protective effect on crescent formation, which suggests that the contact system can play a role as a mediator of injury in glomerulonephritis, perhaps through the release of contact system-dependent mediators of inflammation.


A Cross-Sectional Study of Women and Girls with Congenital Bleeding Disorders: The American Thrombosis and Hemostasis Network Cohort.
Introduction: The number of women and girls (WG) with bleeding disorders cared for at hemophilia treatment centers has increased dramatically over the last 30 years, owing to improved recognition of bleeding symptoms specific to WG. However, basic epidemiologic data of this population remain elusive. The ATHNdataset (American Thrombosis and Hemostasis Network) is a surveillance tool for people with bleeding disorders in the United States, providing demographic as well as bleeding symptom and treatment information. The aim of this study was to characterize the female cohort within the ATHNdataset. Methods: In this retrospective cohort study, the ATHNdataset was queried for demographic data, bleeding disorder diagnosis, bleeding symptoms, and treatment. Descriptive statistics were used. Results: As of December 31, 2017, 8,820 WG with a congenital bleeding disorder were enrolled in the ATHNdataset, comprising 24.5% of the entire ATHNdataset cohort (35,945). The most common reported diagnosis was von Willebrand disease (VWD), accounting for 62.9% of the population. Reproductive tract bleeding was reported in 15.8% of participants older than 15 years. Conclusions: The ATHNdataset describes the largest cohort of WG with bleeding disorders to date. VWD is the most common diagnosis in WG with bleeding disorders. Symptoms specific to WG, such as heavy menstrual bleeding, are underreported in this data set compared with other data sources. Ongoing efforts are needed to improve diagnosis, treatment, and surveillance of WG with bleeding disorders.


Immunogenicity and safety of different dose schedules and antigen doses of an MF59-adjuvanted H7N9 vaccine in healthy adults aged 65 years and older.
The number of human influenza A (H7N9) infections has escalated since 2013 with high resultant mortality. We conducted a phase II, randomized, partially-blinded trial to evaluate the safety and immunogenicity of an MF59-adjuvanted inactivated, split virion, H7N9 influenza vaccine (H7N9 IIV) administered at various dose levels and schedules in older adults. 479 adults ≥ 65 years of age in stable health were randomized to one of six groups to receive either 3.75, 7.5 or 15 µg of influenza A/Shanghai/02/2013 (H7N9) IIV adjuvanted with MF59 given as a 3-dose series either on days 1, 28 and 168 or on days 1, 57 and 168. Immunogenicity was assessed using both hemagglutination inhibition (HAI) and microneutralization (MN) assays prior to and 28 days following each dose. Safety was assessed through 1 year following the last dose. Subjects in all groups had only modest immune responses, with the HAI GMT < 20 after the second vaccine dose and <29 after the third vaccine dose. HAI titers ≥ 40 were seen in <37% of subjects after the second dose and <49% after the third dose. There were no significant differences seen between the two dose schedules. MN titers followed similar patterns, although the titers were approximately two-fold higher than the HAI titers. Logistic regression modeling demonstrated no statistically significant associations between the immune responses and age, sex or body mass index whereas recent prior receipt of seasonal influenza vaccine significantly reduced the HAI response [OR 0.13 (95% CI 0.05, 0.33); p < 0.001]. Overall, the vaccine was well tolerated. Two mild potentially immune mediated adverse events occurred, lichen planus and guttate psoriasis. MF59-adjuvanted H7N9 IIV was only modestly immunogenic in the older adult population following three doses. There were no significant differences in antibody responses noted among the various antigen doses or the two dose schedules.


Genetic heterogeneity of induced pluripotent stem cells: results from 24 clones derived from a single C57BL/6 mouse.
Induced pluripotent stem cells (iPSCs) have tremendous potential as a tool for disease modeling, drug testing, and other applications. Since the generation of iPSCs "captures" the genetic history of the individual cell that was reprogrammed, iPSC clones (even those derived from the same individual) would be expected to demonstrate genetic heterogeneity. To assess the degree of genetic heterogeneity, and to determine whether some cells are more genetically "fit" for reprogramming, we performed exome sequencing on 24 mouse iPSC clones derived from skin fibroblasts obtained from two different sites of the same 8-week-old C57BL/6J male mouse. While no differences in the coding regions were detected in the two parental fibroblast pools, each clone had a unique genetic signature with a wide range of heterogeneity observed among the individual clones: a total of 383 iPSC variants were validated for the 24 clones (mean 16.0/clone, range 0-45). Since these variants were all present in the vast majority of the cells in each clone (variant allele frequencies of 40-60% for heterozygous variants), they most likely preexisted in the individual cells that were reprogrammed, rather than being acquired during reprogramming or cell passaging. We then tested whether this genetic heterogeneity had functional consequences for hematopoietic development by generating hematopoietic progenitors in vitro and enumerating colony forming units (CFUs). While there was a range of hematopoietic potentials among the 24 clones, only one clone failed to differentiate into hematopoietic cells; however, it was able to form a teratoma, proving its pluripotent nature. Further, no specific association was found between the mutational spectrum and the hematopoietic potential of each iPSC clone. These data clearly highlight the genetic heterogeneity present within individual fibroblasts that is captured by iPSC generation, and suggest that most of the changes are random, and functionally benign.


Reduced expression of manganese superoxide dismutase mRNA may correlate with invasiveness in esophageal carcinoma.
Little is known about the expression and antioxidant function of manganese superoxide dismutase (Mn-SOD) in esophageal squamous cell carcinoma. To determine the significance of Mn-SOD in esophageal squamous cell carcinomas, Mn-SOD mRNA expression was examined in 45 esophageal squamous cell carcinomas and the corresponding normal mucosal tissues by reverse transcription-polymerase chain reaction. The tumor/normal (T/N) ratio of 45 patients with esophageal carcinoma was calculated, and the data were clinicopathologically analyzed. The T/N ratio of Mn-SOD mRNA expression was less than 0.5 in 11 (32.4%) of 34 esophageal carcinoma cases without any preoperative treatments, while none of 11 cases who underwent preoperative chemotherapy showed a T/N ratio of <0.5 (p < 0.05). There was an inverse correlation between the Mn-SOD expression level and the degree of venous invasion (p < 0.05) as well as lymphatic invasion (p < 0.05). Furthermore, poorly differentiated squamous cell carcinoma showed significantly lower Mn-SOD mRNA expression levels than well differentiated carcinoma (p < 0.05). Our results suggest that Mn-SOD mRNA was frequently reduced in esophageal carcinoma when compared to the normal mucosa and the reduced expression levels of Mn-SOD mRNA may lead to an accumulation of superoxide radicals in conjunction with the increased invasiveness of esophageal carcinoma.


Stoichiometric modelling of assimilatory and dissimilatory biomass utilisation in a microbial community.
Assimilatory and dissimilatory utilisation of autotroph biomass by heterotrophs is a fundamental mechanism for the transfer of nutrients and energy across trophic levels. Metagenome data from a tractable, thermoacidophilic microbial community in Yellowstone National Park was used to build an in silico model to study heterotrophic utilisation of autotroph biomass using elementary flux mode analysis and flux balance analysis. Assimilatory and dissimilatory biomass utilisation was investigated using 29 forms of biomass-derived dissolved organic carbon (DOC) including individual monomer pools, individual macromolecular pools and aggregate biomass. The simulations identified ecologically competitive strategies for utilizing DOC under conditions of varying electron donor, electron acceptor or enzyme limitation. The simulated growth environment affected which form of DOC was the most competitive use of nutrients; for instance, oxygen limitation favoured utilisation of less reduced and fermentable DOC while carbon-limited environments favoured more reduced DOC. Additionally, metabolism was studied considering two encompassing metabolic strategies: simultaneous versus sequential use of DOC. Results of this study bound the transfer of nutrients and energy through microbial food webs, providing a quantitative foundation relevant to most microbial ecosystems.


