Human rotavirus genotypes causing acute watery diarrhea among under-five children in Benin City, Nigeria.
Diarrhea is a major cause of childhood morbidity and mortality in the developing countries. Rotavirus is a major cause of acute watery diarrhea. This study aims at characterizing the prevalent rotavirus G-genotypes among under-five children presenting with acute watery diarrhea in Benin City, Nigeria. A total of 470 children <5 years presenting with diarrhea of <2 weeks duration, were over a period of 1 year consecutively recruited for the study. Stool samples were collected for rotavirus antigen detection using Enzyme-Linked Immunosorbent Assay (ELISA) and further analyzed with reverse transcriptase polymerase chain reaction (RT-PCR) for VP7 genotyping. Comparing the ability of the two methods to detect rotavirus in stool samples, 65 (13.8%) and 90 (19.2%) of the stools tested positive for rotavirus using ELISA and RT-PCR, respectively. Using VP7 primers, genotypes G1 were detected in 49 out of 90 stool samples (54.4%), G2 in 26 out of 90 stool samples (28.9%), G3 in 19 out of 90 stool samples (21.1%), G4 in 34 out of 90 stool samples (37.8%) and G9 in 8 of the 90 stool samples (8.9%). Some strains were observed to be reactive with 2 or more of the primers yielding dual or triple VP7 genotype reactivity. Rotavirus of varying genotypes as shown cause acute watery diarrhea among under-five children and vaccine with strains peculiar to this environment should be introduced. Techniques such as RT-PCR rather than ELISA, where affordable, should be used in stool rotavirus screening.


Male sex workers: practices, contexts, and vulnerabilities for HIV acquisition and transmission.
Male sex workers who sell or exchange sex for money or goods encompass a very diverse population across and within countries worldwide. Information characterising their practices, contexts where they live, and their needs is limited, because these individuals are generally included as a subset of larger studies focused on gay men and other men who have sex with men (MSM) or even female sex workers. Male sex workers, irrespective of their sexual orientation, mostly offer sex to men and rarely identify as sex workers, using local or international terms instead. Growing evidence indicates a sustained or increasing burden of HIV among some male sex workers within the context of the slowing global HIV pandemic. Several synergistic facilitators could be potentiating HIV acquisition and transmission among male sex workers, including biological, behavioural, and structural determinants. Criminalisation and intersectional stigmas of same-sex practices, commercial sex, and HIV all augment risk for HIV and sexually transmitted infections among male sex workers and reduce the likelihood of these people accessing essential services. These contexts, taken together with complex sexual networks among male sex workers, define this group as a key population underserved by current HIV prevention, treatment, and care services. Dedicated efforts are needed to make those services available for the sake of both public health and human rights. Evidence-based and human rights-affirming services dedicated specifically to male sex workers are needed to improve health outcomes for these men and the people within their sexual networks.


Identifying bedrest using waist-worn triaxial accelerometers in preschool children.
To adapt and validate a previously developed decision tree for youth to identify bedrest for use in preschool children. Parents of healthy preschool (3-6-year-old) children (n = 610; 294 males) were asked to help them to wear an accelerometer for 7 to 10 days and 24 hours/day on their waist. Children with ≥3 nights of valid recordings were randomly allocated to the development (n = 200) and validation (n = 200) groups. Wear periods from accelerometer recordings were identified minute-by-minute as bedrest or wake using visual identification by two independent raters. To automate visual identification, chosen decision tree (DT) parameters (block length, threshold, bedrest-start trigger, and bedrest-end trigger) were optimized in the development group using a Nelder-Mead simplex optimization method, which maximized the accuracy of DT-identified bedrest in 1-min epochs against synchronized visually identified bedrest (n = 4,730,734). DT's performance with optimized parameters was compared with the visual identification, commonly used Sadeh's sleep detection algorithm, DT for youth (10-18-years-old), and parental survey of sleep duration in the validation group. On average, children wore an accelerometer for 8.3 days and 20.8 hours/day. Comparing the DT-identified bedrest with visual identification in the validation group yielded sensitivity = 0.941, specificity = 0.974, and accuracy = 0.956. The optimal block length was 36 min, the threshold 230 counts/min, the bedrest-start trigger 305 counts/min, and the bedrest-end trigger 1,129 counts/min. In the validation group, DT identified bedrest with greater accuracy than Sadeh's algorithm (0.956 and 0.902) and DT for youth (0.956 and 0.861) (both P<0.001). Both DT (564±77 min/day) and Sadeh's algorithm (604±80 min/day) identified significantly less bedrest/sleep than parental survey (650±81 min/day) (both P<0.001). The DT-based algorithm initially developed for youth was adapted for preschool children to identify time spent in bedrest with high accuracy. The DT is available as a package for the R open-source software environment ("PhysActBedRest").


The role of environmental antigens in the spontaneous development of autoimmunity in MRL-lpr mice.
It has been proposed that the "normal" stimulation of the immune system that occurs from interactions with environmental stimuli, whether infectious or dietary, is necessary for the initiation and/or continuation of autoimmunity. We tested this hypothesis by deriving a group of MRL-lpr mice into a germfree (GF) environment. At 5 mo of age, no differences between GF and conventional MRL-lpr mice were noted in lymphoproliferation, flow cytometric analysis of lymph node cells (LN), or histologic analysis of the kidneys. Autoantibody levels were comparably elevated in both groups. A second experiment tested the role of residual environmental stimuli by contrasting GF mice fed either a low m.w., ultrafiltered Ag-free (GF-AF) diet or an autoclaved natural ingredient diet (GF-NI). At 4 mo of age, both groups showed extensive lymphoproliferation and aberrant T cell formation, although the GF-AF mice had approximately 50% smaller LNs compared with sex-matched GF-NI controls. Autoantibody formation was present in both groups. Histologic analysis of the kidneys revealed that GF-AF mice had much lower levels of nephritis, while immunofluorescence analysis demonstrated no difference in Ig deposits but did reveal a paucity of C3 deposition in the kidneys of GF-AF mice. These data do not support a role for infectious agents in the induction of lymphoproliferation and B cell autoimmunity in MRL-lpr mice. Furthermore, they suggest that autoantibodies do not originate from B cells that were initially committed to exogenous Ags. They do suggest a possible contributory role for dietary exposure in the extent of lymphoproliferation and development of nephritis in this strain.


Personal circumstances and social characteristics as determinants of landholder participation in biodiversity conservation programs.
Adequate conservation of biodiversity on private land remains elusive due, in part, to a failure to understand the personal circumstances and social characteristics of private landholders. Our aim was to identify those personal and social dimensions of landholders that might contribute to improved conservation policy and program design and, thereby, participation in private land conservation. We tested whether personal circumstances of landholders (e.g., lifestyle and wellbeing, information and knowledge, financial security) and social characteristics (e.g., attitudes, norms, and trust) would be important predictors of landholders' capacity and willingness to participate in biodiversity conservation programs. Forty-five participants and twenty-nine non-participants of biodiversity conservation programs in north Queensland, Australia, were surveyed to: 1) examine differences between their personal circumstances and social characteristics that may influence participation; and 2) explore whether personal circumstances and social characteristics were influenced by participation. The results revealed that, compared to participants, non-participants in conservation programs had significantly different personal circumstances and social characteristics for four of eight measured variables. Compared to participants, non-participants demonstrated a reduced capacity and willingness to participate in conservation programs. Participation did not appear to have a strong influence on participants' personal circumstances or social characteristics, and when social norms supported conservation, programs did not demonstrate additionality. Conservation policies that maintain or improve landholders' personal circumstances and that promote pro-environmental norms may result in increased participation and thereby conservation outcomes.


Medical history, lifestyle, family history, and occupational risk factors for mantle cell lymphoma: the InterLymph Non-Hodgkin Lymphoma Subtypes Project.
The etiology of mantle cell lymphoma (MCL), a distinctive subtype accounting for 2%-10% of all non-Hodgkin lymphoma, is not known. We investigated associations with self-reported medical history, lifestyle, family history, and occupational risk factors in a pooled analysis of 557 patients with MCL and 13766 controls from 13 case-control studies in Europe, North America, and Australia. Odds ratios (ORs) and 95% confidence intervals (CIs) associated with each exposure were examined using multivariate logistic regression models. The median age of the MCL patients was 62 years and 76% were men. Risk of MCL was inversely associated with history of hay fever (OR = 0.63, 95% CI = 0.48 to 0.82), and the association was independent of other atopic diseases and allergies. A hematological malignancy among first-degree relatives was associated with a twofold increased risk of MCL (OR = 1.99, 95% CI = 1.39 to 2.84), which was stronger in men (OR = 2.21, 95% CI = 1.44 to 3.38) than women (OR = 1.61, 95% CI = 0.82 to 3.19). A modestly increased risk of MCL was also observed in association with ever having lived on a farm (OR = 1.40, 95% CI = 1.03 to 1.90). Unlike some other non-Hodgkin lymphoma subtypes, MCL risk was not statistically significantly associated with autoimmune disorders, tobacco smoking, alcohol intake, body mass index, or ultraviolet radiation. The novel observations of a possible role for atopy and allergy and farm life in risk of MCL, together with confirmatory evidence of a familial link, suggest a multifactorial etiology of immune-related environmental exposures and genetic susceptibility. These findings provide guidance for future research in MCL etiology.


Intravenous Air: The Partially Invisible Phenomenon.
Air injection is carefully avoided during IV solution administration; however, ambient air is dissolved in all liquids used for intravenous (IV) therapy. A portion of this gas will come out of solution in the form of bubbles as the solution is warmed to body temperature in a fluid warming system and/or within the body. We sought to quantify the proportion of the gas theoretically dissolved in room temperature crystalloid and 4°C blood products that comes out of solution in the IV tubing on warming to 37°C. Equilibrium-dissolved air calculations were performed for sodium chloride (0.9%), packed red blood cells, and fresh frozen plasma at various temperatures according to Henry's Law. Outgassed gas volumes were experimentally measured for room temperature sodium chloride (0.9%) and 4°C blood products (packed red blood cells and fresh frozen plasma) warmed to 37°C during infusion into a body temperature water bath. The measured gas volumes were quantified as a fraction of the theoretical outgassing volumes required to maintain equilibrium saturation. Measured outgassed volumes in the IV tubing in milliliters of gas per liter of fluid were 1.4 ± 0.3 mL/L (n = 6) for sodium chloride (0.9%), 3.4 ± 0.2 mL/L (n = 6) for packed red blood cells, and 4.8 ± 0.8 mL/L (n = 6) for fresh frozen plasma when these fluids were warmed to body temperature from their respective starting temperatures. Theoretical outgassed gas volumes required to maintain equilibrium saturation for the same fluids and temperatures are 4.7 mL/L for sodium chloride (0.9%), 8.3 mL/L for packed red blood cells, and 10.9 mL/L for fresh frozen plasma. As a fraction of the theoretical outgassing volumes, the measured air volumes represented 30%, 41%, and 44%, respectively, for sodium chloride (0.9%), packed red blood cells, and fresh frozen plasma. Prewarming crystalloid solutions to 37°C before administration significantly reduced the outgassing. A significant and potentially clinically relevant amount of the resident dissolved gas in room temperature crystalloid, and 4°C packed red blood cells and plasma solutions comes out of solution on warming to body temperature. A nontrivial fraction of this outgassing is also expected to occur within the body circulation based on the results of this study. This can be substantially prevented by prewarming.


