Effects on the nervous system among welders exposed to aluminium and manganese.
The purpose was to study the effects on the nervous system in welders exposed to aluminium and manganese. The investigation included questionnaires on symptoms, psychological methods (simple reaction time, finger tapping speed and endurance, digit span, vocabulary, tracking, symbol digit, cylinders, olfactory threshold, Luria-Nebraska motor scale), neurophysiological methods (electroencephalography, event related auditory evoked potential (P-300), brainstem auditory evoked potential, and diadochokinesometry) and assessments of blood and urine concentrations of metals (aluminium, lead, and manganese). The welders exposed to aluminium (n = 38) reported more symptoms from the central nervous system than the control group (n = 39). They also had a decreased motor function in five tests. The effect was dose related in two of these five tests. The median exposure of aluminium welders was 7065 hours and they had about seven times higher concentrations of aluminium in urine than the controls. The welders exposed to manganese (n = 12) had a decreased motor function in five tests. An increased latency of event related auditory evoked potential was also found in this group. The median manganese exposure was 270 hours. These welders did not have higher concentrations of manganese in blood than the controls. The neurotoxic effects found in the groups of welders exposed to aluminium and manganese are probably caused by the aluminium and manganese exposure, respectively. These effects indicate a need for improvements in the work environments of these welders.


Uterine rupture of an unscarred gravid uterus at term attributed to adenomyosis.
Uterine rupture is a rare obstetric emergency that is typically associated with the presence of scar tissue such as in the case of previous caesarean section. In this case report, a primigravid patient presented to the hospital in cardiac arrest with massive haemoperitoneum secondary to a posterior uterine rupture. The histological specimen was found to have diffuse adenomyosis at the site of rupture. On review of the literature, there is insufficient evidence to suggest we as clinicians should alter the antenatal care for patients with known adenomyosis; however, this case highlights how we should have a high index of suspicion for those presenting with signs and symptoms of uterine rupture with known adenomyosis in the absence of other risk factors.


T(2) -weighted STIR imaging of myocardial edema associated with ischemia-reperfusion injury: the influence of proton density effect on image contrast.
To investigate the contribution of proton density (PD) in T(2) -STIR based edema imaging in the setting of acute myocardial infarction (AMI). Canines (n = 5), subjected to full occlusion of the left anterior descending artery for 3 hours, underwent serial magnetic resonance imaging (MRI) studies 2 hours postreperfusion (day 0) and on day 2. During each study, T(1) and T(2) maps, STIR (TE = 7.1 msec and 64 msec) and late gadolinium enhancement (LGE) images were acquired. Using T(1) and T(2) maps, relaxation and PD contributions to myocardial edema contrast (EC) in STIR images at both TEs were calculated. Edematous territories showed significant increase in PD (20.3 ± 14.3%, P < 0.05) relative to healthy territories. The contributions of T(1) changes and T(2) or PD changes toward EC were in opposite directions. One-tailed t-test confirmed that the mean T(2) and PD-based EC at both TEs were greater than zero. EC from STIR images at TE = 7.1 msec was dominated by PD than T(2) effects (94.3 ± 11.3% vs. 17.6 ± 2.5%, P < 0.05), while at TE = 64 msec, T(2) effects were significantly greater than PD effects (90.8 ± 20.3% vs. 12.5 ± 11.9%, P < 0.05). The contribution from PD in standard STIR acquisitions (TE = 64 msec) was significantly higher than 0 (P < 0.05). In addition to T(2) -weighting, edema detection in the setting of AMI with T(2) -weighted STIR imaging has a substantial contribution from PD changes, likely stemming from increased free-water content within the affected tissue. This suggests that imaging approaches that take advantage of both PD as well as T(2) effects may provide the optimal sensitivity for detecting myocardial edema.


Effects of sustained hyperoxia on revascularization in experimental retinopathy of prematurity.
To investigate the effects of prolonged hyperoxia on vascular recovery and glia survival after experimentally induced retinopathy of prematurity (ROP) in the mouse. The effects of hyperoxia on revascularization and vitreous neovascularization were compared between mice raised in 75% oxygen from postnatal day (P)7 to P12, followed by room air recovery and mice raised in 75% oxygen from P7 to P27. The status of astrocytes and Müller cells was evaluated by glial fibrillary acidic protein (GFAP) immunohistochemistry on retinal wholemounts and serial sections. A window of susceptibility to oxygen-induced vaso-obliteration was defined by comparing the extent of retinal vaso-obliteration resulting from 2 days of hyperoxia beginning on P7, P9, P11, P13, or P15. Oxygen-induced vaso-obliteration of retinal capillaries was limited to the period between birth and P15. Paradoxically, revascularization was markedly accelerated and neovascularization markedly reduced in mice maintained in prolonged hyperoxia (P7-P27) compared with mice recovering in room air. The extended use of 75% oxygen during the recovery period was associated with preservation of astrocytes and Müller cells in the avascular retina. The antiangiogenic effect of hyperoxia on retinal capillaries is strongly dependent on postnatal age. A protocol of continuous 75% supplemental oxygen accelerates recovery of inner retinal vasculature and prevents vitreous neovascularization, by a mechanism that may involve preservation of inner retinal glia.


Coagulopathy associated with poor prognosis in intrahepatic cholangiocarcinoma patients after curative resection.
As a rare type of liver cancer, intrahepatic cholangiocarcinoma (ICC) has become an increasingly important malignancy and continues to present significant therapeutic challenges. Since coagulopathy is associated with poor prognosis in hepatocellular carcinoma (HCC), and prognostic factors of ICC after curative resection were still not clear, we aim to analyze the characteristics of ICC patients with coagulopathy and its correlation to prognosis. From January 2000 to June 2011, 541 ICC patients, after curative resection, were enrolled in our study. Survival curves were depicted by the Kaplan-Meier method and analyzed by the log-rank test. The Cox proportional hazard regression was adopted for multivariate survival analysis. Student's t test was performed to analyze the difference between the coagulopathy group and the normal group. The correlation between coagulation parameters and prognosis was also evaluated. The incidence rate of at least one coagulation parameter abnormality was 22.6% (122/541) while PT was the most common factor (8.87%, 48/541). The one-year survival rate of patients with coagulopathy was significantly lower than that of patients with normal coagulation (p < 0.01). In a univariate analysis, patients with prolonged PT was associated with shortened DFS (p < 0.05). Meanwhile, PT was negatively correlated with pre-albumin level. TNM stage, CA19-9, GGT, and pre-albumin level were independent prognostic factors of DFS in the multivariate analysis. In conclusion, the incidence rate of coagulopathy of ICC patients is lower than HCC patients. Prolonged PT, advanced TNM stage, low pre-albumin level, and high CA19-9 and GGT level were correlated with high recurrence rate and poor prognosis.


ConiferEST: an integrated bioinformatics system for data reprocessing and mining of conifer expressed sequence tags (ESTs).
With the advent of low-cost, high-throughput sequencing, the amount of public domain Expressed Sequence Tag (EST) sequence data available for both model and non-model organism is growing exponentially. While these data are widely used for characterizing various genomes, they also present a serious challenge for data quality control and validation due to their inherent deficiencies, particularly for species without genome sequences. ConiferEST is an integrated system for data reprocessing, visualization and mining of conifer ESTs. In its current release, Build 1.0, it houses 172,229 loblolly pine EST sequence reads, which were obtained from reprocessing raw DNA sequencer traces using our software--WebTraceMiner. The trace files were downloaded from NCBI Trace Archive. ConiferEST provides biologists unique, easy-to-use data visualization and mining tools for a variety of putative sequence features including cloning vector segments, adapter sequences, restriction endonuclease recognition sites, polyA and polyT runs, and their corresponding Phred quality values. Based on these putative features, verified sequence features such as 3' and/or 5' termini of cDNA inserts in either sense or non-sense strand have been identified in-silico. Interestingly, only 30.03% of the designated 3' ESTs were found to have an authenticated 5' terminus in the non-sense strand (i.e., polyT tails), while fewer than 5.34% of the designated 5' ESTs had a verified 5' terminus in the sense strand. Such previously ignored features provide valuable insight for data quality control and validation of error-prone ESTs, as well as the ability to identify novel functional motifs embedded in large EST datasets. We found that "double-termini adapters" were effective indicators of potential EST chimeras. For all sequences with in-silico verified termini/terminus, we used InterProScan to assign protein domain signatures, results of which are available for in-depth exploration using our biologist-friendly web interfaces. ConiferEST represents a unique and complementary public resource for EST data integration and mining in conifers by reprocessing raw DNA traces, identifying putative sequence features and determining and annotating in-silico verified features. Seamlessly integrated with other public resources, ConiferEST provides biologists powerful tools to verify data, visualize abnormalities, including EST chimeras, and explore large EST datasets.


Choroidal Neovascularization Analyzed on Ultrahigh-Speed Swept-Source Optical Coherence Tomography Angiography Compared to Spectral-Domain Optical Coherence Tomography Angiography.
To compare visualization of choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) using an ultrahigh-speed swept-source (SS) optical coherence tomography angiography (OCTA) prototype vs a spectral-domain (SD) OCTA device. Comparative analysis of diagnostic instruments. Patients were prospectively recruited to be imaged on SD OCT and SS OCT devices on the same day. The SD OCT device employed is the RTVue Avanti (Optovue, Inc, Fremont, California, USA), which operates at ∼840 nm wavelength and 70 000 A-scans/second. The SS OCT device used is an ultrahigh-speed long-wavelength prototype that operates at ∼1050 nm wavelength and 400 000 A-scans/second. Two observers independently measured the CNV area on OCTA en face images from the 2 devices. The nonparametric Wilcoxon signed rank test was used to compare area measurements and P values of <.05 were considered statistically significant. Fourteen eyes from 13 patients were enrolled. The CNV in 11 eyes (78.6%) were classified as type 1, 2 eyes (14.3%) as type 2, and 1 eye (7.1%) as mixed type. Total CNV area measured using SS OCT and SD OCT 3 mm × 3 mm OCTA were 0.949 ± 1.168 mm(2) and 0.340 ± 0.301 mm(2), respectively (P = .001). For the 6 mm × 6 mm OCTA the total CNV area using SS OCT and SD OCT were 1.218 ± 1.284 mm(2) and 0.604 ± 0.597 mm(2), respectively (P = .0019). The field of view did not significantly affect the measured CNV area (P = .19 and P = .18 for SS OCT and SD OCT, respectively). SS OCTA yielded significantly larger CNV areas than SD OCTA. It is possible that SS OCTA is better able to demarcate the full extent of CNV vasculature.