Medical interns' reflections on their training in use of personal protective equipment.
The current COVID-19 pandemic has demonstrated that personal protective equipment (PPE) is essential, to prevent the acquisition and transmission of infectious diseases, yet its use is often sub-optimal in the clinical setting. Training and education are important to ensure and sustain the safe and effective use of PPE by medical interns, but current methods are often inadequate in providing the relevant knowledge and skills. The purpose of this study was to explore medical graduates' experiences of the use of PPE and identify opportunities for improvement in education and training programmes, to improve occupational and patient safety. This study was undertaken in 2018 in a large tertiary-care teaching hospital in Sydney, Australia, to explore medical interns' self-reported experiences of PPE use, at the beginning of their internship. Reflexive groups were conducted immediately after theoretical and practical PPE training, during hospital orientation. Transcripts of recorded discussions were analysed, using a thematic approach that drew on the COM-B (capability, opportunity, motivation - behaviour) framework for behaviour. 80% of 90 eligible graduates participated. Many interns had not previously received formal training in the specific skills required for optimal PPE use and had developed potentially unsafe habits. Their experiences as medical students in clinical areas contrasted sharply with recommended practice taught at hospital orientation and impacted on their ability to cultivate correct PPE use. Undergraduate teaching should be consistent with best practice PPE use, and include practical training that embeds correct and safe practices.


Smad signaling dynamics: insights from a parsimonious model.
The molecular mechanisms that transmit information from cell surface receptors to the nucleus are exceedingly complex; thus, much effort has been expended in developing computational models to understand these processes. A recent study on modeling the nuclear-cytoplasmic shuttling of Smad2-Smad4 complexes in response to transforming growth factor-beta (TGF-beta) receptor activation has provided substantial insight into how this signaling network translates the degree of TGF-beta receptor activation (input) into the amount of nuclear Smad2-Smad4 complexes (output). The study addressed this question by combining a simple, mechanistic model with targeted experiments, an approach that proved particularly powerful for exploring the fundamental properties of a complex signaling network. The mathematical model revealed that Smad nuclear-cytoplasmic dynamics enables a proportional but time-delayed coupling between the input and the output. As a result, the output can faithfully track gradual changes in the input while the rapid input fluctuations that constitute signaling noise are dampened out.


Clinical effectiveness and cost-effectiveness of the Rehabilitation Enablement in Chronic Heart Failure (REACH-HF) facilitated self-care rehabilitation intervention in heart failure patients and caregivers: rationale and protocol for a multicentre randomised controlled trial.
The Rehabilitation EnAblement in CHronic Heart Failure (REACH-HF) trial is part of a research programme designed to develop and evaluate a health professional facilitated, home-based, self-help rehabilitation intervention to improve self-care and health-related quality of life in people with heart failure and their caregivers. The trial will assess the clinical effectiveness and cost-effectiveness of the REACH-HF intervention in patients with systolic heart failure and impact on the outcomes of their caregivers. A parallel two group randomised controlled trial with 1:1 individual allocation to the REACH-HF intervention plus usual care (intervention group) or usual care alone (control group) in 216 patients with systolic heart failure (ejection fraction <45%) and their caregivers. The intervention comprises a self-help manual delivered by specially trained facilitators over a 12-week period. The primary outcome measure is patients' disease-specific health-related quality of life measured using the Minnesota Living with Heart Failure questionnaire at 12 months' follow-up. Secondary outcomes include survival and heart failure related hospitalisation, blood biomarkers, psychological well-being, exercise capacity, physical activity, other measures of quality of life, patient safety and the quality of life, psychological well-being and perceived burden of caregivers at 4, 6 and 12 months' follow-up. A process evaluation will assess fidelity of intervention delivery and explore potential mediators and moderators of changes in health-related quality of life in intervention and control group patients. Qualitative studies will describe patient and caregiver experiences of the intervention. An economic evaluation will estimate the cost-effectiveness of the REACH-HF intervention plus usual care versus usual care alone in patients with systolic heart failure. The study is approved by the North West-Lancaster Research Ethics Committee (ref 14/NW/1351). Findings will be disseminated via journals and presentations to publicise the research to clinicians, commissioners and service users. ISRCTN86234930; Pre-results.


Empirical Probability of Positive Response to PEEP Changes and Mechanical Ventilation Factors Associated With Improved Oxygenation During Pediatric Ventilation.
PEEP is titrated to improve oxygenation during mechanical ventilation. It is clinically desirable to identify factors that are associated with a clinical improvement or deterioration following a PEEP change. However, these factors have not been adequately described in the literature. Therefore, we aimed to quantify the empirical probability of PEEP changes having a positive effect upon oxygenation, compliance of the respiratory system (CRS), and the ratio of dead space to tidal volume (VD/VT). Further, clinical factors associated with positive response during pediatric mechanical ventilation are described. Mechanically ventilated pediatric subjects in the ICU were eligible for inclusion in the study. During PEEP increases (PEEPincrease), a responder was defined as having an improved SpO2 /FIO2 ratio; non-responders demonstrated a worsening SpO2 /FIO2 ratio in the following hour. When PEEP was decreased (PEEPdecrease), a responder was anyone who maintained or increased the SpO2 /FIO2 ratio; non-responders demonstrated a worsening SpO2 /FIO2 ratio. Features from continuous mechanical ventilation variables were extracted, and differences between responders and non-responders were identified. 286 PEEP change cases were eligible for analysis in 76 subjects. For PEEPincrease cases, the empirical probability of positive response was 56%, 67%, and 54% for oxygenation, CRS, and VD/VT, respectively. The median SpO2 /FIO2 increase was 13. For PEEPdecrease, the empirical probability of response was 46%, 53%, and 46% for oxygenation, CRS, and VD/VT, respectively. PEEPincrease responders had higher FIO2 requirements (70.8 vs 52.5%, P < .001), mean airway pressure (14.0 vs 12.9 cm H2O, P = .03), and oxygen saturation index (9.9 vs 7.5, P = .002) versus non-responders. For PEEPdecrease, VD/VT was lower in responders (0.46 vs 0.50, P = .031). In children requiring mechanical ventilation, the responder rate was modest for both PEEPincrease and PEEPdecrease cases. These data suggest that PEEP titration often does not have the desired clinical effect, and predicting which patients will manifest a positive response is complex, requiring more sophisticated means of assessing individual subjects.


Screening for colorectal cancer. A comparison of 3 fecal occult blood tests.
Colorectal cancer is the second leading cause of cancer deaths in the United States. Fecal occult blood testing has become a standard screening test for large-bowel cancers in the average asymptomatic population. Performance characteristics of the test and physician and participant compliance are the 2 major elements that impact the success of screening and early detection. To evaluate the nonhydrated Hemoccult, rehydrated Hemoccult, and Hemoccult SENSA tests (SmithKline Diagnostics Inc, Palo Alto, Calif) and to assess participant and physician compliance. A mass community-based screening study in an urban setting. Kits were distributed by a local pharmacy and at community sites. Diagnostic tests were completed through physicians' offices and clinics. Participants were asymptomatic and aged 50 years or older. Those who tested positive were advised to follow up with a physician. An overall positivity rate of 16% was reported for the 8293 kits that were processed. Rehydrated Hemoccult had a positivity rate of 15%; Hemoccult SENSA, 7%; and nonhydrated Hemoccult, 5%. The positive predictive value of nonhydrated Hemoccult was 14%; rehydrated Hemoccult, 7%; and Hemoccult SENSA, 11%. Of those who tested positive, 59% had a colonoscopy or flexible sigmoidoscopy and double-contrast barium enema examination on follow-up. Recommended follow-up was more frequent for those who consulted a gastroenterologist. Rehydrated Hemoccult yielded a higher positivity rate and lower positive predictive value than either Hemoccult SENSA or nonhydrated Hemoccult. Hemoccult SENSA approached the positive predictive value of nonhydrated Hemoccult. Adequacy of follow-up of patients testing positive for fecal occult blood needs improvement.


