Sociocultural influences on mental health service use by Latino older adults for emotional distress: exploring the mediating and moderating role of informal social support.
This study investigated the determinants of mental health services for emotional distress by low-income older Latinos living in Los Angeles County (United States). The functional effects of informal support on service use were tested while accounting for dimensions of support and sociocultural factors. Using data from a cross-sectional survey of older Latinos (n = 235), we preformed a secondary data analysis using path analysis with Poisson regression to assess mediation and moderation models by type of perceived support, emotional and instrumental. Data was originally collected between 1998 and 2005. Results indicated that no mediation effects were present however significant moderation effects emerged. Respondents with low levels of linguistic acculturation and informal support used fewer services. Variations by type of informal support emerged, with emotional support presenting the best model fit compared to instrumental support in the moderation model analysis. The direct effects of linguistic acculturation and age remained significant after controlling for need. Findings reveal limited informal support is a risk factor for the underutilization of mental health services by older Latinos with low levels of linguistic acculturation as they age. Integrating support resources that assist in navigating service systems and address language barriers can enhance service use by older Latino adults. Implications for service delivery and program development are discussed.


Effect of load carriage on chronic low back pain in adults with cerebral palsy.
Chronic low back pain (LBP) is a common secondary musculoskeletal problem among adults with cerebral palsy (CP). This study investigated the feasibility of incorporating backpack carriage with daily production activities for relieving chronic LBP among adults with CP. The effects of backpack carriage in relieving chronic LBP for adults with CP. A multiple bivariate approach with convenience sampling. Nine adults with CP suffering from chronic LBP were scheduled to carry a 4 lb backpack at work for 60 minutes on weekdays for four consecutive weeks. The pain level was rated by the participants on an 11-point scale. Muscle activity of erector spinae was measured by surface electromyography. Significant improvement in back pain was found immediately after the backpack carriage with no adverse effect reported. It was accompanied with significantly reduced erector spinae activity. Although the overall change in pain ratings across the study period was not significant, a decreasing trend with time was observed. The loaded backpack has the potential to serve as an 'orthosis' for immediate back pain relief among adults with CP. Its underlying mechanism can be attributed to a reduction in back muscle tension during the load carriage.


Moderate resistance training and vascular health in overweight women.
Impaired endothelial function has been implicated as an initial step in the pathogenesis of atherosclerosis leading to cardiovascular disease (CVD). Aerobic exercise improves vascular function and reduces risk of CVD morbidity and mortality; however, the effects of resistance training on vascular function remains unclear. Thus, the aim of this study was to examine the effects of 1 yr of resistance training (RT) on vascular structure and function in overweight but otherwise healthy eumenorrheic women. Study participants consisted of 30 (15 control, 15 RT) overweight (BMI > 25 kg x m(-2)) women, aged 24-44 yr, studied before and after a 1-yr RT intervention. Vascular structure and function were assessed via noninvasive ultrasound imaging of the carotid and brachial arteries, respectively. Body composition, blood pressure, and fasting blood lipids and glucose also were measured. The RT group demonstrated a significant mean improvement in one-repetition maximum bench press following 1 yr of RT (P < 0.05). There also was a significant increase in lean body mass in the RT group compared with the control group (P = 0.04). There were no training-associated changes in blood pressure, fasting blood lipids, glucose, or insulin levels. Although there was no change in carotid artery intimamedia thickness, peak flow-mediated dilation significantly improved in the RT group (P < 0.05). The principal significant finding in this study was the initial demonstration that RT alone can improve brachial artery endothelial function in overweight eumenorrheic women.


Systematic review of physical activity and health in the early years (aged 0-4 years).
The early years represent a critical period for promoting physical activity. However, the amount of physical activity needed for healthy growth and development is not clear. Using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) framework, we aimed to present the best available evidence to determine the relationship between physical activity and measures of adiposity, bone and skeletal health, motor skill development, psychosocial health, cognitive development, and cardiometabolic health indicators in infants (1 month - 1 year), toddlers (1.1-3.0 years), and preschoolers (3.1-4.9 years). Online databases, personal libraries, and government documents were searched for relevant studies. Twenty-two articles, representing 18 unique studies and 12 742 enrolled participants, met inclusion criteria. The health indicators of interest were adiposity (n = 11), bone and skeletal health (n = 2), motor development (n = 4), psychosocial health (n = 3), cognitive development (n = 1), and cardiometabolic health indicators (n = 3); these indicators were pre-specified by an expert panel. Five unique studies involved infants, 2 involved toddlers, and 11 involved preschoolers. In infants, there was low- to moderate-quality evidence to suggest that increased or higher physical activity was positively associated with improved measures of adiposity, motor skill development, and cognitive development. In toddlers, there was moderate-quality evidence to suggest that increased or higher physical activity was positively associated with bone and skeletal health. In preschoolers, there was low- to high-quality evidence on the relationship between increased or higher physical activity and improved measures of adiposity, motor skill development, psychosocial health, and cardiometabolic health indicators. There was no serious inconsistency in any of the studies reviewed. This evidence can help to inform public health guidelines. (PROSPERO registration: CRD42011001243).


Electrophysiological abnormalities in the transplanted human heart.
Fourteen relatively long term survivors of cardiac transplantation underwent systematic electrophysiological evaluation and ambulatory electrocardiographic monitoring. Six patients had prolonged conduction intervals during sinus rhythm. Sinus node function could be assessed in all donor atria and in 10 recipient atria. Sinus node recovery times were prolonged in four of the donor atria and in six recipient atria. In the donor atria abnormalities of sinus node automaticity were invariably associated with abnormalities of sinoatrial conduction. Four patients showed functional duality of atrioventricular nodal conduction during programmed extrastimulation, but no patient developed re-entrant arrhythmia. During ambulatory electrocardiographic monitoring no pronounced tachyarrhythmias were recorded. Three patients showed abnormalities of sinus node impulse formation. All three patients had abnormal sinus node recovery times during their electrophysiological study. Long term survivors of cardiac transplantation have a high incidence of electrophysiological abnormalities. Abnormalities of donor sinus node function are probably of clinical significance. The clinical significance of abnormalities detected within the atrioventricular conduction system of the denervated heart remains to be elucidated.


Comparisons of the Factor Structure and Measurement Invariance of the Spence Children's Anxiety Scale-Parent Version in Children with Autism Spectrum Disorder and Typically Developing Anxious Children.
The Spence Children's Anxiety Scale-Parent version (SCAS-P) is often used to assess anxiety in children with autism spectrum disorder (ASD), however, little is known about the validity of the tool in this population. The aim of this study was to determine whether the SCAS-P has the same factorial validity in a sample of young people with ASD (n = 285), compared to a sample of typically developing young people with anxiety disorders (n = 224). Poor model fit with all of the six hypothesised models precluded invariance testing. Exploratory factor analysis indicated that different anxiety phenomenology characterises the two samples. The findings suggest that cross-group comparisons between ASD and anxious samples based on the SCAS-P scores may not always be appropriate.


Open pneumothorax: the spectrum and outcome of management based on Advanced Trauma Life Support recommendations.
The current management of open pneumothorax (OPTX) is based on Advanced Trauma Life Support (ATLS) recommendations and consists of the application of a three-way occlusive dressing, followed by intercostal chest drain insertion. Very little is known regarding the spectrum and outcome of this approach, especially in the civilian setting. We conducted a retrospective review of 58 consecutive patients with OPTX over a four-year period managed in a high volume metropolitan trauma service in South Africa. Of the 58 patients included, 95% (55/58) were male, and the mean age for all patients was 21 years. Ninety-seven percent of all injuries were inflicted by knives and the remaining 3% (2/58) of injuries were inflicted by unknown weapons. 59% of injuries were left sided. In six patients (10%) a protocol violation was present in their management. Five of the six patients (83%) in whom protocol violation occurred developed a life-threatening event (tension PTX) compared to none amongst those where the protocol was followed (p < 0.001). There was no mortality as a direct result of management of OPTX following ATLS recommendations. ATLS recommendations for OPTX are safe and effective. Any deviation from this standard practice is associated with avoidable morbidity and potential mortality.


Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia II: developing imaging biomarkers to enhance treatment development for schizophrenia and related disorders.
The Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia (CNTRICS) initiative, funded by an R13 from the National Institute of Mental Health, seeks to enhance translational research in treatment development for impaired cognition in schizophrenia by developing tools from cognitive neuroscience into useful measures of treatment effects on behavior and brain function. An initial series of meetings focused on the selection of a new set of tasks from cognitive neuroscience for the measurement of treatment effects on specific cognitive and neural systems. Subsequent validation and optimization studies are underway and a subset of validated measures with well-characterized psychometric properties will be generally available in 2011. This article describes results of the first meeting of the second phase of the Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia, which seeks to develop imaging biomarkers and improved animal models to enhance translational research. In this meeting, we considered issues related to the use of methods such as functional magnetic resonance imaging, electroencephalography, magnetoencephalography, and transcranial magnetic simulation as biomarkers for treatment development. We explored the biological nature of the signals measured by each method, their validity and reliability as measures of cognition-related neural activity, potential confounds related to drug effects on the signal of interest, and conceptual, methodological, and pragmatic issues related to their use in preclinical, first into human, and multicenter phase II and III studies. This overview article describes the background and goals of the meeting together with a summary of the major issues discussed in more detail in the accompanying articles appearing in this issue of Biological Psychiatry.


Diffuse hypoplasia of the aortic arch and isthmus in a patient with Williams syndrome.
Williams syndrome is a rare neurodevelopmental disorder characterized by mental retardation, growth deficiency, hypercalcemia, cardiac defects, and a distinctive facial appearance. Cardiovascular abnormalities are present in approximately 80% of Williams syndrome patients. Surgical treatment is generally performed for supravalvular aortic stenosis, aortic coarctation, pulmonary artery stenosis, or ventricular septal defect. In rare cases, diffuse hypoplasia of the aortic arch with a normal left ventricular outflow tract and ascending aorta may be diagnosed in early childhood. Described herein is the case of a 16-month-old female with Williams syndrome and diffuse hypoplasia of the aortic arch and isthmus, and concomitant pulmonary stenosis and a ventricular septal defect. The patient underwent a successful surgical repair of the aortic arch with a modified pericardial patch technique.