Structural basis for drug resistance mechanisms for non-nucleoside inhibitors of HIV reverse transcriptase.
The selection of drug resistant virus is a significant obstacle to the continued successful treatment of HIV infection. Reverse transcriptase is the target for numerous approved anti-HIV drugs including both nucleoside inhibitor (NRTI) and non-nucleosides (NNRTI). The many available crystal structures of RT reveal that, generally, in relation to their binding sites NRTI resistance mutations are generally more distally positioned, whilst for NNRTIs mutations are clustered. Such clustering implies a direct stereochemical basis for NNRTI resistance mechanisms, which is indeed observed in many cases such as the loss of key ring stacking interactions with inhibitors via mutations at Tyr181 and Tyr188. However, there are also indirect resistance mechanisms observed, e.g. V108I (via perturbation of Tyr188 and Tyr181) and K103N (apo-enzyme stabilisation). The resistance mechanism can be NNRTI-dependent as is the case for K101E where either indirect (nevirapine) or direct effects (efavirenz) apply. Structural studies have contributed to the design of newer generation NNRTIs and identified a number of features which may contribute to their much improved resistance profiles. Such factors include reduced interactions with Tyr181, the presence of inhibitor/main-chain H-bonds and ability to undergo conformational flexing and rearrangement within the mutated drug site.


Regional brain electrical activity in posttraumatic stress disorder after motor vehicle accident.
This study examined whether patients with posttraumatic stress disorder (PTSD) related to motor vehicle accidents (MVAs) would show an abnormal pattern of electroencephalographic (EEG) alpha asymmetries, which has been proposed for particular types of anxiety. Patients with PTSD (n = 22) or subsyndromal PTSD (n = 21), traumatized controls without PTSD (non-PTSD with MVA; n = 21), and healthy controls without MVA (n = 23) underwent measurement of EEG activity during baseline and exposure to a neutral, a positive, a negative, and an accident-related picture. Differences in brain asymmetry between groups were observed only during exposure to trauma-related material. PTSD and subsyndromal PTSD patients showed a pattern of enhanced right anterior and posterior activation, whereas non-PTSD with MVA participants showed the opposite pattern. Furthermore, posterior asymmetry in nontraumatized healthy controls varied with gender, with female participants showing a pattern of higher right posterior activation. The results support the hypothesis that symptomatic MVA survivors are characterized by a pattern of right hemisphere activation that is associated with anxious arousal and symptoms of PTSD during processing of trauma-specific information.


A Vision-Based Machine Learning Method for Barrier Access Control Using Vehicle License Plate Authentication.
Automatic vehicle license plate recognition is an essential part of intelligent vehicle access control and monitoring systems. With the increasing number of vehicles, it is important that an effective real-time system for automated license plate recognition is developed. Computer vision techniques are typically used for this task. However, it remains a challenging problem, as both high accuracy and low processing time are required in such a system. Here, we propose a method for license plate recognition that seeks to find a balance between these two requirements. The proposed method consists of two stages: detection and recognition. In the detection stage, the image is processed so that a region of interest is identified. In the recognition stage, features are extracted from the region of interest using the histogram of oriented gradients method. These features are then used to train an artificial neural network to identify characters in the license plate. Experimental results show that the proposed method achieves a high level of accuracy as well as low processing time when compared to existing methods, indicating that it is suitable for real-time applications.


Maternal age influences the number of primordial follicles in the ovaries of yearling Angus heifers.
The number of antral follicles detectable by ultrasonography in heifers is influenced by age of the dam, because daughters of primiparous cows have fewer antral follicles than daughters of mature cows. We, therefore, hypothesized that heifers with primiparous dams would have fewer primordial follicles in their ovaries than heifers born to mature (4+ y) cows. Angus heifers (n = 464) were submitted for ultrasonographic evaluation of antral follicle number at 325, 355, and 385 d of age. Ovaries were collected from a random subset of heifers (n = 79) and processed for histological evaluation to determine number of primordial follicles. A greater percentage of heifers with primiparous dams had a corpus luteum at first ultrasonographic examination; however, a greater percentage of heifers with multiparous dams had ovulated by the start of breeding (P < 0.01). Heifers with primiparous dams had fewer antral follicles detectable by ultrasonography (P < 0.01). Heifers with primparous dams had fewer surface antral follicles on their ovaries (P < 0.01), and the number of primordial follicles per histological section was less for heifers with primiparous dams (P = 0.02). These data indicate that the lesser number of antral follicles detectable by ultrasonography in heifers with primparous dams is due to less ovarian follicle reserves. Selecting replacement heifers from mature dams may result in daughters with greater fertility and reproductive longevity; however, further research is necessary to determine if interactions between size of the ovarian follicle reserve and age at puberty influence fertility and reproductive longevity in replacement heifers.


Complex multimorbidity and health outcomes in older adult cancer survivors.
To characterize complex multimorbidity among cancer survivors and evaluate the association between cancer survivorship, time since cancer diagnosis, and self-reported fair/poor health, self-rated worse health in 2 years, and 2-year mortality. We used the 2010-2012 Health and Retirement Study. Cancer survivors were individuals who reported a (nonskin) cancer diagnosis 2 years or more before the interview. We defined complex multimorbidity as the co-occurrence of chronic conditions, functional limitations, and/or geriatric syndromes. In addition to descriptive analyses, we used logistic regression to evaluate the independent association between cancer survivor status and health outcomes. We also examined whether cancer survivorship differed by the number of years since diagnosis. Among 15,808 older adults (age ≥50 years), 11.8% were cancer survivors. Compared with cancer-free individuals, a greater percentage of cancer survivors had complex multimorbidity: co-occurring chronic conditions, functional limitations, and geriatric syndromes. Cancer survivorship was significantly associated with self-reported fair/poor health, self-rated worse health in 2 years, and 2-year mortality. These effects declined with the number of years since diagnosis for fair/ poor health and mortality but not for self-rated worse health. Cancer survivor status is independently associated with more complex multimorbidity, and with worse health outcomes. These effects attenuate with time, except for patient perception of being in worse health.


Genetic identity, biological phenotype, and evolutionary pathways of transmitted/founder viruses in acute and early HIV-1 infection.
Identification of full-length transmitted HIV-1 genomes could be instrumental in HIV-1 pathogenesis, microbicide, and vaccine research by enabling the direct analysis of those viruses actually responsible for productive clinical infection. We show in 12 acutely infected subjects (9 clade B and 3 clade C) that complete HIV-1 genomes of transmitted/founder viruses can be inferred by single genome amplification and sequencing of plasma virion RNA. This allowed for the molecular cloning and biological analysis of transmitted/founder viruses and a comprehensive genome-wide assessment of the genetic imprint left on the evolving virus quasispecies by a composite of host selection pressures. Transmitted viruses encoded intact canonical genes (gag-pol-vif-vpr-tat-rev-vpu-env-nef) and replicated efficiently in primary human CD4(+) T lymphocytes but much less so in monocyte-derived macrophages. Transmitted viruses were CD4 and CCR5 tropic and demonstrated concealment of coreceptor binding surfaces of the envelope bridging sheet and variable loop 3. 2 mo after infection, transmitted/founder viruses in three subjects were nearly completely replaced by viruses differing at two to five highly selected genomic loci; by 12-20 mo, viruses exhibited concentrated mutations at 17-34 discrete locations. These findings reveal viral properties associated with mucosal HIV-1 transmission and a limited set of rapidly evolving adaptive mutations driven primarily, but not exclusively, by early cytotoxic T cell responses.


Class II histone deacetylases confer signal responsiveness to the ankyrin-repeat proteins ANKRA2 and RFXANK.
Class II histone deacetylases (HDACs) contain unique amino-terminal extensions that mediate interactions with members of the myocyte enhancer factor-2 (MEF2) family of transcription factors and responsiveness to kinases, including Ca2+/calmodulin-dependent kinase (CaMK). Despite intense investigation of class II HDACs, little is known of MEF2-independent mechanisms for transcriptional repression by these chromatin-modifying enzymes. Here, we demonstrate that class II HDACs 4 and 5 physically associate with ankyrin-repeat proteins ANKRA2 and RFXANK (RFX-B/Tvl-1/ANKRA1). ANKRA2 is a megalin- and BKCa potassium channel-interacting factor, whereas RFXANK is a positive regulator of major histocompatibility complex II (MHC II) gene expression. HDAC4 and HDAC5 interact with the ankyrin repeats of ANKRA2 and RFXANK and, through association with RFXANK, repress MHC II promoter activation. HDACs 4 and 5 also repress endogenous HLA-DRA gene expression induced by CIITA. Phosphorylation of class II HDACs by CaMK results in CRM1-dependent nuclear export of HDAC/RFXANK complexes. These results define a novel transcriptional pathway under the control of class II HDACs and suggest a role for these transcriptional repressors as signal-responsive regulators of antigen presentation.


RR interval dynamics before atrial fibrillation in patients after coronary artery bypass graft surgery.
Atrial fibrillation/flutter (AF) is a frequent complication of coronary artery bypass graft surgery (CABG) that leads to increased costs and morbidity. We hypothesized that heart rate variability (HRV), an indicator of cardiac sympathovagal balance, is altered before the onset of postoperative AF. Because nonlinear methods of HRV analysis provide information about heart rate dynamics not evident from usual HRV measures, we also hypothesized that approximate entropy (ApEn), a nonlinear measure of HRV, might have predictive value. Analysis of HRV was performed in 3 sequential 20-minute intervals preceding the onset of postoperative AF (24 episodes in 18 patients). These data were compared with corresponding intervals in 18 sex- and age-matched postoperative control subjects who did not develop AF. Patients had left ventricular ejection fractions >45% before surgery and were not receiving beta-blockers during ambulatory ECG monitoring after surgery. Logistic regression demonstrated that on the basis of averaged values for the three 20-minute intervals, increased heart rate and decreased ApEn were independently associated with AF. Heart rate dynamics before AF was associated with either lower (n= 19) or higher (n=5) RR interval variation by traditional measures of HRV or quantitative Poincaré analysis, suggesting the possibility of divergent autonomic conditions before AF onset. In the hour before AF after CABG surgery, higher heart rate and lower heart rate complexity compared with values in control patients were independent predictors of AF. Decreased ApEn occurs in patients with either increased or decreased HRV by traditional measures and may provide a useful tool for risk stratification or investigation of mechanisms.