Integrating reinforcement learning, equilibrium points, and minimum variance to understand the development of reaching: a computational model.
Despite the huge literature on reaching behavior, a clear idea about the motor control processes underlying its development in infants is still lacking. This article contributes to overcoming this gap by proposing a computational model based on three key hypotheses: (a) trial-and-error learning processes drive the progressive development of reaching; (b) the control of the movements based on equilibrium points allows the model to quickly find the initial approximate solution to the problem of gaining contact with the target objects; (c) the request of precision of the end movement in the presence of muscular noise drives the progressive refinement of the reaching behavior. The tests of the model, based on a two degrees of freedom simulated dynamical arm, show that it is capable of reproducing a large number of empirical findings, most deriving from longitudinal studies with children: the developmental trajectory of several dynamical and kinematic variables of reaching movements, the time evolution of submovements composing reaching, the progressive development of a bell-shaped speed profile, and the evolution of the management of redundant degrees of freedom. The model also produces testable predictions on several of these phenomena. Most of these empirical data have never been investigated by previous computational models and, more important, have never been accounted for by a unique model. In this respect, the analysis of the model functioning reveals that all these results are ultimately explained, sometimes in unexpected ways, by the same developmental trajectory emerging from the interplay of the three mentioned hypotheses: The model first quickly learns to perform coarse movements that assure a contact of the hand with the target (an achievement with great adaptive value) and then slowly refines the detailed control of the dynamical aspects of movement to increase accuracy.


Esophageal cancer metabolite biomarkers detected by LC-MS and NMR methods.
Esophageal adenocarcinoma (EAC) is a rarely curable disease and is rapidly rising worldwide in incidence. Barret's esophagus (BE) and high-grade dysplasia (HGD) are considered major risk factors for invasive adenocarcinoma. In the current study, unbiased global metabolic profiling methods were applied to serum samples from patients with EAC, BE and HGD, and healthy individuals, in order to identify metabolite based biomarkers associated with the early stages of EAC with the goal of improving prognostication. Serum metabolite profiles from patients with EAC (n = 67), BE (n = 3), HGD (n = 9) and healthy volunteers (n = 34) were obtained using high performance liquid chromatography-mass spectrometry (LC-MS) methods. Twelve metabolites differed significantly (p<0.05) between EAC patients and healthy controls. A partial least-squares discriminant analysis (PLS-DA) model had good accuracy with the area under the receiver operative characteristic curve (AUROC) of 0.82. However, when the results of LC-MS were combined with 8 metabolites detected by nuclear magnetic resonance (NMR) in a previous study, the combination of NMR and MS detected metabolites provided a much superior performance, with AUROC = 0.95. Further, mean values of 12 of these metabolites varied consistently from healthy controls to the high-risk individuals (BE and HGD patients) and EAC subjects. Altered metabolic pathways including a number of amino acid pathways and energy metabolism were identified based on altered levels of numerous metabolites. Metabolic profiles derived from the combination of LC-MS and NMR methods readily distinguish EAC patients and potentially promise important routes to understanding the carcinogenesis and detecting the cancer. Differences in the metabolic profiles between high-risk individuals and the EAC indicate the possibility of identifying the patients at risk much earlier to the development of the cancer.


Roles for endothelial cell and macrophage Gch1 and tetrahydrobiopterin in atherosclerosis progression.
GTP cyclohydrolase I catalyses the first and rate-limiting reaction in the synthesis of tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthases (NOS). Both eNOS and iNOS have been implicated in the progression of atherosclerosis, with opposing effects in eNOS and iNOS knockout mice. However, the pathophysiologic requirement for BH4 in regulating both eNOS and iNOS function, and the effects of loss of BH4 on the progression of atherosclerosis remains unknown. Hyperlipidemic mice deficient in Gch1 in endothelial cells and leucocytes were generated by crossing Gch1fl/flTie2cre mice with ApoE-/- mice. Deficiency of Gch1 and BH4 in endothelial cells and myeloid cells was associated with mildly increased blood pressure. High fat feeding for 6 weeks in Gch1fl/flTie2CreApoE-/- mice resulted in significantly decreased circulating BH4 levels, increased atherosclerosis burden and increased plaque macrophage content. Gch1fl/flTie2CreApoE-/- mice showed hallmarks of endothelial cell dysfunction, with increased aortic VCAM-1 expression and decreased endothelial cell dependent vasodilation. Furthermore, loss of BH4 from pro-inflammatory macrophages resulted in increased foam cell formation and altered cellular redox signalling, with decreased expression of antioxidant genes and increased reactive oxygen species. Bone marrow chimeras revealed that loss of Gch1 in both endothelial cells and leucocytes is required to accelerate atherosclerosis. Both endothelial cell and macrophage BH4 play important roles in the regulation of NOS function and cellular redox signalling in atherosclerosis.


alpha- and beta-adrenergic control of thermogenin mRNA expression in brown adipose tissue.
By the use of an earlier characterised cDNA clone, CIN-1, corresponding to a sequence of the mRNA coding for the brown-fat specific "uncoupling" protein, thermogenin, the amount of thermogenin mRNA found in the brown adipose tissue of mice was quantitatively investigated under different physiological and pharmacological conditions. It was found that a 4 hr cold stress led to a 7-fold increase in the amount of thermogenin mRNA; injection of norepinephrine had a significant but smaller effect. Most notably, isoprenaline (beta-agonist) and phenylephrine (alpha-agonist) had in themselves no effect, but when injected together were able to increase the mRNA level synergistically. In 4 hr cold-stressed mice, norepinephrine, isoprenaline and cholera toxin could all further potentiate the effect of the cold stress itself on the mRNA level. Insulin and the glucocorticoid dexamethasone both had weak stimulatory effects on the mRNA level. It is concluded that an increase in intracellular cAMP levels is a necessary and perhaps sufficient stimulus for the increase in thermogenin gene expression. However, at least under in vivo conditions, this increase requires stimulation of both alpha- and beta-adrenergic pathways.


Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestris pv. vesicatoria.
Specific cDNAs showing differential expression in bacteria-infected pepper leaves as opposed to healthy leaves were isolated from a pepper cDNA library from hypersensitive response (HR) lesions of leaves infected with an avirulent strain of Xanthomonas campestris pv. vesicatoria. Among a total of 282 cDNA clones tested, 36 individual cDNA genes (13%) hybridized strongly or differentially to the cDNA probes from bacteria-infected leaves. Ten Capsicum Annuum-Induced (CAI) genes encoding putative thionin, lipid transfer protein I and II, osmotin (PR-5), class I chitinase, beta-1,3-glucanase, SAR 8.2, stellacyanin, leucine-rich repeat protein, and auxin-repressed protein were identified. Two CAI genes showed little or no sequence homology to the previously sequenced plant genes. Transcripts of the CAI genes were strongly or preferentially induced in pepper tissues by infection with X. campestris pv. vesicatoria or Phytophthora capsici, and by abiotic elicitor treatment. In particular, most of the CAI genes were strongly induced in pepper tissues by ethephon and methyl jasmonate.


Conference summary. Directions and needs in asbestos research: new insights.
The Center for Environmental Health Sciences (CEHS) Conference entitled "Directions and Needs in Asbestos Research: New Insights" was held at the University of Montana in Missoula on July 28 and 29, 2005. Researchers, physicians, health care workers, and federal agency representatives from around the country met for a cross-disciplinary exploration of many issues related to asbestos research. Topics included community and psychosocial issues in biomedical research, asbestos exposure assessment, assessment and mechanisms of asbestos-related diseases, and new research directions. This meeting report is a summary of the conference presentations, and of the topics identified for future research directions. This conference was a follow-up to one hosted by the CEHS in June 2002, and continued to take advantage of opportunities to work with a unique population in Libby, MT, where significant asbestos exposures have occurred due to the mining of asbestos-contaminated vermiculite. The goals of this conference were to bring together experts from diverse fields to identify progress made since the last conference and to develop new research avenues that would allow us to address the research needs in emerging asbestos-exposed populations. Participants indicated that these objectives were met, and expressed enthusiasm for follow-up conferences, tentatively planned for the summer of 2007, to maintain the dialog that has been established regarding directions and needs in asbestos research. Selected papers from the conference are presented in this issue of Inhalation Toxicology.