Defining seasonal marine microbial community dynamics.
Here we describe, the longest microbial time-series analyzed to date using high-resolution 16S rRNA tag pyrosequencing of samples taken monthly over 6 years at a temperate marine coastal site off Plymouth, UK. Data treatment effected the estimation of community richness over a 6-year period, whereby 8794 operational taxonomic units (OTUs) were identified using single-linkage preclustering and 21 130 OTUs were identified by denoising the data. The Alphaproteobacteria were the most abundant Class, and the most frequently recorded OTUs were members of the Rickettsiales (SAR 11) and Rhodobacteriales. This near-surface ocean bacterial community showed strong repeatable seasonal patterns, which were defined by winter peaks in diversity across all years. Environmental variables explained far more variation in seasonally predictable bacteria than did data on protists or metazoan biomass. Change in day length alone explains >65% of the variance in community diversity. The results suggested that seasonal changes in environmental variables are more important than trophic interactions. Interestingly, microbial association network analysis showed that correlations in abundance were stronger within bacterial taxa rather than between bacteria and eukaryotes, or between bacteria and environmental variables.


Iron-sulfur cluster biosynthesis: toward an understanding of cellular machinery and molecular mechanism.
Iron-sulfur clusters are among the most complex metal-containing prosthetic centers in biology. Most if not all of the proteins involved in the biosynthesis of "simple" Fe-S clusters have been identified. The structural and functional chemistry of these proteins has been the subject of intense research efforts, and many of the key details are now understood in structural and mechanistic detail. The fact that Fe-S cluster-binding proteins can be reconstituted in vitro with no accessory proteins provides an important indicator of the intracellular roles for many proteins on the Fe-S cluster assembly pathway. Indeed, such proteins are more correctly viewed as carrier proteins, rather than as catalysts for the reaction, that both avoid the toxicity associated with free iron and sulfide and allow delivery at lower intracellular concentrations of these species. The IscU (or ISU) family of proteins serves a key role as scaffolding proteins on which [2Fe-2S] building blocks are assembled prior to transfer to final apo target proteins. IscU in particular exhibits highly unusual conformational flexibility that appears critical to its function.


3.3 Å structure of Niemann-Pick C1 protein reveals insights into the function of the C-terminal luminal domain in cholesterol transport.
Niemann-Pick C1 (NPC1) and NPC2 proteins are indispensable for the export of LDL-derived cholesterol from late endosomes. Mutations in these proteins result in Niemann-Pick type C disease, a lysosomal storage disease. Despite recent reports of the NPC1 structure depicting its overall architecture, the function of its C-terminal luminal domain (CTD) remains poorly understood even though 45% of NPC disease-causing mutations are in this domain. Here, we report a crystal structure at 3.3 Å resolution of NPC1* (residues 314-1,278), which-in contrast to previous lower resolution structures-features the entire CTD well resolved. Notably, all eight cysteines of the CTD form four disulfide bonds, one of which (C909-C914) enforces a specific loop that in turn mediates an interaction with a loop of the N-terminal domain (NTD). Importantly, this loop and its interaction with the NTD were not observed in any previous structures due to the lower resolution. Our mutagenesis experiments highlight the physiological relevance of the CTD-NTD interaction, which might function to keep the NTD in the proper orientation for receiving cholesterol from NPC2. Additionally, this structure allows us to more precisely map all of the disease-causing mutations, allowing future molecular insights into the pathogenesis of NPC disease.


Assessment of risk area during coronary occlusion and infarct size after reperfusion with myocardial contrast echocardiography using left and right atrial injections of contrast.
Myocardial opacification during echocardiography has been demonstrated after left (LA) and right (RA) atrial injections of contrast, and microvascular damage with reduced blood flow and impaired flow reserve has been documented in necrotic myocardial tissue. Therefore, we hypothesized that because of its ability to depict capillary perfusion, myocardial contrast echocardiography (MCE) can be used to define risk area during coronary occlusion and infarct size after reperfusion with LA and RA injections of contrast in the presence of pharmacologically induced coronary hyperemia. Eighteen open-chest anesthetized dogs with 3 to 6 hours of left anterior descending artery occlusion and 15 minutes of reflow were studied in the presence of either dipyridamole (0.56 mg/kg over a period of 4 minutes) or dobutamine (15 micrograms.kg-1.min-1). Technetium autoradiography was performed for risk area assessment; infarct size was measured with triphenyl tetrazolium chloride; and in 11 dogs, myocardial blood flow was measured with radiolabeled microspheres. A close linear relation was noted between the MCE defect size and autoradiographic risk area during coronary occlusion both during LA (y = 0.95x-0.25, r = .97, P < .001) and RA (y = 0.90x+0.98, r = .86, P < .001) injections of contrast. During reperfusion, the contrast defect size on MCE was always less transmural than during occlusion and correlated closely with infarct size during both LA (y = 1.07x-2.37, r = .98, P < .001) and RA (y = 1.02x-0.61, r = .95, P < .001) injections of contrast. In the 11 dogs in whom radiolabeled microsphere-derived blood flow was measured during reperfusion, an inverse relation was noted between infarct size and transmural blood flow (y = -1.12x+121, r = -.95, P = .001), implying that MCE defects after reperfusion indicate necrotic regions with reduced blood flow or impaired microvascular flow reserve. A close linear relation (y = 0.79x-0.001, r = .98, P < .001) was also noted between endocardial/epicardial ratio of background-subtracted peak video intensity on MCE and endocardial/epicardial blood flow ratio in the eight dogs with infarction who underwent this measurement after reperfusion. MCE performed with LA and RA injections of contrast in the presence of pharmacologically induced coronary hyperemia can be used to determine, in vivo, the risk area during coronary occlusion and infarct size after reperfusion. These results could have important implications in this era of myocardial reperfusion.


GFAP isoforms control intermediate filament network dynamics, cell morphology, and focal adhesions.
Glial fibrillary acidic protein (GFAP) is the characteristic intermediate filament (IF) protein in astrocytes. Expression of its main isoforms, GFAPα and GFAPδ, varies in astrocytes and astrocytoma implying a potential regulatory role in astrocyte physiology and pathology. An IF-network is a dynamic structure and has been functionally linked to cell motility, proliferation, and morphology. There is a constant exchange of IF-proteins with the network. To study differences in the dynamic properties of GFAPα and GFAPδ, we performed fluorescence recovery after photobleaching experiments on astrocytoma cells with fluorescently tagged GFAPs. Here, we show for the first time that the exchange of GFP-GFAPδ was significantly slower than the exchange of GFP-GFAPα with the IF-network. Furthermore, a collapsed IF-network, induced by GFAPδ expression, led to a further decrease in fluorescence recovery of both GFP-GFAPα and GFP-GFAPδ. This altered IF-network also changed cell morphology and the focal adhesion size, but did not alter cell migration or proliferation. Our study provides further insight into the modulation of the dynamic properties and functional consequences of the IF-network composition.


Effects of chronic inhibition of GABA synthesis on attention and impulse control.
Cortical GABA regulates a number of cognitive functions including attention and working memory and is dysregulated in a number of psychiatric conditions. In schizophrenia for example, changes in GABA neurons [reduced expression of glutamic acid decarboxylase (GAD), parvalbumin (PV) and the GABA reuptake transporter (GAT1)] suggest reduced cortical GABA synthesis and release; these changes are hypothesized to cause the cognitive deficits observed in this disorder. The goals of this experiment were to determine whether chronically reducing GAD function within the rat PFC causes attention deficits and alterations in PV and GAT1 expression. Male Sprague Dawley rats were trained on the 5-choice serial reaction time task (5CSRTT, a task of attention) until they reached criterion performance and then were implanted with a bilateral cannula aimed at the medial PFC. Cannulae were connected to osmotic minipumps that infused the GAD inhibitor l-allylglycine (LAG, 3.2μg/0.5μl/h) for 13days. Following a 5-day recovery from surgery rats were tested on the standard 5CSRTT for 5 consecutive days and then tested on two modifications of the 5CSRTT. Finally, locomotor activity was assessed and the rats sacrificed. Brains were rapidly extracted and flash frozen and analyzed for the expression of GAD67, PV, GAT1 and the obligatory NMDA receptor subunit NR1. Chronic LAG infusions transiently impaired attention, persistently impaired impulse control and increased locomotor activity. Behavioral changes were associated with an upregulation of GAD67, but no change in PV, GAT1 or NR1 expression. Chronic inhibition of GABA synthesis within the medial PFC, increased impulsive behavior and locomotion, but did not impair attention; results consistent with previous research following acute inhibition of GABA synthesis. Moreover, our data do not support the hypothesis that decreasing GABA synthesis and release is sufficient to cause changes in other GABA-related proteins.


Crystal structure of human XLF: a twist in nonhomologous DNA end-joining.
DNA double-strand breaks represent one of the most severe forms of DNA damage in mammalian cells. One pathway for repairing these breaks occurs via nonhomologous end-joining (NHEJ) and depends on XRCC4, LigaseIV, and Cernunnos, also called XLF. Although XLF stimulates XRCC4/LigaseIV to ligate mismatched and noncohesive DNA ends, the mechanistic basis for this function remains unclear. Here we report the structure of a partially functional 224 residue N-terminal fragment of human XLF. Despite only weak sequence similarity, XLF(1-170) shares structural homology with XRCC4(1-159). However, unlike the highly extended 130 A helical domain observed in XRCC4, XLF adopts a more compact, folded helical C-terminal region involving two turns and a twist, wrapping back to the structurally conserved N terminus. Mutational analysis of XLF and XRCC4 reveals a potential interaction interface, suggesting a mechanism for how XLF stimulates the ligation of mismatched ends.