Regulation of the cortical actin cytoskeleton in budding yeast by twinfilin, a ubiquitous actin monomer-sequestering protein.
Here we describe the identification of a novel 37-kD actin monomer binding protein in budding yeast. This protein, which we named twinfilin, is composed of two cofilin-like regions. In our sequence database searches we also identified human, mouse, and Caenorhabditis elegans homologues of yeast twinfilin, suggesting that twinfilins form an evolutionarily conserved family of actin-binding proteins. Purified recombinant twinfilin prevents actin filament assembly by forming a 1:1 complex with actin monomers, and inhibits the nucleotide exchange reaction of actin monomers. Despite the sequence homology with the actin filament depolymerizing cofilin/actin-depolymerizing factor (ADF) proteins, our data suggests that twinfilin does not induce actin filament depolymerization. In yeast cells, a green fluorescent protein (GFP)-twinfilin fusion protein localizes primarily to cytoplasm, but also to cortical actin patches. Overexpression of the twinfilin gene (TWF1) results in depolarization of the cortical actin patches. A twf1 null mutation appears to result in increased assembly of cortical actin structures and is synthetically lethal with the yeast cofilin mutant cof1-22, shown previously to cause pronounced reduction in turnover of cortical actin filaments. Taken together, these results demonstrate that twinfilin is a novel, highly conserved actin monomer-sequestering protein involved in regulation of the cortical actin cytoskeleton.


In vivo monitoring of rat brain metabolites during vigabatrin treatment using localized 2D-COSY.
A two-dimensional COSY-based localization sequence was designed to allow the in vivo monitoring of proton metabolites in rat brain [particularly gamma-aminobutyric acid (GABA), glutamine, taurine and myo-inositol]. The sequence incorporated OSIRIS signal localization, B1-insensitive water suppression and phase-sensitive COSY acquisition. The method was used to study the effects of the GABA-transaminase inhibitor vigabatrin on rat brain metabolite concentrations. Wistar rats were treated daily for 3 days with an oral dose of vigabatrin (200 mg/kg, n = 4). Localized COSY spectra were obtained during a 120 min acquisition from a 270 microl central brain voxel and compared with nine untreated control animals. Significant elevations were observed in GABA (267% of control, p < 0.005, Mann-Witney test), glutamine (130% of control, p < 0.005) and taurine (113% of control, p < 0.05). Changes in GABA and taurine were consistent with previous data on the action of Vigabatrin, and support a previously hypothesized link between these compounds. The increase in glutamine was more surprising and may reflect the balance between the level and/or site of GABA-transaminase inhibition and downregulation of GABA synthesis.


Collecting validity evidence for an assessment of professionalism: findings from think-aloud interviews.
This study investigated whether participants' subjective reports of how they assigned ratings on a multisource feedback instrument provide evidence to support interpreting the resulting scores as objective, accurate measures of professional behavior. Twenty-six participants completed think-aloud interviews while rating students, residents, or faculty members they had worked with previously. The items rated included 15 behavioral items and one global item. Participants referred to generalized behaviors and global impressions six times as often as specific behaviors, rated observees in the absence of information necessary to do so, relied on indirect evidence about performance, and varied in how they interpreted items. Behavioral change becomes difficult to address if it is unclear what behaviors raters considered when providing feedback. These findings highlight the importance of explicitly stating and empirically investigating the assumptions that underlie the use of an observational assessment tool.


Spatial regulation of Dia and Myosin-II by RhoGEF2 controls initiation of E-cadherin endocytosis during epithelial morphogenesis.
E-cadherin plays a pivotal role in epithelial morphogenesis. It controls the intercellular adhesion required for tissue cohesion and anchors the actomyosin-driven tension needed to change cell shape. In the early Drosophila embryo, Myosin-II (Myo-II) controls the planar polarized remodelling of cell junctions and tissue extension. The E-cadherin distribution is also planar polarized and complementary to the Myosin-II distribution. Here we show that E-cadherin polarity is controlled by the polarized regulation of clathrin- and dynamin-mediated endocytosis. Blocking E-cadherin endocytosis resulted in cell intercalation defects. We delineate a pathway that controls the initiation of E-cadherin endocytosis through the regulation of AP2 and clathrin coat recruitment by E-cadherin. This requires the concerted action of the formin Diaphanous (Dia) and Myosin-II. Their activity is controlled by the guanine exchange factor RhoGEF2, which is planar polarized and absent in non-intercalating regions. Finally, we provide evidence that Dia and Myo-II control the initiation of E-cadherin endocytosis by regulating the lateral clustering of E-cadherin.


Assessment of the Forward Contamination Risk of Mars by Clean Room Isolates from Space-Craft Assembly Facilities through Aeolian Transport - a Model Study.
The increasing number of missions to Mars also increases the risk of forward contamination. Consequently there is a need for effective protocols to ensure efficient protection of the Martian environment against terrestrial microbiota. Despite the fact of constructing sophisticated clean rooms for spacecraft assembly a 100 % avoidance of contamination appears to be impossible. Recent surveys of these facilities have identified a significant number of microbes belonging to a variety of taxonomic groups that survive the harsh conditions of clean rooms. These microbes may have a strong contamination potential, which needs to be investigate to apply efficient decontamination treatments. In this study we propose a series of tests to evaluate the potential of clean room contaminants to survive the different steps involved in forward contamination. We used Staphylococcus xylosus as model organism to illustrate the different types of stress that potential contaminants will be subjected to on their way from the spacecraft onto the surface of Mars. Staphylococcus xylosus is associated with human skin and commonly found in clean rooms and could therefore contaminate the spacecraft as a result of human activity during the assembling process. The path the cell will take from the surface of the spacecraft onto the surface of Mars was split into steps representing different stresses that include desiccation, freezing, aeolian transport in a Martian-like atmosphere at Martian atmospheric pressure, and UV radiation climate. We assessed the surviving fraction of the cellular population after each step by determining the integrated metabolic activity of the survivor population by measuring their oxygen consumption rate. The largest fraction of the starting culture (around 70 %) was killed during desiccation, while freezing, Martian vacuum and short-term UV radiation only had a minor additional effect on the survivability of Staphylococcus xylosus. The study also included a simulation of atmospheric transport on Martian dust, which did not significantly alter the metabolic potential of the cells. The high survival potential of skin microbes, which are not among the most robust isolates, clearly underlines the necessity for efficient decontamination protocols and of adequate planetary protection measures. Thus we propose a series of tests to be included into the description of isolates from spacecraft assembly clean rooms in order to assess the forward contamination potential of the specific isolate and to categorize the risk level according to the organisms survival potential. We are aware that the tests that we propose do not exhaust the types of challenges that the microbes would meet on their way and therefore the series of tests is open to being extended.


Role of radioactive iodine for adjuvant therapy and treatment of metastases.
Normal thyrocytes and thyroid cancer cells are characterized by possession of a sodium iodide symporter. Radioiodine administration is a unique and powerful means of treating differentiated thyroid cancer because of the ability of thyroid cancer cells to concentrate beta-emitting radiolabeled iodine. Several manipulations, such as iodine depletion and thyroid hormone-stimulating hormone elevation, are used to enhance uptake of radiolabeled iodine by tumor cells. Adjuvant radioiodine therapy, given to patients without evidence of residual disease, enhances the sensitivity of subsequent surveillance and may decrease recurrence rates and mortality. However, its exact role in the management of low-risk patients merits further investigation. In contrast, radioactive iodine therapy used in patients with residual or metastatic disease clearly improves outcomes. Several studies show decreased recurrence and mortality rates in patients treated with radioiodine compared with those not receiving radioactive iodine. Adverse events from radioiodine therapy include salivary gland dysfunction, bone marrow suppression, and reproductive disturbances. Side effects of radioiodine therapy are generally greater when higher activities of radioiodine are used and may be transient or permanent. Secondary malignancies also may occur after radioiodine therapy. These side effects must be weighed against potential benefits, especially when radioactive iodine is used as adjuvant therapy. Stimulation of the expression of the sodium iodide symporter, or its introduction de novo into nonthyroid cells, is promising in treating poorly differentiated thyroid cancer and nonthyroid malignancies, respectively.


Gene expression profile in response to Xanthomonas axonopodis pv. manihotis infection in cassava using a cDNA microarray.
A cassava cDNA microarray based on a large cassava EST database was constructed and used to study the incompatible interaction between cassava and Xanthomonas axonopodis pv. manihotis (Xam) strain CIO151. For microarray construction, 5700 clones from the cassava unigene set were amplified by polymerase chain reaction (PCR) and printed on glass slides. Microarray hybridization was performed using cDNA from cassava plants (resistant variety MBra685) collected at 12, 24, 48 h and 7 and 15 days post-infection as treatment and cDNA from mock-inoculated plants as control. A total of 199 genes were found to be differentially expressed (126 up-regulated and 73 down-regulated). A greater proportion of differentially-expressed genes was observed at 7 days after inoculation. Expression profiling and cluster analyses indicate that, in response to inoculation with Xam, cassava induces dozens of genes, including principally those involved in oxidative burst, protein degradation and pathogenesis-related (PR) genes. In contrast, genes encoding proteins that are involved in photosynthesis and metabolism were down regulated. In addition, various other genes encoding proteins with unknown function or showing no similarity to other proteins were also induced. Quantitative real time PCR experiments confirmed the reliability of our microarray data. In addition we showed that some genes are induced more rapidly in the resistant than in the susceptible cultivar.


Nasal vaccination of young rainbow trout (Oncorhynchus mykiss) against infectious hematopoietic necrosis and enteric red mouth disease.
Determining the earliest age at which farmed fish can be successfully vaccinated is a very important question for fish farmers. Nasal vaccines are novel mucosal vaccines that prevent aquatic infectious diseases of finfish. The present study investigates the ontogeny of the olfactory organ of rainbow trout by histology and aims to establish the earliest age for vaccination against infectious hematopoietic necrosis (IHN) and enteric red mouth (ERM) disease using the nasal route. Rainbow trout (Oncorhynchus mykiss) were vaccinated intranasally (I.N) at three different ages: 1050° days (DD) (group A); 450 DD (group B); and 360 DD (group C), or 70, 30 and 24 days post-hatch (dph), respectively. The mean weights of groups A, B and C were 4.69 g, 2.9 g and 2.37 g, respectively. Fish received either a live attenuated IHN virus vaccine, ERM formalin killed bacterin or saline (mock vaccinated). Fish were challenged to the corresponding live pathogen 28 days post-vaccination. IHN vaccine delivery at 360 DD resulted in 40% mortality likely due to residual virulence of the vaccine. No mortality was observed in the ERM nasal delivery groups. Following challenge, very high protection rates against IHN virus were recorded in all three age groups with survivals of 95%, 100% and 97.5% in groups A, B and C, respectively. Survival against ERM was 82.5%, 87.5% and 77.5% in groups A, B and C, respectively. Survival rates did not differ among ages for either vaccine. Our results indicate the feasibility and effectiveness of nasal vaccination as early as 360 DD and vaccination-related mortalities when a live attenuated viral vaccine was used in the youngest fish.


