[Internet addiction: development and validation of an instrument in adolescent scholars in Lima, Peru].
To develop and validate an instrument to assess Internet Addiction (IA) phenomenon in adolescents of Metropolitan Lima. We performed an observational analytical study, including a sample of 248 high school adolescent students. In order to evaluate the IA, we constructed the questionnaire: "Scale for Internet Addiction of Lima" (SIAL), which assesses symptoms and dysfunctional characteristics. The resulting items were submitted to experts' judgment, finally obtaining a 11-item scale. The mean age was 14 years old. The psychometric analysis of the instrument showed a Cronbach' Alpha Coefficient of 0.84, with values of item-total correlation ranging from 0.45 to 0.59. The dimensional analysis yielded a two-dimensional structure that explained up to 50.7% of the total variance. The bi-dimensional data analysis revealed a significant association (p<0,001) between Dimension I (symptoms of IA) and the weekly time spent on the Internet, male sex, past history of bad behavior in school and plans for the future. Dimension II (dysfunction due to IA) had a significant association to past history of bad behavior, plans for the future (p<0,001) and missing school without valid reasons. The SIAL showed a good internal consistency, with moderate and significant inter-item correlations. The findings show that addiction has a dynamic role, which evidences a problem generated in family patterns and inadequate social networks.


Parthenogenesis: does it occur spontaneously in mice?
If parthenogenesis occurred in bisexual organisms, it would produce an excess of females and depress the sex ratio. The phenotypes of female mice, from matings that produce an excess of females, were examined for evidence of the presence of marker genes of paternal origin. All proved to be hybrids of the maternal and paternal strains, thus excluding parthenogenesis as the cause of the low sex ratio.


A novel processed food classification system applied to Australian food composition databases.
The extent of food processing can affect the nutritional quality of foodstuffs. Categorising foods by the level of processing emphasises the differences in nutritional quality between foods within the same food group and is likely useful for determining dietary processed food consumption. The present study aimed to categorise foods within Australian food composition databases according to the level of food processing using a processed food classification system, as well as assess the variation in the levels of processing within food groups. A processed foods classification system was applied to food and beverage items contained within Australian Food and Nutrient (AUSNUT) 2007 (n = 3874) and AUSNUT 2011-13 (n = 5740). The proportion of Minimally Processed (MP), Processed Culinary Ingredients (PCI) Processed (P) and Ultra Processed (ULP) by AUSNUT food group and the overall proportion of the four processed food categories across AUSNUT 2007 and AUSNUT 2011-13 were calculated. Across the food composition databases, the overall proportions of foods classified as MP, PCI, P and ULP were 27%, 3%, 26% and 44% for AUSNUT 2007 and 38%, 2%, 24% and 36% for AUSNUT 2011-13. Although there was wide variation in the classifications of food processing within the food groups, approximately one-third of foodstuffs were classified as ULP food items across both the 2007 and 2011-13 AUSNUT databases. This Australian processed food classification system will allow researchers to easily quantify the contribution of processed foods within the Australian food supply to assist in assessing the nutritional quality of the dietary intake of population groups.


Light propagation in a two-component nonlinear composite medium.
The propagation of intense light through a two-component nonlinear heterogeneous medium is considered by treating the medium as a composite medium. Introduction of dispersion and Kerr-type nonlinear terms leads to a nonlinear Schrödinger-type scattering equation. The effect of nonlinear scattering loss on soliton propagation is studied numerically. The propagation of two oppositely traveling waves is also discussed.


Transcriptional regulation of the sodium channel gene (SCN5A) by GATA4 in human heart.
Aberrant expression of the sodium channel gene (SCN5A) has been proposed to disrupt cardiac action potential and cause human cardiac arrhythmias, but the mechanisms of SCN5A gene regulation and dysregulation still remain largely unexplored. To gain insight into the transcriptional regulatory networks of SCN5A, we surveyed the promoter and first intronic regions of the SCN5A gene, predicting the presence of several binding sites for GATA transcription factors (TFs). Consistent with this prediction, chromatin immunoprecipitation (ChIP) and sequential ChIP (Re-ChIP) assays show co-occupancy of cardiac GATA TFs GATA4 and GATA5 on promoter and intron 1 SCN5A regions in fresh-frozen human left ventricle samples. Gene reporter experiments show GATA4 and GATA5 synergism in the activation of the SCN5A promoter, and its dependence on predicted GATA binding sites. GATA4 and GATA6 mRNAs are robustly expressed in fresh-frozen human left ventricle samples as measured by highly sensitive droplet digital PCR (ddPCR). GATA5 mRNA is marginally but still clearly detected in the same samples. Importantly, GATA4 mRNA levels are strongly and positively correlated with SCN5A transcript levels in the human heart. Together, our findings uncover a novel mechanism of GATA TFs in the regulation of the SCN5A gene in human heart tissue. Our studies suggest that GATA5 but especially GATA4 are main contributors to SCN5A gene expression, thus providing a new paradigm of SCN5A expression regulation that may shed new light into the understanding of cardiac disease.


An assessment of critical issues facing the clinical laboratory workforce.
The National Center for Health Workforce Analysis of the Health Resources and Services Administration (HRSA), Bureau of Health Professions contracted with the University of California at San Francisco Center for the Health Professions to conduct a study of the clinical laboratory workforce. The study utilized existing and new survey data, in collaboration with the American Society for Clinical Pathology (ASCP), as well as interviews with experts in the field. This paper will describe the demographic characteristics, roles, and scope of practice of this workforce, as well as identify factors that influence the current and future demand for and supply of workers. There is ample evidence of a shortage of workers over the past few years, indicated by double-digit vacancy rates, increasing salaries, and the use of other incentives to fill positions. Recent data show a reduction in vacancy rates, although an increase in per diem and contract workers may mask a continuing shortage. There is recent growing interest in the profession, as evidenced by an increase of applicants to educational programs. Several factors make it difficult to predict the future need for and role of clinical laboratory workers, including an aging population and the demand for increased services, the possible retirement of many current workers, automation, new technology and tests, and public health challenges.


Overview on urban and peri-urban agriculture: definition, impact on human health, constraints and policy issues.
To collate and synthesize current knowledge of components of urban agriculture (UA) with a thematic emphasis on human health impact and a geographic emphasis on East Africa. Data management followed a structured approach in which key issues were first identified and then studies selected through literature search and personal communication. Evidence-based principles. Urban agriculture is an important source of food security for urban dwellers in East Africa. Descriptors of UA are location, areas, activities, scale, products, destinations, stakeholders and motivation. Many zoonotic and food-borne diseases have been associated with UA but evidence on human health impact and management is lacking. Major constraints to UA are illegality and lack of access to input and market; policy options have been developed for overcoming these. Urban agriculture is an important activity and likely to remain so. Both positive and negative human health impacts are potentially important but more research is needed to understand these and set appropriate policy and support levels.


Treating intrusions, promoting resilience: an overview of therapies for trauma-related psychological disorders.
The efficacy of psychotherapeutic approaches in the treatment of posttraumatic stress disorder (PTSD) can be regarded as empirically demonstrated. Overall, effect sizes appear to be higher for psychotherapy than for medication. Many well-controlled trials with a mixed variety of trauma survivors have demonstrated that trauma-focused cognitive-behavioral therapy (TF-CBT) is effective in treating PTSD. Prolonged exposure therapy (PE) is currently seen as the treatment with the strongest evidence for its efficacy. Cognitive therapy (CT) and cognitive processing therapy (CPT), with their stronger emphasis on cognitive techniques, and Eye Movement Desensitization and Reprocessing (EMDR) seem equally effective. More recent developments include brief eclectic psychotherapy for PTSD (BEPP) and narrative exposure therapy (NET). Emerging evidence shows that TF-CBT can successfully be applied in PTSD patients suffering from severe comorbidities such as borderline personality disorder or substance abuse disorder (Schnyder & Cloitre, 2015). There is also a trend towards developing "mini-interventions," that is, short modules tailored to approach specific problems. Moreover, evidence-based approaches should be complemented by interventions that aim at promoting human resilience to stress. Finally, given the globalization of our societies (Schnyder, 2013), culture-sensitive psychotherapists should try to understand the cultural components of a patient's illness and help-seeking behaviors, as well as their expectations with regard to treatment.


The harm argument against surrogacy revisited: two versions not to forget.
It has been a common claim that surrogacy is morally problematic since it involves harm to the child or the surrogate-the harm argument. Due to a growing body of empirical research, the harm argument has seen a decrease in popularity, as there seems to be little evidence of harmful consequences of surrogacy. In this article, two revised versions of the harm argument are developed. It is argued that the two suggested versions of the harm argument survive the current criticism against the standard harm argument. The first version argues that the child is harmed by being separated from the gestational mother. The second version directs attention to the fact that surrogacy involves great incentives to keep the gestational mother's level of maternal-fetal attachment low, which tend to increase the risk of harm to the child. While neither of the two arguments is conclusive regarding the moral status of surrogacy, both constitute important considerations that are often ignored.


Kinematical, Structural and Mechanical Adaptations to Desiccation in Poikilohydric Ramonda myconi (Gesneriaceae).
Resurrection plants have fascinated scientists since centuries as they can fully recover from cellular water contents below 10%, concomitantly showing remarkable leaf folding motions. While physiological adaptations have been meticulously investigated, the understanding of structural and mechanical adaptations of this phenomenon is scarce. Using imaging and bending techniques during dehydration-rehydration experiments, morphological, anatomical, and biomechanical properties of desiccation-tolerant Ramonda myconi are examined, and selected structural adaptations are compared to those of homoiohydrous Monophyllaea horsfieldii (both Gesneriaceae). At low water availability, intact and cut-off R. myconi leaves undergo considerable morphological alterations, which are fully and repeatedly reversible upon rehydration. Furthermore, their petioles show a triphasic mechanical behavior having a turgor-based structural stability at high (Phase 1), a flexible mechanically state at intermediate (Phase 2) and a material-based stability at low water contents (Phase 3). Lastly, manipulation experiments with cut-off plant parts revealed that both the shape alterations of individual structures, as well as, the general leaf kinematics largely rely on passive swelling and shrinking processes. Taken together, R. myconi possesses structural and mechanical adaptations to desiccation (in addition to physiological adaptations), which may mainly be passively driven by its water status influenced by the water fluctuations in its surroundings.


