Coffee consumption and reduced risk of hepatocellular carcinoma: findings from the Singapore Chinese Health Study.
Coffee consumption has been associated with reduced markers of hepatic cell damage, reduced risk of chronic liver disease, and cirrhosis across a variety of populations. Data on the association between coffee consumption and risk of hepatocellular carcinoma (HCC), especially in high-risk populations, are sparse. This study examines the relationship between coffee and caffeine consumption, and the risk of developing HCC within the Singapore Chinese Health Study, a prospective cohort of 63,257 middle-aged and older Chinese men and women, a relatively high-risk population for HCC. Baseline data on coffee consumption and other dietary and lifestyle factors were collected through in-person interviews at enrollment between 1993 and 1998. As of 31 December 2006, 362 cohort participants had developed HCC. High levels of coffee or caffeine consumption were associated with reduced risk of HCC (p for trend < 0.05). Compared with non-drinkers of coffee, individuals who consumed three or more cups of coffee per day experienced a statistically significant 44% reduction in risk of HCC (hazard ratio 0.56, 95% confidence interval, 0.31-1.00, p = .049) after adjustment for potential confounders and tea consumption. These data suggest that coffee consumption may reduce the risk of developing HCC in Chinese in Singapore.


No indication of strict host associations in a widespread mycoparasite: grapevine powdery mildew (Erysiphe necator) is attacked by phylogenetically distant Ampelomyces strains in the field.
Pycnidial fungi belonging to the genus Ampelomyces are common intracellular mycoparasites of powdery mildews worldwide. Some strains have already been developed as commercial biocontrol agents (BCAs) of Erysiphe necator and other powdery mildew species infecting important crops. One of the basic, and still debated, questions concerning the tritrophic relationships between host plants, powdery mildew fungi, and Ampelomyces mycoparasites is whether Ampelomyces strains isolated from certain species of the Erysiphales are narrowly specialized to their original mycohosts or are generalist mycoparasites of many powdery mildew fungi. This is also important for the use of Ampelomyces strains as BCAs. To understand this relationship, the nuclear ribosomal DNA internal transcribed spacer (ITS) and partial actin gene (act1) sequences of 55 Ampelomyces strains from E. necator were analyzed together with those of 47 strains isolated from other powdery mildew species. These phylogenetic analyses distinguished five major clades and strains from E. necator that were present in all but one clade. This work was supplemented with the selection of nine inter-simple sequence repeat (ISSR) markers for strain-specific identification of Ampelomyces mycoparasites to monitor the environmental fate of strains applied as BCAs. The genetic distances among strains calculated based on ISSR patterns have also highlighted the genetic diversity of Ampelomyces mycoparasites naturally occurring in grapevine powdery mildew. Overall, this work showed that Ampelomyces strains isolated from E. necator are genetically diverse and there is no indication of strict mycohost associations in these strains. However, these results cannot rule out a certain degree of quantitative association between at least some of the Ampelomyces lineages identified in this work and their original mycohosts.


High Prevalence of Syphilis and Inadequate Prenatal Care in Brazilian Pregnant Women: A Cross-Sectional Study.
In Brazil, the reported number of syphilis cases among pregnant women has markedly increased. We conducted a cross-sectional study to determine the prevalence of Treponema pallidum infection and associated factors in pregnant women in Dourados, Mato Grosso do Sul, Brazil. Participants voluntarily completed a risk-factor questionnaire and provided blood specimens for unlinked anonymous testing for the presence of antibodies against T. pallidum, the causative agent of syphilis. Data of newborns were obtained from medical records. We performed univariate and multivariate regression analyses to assess associations with syphilis. The seroprevalence of syphilis in pregnant women was 4.4% (n = 29/661). Twenty-five newborns were seropositive for T. pallidum, and complications due to syphilis were observed in 28% (n = 7/25). Although 96.5% (n = 28/29) of women with syphilis received antenatal care, Venereal Disease Research Laboratory tests were performed in the first trimester for 47.6% (n = 10/21) of women. Women who received treatment in the third trimester (28.6%; n = 6/21) were considered successfully treated at the time of delivery. The use of illicit drugs during pregnancy (odds ratio [OR]: 13.3, 95% CI: 1.9-91.2) and a history of abortion (OR: 3.7, 95% CI: 1.7-8) were associated with syphilis. Our findings highlight that the poor quality of antenatal care services contributes to the high prevalence of syphilis. In addition, there are social and behavioral risk factors associated with syphilis in pregnant women. Future studies are needed to determine limitations of clinical management and control services available to pregnant women with syphilis.


Pattern formation in the striatum: neurons with early projections to the substantia nigra survive the cell death period.
During the early postnatal period the striatum undergoes significant cell death. The specificity and regulation of this regressive event may be particularly interesting in the light of recent findings demonstrating that a developmentally organized compartmental architecture exists in the striatum. The striatum can be divided into two complementary and phenotypically distinct compartments, the patches and the matrix. In the adult, these two striatal compartments can be distinguished on the basis of their anatomy and a series of compartment-specific biochemical and hodological markers. We have previously demonstrated that the neurons within the patch and matrix compartments become postmitotic and make connections with the substantia nigra at distinct and sequential developmental times. The majority of patch neurons become postmitotic between embryonic days 12 and 15 and make a striatonigral connection prenatally. In contrast, a majority of matrix neurons become postmitotic between embryonic days 17 and 20 and do not form an efferent connection to the substantia nigra until the first postnatal week. Here we investigated whether either neuronal birthdate or time of making an efferent projection correlates with a neuron's probability of surviving the cell death period. We found that both the patch and matrix compartments undergo their entire cell death period by the end of the first postnatal week. During this period approximately 30% of striatal neurons are subject to cell death, regardless of striatal compartment. Neuronal counts within the striatal patch compartment suggest that both early born neurons (embryonic day 13) and early projecting neurons (to the substantia nigra) are preferentially spared. However, their considerable overlap (i.e., most early born neurons also have a nigral projection) prevents assessment of which feature is critical for survival. In contrast, there are small, but mostly separate, populations of early born and early projecting neurons within the matrix compartment. Quantitative analysis of these two distinct populations suggests that while early projection neurons within the matrix are spared, the early born matrix neurons lacking an early nigral projection undergo significant cell death. This proposal is further supported by the observation that the percentage of early born neurons in both the patch and matrix compartments that also have an early nigral projection increases from postnatal day 2 to 17. This finding suggests that among the early born striatal neurons in both compartments, those that do not project to the nigra selectively die during the cell death period. Together these results support the hypothesis that completion of an early projection to the substantia nigra gives neurons an advantage for surviving the cell death period.


The natural history of type II endoleaks after endovascular aneurysm repair for ruptured abdominal aortic aneurysm.
The natural history of type II endoleak (T2EL) after endovascular aneurysm repair (EVAR) for ruptured abdominal aortic aneurysm (rAAA) remains elusive; as such, treatment guidelines are ill defined. The purpose of this study was to better delineate the natural history of T2EL after EVAR for rAAA in an effort to determine the need for reintervention and optimal surveillance. A retrospective record review was conducted of all patients undergoing EVAR for rAAA in two large tertiary care academic vascular centers. Patient demographics, comorbidities, anatomic variables, and operative details were analyzed. Primary outcomes included the presence of T2EL, reintervention, delayed rupture, and aneurysm-related death. EVAR was used to treat rAAA in 56 patients between 2000 and 2013. Mean follow-up of this cohort was 634 days. Completion arteriogram demonstrated T2ELs in 12 patients (21%), and an additional four T2ELs (7%) were found by postoperative computed tomography angiogram that were not identified on the completion angiogram. Body mass index was the only statistically significant variable associated with the development of T2EL (P = .03). Preoperative warfarin use, aortic thrombus burden, and device type did not correlate with T2EL development. Iliolumbar vessels supplied 75% (n = 12) of T2ELs. Of the 14 patients with T2ELs who underwent serial imaging postoperatively, six (43%) sealed spontaneously. Five patients (36%) underwent reintervention for T2EL by way of coil embolization-four in which treatment was initiated by attending preference. One patient was treated for ongoing anemia in the immediate postoperative period. There was no sac expansion, delayed rupture, or graft explantation. T2ELs after EVAR for rAAA are common and appear to be associated with a benign natural history if left untreated. Although many will spontaneously seal early in the postoperative period, those that remain patent do not appear to increase the risk for sac expansion or delayed rupture or affect patient survival. As such, a conservative approach to treatment of T2ELs in rAAA may be warranted.


Conditional cooperation in group contests.
In this paper we show experimentally that conditional cooperation, a phenomenon described in the private provision of public goods, is also present in group contests, where participants' contributions to their group performance partially determines if they overcome a rival group. This environment allows us to identify new determinants of conditional cooperation. We observe conditional cooperation in successful groups and in groups where members contribute more than rivals (even if they lose), but it vanishes in those groups that lose the contest due to low group performance. A random-effect linear panel regression analysis with an extensive set of controls confirms the findings.


Free-electron laser etching of dental enamel.
The purpose of this study was to evaluate the Mark-III free-electron laser as a means of etching enamel surfaces, with potential application to resin bonding. The FEL was tuned to wavelengths ranging from 3.0 to 9.2 microm. Specific wavelengths that are resonantly absorbed by phosphates, proteins, and water were used. First, bovine enamel was polished and exposed to static FEL exposures. Lased enamel was examined using scanning electron microscopy (SEM). Additional bovine enamel specimens were exposed to FEL at similar wavelengths, but with rastering to create treated rectangular areas on each specimen. Surface roughness was evaluated using profilometry and atomic force microscopy (AFM). Composite was bonded to the lased enamel, and shear bond strengths were determined using an Instron universal testing machine. As a control, the surface roughness of, and shear bond strengths to, acid-etched enamel were determined. Static FEL exposures caused changes in the enamel ranging from an etched appearance to pits, cracks, and frank cratering. The surface roughness of lased enamel was much greater than that of acid-etched enamel, and was qualitatively different as well. Shear bond strengths of resin to acid-etched enamel were significantly higher than bond strengths to lased enamel. Under the conditions used in this study, the FEL did not offer a practical and effective method of etching enamel for resin bonding. However, the ability of the FEL to deliver many specific wavelengths makes it an interesting tool for further research of laser effects on tooth structure.