The future of meningitis vaccines.
Without effective vaccines meningitis remains a substantial worldwide threat with major health-care implications. A number of advances have been made in vaccine design and implementation over the last decade, with new vaccine initiatives providing substantial promise for the future reduction of global disease burden.


Miscarriage, stillbirth and neonatal mortality in the extreme preterm birth window of gestation in a limited-resource setting on the Thailand-Myanmar border: A population cohort study.
Background: The WHO definition of stillbirth uses 28 weeks' gestation as the cut-point, but also defines extreme preterm birth as 24 to <28 weeks' gestation. This presents a problem with the gestational limit of miscarriage, and hence reporting of stillbirth, preterm birth and neonatal death. The objective of this study is to provide a synopsis of the outcome of a population cohort of pregnancies on the Thailand-Myanmar border between 24 to <28 weeks' gestation. Methods: Records from the Shoklo Malaria Research Unit Antenatal Clinics were reviewed for pregnancy outcomes in the gestational window of 24 to <28 weeks, and each record, including ultrasounds reports, were reviewed to clarify the pregnancy outcome. Pregnancies where there was evidence of fetal demise prior to 24 weeks were classified as miscarriage; those viable at 24 weeks' gestation and born before 28 weeks were coded as births, and further subdivided into live- and stillbirth. Results: Between 1995 and 2015, in a cohort of 49,931 women, 0.6% (318) of outcomes occurred from 24 to <28 weeks' gestation, and 35.8% (114) were miscarriages, with confirmatory ultrasound of fetal demise in 45.4% (49/108). Of pregnancies not ending in miscarriage, 37.7% (77/204) were stillborn and of those born alive, neonatal mortality was 98.3% (115/117). One infant survived past the first year of life. Congenital abnormality rate was 12.0% (23/191). Ultrasound was associated with a greater proportion of pregnancy outcome being coded as birth. Conclusion: In this limited-resource setting, pregnancy outcome from 24 to <28 weeks' gestation included: 0.6% of all outcomes, of which one-third were miscarriages, one-third of births were stillborn and mortality of livebirths approached 100%. In the scale-up to preventable newborns deaths, at least initially, greater benefits will be obtained by focusing on the greater number of viable newborns with a gestation of 28 weeks or more.


[Agrobacterium tumefaciens mediated transformation of Aspergillus niger conidiospores and sequence analysis of T-DNA insertion site in conidiation-defective mutant].
Development of a method of transforming Aspergillus niger conidiospores with Agrobacterium tumefaciens T-DNA system and using it as a tool for genome annotation via construction a T-DNA insertion mutant library of A. niger. A. tumefaciens EHA105 carrying the binary vector pCAMBIA1301 was used to transform A. niger conidiospores under induction conditions. Hygromycin resistant transformants were screened on the selected medium. Sequence analysis was performed using inverse PCR method in stable mutants. In total, 193 hygromycin resistant stable transformants were obtained and the transformation rate was about 5.6 x 10(2) transformants /10(8) conidiospores. Obvious morphological changes were observed in some mutants and sequence analysis of T-DNA inserted site was performed in one conidiation-defective mutant. The result showed that the interrupted gene belonged to major facilitator superfamily (MFS). Our results indicated that the transformation system of A. niger conidiospores set up in this study, combing with sequence analysis of T-DNA insertion site, might be an effective way for genome annotation in A. niger.


Activation of central melanocortin pathways by fenfluramine.
D-fenfluramine (d-FEN) was once widely prescribed and was among the most effective weight loss drugs, but was withdrawn from clinical use because of reports of cardiac complications in a subset of patients. Discerning the neurobiology underlying the anorexic action of d-FEN may facilitate the development of new drugs to prevent and treat obesity. Through a combination of functional neuroanatomy, feeding, and electrophysiology studies in rodents, we show that d-FEN-induced anorexia requires activation of central nervous system melanocortin pathways. These results provide a mechanistic explanation of d-FEN's anorexic actions and indicate that drugs targeting these downstream melanocortin pathways may prove to be effective and more selective anti-obesity treatments.


Thalamocortical oscillations in the sleeping and aroused brain.
Sleep is characterized by synchronized events in billions of synaptically coupled neurons in thalamocortical systems. The activation of a series of neuromodulatory transmitter systems during awakening blocks low-frequency oscillations, induces fast rhythms, and allows the brain to recover full responsiveness. Analysis of cortical and thalamic networks at many levels, from molecules to single neurons to large neuronal assemblies, with a variety of techniques, ranging from intracellular recordings in vivo and in vitro to computer simulations, is beginning to yield insights into the mechanisms of the generation, modulation, and function of brain oscillations.


Uranium and other contaminants in hair from the parents of children with congenital anomalies in Fallujah, Iraq.
Recent reports have drawn attention to increases in congenital birth anomalies and cancer in Fallujah Iraq blamed on teratogenic, genetic and genomic stress thought to result from depleted Uranium contamination following the battles in the town in 2004. Contamination of the parents of the children and of the environment by Uranium and other elements was investigated using Inductively Coupled Plasma Mass Spectrometry. Hair samples from 25 fathers and mothers of children diagnosed with congenital anomalies were analysed for Uranium and 51 other elements. Mean ages of the parents was: fathers 29.6 (SD 6.2); mothers: 27.3 (SD 6.8). For a sub-group of 6 women, long locks of hair were analysed for Uranium along the length of the hair to obtain information about historic exposures. Samples of soil and water were also analysed and Uranium isotope ratios determined. Levels of Ca, Mg, Co, Fe, Mn, V, Zn, Sr, Al, Ba, Bi, Ga, Pb, Hg, Pd and U (for mothers only) were significantly higher than published mean levels in an uncontaminated population in Sweden. In high excess were Ca, Mg, Sr, Al, Bi and Hg. Of these only Hg can be considered as a possible cause of congenital anomaly. Mean levels for Uranium were 0.16 ppm (SD: 0.11) range 0.02 to 0.4, higher in mothers (0.18 ppm SD 0.09) than fathers (0.11 ppm; SD 0.13). The highly unusual non-normal Fallujah distribution mean was significantly higher than literature results for a control population Southern Israel (0.062 ppm) and a non-parametric test (Mann Whitney-Wilcoxon) gave p = 0.016 for this comparison of the distribution. Mean levels in Fallujah were also much higher than the mean of measurements reported from Japan, Brazil, Sweden and Slovenia (0.04 ppm SD 0.02). Soil samples show low concentrations with a mean of 0.76 ppm (SD 0.42) and range 0.1-1.5 ppm; (N = 18). However it may be consistent with levels in drinking water (2.28 μgL-1) which had similar levels to water from wells (2.72 μgL-1) and the river Euphrates (2.24 μgL-1). In a separate study of a sub group of mothers with long hair to investigate historic Uranium excretion the results suggested that levels were much higher in the past. Uranium traces detected in the soil samples and the hair showed slightly enriched isotopic signatures for hair U238/U235 = (135.16 SD 1.45) compared with the natural ratio of 137.88. Soil sample Uranium isotope ratios were determined after extraction and concentration of the Uranium by ion exchange. Results showed statistically significant presence of enriched Uranium with a mean of 129 with SD5.9 (for this determination, the natural Uranium 95% CI was 132.1 < Ratio < 144.1). Whilst caution must be exercised about ruling out other possibilities, because none of the elements found in excess are reported to cause congenital diseases and cancer except Uranium, these findings suggest the enriched Uranium exposure is either a primary cause or related to the cause of the congenital anomaly and cancer increases. Questions are thus raised about the characteristics and composition of weapons now being deployed in modern battlefields.


Multimorbidity and mortality in older adults: A systematic review and meta-analysis.
To review literature and provide a pooled effect for the association between multimorbidity and mortality in older adults. A systematic review was performed of articles held on the PUBMED database published up until January 2015. Studies which used different diseases and other conditions to define frailty, evaluated multimorbidity related only to mental health or which presented disease homogeneity were not included. A meta-analysis using random effect to obtain a pooled effect of multimorbidity on mortality in older adults was conducted only with studies which reported hazard ratio (HR). Stratified analysis and univariate meta-regression were performed to evaluate sources of heterogeneity. Out of 5806 identified articles, 26 were included in meta-analysis. Overall, positive association between multimorbidity and mortality [HR: 1.44 (95%CI: 1.34; 1.55)] was detected. The number of morbidities was positively related to risk of death [HR: 1.20 (95%CI: 1.10; 1.30)]. Compared to individuals without multimorbidity, the risk of death was 1.73 (95%CI: 1.41; 2.13) and 2.72 (95%CI: 1.81; 4.08) for people with 2 or more and 3 or more morbidities, respectively. Heterogeneity between studies was high (96.5%). The sample, adjustment and follow-up modified the associations. Only nine estimates performed adjustment which included demographic, socioeconomic and behaviour variables. Disabilities appear to mediate the effect of multimorbidity on mortality. Multimorbidity was associated with an increase in risk of death. Multimorbidity measurement standardization is needed to produce more comparable estimates. Adjusted analysis which includes potential confounders might contribute to better understanding of causal relationships between multimorbidity and mortality.


