Development of Highly Potent, G-Protein Pathway Biased, Selective, and Orally Bioavailable GPR84 Agonists.
Orphan G-protein-coupled receptor 84 (GPR84) is a receptor that has been linked to cancer, inflammatory, and fibrotic diseases. We have reported DL-175 as a biased agonist at GPR84 which showed differential signaling via Gαi/cAMP and β-arrestin, but which is rapidly metabolized. Herein, we describe an optimization of DL-175 through a systematic structure-activity relationship (SAR) analysis. This reveals that the replacement of the naphthalene group improved metabolic stability and the addition of a 5-hydroxy substituent to the pyridine N-oxide group, yielding compounds 68 (OX04528) and 69 (OX04529), enhanced the potency for cAMP signaling by 3 orders of magnitude to low picomolar values. Neither compound showed detectable effects on β-arrestin recruitment up to 80 μM. Thus, the new GPR84 agonists 68 and 69 displayed excellent potency, high G-protein signaling bias, and an appropriate in vivo pharmacokinetic profile that will allow investigation of GPR84 biased agonist activity in vivo.


Risks associated with antipsychotic treatment in pregnancy: Comparative cohort studies based on electronic health records.
Limited information is available on whether antipsychotics prescribed in pregnancy are associated with increased risks of adverse outcomes. We used electronic health records from pregnant women and their children to examine risks of adverse maternal and child outcomes in three cohorts of women who: (A) received antipsychotic treatment in pregnancy (n=416) (B) discontinued antipsychotic treatment before pregnancy (n=670), and (C) had no records of antipsychotic treatment before or during pregnancy (n=318,434). Absolute and risk ratios were estimated and adjusted for health and lifestyle and concomitant medications. Caesarean section was more common in cohort A (25%) than C (18%), but non-significant after adjustment for health and lifestyle factors (Risk Ratio (adj.) 1.09 (95% CI: 0.92, 1.30). Proportion of gestational diabetes was similar in cohort A (2.6%) and B (2.7%), but lower in A than B after adjustments (RRadj: 0.43 (0.20, 0.93). Premature birth/low birthweight were more common in cohort A (10%) than B (4.3%) and C (3.9%), A versus B (RRadj: 2.04 (1.13, 3.67), A versus C (RRadj: 1.43 (0.99, 2.05). Major congenital malformations were more common in A (3.4%), than B (2.2%) and C (2%). However no significant difference was observed (A versus B: RRadj: 1.79 (0.72, 4.47) A versus C RRadj: 1.59 (0.84, 3.00)). Risks estimates were similar for women prescribed atypical and typical antipsychotics. Antipsychotic treatment in pregnancy carries limited risks of adverse pregnancy and birth outcomes once adjustments have been made for health and lifestyle factors.


Topical treatment of standardised burns with herbal remedies in model rats.
Experimental studies of burns require the use of different animal models. The aim of this work was to establish experimental model of thermal injuries and to evaluate the effects of topical agents on healing of the burn wounds. Forty female Wistar rats were randomly classified in 4 groups and isolated for 2 weeks before the onset of experiment. Animals were primarily anaesthetized with pentobarbital-sodium and then shaved (skin area of their back with diameters 5 cm x 5 cm). A round metal stamp with contact area of 5 cm2 and total weight of 100 g was heated up to 80 degrees C and then applied without additional pressure on the depilated skin of the back for 14 seconds. This procedure produced a standardized burn wound. Induced burn wounds were immediately drowned in the 4 degrees C- water for 3 s in order to maintain microcirculation. After the inducement of thermal injures, all rats were treated with 1% silver sulfadiazine cream, herbal topical preparations or were not treated at all. Burn wounds were treated twice a day until the healing completion. The result of treatment application was a significant reduction of burn wound diameters. Herbal topical preparations expressed positive therapeutic effects on the parameters of burn wounds. The efficiency of silver sulfadiazine cream in burn wound healing was significantly more expressed in comparison to healing process in control group of animals (p < or = 0,001). We conclude that herbal topical preparations efficiently caused the completion of burn wound healing process without scar formation.