Resonantly pumped high efficiency Ho:YAG laser.
High-efficient CW and Q-switched Ho:YAG lasers resonantly dual-end-pumped by two diode-pumped Tm:YLF lasers at 1908 nm were investigated. A maximum slope efficiency of 74.8% in CW operation as well as a maximum output power of 58.7 W at 83.2 W incident pump power was achieved, which corresponded to an optical-to-optical conversion efficiency of 70.6%. The maximum pulse energy of 2.94 mJ was achieved, with a 31 ns FWHM pulse width and a peak power of approximately 94.7 kW.


Transparency in conducting and reporting research: A survey of authors, reviewers, and editors across scholarly disciplines.
Calls have been made for improving transparency in conducting and reporting research, improving work climates, and preventing detrimental research practices. To assess attitudes and practices regarding these topics, we sent a survey to authors, reviewers, and editors. We received 3,659 (4.9%) responses out of 74,749 delivered emails. We found no significant differences between authors', reviewers', and editors' attitudes towards transparency in conducting and reporting research, or towards their perceptions of work climates. Undeserved authorship was perceived by all groups as the most prevalent detrimental research practice, while fabrication, falsification, plagiarism, and not citing prior relevant research, were seen as more prevalent by editors than authors or reviewers. Overall, 20% of respondents admitted sacrificing the quality of their publications for quantity, and 14% reported that funders interfered in their study design or reporting. While survey respondents came from 126 different countries, due to the survey's overall low response rate our results might not necessarily be generalizable. Nevertheless, results indicate that greater involvement of all stakeholders is needed to align actual practices with current recommendations.


Recent explosive human population growth has resulted in an excess of rare genetic variants.
Human populations have experienced recent explosive growth, expanding by at least three orders of magnitude over the past 400 generations. This departure from equilibrium skews patterns of genetic variation and distorts basic principles of population genetics. We characterized the empirical signatures of explosive growth on the site frequency spectrum and found that the discrepancy in rare variant abundance across demographic modeling studies is mostly due to differences in sample size. Rapid recent growth increases the load of rare variants and is likely to play a role in the individual genetic burden of complex disease risk. Hence, the extreme recent human population growth needs to be taken into consideration in studying the genetics of complex diseases and traits.


Inactivation of TNF signaling by rationally designed dominant-negative TNF variants.
Tumor necrosis factor (TNF) is a key regulator of inflammatory responses and has been implicated in many pathological conditions. We used structure-based design to engineer variant TNF proteins that rapidly form heterotrimers with native TNF to give complexes that neither bind to nor stimulate signaling through TNF receptors. Thus, TNF is inactivated by sequestration. Dominant-negative TNFs represent a possible approach to anti-inflammatory biotherapeutics, and experiments in animal models show that the strategy can attenuate TNF-mediated pathology. Similar rational design could be used to engineer inhibitors of additional TNF superfamily cytokines as well as other multimeric ligands.


Frequency-specific relation between hippocampal theta rhythm, behavior, and amobarbital action.
The frequency of the hippocampal theta rhythm in freely moving rats varies predictably in relation to behavior in a simple learning situation. The theta rhythm may be driven by electrical stimulation of the medial septal area at frequencies within the theta range. The threshold for septal driving is lowest at that frequency which the rat displays in response to frustrative nonreward; the driving threshold is selectively raised at this frequency by sodium amobarbital. It is suggested that the behavioral effects of amobarbital are due to a disruption of the theta frequency normally displayed in response to nonreward.


The actin cable is dispensable in directing dorsal closure dynamics but neutralizes mechanical stress to prevent scarring in the Drosophila embryo.
The actin cable is a supracellular structure that embryonic epithelia produce to close gaps. However, the action of the cable remains debated. Here, we address the function of the cable using Drosophila dorsal closure, a paradigm to understand wound healing. First, we show that the actin cytoskeleton protein Zasp52 is specifically required for actin cable formation. Next, we used Zasp52 loss of function to dissect the mechanism of action of the cable. Surprisingly, closure dynamics are perfect in Zasp52 mutants: the cable is therefore dispensable for closure, even in the absence of the amnioserosa. Conversely, we observed that the cable protects cellular geometries from robust morphogenetic forces that otherwise interfere with closure: the absence of cable results in defects in epithelial organization that lead to morphogenetic scarring. We propose that the cable prevents morphogenetic scarring by stabilizing cellular interactions rather than by acting on closure dynamics.


Religious Involvement and Telomere Length in Women Family Caregivers.
Telomere length (TL) is an indicator of cellular aging associated with longevity and psychosocial stress. We examine here the relationship between religious involvement and TL in 251 stressed female family caregivers recruited into a 2-site study. Religious involvement, perceived stress, caregiver burden, depressive symptoms, and social support were measured and correlated with TL in whole blood leukocytes. Results indicated a U-shaped relationship between religiosity and TL. Those scoring in the lowest 10% on religiosity tended to have the longest telomeres (5743 bp ± 367 vs. 5595 ± 383, p = 0.069). However, among the 90% of caregivers who were at least somewhat religious, religiosity was significantly and positively related to TL after controlling for covariates (B = 1.74, SE = 0.82, p = 0.034). Whereas nonreligious caregivers have relatively long telomeres, we found a positive relationship between religiosity and TL among those who are at least somewhat religious.


Expanding the use of simulation in open vascular surgical training.
Simulation technology has a well-defined role in nonmedical professions such as aviation and over the last two decades has permeated medical training. The most successful surgical simulation is in the fields of laparoscopic and endovascular surgery. These two-dimensional scenarios, as in the aviation industry, lend themselves to simulation. Open simulators have been met with more resistance than their laparoscopic counterparts because of the difficulties in simulating the three-dimensional field. Engaging in persistent practice is what makes the expert and all trainees should aspire to this. Without knowing, all surgical trainees have engaged in deliberate practice when first learning to tie surgical knots. This deliberate practice should be used in all aspects of vascular surgical practice, and it is no longer acceptable to perform procedures such as arterial anastomoses for the first time on patients. Simulators exist for all aspects of vascular surgical training and vary in complexity and price. Some of these simulators are suitable for use at home or in a skills laboratory whereas others are more suitable for use in a specialized skills center. Training on these simulators can be offered at a local level or at a regional level in the skills center. Where surgical procedures are not commonly performed or expertise is required for a new innovation, it is more appropriate to have national or internationally based workshops under the auspices of surgical boards or societies. Simulation of crisis management, well known in aviation, has also been applied to vascular surgical practice and can offer benefit to senior trainees even when their performance on a noncrisis simulator has reached a plateau. This article identifies the areas where simulation in open vascular surgery can benefit the trainee.


Is Dementia in Parkinson' Disease Related to Chronic Stress, Anxiety, and Depression?
Stress, anxiety, and depression are known to be associated with the development of neurodegenerative disorders through interactions with the underlying pathophysiology. We hypothesized that the presence of these symptoms contributes to cognitive disturbances and dementia in Parkinson's disease (PD). The present study aimed to investigate the levels of stress, anxiety, and depression in PD patients relative to healthy individuals. Anxiety, stress, and depression levels were assessed using standardized questionnaires in PD without dementia (PDND, n = 30), PD with dementia (PDD, n = 28), and healthy controls (HC, n = 26). Arithmetic subtraction task was used as a stressor. Galvanic skin response, heart rate and salivary cortisol, and alpha-amylase were measured during baseline and after induced stress (arithmetic task). Acute anxiety, acute stress, and depression levels were significantly higher in PDND compared to HC, whereas both acute and chronic anxiety, stress, and depression levels were significantly higher in PDD compared to PDND and HC. Cortisol and alpha-amylase levels were significantly higher in PDND compared to HC during both baseline and postarithmetic task. Posttask levels of cortisol were lower in PDD compared to PDND. This study concludes that higher levels of salivary cortisol and alpha-amylase at baseline and poststress task with normal levels of chronic stress and anxiety were associated with no dementia in PD. Presence of higher levels of acute, chronic anxiety, and stress along with depression with lower cortisol reactivity to stressor suggests onset of dementia in Parkinson's patients.


Structures of the human GTPase MMAA and vitamin B12-dependent methylmalonyl-CoA mutase and insight into their complex formation.
Vitamin B(12) (cobalamin, Cbl) is essential to the function of two human enzymes, methionine synthase (MS) and methylmalonyl-CoA mutase (MUT). The conversion of dietary Cbl to its cofactor forms, methyl-Cbl (MeCbl) for MS and adenosyl-Cbl (AdoCbl) for MUT, located in the cytosol and mitochondria, respectively, requires a complex pathway of intracellular processing and trafficking. One of the processing proteins, MMAA (methylmalonic aciduria type A), is implicated in the mitochondrial assembly of AdoCbl into MUT and is defective in children from the cblA complementation group of cobalamin disorders. To characterize the functional interplay between MMAA and MUT, we have crystallized human MMAA in the GDP-bound form and human MUT in the apo, holo, and substrate-bound ternary forms. Structures of both proteins reveal highly conserved domain architecture and catalytic machinery for ligand binding, yet they show substantially different dimeric assembly and interaction, compared with their bacterial counterparts. We show that MMAA exhibits GTPase activity that is modulated by MUT and that the two proteins interact in vitro and in vivo. Formation of a stable MMAA-MUT complex is nucleotide-selective for MMAA (GMPPNP over GDP) and apoenzyme-dependent for MUT. The physiological importance of this interaction is highlighted by a recently identified homoallelic patient mutation of MMAA, G188R, which, we show, retains basal GTPase activity but has abrogated interaction. Together, our data point to a gatekeeping role for MMAA by favoring complex formation with MUT apoenzyme for AdoCbl assembly and releasing the AdoCbl-loaded holoenzyme from the complex, in a GTP-dependent manner.