Autoimmune basis for postural tachycardia syndrome.
Patients with postural tachycardia syndrome (POTS) have exaggerated orthostatic tachycardia often following a viral illness, suggesting autoimmunity may play a pathophysiological role in POTS. We tested the hypothesis that they harbor functional autoantibodies to adrenergic receptors (AR). Fourteen POTS patients (7 each from 2 institutions) and 10 healthy subjects were examined for α1AR autoantibody-mediated contractility using a perfused rat cremaster arteriole assay. A receptor-transfected cell-based assay was used to detect the presence of β1AR and β2AR autoantibodies. Data were normalized and expressed as a percentage of baseline. The sera of all 14 POTS patients demonstrated significant arteriolar contractile activity (69±3% compared to 91±1% of baseline for healthy controls, P<0.001) when coexisting β2AR dilative activity was blocked; and this was suppressed by α1AR blockade with prazosin. POTS sera acted as a partial α1AR antagonist significantly shifting phenylephrine contractility curves to the right. All POTS sera increased β1AR activation (130±3% of baseline, P<0.01) and a subset had increased β2AR activity versus healthy subjects. POTS sera shifted isoproterenol cAMP response curves to the left, consistent with enhanced β1AR and β2AR agonist activity. Autoantibody-positive POTS sera demonstrated specific binding to β1AR, β2AR, and α1AR in transfected cells. POTS patients have elevated α1AR autoantibodies exerting a partial peripheral antagonist effect resulting in a compensatory sympathoneural activation of α1AR for vasoconstriction and concurrent βAR-mediated tachycardia. Coexisting β1AR and β2AR agonistic autoantibodies facilitate this tachycardia. These findings may explain the increased standing plasma norepinephrine and excessive tachycardia observed in many POTS patients.


Prediagnostic circulating carotenoid levels and the risk of non-Hodgkin lymphoma: the Multiethnic Cohort.
This analysis examined the association of non-Hodgkin lymphoma (NHL) with prediagnostic carotenoid levels, a marker for a diet rich in fruits and vegetables. We conducted a nested case-control study within the Multiethnic Cohort with 271 NHL cases and 538 controls matched on sex, ethnicity, location (Hawaii or Los Angeles), birth year, date and time of blood draw, and hours fasting before blood draw. Serum carotenoid levels were obtained by high-pressure liquid chromatography with photodiode array detection. Conditional logistic regression was used to calculate odds ratios (ORs) according to tertiles of serum carotenoids and trend tests using continuous variables. Higher total serum carotenoids (OR(T3 vs T1) = 0.66 [0.46-0.96]; P(trend) = .02), lycopene (OR = 0.54 [0.38-0.78]; P(trend) = .003), and α-cryptoxanthin (OR = 0.53 [0.36-0.78]; P(trend) = .003) were associated with a lower risk of NHL. For retinol (OR = 0.90 [0.61-1.33]; P(trend) = .04), a statistically significant inverse linear trend was detected. Risk estimates remained unchanged with adjustment for NHL risk factors and were similar in analyses stratified by sex and ethnicity; heterogeneity with NHL subtype was detected only for β-carotene. Other carotenoids, including α-carotene, β-carotene, lutein, β-cryptoxanthin, and zeaxanthin, showed no association with risk. These data provide support for a protective role of carotenoid-rich fruits and vegetables in the etiology of NHL.


Nivalenol-type populations of Fusarium graminearum and F. asiaticum are prevalent on wheat in southern Louisiana.
U.S. populations of the Fusarium graminearum clade cause head blight on wheat and barley and usually contaminate grain with the trichothecene mycotoxin deoxynivalenol (DON). Recently, however, individual nivalenol (NIV)-type isolates from the United States were described that belonged to either the newly described species F. gerlachii or the genetically distinct Gulf Coast population of F. graminearum sensu stricto (s.s.). Here, we describe the discovery of NIV-type F. graminearum s.s. populations that were found in high proportion (79%) among isolates from small-grain-growing regions of Louisiana. We genotyped 237 isolates from Louisiana with newly developed polymerase chain reaction (PCR) restriction fragment length polymorphism markers and multiplex PCR primers that distinguish among the three trichothecene types: the two DON types (15ADON and 3ADON) and NIV. These isolates were compared with 297 isolates from 11 other U.S. states, predominantly from the Midwest. Using Bayesian-model-based clustering, we discovered a southern Louisiana population of F. graminearum s.s. that was genetically distinct from the previously recognized pathogen population in the Midwest (MW15ADON population). Population membership was correlated with trichothecene type. Most isolates from the southern Louisiana population were of the NIV type, while the majority of the isolates from the Midwest were of the 15ADON type. A smaller proportion of isolates from Louisiana belonged to the previously described Gulf Coast population that was mostly of the 3ADON type. The NIV type was also identified in collections from Arkansas (12%), North Carolina (40%), and Missouri (2%), with the collections from Arkansas and North Carolina being small and unrepresentative. F. asiaticum was detected from the two southern Louisiana parishes Acadia and Alexandria. All identified 41 F. asiaticum isolates were of the NIV type. Greenhouse tests indicated that U.S. NIV types accumulated four times less trichothecene toxin than DON types on inoculated wheat. This is the first report of NIV-type populations of F. graminearum s. s. and F. asiaticum in the United States.


Navigating cancer network attractors for tumor-specific therapy.
Cells employ highly dynamic signaling networks to drive biological decision processes. Perturbations to these signaling networks may attract cells to new malignant signaling and phenotypic states, termed cancer network attractors, that result in cancer development. As different cancer cells reach these malignant states by accumulating different molecular alterations, uncovering these mechanisms represents a grand challenge in cancer biology. Addressing this challenge will require new systems-based strategies that capture the intrinsic properties of cancer signaling networks and provide deeper understanding of the processes by which genetic lesions perturb these networks and lead to disease phenotypes. Network biology will help circumvent fundamental obstacles in cancer treatment, such as drug resistance and metastasis, empowering personalized and tumor-specific cancer therapies.


Reinforcing mechanisms of single-walled carbon nanotube-reinforced polymer composites.
The reinforcing mechanisms of single-walled carbon nanotube-reinforced epoxy composites were studied by micromechanics models. The modeling results obtained from both Halpin-Tsai and Mori-Tanaka models are in good agreement with the experimental results. It has been found that these two models are also applicable to other single-walled carbon nanotube-reinforced, amorphous-polymer composites, given the existence of efficient load transfer. The reinforcing mechanisms that work in polymer-carbon nanotube composites were studied. The reasons responsible for the low mechanical property enhancement of single-walled carbon nanotube in polymer composites were discussed in conjunction with the effective fiber length concept, interface between nanotube bundles and the matrix, properties of the reinforcements and matrix, bundle effects, bundle curvature, and alignment.


The big and the small: challenges of imaging the brain's circuits.
The relation between the structure of the nervous system and its function is more poorly understood than the relation between structure and function in any other organ system. We explore why bridging the structure-function divide is uniquely difficult in the brain. These difficulties also explain the thrust behind the enormous amount of innovation centered on microscopy in neuroscience. We highlight some recent progress and the challenges that remain.


Infrared- and Raman-spectroscopy measurements of a transition in the crystal structure and a closing of the energy gap of BiTeI under pressure.
BiTeI is a giant Rashba spin splitting system, in which a noncentrosymmetric topological phase has recently been suggested to appear under high pressure. We investigated the optical properties of this compound, reflectivity and transmission, under pressures up to 15 GPa. The gap feature in the optical conductivity vanishes above p∼9 GPa and does not reappear up to at least 15 GPa. The plasma edge, associated with intrinsically doped charge carriers, is smeared out through a phase transition at 9 GPa. Using high-pressure Raman spectroscopy, we follow the vibrational modes of BiTeI, providing additional clear evidence that the transition at 9 GPa involves a change of crystal structure. This change of crystal structure possibly inhibits the high-pressure topological phase from occurring.


Polymorphisms in arsenic metabolism genes, urinary arsenic methylation profile and cancer.
Arsenic-metabolism-related genes can regulate the arsenic methylation process and may influence susceptibility to cancer. We evaluated the roles of arsenic metabolism genes on urinary arsenic profiles of repeated measurement with 15-year follow-up (1988-2004) through general linear model (GLM) and assessed the effect of the changed extent of urinary arsenic profiles on cancer risk. Questionnaire information and blood samples and two urines (1988 and 2004) were collected from 208 subjects in an arseniasis hyperendemic area in Taiwan. Profiles for concentrations of urinary arsenic were determined using HPLC-HG-AAS. The relative proportion of each arsenic species was calculated by dividing the concentration of each arsenic species by the total arsenic concentration. Genotyping was done using the 5' nuclease allelic discrimination (Taqman) assay. The incidence of cancer was identified through linking to the National Cancer Registry Systems. The Cox proportional hazards model and survival curves were used in the analyses. After a 15-year follow-up, baseline monomethylarsonic acid percentage (MMA%) and change in MMA% exhibited a significant dose-response relationship with cancer risk. Individuals with a higher baseline MMA% and a lower change in MMA% had the earliest cancer incidence (statistically significant). Through GLM, significant gene effects of arsenic (+3 oxidation state)-methyltransferase (AS3MT) on MMA%, dimethylarsinic acid percentage (DMA%) and DMA/MMA, purine nucleoside phosphorylase (PNP) on DMA% and glutathione S-transferase omega 2 (GSTO2) on inorganic arsenics (InAs%) were found. Our results show that MMA% might be a potential predictor of cancer risk. The change in MMA% was linked to individual cancer susceptibility related to AS3MT rs3740393.


Rapid evolution and gene expression: a rapidly evolving Mendelian trait that silences field crickets has widespread effects on mRNA and protein expression.
A major advance in modern evolutionary biology is the ability to start linking phenotypic evolution in the wild with genomic changes that underlie that evolution. We capitalized on a rapidly evolving Hawaiian population of crickets (Teleogryllus oceanicus) to test hypotheses about the genomic consequences of a recent Mendelian mutation of large effect which disrupts the development of sound-producing structures on male forewings. The resulting silent phenotype, flatwing, persists because of natural selection imposed by an acoustically orienting parasitoid, but it interferes with mate attraction. We examined gene expression differences in developing wing buds of wild-type and flatwing male crickets using RNA-seq and quantitative proteomics. Most differentially expressed (DE) transcripts were down-regulated in flatwing males (625 up vs. 1716 down), whereas up- and down-regulated proteins were equally represented (30 up and 34 down). Differences between morphs were clearly not restricted to a single pathway, and we recovered annotations associated with a broad array of functions that would not be predicted a priori. Using a candidate gene detection test based on homology, we identified 30% of putative Drosophila wing development genes in the cricket transcriptome, but only 10% were DE. In addition to wing-related annotations, endocrine pathways and several biological processes such as reproduction, immunity and locomotion were DE in the mutant crickets at both biological levels. Our results illuminate the breadth of genetic pathways that are potentially affected in the early stages of adaptation.