Risk of intestinal helminth and protozoan infection in a refugee population.
With continuing emigration from endemic countries, screening for parasitic infections remains a priority in U.S. communities serving refugee and immigrant populations. We report the prevalence of helminths and protozoa as well as demographic risk factors associated with these infections among 533 refugees seen at the Santa Clara County, California, Refugee Clinic between October 2001 and January 2004. Stool parasites were identified from 14% of refugees, including 9% found to have one or more protozoa and 6% found to have at least one helminth. Most common protozoan infections were Giardia lamblia (6%) and Dientamoeba fragilis (3%), and for helminths, hookworm (2%). Protozoa were more frequent in refugees < 18 years of age (OR: 2.2 [1.2-4.2]), whereas helminths were more common in refugees from South Central Asia (OR: 8.0 [2.3-27.7]) and Africa (OR: 5.9 [1.6-21.6]) when compared with refugees from Eastern Europe and the Middle East. Among helminths, Ascaris lumbricoides and hookworm were concentrated among South Central Asians (6 of 7 and 10 of 11 cases, respectively), whereas Strongyloides stercoralis was predominantly found in Africans (5 of 7 cases). Although predeparture empirical treatment programs in Saharan Africa may have helped to reduce prevalence among arriving refugees from this region, parasitic infection is still common among refugees to the United States with helminth infections found in more specific populations. As refugees represent only a fraction of recent immigrants from endemic countries, current studies in nonrefugee groups are also needed.


Charcot Midfoot Reconstruction: Does Subtalar Arthrodesis or Medial Column Fixation Improve Outcomes?
Charcot neuroarthropathy is a complication of neuropathy often secondary to diabetes mellitus and most commonly affects the midfoot. In these patients, reconstruction of the foot may be required for limb salvage. A superconstruct technique has previously been described using intramedullary beaming fixation of the midfoot and hindfoot to span the zone of injury. Inclusion of the subtalar joint in the arthrodesis construct is not consistently performed among different surgeons. The aim of this study was to describe midfoot beaming constructs and postoperative complications after midfoot reconstruction with and without subtalar arthrodesis. We reviewed medical records of patients who underwent midfoot Charcot reconstruction with an intramedullary beaming superconstruct. Patients included in the study had at least 3 months of follow-up and had Sanders-Frykberg II/III classification of Charcot neuroarthropathy. Postoperative radiographs were evaluated for evidence of hardware failure at the latest follow-up evaluation. The main variables of interest were: hardware failure or nonunion requiring revision operation, deep infection, and unplanned reoperation. Thirty patients who underwent midfoot reconstruction were included. The mean follow-up was 67.4 ± 25.9 weeks. Twenty-two (73.3%) patients had concomitant subtalar arthrodesis and midfoot beaming. Overall complications were lower in patients with subtalar arthrodesis (40.9%) than those without subtalar arthrodesis (75%) resulting in an odds ratio of 0.271 (0.042-1.338, p = .146). Furthermore, increased number of screws used in the midfoot construct was negatively correlated with complications (r = -0.44, p = .01). An intramedullary midfoot beaming superconstruct with subtalar arthrodesis has previously been proposed to provide better fixation after midfoot beaming Charcot neuroarthropathy reconstruction. Our results suggest including the subtalar joint as part of a superconstruct for the reconstruction of Sanders-Frykberg II/III Charcot results in an 80% lower complication rate than intramedullary beaming alone. We also found an increased number of screws used in the midfoot results in a lower complication rate.


PD-1 and PD-L1 Expression in NSCLC Indicate a Favorable Prognosis in Defined Subgroups.
Immunotherapy can become a crucial therapeutic option to improve prognosis for lung cancer patients. First clinical trials with therapies targeting the programmed cell death receptor PD-1 and its ligand PD-L1 have shown promising results in several solid tumors. However, in lung cancer the diagnostic, prognostic and predictive value of these immunologic factors remains unclear. The impact of both factors was evaluated in a study collective of 321 clinically well-annotated patients with non-small lung cancer (NSCLC) using immunohistochemistry. PD-1 expression by tumor infiltrating lymphocytes (TILs) was found in 22%, whereas tumor cell associated PD-L1 expression was observed in 24% of the NSCLC tumors. In Fisher's exact test a positive correlation was found for PD-L1 and Bcl-xl protein expression (p = 0.013). Interestingly, PD-L1 expression on tumor cells was associated with improved overall survival in pulmonary squamous cell carcinomas (SCC, p = 0.042, log rank test), with adjuvant therapy (p = 0.017), with increased tumor size (pT2-4, p = 0.039) and with positive lymph node status (pN1-3, p = 0.010). These observations were confirmed by multivariate cox regression models. One major finding of our study is the identification of a prognostic implication of PD-L1 in subsets of NSCLC patients with pulmonary SCC, with increased tumor size, with a positive lymph node status and NSCLC patients who received adjuvant therapies. This study provides first data for immune-context related risk stratification of NSCLC patients. Further studies are necessary both to confirm this observation and to evaluate the predictive value of PD-1 and PD-L1 in NSCLC in the context of PD-1 inhibition.


High-temperature superconductor bulk magnets that can trap magnetic fields of over 17 tesla at 29 K.
Large-grain high-temperature superconductors of the form RE-Ba-Cu-O (where RE is a rare-earth element) can trap magnetic fields of several tesla at low temperatures, and so can be used for permanent magnet applications. The magnitude of the trapped field is proportional to the critical current density and the volume of the superconductor. Various potential engineering applications for such magnets have emerged, and some have already been commercialized. However, the range of applications is limited by poor mechanical stability and low thermal conductivity of the bulk superconductors; RE-Ba-Cu-O magnets have been found to fracture during high-field activation, owing to magnetic pressure. Here we present a post-fabrication treatment that improves the mechanical properties as well as thermal conductivity of a bulk Y-Ba-Cu-O magnet, thereby increasing its field-trapping capacity. First, resin impregnation and wrapping the materials in carbon fibre improves the mechanical properties. Second, a small hole drilled into the centre of the magnet allows impregnation of Bi-Pb-Sn-Cd alloy into the superconductor and inclusion of an aluminium wire support, which results in a significant enhancement of thermal stability and internal mechanical strength. As a result, 17.24 T could be trapped, without fracturing, in a bulk Y-Ba-Cu-O sample of 2.65 cm diameter at 29 K.


Orthopedic Injuries in Professional Surfers: A Retrospective Study at a Single Orthopedic Center.
Literature regarding surfing injuries is scarce and most studies report a high number of minor injuries. Recent literature suggests that musculoskeletal injuries are more common than previously reported. In the mid 2000s, competitive surfing has seen an increase in aerial maneuvers, as they have become more highly rewarded by the judging panel, which may be contributing to a change in injury patterns. Our goal was to evaluate orthopedic injuries in professional surfers. Orthopedic injuries in professional surfers are diverse and have evolved with skill and technology. Retrospective observational study. Level 4 Case Series. Medical records of professional surfers from a single orthopedic center between 1991 and 2016 were reviewed. Injuries sustained while surfing and chronic injuries related to surfing were included. Site of injury, diagnosis, and treatment were recorded along with demographic data. In total, 163 injuries in 86 athletes were recorded. The median injury age was 28.5 years (range 12-62 years) and 92.6% of patients were male. The most frequently injured body parts were the knee (28.2%), ankle (22.1%), and shoulder (19%). Most knee injuries were of the medial collateral ligament at 49%, with 75% of ankle injuries being sprains, and in shoulder, 48% had instability, 42% had a rotator cuff tear, and 35% had a superior anterior-posterior labral tear. Lower extremity injuries primarily affected the surfers' back leg (72.5%). In all, 34.6% of all injuries required surgical intervention and involved shoulder injuries most the time. The most common orthopedic injuries in professional surfers involve the knee, ankle, shoulder, hip, and back. Surfers' rear extremities were preferentially injured which is the extremity responsible for power and torque. Shoulder injuries increased the probability of an operative intervention. Last, overuse injuries (femoral-acetabular impingement, rotator cuffs) occurred in the older surf population compared with more acute injuries (ankle sprains/fractures, anterior cruciate ligament tears) which is also consistent with time to surgery.