Metals in feathers of black-crowned night-heron (Nycticorax nycticorax) chicks from the New York Harbor Estuary.
In heavily urbanized landscapes such as the New York Harbor Estuary, the local environment is subject to a variety of contamination sources. Environmental contaminants such as heavy metals and metalloids pose a risk to wildlife inhabiting the harbor. Metal concentrations in feathers indicate exposure and provide insight into the potential adverse effects on birds. In 2004 and 2005 down feathers were collected from 147 black-crowned night heron (Nycticorax nycticorax) chicks living on four island colonies in the New York Harbor Estuary, USA: Goose Island, Hoffman Island, North Brother Island, and Canarsie Pol. We examined geographical and interannual differences in concentrations of the metalloid arsenic (As) and the heavy metals cadmium (Cd), chromium (Cr), lead (Pb), and mercury (Hg). Significant differences in metal concentrations were a function of location, with North Brother Island having the highest As concentrations and Goose Island having the highest Cr and Pb concentrations in feathers collected in 2004. In 2005, feather samples indicated Hoffman Island had the highest Cr concentrations and North Brother Island had the highest Pb and Hg concentrations. Concentrations of As, Cd, and Hg decreased significantly from 2004 to 2005 on North Brother Island, Hg on Hoffman Island decreased from 2004 to 2005, while Cr on Hoffman Island increased from 2004 to 2005. Cd and Pb concentrations were higher and As and Cr concentrations were generally lower than concentrations reported in previous studies. Further investigation is necessary on the declining population of herons in the New York Harbor Estuary to determine specifically if birds are experiencing adverse effects from metal contaminants, or if organic contaminants, such as organic Hg or polychlorinated biphenyls (PCBs), are playing a role in this population's decline.


Fluid dynamics of flapping aquatic flight in the bird wrasse: three-dimensional unsteady computations with fin deformation.
Many fishes that swim with the paired pectoral fins use fin-stroke parameters that produce thrust force from lift in a mechanism of underwater flight. These locomotor mechanisms are of interest to behavioral biologists, biomechanics researchers and engineers. In the present study, we performed the first three-dimensional unsteady computations of fish swimming with oscillating and deforming fins. The objective of these computations was to investigate the fluid dynamics of force production associated with the flapping aquatic flight of the bird wrasse Gomphosus varius. For this computational work, we used the geometry of the wrasse and its pectoral fin, and previously measured fin kinematics, as the starting points for computational investigation of three-dimensional (3-D) unsteady fluid dynamics. We performed a 3-D steady computation and a complete set of 3-D quasisteady computations for a range of pectoral fin positions and surface velocities. An unstructured, grid-based, unsteady Navier-Stokes solver with automatic adaptive remeshing was then used to compute the unsteady flow about the wrasse through several complete cycles of pectoral fin oscillation. The shape deformation of the pectoral fin throughout the oscillation was taken from the experimental kinematics. The pressure distribution on the body of the bird wrasse and its pectoral fins was computed and integrated to give body and fin forces which were decomposed into lift and thrust. The velocity field variation on the surface of the wrasse body, on the pectoral fins and in the near-wake was computed throughout the swimming cycle. We compared our computational results for the steady, quasi-steady and unsteady cases with the experimental data on axial and vertical acceleration obtained from the pectoral fin kinematics experiments. These comparisons show that steady state computations are incapable of describing the fluid dynamics of flapping fins. Quasi-steady state computations, with correct incorporation of the experimental kinematics, are useful when determining trends in force production, but do not provide accurate estimates of the magnitudes of the forces produced. By contrast, unsteady computations about the deforming pectoral fins using experimentally measured fin kinematics were found to give excellent agreement, both in the time history of force production throughout the flapping strokes and in the magnitudes of the generated forces.


Self-assembled glycol chitosan nanoparticles for disease-specific theranostics.
Hydrophobically modified glycol chitosan (hGC) conjugates spontaneously form self-assembled nanoparticles (NPs) in aqueous conditions, and glycol chitosan NPs (CNPs) have been extensively studied for the past few decades. For disease-specific theranostics, CNPs could be simply modified with imaging agents, and the hydrophobic domains of hGC are available for encapsulation of various drugs. Based on the excellent physiochemical and biological properties, CNPs have been investigated for multimodal imaging and target specific drug delivery. In particular, a recent application of CNPs has shown great potential as an efficient theranostic system because the CNPs could be utilized for a disease-specific theranostic delivery system of different imaging agents and therapeutics, simultaneously. Furthermore, various therapeutic agents including chemo-drugs, nucleotides, peptides, and photodynamic chemicals could be simply encapsulated into the CNPs through hydrophobic or charge-charge interactions. Under in vivo conditions, the encapsulated imaging agents and therapeutic drugs have been successfully delivered to targeted diseases. In this article, the overall research progress on CNPs is reviewed from early works. The current challenges of CNPs to overcome in theranostics are also discussed, and continuous studies would provide more opportunities for early diagnosis of diseases and personalized medicine.


Digoxin and Hypermagnesuria.
In a recent issue of Nephron, Abu-Amer et al.[<xref ref-type="bibr" rid="ref1">1</xref>] reported the presence of hypermagnesuria in patients following acute intravenous administration of digoxin and suggested that the Na+/K+-ATPase γ-subunit, which is the pharmacological target of digoxin, can play a role in this process. Hypermagnesuria induced by digoxin may have important clinical consequences, particularly in the presence of inherited and acquired conditions associated with hypermagnesuria and hypomagnesemia. Moreover, the co-administration of digoxin with other drugs that reduce gastrointestinal absorption (i.e., proton pump inhibitors) or increase urinary excretion (i.e., loop diuretics) may increase the likelihood of developing hypomagnesemia. In this article, we reviewed the main causes of hypermagnesuria and discussed potential drug interactions that can enhance the magnesuric effect of digoxin. We suggest that during the administration of digoxin, clinicians should consider the presence of other causes of hypomagnesemia and hypermagnesuria that could enhance the magnesuric effect of digoxin, monitor the urinary and serum levels of magnesium and prescribe an oral supplementation of magnesium.


ADGRL4/ELTD1 is a highly conserved angiogenesis-associated orphan adhesion GPCR that emerged with the first vertebrates and comprises 3 evolutionary variants.
Our laboratory identified ADGRL4/ELTD1, an orphan GPCR belonging to the adhesion GPCR (aGPCR) family, as a novel regulator of angiogenesis and a potential anti-cancer therapeutic target. Little is known about how ADGRL4/ELTD1 (and aGPCRs in general) function, a problem compounded by a lack of known ligands or means of activation. With this in mind, we turned to computational evolutionary biology with the aim of better understanding ADGRL4/ELTD1. We identified ADGRL4/ELTD1 as a highly conserved early angiogenic gene which emerged in the first true vertebrates (bony fish) approximately 435 million years ago (mya), evolving alongside key angiogenic genes VEGFR2 and DLL4. We identified 3 evolutionary ADGRL4/ELTD1 variants based on EGF domain deletions with variant 2 first emerging 101 mya (95% CI 96-105) in Afrotheria and 82 mya (95% CI 76-89) in Primates. Additionally, conservation mapping across all orthologues reveals highest level conservation in EGF Ca binding domain 1, suggesting that this motif plays an essential role, as well as specific regions of the GAIN domain, GPS motif and 7TM domain, suggesting possible activation mechanisms and ligand binding positions. Additionally, we found that ADGRL4/ELTD1 (a member aGPCR family 1) is possibly ancestral to members of aGPCR family 2. This work establishes ADGRL4/ELTD1's evolution, sheds light on its possible activation and ligand binding zones, and establishes the first temporal references for the emergence of ADGRL4/ELTD1 variants during vertebrate evolution. Our approach is applicable to the greater aGPCR family and opens up new avenues for future experimental work.


Prevalence of intimate partner violence and associated factors amongst women attending antenatal care at Outapi clinic, Namibia: A descriptive survey.
Intimate partner violence (IPV) is a significant and largely hidden public health problem for all women and, during pregnancy, can have significant effects on the health of both mother and the unborn baby. Previous Namibian studies suggest rates of IPV as high as 36%, although few studies have been conducted in primary care. To determine the prevalence of IPV amongst women attending antenatal care. Outapi primary care clinic, Namibia. A descriptive survey administering a validated questionnaire to 386 consecutive participants. The mean age of the participants was 27.5 years (standard deviation = 6.8), 335 (86.8%) were unmarried, 215 (55.7%) had only primary school education and 237 (61.4%) were in their third trimester. Overall, 51 participants (13.2%) had HIV and 44 (11.4%) had teenage pregnancies. The reported lifetime prevalence of IPV was 39 (10.1%), the 12-month prevalence was 35 (9.1%) and the prevalence during pregnancy was 31 (8.0%). Emotional abuse was the commonest type of abuse in 27 (7.0%). The commonest specific abusive behaviour was refusing to provide money to run the house or look after the children whilst the partner spent money on his priorities (4.9%). Increased maternal age was associated with an increase in the occurrence of IPV. The reported lifetime prevalence of IPV was 10.1%, with emotional abuse being the commonest type of abuse. Increased age was associated with an increase in reported IPV. IPV is significant enough to warrant that healthcare providers develop guidelines to assist women affected by IPV in Namibia.


Corticotropin-releasing factor and urocortin regulate spine and synapse formation: structural basis for stress-induced neuronal remodeling and pathology.
Dendritic spines are important sites of excitatory neurotransmission in the brain with their function determined by their structure and molecular content. Alterations in spine number, morphology and receptor content are a hallmark of many psychiatric disorders, most notably those because of stress. We investigated the role of corticotropin-releasing factor (CRF) stress peptides on the plasticity of spines in the cerebellum, a structure implicated in a host of mental illnesses, particularly of a developmental origin. We used organotypic slice cultures of the cerebellum and restraint stress in behaving animals to determine whether CRF in vitro and stress in vivo affects Purkinje cell (PC) spine density. Application of CRF and urocortin (UCN) to cerebellar slice cultures increased the density of spines on PC signaling via CRF receptors (CRF-Rs) 1 and 2 and RhoA downregulation, although the structural phenotypes of the induced spines varied, suggesting that CRF-Rs differentially induce the outgrowth of functionally distinct populations of spines. Furthermore, CRF and UCN exert a trophic effect on the surface contact between synaptic elements by increasing active zones and postsynaptic densities and facilitating the alignment of pre- and post-synaptic membranes of synapses on PCs. In addition, 1 h of restraint stress significantly increased PC spine density compared with those animals that were only handled. This study provides unprecedented resolution of CRF pathways that regulate the structural machinery essential for synaptic transmission and provides a basis for understanding stress-induced mental illnesses.


Social stress induces neurovascular pathology promoting depression.
Studies suggest that heightened peripheral inflammation contributes to the pathogenesis of major depressive disorder. We investigated the effect of chronic social defeat stress, a mouse model of depression, on blood-brain barrier (BBB) permeability and infiltration of peripheral immune signals. We found reduced expression of the endothelial cell tight junction protein claudin-5 (Cldn5) and abnormal blood vessel morphology in nucleus accumbens (NAc) of stress-susceptible but not resilient mice. CLDN5 expression was also decreased in NAc of depressed patients. Cldn5 downregulation was sufficient to induce depression-like behaviors following subthreshold social stress whereas chronic antidepressant treatment rescued Cldn5 loss and promoted resilience. Reduced BBB integrity in NAc of stress-susceptible or mice injected with adeno-associated virus expressing shRNA against Cldn5 caused infiltration of the peripheral cytokine interleukin-6 (IL-6) into brain parenchyma and subsequent expression of depression-like behaviors. These findings suggest that chronic social stress alters BBB integrity through loss of tight junction protein Cldn5, promoting peripheral IL-6 passage across the BBB and depression.