Risk factors for infection and role of C-reactive protein in Korean patients with systemic lupus erythematosus.
To evaluate risk factors for infection and the role of C-reactive protein (CRP) in the diagnosis of infection, a retrospective case control study was performed among Korean systemic lupus erythematosus patients. Of 120 proven infections, 31 episodes (25.8%) occurred in patients taking no corticosteroids (CS). The risk of infection was lower in patients taking low-dose CS (< 300 mg prednisolone/month) than no CS (odds ratio (OR) 0.36). In patients receiving high-dose CS (> 1000 mg prednisolone/month), however, the risk increased (OR 2.9). In patients taking no CS, disease activity manifested as increased SLEDAI, anemia and active urinary sediment, was associated with infection. The CRP was higher in the patients with infection than controls and the CRP levels over 50 mg/l were observed only in infection. These results suggest that CS have a bimodal influence on infection depending on dose. Disease activity is an important risk factor for infection in patients taking no CS. Finally, CRP levels greater than 50 mg/l suggest the presence of infection.


The role of the extracellular loops of the CGRP receptor, a family B GPCR.
The CGRP (calcitonin gene-related peptide) receptor is a family B GPCR (G-protein-coupled receptor). It consists of a GPCR, CLR (calcitonin receptor-like receptor) and an accessory protein, RAMP1 (receptor activity-modifying protein 1). RAMP1 is needed for CGRP binding and also cell-surface expression of CLR. There have been few systematic studies of the ECLs (extracellular loops) of family B GPCRs. However, they are likely to be especially important for the interaction of the N-termini of the peptide agonists that are the natural agonists for these receptors. We have carried out alanine scans on all three ECLs of CLR, as well as their associated juxtamembrane regions. Residues within all three loops influence CGRP binding and receptor activation. Mutation of Ala203 and Ala206 on ECL1 to leucine increased the affinity of CGRP. Residues at the top of TM (transmembrane) helices 2 and 3 influenced CGRP binding and receptor activation. L351A and E357A in TM6/ECL3 reduced receptor expression and may be needed for CLR association with RAMP1. ECL2 seems especially important for CLR function; of the 16 residues so far examined in this loop, eight residues reduce the potency of CGRP at stimulating cAMP production when mutated to alanine.


Responses of the HPA axis after chronic variable stress: effects of novel and familiar stressors.
We examined the role that novelty plays in determining interactions between chronic and acute stress, when both the chronic and acute stressors emphasize emotional processing (i.e. stressful stimuli that do not present immediate threats to somatic homeostasis, and are processed primarily by limbic and forebrain circuits). Rats were exposed to a chronic variable stress (CVS) regimen, and were subsequently tested to evaluate responses to novel and familiar acute stressors. One group was exposed to CVS that included restraint, and was then tested with this familiar stressor. Another group was exposed to CVS that did not include restraint, and were tested with restraint as a novel stressor. Additional rats were not chronically stressed. Plasma adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) were assayed. When the rats were exposed to familiar acute stress after CVS, ACTH responses were blunted. The ACTH responses were normal in the rats that were tested with novel acute stress--the responses resembled those of rats that had no prior stress experience. CORT responses did not differ between the groups, regardless of stress history. Despite the fact that all the chronic and acute stressors emphasized emotional processing of aversive stimuli, and thus likely involved overlapping limbic and forebrain circuits, the hormonal responses differed depending upon familiarity with the acute stressor. Further research is required to identify the neuronal mechanisms that mediate these differing responses to novel and familiar emotional stressors.


Honokiol for cancer therapeutics: A traditional medicine that can modulate multiple oncogenic targets.
In spite of billions of dollars expended on cancer research every year, the incidence rate and the mortality rate due to this widespread disease has increased drastically over the last few decades. Recent reports from the World Health Organization advocate that overall global cancer burden and deaths due to cancer are expected to double by the next decade. Synthetic drugs developed as chemotherapeutics have repeatedly shown adverse side effects and development of chemoresistance. Cancer is basically a multifactorial disease that necessitates the modulation of multiple targets and oncogenic signaling pathways. Honokiol (C18H18O2) is a biphenolic natural compound isolated from the leaves and barks of Magnolia plant species and has been extensively studied for its beneficial effects against several chronic diseases. Honokiol is capable of efficiently preventing the growth of wide variety of tumors such as those of brain, breast, cervical, colon, liver, lung, prostate, skin, and hematological malignancies. Recent work has shown that this phytochemical can modulate various molecular targets such as activation of pro-apoptotic factors, suppression of anti-apoptotic proteins and different transcription factors, downregulation of various enzymes, chemokines, cell surface adhesion molecules, and cell cycle proteins, and inhibition of activity of protein tyrosine kinases and serine/threonine kinases. Because of its pharmacological safety, honokiol can either be used alone or in combination with other chemotherapeutic drugs for the prevention and treatment of cancer. The current review describes in detail the various reports supporting these anti-cancer studies documented with this promising agent.


FBH1 helicase disrupts RAD51 filaments in vitro and modulates homologous recombination in mammalian cells.
Efficient repair of DNA double strand breaks and interstrand cross-links requires the homologous recombination (HR) pathway, a potentially error-free process that utilizes a homologous sequence as a repair template. A key player in HR is RAD51, the eukaryotic ortholog of bacterial RecA protein. RAD51 can polymerize on DNA to form a nucleoprotein filament that facilitates both the search for the homologous DNA sequences and the subsequent DNA strand invasion required to initiate HR. Because of its pivotal role in HR, RAD51 is subject to numerous positive and negative regulatory influences. Using a combination of molecular genetic, biochemical, and single-molecule biophysical techniques, we provide mechanistic insight into the mode of action of the FBH1 helicase as a regulator of RAD51-dependent HR in mammalian cells. We show that FBH1 binds directly to RAD51 and is able to disrupt RAD51 filaments on DNA through its ssDNA translocase function. Consistent with this, a mutant mouse embryonic stem cell line with a deletion in the FBH1 helicase domain fails to limit RAD51 chromatin association and shows hyper-recombination. Our data are consistent with FBH1 restraining RAD51 DNA binding under unperturbed growth conditions to prevent unwanted or unscheduled DNA recombination.


Role of Organic Ligands Orientation on the Geometrical and Optical Properties of Au25(SCH3)180.
The role of the organic group orientation on the geometrical and optical properties in a neutral Au25 nanocluster has been analyzed through density functional theory (DFT) and time-dependent density functional theory (TDDFT) simulations. Starting from two different X-ray diffraction (XRD) resolved structures which differ in the ligand orientation, we optimized the methyl substituted neutral nanoclusters at the B3LYP//6-31G (d,p)/LANL2DZ level, finding remarkable differences on the bond length and the symmetry of the gold kernels. Despite these differences, the TDDFT estimated absorption features of the two clusters are quite similar, showing that ligand orientation brings minor effects on the optical properties of the nanoclusters. All obtained results are in good agreement with available experimental data.


How Does Mindfulness Meditation Work? Proposing Mechanisms of Action From a Conceptual and Neural Perspective.
Cultivation of mindfulness, the nonjudgmental awareness of experiences in the present moment, produces beneficial effects on well-being and ameliorates psychiatric and stress-related symptoms. Mindfulness meditation has therefore increasingly been incorporated into psychotherapeutic interventions. Although the number of publications in the field has sharply increased over the last two decades, there is a paucity of theoretical reviews that integrate the existing literature into a comprehensive theoretical framework. In this article, we explore several components through which mindfulness meditation exerts its effects: (a) attention regulation, (b) body awareness, (c) emotion regulation (including reappraisal and exposure, extinction, and reconsolidation), and (d) change in perspective on the self. Recent empirical research, including practitioners' self-reports and experimental data, provides evidence supporting these mechanisms. Functional and structural neuroimaging studies have begun to explore the neuroscientific processes underlying these components. Evidence suggests that mindfulness practice is associated with neuroplastic changes in the anterior cingulate cortex, insula, temporo-parietal junction, fronto-limbic network, and default mode network structures. The authors suggest that the mechanisms described here work synergistically, establishing a process of enhanced self-regulation. Differentiating between these components seems useful to guide future basic research and to specifically target areas of development in the treatment of psychological disorders.


Prevalence of erythrocyte haemoglobin H inclusions in unselected patients with clonal myeloid disorders.
Patients with clonal myeloid disorders, especially myelodysplastic syndromes (MDS), may acquire alpha-thalassaemia. To estimate the prevalence of this erythrocyte phenotype, we examined brilliant cresyl blue-stained blood smears from 201 patients with neoplastic myeloid disorders and 282 controls (195 non-clonal anaemia, 62 with medical illnesses without anaemia and 25 healthy persons). Haemoglobin H inclusions were detected in 8/100 patients with MDS (8%) and 2/81 (2.5%) patients with myeloproliferative disorders, but in none of the acute leukaemia patients or controls. We conclude that the emergence of thalassaemic clones may be relatively common in the disordered marrow milieu of MDS.


The coccidioidal complement fixation and immunodiffusion-complement fixation antigen is a chitinase.
Culture filtrates and autolysates of Coccidioides immitis have provided suitable crude antigens for the serodiagnosis and prognosis of coccidioidomycosis. One of these, a heat-labile antigen which participates in the immunodiffusion reaction corresponding to the complement fixation reaction (IDCF), has been characterized as a 110-kDa native protein that, when subjected to reducing conditions and heat, yields a 48-kDa component. The present report provides serologic and biochemical evidence that this antigen is a chitinase. This chitinase, isolated from 48-h culture filtrate of the spherule-endospore-phase C. immitis by affinity adsorption to chitin, formed a line of identity with the IDCF reference antigen and participated in the complement fixation reaction with human serum. It lost its enzymatic as well as antigenic activity when heated, but when not heated it retained its enzymatic activity even when precipitated with coccidiodal antibody present in human serum. This chitinase represents a significant serodiagnostic substance and may be important in the morphogenesis of C. immitis.