Neonatal Screening for Congenital Hearing Loss in the North of Jordan; Findings and Implications.
Congenital hearing loss is one of the important illnesses that affect newborns. Early diagnosis and treatment are a challenge for medical authorities in developing countries to improve children's functional, intellectual, emotional, and social abilities. We aimed to study the prevalence of congenital hearing loss in northern Jordan community and identify factors that could affect hearing screening protocol. Prospective cross-sectional study of 1595 infants born in our hospital underwent hearing screening tests. Totally, 104 were tested in NICU and the rest examined in the nursery room using Otoacoustic emission (OAE) test as a primary testing tool. The patients were followed in the three hearing screening phases. Factors affecting screening results were studied and analyzed. The total number of newborns who didn't pass the first OAE test in one or both ears were 90 (5.6%); 69 from the nursery group and 21 from the NICU group. In the 2nd screening phase 21 (23.3%) didn't attend the appointment. Sixty-four passed the second screening OAE test. Five newborns (5.6%) had a second refer result in one or both ears and referred for a diagnostic ABR test. Three infants passed the test and two found to have bilateral hearing loss. Hearing screening test is conducted via a 3-phases-protocol. OAE is used in the first two phases and ABR in the third phase. Hearing results is significantly affected for infants admitted to NICU. The following factors increase OAE fail response: mechanical ventilation for more than 5 days, Hyperbilirubinemia, associated congenital anomalies. Mode of delivery doesn't have statistical significance on hearing screening results.


α7 nicotinic acetylcholine receptor signaling modulates the inflammatory phenotype of fetal brain microglia: first evidence of interference by iron homeostasis.
Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. We hypothesized that agonism on α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia will augment their neuroprotective transcriptome profile, while the antagonistic stimulation of α7nAChR will achieve the opposite. Using an in vivo - in vitro model of developmental programming of neuroinflammation induced by lipopolysaccharide (LPS), we validated this hypothesis in primary fetal sheep microglia cultures re-exposed to LPS in presence of a selective α7nAChR agonist or antagonist. Our RNAseq and protein level findings show that a pro-inflammatory microglial phenotype acquired in vitro by LPS stimulation is reversed with α7nAChR agonistic stimulation. Conversely, antagonistic α7nAChR stimulation potentiates the pro-inflammatory microglial phenotype. Surprisingly, under conditions of LPS double-hit an interference of a postulated α7nAChR - ferroportin signaling pathway may impede this mechanism. These results suggest a therapeutic potential of α7nAChR agonists in early re-programming of microglia in neonates exposed to in utero inflammation via an endogenous cerebral cholinergic anti-inflammatory pathway. Future studies will assess the role of interactions between inflammation-triggered microglial iron sequestering and α7nAChR signaling in neurodevelopment.


Genetics of sporadic amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized clinically by rapidly progressive paralysis leading ultimately to death from respiratory failure. There is substantial evidence suggesting that ALS is a heritable disease, and a number of genes have been identified as being causative in familial ALS. In contrast, the genetics of the much commoner sporadic form of the disease is poorly understood and no single gene has been definitively shown to increase the risk of developing ALS. In this review, we discuss the genetic evidence for each candidate gene that has been putatively associated with increased risk of sporadic ALS. We also review whole genome association studies of ALS and discuss the potential of this methodology for identifying genes relevant to motor neuron degeneration.


Dimensionality and reliability of the self-care of heart failure index scales: further evidence from confirmatory factor analysis.
The Self-Care of Heart Failure Index (SCHFI) is used widely, but issues with reliability have been evident. Cronbach alpha coefficient is usually used to assess reliability, but this approach assumes a unidimensional scale. The purpose of this article is to address the dimensionality and internal consistency reliability of the SCHFI. This was a secondary analysis of data from 629 adults with heart failure enrolled in three separate studies conducted in the northeastern and northwestern United States. Following testing for scale dimensionality using confirmatory factor analysis, reliability was tested using coefficient alpha and alternative options. Confirmatory factor analysis demonstrated that: (a) the Self-Care Maintenance Scale has a multidimensional four-factor structure; (b) the Self-Care Management Scale has a two-factor structure, but the primary factors loaded on a common higher-order factor; and (c) the Self-Care Confidence Scale is unidimensional. Reliability estimates for the three scales, obtained with methods compatible with each scale's dimensionality, were adequate or high. The results of the analysis demonstrate that issues of dimensionality and reliability cannot be separated. Appropriate estimates of reliability that are consistent with the dimensionality of the scale must be used. In the case of the SCHFI, coefficient alpha should not be used to assess reliability of the self-care maintenance and the self-care management scales, due to their multidimensionality. When performing psychometric evaluations, we recommend testing dimensionality before assessing reliability, as well using multiple indices of reliability, such as model-based internal consistency, composite reliability, and omega and maximal reliability coefficients.


Molecular characterization and expression pattern of zona pellucida proteins in gilthead seabream (Sparus aurata).
The developing oocyte is surrounded by an acellular envelope that is composed of 2-4 isoforms of zona pellucida (ZP) proteins. The ZP proteins comprise the ZP1, ZP2, ZP3, and ZPX isoforms. While ZP1 (ZPB) and ZP3 (ZPC) are present in all species, ZP2 (ZPA) is not found in teleost fish and ZPX is not found in mammals. In the present study, we identify and characterize the ZP1, ZP3 and ZPX isoforms of gilthead seabream. Furthermore, by analyzing the conserved domains, which include the external hydrophobic patch and the internal hydrophobic patch, we show that ZP2 and ZPX are closely related isoforms. ZP proteins are synthesized in either the liver or ovary of most teleosts. Only in rainbow trout has it been shown that zp3 has dual transcription sites. In gilthead seabream, all four mRNA isoforms are transcribed in both the liver and ovary, with zp1a, zp1b, and zp3 being highly expressed in the liver, and zpx being primarily expressed in the ovary. However, determination of the ZP proteins in plasma showed high levels of ZP1b, ZP3, and ZPX, with low or non-detectable levels of ZP1a. In similarity to other teleost ZPs, the hepatic transcription of all four ZP isoforms is under estrogenic control. Previously, we have shown that cortisol can potentiate estrogen-induced ZP synthesis in salmonids, and now we show that this is not the case in the gilthead seabream. The present study shows for the first time the endocrine regulation of a teleost ZPX isoform, and demonstrates the dual-organ transcriptional activities of all the ZP proteins in one species.


Heroin abuse accelerates biological aging: a novel insight from telomerase and brain imaging interaction.
Heroin abuse and natural aging exert common influences on immunological cell functioning. This observation led to a recent and untested idea that aging may be accelerated in abusers of heroin. We examined this claim by testing whether heroin use is associated with premature aging at both cellular and brain system levels. A group of abstinent heroin users (n=33) and matched healthy controls (n=30) were recruited and measured on various biological indicators of aging. These measures included peripheral blood telomerase activity, which reflects cellular aging, and both structural and functional measures of brain magnetic resonance imaging. We found that heroin users were characterized by significantly low telomerase activity (0.21 vs 1.78; 88% reduction; t(61)=6.96, P<0.001; 95% confidence interval=1.12-2.02), which interacted with heroin use to affect the structural integrity of gray and white matter of the prefrontal cortex (PFC; AlphaSim corrected P<0.05), a key brain region implicated in aging. Using the PFC location identified from the structural analyses as a 'seed' region, it was further revealed that telomerase activity interacted with heroin use to impact age-sensitive brain functional networks (AlphaSim corrected P<0.05), which correlated with behavioral performance on executive functioning, memory and attentional control (Pearson correlation, all P<0.05). To our knowledge, this study is the first to attempt a direct integration of peripheral molecular, brain system and behavioral measures in the context of substance abuse. The present finding that heroin abuse is associated with accelerated aging at both cellular and brain system levels is novel and forms a unique contribution to our knowledge in how the biological processes of drug abusers may be disrupted.


Local level inequalities in the use of hospital-based maternal delivery in rural South Africa.
There is global concern with geographical and socio-economic inequalities in access to and use of maternal delivery services. Little is known, however, on how local-level socio-economic inequalities are related to the uptake of needed maternal health care. We conducted a study of relative socio-economic inequalities in use of hospital-based maternal delivery services within two rural sub-districts of South Africa. We used both population-based surveillance and facility-based clinical record data to examine differences in the relative distribution of socio-economic status (SES), using a household assets index to measure wealth, among those needing maternal delivery services and those using them in the Bushbuckridge sub-district, Mpumalanga, and Hlabisa sub-district, Kwa-Zulu Natal. We compared the SES distributions in households with a birth in the previous year with the household SES distributions of representative samples of women who had delivered in hospitals in these two sub-districts. In both sub-districts, women in the lowest SES quintile were significantly under-represented in the hospital user population, relative to need for delivery services (8% in user population vs 21% in population in need; p < 0.001 in each sub-district). Exit interviews provided additional evidence on potential barriers to access, in particular the affordability constraints associated with hospital delivery. The findings highlight the need for alternative strategies to make maternal delivery services accessible to the poorest women within overall poor communities and, in doing so, decrease socioeconomic inequalities in utilisation of maternal delivery services.


Exercise-induced bronchoconstriction: diagnosis and management.
Exercise-induced bronchoconstriction describes the narrowing of the airway that occurs with exercise. More than 10 percent of the general population and up to 90 percent of persons previously diagnosed with asthma have exercise-induced bronchoconstriction. Common symptoms include coughing, wheezing, and chest tightness with exercise; however, many athletes will present with nonspecific symptoms, such as fatigue and impaired performance. Spirometry should be performed initially to evaluate for underlying chronic asthma, although results are often normal. An empiric trial of short-acting beta₂ agonists or additional bronchial provocation testing may be necessary to confirm the diagnosis. Nonpharmacologic treatment options include avoiding known triggers, choosing sports with low minute ventilation, warming up before exercising, and wearing a heat exchange mask in cold weather. Short-acting beta₂ agonists are recommended first-line agents for pharmacologic treatment, although leukotriene receptor antagonists or inhaled corticosteroids with or without long-acting beta₂ agonists may be needed in refractory cases. If symptoms persist despite treatment, alternative diagnoses such as cardiac or other pulmonary etiologies, vocal cord dysfunction, or anxiety should be considered.