Rescue of a severely bottlenecked wolf (Canis lupus) population by a single immigrant.
The fragmentation of populations is an increasingly important problem in the conservation of endangered species. Under these conditions, rare migration events may have important effects for the rescue of small and inbred populations. However, the relevance of such migration events to genetically depauperate natural populations is not supported by empirical data. We show here that the genetic diversity of the severely bottlenecked and geographically isolated Scandinavian population of grey wolves (Canis lupus), founded by only two individuals, was recovered by the arrival of a single immigrant. Before the arrival of this immigrant, for several generations the population comprised only a single breeding pack, necessarily involving matings between close relatives and resulting in a subsequent decline in individual heterozygosity. With the arrival of just a single immigrant, there is evidence of increased heterozygosity, significant outbreeding (inbreeding avoidance), a rapid spread of new alleles and exponential population growth. Our results imply that even rare interpopulation migration can lead to the rescue and recovery of isolated and endangered natural populations.


The rluC gene of Escherichia coli codes for a pseudouridine synthase that is solely responsible for synthesis of pseudouridine at positions 955, 2504, and 2580 in 23 S ribosomal RNA.
Escherichia coli ribosomal RNA contains 10 pseudouridines, one in the 16 S RNA and nine in the 23 S RNA. Previously, the gene for the synthase responsible for the 16 S RNA pseudouridine was identified and cloned, as was a gene for a synthase that makes a single pseudouridine in 23 S RNA. The yceC open reading frame of E. coli is one of a set of genes homologous to these previously identified ribosomal RNA pseudouridine synthases. In this work, the gene was cloned, overexpressed, and shown to code for a pseudouridine synthase able to react with in vitro transcripts of 23 S ribosomal RNA. Deletion of the gene and analysis of the 23 S RNA from the deletion strain for the presence of pseudouridine at its nine known sites revealed that this synthase is solely responsible in vivo for the synthesis of three of the nine pseudouridine residues, at positions 955, 2504, and 2580. Therefore, this gene has been renamed rluC. Despite the absence of one-third of the normal complement of pseudouridines, there was no change in the exponential growth rate in either LB or M-9 medium at temperatures ranging from 24 to 42 degrees C. From this work and our previous studies, we have now identified three synthases that account for 50% of the pseudouridines in the E. coli ribosome.


Inflammation up-regulates cochlear expression of TRPV1 to potentiate drug-induced hearing loss.
Aminoglycoside antibiotics are essential for treating life-threatening bacterial infections, despite the risk of lifelong hearing loss. Infections induce inflammation and up-regulate expression of candidate aminoglycoside-permeant cation channels, including transient receptor potential vanilloid-1 (TRPV1). Heterologous expression of TRPV1 facilitated cellular uptake of (fluorescently tagged) gentamicin that was enhanced by agonists, and diminished by antagonists, of TRPV1. Cochlear TRPV1 was immunolocalized near the apical membranes of sensory hair cells, adjacent supporting cells, and marginal cells in the stria vascularis. Exposure to immunostimulatory lipopolysaccharides, to simulate of bacterial infections, increased cochlear expression of TRPV1 and hair cell uptake of gentamicin. Lipopolysaccharide exposure exacerbated aminoglycoside-induced auditory threshold shifts and loss of cochlear hair cells in wild-type, but not in heterozygous Trpv1+/- or Trpv1 knockout, mice. Thus, TRPV1 facilitates cochlear uptake of aminoglycosides, and bacteriogenic stimulation upregulates TRPV1 expression to exacerbate cochleotoxicity. Furthermore, loss-of-function polymorphisms in Trpv1 can protect against immunogenic exacerbation of aminoglycoside-induced cochleotoxicity.


Purification and characterization of three extracellular (1-->3)-beta-D-glucan glucohydrolases from the filamentous fungus Acremonium persicinum.
Three (1-->3)-beta-D-glucanases (GNs) were isolated from the culture filtrates of the filamentous fungus Acremonium persicinum and purified by (NH4)2SO4 precipitation followed by anion-exchange and gel-filtration chromatography. Homogeneity of the purified proteins was confirmed by SDS/PAGE, isoelectric focusing and N-terminal amino acid sequencing. All three GNs (GN I, II and III) are non-glycosylated, monomeric proteins with apparent molecular masses, estimated by SDS/PAGE, of 81, 85 and 89 kDa respectively. pI values for the three enzymes are 5.3, 5.1, and 4.4 respectively. The pH optimum for GN I is 6.5, and 5.0 for GN II and III. All three purified enzymes displayed stability over the pH range 4.5-10.0. Optimum activities for GN I, II and III were recorded at 65, 55 and 60 degrees C respectively, with both GN II and III having short-term stability up to 50 degrees C and GN I up to 55 degrees C. The purified GNs have high specificity for (1-->3)-beta-linkages and hydrolysed a range of (1-->3)-beta- and (1-->3)(1-->6)-beta-D-glucans, with laminarin from Laminaria digitata being the most rapidly hydrolysed substrate of those tested. K(m) values for GN I, II, and III against L. digitata laminarin were 0.1, 0.23 and 0.22 mg/ml respectively. D-Glucono-1,5-lactone does not inhibit any of the three GNs, some metals ions are mild inhibitors, and N-bromosuccinimide and KMnO4 are strong inhibitors. All three GNs acted in an exo-hydrolytic manner, determined by the release of alpha-glucose as the initial and major product of hydrolysis of (1-->3)-beta-D-glucans, and confirmed by viscometric analysis and the inability to cleave periodate-oxidized laminarin, and may be classified as (1-->3)-beta-D-glucan glucohydrolases (EC 188.8.131.52).


New directions in the treatment of infected pleural effusion.
The optimum management of patients with parapneumonic effusion and empyema remains uncertain. This article will review the evidence and current opinion on the pathophysiology of this disease, the role of fibrinolytic therapy, and the use of modern surgical techniques.


Cell cycle-dependent complex formation of BRCA1.CtIP.MRN is important for DNA double-strand break repair.
BRCA1 plays an important role in the homologous recombination (HR)-mediated DNA double-strand break (DSB) repair, but the mechanism is not clear. Here we describe that BRCA1 forms a complex with CtIP and MRN (Mre11/Rad50/Nbs1) in a cell cycle-dependent manner. Significantly, the complex formation, especially the ionizing radiation-enhanced association of BRCA1 with MRN, requires cyclin-dependent kinase activity. CtIP directly interacts with Nbs1. The in vivo association of BRCA1 with MRN is largely dependent on the association of CtIP with the BRCT domains at the C terminus of BRCA1, whereas the N terminus of BRCA1 also contributes to its association with MRN. CtIP, as well as the interaction of BRCA1 with CtIP and MRN, is critical for IR-induced single-stranded DNA formation and cellular resistance to radiation. Consistently, CtIP itself is required for efficient HR-mediated DSB repair, like BRCA1 and MRN. These studies suggest that the complex formation of BRCA1.CtIP.MRN is important for facilitating DSB resection to generate single-stranded DNA that is needed for HR-mediated DSB repair. Because cyclin-dependent kinase is important for establishing IR-enhanced interaction of MRN with BRCA1, we propose that the cell cycle-dependent complex formation of BRCA1, CtIP, and MRN contributes to the activation of HR-mediated DSB repair in the S and G(2) phases of the cell cycle.


European brown hare syndrome virus in free-ranging European brown hares from Argentina.
From 1998 to 2000, serum samples of 80 shot European brown hares (Lepus europaeus) from Argentina were examined for antibodies against European brown hare syndrome virus (EBHSV) and 80 spleen samples were tested for EBHSV-antigen by enzyme linked immunosorbent assay (ELISA). Nine hares were positive for EBHSV-antigen. Antibodies against EBHSV were detected in only one individual. Based on negative staining electron microscopy of spleen homogenates, we observed calicivirus in one of five EBHSV-antigen positive hares. However, EBHS has not been reported to cause abnormal mortality in these hares. This is the first report of antibodies to EBHSV, EBHSV-antigen, and electron microscopy findings in free-ranging European brown hares from South America.


Ultrastructural observations on microgametogenesis and the structure of the microgamete of Isospora felis.
The endogenous forms of Isospora felis were observed within the epithelial cells of the small intestine of the cat. They were situated within a parasitophorous vacuole which was limited by a multimembranous wall. The ultrastructural features of microgametogenesis were studied at 8 days post-infection. The initial phase of microgamont development consisted of cytoplasmic growth accompanied by a number of nuclear divisions. The gamont was enclosed by a pellicle and its surface area was greatly increased by deep invaginations. In the later stages of development the numerous nuclei were situated close to the pellicle. Each nucleus has peripherally condensed chromatin. Formation of the microgametes occurred as protrusions from the microgamont surface. Two basal bodies, the dense portion of a nucleus and a mitochondrion entered each protrusion. The microgametes matured while still attached to the gamont from which they finally budded off into the parasitophorous vacuole leaving a large residual cytoplasmic mass. The mature microgamete was found to consist of an elogate nucleus which overlaps with a mitochondrion towards the anterior end of the organism. The anterior portion contains a dense perforatorium and two basal bodies with attached flagella. In addition a number of microtubules (5-9) were found to run longitudinally from the basal body region.


Multi-use of the sea: A wide array of opportunities from site-specific cases across Europe.
The concept of multi-use of the sea has gained popularity in recent years as a result of ocean space (coastal areas and regions with relatively small sea space in particular) becoming increasingly crowded due to the development of the maritime economy. Competing claims for space can be a source of conflict, however this may also lead to mutual benefits for different users when sustainable combinations are sought. Despite increasing European-wide efforts, on-the-ground knowledge and practice of multi-use are still limited. Therefore, with the aim of investigating opportunities for multi-use development in the European seas, 10 case studies were selected, involving different site-specific contexts. This study analyses the characteristics and development potential for ocean multi-use, integrating results from desk analysis and stakeholder perceptions from different sectors in each of the case study locations. Similarities and differences between various combinations of sea uses are also identified. The results show a high heterogeneity of multi-use opportunities between case studies, with a range of combinations identified. The investigated combinations of maritime uses share an overall balance between factors promoting (drivers) and hindering (barriers) multi-use development. Based on stakeholder opinions, expected benefits (added values) of multi-use implementation outweigh potential negative impacts. Management actions are also proposed to further exploit multi-use potential at a local, regional (sub-national) and national levels.