The development and application of bioinformatics core competencies to improve bioinformatics training and education.
Bioinformatics is recognized as part of the essential knowledge base of numerous career paths in biomedical research and healthcare. However, there is little agreement in the field over what that knowledge entails or how best to provide it. These disagreements are compounded by the wide range of populations in need of bioinformatics training, with divergent prior backgrounds and intended application areas. The Curriculum Task Force of the International Society of Computational Biology (ISCB) Education Committee has sought to provide a framework for training needs and curricula in terms of a set of bioinformatics core competencies that cut across many user personas and training programs. The initial competencies developed based on surveys of employers and training programs have since been refined through a multiyear process of community engagement. This report describes the current status of the competencies and presents a series of use cases illustrating how they are being applied in diverse training contexts. These use cases are intended to demonstrate how others can make use of the competencies and engage in the process of their continuing refinement and application. The report concludes with a consideration of remaining challenges and future plans.


The denaturation and degradation of stable enzymes at high temperatures.
Now that enzymes are available that are stable above 100 degrees C it is possible to investigate conformational stability at this temperature, and also the effect of high-temperature degradative reactions in functioning enzymes and the inter-relationship between degradation and denaturation. The conformational stability of proteins depends upon stabilizing forces arising from a large number of weak interactions, which are opposed by an almost equally large destabilizing force due mostly to conformational entropy. The difference between these, the net free energy of stabilization, is relatively small, equivalent to a few interactions. The enhanced stability of very stable proteins can be achieved by an additional stabilizing force which is again equivalent to only a few stabilizing interactions. There is currently no strong evidence that any particular interaction (e.g. hydrogen bonds, hydrophobic interactions) plays a more important role in proteins that are stable at 100 degrees C than in those stable at 50 degrees C, or that the structures of very stable proteins are systematically different from those of less stable proteins. The major degradative mechanisms are deamidation of asparagine and glutamine, and succinamide formation at aspartate and glutamate leading to peptide bond hydrolysis. In addition to being temperature-dependent, these reactions are strongly dependent upon the conformational freedom of the susceptible amino acid residues. Evidence is accumulating which suggests that even at 100 degrees C deamidation and succinamide formation proceed slowly or not at all in conformationally intact (native) enzymes. Whether this is the case at higher temperatures is not yet clear, so it is not known whether denaturation of degradation will set the upper limit of stability for enzymes.


Mechanisms of cell death induced by the mitochondrial toxin 3-nitropropionic acid: acute excitotoxic necrosis and delayed apoptosis.
Impaired energy metabolism may play an important role in neuronal cell death after brain ischemia and in late-onset neurodegenerative diseases. Both excitotoxic necrosis and apoptosis have been implicated in cell death induced by metabolic impairment. However, the factors that determine whether cells undergo apoptosis or necrosis are not known. In the present study, metabolic impairment was induced by 3-nitropropionic acid (3-NP), a suicide inhibitor of succinate dehydrogenase. Treatment of cultured rat hippocampal neurons with 3-NP resulted in two types of cell death with distinct morphological, pharmacological, and biochemical features. A rapid necrotic cell death, characterized by cell swelling and nuclear shrinkage, could be completely prevented by the NMDA receptor antagonist MK-801 (10 microM) and dose-dependently potentiated by low micromolar levels of extracellular glutamate. A slowly evolving apoptotic death, characterized by nuclear fragmentation, was not attenuated by MK-801 but was prevented by cycloheximide (1 microg/ml). The combination of MK-801 and cycloheximide resulted in an almost complete protection against 3-NP-induced cell death. DNA fragmentation, detected by the terminal deoxynucleotidyl transferase-mediated dUTP-X 3' nick end-labeling technique, was a late event in apoptosis and also occurred after necrotic cell death. ATP depletion was an early event in the 3-NP-induced neuronal degeneration, and the decline in ATP was exacerbated by glutamate. We conclude that 3-NP triggers two separate cell death pathways: an excitotoxic necrosis as a result of NMDA receptor activation and a delayed apoptosis that is NMDA receptor-independent. Mildly elevated levels of extracellular glutamate shift the cell death mechanism from apoptosis to necrosis.


Atrial fibrillation is associated with high levels of monocyte-platelet-aggregates and increased CD11b expression in patients with aortic stenosis.
A growing body of evidence suggests a pivotal role of inflammatory processes in AF in a bidirectional manner. Infiltrating leukocytes seem to promote both structural and electrical remodelling processes in patients with AF. Monocyte-platelets-aggregates (MPAs) are sensitive markers of both platelets and monocyte activation. So far it is not clear whether the content of MPAs is affected by AF. The present study examined the content of MPAs and the activation of monocytes in elderly patients with an aortic stenosis in dependence of AF. These patients are known to have a high prevalence of AF. Flow-cytometric quantification analysis demonstrated that patients with AF have an increased content of MPAs (207 ± 13 cells/µl vs 307 ± 21 cells/µl, p< 0.001), and enhanced expression of CD11b on monocytes (p< 0.001), compared to patients in stable sinus rhythm (SR). The number of CD14+/CD16+ monocytes were only slightly elevated in patients with AF. These findings were seen in patients with permanent AF. But also patients with paroxysmal AF, even when presenting in SR, the MPAs were increased by 50 % (p< 0.05) as well as the CD11b expression, which was twice as high (p< 0.05) compared to stable SR. These results demonstrate for the first time a dependency of MPAs and CD11b expression on monocytes in the presence of AF and support the notion of a close relationship between AF, thrombogenesis and inflammation. The content of MPAs and the extent of activation on monocytes appear promising as biomarkers for paroxysmal AF and as possible future targets for developing novel pharmacological therapeutic strategies.


Hepatitis E Virus (HEV)-Specific T Cell Receptor Cross-Recognition: Implications for Immunotherapy.
T cell immunotherapy is a concept developed for the treatment of cancer and infectious diseases, based on cytotoxic T lymphocytes to target tumor- or pathogen-specific antigens. Antigen-specificity of the T cell receptors (TCRs) is an important selection criterion in the developmental design of immunotherapy. However, off-target specificity is a possible autoimmunity concern if the engineered antigen-specific T cells are cross-reacting to self-peptides in-vivo. In our recent work, we identified several hepatitis E virus (HEV)-specific TCRs as potential candidates to be developed into T cell therapy to treat chronic hepatitis E. One of the identified TCRs, targeting a HLA-A2-restricted epitope at the RNA-dependent RNA polymerase (HEV-1527: LLWNTVWNM), possessed a unique multiple glycine motif in the TCR-β CDR3, which might be a factor inducing cross-reactivity. The aim of our study was to explore if this TCR could cross-recognize self-peptides to underlay autoimmunity. Indeed, we found that this HEV-1527-specific TCR could also cross-recognize an apoptosis-related epitope, Nonmuscle Myosin Heavy Chain 9 (MYH9-478: QLFNHTMFI). While this TCR had dual specificities to both viral epitope and a self-antigen by double Dextramer binding, it was selectively functional against HEV-1527 but not activated against MYH9-478. The consecutive glycine motif in β chain may be the reason promoting TCR binding promiscuity to recognize a secondary target, thereby facilitating cross-recognition. In conclusion, candidate TCRs for immunotherapy development should be screened for autoimmune potential, especially when the TCRs exhibit unique sequence pattern.


Updated Guidelines for the Medical Assessment and Care of Children Who May Have Been Sexually Abused.
The medical evaluation is an important part of the clinical and legal process when child sexual abuse is suspected. Practitioners who examine children need to be up to date on current recommendations regarding when, how, and by whom these evaluations should be conducted, as well as how the medical findings should be interpreted. A previously published article on guidelines for medical care for sexually abused children has been widely used by physicians, nurses, and nurse practitioners to inform practice guidelines in this field. Since 2007, when the article was published, new research has suggested changes in some of the guidelines and in the table that lists medical and laboratory findings in children evaluated for suspected sexual abuse and suggests how these findings should be interpreted with respect to sexual abuse. A group of specialists in child abuse pediatrics met in person and via online communication from 2011 through 2014 to review published research as well as recommendations from the Centers for Disease Control and Prevention and the American Academy of Pediatrics and to reach consensus on if and how the guidelines and approach to interpretation table should be updated. The revisions are based, when possible, on data from well-designed, unbiased studies published in high-ranking, peer-reviewed, scientific journals that were reviewed and vetted by the authors. When such studies were not available, recommendations were based on expert consensus.


Modeling Perturbations in Protein Filaments at the Micro and Meso Scale Using NAMD and PTools/Heligeom.
Protein filaments are dynamic entities that respond to external stimuli by slightly or substantially modifying the internal binding geometries between successive protomers. This results in overall changes in the filament architecture, which are difficult to model due to the helical character of the system. Here, we describe how distortions in RecA nucleofilaments and their consequences on the filament-DNA and bound DNA-DNA interactions at different stages of the homologous recombination process can be modeled using the PTools/Heligeom software and subsequent molecular dynamics simulation with NAMD. Modeling methods dealing with helical macromolecular objects typically rely on symmetric assemblies and take advantage of known symmetry descriptors. Other methods dealing with single objects, such as MMTK or VMD, do not integrate the specificities of regular assemblies. By basing the model building on binding geometries at the protomer-protomer level, PTools/Heligeom frees the building process from a priori knowledge of the system topology and enables irregular architectures and symmetry disruption to be accounted for. Graphical abstract: Model of ATP hydrolysis-induced distortions in the recombinant nucleoprotein, obtained by combining RecA-DNA and two RecA-RecA binding geometries.