Long-term results of sealed capsule irrigation using distilled water to prevent posterior capsule opacification: a prospective clinical randomised trial.
We investigated long-term safety and efficacy of sealed capsule irrigation (SCI) during cataract surgery to prevent posterior capsule opacification (PCO). One eye of each of 17 patients (mean age: 70.1+/-9.7 years) who presented with bilateral cataracts was randomly chosen for SCI treatment. After phacoemulsification, the capsular bag was vacuum sealed with the PerfectCapsule device (Milvella) followed by SCI using distilled water for two minutes. No vacuum loss occurred during irrigation. Each patient's fellow eye served as a control. One hydrophilic acrylic intraocular lens model was implanted in all eyes. Five patients had to be excluded due to deep anterior chamber, small pupil or unilateral surgery. Follow-up examinations took place one day and one, three, six, 12 and 24 months after surgery. We evaluated safety parameters, anterior capsule (AC) overlapping and PCO. Postoperatively, mean best corrected visual acuity, pachymetry, endothelial cell count, intraocular pressure, AC overlapping and PCO showed no statistically significant difference between SCI and the control group (p>0.05, Wilcoxon test). SCI is a safe procedure and enables the specific pharmacological targeting of lens epithelial cells inside the capsular bag. Using distilled water, however, it is not possible to reduce PCO development significantly. Thus, alternative substances should be evaluated.


PET imaging reveals distinctive roles for different regional adipose tissue depots in systemic glucose metabolism in nonobese humans.
Excess amounts of abdominal subcutaneous (SAT) and visceral (VAT) adipose tissue (AT) are associated with insulin resistance, even in normal-weight subjects. In contrast, gluteal-femoral AT (GFAT) is hypothesized to offer protection against insulin resistance. Dynamic PET imaging studies were undertaken to examine the contributions of both metabolic activity and size (volume) of these depots in systemic glucose metabolism. Nonobese, healthy volunteers (n = 15) underwent dynamic PET imaging uptake of [¹⁸F]FDG at a steady-state (20 mU·m⁻²·min⁻¹) insulin infusion. PET images of tissue [¹⁸F]FDG activity were coregistered with MRI to derive K values for insulin-stimulated rates of fractional glucose uptake within tissue. Adipose tissue volume was calculated from DEXA and MRI. VAT had significantly higher rates of fractional glucose uptake per volume than SAT (P < 0.05) or GFAT (P < 0.01). K(GFAT) correlated positively (r = 0.67, P < 0.01) with systemic insulin sensitivity [glucose disappearance rate (R(d))] and negatively with insulin-suppressed FFA (r = -0.71, P < 0.01). SAT (r = -0.70, P < 0.01) and VAT mass (r = -0.55, P < 0.05) correlated negatively with R(d), but GFAT mass did not. We conclude that rates of fractional glucose uptake within GFAT and VAT are significantly and positively associated with systemic insulin sensitivity in nonobese subjects. Furthermore, whereas SAT and VAT amounts are confirmed to relate to systemic insulin resistance, GFAT amount is not associated with insulin resistance. These dynamic PET imaging studies indicate that both quantity and quality of specific AT depots have distinct roles in systemic insulin resistance and may help explain the metabolically obese but normal-weight phenotype.


Mycobacterium lutetiense sp. nov., Mycobacterium montmartrense sp. nov. and Mycobacterium arcueilense sp. nov., members of a novel group of non-pigmented rapidly growing mycobacteria recovered from a water distribution system.
From our recent survey of non-pigmented rapidly growing mycobacteria in the Parisian water system, three groups of isolates (taxons 1-3) corresponding to possible novel species were selected for taxonomic study. The three taxa each formed creamy white, rough colonies, had an optimal growth temperature of 30 °C, hydrolyzed Tween 80, were catalase-positive at 22 °C and expressed arylsulfatase activity. All three were susceptible to amikacin, ciprofloxacin and tigecycline. The three taxa produced specific sets of mycolic acids, including one family that has never previously been described, as determined by thin layer chromatography and nuclear magnetic resonance. The partial rpoB sequences (723 bp) showed 4-6 % divergence from each other and more than 5 % differences from the most similar species. Partial 16S rRNA gene sequences showed 99 % identity within each species. The most similar sequences for 16S rRNA genes (98-99 % identity over 1444-1461 bp) were found in the Mycobacterium fortuitum group, Mycobacterium septicum and Mycobacterium farcinogenes. The three taxa formed a new clade (bootstrap value, 99 %) on trees reconstructed from concatenated partial 16S rRNA, hsp65 and rpoB sequences. The above results led us to propose three novel species for the three groups of isolates, namely Mycobacterium lutetiense sp. nov. [type strain 071T=ParisRGMnew_1T (CIP 110656T=DSM 46713T)], Mycobacterium montmartrense sp. nov. [type strain 196T=ParisRGMnew_2T (CIP 110655T=DSM 46714T)] and Mycobacteriu marcueilense sp. nov. [type strain of 269T=ParisRGMnew_3T (CIP 110654T=DSM 46715T)].


People's beliefs about factors contributing to mental health: implications for mental health promotion.
To quantify people's perceptions of mental health identified in qualitative research and to inform mental health promotion communication strategies. A statewide telephone survey of 1,500 adults was conducted in Western Australia using a structured questionnaire containing both open and closed-ended questions. The vast majority of people had negative (or illness) connotations to the words 'mental health', but had positive connotations to the term 'mentally healthy person'. The three factors perceived to contribute most to being mentally healthy were: having good friends to talk problems over with; keeping one's mind active; and the opportunity to have control over one's life. The three factors perceived to contribute most to being mentally unhealthy were: excessive use of alcohol or drugs; having no friends or support network; and life crises o traumas. The phrase 'being content with who you are' best summed up good mental health. Older people generally placed greater emphasis than younger people on cognitive functioning and keeping physically healthy for good mental health. People's beliefs about factors influencing mental health are consistent with much of the literature. Communication components of mental health promotion interventions based on the data reported here would be viewed as credible and relevant by most people.


OMIC Technologies and Vaccine Development: From the Identification of Vulnerable Individuals to the Formulation of Invulnerable Vaccines.
Routine vaccination is among the most effective clinical interventions to prevent diseases as it is estimated to save over 3 million lives every year. However, the full potential of global immunization programs is not realised because population coverage is still suboptimal. This is also due to the inadequate immune response and paucity of informative correlates of protection upon immunization of vulnerable individuals such as newborns, preterm infants, pregnant women, and elderly individuals as well as those patients affected by chronic and immune compromising medical conditions. In addition, these groups are undervaccinated for a number of reasons, including lack of awareness of vaccine-preventable diseases and uncertainty or misconceptions about the safety and efficacy of vaccination by parents and healthcare providers. The presence of these nonresponders/undervaccinated individuals represents a major health and economic burden to society, which will become particularly difficult to address in settings with limited public resources. This review describes innovative and experimental approaches that can help identify specific genomic profiles defining nonresponder individuals for whom specific interventions might be needed. We will provide examples that show how such information can be useful to identify novel biomarkers of safety and immunogenicity for future vaccine trials. Finally, we will discuss how system biology "OMICs" data can be used to design bioinformatic tools to predict the vaccination outcome providing genetic and molecular "signatures" of protective immune response. This strategy may soon enable identification of signatures highly predictive of vaccine safety, immunogenicity, and efficacy/protection thereby informing personalized vaccine interventions in vulnerable populations.


Independent effects of attentional gain control and competitive interactions on visual stimulus processing.
Attention filters behaviorally relevant stimuli from the constant stream of sensory information comprising our environment. Research into underlying neural mechanisms in humans suggests that visual attention biases mutual suppression between stimuli resulting from competition for limited processing resources. As a consequence, processing of an attended stimulus is facilitated. This account makes 2 assumptions: 1) An attended stimulus is released from mutual suppression with competing stimuli and 2) an attended stimulus experiences greater gain in the presence of competing stimuli than when it is presented alone. Here, we tested these assumptions by recording frequency-tagged potentials elicited in early visual cortex that index stimulus-specific processing. We contrasted the processing of a given stimulus when its location was attended or unattended and in the presence or the absence of a nearby competing stimulus. At variance with previous findings, competition similarly suppressed processing of attended and unattended stimuli. Moreover, the magnitude of attentional gain was comparable in the presence or the absence of competing stimuli. We conclude that visuospatial selective attention does not directly modulate mutual suppression between stimuli but instead acts as a signal gain, which biases processing toward attended stimuli independent of competition.