Identification of a major maturation-activated acetyl-CoA carboxylase kinase in sea star oocytes as p44mpk.
Maturation-activated protein-serine/threonine kinases were investigated in the high-speed supernatant fractions from sea-star oocytes harvested at the time of germinal vesicle breakdown. One of the major stimulated protein kinases able to phosphorylate acetyl-CoA carboxylase in these extracts was found to co-purify with a 44 kDa myelin basic protein kinase (p44mpk) that is activated with a similar time course during oocyte maturation. Purified sea-star oocyte p44mpk phosphorylated acetyl-CoA carboxylase (purified from rat liver) predominantly on serine and to a small extent on threonine. Furthermore, the phosphorylation of acetyl-CoA carboxylase occurred principally on a tryptic phosphopeptide which displayed electrophoretic and chromatographic properties very similar to those of the peptide that has previously been shown to undergo increased phosphorylation in response to insulin in rat adipocytes [Brownsey & Denton (1982) Biochem. J. 202, 77-86]. The acetyl-CoA carboxylase was phosphorylated at a similar rate and to a similar extent by casein kinase II, which was also purified from maturing sea-star oocytes. Although casein kinase II was also activated approximately 3-fold near the time of nuclear envelope breakdown, it was responsible for only a minor component of the total enhanced acetyl-CoA carboxylase kinase activity measured in the soluble extracts from maturing oocytes. Acetyl-CoA carboxylase was a relatively poor substrate for the major S6 peptide kinase activity that was also stimulated during resumption of meiosis in the oocytes. The properties of the p44mpk are reminiscent of those of a microtubule-associated protein 2 (MAP-2) kinase that is activated in response to insulin and other mitogens in mammalian cells [Ray & Sturgill (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 3753-3757; Hoshi, Nishida & Sakai (1988) J. Biol. Chem. 263, 5396-5401]. It is intriguing that several of the mammalian protein kinases that are acutely activated after mitogenic prompting of quiescent mouse fibroblasts (i.e. G0 to G1 transition), such as MAP-2 kinase, casein kinase II and S6 kinase II, have counterparts that are activated during M-phase in maturing sea star oocytes.


Growth mechanism of titanium dioxide nanowires for dye-sensitized solar cells.
Mesoporous films made of titanium dioxide nanowires are desirable for dye-sensitized solar cells because nanowires provide direct conduction pathways for photogenerated electrons. Anatase titanium dioxide nanowires with polycrystalline microstructure were synthesized on titanium foil using a three-step process. First, the top surface of the titanium foil was transformed to Na(2)Ti(2)O(4)(OH)(2) nanotubes through hydrothermal oxidation in NaOH. Next, the Na(2)Ti(2)O(4)(OH)(2) nanotubes were converted to H(2)Ti(2)O(4)(OH)(2) nanotubes by ion exchange. Finally, the H(2)Ti(2)O(4)(OH)(2) nanotubes were converted to polycrystalline anatase nanowires through a topotactic transformation. The film morphology evolution, crystal structure transformations and growth mechanism are described in detail. Titanium foil reacts with NaOH to form Na(2)Ti(2)O(4)(OH)(2) sheets, which exfoliate and spiral into nanotubes. The Na(2)Ti(2)O(4)(OH)(2) nanotubes are immersed in HCl solution to replace the Na(+) ions with H(+) ions. During the topotactic transformation of H(2)Ti(2)O(4)(OH)(2) nanotubes to anatase TiO(2) nanowires, the sheets made of edge bonded TiO(6) octahedra in the H(2)Ti(2)O(4)(OH)(2) nanotubes dehydrate and move towards each other to form anatase crystals oriented along the nanotube axis which creates a polycrystalline nanowire. These mesoporous TiO(2) nanowire films were suitable for use as dye-sensitized solar cell photoanodes.


Canadian Cardiovascular Society/Canadian Anesthesiologists' Society/Canadian Heart Rhythm Society joint position statement on the perioperative management of patients with implanted pacemakers, defibrillators, and neurostimulating devices.
There are more than 200,000 Canadians living with permanent pacemakers or implantable defibrillators, many of whom will require surgery or invasive procedures each year. They face potential hazards when undergoing surgery; however, with appropriate planning and education of operating room personnel, adverse device-related outcomes should be rare. This joint position statement from the Canadian Cardiovascular Society (CCS) and the Canadian Anesthesiologists' Society (CAS) has been developed as an accessible reference for physicians and surgeons, providing an overview of the key issues for the preoperative, intraoperative, and postoperative care of these patients. The document summarizes the limited published literature in this field, but for most issues, relies heavily on the experience of the cardiologists and anesthesiologists who contributed to this work. This position statement outlines how to obtain information about an individual's type of pacemaker or implantable defibrillator and its programming. It also stresses the importance of determining if a patient is highly pacemaker-dependent and proposes a simple approach for nonelective evaluation of dependency. Although the document provides a comprehensive list of the intraoperative issues facing these patients, there is a focus on electromagnetic interference resulting from electrocautery and practical guidance is given regarding the characteristics of surgery, electrocautery, pacemakers, and defibrillators which are most likely to lead to interference. The document stresses the importance of preoperative consultation and planning to minimize complications. It reviews the relative merits of intraoperative magnet use vs reprogramming of devices and gives examples of situations where one or the other approach is preferable.


Tunable optical delay via carrier induced exciton dephasing in semiconductor quantum wells.
We report the experimental realization of a tunable optical delay by exploiting unique incoherent nonlinear optical processes in semiconductors. The tunable optical delay takes advantage of the strong Coulomb interactions between excitons and free carriers and uses optical injection of free carriers to broaden and bleach an exciton absorption resonance. Fractional delay exceeding 200% has been obtained for an 8 ps optical pulse propagating near the heavy-hole excitonic transition in a GaAs quantum well structure. Tunable optical delay based on optical injection of free carriers avoids strong absorption of the pump beam and is also robust against variations in the frequency of the pump beam.


The lymphocyte function-associated antigen 1 I domain is a transient binding module for intercellular adhesion molecule (ICAM)-1 and ICAM-3 in hydrodynamic flow.
The I domain of lymphocyte function-associated antigen (LFA)-1 contains an intercellular adhesion molecule (ICAM)-1 and ICAM-3 binding site, but the relationship of this site to regulated adhesion is unknown. To study the adhesive properties of the LFA-1 I domain, we stably expressed a GPI-anchored form of this I domain (I-GPI) on the surface of baby hamster kidney cells. I-GPI cells bound soluble ICAM-1 (sICAM-1) with a low avidity and affinity. Flow cell experiments demonstrated a specific rolling interaction of I-GPI cells on bilayers containing purified full length ICAM-1 or ICAM-3. The LFA-1 activating antibody MEM-83, or its Fab fragment, decreased the rolling velocity of I-GPI cells on ICAM-1-containing membranes. In contrast, the interaction of I-GPI cells with ICAM-3 was blocked by MEM-83. Rolling of I-GPI cells was dependent on the presence of Mg2+. Mn2+ only partially substituted for Mg2+, giving rise to a small fraction of rolling cells and increased rolling velocity. This suggests that the I domain acts as a transient, Mg2+-dependent binding module that cooperates with another Mn2+-stimulated site in LFA-1 to give rise to the stable interaction of intact LFA-1 with ICAM-1.


Bone healing induced by local delivery of an engineered parathyroid hormone prodrug.
Regenerative medicine requires innovative therapeutic designs to accommodate high morphogen concentrations in local depots, provide their sustained presence, and enhance cellular invasion and directed differentiation. Here we present an example for inducing local bone regeneration with a matrix-bound engineered active fragment of human parathyroid hormone (PTH(1-34)), linked to a transglutaminase substrate for binding to fibrin as a delivery and cell-invasion matrix with an intervening plasmin-sensitive link (TGplPTH(1-34)). The precursor form displays very little activity and signaling to osteoblasts, whereas the plasmin cleavage product, as it would be induced under the enzymatic influence of cells remodeling the matrix, was highly active. In vivo animal bone-defect experiments showed dose-dependent bone formation using the PTH-fibrin matrix, with evidence of both osteoconductive and osteoinductive bone-healing mechanisms. Results showed that this PTH-derivatized matrix may have potential utility in humans as a replacement for bone grafts or to repair bone defects.


Prolactin signalling to milk protein secretion but not to gene expression depends on the integrity of the Golgi region.
Prolactin added to the incubation medium of lactating mammary epithelial cells is transported from the basal to the apical region of cells through the Golgi region and concomitantly stimulates arachidonic acid release and protein milk secretion. We report that when PRL is added after disorganisation of the Golgi apparatus by brefeldin A treatment, prolactin signalling to expression of genes for milk proteins and prolactin endocytosis are not affected. However, prolactin transport to the apical region of cells (transcytosis), as well as prolactin-induced arachidonic acid release and subsequent stimulation of the secretion of caseins, which are located in a post-Golgi compartment, are inhibited. This inhibition was not a consequence of damage to the secretory machinery, as under the same conditions, protein secretion could be stimulated by the addition of arachidonic acid to the incubation medium. Thus, it is possible to discriminate between prolactin-induced actions that are dependent (signalling to milk protein secretion) or independent (signalling to milk gene expression) on the integrity of the Golgi apparatus. These results suggest that these two biological actions may be transduced via distinct intracellular pathways, and support the hypothesis that prolactin signals may be emitted at various cellular sites.


Coagulation and fibrinolytic activity of cerebrospinal fluid.
Fibrin/fibrinogen degradation products (fragments D and E) were detected in cerebrospinal fluid in 23.4% of 252 patients admitted to a neurological/neurosurgical unit. Other coagulation proteins of low molecular weight (plasminogen and factor IX) were also present but larger proteins (fibrinogen and factor V) were not. These findings are consistent with protein leakage across a blood-CSF barrier damaged by inflammatory, vascular, or neoplastic disease. Fibrin/fibrinogen degradation products in cerebrospinal fluid after subarachnoid haemorrhage may not, therefore, be a reliable index of increased fibrinolytic activity in the subarachnoid space and may be misleading when selecting patients for fibrinolytic blockade.


Overexpression of yeast homologs of the mammalian checkpoint gene RCC1 suppresses the class of alpha-tubulin mutations that arrest with excess microtubules.
Microtubules in eukaryotic cells participate in a variety of nuclear and cytoplasmic structures, reflecting functional requirements and cell cycle position. We are studying the cellular regulation of microtubule assembly and organization in the yeast Saccharomyces cerevisiae. We screened for genes that when overexpressed suppress the growth phenotype of conditional mutants in alpha-tubulin that arrest with excess microtubules at the nonpermissive temperature (class 2 mutations). Here we describe one such suppressing element, called ATS1 (for Alpha Tubulin Suppressor). Overexpression of this gene rescues both the growth and microtubule phenotypes of all class 2 mutations, but not the cold-sensitive mutations that arrest with no microtubules (class 1 mutations). Deletion of ATS1 confers a modest slow growth phenotype which is slightly enhanced in strains containing both a deletion of ATS1 and a class 2 tub 1 mutation. The predicted ATS1 protein contains 333 amino acids and has considerable structural homology to the products of both the mammalian mitotic control gene RCC1 and the S. cerevisiae gene SRM1/PRP20. Overexpression of SRM1/PRP20 also suppresses class 2 mutants. The results suggest that this family of genes may participate in regulatory interactions between microtubules and the cell cycle.