An electric-field-active 1377-nm narrow-line Si light-emitting diode at 150 K.
A new class of silicon-based light-emitting diode is demonstrated using InSb-quantum-dot-embedded Si containing the emissive {311} rod-like defects (RLDs). A narrow peak centered at 1377 nm (900 meV) characteristic of the {311} RLDs was found to develop out of an otherwise broad background electroluminescence (EL) upon the application of electric fields in the growth direction. Such electric-field-active EL was observed up to 150 K with a slight downward shift of the peak energies, accompanied by an anomaly in the thermal roll-off of the EL intensity. Spectral variations with temperature and electric field indicate a switching of dominance between the closely correlated defect states that are responsible for the EL emission.


Using conversation topics for predicting therapy outcomes in schizophrenia.
Previous research shows that aspects of doctor-patient communication in therapy can predict patient symptoms, satisfaction and future adherence to treatment (a significant problem with conditions such as schizophrenia). However, automatic prediction has so far shown success only when based on low-level lexical features, and it is unclear how well these can generalize to new data, or whether their effectiveness is due to their capturing aspects of style, structure or content. Here, we examine the use of topic as a higher-level measure of content, more likely to generalize and to have more explanatory power. Investigations show that while topics predict some important factors such as patient satisfaction and ratings of therapy quality, they lack the full predictive power of lower-level features. For some factors, unsupervised methods produce models comparable to manual annotation.


Human cytomegalovirus infection of bone marrow myofibroblasts enhances myeloid progenitor adhesion and elicits viral transmission.
Human cytomegalovirus (CMV) infection of bone marrow transplant recipients can cause pancytopenia, as well as life-threatening interstitial pneumonia. CMV replicates actively in bone marrow stromal cells, whereas it remains latent in hematopoietic progenitors. Our aim was to study the influence of CMV infection on adherence of CD34(+) cells to the myofibroblastic component of human bone marrow and examine transmission of virus from myofibroblasts to CD34(+) cells. We show that smooth actin, but not fibronectin, organization is markedly modified by CMV infection of bone marrow stromal myofibroblasts. Nonetheless, CMV infection led to increased adherence of the CD34(+) progenitor cell line, KG1a, relative to adherence to uninfected myofibroblasts from the same donors. Adherence of CD34(+) cells to infected bone marrow myofibroblasts resulted in transfer of virions and viral proteins through close cell-to-cell contacts. This phenomenon may play a role in the pathophysiology of CMV bone marrow infection and in eventual virus dissemination.


Simplified and efficient quantification of low-abundance proteins at very high multiplex via targeted mass spectrometry.
Liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM-MS) of plasma that has been depleted of abundant proteins and fractionated at the peptide level into six to eight fractions is a proven method for quantifying proteins present at low nanogram-per-milliliter levels. A drawback of fraction-MRM is the increased analysis time due to the generation of multiple fractions per biological sample. We now report that the use of heated, long, fused silica columns (>30 cm) packed with 1.9 μm of packing material can reduce or eliminate the need for fractionation prior to LC-MRM-MS without a significant loss of sensitivity or precision relative to fraction-MRM. We empirically determined the optimal column length, temperature, gradient duration, and sample load for such assays and used these conditions to study detection sensitivity and assay precision. In addition to increased peak capacity, longer columns packed with smaller beads tolerated a 4- to 6-fold increase in analyte load without a loss of robustness or reproducibility. The longer columns also provided a 4-fold improvement in median limit-of-quantitation values with increased assay precision relative to the standard 12 cm columns packed with 3 μm material. Overall, the optimized chromatography provided an approximately 3-fold increase in analysis throughput with excellent robustness and less than a 2-fold reduction in quantitative sensitivity relative to fraction-MRM. The value of the system for increased multiplexing was demonstrated by the ability to configure an 800-plex MRM-MS assay, run in a single analysis, comprising 2400 transitions with retention time scheduling to monitor 400 unlabeled and heavy labeled peptide pairs.


Eidinemacheilus proudlovei, a new subterranean loach from Iraqi Kurdistan (Teleostei; Nemacheilidae).
Eidinemacheilus proudlovei, new species, is described from subterranean waters in the Little Zab River drainage in Iraqi Kurdistan. After the discovery of E. smithi in 1976, E. proudlovei is the second troglomorphic nemacheilid loach found in the Middle East and the second species placed in Eidinemacheilus. Eidinemacheilus proudlovei is distinguished from E. smithi by having 8+8 or 8+7 branched caudal-fin rays, no adipose keel on the caudal peduncle, enlarged jaws and a fully developed head canal system. It furthers differs substantially in its DNA barcode (>8% K2P distance) from all other nemacheilid loaches in the Middle East, Europe and Western India.


The secretory proprotein convertases furin, PC5, and PC7 activate VEGF-C to induce tumorigenesis.
The secretory factor VEGF-C has been directly implicated in various physiological processes during embryogenesis and human cancers. However, the importance of the conversion of its precursor proVEGF-C to mature VEGF-C in tumorigenesis, and vessel formation and the identity of the protease(s) that regulate these processes is/are not known. The intracellular processing of proVEGF-C that occurs within the dibasic motif HSIIRR(227)SL suggests the involvement of the proprotein convertases (PCs) in this process. In addition, furin and VEGF-C were found to be coordinately expressed in adult mouse tissues. Cotransfection of the furin-deficient colon carcinoma cell line LoVo with proVEGF-C and different PC members revealed that furin, PC5, and PC7 are candidate VEGF-C convertases. This finding is consistent with the in vitro digestions of an internally quenched synthetic fluorogenic peptide mimicking the cleavage site of proVEGF-C ((220)Q-VHSIIRR downward arrow SLP(230)). The processing of proVEGF-C is blocked by the inhibitory prosegments of furin, PC5, and PACE4, as well as by furin-motif variants of alpha2-macroglobulin and alpha1-antitrypsin. Subcutaneous injection of CHO cells stably expressing VEGF-C into nude mice enhanced angiogenesis and lymphangiogenesis, but not tumor growth. In contrast, expression of proVEGF-C obtained following mutation of the cleavage site (HSIIRR(227)SL to HSIISS(227)SL) inhibits angiogenesis and lymphangiogenesis as well as tumor growth. Our findings demonstrate the processing of proVEGF-C by PCs and highlight the potential use of PC inhibitors as agents for inhibiting malignancies induced by VEGF-C.


Uncertainty and sensitivity analysis for reducing greenhouse gas emissions from wastewater treatment plants.
This paper presents the sensitivity and uncertainty analysis of a plant-wide mathematical model for wastewater treatment plants (WWTPs). The mathematical model assesses direct and indirect (due to the energy consumption) greenhouse gases (GHG) emissions from a WWTP employing a whole-plant approach. The model includes: (i) the kinetic/mass-balance based model regarding nitrogen; (ii) two-step nitrification process; (iii) N2O formation both during nitrification and denitrification (as dissolved and off-gas concentration). Important model factors have been selected by using the Extended-Fourier Amplitude Sensitivity Testing (FAST) global sensitivity analysis method. A scenario analysis has been performed in order to evaluate the uncertainty related to all selected important model factors (scenario 1), important model factors related to the influent features (scenario 2) and important model factors related to the operational conditions (scenario 3). The main objective of this paper was to analyse the key factors and sources of uncertainty at a plant-wide scale influencing the most relevant model outputs: direct and indirect (DIR,CO2eq and IND,CO2eq, respectively), effluent quality index (EQI), chemical oxygen demand (COD) and total nitrogen (TN) effluent concentration (CODOUT and TNOUT, respectively). Sensitivity analysis shows that model factors related to the influent wastewater and primary effluent COD fractionation exhibit a significant impact on direct, indirect and EQI model factors. Uncertainty analysis reveals that outflow TNOUT has the highest uncertainty in terms of relative uncertainty band for scenario 1 and scenario 2. Therefore, uncertainty of influential model factors and influent fractionation factors has a relevant role on total nitrogen prediction. Results of the uncertainty analysis show that the uncertainty of model prediction decreases after fixing stoichiometric/kinetic model factors.


Practices for Identifying and Rejecting Hemolyzed Specimens Are Highly Variable in Clinical Laboratories.
Hemolysis is an important clinical laboratory quality attribute that influences result reliability. To determine hemolysis identification and rejection practices occurring in clinical laboratories. We used the College of American Pathologists Survey program to distribute a Q-Probes-type questionnaire about hemolysis practices to Chemistry Survey participants. Of 3495 participants sent the questionnaire, 846 (24%) responded. In 71% of 772 laboratories, the hemolysis rate was less than 3.0%, whereas in 5%, it was 6.0% or greater. A visual scale, an instrument scale, and combination of visual and instrument scales were used to identify hemolysis in 48%, 11%, and 41% of laboratories, respectively. A picture of the hemolysis level was used as an aid to technologists' visual interpretation of hemolysis levels in 40% of laboratories. In 7.0% of laboratories, all hemolyzed specimens were rejected; in 4% of laboratories, no hemolyzed specimens were rejected; and in 88% of laboratories, some specimens were rejected depending on hemolysis levels. Participants used 69 different terms to describe hemolysis scales, with 21 terms used in more than 10 laboratories. Slight and moderate were the terms used most commonly. Of 16 different cutoffs used to reject hemolyzed specimens, moderate was the most common, occurring in 30% of laboratories. For whole blood electrolyte measurements performed in 86 laboratories, 57% did not evaluate the presence of hemolysis, but for those that did, the most common practice in 21 laboratories (24%) was centrifuging and visually determining the presence of hemolysis in all specimens. Hemolysis practices vary widely. Standard assessment and consistent reporting are the first steps in reducing interlaboratory variability among results.


Quantitative risk assessment for developmental toxicity.
Pharmaceutical companies and governmental regulatory agencies are becoming increasingly aware of the need for improved statistical methods for developmental toxicity experiments. Although a number of statisticians have become interested in this area, activity has centered mostly on the development of methods to analyze binary outcomes, such as malformations among live pups, while accounting appropriately for the correlation induced by the litter effect. In contrast, the topic of quantitative risk assessment has received relatively little attention. This paper addresses the specific question of how to assess risk appropriately when exposure causes a variety of adverse effects, including resorption and fetal death, in addition to malformations. It will be seen that risk assessments based on a single developmental outcome, such as malformation, may be conservative. A method is proposed for estimating an exposure level at which the overall risk of any adverse effect is acceptably low. The method is based on a continuation ratio formulation of a multinomial distribution, with an additional scale parameter to account for overdispersion. Comparisons are made with binary models on prenatal death and malformation, as well as a binary model that makes no distinction between death and malformation, but simply classifies each fetus as normal or abnormal. Data from several developmental toxicity studies illustrate the results and findings.