Experiences of an internet-based support and coaching model for adolescents and young adults with ADHD and autism spectrum disorder -a qualitative study.
There is a great demand for non-medical treatment and support targeting the needs of adolescents and young adults with autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD). There is also a lack of qualitative studies providing in-depth insight into these individuals' own experiences within this area. The current study aimed to explore how adolescents and young adults with ADHD, ASD or both experienced taking part in an internet-based support and coaching intervention. Sixteen participants with ASD, ADHD or both who had participated in an 8-week internet-based support and coaching model, were interviewed using semi-structured interviews. Data was analyzed using qualitative content analysis. Analysis yielded three themes; Deciding to participate, Taking part in the coaching process and The significance of format. Various motives for joining were expressed by participants, such as viewing the technology as familiar and appealing and expecting it to be better suited to their situation. There was also a previously unfulfilled need for support among participants. In deciding to take part in the intervention the coaches' competence and knowledge were considered essential, often in the light of previously negative experiences. Taking part in the coaching process meant feeling reassured by having someone to turn to in view of shared obstacles to seeking and receiving help. The support was used for talking through and receiving advice on matters related to their diagnosis. Findings further revealed appreciation for aspects relating to the format such as communicating through the written word, being in one's own home and an experience of immediacy. Some disadvantages were voiced including incomplete personal interaction and failing technology. There were also suggestions for greater flexibility. The in-depth qualitative data obtained from this study suggest that the current model of support and the internet-based format have specific qualities that could play an important role in the support of adolescents and young adults with ADHD and ASD. Although not a replacement for face-to-face interaction, it could be a promising complement or alternative to other support and treatment options. "Internet-based Support for Young People with ADHD and Autism - a Controlled Study" retrospectively registered in www.clinicaltrials.gov ( ClinicalTrials.gov Identifier: NCT02300597 ) at 2014-11-10.


Analysis of plant leaf metabolites reveals no common response to insect herbivory by Pieris rapae in three related host-plant species.
Studying the biochemical responses of different plant species to insect herbivory may help improve our understanding of the evolution of defensive metabolites found in host plants and their role in plant-herbivore interactions. Untargeted metabolic fingerprints measured as individual mass features were used to compare metabolite reactions in three Brassicales host-plant species (Cleome spinosa, Brassica oleracea, and Lunaria annua) to larval herbivore attack (Pieris rapae; Lepidoptera). Principal component analyses of metabolic fingerprints were able to distinguish among the three plant species and between uneaten control plants and plants that had been eaten. A large number of mass features (1186, 13% of mass features measured in control plants) were common to the three plant species. However, there were few similarities in the mass features that were induced (i.e. changed in abundance) following herbivory. Of the 87 and 68 induced mass features in B. oleracea and C. spinosa, respectively, there were only three that were induced in both plant species. By contrast, L. annua only had one mass feature induced by herbivory, and this was not induced in the other two plant species. The growth of the P. rapae larvae was poorer on the host plant L. annua than on B. oleracea and C. spinosa. The absence of common metabolites among the plants meant these induced responses could not be related to the performance of the herbivore. Thus, the response to herbivory by the same herbivore in these three host plants has evolved to be idiosyncratic in terms of the specific metabolites induced.


Automated bone segmentation from dental CBCT images using patch-based sparse representation and convex optimization.
Cone-beam computed tomography (CBCT) is an increasingly utilized imaging modality for the diagnosis and treatment planning of the patients with craniomaxillofacial (CMF) deformities. Accurate segmentation of CBCT image is an essential step to generate three-dimensional (3D) models for the diagnosis and treatment planning of the patients with CMF deformities. However, due to the poor image quality, including very low signal-to-noise ratio and the widespread image artifacts such as noise, beam hardening, and inhomogeneity, it is challenging to segment the CBCT images. In this paper, the authors present a new automatic segmentation method to address these problems. To segment CBCT images, the authors propose a new method for fully automated CBCT segmentation by using patch-based sparse representation to (1) segment bony structures from the soft tissues and (2) further separate the mandible from the maxilla. Specifically, a region-specific registration strategy is first proposed to warp all the atlases to the current testing subject and then a sparse-based label propagation strategy is employed to estimate a patient-specific atlas from all aligned atlases. Finally, the patient-specific atlas is integrated into a maximum a posteriori probability-based convex segmentation framework for accurate segmentation. The proposed method has been evaluated on a dataset with 15 CBCT images. The effectiveness of the proposed region-specific registration strategy and patient-specific atlas has been validated by comparing with the traditional registration strategy and population-based atlas. The experimental results show that the proposed method achieves the best segmentation accuracy by comparison with other state-of-the-art segmentation methods. The authors have proposed a new CBCT segmentation method by using patch-based sparse representation and convex optimization, which can achieve considerably accurate segmentation results in CBCT segmentation based on 15 patients.


Alterations in contractile properties of tongue muscles in old rats.
Fatigue and weakness are well-known signs of aging that are related to sarcopenia, or loss of skeletal muscle mass, organization, and strength. Sarcopenia may affect swallowing. The tongue plays a vital role in swallowing, but there is limited knowledge regarding age-related changes in lingual muscle contractile properties. Our purpose was to determine whether alterations in tongue force, temporal features of tongue muscle contraction, and fatigability are manifested as a function of aging in old rats. We evaluated tongue muscle contractile properties in young and old Fischer 344/Brown Norway rats. Contractions were elicited via bilateral electrical stimulation of the hypoglossal nerves. Maximum tongue forces and fatigability were not significantly altered in old animals, but aging was associated with significantly longer twitch contraction time and longer half-decay recovery time intervals (p < .01). The results indicated that old animals generated sufficient maximum tongue forces, but were slower in achieving these forces than young animals. These findings are consistent with reports of altered temporal parameters of tongue actions during swallowing in humans, and suggest that a disruption in the timing of muscle contraction onset and recovery may contribute to the altered tongue kinetics observed with aging.


Emergency bedside ultrasound-benefits as well as caution: Part 2: Echocardiography.
Critical care echocardiography (CCE) has become an important component of general critical care ultrasonography, and a current review of its performance is presented. Basic CCE should be performed as a goal-directed examination to better identify specific signs and to answer important clinical questions concerning acute hemodynamic concerns. It has evolved in the ICU and also in the emergency department not only for improved diagnostic capability but also as an effective part of the triage process. It remains an efficacious procedure even in patients with respiratory failure when combined with lung ultrasonography. Numerous acronyms were proposed, but in all cases, CCE responds to the same rules as fundamental echocardiography. Basic CCE requires accessible and comprehensive training for physicians and is mandatory for all intensivists. Development of pocket echo devices may increase the use of basic CCE as has miniaturization of other medical technologies. Performance should be managed by guidelines, and the CCE training program should be standardized worldwide. More trials are welcome to evaluate its impact on patient outcomes. Thanks to its ability to quickly obtain a diagnostic orientation at the bedside and to implement targeted therapy, basic CCE over the past decade has become an essential tool for hemodynamic assessment of the cardiopulmonary unstable patient. Its more recent incorporation into the education of trainees in medical school and residencies/fellowships has reinforced its perceived importance in critical care management, despite the relative paucity as yet of rigorous scientific evidence demonstrating positive outcome modification from its use.


Barriers and enablers to detection and management of chronic kidney disease in primary healthcare: a systematic review.
Chronic kidney disease (CKD) is growing population health concern worldwide, and with early identification and effective management, kidney disease progression can be slowed or prevented. Most patients with risk factors for chronic kidney disease are treated within primary healthcare. Therefore, it is important to understand how best to support primary care providers (PC-P) to detect and manage chronic kidney disease. The aim of this systematic review was to evaluate barriers and enablers to the diagnosis and management of CKD in primary care. A systematic review of qualitative research on the barriers and/or enablers to detection and/or management of CKD in adults within primary healthcare was conducted. The databases Medline (EBSCO), PubMed, Cochrane CENTRAL, CINAHL (EBSCO) and Joanna Briggs Institute Evidence Based Practice (Ovid) were searched until 27th August 2019. Barriers and/or enablers reported in each study were identified, classified into themes, and categorised according to the Theoretical Domains Framework. A total of 20 studies were included in this review. The most commonly reported barriers related to detection and management of CKD in primary care were categorised into the 'Environmental context and resources' domain (n = 16 studies). Overall, the most common barrier identified was a lack of time (n = 13 studies), followed by a fear of delivering a diagnosis of CKD, and dissatisfaction with CKD guidelines (both n = 10 studies). Overall, the most common enabler identified was the presence of supportive technology to identify and manage CKD (n = 7 studies), followed by the presence of a collaborative relationship between members of the healthcare team (n = 5 studies). This systematic review identified a number of barriers and enablers which PC-P face when identifying and managing CKD. The findings of this review suggest a need for time-efficient strategies that promote collaboration between members of the healthcare team, and practice guidelines which consider the frequently co-morbid nature of CKD. Enhanced collaboration between PC-P and nephrology services may also support PC-Ps when diagnosing CKD in primary care, and facilitate improved patient self-management.


The lipocalin protein family: structure and function.
The lipocalin protein family is a large group of small extracellular proteins. The family demonstrates great diversity at the sequence level; however, most lipocalins share three characteristic conserved sequence motifs, the kernel lipocalins, while a group of more divergent family members, the outlier lipocalins, share only one. Belying this sequence dissimilarity, lipocalin crystal structures are highly conserved and comprise a single eight-stranded continuously hydrogen-bonded antiparallel beta-barrel, which encloses an internal ligand-binding site. Together with two other families of ligand-binding proteins, the fatty-acid-binding proteins (FABPs) and the avidins, the lipocalins form part of an overall structural superfamily: the calycins. Members of the lipocalin family are characterized by several common molecular-recognition properties: the ability to bind a range of small hydrophobic molecules, binding to specific cell-surface receptors and the formation of complexes with soluble macromolecules. The varied biological functions of the lipocalins are mediated by one or more of these properties. In the past, the lipocalins have been classified as transport proteins; however, it is now clear that the lipocalins exhibit great functional diversity, with roles in retinol transport, invertebrate cryptic coloration, olfaction and pheromone transport, and prostaglandin synthesis. The lipocalins have also been implicated in the regulation of cell homoeostasis and the modulation of the immune response, and, as carrier proteins, to act in the general clearance of endogenous and exogenous compounds.