Situating stress: lessons from lay discourses on diabetes.
In response to the serious toll diabetes takes on health and resources, researchers increasingly are examining physical and psychological pathways that affect and are affected by diabetes, including stress. Although biomedical researchers and practitioners are beginning to recognize the association between stress and diabetes onset and management, laypersons have long-standing and extensive insights into the multiple ways in which stress is associated with the diabetes disease process. In this article, we examine lay perspectives on stress and diabetes among a multiethnic sample of 80 adults. Participants suggest varying arenas in which stress intersects with diabetes, including stress as implicated in the origin of diabetes, as a threat to maintaining glycemic control, as a challenge to self-management, and as a precursor to and a consequence of diabetes complications. An improved understanding of such perspectives may enhance appropriate disease management and develop a more valid conceptualization of stress in research efforts.


Lysine methylation promotes VEGFR-2 activation and angiogenesis.
Activation of vascular endothelial growth factor receptor-2 (VEGFR-2), an endothelial cell receptor tyrosine kinase, promotes tumor angiogenesis and ocular neovascularization. We report the methylation of VEGFR-2 at multiple Lys and Arg residues, including Lys(1041), a residue that is proximal to the activation loop of the kinase domain. Methylation of VEGFR-2 was independent of ligand binding and was not regulated by ligand stimulation. Methylation of Lys(1041) enhanced tyrosine phosphorylation and kinase activity in response to ligands. Additionally, interfering with the methylation of VEGFR-2 by pharmacological inhibition or by site-directed mutagenesis revealed that methylation of Lys(1041) was required for VEGFR-2-mediated angiogenesis in zebrafish and tumor growth in mice. We propose that methylation of Lys(1041) promotes the activation of VEGFR-2 and that similar posttranslational modification could also regulate the activity of other receptor tyrosine kinases.


Impact of metformin on reproductive tissues: an overview from gametogenesis to gestation.
Metformin is an oral anti-hyperglycemic drug that acts as an insulin sensitizer in the treatment of diabetes mellitus type 2. It has also been widely used in the treatment of polycystic ovary syndrome (PCOS) and gestational diabetes. This drug has been shown to activate a protein kinase called 5' AMP-activated protein kinase or AMPK. AMPK is present in many tissues making metformin's effect multi factorial. However as metformin crosses the placenta, its use during pregnancy raises concerns regarding potential adverse effects on the mother and fetus. The majority of reports suggest no significant adverse effects or teratogenicity. However, disconcerting reports of male mouse offspring that were exposed to metformin in utero that present with a reduction in testis size, seminiferous tubule size and in Sertoli cell number suggest that we do not understand the full suite of effects of metformin. In addition, recent molecular evidence is suggesting an epigenetic effect of metformin which could explain some of the long-term effects reported. Nevertheless, the data are still insufficient to completely confirm or disprove negative effects of metformin. The aims of this review are to provide a summary of the safety of metformin in various aspects of sexual reproduction, the use of metformin by gestating mothers, and its possible side-effects on offspring from women who are administered metformin during pregnancy.


Forest dynamics in the U.S. indicate disproportionate attrition in western forests, rural areas and public lands.
Forests are experiencing significant changes; studying geographic patterns in forests is critical in understanding the impact of forest dynamics to biodiversity, soil erosion, water chemistry and climate. Few studies have examined forest geographic pattern changes other than fragmentation; however, other spatial processes of forest dynamics are of equal importance. Here, we study forest attrition, the complete removal of forest patches, that can result in complete habitat loss, severe decline of population sizes and species richness, and shifts of local and regional environmental conditions. We aim to develop a simple yet insightful proximity-based spatial indicator capturing forest attrition that is independent of spatial scale and boundaries with worldwide application potential. Using this proximity indicator, we evaluate forest attrition across ecoregions, land ownership and urbanization stratifications across continental United States of America. Nationally, the total forest cover loss was approximately 90,400 km2, roughly the size of the state of Maine, constituting a decline of 2.96%. Examining the spatial arrangement of this change the average FAD was 3674m in 1992 and increased by 514m or 14.0% in 2001. Simulations of forest cover loss indicate only a 10m FAD increase suggesting that the observed FAD increase was more than an order of magnitude higher than expected. Furthermore, forest attrition is considerably higher in the western United States, in rural areas and in public lands. Our mathematical model (R2 = 0.93) supports estimation of attrition for a given forest cover. The FAD metric quantifies forest attrition across spatial scales and geographic boundaries and assesses unambiguously changes over time. The metric is applicable to any landscape and offers a new complementary insight on forest landscape patterns from local to global scales, improving future exploration of drivers and repercussions of forest cover changes and supporting more informative management of forest carbon, changing climate and species biodiversity.


Dendritic Cells Generated From Mops condylurus, a Likely Filovirus Reservoir Host, Are Susceptible to and Activated by Zaire Ebolavirus Infection.
Ebola virus infection of human dendritic cells (DCs) induces atypical adaptive immune responses and thereby exacerbates Ebola virus disease (EVD). Human DCs, infected with Ebola virus aberrantly express low levels of the DC activation markers CD80, CD86, and MHC class II. The T cell responses ensuing are commonly anergic rather than protective against EVD. We hypothesize that DCs derived from potential reservoir hosts such as bats, which do not develop disease signs in response to Ebola virus infection, would exhibit features associated with activation. In this study, we have examined Zaire ebolavirus (EBOV) infection of DCs derived from the Angolan free-tailed bat species, Mops condylurus. This species was previously identified as permissive to EBOV infection in vivo, in the absence of disease signs. M. condylurus has also been recently implicated as the reservoir host for Bombali ebolavirus, a virus species that is closely related to EBOV. Due to the absence of pre-existing M. condylurus species-specific reagents, we characterized its de novo assembled transcriptome and defined its phylogenetic similarity to other mammals, which enabled the identification of cross-reactive reagents for M. condylurus bone marrow-derived DC (bat-BMDC) differentiation and immune cell phenotyping. Our results reveal that bat-BMDCs are susceptible to EBOV infection as determined by detection of EBOV specific viral RNA (vRNA). vRNA increased significantly 72 h after EBOV-infection and was detected in both cells and in culture supernatants. Bat-BMDC infection was further confirmed by the observation of GFP expression in DC cultures infected with a recombinant GFP-EBOV. Bat-BMDCs upregulated CD80 and chemokine ligand 3 (CCL3) transcripts in response to EBOV infection, which positively correlated with the expression levels of EBOV vRNA. In contrast to the aberrant responses to EBOV infection that are typical for human-DC, our findings from bat-BMDCs provide evidence for an immunological basis of asymptomatic EBOV infection outcomes.


The effect of cyclic loading simulating oscillatory joint mobilization on the posterior capsule of the glenohumeral joint: a cadaveric study.
Experimental laboratory design. To examine the effect of force and repetition during oscillatory joint mobilizations on the posterior capsule of the glenohumeral joint. The optimal external force and frequency to be used during joint mobilization to elongate the posterior capsule of the glenohumeral joint has yet to be identified. Twenty-one posterior capsules were harvested from fresh-frozen shoulders. A cyclic loading test simulating oscillatory posterior joint mobilization on the shoulder specimens was performed with a material testing machine. The specimens were assigned to 3 different loading groups simulating joint mobilization in the toe (5 N), transition (20 N), and beginning of the linear regions (40 N) of the load displacement curve. Displacement of the humeral head at an applied load of 5 N was recorded at the 1st, 100th, 200th, 300th, 400th, 500th, and 600th cycles and at 1 hour after completion of the cyclic loading. Furthermore, stiffness was calculated after the 1st and 600th cycles and 1 hour after testing. Humeral head displacement was significantly greater for the 100th to 600th cycle, compared to the 1st cycle, for all 3 loading groups. Significant increases in displacement and stiffness were observed between the 1st cycle and 1 hour after completion of the cyclic tests for both the 20-N and 40-N loading groups. While oscillatory joint mobilization to a force of 5 N resulted in temporary elongation of the posterior capsule, mobilization to loads of 20 and 40 N resulted in sustained elongation of the capsule for up to 1 hour. Our findings also suggest that mobilization up to loads that represent the beginning of the linear region of the load displacement curve could be performed without serious damage to the posterior capsule.