Persistence analysis of the static and dynamical helix deformations of DNA oligonucleotides: application to the crystal structure and molecular dynamics simulation of d(CGCGAATTCGCG)2.
A theory and graphical presentation for the analysis of helix structure and deformations in oligonucleotides is presented. The parameters "persistence" and "flexibility" as defined in the configurational statistics of polymers of infinite length are reformulated at the oligonucleotide level in an extension of J. A. Schellman's method [(1974) Biopolymers, Vol. 17, pp. 217-226], and used as a basis for a systematic "Persistence Analysis" of the helix deformation properties for all possible subsequences in the structure. The basis for the analysis is a set of link vectors referenced to individual base pairs, and is limited to sequences exhibiting only perturbed rod-like behavior, i.e., below the threshold for supercoiling. The present application of the method is concerned with a physical model for the angular component of bending, so the link vectors are defined as the unit components of a global helix axis obtained by the procedure "Curves" of R. Lavery and H. Sklenar [(1988) J. Biomol. Struct. Dynam., Vol. 6, pp. 63-91; (1989) ibid., Vol. 6, pp. 655-667]. A discussion of the relationship between global bending and relative orientation of base pairs is provided. Our approach is illustrated by analysis of some model oligonucleotide structures with intrinsic kinks, the crystal structure of the dodecamer d(CGCGAATTCGCG)2, and the results of two molecular dynamics simulations on this dodecamer using two variations of the GROMOS force field. The results indicate that essentially all aspects of curvature in short oligonucleotides can be determined, such as the position and orientation of each bend, the sharpness or smoothness, and the location and linearity of subsequences. In the case of molecular dynamics simulations, where a Boltzmann ensemble of structures is analyzed, the spatial extent of the deformations (flexibility) is also considered.


Effects of orally administered fumonisin B₁ (FB₁), partially hydrolysed FB₁, hydrolysed FB₁ and N-(1-deoxy-D-fructos-1-yl) FB₁ on the sphingolipid metabolism in rats.
Fumonisin B1 (FB1) is a Fusarium mycotoxin frequently occurring in maize-based food and feed. Alkaline processing like nixtamalisation of maize generates partially and fully hydrolysed FB1 (pHFB1 and HFB1) and thermal treatment in the presence of reducing sugars leads to formation of N-(1-deoxy-D-fructos-1-yl) fumonisin B1 (NDF). The toxicity of these metabolites, in particular their effect on the sphingolipid metabolism, is either unknown or discussed controversially. We produced high purity FB1, pHFB1a+b, HFB1 and NDF and fed them to male Sprague Dawley rats for three weeks. Once a week, urine and faeces samples were collected over 24 h and analysed for fumonisin metabolites as well as for the sphinganine (Sa) to sphingosine (So) ratio by validated LC-MS/MS based methods. While the latter was significantly increased in the FB1 positive control group, the Sa/So ratios of the partially and fully hydrolysed fumonisins were indifferent from the negative control group. Although NDF was partly cleaved during digestion, the liberated amounts of FB1 did not raise the Sa/So ratio. These results show that the investigated alkaline and thermal processing products of FB1 were, at the tested concentrations, non-toxic for rats, and suggest that according food processing can reduce fumonisin toxicity for humans.


The use of densitometry to detect differences in bone mineral density and content of live White Leghorns fed varying levels of dietary calcium.
Densitometry was investigated as a noninvasive tool to monitor skeletal integrity in live White Leghorns as an indicator for osteoporosis, a noninfectious disease resulting in mineral loss from the bone. The objectives of the experiment were 1) to assess the ability of densitometry to detect differences in bone integrity in live White Leghorns fed varying concentrations of dietary calcium and 2) to correlate densitometric scans with other bone test methods and production parameters that are sensitive to calcium concentrations in the diet. Hens were fed hypercalcemic (5.4%), control (3.6%), or hypocalcemic (1.8%) diets from 32 to 58 wk of age. A Norland densitometer was used to assess bone mineral density (BMD) and bone mineral content (BMC) of the left tibia and humerus in restrained, unanesthetized hens at 36, 46, and 56 wk of age (experiment 1) and at 38, 48, and 58 wk of age (experiment 2). Bones were excised from hens at 38, 48, and 58 wk of age for breaking strength measurements. Results from the densitometric scans showed that BMD and BMC of the humerus and tibia of live hens decreased linearly when hens consumed diets with decreasing concentrations of calcium (experiment 2). Similar trends in BMD and BMC were detected in experiment 1 at 36 wk of age using BW as a covariate. The results from the densitometric scans were comparable to those obtained from other bone tests commonly used. For example, bone breaking force, stress, and modulus of elasticity decreased linearly as hens consumed decreasing concentrations of calcium. Bone breaking force was correlated with BMD (r=0.65, P<0.001). We concluded that densitometry accurately measures differences in BMD and BMC in live birds fed varying concentrations of dietary calcium.


Embryonic skulls of titanosaur sauropod dinosaurs.
Little is known about the cranial anatomy of the taxonomically diverse and geographically widespread titanosaurs, a paucity that has hindered inferences about the genealogical history and evolutionary development of the latest sauropod dinosaurs. Newly discovered fossil eggs containing embryonic remains from the Late Cretaceous of Argentina provide the first articulated skulls of titanosaur dinosaurs. The nearly complete fetal skulls shed light on the evolution of some of the most notable cranial features of sauropod dinosaurs, including the retraction of the external nares, the forward rotation of the braincase, and the abbreviation of the infraorbital region.


Chemoselectivity and the Chan-Lam Coupling Reaction: Adding Amino Acids to Polymer-Coated Microelectrode Arrays.
The placement of a peptide onto a microelectrode array is frequently complicated by the presence of multiple nucleophiles in the peptide. In the work reported here, we have found that the Chan-Lam coupling reactions used to site-selectively place thiol, alcohol, and amine nucleophiles onto diblock-copolymer-coated surfaces are chemoselective for the placement of thiol and alcohol nucleophiles on the arrays. This means that cysteine- and serine-containing peptides can be placed onto an array without any need to protect the N terminus of the peptide. Furthermore, it was found that placement of thiol groups onto an array with the Chan-Lam reaction using optimized reaction times leads to complete coverage of the electrodes. The extent of this coverage can be controlled by varying the reaction time in a manner that allows for the construction of arrays with a gradient of peptide concentrations.


Detection of minimal residual cells carrying the t(14;18) by DNA sequence amplification.
By means of the polymerase chain reaction (PCR) technique, DNA sequences were amplified that flank the crossover sites of a characteristic chromosomal translocation for follicular lymphomas, t(14;18)(q32;q21). This technique permitted the detection of cells carrying the t(14;18) hybrid DNA sequences at a dilution of 1:100,000. The remission marrow and blood samples of a patient with follicular lymphoma and the t(14;18) failed to show any abnormality by morphological examination and conventional Southern blot analysis. However, the t(14;18) hybrid DNA sequences were detected by the PCR technique. Thus, this technique is a highly sensitive tool to detect minimal residual cells carrying the t(14;18) and has the potential to identify a subpopulation of patients with subclinical disease.


Analysis and suppression of parasitic processes in noncollinear optical parametric amplifiers.
The influence of parasitic processes on the performance of ultra-broadband noncollinear optical parametric amplifiers (NOPA's) is investigated for walk-off and non-walk-off compensating configurations. Experimental results with a white-light-seeded NOPA agree well with numerical simulations. The same model shows that 10% of the output energy of an amplified signal can be transferred into a parasitic second harmonic of the signal. These findings are supported by quantitative measurements on a few-cycle NOPA, where a few percent of the signal energy is converted to its second harmonic in the walk-off compensating case. This effect is reduced by an order of magnitude in the non-walk-off compensating configuration. A detailed study of the phase-matching conditions of the most common nonlinear crystals provides guidelines for designing NOPA systems.


Soilborne wheat mosaic virus movement protein and RNA and wheat spindle streak mosaic virus coat protein accumulate inside resting spores of their vector, Polymyxa graminis.
To study virus-vector interactions between Soilborne wheat mosaic virus (SBWMV) or Wheat spindle streak mosaic virus (WSSMV) and Polymyxa graminis Ledingham, P. graminis was propagated in plants grown hydroponically. P. graminis accumulated to high levels in several barley cultivars tested. Multiple developmental stages of P. graminis could be identified in infected barley roots. Accumulation of SBWMV and WSSMV inside P. graminis sporosori in the roots of soil-grown winter wheat and hydroponically grown barley was compared to determine if data obtained from plants naturally infected plants and plants infected by manual inoculation were similar. WSSMV coat protein (CP), SBWMV RNAs, SBWMV movement protein but not SBWMV CP were detected in both soil-grown winter wheat and hydroponically grown barley roots. These data are the first direct evidence that SBWMV and WSSMV are internalized by P. graminis.


Overexpression of SIS2, which contains an extremely acidic region, increases the expression of SWI4, CLN1 and CLN2 in sit4 mutants.
The Saccharomyces cerevisiae SIS2 gene was identified by its ability, when present on a high copy number plasmid, to increase dramatically the growth rate of sit4 mutants. SIT4 encodes a type 2A-related protein phosphatase that is required in late G1 for normal G1 cyclin expression and for bud initiation. Overexpression of SIS2, which contains an extremely acidic carboxyl terminal region, stimulated the rate of CLN1, CLN2, SWI4 and CLB5 expression in sit4 mutants. Also, overexpression of SIS2 in a CLN1 cln2 cln3 strain stimulated the growth rate and the rate of CLN1 and CLB5 RNA accumulation during late G1. The SIS2 protein fractionated with nuclei and was released from the nuclear fraction by treatment with either DNase I or micrococcal nuclease, but not by RNase A. This result, combined with the finding that overexpression of SIS2 is extremely to a strain containing lower than normal levels of histones H2A and H2B, suggests that SIS2 might function to stimulate transcription via an interaction with chromatin.