Benzene toxicokinetics in humans: exposure of bone marrow to metabolites.
A three compartment physiologically based toxicokinetic model was fitted to human data on benzene disposition. Two separate groups of model parameter derivations were obtained, depending on which data sets were being fitted. The model was then used to simulate five environmental or occupational exposures. Predicted values of the total bone marrow exposure to benzene and cumulative quantity of metabolites produced by the bone marrow were generated for each scenario. The relation between cumulative quantity of metabolites produced by the bone marrow and continuous benzene exposure was also investigated in detail for simulated inhalation exposure concentrations ranging from 0.0039 ppm to 150 ppm. At the level of environmental exposures, no dose rate effect was found for either model. The occupational exposures led to only slight dose rate effects. A 32 ppm exposure for 15 minutes predicted consistently higher values than a 1 ppm exposure for eight hours for the total exposure of bone marrow to benzene and the cumulative quantity of metabolites produced by the bone marrow. The general relation between the cumulative quantity of metabolites produced by the bone marrow and the inhalation concentration of benzene is not linear. An inflection point exists in some cases leading to a slightly S shaped curve. At environmental levels (0.0039-10 ppm) the curve bends upward, and it saturates at high experimental exposures (greater than 100 ppm).


The prevalence of excessive daytime sleepiness among academic physicians and its impact on the quality of life and occupational performance.
Sleep disorders can affect health and occupational performance of physicians as well as outcomes in patients. The purpose of this study was to assess the prevalence of excessive daytime sleepiness (EDS) measured by the Epworth Sleepiness Scale (ESS) among academic physicians at a tertiary academic medical center in an urban area in the northwest region of Turkey, and to establish a relationship between the self-perceived sleepiness and the quality of life using the EuroQol-5 dimensions (EQ-5D). A questionnaire prepared by the researchers after scanning the literature on the subject was e-mailed to the academic physicians of a tertiary academic medical center in Istanbul. The ESS and the EQ-5D were also included in the survey. The e-mail database of the institution directory was used to compile a list of active academic physicians who practiced clinical medicine. Paired and independent t tests were used for the data analysis at a significance level of p < 0.05. Three hundred and ninety six academic physicians were e-mailed and a total of 252 subjects replied resulting in a 63.6% response rate. There were 84 (33.3%) female and 168 (66.7%) male academic physicians participating in the study. One hundred and eight out of 252 (42.8%) academic physicians were taking night calls (p < 0.001). Ninety study subjects (35.7%) felt they had enough sleep and 84 (33.3%) reported napping daily (p < 0.001). In our sample, 28.6% (N = 72) of the physicians felt sleepy during the day (ESS score > 10) (p < 0.001). In the case of the EQ-5D index and visual analogue scale of the EQ-5D questionnaire (EQ-5D VAS), the status of sleepiness of academic physicians was associated with a poorer quality of life (p < 0.001). More than a 1/4 of the academic physicians suffered from sleepiness. There was an association between the poor quality of life and daytime sleepiness. There was also a positive relationship between habitual napping and being sleepy during the day.


Patterns of change in high frequency precipitation variability over North America.
Precipitation variability encompasses attributes associated with the sequencing and duration of events of the full range of magnitudes. However, climate change studies have largely focused on extreme events. Using analyses of long-term weather station data, we show that high frequency events, such as fraction of wet days in a year and average duration of wet and dry periods, are undergoing significant changes across North America. Further, these changes are more prevalent and larger than those associated with extremes. Such trends also exist for events of a range of magnitudes. Existence of localized clusters with opposing trend to that of broader geographic variation illustrates the role of microclimate and other drivers of trends. Such hitherto unknown patterns over the entire North American continent have the potential to significantly inform our characterization of the resilience and vulnerability of a broad range of ecosystems and agricultural and socio-economic systems. They can also set new benchmarks for climate model assessments.


Implicit learning in individuals with autism spectrum disorders: a meta-analysis.
Individuals with autism spectrum disorders (ASDs) are characterized by social communication difficulties and behavioural rigidity. Difficulties in learning from others are one of the most devastating features of this group of conditions. Nevertheless, the nature of learning difficulties in ASDs is still unclear. Given the relevance of implicit learning for social and communicative functioning, a link has been hypothesized between ASDs and implicit learning deficit. However, studies that have employed formal testing of implicit learning in ASDs provided mixed results. We undertook a systematic search of studies that examined implicit learning in ASDs using serial reaction time (SRT), alternating serial reaction time (ASRT), pursuit rotor (PR), and contextual cueing (CC) tasks, and synthesized the data using meta-analysis. A total of 11 studies were identified, representing data from 407 individuals with ASDs and typically developing comparison participants. The results indicate that individuals with ASDs do not differ in any task considered [SRT and ASRT task: standardized mean difference (SMD) -0.18, 95% confidence interval (CI) -0.71 to 0.36; PR task: SMD -0.34, 95% CI -1.04 to 0.36; CC task: SMD 0.27, 95% CI -0.07 to 0.60]. Based on our synthesis of the existing literature, we conclude that individuals with ASDs can learn implicitly, supporting the hypothesis that implicit learning deficits do not represent a core feature in ASDs.


Potency assays for novel T-cell-inducing vaccines against malaria.
The complex life cycle of the malaria parasite involves several developmental stages in the mammalian host that provide opportunities for vaccine intervention. The effector arm of the immune response that protects against malaria is specific for each stage of the parasite life cycle. While CD4+ and CD8+ T-cells are required to mediate protection during the liver stage, antibodies play a major role before parasite entry into the liver and during the blood stage. Induction of cytotoxic T-cells or those producing IFNgamma has become a major goal for T-cell-inducing vaccines, and the liver stage is currently one of the preferred targets for malaria vaccine development. T-cells can effectively be primed by several vaccine strategies, including recombinant vectors. A wide range of such vectors is currently available and their use alone or in combination induces high frequencies of antigen-specific T-cells in animal models and humans. Therefore, traditional potency assays, such as the highly sensitive ex vivo and cultured ELISPOT, are being complemented by less sensitive, but more flexible techniques, such as flow cytometry, which allows characterization of multifunctional antigen-specific T-cells following vaccination.


A comparative study of ERG status assessment on DNA, mRNA, and protein levels using unique samples from a Swedish biopsy cohort.
The ERG rearrangement is identified in approximately 50% of prostate cancer screened cohorts and is known to be highly specific. This genetic aberration, most commonly leading to the TMPRSS2-ERG fusion, but also SLC45A3-ERG or NDRG1-ERG fusions, all leading to an overexpression of a truncated ERG protein. Most studies have applied in situ hybridization (FISH) methods or mRNA-based assays to investigate the ERG status. Recently, studies showed that ERG protein levels assessed by ERG antibodies can be used as a surrogate marker for ERG rearrangement. In the current study, we investigate ERG status on a series of diagnostic biopsies using DNA-based, mRNA-based, and protein-based assays. We formally compared 3 assay results (ie, FISH, fusion mRNA, and immunohistochemistry) to identify which method could be most appropriate to use when having limited amount of tissue. ERG rearrangement was found in 56% of the cases. Comparing ERG rearrangement status by FISH with ERG overexpression and TMPRSS2-ERG fusion transcript we found 95.1% (154/162, Fisher exact test 9.50E-36) and 85.2% (138/162, Fisher exact test 7.26E-22) concordance, respectively. We show that the ERG antibody highly correlates with the ERG rearrangement with high sensitivity and specificity. We also identified the most common TMPRSS2-ERG isoform in the majority of ERG rearranged cases. These results provide compelling evidence that the ERG antibody can be used to further investigate the role of ERG in prostate cancer.


Data-based reconstruction of complex geospatial networks, nodal positioning and detection of hidden nodes.
Given a complex geospatial network with nodes distributed in a two-dimensional region of physical space, can the locations of the nodes be determined and their connection patterns be uncovered based solely on data? We consider the realistic situation where time series/signals can be collected from a single location. A key challenge is that the signals collected are necessarily time delayed, due to the varying physical distances from the nodes to the data collection centre. To meet this challenge, we develop a compressive-sensing-based approach enabling reconstruction of the full topology of the underlying geospatial network and more importantly, accurate estimate of the time delays. A standard triangularization algorithm can then be employed to find the physical locations of the nodes in the network. We further demonstrate successful detection of a hidden node (or a hidden source or threat), from which no signal can be obtained, through accurate detection of all its neighbouring nodes. As a geospatial network has the feature that a node tends to connect with geophysically nearby nodes, the localized region that contains the hidden node can be identified.


When the Moon had a magnetosphere.
Apollo lunar samples reveal that the Moon generated its own global magnetosphere, lasting from ~4.25 to ~2.5 billion years (Ga) ago. At peak lunar magnetic intensity (4 Ga ago), the Moon was volcanically active, likely generating a very tenuous atmosphere, and, it is believed, was at a geocentric distance of ~18 Earth radii (R E). Solar storms strip a planet's atmosphere over time, and only a strong magnetosphere would be able to provide maximum protection. We present simplified magnetic dipole field modeling confined within a paraboloidal-shaped magnetopause to show how the expected Earth-Moon coupled magnetospheres provide a substantial buffer from the expected intense solar wind, reducing Earth's atmospheric loss to space.


Cloning of self-major histocompatibility complex antigen-specific suppressor cells from adult bone marrow.
We examined if suppressor cell clones may be established from adult bone marrow that contains a population of cells capable of specifically downregulating the immune response directed toward self-major histocompatibility complex (MHC) antigens. Freshly prepared adult C3H (H-2k) marrow cells were cultured in medium containing interleukin 2 (IL-2), IL-3, or a mixture of IL-2 and IL-3. After 7-10 d, cells grown in IL-3-containing medium were screened for their capacity to suppress cytotoxic T lymphocyte (CTL) generation against self-MHC antigens in allogeneic mixed lymphocyte cultures. Cells capable of suppressing anti-C3H CTL generation were cloned by limiting dilution. Several suppressor clones were established that exhibited strong suppression of anti-H-2k, anti-H-2Kk/Ik, and anti-H-2Dk CTL generation, but failed to suppress anti-H-2d and anti-H-2b responses. When tested in a skin allograft model, intravenous injections of these bone marrow-derived anti-self suppressor cells (2.5 x 10(7) cells) together with IL-3 induced prolongation of C3H skin allografts in anti-mouse lymphocyte serum-treated B6AF1 mice. Injection of IL-3 alone had no effect on allograft survival. Moreover, these cells failed to prolong B10.AKM skin allografts on B6AF1 recipients. Northern blot analysis showed that these cells express full-length transcripts of the T cell receptor (TCR) gamma gene, but not those of TCR alpha, beta, or delta genes. However, no rearrangement of gamma gene was observed by Southern blot analysis. Flow cytometric analysis revealed that bone marrow-derived suppressor cells are strongly positive for Thy-1 antigen but negative for CD3, CD4 (L3T4), and CD8 (Lyt-2) surface markers, and express only class I MHC antigens. Suppressor cells derived from adult bone marrow may play an important role in extrathymic induction of self-tolerance.