Catalytic mechanism of fungal homoserine transacetylase.
Homoserine transacetylase is a required catalyst in the biochemical pathway that metabolizes Asp to Met in fungi. The enzyme from the yeast Schizosaccharomyces pombe activates the hydroxyl group of L-homoserine by acetylation from acetyl coenzyme A. This enzyme is unique to fungi and some bacteria and presents an important new target for drug discovery. Steady-state kinetic parameters provide evidence that this enzyme follows a ping-pong mechanism. Proton inventory was consistent with a single-proton transfer, and pH studies suggested the participation of at least one residue with a pKa value of 6.4-6.6, possibly a His or Asp/Glu in catalysis. Protein sequence alignments indicate that this enzyme belongs to the alpha/beta-hydrolase fold superfamily of enzymes, indicating the involvement of an active-site nucleophile and possibly a canonical catalytic triad. We constructed site-specific mutants and identified Ser163, Asp403, and His432 as the likely active-site residues of a catalytic triad based on steady-state kinetics and genetic complementation of a yeast null mutant. Moreover, unlike the wild-type enzyme, inactive site mutants were not capable of producing an acetyl-enzyme intermediate. Homoserine transacetylase therefore catalyzes the acetylation of L-homoserine via a covalent acyl-enzyme intermediate through an active-site Ser. These results form the basis of future exploitation of this enzyme as an antimicrobial target.


Effects of esterification, saturation and amount of fatty acids infused into the rumen or abomasum in lactating dairy cows.
Our objective was to determine the effects of chemical structure, amount, and site of infusion of long-chain fatty acids (LCFA) in lactating dairy cows. Six multiparous Holstein cows were used in a 6 × 6 Latin square design with 21-d periods. During d 1 to 14, 250 g/d of LCFA and during d 15 to 21, 500 g/d of LCFA were infused continuously into either the rumen or abomasum. Treatments were 1) Control (CONT); 200 g/d of meat solubles plus 12 g/d of Tween 80 in 10 L of water, administered half in the rumen and half in abomasum; 2) control plus mostly saturated LCFA into the abomasum (SFAA); 3) control plus mostly saturated LCFA into the rumen (SFAR); 4) control plus soy (mostly unsaturated LCFA) free fatty acids (FFA) into the abomasum (UFAA); 5) control plus soy triglycerides (TG) into the abomasum (TGA); and 6) control plus soy TG into the rumen (TGR). The first 10 d of each period were for adaptation and washout from the previous treatment. The diet consisted of 30% (dry matter basis) corn silage, 20% alfalfa silage and 50% concentrate. Cows infused with UFAA had lower dry matter intake and milk yield than those infused with SFAA or TGA and reductions were greater at the higher infusion amount. Milk fat yield was decreased by UFAA relative to other treatments. Unsaturated LCFA decreased milk fat yield more than saturated LCFA. All LCFA treatments decreased short- and medium-chain FA in milk relative to CONT, with greatest decreases for UFAA. Apparent total tract digestibilities of nutrient fractions were decreased by UFAA compared with TGA and SFAA and tended to be lower at the higher infusion amount. Apparent digestibility of total fatty acids (FA) was greater for SFAR than for SFAA. Plasma glucagon-like peptide-1 was greater for cows infused with UFAA than SFAA or TGA and increased at the higher amount. Plasma cholecystokinin was greater for cows infused with LCFA compared with CONT. Postruminal unsaturated FFA reduced intake and digestibility of nutrients and FA compared with postruminal TG infusion; saturated FA did not decrease dry matter intake or disrupt nutrient digestion. Glucagon-like peptide-1 may be involved in regulation of feed intake by long-chain fatty acids.


Dissecting structure-function of 3-O-sulfated heparin and engineered heparan sulfates.
Heparan sulfate (HS) polysaccharides are master regulators of diverse biological processes via sulfated motifs that can recruit specific proteins. 3-O-sulfation of HS/heparin is crucial for anticoagulant activity, but despite emerging evidence for roles in many other functions, a lack of tools for deciphering structure-function relationships has hampered advances. Here, we describe an approach integrating synthesis of 3-O-sulfated standards, comprehensive HS disaccharide profiling, and cell engineering to address this deficiency. Its application revealed previously unseen differences in 3-O-sulfated profiles of clinical heparins and 3-O-sulfotransferase (HS3ST)–specific variations in cell surface HS profiles. The latter correlated with functional differences in anticoagulant activity and binding to platelet factor 4 (PF4), which underlies heparin-induced thrombocytopenia, a known side effect of heparin. Unexpectedly, cells expressing the HS3ST4 isoenzyme generated HS with potent anticoagulant activity but weak PF4 binding. The data provide new insights into 3-O-sulfate structure-function and demonstrate proof of concept for tailored cell-based synthesis of next-generation heparins.


Expression and function of Pdcd4 in mouse endometrium during early pregnancy.
Embryo implantation is a complex process involving synchronised crosstalk between a receptive endometrium and functional blastocysts. Apoptosis plays an important role in this process as well as in the maintenance of pregnancy. In this study, we analysed the expression pattern of programmed cell death 4 (Pdcd4), a gene associated with apoptosis in the mouse endometrium, during early pregnancy and pseudopregnancy by real-time quantitative polymerase chain reaction, in situ hybridisation, Western blotting and immunohistochemistry. The results showed that Pdcd4 was increased along with days of pregnancy and significantly reduced at implantation sites (IS) from day 5 of pregnancy (D5). The level of Pdcd4 at IS was substantially lower than that at interimplantation sites (IIS) on D6 and D7. In addition, Pdcd4 expression in the endometrium was reduced in response to artificially induced decidualisation in vivo and in vitro Downregulation of Pdcd4 gene expression in cultured primary stromal cells promoted decidualisation, while upregulation inhibited the decidualisation process by increasing apoptosis. These results demonstrate that Pdcd4 is involved in stromal cell decidualisation by mediating apoptosis and therefore plays a role in embryo implantation in mice.


Inflammatory cytokine response and cardiac troponin I changes in cardiopulmonary bypass using two cardioplegia solutions; del Nido and modified St. Thomas': a randomized controlled trial.
Long intervals of del Nido (DN) solution administration, 90 minutes, may result in rewarming of the myocardial tissue and elevate metabolic demand and hypoxia. This will potentially increase inflammatory response due to ischemia-reperfusion injury. We conducted this study to compare the inflammatory response between patients receiving DN and multi-dose St Thomas' cardioplegia solution (MST) in cardiopulmonary bypass (CPB) surgery for the correction of tetralogy of Fallot (TF). Fifty-nine pediatric patients undergoing TF total correction surgery were randomly assigned into two groups: DN and MST. The patients' demographic data, blood chemistry parameters, hemodynamics and other clinical variables were recorded. TNF-a, IL-6, IL-8, IL-10 and cTnI were measured after anesthesia induction (before skin incision), immediately after cross-clamp removal and 24 hours after admission to the intensive care unit (ICU). Thirty-two patients of a mean age of 28.0±16.4 months received DN and 27 patients of a mean age of 24.2±15.9 months received MST. Perioperative clinical parameters were not significantly different between the two groups. Cytokine levels for all patients were significantly increased after surgery. Inter-group comparisons of cytokine levels demonstrated no significant differences in TNF-α, IL-6 and IL-8 cytokines levels. IL-10 level showed a moderately significant increase in the MST group compared to the DN group after surgery (2.94±0.9 vs. 2.46±0.61 log10 pg/mL, respectively; p=0.039). Postoperative lactate level was significantly different between two groups (2.475±1.29 vs 1.63±0.82 mg/dL in DN and MST groups, respectively; p=0.007). CTnI levels increased after surgery and remained constant until 24 hours after surgery. Significant differences between the MST and DN groups, at all times, were not detected. The anti-inflammatory cytokine response in the MST group is significantly better than in the DN group. This may be due to shorter intervals of the MST cardioplegia solution administration, which prevents rewarming of the myocardium, increased metabolic demand and hypoxia. Decreasing the intervals of DN administration may improve its cardioprotective properties.


Crowdsourcing Samples in Cognitive Science.
Crowdsourcing data collection from research participants recruited from online labor markets is now common in cognitive science. We review who is in the crowd and who can be reached by the average laboratory. We discuss reproducibility and review some recent methodological innovations for online experiments. We consider the design of research studies and arising ethical issues. We review how to code experiments for the web, what is known about video and audio presentation, and the measurement of reaction times. We close with comments about the high levels of experience of many participants and an emerging tragedy of the commons.


Extra electronic outer-shell peculiarities accessible under a joint XPS and DFT study.
Electronic configuration of chemically bound atoms at the surface, including adsorbed species, or in the bulk of a solid contains a set of natural traps for energy absorption provided by valence band transitions or plasmon oscillations. The core level excitation of any origin is generally coupled with those traps, forming a multichannel route for nonradiative energy dissipation. Using an example of Pt and graphite-based materials, the study shows experimental tracing over these channels by means of elastic electron scattering and X-ray photoelectron spectroscopy. As a complement to the experimental data, calculations of the density of states provide information on chemical behavior and local geometry of the atoms in a sample.


Higher prevalence of secretory CSE1L/CAS in sera of patients with metastatic cancer.
Metastatic markers are highly useful diagnostic and prognostic indicators of cancer metastasis. Herein, we report that secretory CSE1L/CAS, a cellular apoptosis susceptibility protein, is a new marker for metastatic cancer. CAS was colocalized with matrix metalloproteinase-2 in vesicles surrounding the outside of MCF-7 cell membranes, and the COOH-terminal domain of CAS was associated with matrix metalloproteinase-2-containing vesicles. Immunohistochemical staining for CAS was positive in the stroma and gland lumens of human metastatic cancer tissues. CAS was also detected in conditioned medium from B16-F10 melanoma cells and more frequently in the sera of patients with metastatic cancer than in sera from patients with primary cancer. Specifically, the prevalence of serum CAS in serum samples from 146 patients was 58.2% (32 of 55), 32.0% (8 of 25), and 12.1% (8 of 66) for patients with metastatic, invasive, and primary cancers, respectively. Our results suggest that CAS is a secretory protein associated with cancer metastasis, which may have clinical utility in metastatic cancer screening and diagnosis.


Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor.
Plasmin(ogen) receptors are expressed by many gram-positive and gram-negative bacteria. We previously isolated a plasmin receptor from a pathogenic group A streptococcal strain (C. C. Broder, R. Lottenberg, G. O. von Mering, K. H. Johnston, and M. D. P. Boyle, J. Biol. Chem. 266:4922-4928, 1991). The gene encoding this plasmin receptor, plr, was isolated from a lambda gt11 library of chromosomal DNA from group A streptococcal strain 64/14 by screening plaques with antibodies raised against the purified streptococcal plasmin receptor protein. The gene was subcloned by using a low-copy-number plasmid and stably expressed in Escherichia coli, resulting in the production of an immunoreactive and functional receptor protein. The DNA sequence of the gene contained an open reading frame encoding 335 amino acids with a predicted molecular weight of 35,787. Upstream of the open reading frame, putative promoter and ribosomal binding site sequences were identified. The experimentally derived amino acid sequences of the N terminus and three cyanogen bromide fragments of the purified streptococcal plasmin receptor protein corresponded to the predicted sequence encoded by plr. The deduced amino acid sequence for the plasmin receptor protein revealed significant similarity (39 to 54% identical amino acid residues) to glyceraldehyde 3-phosphate dehydrogenases.


A Novel Physical Activity and Sedentary Behavior Classification and Its Relationship With Physical Fitness in Spanish Older Adults: The PHYSMED Study.
As there is no gold-standard methodology to classify older people in relation to physical activity (PA) and sedentary behavior (SB), this paper aimed to propose a classification combining PA and SB. Within a broader study, 433 subjects, aged 55 years and older (57% females) from Madrid and Majorca, were evaluated for PA and SB by means of validated questionnaires. Physical fitness was analyzed objectively using the EXERNET test battery. Cluster analyses were used to establish behavioral patterns, combining PA and SB. Males spent more time doing regular PA but less time walking and working at home than females (P < .001). Comparing the groups (inactive and high sedentary, inactive and low sedentary, active and high sedentary, and active and low sedentary), the worst aerobic endurance (P < .001) and lower body strength (P < .05) were obtained in males from both inactive groups. Agility was highest in the active and low sedentary group (P < .05). No significant differences were observed in females. The proposed classification is valid, as it classifies subjects according to their PA and SB, and outcomes are related to objectively measured fitness. It could facilitate the work of public health authorities, researchers, and physicians.


Three-dimensional electron realm in VSe2 by soft-x-ray photoelectron spectroscopy: origin of charge-density waves.
The resolution of angle-resolved photoelectron spectroscopy (ARPES) in three-dimensional (3D) momentum k is fundamentally limited by ill defined surface-perpendicular wave vector k(perpendicular) associated with the finite photoelectron mean free path. Pushing ARPES into the soft-x-ray energy region sharpens the k(perpendicular) definition, allowing accurate electronic structure investigations in 3D materials. We apply soft-x-ray ARPES to explore the 3D electron realm in a paradigm transition metal dichalcogenide VSe2. Essential to break through the dramatic loss of the valence band photoexcitation cross section at soft-x-ray energies is the advanced photon flux performance of our synchrotron instrumentation. By virtue of the sharp 3D momentum definition, the soft-x-ray ARPES experimental band structure and Fermi surface of VSe2 show a textbook clarity. We identify pronounced 3D warping of the Fermi surface and show that its concomitant nesting acts as the precursor for the exotic 3D charge-density waves in VSe2. Our results demonstrate the immense potential of soft-x-ray ARPES to explore details of 3D electronic structure.


Genomic data illuminates demography, genetic structure and selection of a popular dog breed.
Genomic methods have proved to be important tools in the analysis of genetic diversity across the range of species and can be used to reveal processes underlying both short- and long-term evolutionary change. This study applied genomic methods to investigate population structure and inbreeding in a common UK dog breed, the Labrador Retriever. We found substantial within-breed genetic differentiation, which was associated with the role of the dog (i.e. working, pet, show) and also with coat colour (i.e. black, yellow, brown). There was little evidence of geographical differentiation. Highly differentiated genomic regions contained genes and markers associated with skull shape, suggesting that at least some of the differentiation is related to human-imposed selection on this trait. We also found that the total length of homozygous segments (runs of homozygosity, ROHs) was highly correlated with inbreeding coefficient. This study demonstrates that high-density genomic data can be used to quantify genetic diversity and to decipher demographic and selection processes. Analysis of genetically differentiated regions in the UK Labrador Retriever population suggests the possibility of human-imposed selection on craniofacial characteristics. The high correlation between estimates of inbreeding from genomic and pedigree data for this breed demonstrates that genomic approaches can be used to quantify inbreeding levels in dogs, which will be particularly useful where pedigree information is missing.


Private algorithms for the protected in social network search.
Motivated by tensions between data privacy for individual citizens and societal priorities such as counterterrorism and the containment of infectious disease, we introduce a computational model that distinguishes between parties for whom privacy is explicitly protected, and those for whom it is not (the targeted subpopulation). The goal is the development of algorithms that can effectively identify and take action upon members of the targeted subpopulation in a way that minimally compromises the privacy of the protected, while simultaneously limiting the expense of distinguishing members of the two groups via costly mechanisms such as surveillance, background checks, or medical testing. Within this framework, we provide provably privacy-preserving algorithms for targeted search in social networks. These algorithms are natural variants of common graph search methods, and ensure privacy for the protected by the careful injection of noise in the prioritization of potential targets. We validate the utility of our algorithms with extensive computational experiments on two large-scale social network datasets.


Regional cerebral metabolism in female alcoholics of moderate severity does not differ from that of controls.
It is generally believed that women are more vulnerable to alcohol's toxic effects than men. Studies in male alcoholics have consistently shown reductions in brain glucose metabolism. However, such studies have not been done in female alcoholics. The purpose of this study was to evaluate if similar or worse brain metabolic abnormalities occurred in female alcoholics. For this purpose, we measured regional brain metabolism with positron emission tomography and [18F]fluorodeoxyglucose in 10 recently detoxified female alcoholics and compared it with that in 12 age-matched female controls. There were no differences between alcoholics and control females in regional brain glucose metabolism whether we used regions of interest analysis or statistical parameter maps methods. These results do not support a higher toxicity for the effects of alcohol in the female brain, as assessed with regional brain glucose metabolism, because metabolic values in female alcoholics did not differ from those of controls, whereas metabolic values in male alcoholics are generally lower than those in controls. However, this study is confounded by the fact that the severity of alcohol use in these female alcoholics was less than that of the male alcoholics previously investigated in positron emission tomography studies. Future studies in male subjects with alcoholism of moderate severity are required to address gender differences in sensitivity to alcohol effects in brain metabolism.


Weather-Based Forecasting of Rhizoctonia Web Blight Development on Container-Grown Azalea.
The most reliable approach for timing fungicides to control Rhizoctonia web blight on container-grown azalea has been a fixed calendar date. The purpose of this study was to model periods when a preventive control could be applied in advance of significant increases in leaf blight intensity (LBI) in response to a daily accumulated risk value indicating conducive conditions of temperature (18 or more hours between 20 and 30°C when maximum temperature is less than 35°C), leaf wetness (16 or more hours), and/or rainfall (greater than 6.7 mm above the maximum daily irrigation). Analysis used weather measurements taken every 30 minutes from 11 site-year datasets from 2006 to 2011, and weekly or biweekly LBI assessments. Six developmental site-year datasets were analyzed using receiver operating characteristic (ROC) curve procedures to evaluate models. Only a single model predicted true positives and negatives significantly greater than a 50:50 probability. Subsequently, five site-year validation datasets were evaluated. Similar results were obtained from both the developmental and validation datasets, which produced ROC areas of 0.7715 (P = 0.0120) and 0.8393 (P = 0.0034), respectively. The practical application of weather-based forecasting for management of web blight in nurseries is discussed.


Hrs and SNX3 functions in sorting and membrane invagination within multivesicular bodies.
After internalization, ubiquitinated signaling receptors are delivered to early endosomes. There, they are sorted and incorporated into the intralumenal invaginations of nascent multivesicular bodies, which function as transport intermediates to late endosomes. Receptor sorting is achieved by Hrs--an adaptor--like protein that binds membrane PtdIns3P via a FYVE motif-and then by ESCRT complexes, which presumably also mediate the invagination process. Eventually, intralumenal vesicles are delivered to lysosomes, leading to the notion that EGF receptor sorting into multivesicular bodies mediates lysosomal targeting. Here, we report that Hrs is essential for lysosomal targeting but dispensable for multivesicular body biogenesis and transport to late endosomes. By contrast, we find that the PtdIns3P-binding protein SNX3 is required for multivesicular body formation, but not for EGF receptor degradation. PtdIns3P thus controls the complementary functions of Hrs and SNX3 in sorting and multivesicular body biogenesis.


Neuroendocrine changes in the aging reproductive axis of female rhesus macaques (Macaca mulatta).
Femalerhesus macaques show monthly menstrual cycles and eventually enter menopause at approximately 25 yr of age. To help identify early biomarkers of menopause in this nonhuman primate, we monitored reproductive hormones longitudinally from aged female macaques during the transitions from premenopause to perimenopause and postmenopause and found that, indeed, elevated plasma FSH was a better predictive factor of menopause onset than age. In a second experiment, we compared reproductive hormone profiles of young adult macaques (8-10 yr old) with those of regularly cycling old macaques (approximately 24 yr old). Indwelling vascular catheters were used for remote blood collection for at least 100 consecutive days, thereby covering three complete menstrual cycles in each macaque. Plasma levels of estradiol, progesterone, LH, FSH, follicular phase inhibin B, and anti-müllerian hormone (AMH) were determined during each menstrual cycle and were averaged for each animal; group mean differences were analyzed using one-way ANOVA. Old premenopausal macaques showed regular menstrual cycles that were qualitatively indistinguishable from those of young macaques; peak plasma levels of estradiol, progesterone, and LH were not significantly different. In marked contrast, peak plasma FSH concentrations were significantly higher, while inhibin B and AMH levels were generally lower, in the old premenopausal macaques compared with those in the young macaques. These data provide further evidence that rhesus macaques serve as an excellent model to study underlying mechanisms of human menopause. Furthermore, the data suggest that an age-related change in FSH, inhibin B, and AMH secretion may be the first endocrine manifestation of the transition into perimenopause, potentially having value in predicting the onset of the perimenopausal transition.