ECHS1 mutations in Leigh disease: a new inborn error of metabolism affecting valine metabolism.
Two siblings with fatal Leigh disease had increased excretion of S-(2-carboxypropyl)cysteine and several other metabolites that are features of 3-hydroxyisobutyryl-CoA hydrolase (HIBCH) deficiency, a rare defect in the valine catabolic pathway associated with Leigh-like disease. However, this diagnosis was excluded by HIBCH sequencing and normal enzyme activity. In contrast to HIBCH deficiency, the excretion of 3-hydroxyisobutyryl-carnitine was normal in the children, suggesting deficiency of short-chain enoyl-CoA hydratase (ECHS1 gene). This mitochondrial enzyme is active in several metabolic pathways involving fatty acids and amino acids, including valine, and is immediately upstream of HIBCH in the valine pathway. Both children were compound heterozygous for a c.473C > A (p.A158D) missense mutation and a c.414+3G>C splicing mutation in ECHS1. ECHS1 activity was markedly decreased in cultured fibroblasts from both siblings, ECHS1 protein was undetectable by immunoblot analysis and transfection of patient cells with wild-type ECHS1 rescued ECHS1 activity. The highly reactive metabolites methacrylyl-CoA and acryloyl-CoA accumulate in deficiencies of both ECHS1 and HIBCH and are probably responsible for the brain pathology in both disorders. Deficiency of ECHS1 or HIBCH should be considered in children with Leigh disease. Urine metabolite testing can detect and distinguish between these two disorders.


Structural basis for substrate binding and catalytic mechanism of a human RNA:m5C methyltransferase NSun6.
5-methylcytosine (m5C) modifications of RNA are ubiquitous in nature and play important roles in many biological processes such as protein translational regulation, RNA processing and stress response. Aberrant expressions of RNA:m5C methyltransferases are closely associated with various human diseases including cancers. However, no structural information for RNA-bound RNA:m5C methyltransferase was available until now, hindering elucidation of the catalytic mechanism behind RNA:m5C methylation. Here, we have solved the structures of NSun6, a human tRNA:m5C methyltransferase, in the apo form and in complex with a full-length tRNA substrate. These structures show a non-canonical conformation of the bound tRNA, rendering the base moiety of the target cytosine accessible to the enzyme for methylation. Further biochemical assays reveal the critical, but distinct, roles of two conserved cysteine residues for the RNA:m5C methylation. Collectively, for the first time, we have solved the complex structure of a RNA:m5C methyltransferase and addressed the catalytic mechanism of the RNA:m5C methyltransferase family, which may allow for structure-based drug design toward RNA:m5C methyltransferase-related diseases.


Epidemiological characteristics of human stampedes.
The potential for deadly human stampedes to occur at any mass gathering event highlights this unique form of crowd disaster as deserving of special attention from both scientific and planning perspectives. Improved understanding of human stampedes is indispensable in the mitigation of this type of mass casualty. With relatively few peer-reviewed reports on deadly human stampedes, information from news reports and the Internet is essential to increased collective understanding. Without incorporating nontraditional sources, no other way to reasonably acquire sufficient data is available. This study analyzed human stampede events from 1980 to 2007 to identify epidemiological characteristics associated with increased mortality. A LexisNexis search was followed by sequential searches of multiple Internet-based English-language news agencies. Date, country, geographical region, time of occurrence, type of event, location, mechanism, number of participants, number injured, and number of deaths were recorded. Bivariate analyses of number of deaths or injuries were conducted using a nonparametric Wilcoxon rank test. Multivariate regression was performed to determine the factors associated with increased number of fatalities during stampede events. A total of 215 human stampede events were reported from 1980 to 2007, resulting in 7069 deaths and at least 14,078 injuries from 213 events with available fatality information and 179 events with injury information. In bivariate analysis, stampedes occurring in the Middle East, in developing countries, outdoors, or associated with religious events had the highest median number of deaths. In multivariate analysis, events that occurred in developing countries and outdoors were associated with increased number of fatalities. Stampedes that occurred in the context of sports, religious, music, and political events, or that had a unidirectional mechanism, also increased the relative number of deaths. Several epidemiological features of human stampedes associated with increased mortality are identified. Standardized collection of epidemiological data pertaining to human stampedes is strongly recommended, and further study of this recurrent, distinctive disaster is warranted.


Elevated plasma interleukin-6 levels in trained male triathletes following an acute period of intense interval training.
The aim of the present study was to investigate possible changes in the plasma IL-6 levels, subjective reporting of sources and symptoms of stress and the innate immune system in response to an acute period of intensified run training in highly trained endurance athletes. Eight healthy endurance trained male subjects (mean +/- SD age 23 +/- 2 years, VO(2max) 64.8 +/- 2.6 ml kg(-1) min(-1), mass 77.1 +/- 2.9 kg) completed the study which took place over a 4 week period. In weeks 2 and 3, in addition to their normal endurance training, subjects completed interval-training run sessions on three successive days. Saliva and venous blood samples were taken at the end of each week. Blood samples were analysed for leukocyte counts; neutrophil function; plasma IL-6; creatine kinase activity; and cortisol. Symptoms and sources of stress were assessed by questionnaire. Plasma IL-6 and creatine kinase activity were elevated following intensified training. Neutrophil function was reduced but total leukocyte and neutrophil counts, plasma cortisol and salivary IgA remained unchanged. There was a worsening in symptoms of stress despite there being no significant change in the sources of stress during intensified training. In conclusion, an acute period of intensified training can induce a suppression of the innate immune system and a chronic elevation in IL-6. This was associated with an increase in fatigue and generalised malaise which lends support to the recent cytokine theories of unexplained, underperformance syndrome.


Glial and neuronal control of brain blood flow.
Blood flow in the brain is regulated by neurons and astrocytes. Knowledge of how these cells control blood flow is crucial for understanding how neural computation is powered, for interpreting functional imaging scans of brains, and for developing treatments for neurological disorders. It is now recognized that neurotransmitter-mediated signalling has a key role in regulating cerebral blood flow, that much of this control is mediated by astrocytes, that oxygen modulates blood flow regulation, and that blood flow may be controlled by capillaries as well as by arterioles. These conceptual shifts in our understanding of cerebral blood flow control have important implications for the development of new therapeutic approaches.


Managing the consequences of aggressive conservative treatment for refractory retinoblastoma with vitreous seeding.
A 4 year-old girl with bilateral, non-familial retinoblastoma (RB) was referred to our care after primary enucleation OS and active tumor OD refractory to multiple therapies (intravenous chemotherapy, laser/cryotherapy, and I-125 plaque radiotherapy). Vitreous seeding OD, initially controlled by several sessions of Ophthalmic Artery Infusion Chemotherapy (OAIC) and periocular chemotherapy, recurred shortly thereafter. The patient underwent intravitreal (IVit) Melphalan injections achieving tumor control despite the concurrent development of keratopathy, pupillary synechiae, cataract, and necrosis of the inferior fornix and the adjacent orbital fat, all secondary to the treatments administered. Repeated amniotic membrane implants and tarsorrhaphy were performed to alleviate the symptoms. Despite being tumor free for 6 months, a poor fundus view and treatment-related complications prompted us to consider enucleation, but parents declined. Following recent negative magnetic resonance imaging (MRI), her cataract was removed. She was then found to have tumor recurrence. Her eye was enucleated 12 months ago and she recovered well from the surgery. As ocular oncology embarks in eye-preserving treatments for retinoblastoma, it is important to address the cumulative effects and associated impact of such treatments and the possibility of failure.