Changes in chronic pain severity over time: the Chronic Pain Grade as a valid measure.
Our understanding of the natural history of chronic pain in the community is limited. This is partly due to the lack of a validated measure of chronic pain severity known to be responsive to change over time. The Chronic Pain Grade questionnaire has been shown to be valid and reliable for use in a general population as a self-completion questionnaire. However, its reliability and validity for use in longitudinal studies and its responsiveness to change over time has not yet been assessed. We undertook a postal survey designed to test the responsiveness and the validity of the Chronic Pain Grade questionnaire over time. A random sample of 560 chronic pain patients, aged 25 years and over was drawn from an existing cohort and stratified for age, gender and chronic pain severity. Subjects were re-surveyed by a postal self-completion questionnaire consisting of the Chronic Pain Grade and the SF-36 general health questionnaire, which is known to be responsive to change in health over time. To test whether changes in CPG scores correlated with changes in SF-36 scores, Spearman's rank correlation coefficients were calculated. A response rate of 86% was achieved for the follow-up study. The majority of SF-36 scores changed in the hypothesized directions. Changes in CPG scores were significantly correlated with changes in most of the SF-36 domains. We concluded that the CPG is a useful and valid objective instrument for measuring change in severity of chronic pain over time and could be used in longitudinal studies of chronic pain severity.


Recommendations to Improve Employee Thermal Comfort When Working in 40°F Refrigerated Cold Rooms.
Cold rooms are commonly used for food storage and preparation, and are usually kept around 40°F following food safety guidelines. Some food preparation employees may spend 8 or more hours inside cold rooms. These employees may not be aware of the risks associated with mildly cold temperatures, dampness, and limited ventilation. We performed an evaluation of cold rooms at an airline catering facility because of concerns with exposure to cold temperatures. We spoke with and observed employees in two cold rooms, reviewed daily temperature logs, evaluated employee's physical activity, work/rest schedule, and protective clothing. We measured temperature, percent relative humidity, and air velocities at different work stations inside the cold rooms. We concluded that thermal comfort concerns perceived by cold room employees may have been the result of air drafts at their workstations, insufficient use of personal protective equipment due to dexterity concerns, work practices, and lack of knowledge about good health and safety practices in cold rooms. These moderately cold work conditions with low air velocities are not well covered in current occupational health and safety guidelines, and wind chill calculations do not apply. We provide practical recommendations to improve thermal comfort of cold room employees. Engineering control recommendations include the redesigning of air deflectors and installing of suspended baffles. Administrative controls include the changing out of wet clothing, providing hand warmers outside of cold rooms, and educating employees on cold stress. We also recommended providing more options on personal protective equipment. However, there is a need for guidelines and educational materials tailored to employees in moderately cold environments to improve thermal comfort and minimize health and safety problems.


Immunodiagnosis of Ehrlichia canis infection with recombinant proteins.
Ehrlichia canis causes a potentially fatal rickettsial disease of dogs that requires rapid and accurate diagnosis in order to initiate appropriate therapy leading to a favorable prognosis. We recently reported the cloning of two immunoreactive E. canis proteins, P28 and P140, that were applicable for serodiagnosis of the disease. In the present study we cloned a new immunoreactive E. canis surface protein gene of 1,170 bp, which encodes a protein with a predicted molecular mass of 42.6 kDa (P43). The P43 gene was not detected in E. chaffeensis DNA by Southern blot, and antisera against recombinant P43 (rP43) did not react with E. chaffeensis as detected by indirect fluorescent antibody (IFA) assay. Forty-two dogs exhibiting signs and/or hematologic abnormalities associated with canine ehrlichiosis were tested by IFA assay and by recombinant Western immunoblot. Among the 22 samples that were IFA positive for E. canis, 100% reacted with rP43, 96% reacted with rP28, and 96% reacted with rP140. The specificity of the recombinant proteins compared to the IFAs was 96% for rP28, 88% for P43 and 63% for P140. The results of this study demonstrate that the rP43 and rP28 are sensitive and reliable serodiagnostic antigens for E. canis infections.


Working well with others: the prediction of students' clinical performance.
The purpose of this study was to determine whether multivariate measurement at the initiation of students' training in occupational therapy could predict clinical performance in Level II fieldwork. Forty-five students in their first semester were videotaped in pairs and completed personality and emotional communication measures related to the potential for working well with others. Two years later, the students' clinical performance was rated by Level II fieldwork supervisors. In the physical rehabilitation fieldwork setting, students who were conventional and quietly concerned were rated higher than students who were unconventional, talkative, and less other-centered. In the pediatric rehabilitation setting, students who were autonomous and nonverbally perceptive were rated higher than students who were interdependent and less perceptive. In the psychiatric rehabilitation setting, students who were emotionally reactive to facial but not body cues of emotion were rated higher than students who had low emotional reactivity and were less perceptive to facial but not body cues. The varying institutional cultures, types of patients, and roles of occupational therapy across fieldwork settings may require different sets of attributes for working well with others. Educators and supervisors should be aware of these possible differences when fashioning Level I fieldwork experiences.


ntPET: a new application of PET imaging for characterizing the kinetics of endogenous neurotransmitter release.
We present a new application of positron emission tomography ("ntPET" or "neurotransmitter PET") designed to recover temporal patterns of neurotransmitter release from dynamic data. Our approach employs an enhanced tracer kinetic model that describes uptake of a labeled dopamine D2/D3 receptor ligand in the presence of a time-varying rise and fall in endogenous dopamine. Data must be acquired during both baseline and stimulus (transient dopamine release) conditions. Data from a reference region in both conditions are used as an input function, which alleviates the need for any arterial blood sampling. We use simulation studies to demonstrate the ability of the method to recover the temporal characteristics of an increase in dopamine concentration that might be expected following a drug treatment. The accuracy and precision of the method-as well as its potential for false-positive responses due to noise or changes in blood flow-were examined. Finally, we applied the ntPET method to small-animal imaging data in order to produce the first noninvasive assay of the time-varying release of dopamine in the rat striatum following alcohol.


Magnetic resonance imaging studies in early-onset bipolar disorder: a critical review.
Neuroimaging studies of early-onset bipolar disorder (BD) are important in order to establish a fuller understanding of the underlying pathophysiology of the illness. The advantages of studying BD in children and adolescents include the relative absence of some confounds present in adult-onset research, such as lengthy duration of illness and exposure to treatments, greater number of mood episodes, and the presence of substance abuse or dependence. Finally, studying youths with the disorder may enhance our knowledge about the neural mechanisms of affective dysregulation and may specifically elucidate whether there are abnormalities that are unique to the early-onset form of the illness. PubMed was used to identify peer-reviewed publications from the past 15 years (January 1990 to January 2005) that used brain-imaging techniques (anatomic, functional, and biochemical) to research early-onset BD. Eleven studies using anatomic magnetic resonance imaging (MRI), seven using magnetic resonance spectroscopy (MRS), and two using functional MRI (fMRI) were identified. Structural abnormalities were reported in total cerebral, white matter, superior temporal gyrus, putamen, thalamus, amygdala, and hippocampal volumes. Deficits in cortical gray matter were also reported. Using MRS, abnormalities were reported in the dorsolateral prefrontal cortex, anterior cingulate, and basal ganglia. One fMRI study found increased activation in the putamen and thalamus of BD youths compared to controls, and a second found abnormal prefrontal-subcortical activation in familial pediatric BD. Published MRI studies of early-onset BD are few. Nonetheless, extant data implicate abnormalities in brain regions thought to regulate mood and cognition. Synthesis of the findings into an overall model of anatomic and functional disruption is difficult due to the methodological variations among studies and the limitations of individual studies, such as the use of small sample sizes, the heterogeneity of sample characteristics, and the wide range of brain structures selected for analysis. Recommendations are offered to guide future research. It will be important for future studies to reproduce prior findings and determine which findings are unique to early-onset BD, relative to adult-onset illness. In addition, studies will need to establish the extent to which early-onset BD may overlap with comorbid disruptive, mood, anxiety, or psychotic disorders.


How actin binds and assembles onto plasma membranes from Dictyostelium discoideum.
We have shown previously (Schwartz, M. A., and E. J. Luna. 1986. J. Cell Biol. 102: 2067-2075) that actin binds with positive cooperativity to plasma membranes from Dictyostelium discoideum. Actin is polymerized at the membrane surface even at concentrations well below the critical concentration for polymerization in solution. Low salt buffer that blocks actin polymerization in solution also prevents actin binding to membranes. To further explore the relationship between actin polymerization and binding to membranes, we prepared four chemically modified actins that appear to be incapable of polymerizing in solution. Three of these derivatives also lost their ability to bind to membranes. The fourth derivative (EF actin), in which histidine-40 is labeled with ethoxyformic anhydride, binds to membranes with reduced affinity. Binding curves exhibit positive cooperativity, and cross-linking experiments show that membrane-bound actin is multimeric. Thus, binding and polymerization are tightly coupled, and the ability of these membranes to polymerize actin is dramatically demonstrated. EF actin coassembles weakly with untreated actin in solution, but coassembles well on membranes. Binding by untreated actin and EF actin are mutually competitive, indicating that they bind to the same membrane sites. Hill plots indicate that an actin trimer is the minimum assembly state required for tight binding to membranes. The best explanation for our data is a model in which actin oligomers assemble by binding to clustered membrane sites with successive monomers on one side of the actin filament bound to the membrane. Individual binding affinities are expected to be low, but the overall actin-membrane avidity is high, due to multivalency. Our results imply that extracellular factors that cluster membrane proteins may create sites for the formation of actin nuclei and thus trigger actin polymerization in the cell.