Percutaneous lateral ankle stabilization: an anatomical investigation.
The current study investigated the topographic anatomy of the percutaneous anatomical lateral ankle stabilization in relation to the neurovascular hindfoot structures. The study should serve as an aid for performance of this new minimally invasive technique. Eleven cadaver specimens were dissected exposing the nerves, vessels, ligaments and tendons. The portals and transosseous tunnels were performed with Kirschner wires. All distances of the Kirschner wires and the neurovascular structures were measured with reference to clearly identifiable bony landmarks. On the medial side the average distance of the Kirschner wire to the medial calcaneal branch of the tibial nerve was found to be 7 mm (SD±4). The medial calcaneal branch was hit twice by the transosseous Kirschner wire. On the lateral side the mean distance of the fibular exit point of the Kirschner wire to the sural nerve was 13 mm (SD±4). The closest distance of the superficial peroneal nerve to the footprint of the ATFL averaged 11.5 mm (SD±3) and its anterior location in relation to the tip of the fibula was 28 mm (SD±8). The posterior tibial artery was found at a mean distance of 41 mm from the perforating Kirschner wire (SD±6). The current study introduced a novel percutaneous approach to treat chronic ankle instability that had minimal risk to neurovascular structures in a cadaver model. Further clinical studies must be undertaken to identify whether this technique would also be superior when compared with open surgery.


Trial of self-expandable metallic stents in the palliation of tracheo-oesophageal fistula in carcinoma of the oesophagus.
Fifty-eight patients with cancer of the oesophagus were studied from July 2002 to November 2005. Thirty-seven patients presented with strictures and 21 presented with tracheo-oesophageal fistula (TOF). There were 35 males and 23 females. Mean age was 57 years. Stents were positioned under fluoroscopic guidance, in the majority of cases under conscious sedation. A contrast study was done on day 1 to assess stent expansion and sealing of the TOF. Data were analysed using Stata Statistcal Software, Release 8.0. Survival was calculated using Kaplan-Meier methodology and log-rank tests were used to assess differences. A p-value < 0.05 was considered to be statistically significant. Stenting was successful in 57 of the 58 patients; 1 perforation occurred at the time of insertion. In total, 68 stents were inserted in 58 patients. Re-stenting was necessary in 10 patients, because of tumour overgrowth (N = 5), stent migration (N = 3) and recurrent fistula (N = 2). Dysphagia improved from a mean score of 2.98 to 1.08. All lesions were squamous cell carcinoma. The positions of the fistulas were proximal (N = 4), middle (N = 14) and distal (N = 3). Over half of the patients in the TOF group had concomitant pneumonia at presentation with decreased survival (p = 0.010) and a hazard ratio of 10.86. Two patients died, on days 4 and 7 respectively. Median survival was 91 days (range 0-273 days) for the stricture group and 62 days (range 3-413 days) for the TOF group, but these differences were not significant (p = 0.945). Covered self-expandable metallic stents provide an acceptable option for the palliation of TOF due to cancer of the oesophagus.


Origami Metamaterials for Tunable Thermal Expansion.
Materials with engineered thermal expansion, capable of achieving targeted area/volume changes in response to variations in temperature, are important for a number of aerospace, optical, energy, and microelectronic applications. While most of the proposed structures with engineered coefficient of thermal expansion consist of bi-material 2D or 3D lattices, here it is shown that origami metamaterials also provide a platform for the design of systems with a wide range of thermal expansion coefficients. Experiments and simulations are combined to demonstrate that by tuning the geometrical parameters of the origami structure and the arrangement of plates and creases, an extremely broad range of thermal expansion coefficients can be obtained. Differently from all previously reported systems, the proposed structure is tunable in situ and nonporous.


Identification and functional characterization of a novel R621C mutation in the synphilin-1 gene in Parkinson's disease.
Synphilin-1 is linked to the pathogenesis of Parkinson's disease (PD) based on its identification as an alpha-synuclein (PARK1) and parkin (PARK2) interacting protein. Moreover, synphilin-1 is a component of Lewy bodies (LB) in brains of sporadic PD patients. Therefore, we performed a detailed mutation analysis of the synphilin-1 gene in 328 German familial and sporadic PD patients. In two apparently sporadic PD patients we deciphered a novel C to T transition in position 1861 of the coding sequence leading to an amino acid substitution from arginine to cysteine in position 621 (R621C). This mutation was absent in a total of 702 chromosomes of healthy German controls. To define a possible role of mutant synphilin-1 in the pathogenesis of PD we performed functional analyses in SH-SY5Y cells. We found synphilin-1 capable of producing cytoplasmic inclusions in transfected cells. Moreover we observed a significantly reduced number of inclusions in cells expressing C621 synphilin-1 compared with cells expressing wild-type (wt) synphilin-1, when subjected to proteasomal inhibition. C621 synphilin-1 transfected cells were more susceptible to staurosporine-induced cell death than cells expressing wt synphilin-1. Our findings argue in favour of a causative role of the R621C mutation in the synphilin-1 gene in PD and suggest that the formation of intracellular inclusions may be beneficial to cells and that a mutation in synphilin-1 that reduces this ability may sensitize neurons to cellular stress.


Improving Rates of Influenza Vaccination Through Electronic Health Record Portal Messages, Interactive Voice Recognition Calls and Patient-Enabled Electronic Health Record Updates: Protocol for a Randomized Controlled Trial.
Clinical decision support (CDS), including computerized reminders for providers and patients, can improve health outcomes. CDS promoting influenza vaccination, delivered directly to patients via an electronic health record (EHR) patient portal and interactive voice recognition (IVR) calls, offers an innovative approach to improving patient care. To test the effectiveness of an EHR patient portal and IVR outreach to improve rates of influenza vaccination in a large multispecialty group practice in central Massachusetts. We describe a nonblinded, randomized controlled trial of EHR patient portal messages and IVR calls designed to promote influenza vaccination. In our preparatory phase, we conducted qualitative interviews with patients, providers, and staff to inform development of EHR portal messages with embedded questionnaires and IVR call scripts. We also provided practice-wide education on influenza vaccines to all physicians and staff members, including information on existing vaccine-specific EHR CDS. Outreach will target adult patients who remain unvaccinated for more than 2 months after the start of the influenza season. Using computer-generated randomization and a factorial design, we will assign 20,000 patients who are active users of electronic patient portals to one of the 4 study arms: (1) receipt of a portal message promoting influenza vaccines and offering online appointment scheduling; (2) receipt of an IVR call with similar content but without appointment facilitation; (3) both (1) and (2); or (4) neither (1) nor (2) (usual care). We will randomize patients without electronic portals (10,000 patients) to (1) receipt of IVR call or (2) usual care. Both portal messages and IVR calls promote influenza vaccine completion. Our primary outcome is percentage of eligible patients with influenza vaccines administered at our group practice during the 2014-15 influenza season. Both outreach methods also solicit patient self-report on influenza vaccinations completed outside the clinic or on barriers to influenza vaccination. Self-reported data from both outreach modes will be uploaded into the EHR to increase accuracy of existing provider-directed EHR CDS (vaccine alerts). With our proposed sample size and using a factorial design, power calculations using baseline vaccination rate estimates indicated that 4286 participants per arm would give 80% power to detect a 3% improvement in influenza vaccination rates between groups (α=.05; 2-sided). Intention-to-treat unadjusted chi-square analyses will be performed to assess the impact of portal messages, either alone or in combination with the IVR call, on influenza vaccination rates. The project was funded in January 2014. Patient enrollment for the project described here completed in December 2014. Data analysis is currently under way and first results are expected to be submitted for publication in 2016. If successful, this study's intervention may be adapted by other large health care organizations to increase vaccination rates among their eligible patients. ClinicalTrials.gov NCT02266277; https://clinicaltrials.gov/ct2/show/NCT02266277 (Archived by WebCite at http://www.webcitation.org/6fbLviHLH).


Effects of a one-year physical activity programme for women with systemic lupus erythematosus - a randomized controlled study.
To study the effects of a one-year physical activity programme on aerobic capacity, physical activity and health-related quality of life (HRQL) in patients with systemic lupus erythematosus (SLE) by a randomized control design. Thirty-five women with low or moderate disease activity and organ damage were randomized to intervention (I) or control (C) group. The intervention during months 0-3 consisted of education, supervised aerobic exercise at high intensity and individual coaching, as well as self-managed physical activity at low-to-moderate intensity. During months 4-12, the physical activity was self-managed and the coaching was successively reduced over time. Outcome measures included: maximal oxygen uptake (VO2 max) from a bicycle ergometer test, self-reported physical activity and HRQL (SF-36). VO2 at sub-max. and max. increased, independent of group, during the one-year study period (main effect of time p < 0.0001). VO2 max. increased between baseline and month 3 (p < 0.0001), between months 3 and 6 (p = 0.01) and the increase was sustained at month 12 (ns). Frequency of physical activity at high intensity also increased, independent of group, during the study period. It was increased at months 3, 6 and 12 compared to baseline (p = 0.02, p < 0.001, p = 0.03). Improvement in mental health between baseline and month 6 (p = 0.002) was seen for the I-group, not the C-group (p = 0.03). Disease activity and organ damage did not change. Physical activity and aerobic capacity increased after supervised exercise and coaching, and the improvement was sustained during the one-year programme. However, no interactions between the group differences were seen, which suggests that repeated measurements could motivate to increased physical activity and thereby to increased aerobic capacity. As sub-max. VO2 increased over time, training-induced changes in VO2 on-kinetics could be another explanation. Little influence on HRQL was seen after the programme. The study indicates that physical activity at high intensity over one year is tolerated by patients with mild to moderate SLE.


Prenatal environmental tobacco smoke exposure increases allergic asthma risk with methylation changes in mice.
Allergic asthma remains an inadequately understood disease. In utero exposure to environmental tobacco smoke (ETS) has been identified as an environmental exposure that can increase an individual's asthma risk. To improve our understanding of asthma onset and development, we examined the effect of in utero ETS exposure on allergic disease susceptibility in an asthmatic phenotype using a house dust mite (HDM) allergen-induced murine model. Pregnant C57BL/6 mice were exposed to either filtered air or ETS during gestation, and their offspring were further exposed to HDM at 6-7 weeks old to induce allergic inflammation. Methylation in the promoter regions of allergic inflammation-related genes and genomic DNA was quantified. Exposure to HDM resulted in the onset of allergic lung inflammation, with an increased presence of inflammatory cells, Th2 cytokines (IL-4, IL-5, and IL-13), and airway remodeling. These asthmatic phenotypes were significantly enhanced when the mice had been exposed to in utero ETS. Furthermore, prenatal ETS exposure and subsequent HDM (ETS/HDM)-induced asthmatic phenotypes agree with methylation changes in the selected asthma-related genes, including IL-4, IL-5, IL-13, INF-γ, and FOXP3. Global DNA methylation was significantly lower in ETS/HDM-exposed mice than that of controls, which coincides with the results observed in lung, spleen, and blood DNAs. Prenatal ETS exposure resulted in a severe increase in allergic inflammatory responses after an HDM challenge, with corresponding methylation changes. Prenatal ETS exposure may influence developmental plasticity and result in altered epigenetic programming, leading to an increased susceptibility to asthma. Environ. Mol. Mutagen. 58:423-433, 2017. © 2017 Wiley Periodicals, Inc.