Inhibition of chemical carcinogenesis by viral vaccines.
The incidence of 3-methylcholanthrene-induced subcutaneous tumors was significantly reduced by a single injection of inactivated type C RNA viral vaccine. Rauscher leukemia virus vaccine reduced the incidence of sarcomas from 78 to 50 percent in the BALB/cCr mouse. Radiation leukemia virus vaccine and a vaccine from a wild murine leukemia virus derived from a 3-methylcholanthrene tumor reduced the incidence of sarcoma from 86 percent to 33 and 37 percents, respectively, in the C57BL/6 mouse. These reductions in tumor incidence by virus vaccines help support the concept that type C RNA viruses serve as determinants of chemically induced cancer; additional studies of vaccines made with more purified virus preparations are necessary.


Longitudinal study of the body composition of weight gain in exclusively breast-fed and intake-measured whey-based formula-fed infants to age 3 months.
Body composition and the composition of weight gain in 82 healthy infants during the first 3 months of life were measured in relation to the type of feeding and nutrient intakes by serial measurements of total body potassium (TBK), anthropometry, and formula intake. Infants were exclusively fed either breast milk (n = 34, B fed) or whey-based formula (n = 48, F fed). Formula intakes were substantially lower than WHO-FAO recommendations (although serial weights paralleled standards), and were significantly greater in male infants, who had greater gains in weight and lean tissue than females. In both sexes, tissues low in K (fat and extracellular water) were added faster to the body weight than lean tissue. Overall, compared with B fed infants, F fed infants gained less weight between birth and 10 days and more weight between 10 and 90 days, although mean actual weights at 90 days were similar; formula feeding caused greater fat deposition in males and greater daily gains in lean mass in females. Differences in both quantity and quality of ingested nutrients account for these differences although the biological importance of these findings is speculative. These studies further elucidate the nature of growth in relation to modern feeding practices and the question of optimal versus maximal nutrient intake and growth in early life.


Host genes conferring resistance to a central nervous system disease induced by a polytropic recombinant Friend murine retrovirus.
Infection of certain strains of mice with the ecotropic Friend murine leukemia virus results in the generation of recombinant polytropic mink cell focus-inducing viruses and the development of erythroleukemia. We isolated a Friend mink cell focus-inducing virus (F-MCF-98D) from a Friend murine leukemia virus-infected BALB/c mouse which caused primarily a neurological disease as well as a low incidence of leukemia in susceptible IRW mice. Through genetic studies with the resistant C57BL/10 strain, we identified two genes which correlated with restricted viral replication and resistance to the development of disease caused by F-MCF-98D. One gene correlated with the expression of an endogenous gp70 linked to the Rmcf gene and might act by viral interference. The mechanism of action of the second gene was less clear, but it appeared to be associated with development of an antiviral antibody response.


Genetic determinants of heel bone properties: genome-wide association meta-analysis and replication in the GEFOS/GENOMOS consortium.
Quantitative ultrasound of the heel captures heel bone properties that independently predict fracture risk and, with bone mineral density (BMD) assessed by X-ray (DXA), may be convenient alternatives for evaluating osteoporosis and fracture risk. We performed a meta-analysis of genome-wide association (GWA) studies to assess the genetic determinants of heel broadband ultrasound attenuation (BUA; n = 14 260), velocity of sound (VOS; n = 15 514) and BMD (n = 4566) in 13 discovery cohorts. Independent replication involved seven cohorts with GWA data (in silico n = 11 452) and new genotyping in 15 cohorts (de novo n = 24 902). In combined random effects, meta-analysis of the discovery and replication cohorts, nine single nucleotide polymorphisms (SNPs) had genome-wide significant (P < 5 × 10(-8)) associations with heel bone properties. Alongside SNPs within or near previously identified osteoporosis susceptibility genes including ESR1 (6q25.1: rs4869739, rs3020331, rs2982552), SPTBN1 (2p16.2: rs11898505), RSPO3 (6q22.33: rs7741021), WNT16 (7q31.31: rs2908007), DKK1 (10q21.1: rs7902708) and GPATCH1 (19q13.11: rs10416265), we identified a new locus on chromosome 11q14.2 (rs597319 close to TMEM135, a gene recently linked to osteoblastogenesis and longevity) significantly associated with both BUA and VOS (P < 8.23 × 10(-14)). In meta-analyses involving 25 cohorts with up to 14 985 fracture cases, six of 10 SNPs associated with heel bone properties at P < 5 × 10(-6) also had the expected direction of association with any fracture (P < 0.05), including three SNPs with P < 0.005: 6q22.33 (rs7741021), 7q31.31 (rs2908007) and 10q21.1 (rs7902708). In conclusion, this GWA study reveals the effect of several genes common to central DXA-derived BMD and heel ultrasound/DXA measures and points to a new genetic locus with potential implications for better understanding of osteoporosis pathophysiology.


The influence of ecological and life history factors on ectothermic temperature-size responses: Analysis of three Lycaenidae butterflies (Lepidoptera).
Body size has been shown to decrease with increasing temperature in many species, prompting the suggestion that it is a universal ecological response. However, species with complex life cycles, such as holometabolous insects, may have correspondingly complicated temperature-size responses. Recent research suggests that life history and ecological traits may be important for determining the direction and strength of temperature-size responses. Yet, these factors are rarely included in analyses. Here, we aim to determine whether the size of the bivoltine butterfly, Polyommatus bellargus, and the univoltine butterflies, Plebejus argus and Polyommatus coridon, change in response to temperature and whether these responses differ between the sexes, and for P. bellargus, between generations. Forewing length was measured using digital specimens from the Natural History Museum, London (NHM), from one locality in the UK per species. The data were initially compared to annual and seasonal temperature values, without consideration of life history factors. Sex and generation of the individuals and mean monthly temperatures, which cover the growing period for each species, were then included in analyses. When compared to annual or seasonal temperatures only, size was not related to temperature for P. bellargus and P. argus, but there was a negative relationship between size and temperature for P. coridon. When sex, generation, and monthly temperatures were included, male adult size decreased as temperature increased in the early larval stages, and increased as temperature increased during the late larval stages. Results were similar but less consistent for females, while second generation P. bellargus showed no temperature-size response. In P. coridon, size decreased as temperature increased during the pupal stage. These results highlight the importance of including life history factors, sex, and monthly temperature data when studying temperature-size responses for species with complex life cycles.


Sex-Specific Features of Microglia from Adult Mice.
Sex has a role in the incidence and outcome of neurological illnesses, also influencing the response to treatments. Neuroinflammation is involved in the onset and progression of several neurological diseases, and the fact that estrogens have anti-inflammatory activity suggests that these hormones may be a determinant in the sex-dependent manifestation of brain pathologies. We describe significant differences in the transcriptome of adult male and female microglia, possibly originating from perinatal exposure to sex steroids. Microglia isolated from adult brains maintain the sex-specific features when put in culture or transplanted in the brain of the opposite sex. Female microglia are neuroprotective because they restrict the damage caused by acute focal cerebral ischemia. This study therefore provides insight into a distinct perspective on the mechanisms underscoring a sexual bias in the susceptibility to brain diseases.


Resistance to chytridiomycosis in European plethodontid salamanders of the genus Speleomantes.
North America and the neotropics harbor nearly all species of plethodontid salamanders. In contrast, this family of caudate amphibians is represented in Europe and Asia by two genera, Speleomantes and Karsenia, which are confined to small geographic ranges. Compared to neotropical and North American plethodontids, mortality attributed to chytridiomycosis caused by Batrachochytrium dendrobatidis (Bd) has not been reported for European plethodontids, despite the established presence of Bd in their geographic distribution. We determined the extent to which Bd is present in populations of all eight species of European Speleomantes and show that Bd was undetectable in 921 skin swabs. We then compared the susceptibility of one of these species, Speleomantes strinatii, to experimental infection with a highly virulent isolate of Bd (BdGPL), and compared this to the susceptible species Alytes muletensis. Whereas the inoculated A. muletensis developed increasing Bd-loads over a 4-week period, none of five exposed S. strinatii were colonized by Bd beyond 2 weeks post inoculation. Finally, we determined the extent to which skin secretions of Speleomantes species are capable of killing Bd. Skin secretions of seven Speleomantes species showed pronounced killing activity against Bd over 24 hours. In conclusion, the absence of Bd in Speleomantes combined with resistance to experimental chytridiomycosis and highly efficient skin defenses indicate that the genus Speleomantes is a taxon unlikely to decline due to Bd.