The clinical features of the piriformis syndrome: a systematic review.
Piriformis syndrome, sciatica caused by compression of the sciatic nerve by the piriformis muscle, has been described for over 70 years; yet, it remains controversial. The literature consists mainly of case series and narrative reviews. The objectives of the study were: first, to make the best use of existing evidence to estimate the frequencies of clinical features in patients reported to have PS; second, to identify future research questions. A systematic review was conducted of any study type that reported extractable data relevant to diagnosis. The search included all studies up to 1 March 2008 in four databases: AMED, CINAHL, Embase and Medline. Screening, data extraction and analysis were all performed independently by two reviewers. A total of 55 studies were included: 51 individual and 3 aggregated data studies, and 1 combined study. The most common features found were: buttock pain, external tenderness over the greater sciatic notch, aggravation of the pain through sitting and augmentation of the pain with manoeuvres that increase piriformis muscle tension. Future research could start with comparing the frequencies of these features in sciatica patients with and without disc herniation or spinal stenosis.


Demands of Simulated Commuting Using an Electrically Assisted Bicycle.
The American College of Sports Medicine (ACSM) recommends adults participate in weekly aerobic activity for a minimum of 30 minutes moderate intensity exercise 5 days per week or 20 minutes of vigorous activity 3 days per week. The electrically assisted bicycle may help individuals achieve the ACSM's aerobic recommendations and introduce inactive individuals to physical activity. To compare the physiological requirements of riding a bicycle with electric pedal assist versus non-assist among healthy active young adults. 6 males and 6 females completed two randomized cycling trials using electric pedal assist (PAB) and non-assist (NON). Cycling trials were completed over a 3.54 km course with varying terrain. Time to completion was faster in the PAB (12.5 ± 0.3 min) than the NON (13.8 ± 0.3 min, p=0.01). Rating of Perceived Exertion (RPE) was lower in the PAB (12.0 ± 0.4) than the NON (14.8 ± 0.5, p < 0.001). There was no difference in mean VO2 between PAB (2.3 ± 0.1 L·min-1) and NON (2.5 ± 0.1 L·min-1, p=0.45). There was no difference in mean power output when comparing PAB (115 ± 11 Watts) to NON (128 ± 11 Watts, p=0.38). There was no difference in heart rate between PAB (147 ± 5 bpm) and NON (149 ± 5 bpm, p=0.77). Recreationally active younger (college age) individuals may self-select a similar physiological intensity of physical activity regardless of mechanical assistance, resulting in quicker completion of a commuting task with PAB. Both the PAB and NON exercise bouts met ACSM criteria for vigorous exercise.


The dawn of cognitive genetics? Crucial developmental caveats.
Attempts to bridge genetics and cognition are rapidly coming to the forefront of cognitive neuroscience. It is therefore crucial to evaluate the current state of knowledge about disorders of known genetic origin as a way of assessing whether, and if so how, links between genotype and cognitive phenotype can be drawn, however indirect these links might be. We review recent empirical findings from research on genetic disorders at three levels of description--cognitive, neural systems, and cellular--that caution against simple genotype-phenotype mappings at all levels. Most importantly, interdisciplinary efforts to integrate human genetics and cognition will need to operationalize the mechanisms driving both typical and atypical developmental processes over time.


Photoenhanced degradation of veratraldehyde upon the heterogeneous ozone reactions.
Light-induced heterogeneous reactions between gas-phase ozone and veratraldehyde adsorbed on silica particles were performed. At an ozone mixing ratio of 250 ppb, the loss of veratraldehyde largely increased from 1.81 x 10(-6) s(-1) in the dark to 2.54 x 10(-5) s(-1) upon exposure to simulated sunlight (lambda > 300 nm). The observed rates of degradation exhibited linear dependence with the ozone in the dark ozonolysis experiments which change in the non-linear Langmuir-Hinshelwood dependence in the experiments with simultaneous ozone and light exposure of the coated particles. When the coated silica particles were exposed only to simulated sunlight in absence of ozone the loss of veratraldehyde was about three times higher i.e. 5.97 x 10(-6) s(-1) in comparison to the ozonolysis experiment under dark conditions at 250 ppb ozone mixing ratio, 1.81 x 10(-6) s(-1).These results clearly show that the most important loss of veratraldehyde occurs under simultaneous ozone and light exposure of the coated silica particles. The main identified product in the heterogeneous reactions between gaseous ozone and adsorbed veratraldehyde under dark conditions and in presence of light was veratric acid.Carbon yields of veratric acid were calculated and the obtained results indicated that at low ozone mixing ratio (250 ppb) the carbon yield obtained under dark conditions is 70% whereas the carbon yield obtained in the experiments with simultaneous ozone and light exposure of the coated particles is 40%. In both cases the carbon yield of veratric acid exponentially decayed leading to the plateau ( approximately 35% of carbon yield) at an ozone mixing ratio of 6 ppm. Two reaction products i.e. 3-hydroxy-4-methoxybenzoic acid and 4-hydroxy-3-methoxybenzoic acid were identified (confirmed with the standards) only in the experiments performed under simultaneous ozonolysis and light irradiation of the particles.


Surface-emitting circular DFB, disk-, and ring-Bragg resonator lasers with chirped gratings. II: nonuniform pumping and far-field patterns.
This is a continuation of our previous work [Opt. Express 16, 9155 (2008)]. In this paper we investigate the effect of nonuniform pumping on the modal properties of surface-emitting chirped circular grating lasers. By numerically solving the coupled-mode equations and matching the boundaries we compare and discuss the threshold pump levels and frequency detuning factors for three pumping profiles: uniform, Gaussian, and annular. Depending on the overlap of the pumping and modal profiles, Gaussian pumping results in the lowest threshold pump levels except for the fundamental mode of ring Bragg resonator laser, and annular pumping provides larger threshold discrimination between the fundamental and first-order modes of circular DFB and ring Bragg resonator lasers, which is favorable for single-mode operation in these lasers. We also study the far-field patterns of the fundamental modes of circular DFB, disk-, and ring- Bragg resonator lasers. Circular DFB and ring Bragg resonator lasers have the first-order dominating peak, while disk Bragg resonator laser exhibits the zeroth-order dominating peak.


Single nucleotide seed modification restores in vivo tolerability of a toxic artificial miRNA sequence in the mouse brain.
Huntington's disease is a fatal neurodegenerative disease caused by polyglutamine-expansion in huntingtin (HTT). Recent work showed that gene silencing approaches, including RNA interference (RNAi), improve disease readouts in mice. To advance RNAi to the clinic, we designed miHDS1, with robust knockdown of human HTT and minimized silencing of unintended transcripts. In Rhesus macaque, AAV delivery of miHDS1 to the putamen reduced HTT expression with no adverse effects on neurological status including fine and gross motor skills, no immune activation and no induction of neuropathology out to 6 weeks post injection. Others showed safety of a different HTT-targeting RNAi in monkeys for 6 months. Application of miHDS1 to Huntington's patients requires further safety testing in normal rodents, despite the fact that it was optimized for humans. To satisfy this regulatory requirement, we evaluated normal mice after AAV.miHDS1 injection. In contrast to monkeys, neurological deficits occurred acutely in mice brain and was attributed to off-target silencing through interactions of miHDS1 with the 3'UTR of other transcripts. While we resolved miHDS1 toxicity in mouse brain and maintained miHDS1-silencing efficacy, these studies highlight that optimizing nucleic acid-based medicines for safety in humans presents challenges for safety testing in rodents or other distantly related species.


Cell contact-dependent outer membrane exchange in myxobacteria: genetic determinants and mechanism.
Biofilms are dense microbial communities. Although widely distributed and medically important, how biofilm cells interact with one another is poorly understood. Recently, we described a novel process whereby myxobacterial biofilm cells exchange their outer membrane (OM) lipoproteins. For the first time we report here the identification of two host proteins, TraAB, required for transfer. These proteins are predicted to localize in the cell envelope; and TraA encodes a distant PA14 lectin-like domain, a cysteine-rich tandem repeat region, and a putative C-terminal protein sorting tag named MYXO-CTERM, while TraB encodes an OmpA-like domain. Importantly, TraAB are required in donors and recipients, suggesting bidirectional transfer. By use of a lipophilic fluorescent dye, we also discovered that OM lipids are exchanged. Similar to lipoproteins, dye transfer requires TraAB function, gliding motility and a structured biofilm. Importantly, OM exchange was found to regulate swarming and development behaviors, suggesting a new role in cell-cell communication. A working model proposes TraA is a cell surface receptor that mediates cell-cell adhesion for OM fusion, in which lipoproteins/lipids are transferred by lateral diffusion. We further hypothesize that cell contact-dependent exchange helps myxobacteria to coordinate their social behaviors.


Whole-body hyperthermia combined with hyperthermic intraperitoneal chemotherapy for the treatment of stage IV advanced gastric cancer.
Currently known as a metastatic disease, stage IV gastric cancer cannot be cured by surgery, but treatments are recommended to relieve symptoms such as pain and to prolong survival. With better access to metastases and certain large or inoperable tumours, we applied two treatment sessions of combined therapy of whole-body hyperthermia and hyperthermic intraperitoneal chemo-perfusion in the treatment group, while patients in the control group were treated with oxaliplatin combined with 5-fluorouracil chemotherapy or Xeloda. We used the RECIST criteria for outcome evaluation. With the combined treatment, we found the complete and partial remission rate of patients to be 61.5%, and the rate of stable disease was 19.2%. Symptoms such as pain and a large volume of ascites were alleviated, and the quality of life was correspondingly improved. In addition, the combined treatment had a significant therapeutic benefit against the primary tumour and the metastases to the lymph nodes and liver. Survival time was also significantly prolonged (the 1-year survival rate was 38.5% compared to the control group rate of 19%). These results suggest that whole-body hyperthermia combined with hyperthermic intraperitoneal chemotherapy is an effective treatment for patients with advanced gastric malignancies.