Treatment of reperfusion injury in dogs with experimentally induced gastric dilatation-volvulus.
In dogs, gastric dilatation-volvulus (GDV) is characterized by cardiogenic shock, with resulting hypoperfusion. Treatment goals include reperfusion of transiently ischemic tissues, which indicates that reperfusion injury may be a factor in the physiopathogenesis of GDV. Recently, we obtained data that indicate that reperfusion injury may be involved in experimentally induced GDV. Using this GDV model, we evaluated mortality in 24 dogs of 4 equal groups, treated with deferoxamine (an iron chelator), dimethylsulfoxide (a free radical scavenger), a combination of the 2 drugs, or isotonic saline solution. All 6 dogs that were given deferoxamine survived; however, 3 dogs of the dimethylsulfoxide-treated group, 2 dogs of the combination-treated group, and 4 dogs of the saline-treated group died. Results of the study indicate that mortality associated with experimentally induced GDV is reduced by appropriate and timely pharmacologic intervention to prevent or attenuate reperfusion injury, and that deferoxamine may be more effective than dimethylsulfoxide.


Intrafamilial oocyte donation in classic galactosemia: ethical and societal aspects.
Classic galactosemia is a rare inherited disorder of galactose metabolism. Primary ovarian insufficiency (POI) with subfertility affects > 80% of female patients and is an important concern for patients and their parents. Healthcare providers are often consulted for subfertility treatment possibilities. An option brought up by the families is intrafamilial oocyte donation (mother-to-daughter or sister-to-sister). In addition to POI, galactosemia patients can also present varying cognitive and neurological impairments, which may not be fully clear at the time when mother-to-daughter oocyte donation is considered. Ethical and societal aspects arise when exploring this option. This study aimed to provide guidance in aspects to consider based on the views of different groups involved in the oocyte donation process. A qualitative study using in-depth semi-structured interviews with > 50 participants (patients, family members, and healthcare providers) was conducted. From these interviews, themes of concern emerged, which are illustrated and reviewed: (1) family relations, (2) medical impact, (3) patients' cognitive level, (4) agreements to be made in advance and organization of counseling, (5) disclosure to the child, and (6) need for follow-up. We conclude that discussing and carrying out intrafamilial oocyte donation in galactosemia patients requires carefully addressing these themes. This study adds value to the already existing recommendations on intrafamilial oocyte donation in general, since it highlights important additional aspects from the perspectives of patients and their families.


Angiotensin II upregulates type-1 angiotensin II receptors in renal proximal tubule.
Angiotensin II (Ang II) is an important regulator of proximal tubule salt and water reabsorption. Recent studies indicate that rabbit proximal tubule angiotensin II receptors are the type-1 (AT1R) subtype. We studied the effect of Ang II on proximal tubule receptor expression. Rabbits were treated with either angiotensin converting enzyme inhibitors or a low salt diet to modulate endogenous Ang II levels. In captopril-treated rabbits, liver and glomerular AT1R mRNA levels increased 242 +/- 125 and 141 +/- 60%, respectively (n = 6-7; P < 0.05), as determined by quantitative PCR. In contrast, proximal tubule AT1R mRNA levels decreased 40 +/- 11% (n = 6; P < 0.05). Binding of 125I Ang II to renal cortical basolateral membranes of captopril-treated rabbits decreased from 2.9 +/- 0.55 to 1.4 +/- 0.17 fmol/mg protein (n = 6; P < 0.025). In rabbits fed a sodium chloride-deficient diet for 4 wk, AT1R mRNA levels decreased 52 +/- 11% in liver and 43 +/- 7% in glomeruli (n = 4-5; P < 0.05), whereas they increased 141 +/- 85% (n = 5; P < 0.05) in proximal tubule. In basolateral membranes from rabbits on the sodium chloride-deficient diet, specific binding of 125I Ang II increased from 2.1 +/- 0.2 to 4.3 +/- 1.1 fmol/mg protein (n = 7; P < 0.05). To determine whether Ang II directly regulates expression of proximal tubule AT1 receptors, further studies were performed in cultured proximal tubule cells grown from microdissected S1 segments of rabbit proximal tubules and immortalized by transfection with a replication-defective SV40 vector. Incubation of these cells with Ang II (10(-11) to 10(-7) M) led to concentration-dependent increases in both AT1R mRNA levels and specific 125I Ang II binding. Pretreatment with pertussis toxin inhibited Ang II stimulation of AT1R mRNA. AT1R mRNA expression was decreased by either forskolin or a nonhydrolyzable cAMP analogue (dibutryl cAMP). Simultaneous Ang II administration overcame the inhibitory effect of forskolin but not dibutryl cAMP. These results indicate that proximal tubule AT1R expression is regulated by ambient Ang II levels, and Ang II increases AT1R mRNA at least in part by decreasing proximal tubule cAMP generation through a pertussis toxin-sensitive mechanism. Upregulation of proximal tubule AT1R by Ang II may be important in mediating enhanced proximal tubule sodium reabsorption in states of elevated systemic or intrarenal Ang II.


Ten-year stability and latent structure of the DSM-IV schizotypal, borderline, avoidant, and obsessive-compulsive personality disorders.
Evaluation of the validity of personality disorder (PD) diagnostic constructs is important for the impending revision of the Diagnostic and Statistical Manual of Mental Disorders. Prior factor analytic studies have tested these constructs in cross-sectional studies, and models have been replicated longitudinally, but no study has tested a constrained longitudinal model. The authors examined 4 PDs in the Collaborative Longitudinal Personality Disorders study (schizotypal, borderline, avoidant, and obsessive-compulsive) over 7 time points (baseline, 6 months, 1 year, 2 years, 4 years, 6 years, and 10 years). Data for 2-, 4-, 6- and 10-year assessments were obtained in semistructured interviews by raters blind to prior PD diagnoses at each assessment. The latent structure of the 4 constructs was differentiated during the initial time points but became less differentiated over time as the mean levels of the constructs dropped and stability increased. Obsessive-compulsive PD became more correlated with schizotypal and borderline PD than with avoidant PD. The higher correlation among the constructs in later years may reflect greater shared base of pathology for chronic personality disorders.


Predictors of relapse in a prospective study of fluoxetine treatment of major depression.
Loss of response to a previously effective antidepressant is a common clinical problem. Retrospective analyses have shown that the pattern of response during antidepressant treatment (late onset and persistent versus other patterns) can be used to predict relapse during continuation and maintenance treatment and possibly to identify placebo responses to treatment. This study was designed to test the predictive value of response pattern prospectively and to examine the data for other predictors of relapse. Five hundred seventy persons with major depressive disorder were treated with fluoxetine for 12 weeks and their pattern of response was determined. Those who responded (N=292) underwent random assignment, under double-blind conditions, to continue taking fluoxetine or to switch to placebo for 52 weeks or until relapse. Survival analysis was used to examine the effect of covariates on relapse. Although fluoxetine was significantly more effective than placebo during maintenance treatment, this chronically ill group had a high rate of relapse. Contrary to previous findings, a pattern of acute response was not predictive of relapse. Chronicity, symptom severity, a neurovegetative symptom pattern, and female gender were all associated with a significantly greater risk of relapse, with no difference observed between fluoxetine and placebo. The pattern of response to acute treatment appears to be inconsistently predictive of relapse. There is a high rate of relapse with both active medication and placebo in patients with chronic depression. Illness characteristics predict loss of response both to fluoxetine and to placebo. No variable examined was predictive of differential relapse rates between fluoxetine and placebo.


Invasive crayfish impacts on native fish diet and growth vary with fish life stage.
Assessing the impacts of invasive organisms is a major challenge in ecology. Some widespread invasive species such as crayfish are potential competitors and reciprocal predators of ecologically and recreationally important native fish species. Here, we examine the effects of signal crayfish (Pacifastacus leniusculus) on the growth, diet, and trophic position of the chub (Squalius cephalus) in four rivers in Britain. Growth rates of 0+ chub were typically lower in sympatric populations with signal crayfish compared with allopatric populations, and this effect could be traced through to 2+ chub in one river. However, growth rates of older chub (5+ to 6+) were typically higher in the presence of crayfish. Sympatry with crayfish resulted in lower chub length-at-age and mass-at-age in half of the rivers sampled, with no change detected in the other rivers. Stable isotope analyses (δ13C and δ15N) revealed that both chub and crayfish were omnivorous, feeding at multiple trophic levels and occupying similar trophic positions. We found some evidence that chub trophic position was greater at invaded sites on one river, with no difference detected on a second river. Mixing models suggested crayfish were important food items for both small and large chub at invaded sites. This study provides evidence that invasive species can have both positive and negative effects on different life stages of a native species, with the net impact likely to depend on responses at the population level.


Variation in species composition and infection rates of Anopheles mosquitoes at different altitudinal transects, and the risk of malaria in the highland of Dirashe Woreda, south Ethiopia.
The transmission of malaria is heterogeneous, and varies due to altitude. The information on whether the transmission of malaria is indigenous or imported to highland areas is scarce. Therefore, this study aimed to assess the species composition and infection rates of Anopheles at different altitudinal transects, and the risk of malaria if any in the highland of Dirashe Woreda, South Ethiopia. This study was conducted in Gato (low altitude; average elevation of 1273 m), Onota (mid-altitude; average elevation of 1707 m) and Layignaw-Arguba (high altitude; average elevation of 2337 m) from August 2015 to April 2016. Anopheles mosquitoes were sampled using Centers for Disease Control and Prevention (CDC) light traps from thirty houses (ten houses from each village). The circum-sporozoite proteins (CSPs) rate and entomological inoculation rate (EIR) of Anopheles mosquitoes were estimated. For the epidemiological survey, malaria cases were collected from laboratory registration books of selected health facilities from (August 2015-April 2016). A cross-sectional survey was done to collect data on malaria vector control activities in each village (August-September 2015). One thousand two hundred sixty-eight Anopheles mosquitoes comprising Anopheles arabiensis, An. demeilloni, An. cinereus, An. pharoensis, An. funestus-group, An. pretoriensis, An. christyi, An. ardensis and An. tenebrosus were identified in the study area. Anopheles arabiensis was the dominant species in Gato, whereas An. demeilloni was the dominant species in Layignaw-Arguba. Five mosquitoes, three An. arabiensis from Gato and two An. demeilloni from Layignaw-Arguba, were positive for Plasmodium falciparum CSPs. Plasmodium falciparum CSP rate was 0.4% (95% CI: 0.08-1.15) for An. arabiensis in Gato, and it was 0.64% (95% CI: 0.08-2.3) for An. demeilloni from Layignaw-Arguba. The P. falciparum EIR of An. arabiensis was 8.6 (95% CI: 2.4-33.4) infectious bites/person/nine-months in Gato. Plasmodium falciparum parasite was dominant in Gato (88%) and Onota (57.5%), whereas in Layignaw-Arguba P. vivax (59.4%) occurred most frequently. Increased malaria cases were observed in children age 5-14 years in Gato (P < 0.05), whereas in Onota and Layignaw-Arguba there was no statistically significant difference in malaria cases among the age groups. Households owning at least one long lasting insecticidal net were 92.7% in the study area, and 77.6% slept under the net during the preceding night of the survey. About 64.4% of the households in Gato were protected by the indoor residual spray. However, the spraying was done when the density of local malaria vectors was low. Incrimination of Plasmodium CSP positive Anopheles species and the presence of malaria in children under five years in high altitude Layignaw-Arguba may justify the existence of indigenous malaria transmission and the need for effective malaria control. Further investigation and confirmation using more sensitive molecular techniques are however needed to consider An. demeilloni as a proven vector of malaria in Ethiopia.