Understanding risk communication through patient narratives about complex antithrombotic therapies.
The purpose of this study was to explore how patients use narratives to create coherent understandings of risks associated with complex antithrombotic therapies. We led four focus groups consisting of patients older than 65 years of age who had a diagnosis of cardiovascular disease and were using a prescription for cardioprotective agents, such as aspirin, anticoagulants, and/or antiplatelets. The participants' stories were retrospective accounts about physician and patient interactions and adverse events organized in the plot structure of a trial-and-error story. The trial-and-error narrative structure emphasizes patients' idiosyncrasies and reasons why they expect to experience adverse events from changes in treatment. Any fears that they might have had about these risks were mitigated by physician expertise, patient responsibility, and medical technology. Patients who expressed concern about not having sufficient access to medical expertise (e.g., physicians, laboratory tests) seemed less willing to accept risks. The trial-and-error risk narratives helped patients deal with ambiguity and uncertainty about the outcomes of their therapies, and revealed patients' orientations to the risks they faced.


Differential Toll-Like Receptor-Signalling of Burkholderia pseudomallei Lipopolysaccharide in Murine and Human Models.
The Gram-negative bacterium Burkholderia pseudomallei causes melioidosis and is a CDC category B bioterrorism agent. Toll-like receptor (TLR)-2 impairs host defense during pulmonary B.pseudomallei infection while TLR4 only has limited impact. We investigated the role of TLRs in B.pseudomallei-lipopolysaccharide (LPS) induced inflammation. Purified B.pseudomallei-LPS activated only TLR2-transfected-HEK-cells during short stimulation but both HEK-TLR2 and HEK-TLR4-cells after 24 h. In human blood, an additive effect of TLR2 on TLR4-mediated signalling induced by B.pseudomallei-LPS was observed. In contrast, murine peritoneal macrophages recognized B.pseudomallei-LPS solely through TLR4. Intranasal inoculation of B.pseudomallei-LPS showed that both TLR4-knockout(-/-) and TLR2x4-/-, but not TLR2-/- mice, displayed diminished cytokine responses and neutrophil influx compared to wild-type controls. These data suggest that B.pseudomallei-LPS signalling occurs solely through murine TLR4, while in human models TLR2 plays an additional role, highlighting important differences between specificity of human and murine models that may have important consequences for B.pseudomallei-LPS sensing by TLRs and subsequent susceptibility to melioidosis.


Niche marketing production practices for beef cattle in the United States and prevalence of foodborne pathogens.
Niche-marketed food products are rapidly gaining market share in today's society. Consumers are willing to pay premium prices for food perceived to be safer, healthier, more nutritious, and better tasting than conventional food. This review outlines typical production practices for niche-market beef production systems in the United States and compares prevalence estimates of foodborne pathogens in animals and produce from conventional and niche-market production systems. The two main niches for food animal production are organic and natural productions. Organic and natural beef productions are becoming increasingly popular and there is high consumer demand. Two major differences between conventional beef production systems and niche-market production systems (natural and organic) are in the use of antimicrobials and growth-promoting hormones. The impacts of these production systems on foodborne pathogens in beef cattle are variable and often data are nonexistent. Studies directly comparing conventional and niche-market production systems for dairy, swine, poultry, and produce have observed that the prevalence of foodborne pathogens was seldom statistically different between production systems, but when differences were observed, prevalence was typically greater for the niche-market production systems than the conventional production system. The published literature suggests that the perception of niche-marketed food products being safer and healthier for consumers with regard to foodborne pathogens may not be justified.


Functional and Structural Neuroimaging Correlates of Repetitive Low-Level Blast Exposure in Career Breachers.
Combat military and civilian law enforcement personnel may be exposed to repetitive low-intensity blast events during training and operations. Persons who use explosives to gain entry (i.e., breach) into buildings are known as "breachers" or dynamic entry personnel. Breachers operate under the guidance of established safety protocols, but despite these precautions, breachers who are exposed to low-level blast throughout their careers frequently report performance deficits and symptoms to healthcare providers. Although little is known about the etiology linking blast exposure to clinical symptoms in humans, animal studies demonstrate network-level changes in brain function, alterations in brain morphology, vascular and inflammatory changes, hearing loss, and even alterations in gene expression after repeated blast exposure. To explore whether similar effects occur in humans, we collected a comprehensive data battery from 20 experienced breachers exposed to blast throughout their careers and 14 military and law enforcement controls. This battery included neuropsychological assessments, blood biomarkers, and magnetic resonance imaging measures, including cortical thickness, diffusion tensor imaging of white matter, functional connectivity, and perfusion. To better understand the relationship between repetitive low-level blast exposure and behavioral and imaging differences in humans, we analyzed the data using similarity-driven multi-view linear reconstruction (SiMLR). SiMLR is specifically designed for multiple modality statistical integration using dimensionality-reduction techniques for studies with high-dimensional, yet sparse, data (i.e., low number of subjects and many data per subject). We identify significant group effects in these data spanning brain structure, function, and blood biomarkers.


Testing for course patterns in Crohn's disease using clustering analysis.
Using clustering analysis, we sought to identify groups of patients on the basis of the disease course among a population of 177 patients with Crohn's disease and followed for 3 years or more, starting from the first frank exacerbation of the disease. The first 36 values of a monthly clinical score represented the active variables of the clustering analysis. This method yielded 2 course groups, A and B, of 95 and 82 patients respectively. The unfavorable course in group A was characterized by the persistence of the clinically active disease at 3 years, whereas group B individuals achieved complete clinical remission within 2 years of onset on the average. Among the initially known clinical data which could explain the course, only the incidence of an occlusive syndrome was higher in group B, which showed a more favorable course. Although we applied clustering analysis to a patient sample over a period of only 3 years, our results do suggest the existence or 2 primary course groups within the population of patients with Crohn's disease. It would appear that the disease course cannot be predicted from the clinical parameters present at the time of onset, but rather becomes apparent during the course of the first 2 years.


Coupled whispering gallery mode resonators in the Terahertz frequency range.
We report on coupling of two whispering gallery mode resonators in the Terahertz frequency range. Due to the long wavelength in the millimeter to submillimeter range, the resonators can be macroscopic allowing for accurate size and shape control. This is necessary to couple specific modes of two or more resonators. Sets of polyethylene (PE) and quartz disk resonators are demonstrated, with medium (loaded) quality (Q)-factors of 40-800. Both exhibit coinciding resonance frequency spectra over more than ten times the free spectral range. Loading effects of single resonators are investigated which provide strong Q-factor degradation and red-shifts of the resonances in the 0.2% range. By coupling two resonators of the same size, we observe mode splitting, in very good agreement with our numerical calculations.


Balloon dilatation of the aortic valve after previous surgical valvotomy: immediate and follow up results.
To evaluate the immediate and long-term results of transcatheter balloon dilatation of the aortic valve for restenosis after previous surgical valvotomy. Prospective follow up by clinical examination and cross sectional Doppler echocardiography of all patients fulfilling the above criteria. Tertiary paediatric cardiology referral centre. 22 patients (18 male, 4 female)--median (range) age 157.5 (12-254) months--underwent 25 balloon dilatation procedures at a median of 72 (8-155) months after surgery. The median age at surgical valvotomy was 82.5 (0.5-230) months and the systolic gradient across the aortic valve immediately after surgery was 31 (0-49) mm Hg. The indication for dilatation was a Doppler derived peak instantaneous gradient of > 60 mm Hg with grade 2 or less aortic regurgitation. A single balloon was used, and the median balloon to annulus ratio was 1 (0.9-1). After dilatation the catheter pullback gradient decreased acutely from 55 (35-75) to 30 (0-75) mm Hg (p < 0.01) and the Doppler gradient from 74 (52-92) to 40.5 (30-96) mm Hg (p < 0.01). In three patients who underwent a second dilatation of the aortic valve eight months after the first procedure, the pullback gradient decreased from 50 (50-60) to 15 (15-16) mm Hg. Aortic regurgitation grade increased from 1 (0-2) to 2 (1-3); only one patient had grade 3 regurgitation. Over a median follow up of 33 (2-67) months seven patients had aortic valve replacement for recurrent stenosis (six patients) or severe regurgitation (one patient with grade 3 regurgitation after dilatation, who had partial detachment of one of the valve leaflets). There was no significant difference for the pullback gradient (median of 19 v 32.5 mm Hg), Doppler gradient 24 hours after dilatation (33.5 v 50.5 mm Hg; p = 0.03), or the duration of follow up (27.5 v 18 months) between the 12 patients who did not require further dilatation or surgery and the 10 patients who did. Balloon dilatation of the aortic valve is a safe and feasible option for palliation of restenosis after surgical valvotomy for congenital aortic valve stenosis. In many patients, however, stenosis progressed and a further intervention was required.