Application of fluorescence resonance energy transfer and magnetic twisting cytometry to quantify mechanochemical signaling activities in a living cell.
Mechanotransduction is the process by which living cells sense mechanical forces and then convert them into biochemical signaling. Recently, we showed that mechanical stress is transduced from the cell surface to remote cytoplasmic sites within 0.3 seconds, which is at least 40 to 50 times faster than soluble factor-induced signal transduction, and the sites of mechanotransduction colocalize with mechanical stress-induced microtubule displacements. These results suggest that mechanotransduction employs mechanisms different from those of soluble factor-induced signal transduction. Here, we describe a protocol that utilizes fluorescence resonance energy transfer (FRET) and a magnetic twisting cytometry (MTC) device to capture rapid mechanochemical signaling activities in living cells.


First trimester bleeding.
Vaginal bleeding in the first trimester occurs in about one fourth of pregnancies. About one half of those who bleed will miscarry. Guarded reassurance and watchful waiting are appropriate if fetal heart sounds are detected, if the patient is medically stable, and if there is no adnexal mass or clinical sign of intraperitoneal bleeding. Discriminatory criteria using transvaginal ultrasonography and beta subunit of human chorionic gonadotropin testing aid in distinguishing among the many conditions of first trimester bleeding. Possible causes of bleeding include subchorionic hemorrhage, embryonic demise, anembryonic pregnancy, incomplete abortion, ectopic pregnancy, and gestational trophoblastic disease. When beta subunit of human chorionic gonadotropin reaches levels of 1,500 to 2,000 mIU per mL (1,500 to 2,000 IU per L), a normal pregnancy should exhibit a gestational sac by transvaginal ultrasonography. When the gestational sac is greater than 10 mm in diameter, a yolk sac must be present. A live embryo must exhibit cardiac activity when the crown-rump length is greater than 5 mm. In a normal pregnancy, beta subunit of human chorionic gonadotropin levels increase by 80 percent every 48 hours. The absence of any normal discriminatory findings is consistent with early pregnancy failure, but does not distinguish between ectopic pregnancy and failed intrauterine pregnancy. The presence of an adnexal mass or free pelvic fluid represents ectopic pregnancy until proven otherwise. Medical management with misoprostol is highly effective for early intrauterine pregnancy failure with the exception of gestational trophoblastic disease, which must be surgically evacuated. Expectant treatment is effective for many patients with incomplete abortion. Medical management with methotrexate is highly effective for properly selected patients with ectopic pregnancy. Follow-up after early pregnancy loss should include attention to future pregnancy planning, contraception, and psychological aspects of care.


Hybrid dysgenesis-induced revertants of insertions at the 5' end of the rudimentary gene in Drosophila melanogaster: transposon-induced control mutations.
Mutations in the 5' control region of the rudimentary (r) gene of Drosophila melanogaster were generated by using hybrid dysgenesis to mobilize P elements that were already inserted within this region. Eighteen new mutations out of 7793 chromosomes were isolated. Among the mutations were small insertions, deletions and inversions. All five of the deletions deleted into the 5' coding region of the r gene resulting in a severe mutant phenotype. The inversions and small insertions left the coding region intact, but altered the 5' control region. Northern analyses of the message levels in adult females indicated that these mutations alter the amount of the wild-type rudimentary message. These data also indicated that there is a region about 800 bp upstream of the start of transcription which is necessary for normal r expression. This region is not involved with the temporal and spatial regulation of r gene expression, but with the quantitative expression. An analysis of the DNA changes associated with each of the mutations demonstrates that P elements usually mobilize in an imprecise manner. Of the 22 mutations that have been isolated to date, only three are precise excisions. This means that P elements are potent mutators of the genome.


Proton pump inhibitors and the risk of adverse cardiac events.
Recent evidence suggests that proton pump inhibitors (PPIs) might be linked with adverse cardiac events, but a causal relationship is unproven. We applied the self-matched case series method to two studies using population-based health care data from Ontario, Canada between 1996 and 2008. The first included subjects aged 66 years or older hospitalized for acute myocardial infarction within 12 weeks following initiation of PPI, while the second included subjects hospitalized for heart failure. In both studies we designated the primary risk interval as the initial 4 weeks of therapy and the control interval as the final 4 weeks. To test the specificity of our findings we examined use of histamine H2 receptor antagonists and benzodiazepines, drugs with no plausible causal link to adverse cardiac events. During the 13-year study period, we identified 5550 hospital admissions for acute myocardial infarction and 6003 admissions for heart failure within 12 weeks of commencing PPI therapy. In the main analyses, we found that initiation of a PPI was associated with a higher risk of acute myocardial infarction (odds ratio 1.8; 95% confidence interval 1.7 to 1.9) and heart failure (odds ratio 1.8; 95% confidence interval 1.7 to 1.9). However, secondary analyses revealed similar risk estimates histamine H2 receptor antagonists and benzodiazepines, drugs with no known or suspected association with adverse cardiac events. PPIs are associated with a short-term risk of adverse cardiac events, but similar associations are seen with other drugs exhibiting no known cardiac toxicity. Collectively these observations suggest that the association between PPIs and adverse cardiac events does not represent reflect cause-and-effect.


Inbreeding Affects Gene Expression Differently in Two Self-Incompatible Arabidopsis lyrata Populations with Similar Levels of Inbreeding Depression.
Knowledge of which genes and pathways are affected by inbreeding may help understanding the genetic basis of inbreeding depression, the potential for purging (selection against deleterious recessive alleles), and the transition from outcrossing to selfing. Arabidopsis lyrata is a predominantly self-incompatible perennial plant, closely related to the selfing model species A. thaliana. To examine how inbreeding affects gene expression, we compared the transcriptome of experimentally selfed and outcrossed A. lyrata originating from two Scandinavian populations that express similar inbreeding depression for fitness (∂ ≈ 0.80). The number of genes significantly differentially expressed between selfed and outcrossed individuals were 2.5 times higher in the Norwegian population (≈ 500 genes) than in the Swedish population (≈ 200 genes). In both populations, a majority of genes were upregulated on selfing (≈ 80%). Functional annotation analysis of the differentially expressed genes showed that selfed offspring were characterized by 1) upregulation of stress-related genes in both populations and 2) upregulation of photosynthesis-related genes in Sweden but downregulation in Norway. Moreover, we found that reproduction- and pollination-related genes were affected by inbreeding only in Norway. We conclude that inbreeding causes both general and population-specific effects. The observed common effects suggest that inbreeding generally upregulates rather than downregulates gene expression and affects genes associated with stress response and general metabolic activity. Population differences in the number of affected genes and in effects on the expression of photosynthesis-related genes show that the genetic basis of inbreeding depression can differ between populations with very similar levels of inbreeding depression.


Aerobic scope falls to nil at Pcrit and anaerobic ATP production increases below Pcrit in the tidepool sculpin, Oligocottus maculosus.
The critical oxygen tension of whole-animal oxygen uptake rate, or Pcrit, has historically been defined as the oxygen partial pressure (PO2) at which aerobic scope falls to zero and further declines in PO2 require substrate-level phosphorylation to meet shortfalls in aerobic ATP production, thereby time-limiting survival. Despite the inclusion of aerobic scope and anaerobic ATP production in the definition, little effort has been made to verify that Pcrit measurements, the vast majority of which are obtained using respirometry in resting animals, actually reflect the predictions of zero aerobic scope and a transition to increasing reliance on anaerobic ATP production. To test these predictions, we compared aerobic scope and levels of whole-body lactate at oxygen partial pressures (PO2s) bracketing Pcrit obtained in resting fish during progressive hypoxia in the tidepool sculpin, Oligocottus maculosus. We found that aerobic scope falls to zero at Pcrit and, in resting fish exposed to PO2s < Pcrit, whole-body lactate accumulated pointing to an increased reliance on anaerobic ATP production. These results support the interpretation of Pcrit as a key oxygen threshold at which aerobic scope falls to nil and, below Pcrit, survival is time-limited based on anaerobic metabolic capacity.


Angiotensin II-induced hypertension in the rat. Effects on the plasma concentration, renal excretion, and tissue release of prostaglandins.
We examined in rats the effects of intraperitoneal angiotensin II (AII) infusion for 12 d on urinary excretion, plasma concentration, and in vitro release of prostaglandin (PG) E2 and 6-keto-PGF1 alpha, a PGI2 metabolite. AII at 200 ng/min increased systolic blood pressure (SBP) progressively from 125 +/- 3 to 170 +/- 9 mmHg (P less than 0.01) and elevated fluid intake and urine volume. Urinary 6-keto-PGF1 alpha excretion increased from 38 +/- 6 to 55 +/- 5 and 51 +/- 7 ng/d (P less than 0.05) on days 8 and 11, respectively, of AII infusion, but urinary PGE2 excretion did not change. Relative to a control value of 129 +/- 12 pg/ml in vehicle-infused (V) rats, arterial plasma 6-keto-PGF1 alpha concentration increased by 133% (P less than 0.01) with AII infusion. Aortic rings from AII-infused rats released more 6-keto-PGF1 alpha (68 +/- 7 ng/mg) during 15-min incubation in Krebs solution than did rings from V rats (40 +/- 3 ng/mg); release of PGE2, which was less than 1% of that of 6-keto-PGF1 alpha, was also increased. Slices of inner renal medulla from AII-infused rats released more 6-keto-PGF1 alpha (14 +/- 1 ng/mg) during incubation than did slices from V rats (8 +/- 1 ng/mg, P less than 0.05), but PGE2 release was not altered. In contrast, AII infusion did not alter release of 6-keto-PGF1 alpha or PGE2 from inferior vena cava segments or from renal cortex slices. Infusion of AII at 125 ng/min also increased SBP, plasma 6-keto-PGF1 alpha concentration, and in vitro release of 6-keto-PGF1 alpha from rings of aorta and renal inner medulla slices; at 75 ng/min AII had no effect. SBP on AII infusion day 11 correlated positively with both 6-keto-PGF1 alpha plasma concentration (r = 0.54) and net aortic ring release (r = 0.70) when data from all rats were combined. We conclude that augmentation of PGI2 production is a feature of AII-induced hypertension. The enhancement of PGI2 production may be an expression of nonspecific alteration in vascular structure and metabolic functions during AII-induced hypertension, as well as the result of a specific effect of the peptide on the arachidonate-prostaglandin system.