Effects of inhaled fluticasone on angiogenesis and vascular endothelial growth factor in asthma.
Subepithelial hypervascularity and angiogenesis in the airways are part of structural remodelling of the airway wall in asthma, but the effects of inhaled corticosteroids (ICS) on these have not been explored. Increased vascularity in asthma may contribute to a number of functional abnormalities. A study was undertaken to explore angiogenic modulation by ICS and its likely regulation via vascular endothelial growth factor (VEGF), its receptors and the angiopoietins. A placebo-controlled intervention study with ICS in asthma was performed, examining vascularity, VEGF, its receptors (VEGFR1 and VEGFR2), and angiopoietin-1 (Ang1) to assess which of these factors were changed in the asthmatic airways after ICS treatment. Airway wall biopsy specimens, lavage fluid and cells were obtained from 35 patients with mild asthma randomised to receive ICS or placebo for 3 months, after which bronchoscopic examination and sample collection were repeated. Immunohistochemistry and image analysis were used to obtain quantitative measures of vessels, angiogenic sprouts, VEGF, VEGFR1, VEGFR2 and Ang1 staining in airway biopsy specimens. ELISA was used to assess VEGF concentrations in the lavage fluid. Vessel, VEGF and sprout staining were decreased after 3 months of ICS treatment. VEGF levels remained unchanged. VEGF receptors and Ang1 staining were not reduced after treatment. The findings of this study support an effect of ICS in downregulating angiogenic remodelling in the airways in asthma, associated with decreasing VEGF activity within the airway wall. The environment of the airways after treatment with ICS, with changes in the balance between VEGF, its receptors, Ang1 and sprouts, appears to be less angiogenic than in untreated asthma.


Smooth muscle-derived adventitial progenitor cells direct atherosclerotic plaque composition complexity in a Klf4-dependent manner.
We previously established that vascular smooth muscle-derived adventitial progenitor cells (AdvSca1-SM) preferentially differentiate into myofibroblasts and contribute to fibrosis in response to acute vascular injury. However, the role of these progenitor cells in chronic atherosclerosis has not been defined. Using an AdvSca1-SM cell lineage tracing model, scRNA-Seq, flow cytometry, and histological approaches, we confirmed that AdvSca1-SM-derived cells localized throughout the vessel wall and atherosclerotic plaques, where they primarily differentiated into fibroblasts, smooth muscle cells (SMC), or remained in a stem-like state. Krüppel-like factor 4 (Klf4) knockout specifically in AdvSca1-SM cells induced transition to a more collagen-enriched fibroblast phenotype compared with WT mice. Additionally, Klf4 deletion drastically modified the phenotypes of non-AdvSca1-SM-derived cells, resulting in more contractile SMC and atheroprotective macrophages. Functionally, overall plaque burden was not altered with Klf4 deletion, but multiple indices of plaque composition complexity, including necrotic core area, macrophage accumulation, and fibrous cap thickness, were reduced. Collectively, these data support that modulation of AdvSca1-SM cells through KLF4 depletion confers increased protection from the development of potentially unstable atherosclerotic plaques.


Prospective surveillance of healthcare associated infections in a Cambodian pediatric hospital.
Healthcare associated infections (HAI) are the most common preventable adverse events following admission to healthcare facilities. Data from low-income countries are scarce. We sought to prospectively define HAI incidence at Angkor Hospital for Children (AHC), a Cambodian pediatric referral hospital. Prospective HAI surveillance was introduced for medical admissions to AHC. Cases were identified on daily ward rounds and confirmed using locally adapted Centers for Disease Control and Prevention (CDC) definitions. During the surveillance period, established infection prevention and control (IPC) activities continued, including hand hygiene surveillance. In addition, antimicrobial stewardship practices such as the creation of an antimicrobial guideline smartphone app were introduced. Between 1st January and 31st December 2015 there were 3,263 medical admissions and 102 HAI cases. The incidence of HAI was 4.6/1,000 patient-days (95% confidence interval 3.8-5.6) and rates were highest amongst neonates. Median length of stay was significantly longer in HAI cases: 25 days versus 5 days for non-HAI cases (p < 0.0001). All-cause in-hospital mortality increased from 2.0 to 16.1% with HAI (p < 0.0001). Respiratory infections were the most common HAI (54/102; 52.9%). Amongst culture positive infections, Gram-negative organisms predominated (13/16; 81.3%). Resistance to third generation cephalosporins was common, supporting the use of more expensive carbapenem drugs empirically in HAI cases. The total cost of treatment for all 102 HCAI cases combined, based on additional inpatient days, was estimated to be $299,608. Prospective HAI surveillance can form part of routine practice in low-income healthcare settings. HAI incidence at AHC was relatively low, but human and financial costs remained high due to increased carbapenem use, prolonged admissions and higher mortality rates.


Immunohistochemical visualization of the signature of activated Hedgehog signaling pathway in cutaneous epithelial tumors.
Activation of the Hedgehog (HH) signaling pathway plays a critical role in the development of basal cell carcinoma (BCC). HH signaling activity is produced by nuclear translocation of transcription factors, glioma-associated oncogene homolog (GLI). Among three GLI subfamilies, GLI1 is the only full-length transcriptional activator, and its nuclear localization is recognized as a signature event in HH signaling activation. However, limited published work has investigated the nuclear staining of GLI1 protein in human tumor tissue samples by immunohistochemical analysis. In this study, we performed immunohistochemical staining of GLI1 in 382 cases of cutaneous epithelial tumors, including 196 BCC cases, using rabbit monoclonal antihuman GLI1 antibody (C68H3). As a result, 98.2% cases of BCC showed a diffuse and strong nuclear staining pattern regardless of the histological subtype. Positive staining was mainly restricted to the tumor nests, while the overlying epidermis was negative suggesting specificity of the antibody. In further analysis of other cutaneous epithelial tumors, 100% (4/4) cases of trichoblastoma, 15.1% (5/33) Bowen's disease, 3.5% (1/28) actinic keratosis and 12.5% (4/32) squamous cell carcinoma showed the nuclear staining pattern of GLI1. This suggested that HH signaling is also dysregulated in some other cutaneous malignant tumors. In conclusion, the C68H3 antibody is a useful tool for revealing activation of HH signaling in immunohistochemical analysis.


Structure of sexual networks determines the operation of sexual selection.
Sexual selection is a fundamental evolutionary process but remains debated, particularly in the complexity of polyandrous populations where females mate with multiple males. This lack of resolution is partly because studies have largely ignored the structure of the sexual network, that is, the pattern of mate sharing. Here, we quantify what we call mating assortment with network analysis to specify explicitly the indirect as well as direct relationships between partners. We first review empirical studies, showing that mating assortment varies considerably in nature, due largely to basic properties of the sexual network (size and density) and partly to nonrandom patterns of mate sharing. We then use simulations to show how variation in mating assortment interacts with population-level polyandry to determine the strength of sexual selection on males. Controlling for average polyandry, positive mating assortment, arising when more polygynous males tend to mate with more polyandrous females, drastically decreases the intensity of precopulatory sexual selection on male mating success (Bateman gradient) and the covariance between male mating success and postcopulatory paternity share. Average polyandry independently weakened some measures of sexual selection and crucially also impacted sexual selection indirectly by constraining mating assortment through the saturation of the mating network. Mating assortment therefore represents a key-albeit overlooked-modulator of the strength of sexual selection. Our results show that jointly considering sexual network structure and average polyandry more precisely describes the strength of sexual selection.


Prestin-based outer hair cell motility is necessary for mammalian cochlear amplification.
It is a central tenet of cochlear neurobiology that mammalian ears rely on a local, mechanical amplification process for their high sensitivity and sharp frequency selectivity. While it is generally agreed that outer hair cells provide the amplification, two mechanisms have been proposed: stereociliary motility and somatic motility. The latter is driven by the motor protein prestin. Electrophysiological phenotyping of a prestin knockout mouse intimated that somatic motility is the amplifier. However, outer hair cells of knockout mice have significantly altered mechanical properties, making this mouse model unsatisfactory. Here, we study a mouse model without alteration to outer hair cell and organ of Corti mechanics or to mechanoelectric transduction, but with diminished prestin function. These animals have knockout-like behavior, demonstrating that prestin-based electromotility is required for cochlear amplification.