Indicators of "healthy aging" in older women (65-69 years of age). A data-mining approach based on prediction of long-term survival.
Prediction of long-term survival in healthy adults requires recognition of features that serve as early indicators of successful aging. The aims of this study were to identify predictors of long-term survival in older women and to develop a multivariable model based upon longitudinal data from the Study of Osteoporotic Fractures (SOF). We considered only the youngest subjects (n = 4,097) enrolled in the SOF cohort (65 to 69 years of age) and excluded older SOF subjects more likely to exhibit a "frail" phenotype. A total of 377 phenotypic measures were screened to determine which were of most value for prediction of long-term (19-year) survival. Prognostic capacity of individual predictors, and combinations of predictors, was evaluated using a cross-validation criterion with prediction accuracy assessed according to time-specific AUC statistics. Visual contrast sensitivity score was among the top 5 individual predictors relative to all 377 variables evaluated (mean AUC = 0.570). A 13-variable model with strong predictive performance was generated using a forward search strategy (mean AUC = 0.673). Variables within this model included a measure of physical function, smoking and diabetes status, self-reported health, contrast sensitivity, and functional status indices reflecting cumulative number of daily living impairments (HR >or= 0.879 or RH <or= 1.131; P < 0.001). We evaluated this model and show that it predicts long-term survival among subjects assigned differing causes of death (e.g., cancer, cardiovascular disease; P < 0.01). For an average follow-up time of 20 years, output from the model was associated with multiple outcomes among survivors, such as tests of cognitive function, geriatric depression, number of daily living impairments and grip strength (P < 0.03). The multivariate model we developed characterizes a "healthy aging" phenotype based upon an integration of measures that together reflect multiple dimensions of an aging adult (65-69 years of age). Age-sensitive components of this model may be of value as biomarkers in human studies that evaluate anti-aging interventions. Our methodology could be applied to data from other longitudinal cohorts to generalize these findings, identify additional predictors of long-term survival, and to further develop the "healthy aging" concept.


Skeletal response of male mice to anabolic hormone therapy in the absence of the Igfals gene.
IGF-I is a critical regulator of skeletal acquisition, which acts in endocrine and autocrine/paracrine modes. In serum, IGF-I is carried by the IGF-binding proteins in binary complexes. Further stabilization of these complexes is achieved by binding to the acid labile subunit (ALS) in a ternary complex (of IGF-I-IGF-binding protein 3/5-ALS). Ablation of the Igfals gene in humans (ALS deficiency) and mice (ALS knockout [ALSKO]) leads to markedly decreased serum IGF-I levels, growth retardation, and impaired skeletal acquisition. To investigate whether hormonal replacement therapy would improve the skeletal phenotype in cases of Igfals gene ablation, we treated male ALSKO mice with GH, IGF-I, or a combination of both. Treatments were administered to animals between 4 and 16 weeks of age or from 8 to 16 weeks of age. Although all treatment groups showed an increase (20%) in serum IGF-I levels, there was no increase in body weight, weight gain, or bone length in either age group. Despite the blunted linear growth in response to hormone therapy, ALSKO mice treated with GH showed radial bone growth, which contributed to bone strength tested by 4-point bending. We found that ALSKO mice treated with GH showed increased total cross-sectional area, cortical bone area, and cortical thickness by microtomography. Dynamic histomorphometry showed that although GH and double treatment groups resulted in trends towards increased bone formation parameters, these did not reach significance. However, bone resorption parameters were significantly increased in all treatment groups. ALSKO mice treated between 4 and 16 weeks of age showed minor differences in bone traits compared with vehicle-treated mice. In conclusion, treatment with GH and IGF-I do not work synergistically to rescue the stunted growth found in mice lacking the Igfals gene. Although GH alone appears to increase bone parameters slightly, it does not affect body weight or linear growth.


Studies of the effects of 1,25-dihydroxyvitamin D on skeletal and calcium homeostasis and on inhibition of tumor cell growth.
Vitamin D, parathyroid hormone (PTH), and parathyroid hormone-related peptide (PTHrP) are major regulators of calcium metabolism and vitamin D can also reduce the growth of normal cells and tumor cells. PTHrP and PTH act via a common membrane receptor (PTHR). The mouse PTHR is regulated by a kidney-selective upstream promoter P(1) and ubiquitous downstream promoter P(2). In vitro and in vivo 1,25(OH)(2)D can inhibit PTHR expression in bone but not cartilage by downregulating transcription via P(2). Gene transcription of PTHrP per se can also be downregulated by 1,25(OH)(2)D and by low calcemic vitamin D analogs. This inhibitory effect may reduce the hypercalcemia caused by overproduction of PTHrP by tumor cells. In a malignant keratinoctye cell line, phosphorylation of the retinoid X receptor alpha occurs through the activated Ras-MAP kinase pathway and results in attenuated trans-activation by the vitamin D receptor, its heterodimeric partner. This decreases the growth-inhibitory efficacy of 1,25(OH)(2)D. Studies of the capacity of vitamin D to alter PTHrP production and action and of its anti-proliferative effects can, therefore, shed important light on basic mechanisms controlling these events, and may also have major implications for clinical medicine and therapeutics.


Electronic documentation of central venous catheter-days: validation is essential.
Measurement of central line-associated bloodstream infection (CLABSI) rates outside of intensive care units is challenged by the difficulty in reliably determining central venous catheter (CVC) use. The National Healthcare Safety Network (NHSN) allows for use of electronic data for determination of CVC-days, but validation of electronic data has not been studied systematically. To design and validate a process to reliably measure CVC-days outside of the intensive care units that leverages electronic documentation. Thirty-four inpatient wards at 2 academic hospitals using a common electronic platform for nursing documentation were studied. Electronic queries were created to capture patient and CVC information, and tools and processes for tracking and reporting errors in documentation were developed. Strategies to validate electronic data included comparisons with manual CVC-day determinations and automated data validation using customized tools. Interventions included redesign of documentation interface, real-time audit with feedback of errors, and education. The primary outcome was patient-level total error rate in electronic CVC-day measurement compared with manually counted CVC-days. At baseline, there were a mean (± standard deviation) of [Formula: see text] electronic CVC-day errors (omission and commission errors summed and counted equally) per manually counted CVC-day. After several process improvement cycles over 7 months, the error rate decreased to <0.05 errors per CVC-day and remained at or below this level for 2 years. Baseline electronic CVC-day counts had a high error rate. Stepwise interventions reduced errors to consistently low levels. Validation of electronic calculation of CVC-days is essential to ensure accuracy, particularly if these data will be used for interinstitutional comparison.


Mapping chronic liver disease questionnaire scores onto SF-6D utility values in patients with primary sclerosing cholangitis.
The chronic liver disease questionnaire (CLDQ) is a frequently used liver-specific quality of life instrument, but it does not provide information on preference-adjusted health status, which is essential for cost-utility analysis. We aimed to develop a mapping function deriving utilities from the CLDQ in primary sclerosing cholangitis (PSC). Short form-6D (SF-6D) utilities were calculated from SF-36 data collected in a recent prospective study in which unselected patients with PSC also completed the CLDQ. Ordinary least squares (OLS), generalized linear, median, and kernel regression analyses were employed to devise a mapping function predicting utilities. This was validated in three random subsamples of the cohort and in a separate sample of PSC patients following liver transplantation. Adjusted R (2) and root-mean-square error (RMSE) as well as Pearson's r coefficients and mean absolute errors between predicted and observed values were used to determine model performance. Decompensated liver disease and fatigue, systemic symptoms, and emotional distress, assessed with the CLDQ, were related to worse SF-6D utilities. The final OLS prediction model explained 66.3 % of the variance in the derivation sample. Predicted and observed utilities were strongly correlated (r = 0.807, p < 0.001), but the mean absolute error (0.0604) and adjusted RMSE (10.6 %) were of intermediate size. Similar model characteristics were observed after employment of generalized linear and median regression models and at validation. A model has been constructed, showing good validity predicting SF-6D utilities from CLDQ scores at the group level in PSC. Further testing is required to externally validate the model.


Assessment of proximal femur microarchitecture using ultra-high field MRI at 7 Tesla.
The purpose of this study was to investigate bone microarchitecture of cadaveric proximal femurs using ultra-high field (UHF) 7-Tesla magnetic resonance imaging (MRI) and to compare the corresponding metrics with failure load assessed during mechanical compression test and areal bone mineral density (ABMD) measured using dual-energy X-ray absorptiometry. ABMD of ten proximal femurs from five cadavers (5 women; mean age=86.2±3.8 (SD) years; range: 82.5-90 years) were investigated using dual-energy X-ray absorptiometry and the bone volume fraction, trabecular thickness, trabecular spacing, fractal dimension, Euler characteristics, connectivity density and degree of anisotropy of each femur was quantified using UHF MRI. The whole set of specimens underwent mechanical compression tests to failure. The inter-rater reliability of microarchitecture characterization was assessed with the intraclass correlation coefficient (ICC). Associations were searched using correlation tests and multiple regression analysis. The inter-rater reliability for bone microarchitecture parameters measurement was good with ICC ranging from 0.80 and 0.91. ABMD and the whole set of microarchitecture metrics but connectivity density significantly correlated with failure load. Microarchitecture metrics correlated to each other but did not correlate with ABMD. Multiple regression analysis disclosed that the combination of microarchitecture metrics and ABMD improved the association with failure load. Femur bone microarchitecture metrics quantified using UHF MRI significantly correlated with biomechanical parameters. The multimodal assessment of ABMD and trabecular bone microarchitecture using UHF MRI provides more information about fracture risk of femoral bone and might be of interest for future investigations of patients with undetected osteoporosis.


Photoperiodic stress induces genotype-specific shift in DNA methylation in Tartary buckwheat.
Tartary buckwheat, known for its rich source of health beneficial secondary metabolites, is cultivated in many areas of the world. Among different environmental factors, photoperiod strongly influence its growth, flowering time, and ultimately the yield. In this context, epigenetics could contribute significantly in the regulation of plant response against changing environment. Therefore, with the aim to study the involvement of DNA methylation in photoperiod mediated plant response, genome-wide DNA methylation analysis was performed in two accessions (A1 and A2) of Tartary buckwheat using three photoperiodic treatments, i.e., 10-hr light/day (T1), 12-hr light/day (T2), and 14-hr light/day (T3). Flowering time and plant fresh weight data revealed that accessions A1 and A2 prefer T1 and T2 treatments, respectively. Total DNA methylation ratio increased with the increase in photoperiod in accession A1 but decreased under same conditions in accession A2. Full methylation increased significantly while intensive decrease in hemimethylation was noted from T2 to T3 in A1, whereas full methylation strongly increased and hemimethylation strongly decreased from T1 to T2 in A2. Overall, the DNA methylation events appeared more frequently than demethylation events. This study reports for the first time an accession-/ genotype specific pattern of shift in the DNA methylation under different photoperiodic treatments that will pave the way toward identification of specific genes involved in the regulation of plant response against photoperiodic stress.