Functional analysis of the new barley gene HvKu80 indicates that it plays a key role in double-strand DNA break repair and telomere length regulation.
Genotoxic stress causes a reduced stability of the plant genome and has a detrimental effect on plant growth and productivity. Double-strand breaks (DSBs) are the most harmful of all DNA lesions because they cause the loss of genetic information on both strands of the DNA helix. In the presented study the coding and genomic sequences of the HvKu80 gene were determined. A mutational analysis of two fragments of HvKu80 using TILLING (Targeting Induced Local Lesions IN Genomes) allowed 12 mutations to be detected, which resulted in identification of 11 alleles. Multidirectional analyses demonstrated that the HvKu80 gene is involved in the elimination of DSBs in Hordeum vulgare. The barley mutants carrying the identified ku80.c and ku80.j alleles accumulated bleomycin-induced DSBs to a much greater extent than the parent cultivar 'Sebastian'. The altered reaction of the mutants to DSB-inducing agent and the kinetics of DNA repair in these genotypes are associated with a lower expression level of the mutated gene. The study also demonstrated the significant role of the HvKu80 gene in the regulation of telomere length in barley.


Regulation of polymerase exchange between Poleta and Poldelta by monoubiquitination of PCNA and the movement of DNA polymerase holoenzyme.
To ensure efficient and timely replication of genomic DNA, organisms in all three kingdoms of life possess specialized translesion DNA synthesis (TLS) polymerases (Pols) that tolerate various types of DNA lesions. It has been proposed that an exchange between the replicative DNA Pol and the TLS Pol at the site of DNA damage enables lesion bypass to occur. However, to date the molecular mechanism underlying this process is not fully understood. In this study, we demonstrated in a reconstituted system that the exchange of Saccharomyces cerevisiae Poldelta with Poleta requires both the stalling of the holoenzyme and the monoubiquitination of proliferating cell nuclear antigen (PCNA). A moving Poldelta holoenzyme is refractory to the incoming Poleta. Furthermore, we showed that the Poleta C-terminal PCNA-interacting protein motif is required for the exchange process. We also demonstrated that the second exchange step to bring back Poldelta is prohibited when Lys-164 of PCNA is monoubiquitinated. Thus the removal of the ubiquitin moiety from PCNA is likely required for the reverse exchange step after the lesion bypass synthesis by Poleta.


Autoimmunity and tuberculosis. Opposite association with TNF polymorphism.
To examine the influence of the -308 and -238 single nucleotide polymorphisms (SNP) of tumor necrosis factor-a gene (TNF) on patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), primary Sjogren's syndrome (SS), and tuberculosis (TB). Genomic DNA from patients with RA (n = 165), SLE (n = 100), primary SS (n = 67), and TB (n = 135) and ethnically matched controls (n = 430) was genotyped for TNF -308 and -238 SNP by PCR-RFLP. TNF -308A allele was associated with RA (odds ratio, OR 1.8, p = 0.002), SLE (OR 2.6, p < 0.0001), and primary SS (OR 2.9, p < 0.0001). TNF -308G was associated with TB (OR 1.8, p = 0.02). The -308 GG genotype was protective for autoimmunity (p < 0.003). TNF -238A allele was protective for autoimmunity but represented a susceptibility factor for TB (OR 2.2, p < 0.0001). Haplotype -308A-238G was a protective factor against TB, whereas it carried susceptibility for RA, SLE, and primary SS (p < 0.0001). The results show an opposite association of TNF polymorphism with autoimmunity and TB, and suggest the existence of heterozygote advantage, sustaining the hypothesis that autoimmune diseases are a consequence of natural selection for enhanced TB resistance. Data also provide genetic evidence supporting the common variants/multiple disease hypothesis, which emphasizes that many disease genes may not be disease-specific, and that similar immunogenetic mechanisms underlie autoimmune diseases.


Whole genome analysis of water buffalo and global cattle breeds highlights convergent signatures of domestication.
More people globally depend on the water buffalo than any other domesticated species, and as the most closely related domesticated species to cattle they can provide important insights into the shared evolutionary basis of domestication. Here, we sequence the genomes of 79 water buffalo across seven breeds and compare patterns of between breed selective sweeps with those seen for 294 cattle genomes representing 13 global breeds. The genomic regions under selection between cattle breeds significantly overlap regions linked to stature in human genetic studies, with a disproportionate number of these loci also shown to be under selection between water buffalo breeds. Investigation of potential functional variants in the water buffalo genome identifies a rare example of convergent domestication down to the same mutation having independently occurred and been selected for across domesticated species. Cross-species comparisons of recent selective sweeps can consequently help identify and refine important loci linked to domestication.


Phosphorylation of tau is regulated by PKN.
For the phosphorylation state of microtubule-associated protein, tau plays a pivotal role in regulating microtubule networks in neurons. Tau promotes the assembly and stabilization of microtubules. The potential for tau to bind to microtubules is down-regulated after local phosphorylation. When we investigated the effects of PKN activation on tau phosphorylation, we found that PKN triggers disruption of the microtubule array both in vitro and in vivo and predominantly phosphorylates tau in microtubule binding domains (MBDs). PKN has a catalytic domain highly homologous to protein kinase C (PKC), a kinase that phosphorylates Ser-313 (= Ser-324, the number used in this study) in MBDs. Thus, we identified the phosphorylation sites of PKN and PKC subtypes (PKC-alpha, -betaI, -betaII, -gamma, -delta, -epsilon, -zeta, and -lambda) in MBDs. PKN phosphorylates Ser-258, Ser-320, and Ser-352, although all PKC subtypes phosphorylate Ser-258, Ser-293, Ser-324, and Ser-352. There is a PKN-specific phosphorylation site, Ser-320, in MBDs. HIA3, a novel phosphorylation-dependent antibody recognizing phosphorylated tau at Ser-320, showed immunoreactivity in Chinese hamster ovary cells expressing tau and the active form of PKN, but not in Chinese hamster ovary cells expressing tau and the inactive form of PKN. The immunoreactivity for phosphorylated tau at Ser-320 increased in the presence of a phosphatase inhibitor, FK506 treatment, which means that calcineurin (protein phosphatase 2B) may be involved in dephosphorylating tau at Ser-320 site. We also noted that PKN reduces the phosphorylation recognized by the phosphorylation-dependent antibodies AT8, AT180, and AT270 in vivo. Thus PKN serves as a regulator of microtubules by specific phosphorylation of tau, which leads to disruption of tubulin assembly.


Experiencing breast cancer in managed care.
The interest of managed care organizations (MCOs) in decreasing care and outcome variance and lowering costs has created many concerns including those pertaining to the complex and costly nature of cancer care. In this study, we used a hermeneutic phenomenological approach involving semistructured interviews of 14 women with breast cancer to examine MCO enrollees' experiences related to their cancer treatment. Results comprise two themes: managed care tasks and managing or mediating between the MCO and the cancer. These themes, and their accompanying categories, depict the increased burden encountered by women experiencing breast cancer in a managed care system who perceived that they were solely responsible for mediating between their care providers and MCO. Though enrolled in less-restrictive MCOs, participants expressed many difficulties in managing dissimilar and disconnected demands posed by the cancer and the MCO. These findings might serve as a basis for policy and practice changes to decrease this real patient burden.


Fabrication of microfluidic cavities using Si-to-glass anodic bonding.
We demonstrate the fabrication of ∼1.08 μm deep microfluidic cavities with characteristic size as large as 7 mm × 11 mm or 11 mm diameter, using a silicon-glass anodic bonding technique that does not require posts to act as separators to define cavity height. Since the phase diagram of 3He is significantly altered under confinement, posts might act as pinning centers for phase boundaries. The previous generation of cavities relied on full wafer-bonding which is more prone to failure and requires dicing post-bonding, whereas these cavities are made by bonding a pre-cut piece of Hoya SD-2 glass to a patterned piece of silicon in which the cavity is defined by etching. Anodic bonding was carried out at 425 °C with 200 V, and we observe that pressurizing the cavity to failure (>30 bars pressure) results in glass breaking, rather than the glass-silicon bond separation. In this article, we discuss the detailed fabrication of the cavity, its edges, and details of the junction between the coin silver fill line and the silicon base of the cavity that enables a low internal-friction joint. This feature is important for mass coupling torsional oscillator experimental assays of the superfluid inertial contribution where a high quality factor (Q) improves frequency resolution. The surface preparation that yields well-characterized smooth surfaces to eliminate pinning sites, the use of transparent glass as a cover permitting optical access, low temperature capability, and attachment of pressure-capable ports for fluid access may be features that are important in other applications.


Mechanisms of evoked and induced responses in MEG/EEG.
Cortical responses, recorded by electroencephalography and magnetoencephalography, can be characterized in the time domain, to study event-related potentials/fields, or in the time-frequency domain, to study oscillatory activity. In the literature, there is a common conception that evoked, induced, and on-going oscillations reflect different neuronal processes and mechanisms. In this work, we consider the relationship between the mechanisms generating neuronal transients and how they are expressed in terms of evoked and induced power. This relationship is addressed using a neuronally realistic model of interacting neuronal subpopulations. Neuronal transients were generated by changing neuronal input (a dynamic mechanism) or by perturbing the systems coupling parameters (a structural mechanism) to produce induced responses. By applying conventional time-frequency analyses, we show that, in contradistinction to common conceptions, induced and evoked oscillations are perhaps more related than previously reported. Specifically, structural mechanisms normally associated with induced responses can be expressed in evoked power. Conversely, dynamic mechanisms posited for evoked responses can induce responses, if there is variation in neuronal input. We conclude, it may be better to consider evoked responses as the results of mixed dynamic and structural effects. We introduce adjusted power to complement induced power. Adjusted power is unaffected by trial-to-trial variations in input and can be attributed to structural perturbations without ambiguity.


Intralesional photocoagulation of periorbital hemangiomas.
The purpose of this study was to examine the effects of intralesional bare fiber photocoagulation with the KTP and Nd:YAG lasers on periorbital hemangiomas of infancy. Initial reports by Apfelberg and Gregory suggest that intralesional laser therapy may have a role in the treatment of hemangiomas. Intralesional photocoagulation may be preferred to superficial laser treatment for several reasons. It may decrease cutaneous skin damage and more effectively reduce bulky, deep lesions. Twenty-three patients with periorbital hemangiomas were treated (KTP, n = 7; Nd:YAG, n = 16). An 18-g Angiocath was placed into the lesion to pass the fiber through. Laser energy was delivered by means of a 0.6-mm bare fiber at 10 to 15 J (KTP) or 7 J (Nd:YAG). Treatments were done under general anesthesia. Patients were followed closely for 1 month and then monthly to assess results and complications. Results are as follows: 61 percent demonstrated 50-percent or more reduction at 3 months; 22 percent demonstrated 50-percent or more reduction in 3 to 8 months, i.e., 83 percent of patients had 50-percent or more reduction within 8 months. To achieve these results, two patients required two treatments. The remaining 17 percent had 10- to 14-percent reduction at 1 to 3 months. Two of these patients had two treatments. A subgroup of patients had a very dramatic response. Thirty-five percent (8 of 23) had 50- to 90-percent reduction in 1 month. It is unclear why these patients responded so dramatically. We expected some ulceration during the healing phase. Seventeen percent developed ulceration. Complications were limited (4 percent) to one wound infection. Intralesional photocoagulation treatment with the KTP and Nd:YAG lasers is effective and safe for the treatment of periorbital hemangiomas in the majority of patients with minimal complications. Further study is necessary to identify factors that result in dramatic or limited responses.