Model-driven evaluation of the production potential for growth-coupled products of Escherichia coli.
Integrated approaches utilizing in silico analyses will be necessary to successfully advance the field of metabolic engineering. Here, we present an integrated approach through a systematic model-driven evaluation of the production potential for the bacterial production organism Escherichia coli to produce multiple native products from different representative feedstocks through coupling metabolite production to growth rate. Designs were examined for 11 unique central metabolism and amino acid targets from three different substrates under aerobic and anaerobic conditions. Optimal strain designs were reported for designs which possess maximum yield, substrate-specific productivity, and strength of growth-coupling for up to 10 reaction eliminations (knockouts). In total, growth-coupled designs could be identified for 36 out of the total 54 conditions tested, corresponding to eight out of the 11 targets. There were 17 different substrate/target pairs for which over 80% of the theoretical maximum potential could be achieved. The developed method introduces a new concept of objective function tilting for strain design. This study provides specific metabolic interventions (strain designs) for production strains that can be experimentally implemented, characterizes the potential for E. coli to produce native compounds, and outlines a strain design pipeline that can be utilized to design production strains for additional organisms.


Fructose-induced mTORC1 Activation Promotes Pancreatic Cancer Progression through Inhibition of Autophagy.
Excessive fructose intake is associated with the occurrence, progression, and poor prognosis of various tumors. A better understanding of the mechanisms underlying the functions of fructose in cancer could facilitate the development of better treatment and prevention strategies. In this study, we investigated the functional association between fructose utilization and pancreatic ductal adenocarcinoma (PDAC) progression. Fructose could be taken up and metabolized by PDAC cells and provided an adaptive survival mechanism for PDAC cells under glucose deficient conditions. GLUT5-mediated fructose metabolism maintained the survival, proliferation, and invasion capacities of PDAC cells in vivo and in vitro. Fructose metabolism not only provided ATP and biomass to PDAC cells but also conferred metabolic plasticity to the cells, making them more adaptable to the tumor microenvironment. Mechanistically, fructose activated AMPK-mTORC1 signaling pathway to inhibit glucose deficiency-induced autophagic cell death. Moreover, the fructose specific transporter GLUT5 was highly expressed in PDAC tissues and was an independent marker of disease progression in PDAC patients. These findings provide mechanistic insights into the role of fructose in promoting PDAC progression and offer potential strategies for targeting metabolism to treat PDAC.


High Versatility and Stability of Mechanochemically Synthesized Halide Perovskite Powders for Optoelectronic Devices.
We show that mechanochemically synthesized halide perovskite powders from a ball milling approach can be employed to fabricate a variety of lead halide perovskites with exceptional intrinsic stability. Our MAPbI3 powder exhibits higher thermal stability than conventionally processed thin films, without degradation after more than two and a half years of storage and only negligible degradation after heat treatment at 220 °C for 14 h. We further show facile recovery strategies of nonphase-pure powders by simple remilling or mild heat treatment. Moreover, we demonstrate the mechanochemical synthesis of phase-pure mixed perovskite powders, such as (Cs0.05FA0.95PbI3)0.85(MAPbBr3)0.15, from either the individual metal and organic halides or from readily prepared ternary perovskites, regardless of the precursor phase purity. Adding potassium iodide (KI) to the milling process successfully passivated the powders. We also succeeded in preparing a precursor solution on the basis of the powders and obtained uniform thin films for integration into efficient perovskite solar cells from spin-coating this solution. We find the KI passivation remains in the devices, leading to improved performance and significantly reduced hysteresis. Our work thus demonstrates the potential of mechanochemically synthesized halide perovskite powders for long-time storage and upscaling, further paving the way toward commercialization of perovskite-based optoelectronic devices.


Mechanical influence of static versus dynamic loadings on parametrical analysis of plasticized ethyl cellulose films.
The object of this study was to investigate the influence of static and dynamic forces on mechanical properties of the biocompatible polymer ethyl cellulose. Similar polymeric films containing 40% (w/w) of the plasticizer dibutyl sebacate were subjected to tensile forces at different velocities. The average Young's modulus and the variation of yield strength, strain, and strain energy at different velocities complied with the pre-established theories of dynamic loadings. The ultimate strength and the yield strength and/or strain displayed linearity with the velocity, though the ultimate strain and therefore, the plastic and/or ultimate working energies proved non-linear pseudo-Michaelis-Menten behavior. The speculation was that achieving the maximum displacement would probably be the most important cause of failure. Finally the most suitable velocity at which the data would obtain the most demonstrable stress-strain diagrams was selected: tensile forces at almost low velocities, best illustrated as static forces, proved immature failure of the specimens during or shortly after the yield; so that the specimen resembled as a brittle material. High velocities of loadings were also avoided since the strain would usually approach the plateau and would therefore disrupt the rational correlation between forces and displacements during the end region of the curve.


The prevalence and risk factors of dysmenorrhea.
Dysmenorrhea is a common menstrual complaint with a major impact on women's quality of life, work productivity, and health-care utilization. A comprehensive review was performed on longitudinal or case-control or cross-sectional studies with large community-based samples to accurately determine the prevalence and/or incidence and risk factors of dysmenorrhea. Fifteen primary studies, published between 2002 and 2011, met the inclusion criteria. The prevalence of dysmenorrhea varies between 16% and 91% in women of reproductive age, with severe pain in 2%-29% of the women studied. Women's age, parity, and use of oral contraceptives were inversely associated with dysmenorrhea, and high stress increased the risk of dysmenorrhea. The effect sizes were generally modest to moderate, with odds ratios varying between 1 and 4. Family history of dysmenorrhea strongly increased its risk, with odds ratios between 3.8 and 20.7. Inconclusive evidence was found for modifiable factors such as cigarette smoking, diet, obesity, depression, and abuse. Dysmenorrhea is a significant symptom for a large proportion of women of reproductive age; however, severe pain limiting daily activities is less common. This review confirms that dysmenorrhea improves with increased age, parity, and use of oral contraceptives and is positively associated with stress and family history of dysmenorrhea.


Beyond 9+0: noncanonical axoneme structures characterize sensory cilia from protists to humans.
The intracellular amastigote stages of parasites such as Leishmania are often referred to as aflagellate. They do, however, possess a short axoneme of cryptic function. Here, our examination of the structure of this axoneme leads to a testable hypothesis of its role in the cell biology of pathogenicity. We show a striking similarity between the microtubule axoneme structure of the Leishmania mexicana parasite infecting a macrophage and vertebrate primary cilia. In both, the 9-fold microtubule doublet symmetry is broken by the incursion of one or more microtubule doublets into the axoneme core, giving rise to an architecture that we term here the 9v (variable) axoneme. Three-dimensional reconstructions revealed that no particular doublet initiated the symmetry break, and moreover it often involved 2 doublets. The tip of the L. mexicana flagellum was frequently intimately associated with the macrophage vacuole membrane. We propose that the main function of the amastigote flagellum is to act as a sensory organelle with important functions in host-parasite interactions and signaling in the intracellular stage of the L. mexicana life cycle.


Nonsurgical splenectomy.
Surgical removal of the spleen is a well established procedure which is indicated for various hematologic disorders. Patients who are poor surgical risks may be considered for nonsurgical "splenectomy* using an embolization technique. This new technique should only be performed in patients who represent high surgical risk. The possible effectiveness of this procedure should be previously tested with the use of intraarterial vasopressin infusion. The following conditions should be fulfilled: (1) small emboli should be used to eliminate collateral flow; (2) the procedure should be carried out in two or three stages to minimize flank pain and the risk of massive splenic necrosis; and (3) most important, the spleen should be surgically removed as soon as the hematologic condition has improved in order to prevent splenic abscess formation. Splenic embolization was carried out in three patients with hypersplenism. One survived but was not helped by the procedure. Two died, one definitely and the other possibly as a result of the embolization. These discouraging results were reproduced in the animal laboratory where 11 of 14 animals developed severe complications, six of which were acutely fatal. The high incidence of late complications precludes non-surgical splenectomy as a routine procedure, but it can be carried out provided the spleen can be removed surgically.


Siglec-H-Deficient Mice Show Enhanced Type I IFN Responses, but Do Not Develop Autoimmunity After Influenza or LCMV Infections.
Siglec-H is a DAP12-associated receptor on plasmacytoid dendritic cells (pDCs) and microglia. Siglec-H inhibits TLR9-induced IFN-α production by pDCs. Previously, it was found that Siglec-H-deficient mice develop a lupus-like severe autoimmune disease after persistent murine cytomegalovirus (mCMV) infection. This was due to enhanced type I interferon responses, including IFN-α. Here we examined, whether other virus infections can also induce autoimmunity in Siglec-H-deficient mice. To this end we infected Siglec-H-deficient mice with influenza virus or with Lymphocytic Choriomeningitis virus (LCMV) clone 13. With both types of viruses we did not observe induction of autoimmune disease in Siglec-H-deficient mice. This can be explained by the fact that both types of viruses are ssRNA viruses that engage TLR7, rather than TLR9. Also, Influenza causes an acute infection that is rapidly cleared and the chronicity of LCMV clone 13 may not be sufficient and may rather suppress pDC functions. Siglec-H inhibited exclusively TLR-9 driven type I interferon responses, but did not affect type II or type III interferon production by pDCs. Siglec-H-deficient pDCs showed impaired Hck expression, which is a Src-family kinase expressed in myeloid cells, and downmodulation of the chemokine receptor CCR9, that has important functions for pDCs. Accordingly, Siglec-H-deficient pDCs showed impaired migration towards the CCR9 ligand CCL25. Furthermore, autoimmune-related genes such as Klk1 and DNase1l3 are downregulated in Siglec-H-deficient pDCs as well. From these findings we conclude that Siglec-H controls TLR-9-dependent, but not TLR-7 dependent inflammatory responses after virus infections and regulates chemokine responsiveness of pDCs.