Comparison of the efficiency of rat papillary muscles during afterloaded isotonic contractions and contractions with sinusoidal length changes.
The results of previous studies suggest that the maximum mechanical efficiency of rat papillary muscles is lower during a contraction protocol involving sinusoidal length changes than during one involving afterloaded isotonic contractions. The aim of this study was to compare directly the efficiency of isolated rat papillary muscle preparations in isotonic and sinusoidal contraction protocols. Experiments were performed in vitro (27 degrees C) using left ventricular papillary muscles from adult rats. Each preparation performed three contraction protocols: (i) low-frequency afterloaded isotonic contractions (10 twitches at 0.2 Hz), (ii) sinusoidal length change contractions with phasic stimulation (40 twitches at 2 Hz) and (iii) high-frequency afterloaded isotonic contractions (40 twitches at 2 Hz). The first two protocols resembled those used in previous studies and the third combined the characteristics of the first two. The parameters for each protocol were adjusted to those that gave maximum efficiency. For the afterloaded isotonic protocols, the afterload was set to 0.3 of the maximum developed force. The sinusoidal length change protocol incorporated a cycle amplitude of +/-5% resting length and a stimulus phase of -10 degrees. Measurements of force output, muscle length change and muscle temperature change were used to calculate the work and heat produced during and after each protocol. Net mechanical efficiency was defined as the proportion of the energy (enthalpy) liberated by the muscle that appeared as work. The efficiency in the low-frequency, isotonic contraction protocol was 21.1+/-1.4% (mean +/- s.e.m., N=6) and that in the sinusoidal protocol was 13.2+/-0.7%, consistent with previous results. This difference was not due to the higher frequency or greater number of twitches because efficiency in the high-frequency, isotonic protocol was 21.5+/-1.0%. Although these results apparently confirm that efficiency is protocol-dependent, additional experiments designed to measure work output unambiguously indicated that the method used to calculate work output in isotonic contractions overestimated actual work output. When net work output, which excludes work done by parallel elastic elements, rather than total work output was used to determine efficiency in afterloaded isotonic contractions, efficiency was similar to that for sinusoidal contractions. The maximum net mechanical efficiency of rat papillary muscles performing afterloaded isotonic or sinusoidal length change contractions was between 10 and 15%.


Sequence polymorphism and expression variability of Crassostrea gigas immune related genes discriminate two oyster lines contrasted in term of resistance to summer mortalities.
Summer mortalities of Crassostreagigas are a major concern in oyster aquaculture. They are the result of a complex interaction between the host, pathogens and environmental factors. Oyster genetics have been identified as an essential determinant of oyster susceptibility to summer mortalities. As the capability of oysters to circumvent diseases depends in part on their immune defenses, we aimed to analyze the gene expression and sequence polymorphism of 42 immune related genes in two oyster lines selected for their "High" (H) and "Low" (L) survival to summer mortalities. Results showed that the variability of gene expression and the sequence polymorphism acting on particular genes could enable the discrimination between H and L oyster lines. Besides, a higher sequence polymorphism was observed on the L line affecting 11 of the 42 analyzed genes. By analyzing gene expression, sequence polymorphism and gene copy number of two antimicrobial peptide families (Cg-Defs and Cg-Prp), and an antimicrobial protein (Cg-BPI) on individual oysters, we showed that gene expression and/or sequence polymorphism could also discriminate H and L oyster lines. Finally, we observed a positive correlation between the gene expression and the gene copy number of antimicrobials and that sequence polymorphism could be encoded in the genome. Overall, this study gives new insights in the relationship between oyster immunity and divergent phenotypes, and discusses the potential implication of antimicrobial diversity in oyster survival to summer mortalities.


Central effect of intravenous phenylephrine on baroreflex control of renal nerves.
Responses of renal sympathetic nerve activity were determined in eight chloralose-anesthetized rabbits during sustained (1-3 minutes) increases in arterial pressure induced by phenylephrine infusion, and as arterial pressure returned to control. In four of the eight experiments, aortic baroreceptor traffic was also recorded. When arterial pressure was raised from 81 +/- 5 to 110 +/- 7 mm Hg, renal nerve activity decreased from 30 +/- 7 to 1 +/- 1 imp/sec. Aortic nerve activity increased from 208 +/- 35 to 346 +/- 49 imp/sec. When pressure returned to control (81 +/- 5 mm Hg), renal nerve activity remained inhibited (7 +/- 2 imp/sec), even though aortic nerve activity had also returned to control (195 +/- 33 imp/sec). Arterial pressure and traffic in the renal and aortic nerves returned to control over the succeeding 1 to 5 minutes. Transient increases in arterial pressure (lasting less than 1 minute) due to bolus injections of phenylephrine resulted in inhibition of renal nerve traffic followed by rapid recovery. In five rabbits with aortic and vagal nerves sectioned and both carotid sinuses isolated from the circulation, intravenous phenylephrine infusion augmented the gain of the isolated carotid baroreflex (particularly at low carotid sinus pressures). In nine experiments, injection of phenylephrine (0.01, 0.1, or 1.0 microgram) into the lateral ventricles did not change the basal renal nerve traffic but augmented the gain of the baroreflex control of the renal nerves. Our data indicate that peripherally infused phenylephrine can alter the arterial baroreflex control of the renal nerves by a central effect. The similar influence of intracerebroventricular phenylephrine on baroreflex control of the renal nerves is consistent with this view.


Microstructure of subretinal drusenoid deposits revealed by adaptive optics imaging.
Subretinal drusenoid deposits (SDD), a recently recognized lesion associated with progression of age-related macular degeneration, were imaged with adaptive optics scanning laser ophthalmoscopy (AO-SLO) and optical coherence tomography (AO-OCT). AO-SLO revealed a distinct en face structure of stage 3 SDD, showing a hyporeflective annulus surrounded reflective core packed with hyperreflective dots bearing a superficial similarity to the photoreceptors in the unaffected retina. However, AO-OCT suggested that the speckled appearance over the SDD rendered by AO-SLO was the lesion material itself, rather than photoreceptors. AO-OCT assists proper interpretation and understanding of the SDD structure and the lesions' impact on surrounding photoreceptors produced by AO-SLO and vice versa.


Modulatory role of GSTM1 null genotype on the frequency of micronuclei in pesticide-exposed agricultural workers.
In this study, we aimed to investigate the extent of genotoxic risk and the association between null GSTM1/GSTT1 and GSTP1 Ile105Val variants and cellular DNA damage, as measured by micronucleus (MN) assay in a group of agricultural workers from Denizli, Turkey. Peripheral blood samples were collected from 116 subjects, including 58 workers who were occupationally exposed to pesticides and 58 healthy unexposed controls. The MN frequencies of each individual were assessed by cytokinesis-blocked micronuclei assays on lymphocytes. Genotypes for different GST variants were determined using polymerase chain reaction-based methods. A significant 3.4-fold increase in MN frequency was observed in workers compared with the controls (p < 0.001). Among the GST genotypes, only the GSTM1 null genotype was found to be significantly associated with an increased MN frequency in workers (p = 0.01). Individuals with a concomitant null GSTM1/GSTT1 genotype demonstrated a significant (p = 0.01) increase in MN frequency compared with those with functional isozymes in the exposed worker group. The association of the GSTM1 null genotype with higher MN frequency suggests that it may be a modifier of genotoxic risk in individuals exposed to pesticides and may thus be a candidate susceptibility biomarker for human biomonitoring studies.


Influence of light-curing units and restorative materials on the micro hardness of resin cements.
The aim of this study was to evaluate the effect of indirect restorative materials (IRMs) and light-curing units (LCUs) on the micro hardness of dual-cured resin cement. A total of 36 cylindrical samples (2 mm thick) were prepared with dual-cured resin cement (Relyx ARC) photo-activated with either a QTH (Optilight Plus) for 40s or a LED (Radii) light-curing unit for 65s. Photo-activation was performed through the 2-mm- thick IRMs and the samples were divided into six groups (n=6) according to the combination of veneering materials (without, ceramic and indirect resin) and LCUs (QTH and LED). In the control group, the samples were light-cured with a QTH unit without the interposition of any restorative material. Vickers micro hardness test was performed on the top and bottom surfaces of each sample (load of 50 g for 15 secs). The data were statistically analyzed using a three-way ANOVA followed by Tukey x s post-hoc test ( P < 0.05). There were no statistically significant differences on the top surface between the light curing-units ( P > 0.05); however, the LED provided greater hardness on the bottom surface when a ceramic material was used ( P < 0.05). The mean hardness in photo-activated samples, in which there was no interposition of indirect materials, was significantly greater ( P < 0.01). It may be concluded that the interposition of the restorative material decreased the micro hardness in the deeper cement layer. Such decrease, however, was lower when the ceramic was interposed and the cement light-cured with LED.


Hybrid biological, electron beam and zero-valent nano iron treatment of recalcitrant metalworking fluids.
Hybrid approaches for the remediation and detoxification of toxic recalcitrant industrial wastewater were investigated. The focus was waste metalworking fluid, which was selected as a representative model of other waste streams that are toxic, recalcitrant and that require more sustainable routes of safe disposal. The hybrid approaches included biodegradation, electron beam irradiation and zero-valent nano iron advanced oxidation processes that were employed individually and in sequence employing a factorial design. To compare process performance operationally exhausted and pristine metalworking fluid were compared. Sequential hybrid electron beam irradiation, biological, nanoscale zero-valent iron and biological treatment lead to synergistic detoxification and degradation of both recalcitrant streams, as determined by complementary surrogates and lead to overall improved COD removal of 92.8 ± 1.4% up from 85.9 ± 3.4% for the pristine metalworking fluid. Electron beam pre-treatment enabled more effective biotreatment, achieving 69.5 ± 8% (p = 0.005) and 24.6 ± 4.8% (p = 0.044) COD reductions.


Synergy-COPD: a systems approach for understanding and managing chronic diseases.
Chronic diseases (CD) are generating a dramatic societal burden worldwide that is expected to persist over the next decades. The challenges posed by the epidemics of CD have triggered a novel health paradigm with major consequences on the traditional concept of disease and with a profound impact on key aspects of healthcare systems. We hypothesized that the development of a systems approach to understand CD together with the generation of an ecosystem to transfer the acquired knowledge into the novel healthcare scenario may contribute to a cost-effective enhancement of health outcomes. To this end, we designed the Synergy-COPD project wherein the heterogeneity of chronic obstructive pulmonary disease (COPD) was addressed as a use case representative of CD. The current manuscript describes main features of the project design and the strategies put in place for its development, as well the expected outcomes during the project life-span. Moreover, the manuscript serves as introductory and unifying chapter of the different papers associated to the Supplement describing the characteristics, tools and the objectives of Synergy-COPD.