Biomarkers in Neuropsychiatry: A Prospect for the Twenty-First Century?
The search for biomarkers to aid in the diagnosis and prognosis of psychiatric conditions and predict response to treatment is a focus of twenty-first century medicine. The current lack of biomarkers in routine use is attributable in part to the existing way mental health conditions are diagnosed, being based upon descriptions of symptoms rather than causal biological evidence. New ways of conceptualizing mental health disorders together with the enormous advances in genetic, epidemiological, and neuroscience research are informing the brain circuits and physiological mechanisms underpinning behavioural constructs that cut across current diagnostic DSM-5 categories. Combining these advances with 'Big Data', analytical approaches offer new opportunities for biomarker development. Here we provide an introductory perspective to this volume, highlighting methodological strategies for biomarker identification; ranging from stem cells, immune mechanisms, genomics, imaging, network science to cognition. Thereafter we emphasize key points made by contributors on affective disorders, psychosis, schizophrenia, and autism spectrum disorder. An underlying theme is how preclinical and clinical research are informing biomarker development and the importance of forward and reverse translation approaches. In considering the exploitation of biomarkers we note that there is a timely opportunity to improve clinical trial design informed by patient 'biological' and 'psychological' phenotype. This has the potential to reinvigorate drug development and clinical trials in psychiatry. In conclusion, we are poised to move from the descriptive and discovery phase to one where biomarker panels can be evaluated in real-life cohorts. This will necessitate resources for large-scale collaborative efforts worldwide. Ultimately this will lead to new interventions and personalized medicines and transform our ability to prevent illness onset and treat complex psychiatric disorders more effectively.


Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function.
Porcine Reproductive and Respiratory Syndrome (PRRS) is a panzootic infectious disease of pigs, causing major economic losses to the world-wide pig industry. PRRS manifests differently in pigs of all ages but primarily causes late-term abortions and stillbirths in sows and respiratory disease in piglets. The causative agent of the disease is the positive-strand RNA PRRS virus (PRRSV). PRRSV has a narrow host cell tropism, limited to cells of the monocyte/macrophage lineage. CD163 has been described as a fusion receptor for PRRSV, whereby the scavenger receptor cysteine-rich domain 5 (SRCR5) region was shown to be an interaction site for the virus in vitro. CD163 is expressed at high levels on the surface of macrophages, particularly in the respiratory system. Here we describe the application of CRISPR/Cas9 to pig zygotes, resulting in the generation of pigs with a deletion of Exon 7 of the CD163 gene, encoding SRCR5. Deletion of SRCR5 showed no adverse effects in pigs maintained under standard husbandry conditions with normal growth rates and complete blood counts observed. Pulmonary alveolar macrophages (PAMs) and peripheral blood monocytes (PBMCs) were isolated from the animals and assessed in vitro. Both PAMs and macrophages obtained from PBMCs by CSF1 stimulation (PMMs) show the characteristic differentiation and cell surface marker expression of macrophages of the respective origin. Expression and correct folding of the SRCR5 deletion CD163 on the surface of macrophages and biological activity of the protein as hemoglobin-haptoglobin scavenger was confirmed. Challenge of both PAMs and PMMs with PRRSV genotype 1, subtypes 1, 2, and 3 and PMMs with PRRSV genotype 2 showed complete resistance to viral infections assessed by replication. Confocal microscopy revealed the absence of replication structures in the SRCR5 CD163 deletion macrophages, indicating an inhibition of infection prior to gene expression, i.e. at entry/fusion or unpacking stages.


Selective vasodilator effects of atrial natriuretic peptide in the human placental vasculature.
To determine whether atrial natriuretic peptide (ANP) attenuates the vasoconstrictor effects of angiotensin II (AII), a thromboxane mimetic (U46619), and endothelin-1 in the human fetal-placental vasculature and to determine whether nitric oxide (NO) has a role in the vasodilator activity of ANP. Isolated placental cotyledons were dually perfused, with fetal perfusion pressure used as an index of vascular response. The effects of AII (10(-10)-10(-6) mol/L bolus injection), endothelin-1 (10(-7) mol/L bolus), and U46619 (10(-9)-10(-6) mol/L bolus or 10(-8) mol/L infusion) were established in the absence or presence of ANP (10(-8) mol/L). The role of NO as a mediator of ANP action was investigated by perfusion with n-nitro-L-arginine (NNLA, 10(-3) mol/L), an inhibitor of NO synthase. Statistical significance was determined by analysis of variance. Atrial natriuretic peptide caused significant attenuation of vasoconstrictor responses to AII, but weak attenuation of endothelin-1 and no attenuation of U46619. Use of NNLA did not affect the attenuation of AII-induced vasoconstriction by ANP. Atrial natriuretic peptide is a vasodilator of the fetal-placental vasculature constricted with AII and endothelin-1, but not with U46619. Nitric oxide does not mediate the action of ANP.


Platelet redox balance in diabetic patients with hypertension improved by n-3 fatty acids.
Patients with type 2 diabetes mellitus (T2DM) are at increased risk of developing cardiovascular disease, largely as a result of defective production of cardioprotective nitric oxide and a concomitant rise in oxidative stress. Dietary interventions that could reverse this trend would be extremely beneficial. Here we investigated whether dietary n-3 polyunsaturated fatty acid (n-3 PUFA) supplementation positively affected platelet nitroso-redox imbalance. We randomized hypertensive T2DM patients (T2DM HT; n = 22) and age-and-sex matched hypertensive study participants without diabetes (HT alone; n = 23) in a double-blind, crossover fashion to receive 8 weeks of n-3 PUFAs (1.8 g eicosapentaenoic acid and 1.5 g docosahexaenoic acid) or identical olive oil capsules (placebo), with an intervening 8-week washout period. Platelet nitrite and superoxide were measured and compared before and after treatment; 8-isoprostane was determined by ELISA and subcellular compartmentalization of the NAD(P)H oxidase subunit p47-phox examined by Western blotting. The n-3 PUFA supplementation reduced 8-isoprostane and superoxide levels in platelets from T2DM HT, but not HT alone, participants, without effect on nitrite production. This coincided with a significant decrease in p47-phox membrane localization and a similar reduction in superoxide to that achieved with apocynin. At baseline, a subcohort of T2DM HT and HT alone participants showed evidence of nitric oxide synthase (NOS)-derived superoxide production, indicating defective enzymatic activity. This was reversed significantly in T2DM HT participants after treatment, demonstrating improved NOS function. Our finding that n-3 PUFAs diminish platelet superoxide production in T2DM HT patients in vivo suggests a therapeutic role for these agents in reducing the vascular-derived oxidative stress associated with diabetes.


MiR-206, a key modulator of skeletal muscle development and disease.
MicroRNAs (miRNAs) have recently emerged as fundamental post-transcriptional regulators inhibit gene expression linked to various biological processes. MiR-206 is one of the most studied and best characterized miRNA to date, which specifically expressed in skeletal muscle. In this review, we summarized the results of studies of miR-206 with emphasis on its function in skeletal muscle development. Importantly, dysregulation of miR-206 has been linked to many disorders in skeletal muscle such as Duchenne muscular dystrophy (DMD) and amyotrophic lateral sclerosis (ALS), and circulating miR-206 has highlighted its potential as a diagnose biomarker. In addition, a mutation in the 3' untranslated region (3'-UTR) of the myostatin gene in the Texel sheep creating a target site for the miR-206 and miR-1 leads to inhibition of myostatin expression, which likely to cause the muscular hypertrophy phenotype of this breed of sheep. Therefore, miR-206 may become novel target for ameliorating skeletal muscle-related disorders and optimization of muscle quantity of domestic animals.


Genetic mapping and cloning of the gene (trmC) responsible for the synthesis of tRNA (mnm5s2U)methyltransferase in Escherichia coli K12.
The trmC gene, responsible for the formation of 5-methylaminomethyl-2-thiouridine (mnm5s2U) from 2-thiouridine, present in the first position in the anticodon of some tRNAs, has been located at 50.5 min on the Escherichia coli K12 chromosome. Results from transductional mapping suggest that the trmC gene is located counter-clockwise of aroC. A ColE1 hybrid plasmid carrying the aroC+, trmC+ and hisT+ genes was isolated, and the gene order was established, by subcloning, to be hisT-trmC-aroC. The trmC gene is located 1.9 kb from the aroC gene. Two mutations (trmC1 and trmC2) were shown to be recessive, suggesting that the trmC gene is the structural gene for the tRNA-(mnm5s2U)methyltransferase.


Persistent insomnia is a predictor of hypertension in Japanese male workers.
Insomnia is one of the most common complaints at worksites, as well as in the general population. This study aims to assess the effect of insomnia on the development of hypertension in Japanese male workers. Using the annual health examination database of a Japanese telecommunication company, eligible middle-aged male participants in the 1994 health examination were followed up until 1998 or the development of hypertension (either initiation of antihypertensive therapy or a systolic blood pressure > or = 140 mmHg and/or a diastolic blood pressure > or = 90 mmHg). The effect of difficulty initiating sleep (DIS) was assessed with a DIS dataset (n=4,794), which included non-DIS (n=4,602) and persistent-DIS (n=192) subjects. That of difficulty maintaining sleep (DMS) was assessed with a DMS dataset (n=4,443), which included non-DMS (n=4,157) and persistent-DMS (n=286) subjects. The incidence of hypertension among persistent-DIS (40.1%; 130.7 per 1,000 person-yr) was significantly higher than that among non-DIS (30.6%; 89.9 per 1,000 person-yr). The incidence of hypertension among persistent-DMS (42.3%; 136.7 per 1,000 person-yr) was significantly higher than that among non-DMS (30.7%; 90.8 per 1,000 person-yr). After adjusting for potential confounding factors (i.e. age, body mass index, smoking, alcohol drinking, and job stress), persistent complaints of DIS and DMS were significantly associated with an increased risk of hypertension (OR=1.96; 95%CI: 1.42-2.70 and OR=1.88; 95%CI: 1.45-2.45, respectively). Persistent insomnia may be a useful predictor of hypertension in Japanese male workers.