Stem-cell derived neurosphere assay highlights the effects of viral infection on human cortical development.
Aberrant cortical development is a key feature of neurodevelopmental disorders such as autism spectrum disorder and schizophrenia. Both genetic and environmental risk factors are thought to contribute to defects in cortical development; however, model systems that can capture the dynamic process of human cortical development are not well established. To address this challenge, we combined recent progress in induced pluripotent stem cell differentiation with advanced live cell imaging techniques to establish a novel three-dimensional neurosphere assay, amenable to genetic and environmental modifications, to investigate key aspects of human cortical development in real-time. For the first time, we demonstrate the ability to visualise and quantify radial glial extension and neural migration through live cell imaging. To show proof-of-concept, we used our neurosphere assay to study the effect of a simulated viral infection, a well-established environmental risk factor in neurodevelopmental disorders, on cortical development. This was achieved by exposing neurospheres to the viral mimic, polyinosinic:polycytidylic acid. The results showed significant reductions in radial glia growth and neural migration in three independent differentiations. Further, fixed imaging highlighted reductions in the HOPX-expressing outer radial glia scaffolding and a consequent decrease in the migration of CTIP2-expressing cortical cells. Overall, our results provide new insight into how infections may exert deleterious effects on the developing human cortex.


Expression of MAGE-A12 in oral squamous cell carcinoma.
Melanoma associated-A antigens (MAGE-A) are silent in normal tissues except testis. However, they are activated ina variety of different tumors. Thus, their expression is highly specific to cancer cells. Reverse transcription-nested polymerase chain reaction (RT-nPCR) is a highly sensitive technique that has been used successfully for the detection of MAGE genes in tissue samples. The aim of the study is to analyze the expression rate of MAGE-A12 in oral squamous cell carcinoma (OSCC) using a high sensitive RT-nPCR. Total of 57 tissue samples obtained from patients with OSCC and 20 normal oral mucosal (NOM) probes of otherwise healthy volunteers were included to this study. No expression of MAGE-A12 was observed in the non-neoplasticNOM tissues. MAGE-A12 was expressed in 49.1% of the investigated tumor samples. The correlation between malignant lesion and MAGE-A12 detection was significant (p<0.001). It is concluded that results of this study may indicate MAGE-A12 asa useful additional diagnostic marker especially for the early detection of OSCC distinguishing neoplastic transformation and detection of occult and/or rare disseminated cancer cells. In addition, MAGE-A12 expression in OSCC may also determine anew immunotherapeutic target and might be warranted to develop vaccine for OSCC


Non-esterified fatty acids generate distinct low-molecular weight amyloid-β (Aβ42) oligomers along pathway different from fibril formation.
Amyloid-β (Aβ) peptide aggregation is known to play a central role in the etiology of Alzheimer's disease (AD). Among various aggregates, low-molecular weight soluble oligomers of Aβ are increasingly believed to be the primary neurotoxic agents responsible for memory impairment. Anionic interfaces are known to influence the Aβ aggregation process significantly. Here, we report the effects of interfaces formed by medium-chain (C9-C12), saturated non-esterified fatty acids (NEFAs) on Aβ42 aggregation. NEFAs uniquely affected Aβ42 aggregation rates that depended on both the ratio of Aβ:NEFA as well the critical micelle concentration (CMC) of the NEFAs. More importantly, irrespective of the kind of NEFA used, we observed that two distinct oligomers, 12-18 mers and 4-5 mers were formed via different pathway of aggregation under specific experimental conditions: (i) 12-18 mers were generated near the CMC in which NEFAs augment the rate of Aβ42 aggregation towards fibril formation, and, (ii) 4-5 mers were formed above the CMC, where NEFAs inhibit fibril formation. The data indicated that both 12-18 mers and 4-5 mers are formed along an alternate pathway called 'off-pathway' that did not result in fibril formation and yet have subtle structural and morphological differences that distinguish their bulk molecular behavior. These observations, (i) reflect the possible mechanism of Aβ aggregation in physiological lipid-rich environments, and (ii) reiterate the fact that all oligomeric forms of Aβ need not be obligatory intermediates of the fibril formation pathway.


Pollination-Induced Ethylene in Carnation (Role of Stylar Ethylene in Corolla Senescence).
In carnation (Dianthus caryophyllus L. cv White Sim) cell to cell communication between the pollen and pistil induces ovary development and corolla senescence. The production of elevated ethylene by the style is the first measurable postpollination response. This is followed by a wave of ethylene production from the other floral organs. To investigate the regulation of ethylene biosynthesis in pollinated flowers we measured ethylene production and the expression of 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase transcripts in individual floral organs after pollination. Ethylene production by pollinated styles can be defined temporally by three distinct peaks. By pollinating a single style from a multistyle gynoecium, it was determined that the unpollinated style produces ethylene that corresponds to the first and third peaks observed from a pollinated style. Inhibition of ethylene action in the pollinated style by diazocyclopentadiene treatment prevented both pollination-induced corolla senescence and ethylene production from the ovaries and petals. Treatment with diazocyclopentadiene decreased stylar ethylene production during the second peak and completely inhibited the third peak of ethylene in both pollinated and unpollinated styles. This later auto-catalytic ethylene in styles is likely responsible for pollination-induced corolla senescence and ovary development.


Hyperglycemia induces insulin resistance on angiotensinogen gene expression in diabetic rat kidney proximal tubular cells.
Clinical and animal studies have shown that treatment with angiotensin-converting enzyme (ACE) inhibitors or angiotensin II (Ang II) receptor antagonists slows the progression of nephropathy in diabetes, indicating that Ang II plays an important role in its development. We have reported previously that insulin inhibits the stimulatory effect of high glucose levels on angiotensinogen (ANG) gene expression in rat immortalized renal proximal tubular cells (IRPTCs) via the mitogen-activated protein kinase (p44/42 MAPK) signal transduction pathway. We hypothesize that the suppressive action of insulin on ANG gene expression might be attenuated in renal proximal tubular cells (RPTCs) of rats with established diabetes. Two groups of male adult Wistar rats were studied: controls and streptozotocin (STZ)-induced diabetic rats at 2, 4, 8 and 12 weeks post-STZ administration. Kidney proximal tubules were isolated and cultured in either normal glucose (i.e. 5 mM) or high glucose (i.e. 25 mM) medium to determine the inhibitory effect of insulin on ANG gene expression. Immunoreactive rat ANG (IR-rANG) in culture media and cellular ANG mRNA were measured by a specific radioimmunoassay and reverse transcription-polymerase chain reaction assay respectively. Activation of the p44/42 MAPK signal transduction pathway in rat RPTCs was evaluated by p44/42 MAPK phosphorylation employing a PhosphoPlus p44/42 MAPK antibody kit. Insulin (10(-7) M) inhibited the stimulatory effect of high glucose levels on IR-rANG secretion and ANG gene expression and increased p44/42 MAPK phosphorylation in normal rat RPTCs. In contrast, it failed to affect these parameters in diabetic rat RPTCs. In conclusion, our studies demonstrate that hyperglycaemia induces insulin resistance on ANG gene expression in diabetic rat RPTCs by altering the MAPK signal transduction pathway.


[Acute myocardial infarction after discontinuing aspirin two years after implantation of a drug-eluting coronary stent].
A 48-year-old man was admitted to our cardiac catheterization unit with severe chest pain 75 minutes after onset of symptoms. Two years before he had undergone percutaneous coronary intervention (PCI) for stable angina pectoris with implantation of a drug-eluting stent (TAXUS) into the proximal left anterior descending artery. Antiplatelet therapy with 75 mg clopidogrel was given for one year, together with 100 mg aspirin. Subsequently he was put on low-dose aspirin monotherapy. Eight days before admission aspirin was discontinued because a tooth extraction was planned. The ECG showed significant ST-segment elevation in the precordial leads V1-5. 90 minutes after onset of symptoms coronary angiography was performed and an in-stent thrombosis of the proximal left coronary artery was diagnosed. A successful PCI was performed and abiximab given. The creatine kinase concentration increased to a maximum of 3170 U/l. The pre-discharge echocardiogram showed a slightly reduced left ventricular ejection fraction. After the procedure the patient was stable and free of chest pain and he was discharged from the hospital after one week. Discontinuing antiplatelet therapy, even years after implantation of a drug-eluting coronary stent, increases the risk of a late stent thrombosis. This should be taken into account especially before any procedure, even with a low bleeding risk such as tooth extractions. Antiplatelet treatment should be continued, even if there is a risk increasing minor bleeding complications, so that any life-threatening complication of an acute myocardial infarction is avoided.


Maternal anxiety during pregnancy and newborn epigenome-wide DNA methylation.
Maternal anxiety during pregnancy is associated with adverse foetal, neonatal, and child outcomes, but biological mechanisms remain unclear. Altered foetal DNA methylation (DNAm) has been proposed as a potential underlying mechanism. In the current study, we performed a meta-analysis to examine the associations between maternal anxiety, measured prospectively during pregnancy, and genome-wide DNAm from umbilical cord blood. Sixteen non-overlapping cohorts from 12 independent longitudinal studies of the Pregnancy And Childhood Epigenetics Consortium participated, resulting in a combined dataset of 7243 mother-child dyads. We examined prenatal anxiety in relation to genome-wide DNAm and differentially methylated regions. We observed no association between the general symptoms of anxiety during pregnancy or pregnancy-related anxiety, and DNAm at any of the CpG sites, after multiple-testing correction. Furthermore, we identify no differentially methylated regions associated with maternal anxiety. At the cohort-level, of the 21 associations observed in individual cohorts, none replicated consistently in the other cohorts. In conclusion, contrary to some previous studies proposing cord blood DNAm as a promising potential mechanism explaining the link between maternal anxiety during pregnancy and adverse outcomes in offspring, we found no consistent evidence for any robust associations between maternal anxiety and DNAm in cord blood. Larger studies and analysis of DNAm in other tissues may be needed to establish subtle or subgroup-specific associations between maternal anxiety and the foetal epigenome.


The role of carbohydrate in maintaining extensin in an extended conformation.
Monomers of the plant cell wall glycoprotein extensin are secreted into the wall where they become cross-linked to each other to form a rigid matrix. Expression of the extensin matrix is correlated with the inhibition of further cell elongation during normal development, with increased resistance to virulent pathogens and with other physiological responses characterized by wall strengthening. Carbohydrates make up about two-thirds of the mass of extensin. Arabinose oligomers linked to hydroxyproline residues represent 95% of the total carbohydrate with the remainder occurring as single residues of galactose linked to some serine residues. Electron microscopy of shadowed extensin shows the glycosylated form to be an easily visualized and highly elongated molecule. In contrast, extensin that has been deglycosylated with anhydrous hydrogen fluoride is difficult to resolve in the EM. Glycosylated extensin elutes from a gel filtration column much more rapidly than does the deglycosylated form, and from this analysis we have calculated respective Stokes' radii of 89 and 11 Angstroms for these molecules. Others have shown that inhibition of extensin glycosylation has no effect on its secretion or insolubilization in the cell wall, but that this extensin cannot inhibit cell elongation. It is likely that carbohydrate moieties keep extensin in an extended conformation and that extensin must be in this conformation to form a cross-linked matrix that can function properly in vivo.