Amino acid-insensitive mTORC1 regulation enables nutritional stress resilience in hematopoietic stem cells.
The mTOR pathway is a critical determinant of cell persistence and growth wherein mTOR complex 1 (mTORC1) mediates a balance between growth factor stimuli and nutrient availability. Amino acids or glucose facilitates mTORC1 activation by inducing RagA GTPase recruitment of mTORC1 to the lysosomal outer surface, enabling activation of mTOR by the Ras homolog Rheb. Thereby, RagA alters mTORC1-driven growth in times of nutrient abundance or scarcity. Here, we have evaluated differential nutrient-sensing dependence through RagA and mTORC1 in hematopoietic progenitors, which dynamically drive mature cell production, and hematopoietic stem cells (HSC), which provide a quiescent cellular reserve. In nutrient-abundant conditions, RagA-deficient HSC were functionally unimpaired and upregulated mTORC1 via nutrient-insensitive mechanisms. RagA was also dispensable for HSC function under nutritional stress conditions. Similarly, hyperactivation of RagA did not affect HSC function. In contrast, RagA deficiency markedly altered progenitor population function and mature cell output. Therefore, RagA is a molecular mechanism that distinguishes the functional attributes of reactive progenitors from a reserve stem cell pool. The indifference of HSC to nutrient sensing through RagA contributes to their molecular resilience to nutritional stress, a characteristic that is relevant to organismal viability in evolution and in modern HSC transplantation approaches.


The fitness landscape of the African Salmonella Typhimurium ST313 strain D23580 reveals unique properties of the pBT1 plasmid.
We have used a transposon insertion sequencing (TIS) approach to establish the fitness landscape of the African Salmonella enterica serovar Typhimurium ST313 strain D23580, to complement our previous comparative genomic and functional transcriptomic studies. We used a genome-wide transposon library with insertions every 10 nucleotides to identify genes required for survival and growth in vitro and during infection of murine macrophages. The analysis revealed genomic regions important for fitness under two in vitro growth conditions. Overall, 724 coding genes were required for optimal growth in LB medium, and 851 coding genes were required for growth in SPI-2-inducing minimal medium. These findings were consistent with the essentiality analyses of other S. Typhimurium ST19 and S. Typhi strains. The global mutagenesis approach also identified 60 sRNAs and 413 intergenic regions required for growth in at least one in vitro growth condition. By infecting murine macrophages with the transposon library, we identified 68 genes that were required for intra-macrophage replication but did not impact fitness in vitro. None of these genes were unique to S. Typhimurium D23580, consistent with a high conservation of gene function between S. Typhimurium ST313 and ST19 and suggesting that novel virulence factors are not involved in the interaction of strain D23580 with murine macrophages. We discovered that transposon insertions rarely occurred in many pBT1 plasmid-encoded genes (36), compared with genes carried by the pSLT-BT virulence plasmid and other bacterial plasmids. The key essential protein encoded by pBT1 is a cysteinyl-tRNA synthetase, and our enzymological analysis revealed that the plasmid-encoded CysRSpBT1 had a lower ability to charge tRNA than the chromosomally-encoded CysRSchr enzyme. The presence of aminoacyl-tRNA synthetases in plasmids from a range of Gram-negative and Gram-positive bacteria suggests that plasmid-encoded essential genes are more common than had been appreciated.


Does normal pump function belie muscle dysfunction in patients with chronic severe mitral regurgitation?
Left ventricular cineangiography and micromanometry were performed simultaneously in 27 patients with chronic severe mitral regurgitation (MR group) and in 23 normal subjects (NL group). Stress (sigma) and volume (V) were computed frame by frame. Measurements were repeated after pharmacologic load manipulation in subsets of MR (n = 10) and NL (n = 11) groups. An inverse relationship (r = -.90) between EFc (ejection fraction determined from a common preload) and sigma es (afterload) was observed for the NL group. For the MR group, 10 of 14 with EFs less than 0.60 and four of 13 with EFs greater than 0.60 had muscle dysfunction, falling below the 95% prediction band of the normal EFc-sigma es relationship. Maximum myocardial stiffness (maxEN) determined from the end-systolic stress-strain relationship of Mirsky, sigma es = maxEN.gamma.loge(Ves/Vo), was 1398 +/- 716 in the MR (n = 10) vs 1165 +/- 394 in the NL group (n = 11, NS). EF was 0.62 +/- 0.13 in the MR and 0.65 +/- 0.08 in the NL group. Mitral valve surgery was performed on 19 of the patients with MR. All survived and all but one were symptomatically improved; that patient required reoperation in the early postoperative period because of transverse midventricular disruption. Thus, contractile function as assessed by stress-strain and EFc-afterload relationships is frequently normal in individuals with MR when EF is normal. When EF is depressed, contractile function as assessed by EFc-afterload relationships is frequently depressed, but this does not preclude a satisfactory surgical result.


Effect of celecoxib on cyclooxygenase-2 expression and possible variants in a patient with Barrett's esophagus.
Cyclooxygenase-2 (COX-2) expression is increased in metaplastic and dysplastic Barrett's esophageal epithelium and it is thought that selective COX-2 inhibitors could offer hope as chemoprevention therapy. The aim of the study was to investigate the in vivo effect of celecoxib on COX-2 expression in patients with Barrett's esophagus and no recent history of non-steroidal anti-inflammatory drug use. Endoscopic mucosal biopsy specimens were collected at baseline and after 28 days of therapy in a patient treated with celecoxib 200 mg twice daily. Samples were analyzed for COX-2 expression by immunoblot analysis with chemiluminescence detection. COX-2 expression was found to decline 20% and 44% at two different biopsy sites compared to the baseline sample. Longer exposures revealed a number of previously unidentified proteins above and below the 67 kDa COX-2 protein including 38 kDa and 45 kDa proteins which were present only at study completion consistent with up-regulation after celecoxib therapy. Further investigations of the 38 kDa and 45 kDa proteins were undertaken using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with immunoblot and MALDI-TOF (matrix assisted laser desorption ionization - time of flight) analysis but no matches were found and results were inconclusive. Unmatched masses from MALDI-TOF peptide mass fingerprinting were compared with human COX-2 (67 kDa) and COX-2b (39 kDa) using unspecific cleavage. Peptide sequence homology with COX-2 and COX-2b was found for a length of 19 amino acids. Based on immunodetection, molecular weight and equivical MALDI-TOF results, one of these up-regulated proteins may be COX-2b.


Effects of antioxidants on oxidant-induced sister chromatid exchange formation.
Stimulated human phagocytes produce sister chromatid exchanges in cultured mammalian cells by a mechanism involving oxygen metabolites. Experiments were designed to determine whether antioxidants inhibit this process. Superoxide dismutase, catalase, and hydroxyl radical scavengers (benzoate, mannitol) protected target Chinese hamster ovary cells from phagocyte-induced sister chromatid exchanges, implicating the involvement of hydroxyl radicals in this chromosomal damage. N-acetylcysteine and beta-carotene were also protective. alpha-Tocopherol (greater than 5 microM) protected target cells exposed to phagocytes but not to enzymatically generated oxidants when the vitamin was added just before the source of oxygen radicals, suggesting, as reported by others, that the principal action of tocopherol in this setting was to inhibit the release of oxidants from phagocytes. On the other hand, cultivation of target cells with supplemental tocopherol protected them from the toxic effects of the enzymatic oxidant-producing system, indicating a role for membrane-associated free radicals in the mechanism of sister chromatid exchange induction. Low concentrations of sodium selenite (0.1-1.0 microM) protected the target cells. However, higher concentrations (10 microM) of selenite had no effect on oxidant-induced sister chromatid exchange formation, and 0.1 mM selenite increased the number of exchanges. Sodium selenite concentrations of 0.1 mM also decreased the intracellular glutathione concentration of target cells during an oxidant stress, and reducing target cell glutathione concentrations with buthionine sulfoximine increased their sensitivity to oxygen-related chromosomal damage. Therefore, the potentiation of oxygen radical-induced chromosomal damage observed with high concentrations of selenite may result from a decrease in the thiol antioxidant defense systems within the cell. The findings suggest that the hydroxyl radical has an important role in the production of phagocyte-induced cytogenetic injury, membrane-derived intermediates may be involved, depletion of intracellular glutathione renders cells more susceptible to this injury, and supplementation of target cells with antioxidants can protect them from oxygen radical-generated chromosomal injury.


Bipolar disorder. II: personality and age of onset.
The aim of this study was to examine whether personality i.e. temperament and character interacts with age of onset in bipolar disorder. Bipolar patients were recruited among in- and outpatients from lithium dispensaries of northern Sweden. Patients were diagnosed according to DSM-IV criteria for bipolar disorder type I and II. Temperament and Character Inventory (TCI) was used for measuring personality. TCI was administered to 100 lithium treated bipolar patients and 100 controls. Treatment response was significantly lower (p = 0.005) in patients with early onset compared with late onset. Family history (p = 0.013) and suicide attempts (p = 0.001) were also significantly more common in patients with early onset. Further, patients with early onset were significantly higher (p = 0.045) in the temperament factor harm avoidance (HA) than patients with late onset, but the difference was weak. Patients with early onset had more fear of uncertainty (HA2; p = 0.022) and were more shy (HA3; p = 0.030). Bipolar I patients showed similar results as those in the total bipolar group (I and II), with significantly higher HA (p = 0.019, moderate difference), HA2 (p = 0.015) and HA3 (p = 0.043) in patients with early onset compared with late onset. Bipolar II patients showed no differences between early and late age of onset but the groups are small and the results are therefore uncertain. Early age of onset in bipolar disorder was correlated to an increase in severity, family history, poorer treatment response and poorer prognosis. Early onset was also correlated to personality.