Sequence and functional analyses of Haemophilus spp. genomic islands.
A major part of horizontal gene transfer that contributes to the diversification and adaptation of bacteria is facilitated by genomic islands. The evolution of these islands is poorly understood. Some progress was made with the identification of a set of phylogenetically related genomic islands among the Proteobacteria, recognized from the investigation of the evolutionary origins of a Haemophilus influenzae antibiotic resistance island, namely ICEHin1056. More clarity comes from this comparative analysis of seven complete sequences of the ICEHin1056 genomic island subfamily. These genomic islands have core and accessory genes in approximately equal proportion, with none demonstrating recent acquisition from other islands. The number of variable sites within core genes is similar to that found in the host bacteria. Furthermore, the GC content of the core genes is similar to that of the host bacteria (38% to 40%). Most of the core gene content is formed by the syntenic type IV secretion system dependent conjugative module and replicative module. GC content and lack of variable sites indicate that the antibiotic resistance genes were acquired relatively recently. An analysis of conjugation efficiency and antibiotic susceptibility demonstrates that phenotypic expression of genomic island-borne genes differs between different hosts. Genomic islands of the ICEHin1056 subfamily have a longstanding relationship with H. influenzae and H. parainfluenzae and are co-evolving as semi-autonomous genomes within the 'supragenomes' of their host species. They have promoted bacterial diversity and adaptation through becoming efficient vectors of antibiotic resistance by the recent acquisition of antibiotic resistance transposons.


Sex and Catheter Ablation for Ventricular Tachycardia: An International Ventricular Tachycardia Ablation Center Collaborative Group Study.
Significant differences have been described between women and men regarding presentation, mechanism, and treatment outcome of certain arrhythmias. Previous studies of ventricular tachycardia (VT) ablation have not included sufficient women for meaningful comparison. To compare outcomes between women and men with structural heart disease undergoing VT ablation. Investigator-initiated, multicenter, observational study performed between 2002 and 2013, conducted by the International VT Ablation Center Collaborative Group, consisting of 12 high-volume ablation centers. Consecutive patients with structural heart disease undergoing VT ablation were studied. Structural heart disease was defined as left ventricular ejection fraction less than 55%, hypertrophic cardiomyopathy, or right ventricular cardiomyopathy, with scar confirmed during electroanatomic mapping. Catheter ablation. Ventricular tachycardia-free survival and transplant-free survival were compared between women and men. Cox proportional hazard modeling was performed. Of 2062 patients undergoing ablation, 266 (12.9%) were women. Mean (SD) age was 62.4 (13.3) years and 1095 (53.1%) had ischemic cardiomyopathy. Compared with men, women were younger, with higher left ventricular ejection fraction and less VT storm. Despite this, women had higher rates of 1-year VT recurrence following ablation (30.5% vs 25.3%; P = .03). This difference was only partially explained by higher prevalence of nonischemic cardiomyopathy among women and was actually most pronounced among those with ischemic cardiomyopathy. In 12 high-volume ablation centers, women with structural heart disease have worse VT-free survival following ablation than men. Whether this is owing to differences in referral pattern, arrhythmia substrate, or undertreatment requires further study.


Disparities in Quality of Life by Appalachian-Designation Among Women with Breast Cancer.
Few studies have examined the association of geography and quality of life (QOL) among breast cancer patients, particularly differences between Appalachian and non-Appalachian Kentucky women, which is important given the cancer and socioeconomic disparities present in Appalachia. The purpose of this study was to determine whether women with breast cancer residing in Appalachian Kentucky experience poorer health outcomes in regards to depression, stress, QOL, and spiritual wellbeing, relative to those living in non-Appalachian Kentucky after adjusting for demographic, socioeconomic, and health-related factors. Women, aged 18-79, recruited from the Kentucky Cancer Registry between 2009 and 2013 with an incident, primary breast cancer diagnosis completed a telephone interview within 12 months of diagnosis. In this cross-sectional study, sociodemographic characteristics and mental and physical health status were assessed, including number of comorbid conditions, symptoms of depression and stress, and QOL. Among 1245 women with breast cancer, 334 lived in Appalachia and 911 in non-Appalachian counties of Kentucky. Appalachian breast cancer patients differed from non-Appalachian patients on race, education, income, health insurance status, rurality, smoking, and stage at diagnosis. In unadjusted analysis, Appalachian residence was associated with having significantly more comorbid conditions, more symptoms of stress in the past month, and lower Functional Assessment of Cancer Therapy-Breast scores compared to non-Appalachian residence. However, adjustment for sociodemographic and health-related differences by region appear to explain geographic differences in these poorer QOL indicators for women living in Appalachian Kentucky relative to non-Appalachian Kentucky. Policy-, provider-, and individual-level implications are discussed.


Adult-specific systemic over-expression reveals novel in vivo effects of the soluble forms of ActRIIA, ActRIIB and BMPRII.
Bone morphogenetic proteins (BMPs)/growth differentiation factors (GDFs), which belong to the TGF-beta superfamily, are pleiotropic factors that play a role in regulating the embryonic development and postnatal homeostasis of various organs and tissues by controlling cellular differentiation, proliferation and apoptosis. Conventional transgenic and knockout (KO) mouse approaches have provided only limited information regarding the in vivo functions of BMP signaling in adult animals due to the effects on prenatal development and the difficulty in manipulating multiligand signals simultaneously. We recently produced transgenic chimeric mice(Tg chimeras) in which the soluble IgG1-Fc fusion protein of three BMP type II receptors (ActRIIA, ActRIIB, BMPRII) was highly circulated (281-709 μg/ml), specifically in adult mouse blood. Since each BMP receptor can bind to multiple BMP ligands, these Tg chimeras should be useful to investigate the effects of trapping multiple BMP ligands. Remarkably, some phenotypes were unexpected based on previous studies, such as KO mouse analyses, presumably representing the effects of the multiple ligand trapping. These phenotypes included increased red blood cells (RBCs) and decreased viability in adults. In a further study, we focused on the phenotype of increased RBCs and found that extramedullary hematopoiesis in the spleen, not in the bone marrow, was increased using histological and flow cytometric analyses. Although it remains to be elucidated whether the transgene products affect the tissues directly or indirectly, our data provide novel and important insight into the biological functions of the soluble IgG1-Fc fusion protein of three BMP type II receptors in adults, and our approach should have broad applications to research on other ligand receptor families and studies involving mouse models.


Ocean-wide tracking of pelagic sharks reveals extent of overlap with longline fishing hotspots.
Overfishing is arguably the greatest ecological threat facing the oceans, yet catches of many highly migratory fishes including oceanic sharks remain largely unregulated with poor monitoring and data reporting. Oceanic shark conservation is hampered by basic knowledge gaps about where sharks aggregate across population ranges and precisely where they overlap with fishers. Using satellite tracking data from six shark species across the North Atlantic, we show that pelagic sharks occupy predictable habitat hotspots of high space use. Movement modeling showed sharks preferred habitats characterized by strong sea surface-temperature gradients (fronts) over other available habitats. However, simultaneous Global Positioning System (GPS) tracking of the entire Spanish and Portuguese longline-vessel fishing fleets show an 80% overlap of fished areas with hotspots, potentially increasing shark susceptibility to fishing exploitation. Regions of high overlap between oceanic tagged sharks and longliners included the North Atlantic Current/Labrador Current convergence zone and the Mid-Atlantic Ridge southwest of the Azores. In these main regions, and subareas within them, shark/vessel co-occurrence was spatially and temporally persistent between years, highlighting how broadly the fishing exploitation efficiently "tracks" oceanic sharks within their space-use hotspots year-round. Given this intense focus of longliners on shark hotspots, our study argues the need for international catch limits for pelagic sharks and identifies a future role of combining fine-scale fish and vessel telemetry to inform the ocean-scale management of fisheries.


Revision of the family Chasmocarcinidae Serène, 1964 (Crustacea, Brachyura, Goneplacoidea).
The family Chasmocarcinidae Serène, 1964, is revised based on the examination of the type material of many of its species as well as unidentified and previously identified material from around the world. The revised family now consists of three subfamilies comprising 16 genera (including eight described as new) and 51 species (including 19 described as new). The subfamily Chasmocarciinae Serène, 1964, consists of Amboplax n. gen. with one species; Angustopelta n. gen. with four species, two of which are new; Camatopsis Alcock & Anderson, 1899, with six species, five of which are new; Chasmocarcinops Alcock, 1900, with one species; Chasmocarcinus Rathbun, 1898, with 11 species, one of which is new; Chinommatia n. gen. with five species, two of which are new; Deltopelta n. gen. with one species; Hephthopelta Alcock, 1899, with two species, one of which is new; Microtopsis Komai, Ng & Yamada, 2012, with two species, one of which is new; Notopelta n. gen. with one species; Statommatia n. gen. with five species, two of which are new; and Tenagopelta n. gen. with three species, two of which are new. The subfamily Megaesthesiinae Števčić, 2005, consists of Alainthesius n. gen. with two species, both of which are new; Megaesthesius Rathbun, 1909, with four species, one of which is new. The subfamily Trogloplacinae Guinot, 1986, consists of Australocarcinus Davie, 1988, with three species, and Trogloplax Guinot, 1986, with one species. A neotype is selected for Chasmocarcinus cylindricus Rathbun, 1901. Three nominal species were found to be junior subjective synonyms of other species: Chasmocarcinus panamensis Serène, 1964, of C. longipes Garth, 1940; Chasmocarcinus rathbuni Bouvier, 1917, of C. typicus Rathbun, 1898; and Hephthopelta superba Boone, 1927, of Deltopelta obliqua (Rathbun, 1898). Thirteen chasmocarcinid genera are exclusively found in the Indo-West Pacific region, one (Chasmocarcinus) in both the Western Atlantic and Tropical Eastern Pacific regions, and two (Deltopelta n. gen. and Amboplax n. gen.) exclusively in the Western Atlantic. Chasmocarcinids are remarkable for occurring from depths exceeding 1000 m to shallow water and completely freshwater habitats: chasmocarcinines and megaesthesiines are found from shallow to deep water marine ecosystems, whereas trogloplacines live in freshwater streams, including cave systems.


Self-perception of School-aged Children With Amblyopia and Its Association With Reading Speed and Motor Skills.
Reading and eye-hand coordination deficits in children with amblyopia may impede their ability to demonstrate their knowledge and skills, compete in sports and physical activities, and interact with peers. Because perceived scholastic, social, and athletic competence are key determinants of self-esteem in school-aged children, these deficits may influence a child's self-perception. To determine whether amblyopia is associated with lowered self-perception of competence, appearance, conduct, and global self-worth and whether the self-perception of children with amblyopia is associated with their performance of reading and eye-hand tasks. This cross-sectional study was conducted from January 2016 to June 2017 at the Pediatric Vision Laboratory of the Retina Foundation of the Southwest and included healthy children in grades 3 to 8, including 50 children with amblyopia; 13 children without amblyopia with strabismus, anisometropia, or both; and 18 control children. Self-perception was assessed using the Self-perception Profile for Children, which includes 5 domains: scholastic, social, and athletic competence; physical appearance; behavioral conduct; and a separate scale for global self-worth. Reading speed and eye-hand task performance were evaluated with the Readalyzer (Bernell) and Movement Assessment Battery for Children, 2nd Edition. Visual acuity and stereoacuity also were assessed. Of 50 participants, 31 (62%) were girls, 31 (62%) were non-Hispanic white, 6 (12%) were Hispanic white, 3 (6%) were African American, 4 (8%) were Asian/Pacific Islander, and 3 (6%) were more than 1 race/ethnicity, and the mean [SD] age was 10.6 [1.3] years. Children with amblyopia had significantly lower scores than control children for scholastic (mean [SD], 2.93 [0.74] vs 3.58 [0.24]; mean [SD] difference, 0.65 [0.36]; 95% CI, 0.29-1.01; P = .004), social (mean [SD], 2.95 [0.64] vs 3.62 [0.35]; mean [SD] difference, 0.67 [0.32]; 95% CI, 0.35-0.99] P < .001), and athletic (mean [SD], 2.61 [0.65] vs 3.43 [0.52]; mean [SD] difference, 0.82 [0.34]; 95% CI, 0.48-1.16; P = .001) competence domains. Among children with amblyopia, a lower self-perception of scholastic competence was associated with a slower reading speed (r = 0.49, 95% CI, 0.17-0.72; P = .002) and a lower self-perception of scholastic, social, and athletic competence was associated with worse performance of aiming and catching (scholastic r = 0.48; 95% CI, 0.16-0.71; P = .007; social r = 0.63; 95% CI, 0.35-0.81; P < .001; athletic r = 0.53; 95% CI, 0.21-0.75; P = .003). No differences in the self-perception of physical appearance (mean [SD], 3.32 [0.63] vs 3.64 [0.40]), conduct (mean [SD], 3.09 [0.56] vs 3.34 [0.66]), or global self-worth (mean [SD], 3.42 [0.42] vs 3.69 [0.36]) were found between the amblyopic and control groups. These findings suggest that lower self-perception is associated with slower reading speed and worse motor skills and may highlight the wide-ranging effects of altered visual development for children with amblyopia in their everyday lives.


Epstein-Barr virus nuclear antigen 1 interacts with regulator of chromosome condensation 1 dynamically throughout the cell cycle.
The Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is a sequence-specific DNA-binding protein that plays an essential role in viral episome replication and segregation, by recruiting the cellular complex of DNA replication onto the origin (oriP) and by tethering the viral DNA onto the mitotic chromosomes. Whereas the mechanisms of viral DNA replication are well documented, those involved in tethering EBNA1 to the cellular chromatin are far from being understood. Here, we have identified regulator of chromosome condensation 1 (RCC1) as a novel cellular partner for EBNA1. RCC1 is the major nuclear guanine nucleotide exchange factor for the small GTPase Ran enzyme. RCC1, associated with chromatin, is involved in the formation of RanGTP gradients critical for nucleo-cytoplasmic transport, mitotic spindle formation and nuclear envelope reassembly following mitosis. Using several approaches, we have demonstrated a direct interaction between these two proteins and found that the EBNA1 domains responsible for EBNA1 tethering to the mitotic chromosomes are also involved in the interaction with RCC1. The use of an EBNA1 peptide array confirmed the interaction of RCC1 with these regions and also the importance of the N-terminal region of RCC1 in this interaction. Finally, using confocal microscopy and Förster resonance energy transfer analysis to follow the dynamics of interaction between the two proteins throughout the cell cycle, we have demonstrated that EBNA1 and RCC1 closely associate on the chromosomes during metaphase, suggesting an essential role for the interaction during this phase, perhaps in tethering EBNA1 to mitotic chromosomes.


Peripheral chemoreceptor control of exercise hyperpnea in humans.
Estimates of the proportional contribution of the peripheral chemoreceptors (i.e., the carotid bodies) to human ventilatory control during moderate exercise (i.e., below the lactate threshold, theta L) suggest that they: (a) exert no discernible influence on the initial (usually rapid) phase I component; (b) provide significant modulation of the slower, exponential phase II dynamics, therefore contributing to the tightness of arterial PCO2 regulation and the magnitude of the transient hypoxemia in this phase; and (c) account for approximately 20% of the steady-state phase III drive, which can rise to over 50% in hypoxia (PaO2 approximately 50 mm Hg). Above theta L, the carotid bodies constrain the transient fall in arterial pH by mediating much (but not all) of the compensatory hyperventilation for the metabolic acidemia. The carotid body contribution above theta L, estimated by Dejours O2 testing, is not appreciably different from subthreshold estimates, suggesting that: (a) the respiratory alkalosis in blood and cerebrospinal fluid resulting from the hyperventilation may suppress carotid chemosensitivity; (b) an artifact resulting from secondary hyperoxia-induced stimulation of central chemoreceptors may lead to underestimation of the carotid body contribution; or (c) the carotid bodies may not be entirely "silenced" by hyperoxia during a metabolic acidemia.


Beta-blockers and depression: the more the murkier?
To review the literature regarding the purported association between oral ingestion of beta-blocker drugs and depressed mood. MEDLINE was searched for published articles using the key words propranolol, atenolol, metoprolol, nadolol, timolol, beta-blocker, beta-adrenergic antagonist, or beta-adrenergic blocker in combination with the key words depression, depressive symptomatology, major depressive disorder, or depressed mood from January 1966 through December 1996. Findings regarding the association are equivocal. Plausible explanations include study design, case definition, and confounding disease states. Most of the evidence supporting an association has used case series and case reports. Findings from cross-sectional observational studies and case-control studies are equivocal. Case definition and measurement instruments may partially explain these inconsistencies. Studies using a diagnosis of depression generally do not support the relationship. Trials using depressive symptoms are about evenly split, but they have generally enrolled a small number of patients and have questionable statistical power. Studies defining antidepressant prescriptions dispensed as a marker for depression generally support the association. Evidence exists both for and against the hypothesis that lipophilic beta-blockers cause more depression than do hydrophilic beta-blockers. beta-Blockers may have been unjustly associated with depression and their use avoided for that reason. Future studies into the association between depression and beta-blocker use should evaluate whether the association is affected by case definition and study design characteristics, including disease, dose-response, bias, measurement error, or ability to precisely measure the length of the exposure.


Unusual active site location and catalytic apparatus in a glycoside hydrolase family.
The human gut microbiota use complex carbohydrates as major nutrients. The requirement for an efficient glycan degrading systems exerts a major selection pressure on this microbial community. Thus, we propose that these bacteria represent a substantial resource for discovering novel carbohydrate active enzymes. To test this hypothesis, we focused on enzymes that hydrolyze rhamnosidic bonds, as cleavage of these linkages is chemically challenging and there is a paucity of information on l-rhamnosidases. Here we screened the activity of enzymes derived from the human gut microbiota bacterium Bacteroides thetaiotaomicron, which are up-regulated in response to rhamnose-containing glycans. We identified an α-l-rhamnosidase, BT3686, which is the founding member of a glycoside hydrolase (GH) family, GH145. In contrast to other rhamnosidases, BT3686 cleaved l-Rha-α1,4-d-GlcA linkages through a retaining double-displacement mechanism. The crystal structure of BT3686 showed that the enzyme displayed a type A seven-bladed β-propeller fold. Mutagenesis and crystallographic studies, including the structure of BT3686 in complex with the reaction product GlcA, revealed a location for the active site among β-propeller enzymes cited on the posterior surface of the rhamnosidase. In contrast to the vast majority of GH, the catalytic apparatus of BT3686 does not comprise a pair of carboxylic acid residues but, uniquely, a single histidine functions as the only discernable catalytic amino acid. Intriguingly, the histidine, His48, is not invariant in GH145; however, when engineered into structural homologs lacking the imidazole residue, α-l-rhamnosidase activity was established. The potential contribution of His48 to the catalytic activity of BT3686 is discussed.


Home, training and work: mobility of British doctors.
This study examines the locations of family homes, medical schools and places of specialist training, and work of doctors qualifying from UK medical schools in 5 calendar years between 1974 and 1993. The contribution of each UK region to the medical workforce relative to its population is assessed and trends over time are examined. The relationship between place of family home and medical school attended is examined for 14,108 doctors. Career appointment location and its relationship to medical school and family home location are examined for over 4000 doctors. For the qualifiers of 1983, an additional analysis incorporating place of training is included. Large differences were found in the percentage of medical students from local family homes attending each regional medical school. In some cases differences reflected local populations but other cases had no obvious cause. Over all cohorts studied, 38% of respondents attended a medical school in the region of their family home (32% of 1993 qualifiers), 42% held a career post in the same region as their medical school, and 38% held a career post in the same region as their family home. Among the qualifiers of 1983, 65% had a career post in the same region as their postgraduate training, 34% also attended medical school in the same region, and 19% also came from family homes in the same region. More women than men took up a career post in the same region as their postgraduate training. The relationships to family home and medical school did not differ by gender. Consultants appeared slightly less likely than GPs to have stayed within a region, but this difference was not statistically significant.