The botrydial biosynthetic gene cluster of Botrytis cinerea displays a bipartite genomic structure and is positively regulated by the putative Zn(II)2Cys6 transcription factor BcBot6.
Botrydial (BOT) is a non-host specific phytotoxin produced by the polyphagous phytopathogenic fungus Botrytis cinerea. The genomic region of the BOT biosynthetic gene cluster was investigated and revealed two additional genes named Bcbot6 and Bcbot7. Analysis revealed that the G+C/A+T-equilibrated regions that contain the Bcbot genes alternate with A+T-rich regions made of relics of transposable elements that have undergone repeat-induced point mutations (RIP). Furthermore, BcBot6, a Zn(II)2Cys6 putative transcription factor was identified as a nuclear protein and the major positive regulator of BOT biosynthesis. In addition, the phenotype of the ΔBcbot6 mutant indicated that BcBot6 and therefore BOT are dispensable for the development, pathogenicity and response to abiotic stresses in the B. cinerea strain B05.10. Finally, our data revealed that B. pseudocinerea, that is also polyphagous and lives in sympatry with B. cinerea, lacks the ability to produce BOT. Identification of BcBot6 as the major regulator of BOT synthesis is the first step towards a comprehensive understanding of the complete regulation network of BOT synthesis and of its ecological role in the B. cinerea life cycle.


Characterization of the Microbiome along the Gastrointestinal Tract of Growing Turkeys.
The turkey microbiome is largely understudied, despite its relationship with bird health and growth, and the prevalence of human pathogens such as Campylobacter spp. In this study we investigated the microbiome within the small intestine (SI), caeca (C), large intestine (LI), and cloaca (CL) of turkeys at 6, 10, and 16 weeks of age. Eight turkeys were dissected within each age category and the contents of the SI, C, LI, and CL were harvested. 16S rDNA based QPCR was performed on all samples and samples for the four locations within three birds/age group were sequenced using ion torrent-based sequencing of the 16S rDNA. Sequencing data showed on a genus level, an abundance of Lactobacillus, Streptococcus, and Clostridium XI (38.2, 28.1, and 13.0% respectively) irrespective of location and age. The caeca exhibited the greatest microbiome diversity throughout the development of the turkey. PICRUSt data predicted an array of bacterial function, with most differences being apparent in the caeca of the turkeys as they matured. QPCR revealed that the caeca within 10 week old birds, contained the most Campylobacter spp. Understanding the microbial ecology of the turkey gastrointestinal tract is essential in terms of understanding production efficiency and in order to develop novel strategies for targeting Campylobacter spp.


Laparoscopic Adrenalectomy for Large Adrenocortical Carcinoma.
Adrenocortical cancer (ACC) is a rare disease that is difficult to treat. Laparoscopic adrenalectomy (LA) is performed, even for large adrenocortical carcinomas. However, the oncological effectiveness of LA remains unclear. This review presents the current knowledge of the feasibility and oncological effectiveness of laparoscopic surgery for ACC, with an analysis of data for outcomes and other parameters. A systematic review of the literature was performed by searching the PubMed and Medline databases for all relevant articles in English, published between January 1992 and August 2014 on LA for adrenocortical carcinoma. The search resulted in retrieval of 29 studies, of which 10 addressed the outcome of LA versus open adrenalectomy (OA) and included 844 patients eligible for this review. Among these, 206 patients had undergone LA approaches, and 638 patients had undergone OA. Among the 10 studies that compared the outcomes obtained with LA and OA for ACC, 5 noted no statistically significant difference between the 2 groups in the oncological outcomes of recurrence and disease-free survival, whereas the remaining 5 reported inferior outcomes in the LA group. Using a paired t test for statistical analysis, except for tumor size, we found no significant difference in local recurrence, peritoneal carcinomatosis, positive resection margin, and time to recurrence between the LA and OA groups. The overall mean tumor size in patients undergoing LA and OA was 7.1 and 11.2 cm, respectively (P = .0003), and the mean overall recurrence was 61.5 and 57.9%, respectively. The outcome of LA is believed to depend to a large extent on the size and stage of the lesion (I and II being favorable) and the surgical expertise in the center where the patient undergoes the operation. However, the present review shows no difference in the outcome between the 2 approaches across all stages. A poor outcome is likely to result from inadequate surgery, irrespective of whether the approach is open or laparoscopic.


Structures of N-Glycans of Bothrops Venoms Revealed as Molecular Signatures that Contribute to Venom Phenotype in Viperid Snakes.
The complexity of snake venoms has long been investigated to explore a myriad of biologically active proteins and peptides that are used for immobilizing or killing prey, and are responsible for the pathological effects observed on envenomation. Glycosylation is the main post-translational modification (PTM) of viperid venoms but currently there is little understanding of how protein glycosylation impacts the variation of venom proteomes. We have previously reported that Bothrops venom glycoproteomes contain a core of components that markedly define their composition and parallel their phylogenetic classification. Here we extend those observations to eight Bothrops species evaluating the N-glycomes by LC-MS as assigned cartoon structures and detailing those structures separately as methylated analogs using ion-trap mass spectrometry (MSn). Following ion disassembly through multiple steps provided sequence and linkage isomeric details that characterized 52 unique compositions in Bothrops venoms. These occurred as 60 structures, of which 26 were identified in the venoms of the Jararaca Complex (B. alcatraz, B. insularis, and B. jararaca), 20 in B. erythromelas, B. jararacussu, B. moojeni and B. neuwiedi venoms, and 22 in B. cotiara venom. Further, quantitative analysis of these N-glycans showed variable relative abundances in the venoms. For the first time a comprehensive set of N-glycan structures present in snake venoms are defined. Despite the fact that glycosylation is not template-defined, the N-glycomes of these venoms mirror the phylogeny cladograms of South American bothropoid snakes reported in studies on morphological, molecular data and feeding habits, exhibiting distinct molecular signatures for each venom. Considering the complexity of N-glycan moieties generally found in glycoproteins, characterized by different degrees of branching, isomer structures, and variable abundances, our findings point to these factors as another level of complexity in Bothrops venoms, features that could dramatically contribute to their distinct biological activities.


Human exercise-mediated skeletal muscle hypertrophy is an intrinsic process.
Muscle cells (fibres) are post-mitotic and thus undergo changes in phenotype by modifying their existing structure. Hypertrophy is a hallmark change that occurs in response to increased loading and can be achieved in humans through repeated bouts of resistance exercise (i.e., training). In resistance exercise, contractions are initiated by neural drive leading to immediate perturbations such as calcium influx, cross-bridge cycling and tension/stress on the cytoskeleton, sarcolemma and extracellular matrix, as well as more delayed cellular events such as the production/release of potential local growth factors (e.g., IGF-1). Resistance exercise can also elevate the systemic concentration of certain hormones (growth hormone, testosterone, IGF-1) that are hypothesized to drive hypertrophy. However, while these hormones are clearly anabolic during childhood and puberty, or when given at supraphysiological exogenous doses, the transient post-exercise elevations in hormone concentration are of little consequence to the either the acute protein synthetic response or to a hypertrophic phenotype after resistance training. Thus, the acute post-exercise increases in systemic hormones are in no way a proxy marker for anabolism since they do not underpin the capacity of the muscle to hypertrophy in any measurable way. In contrast, the acute activation of intrinsically located signalling proteins such as p70(S6K) and the acute elevation of muscle protein synthesis are more reflective of the potential to increase in muscle mass with resistance training. Ultimately, local mechanisms are activated by the stress imposed by muscle loading and prime the muscle for protein accretion. Membrane-derived molecules and tension-sensing pathways are two intrinsic mechanisms implicated in upregulating the synthesis and incorporation of muscle proteins into the myofibre in response to mechanical stress derived from loaded contractions.


Prospective associations of fasting insulin, body fat distribution, and diabetes with risk of ischemic stroke. The Atherosclerosis Risk in Communities (ARIC) Study Investigators.
We tested the hypothesis that diabetes, body fat distribution, and (in nondiabetic subjects) fasting insulin levels are positively associated with ischemic stroke incidence in the general population. As part of the Atherosclerosis Risk in Communities (ARIC) Study, we measured diabetes by using fasting glucose criteria, waist and hip circumferences, and fasting insulin levels with a radioimmunoassay in > 12,000 adults aged 45-64 years who had no cardiovascular disease at baseline. We followed them for 6-8 years for ischemic stroke occurrence (n = 191). After adjustment for age, sex, race, ARIC community, smoking, and education level, the relative risk of ischemic stroke was 3.70 (95% CI 2.7-5.1) for diabetes, 1.74 (1.4-2.2) for a 0.11 increment of waist-to-hip ratio, and 1.19 (1.1-1.3) for a 50-pmol/l increment of fasting insulin among nondiabetic subjects. Ischemic stroke incidence was not statistically significantly associated with BMI (comparably adjusted relative risk = 1.15, 95% CI 0.97-1.36). With adjustment for other stroke risk factors (some of which may mediate the effects of diabetes, fat distribution, and hyperinsulinemia), the relative risks for diabetes, waist-to-hip ratio, and fasting insulin level were 2.22 (95% CI 1.5-3.2), 1.08 (0.8-1.4), and 1.14 (1.01-1.3), respectively. Diabetes is a strong risk factor for ischemic stroke. Aspects of insulin resistance, as reflected by elevated waist-to-hip ratios and elevated fasting insulin levels, may also contribute to a greater risk of ischemic stroke.


Identification of the Molecular Clockwork of the Oyster Crassostrea gigas.
Molecular clock system constitutes the origin of biological rhythms that allow organisms to anticipate cyclic environmental changes and adapt their behavior and physiology. Components of the molecular clock are largely conserved across a broad range of species but appreciable diversity in clock structure and function is also present especially in invertebrates. The present work aimed at identify and characterize molecular clockwork components in relationship with the monitoring of valve activity behavior in the oyster Crassostrea gigas. Results provided the characterization of most of canonical clock gene including clock, bmal/cycle, period, timeless, vertebrate-type cry, rev-erb, ror as well as other members of the cryptochrome/photolyase family (plant-like cry, 6-4 photolyase). Analyses of transcriptional variations of clock candidates in oysters exposed to light / dark regime and to constant darkness led to the generation of a putative and original clockwork model in C. gigas, intermediate of described systems in vertebrates and insects. This study is the first characterization of a mollusk clockwork. It constitutes essential bases to understand interactions of the different components of the molecular clock in C. gigas as well as the global mechanisms associated to the generation and the synchronization of biological rhythms in oysters.