Numerical analysis of the transmission efficiency of heat-drawn and chemically etched scanning near-field optical microscopes.
The scanning near-field optical microscope (SNOM) has been tested experimentally for a wide variety of applications, but, to date, there has been little work done on the numerical or analytical modeling of the optical field as it propagates throughout the SNOM probe. Therefore, the fabrication on the probes relies more on trial and error than on clear design principles. An algorithm has been developed for the study and optimization of the geometry of SNOM probes fabricated by the heat-drawn and the one-step chemically etched methods. The algorithm uses the finite-difference beam propagation method (FD-BPM) to model the field evolution throughout the SNOM structure.


Total antioxidant concentrations of breastmilk--an eye-opener to the negligent.
The balance between free radical production and antioxidant defenses in the body has important systemic and oral health implications. There is convincing evidence that breastmilk containing antioxidants is important in the prevention of diseases in infancy. This study compared the total antioxidant concentration of human breastmilk expressed at different stages of lactation, stored at various temperatures and durations. Expressed breastmilk (EBM) samples of the third, seventh and 30th day were collected from women who had term and preterm deliveries (n = 20). Another cohort of women (n = 20) was also assessed; these women were more than five months postpartum and lactating. The total antioxidant capacity (TAC) of EBM was assessed at zero hours at room temperature, at 48 hours, one week post-refrigeration (4 degrees C), and freezing (-8 degrees C) respectively using the phosphomolybdenum method. The highest antioxidant levels were found in colostrum. The TAC of EBM reduced with time and at post-refrigeration and after freezing (p < 0.0005). No significant difference in the mean TAC was observed between the EBM samples obtained from women with either term or preterm deliveries. The progressive loss of antioxidant content of EBM emphasizes the need of awareness and curtailment of the practice of storing and later use of EBM.


Mass Histology to Quantify Neurodegeneration in Drosophila.
Progressive neurodegenerative diseases like Alzheimer's disease (AD) or Parkinson's disease (PD) are an increasing threat to human health worldwide. Although mammalian models have provided important insights into the underlying mechanisms of pathogenicity, the complexity of mammalian systems together with their high costs are limiting their use. Therefore, the simple but well-established Drosophila model-system provides an alternative for investigating the molecular pathways that are affected in these diseases. Besides behavioral deficits, neurodegenerative diseases are characterized by histological phenotypes such as neuronal death and axonopathy. To quantify neuronal degeneration and to determine how it is affected by genetic and environmental factors, we use a histological approach that is based on measuring the vacuoles in adult fly brains. To minimize the effects of systematic error and to directly compare sections from control and experimental flies in one preparation, we use the 'collar' method for paraffin sections. Neurodegeneration is then assessed by measuring the size and/or number of vacuoles that have developed in the fly brain. This can either be done by focusing on a specific region of interest or by analyzing the entire brain by obtaining serial sections that span the complete head. Therefore, this method allows one to measure not only severe degeneration but also relatively mild phenotypes that are only detectable in a few sections, as occurs during normal aging.


Leaf gas exchange characteristics of three neotropical mangrove species in response to varying hydroperiod.
We determined how different hydroperiods affected leaf gas exchange characteristics of greenhouse-grown seedlings (2002) and saplings (2003) of the mangrove species Avicennia germinans (L.) Stearn., Laguncularia racemosa (L.) Gaertn. f., and Rhizophora mangle L. Hydroperiod treatments included no flooding (unflooded), intermittent flooding (intermittent), and permanent flooding (flooded). Plants in the intermittent treatment were measured under both flooded and drained states and compared separately. In the greenhouse study, plants of all species maintained different leaf areas in the contrasting hydroperiods during both years. Assimilation-light response curves indicated that the different hydroperiods had little effect on leaf gas exchange characteristics in either seedlings or saplings. However, short-term intermittent flooding for between 6 and 22 days caused a 20% reduction in maximum leaf-level carbon assimilation rate, a 51% lower light requirement to attain 50% of maximum assimilation, and a 38% higher demand from dark respiration. Although interspecific differences were evident for nearly all measured parameters in both years, there was little consistency in ranking of the interspecific responses. Species by hydroperiod interactions were significant only for sapling leaf area. In a field study, R. mangle saplings along the Shark River in the Everglades National Park either demonstrated no significant effect or slight enhancement of carbon assimilation and water-use efficiency while flooded. We obtained little evidence that contrasting hydroperiods affect leaf gas exchange characteristics of mangrove seedlings or saplings over long time intervals; however, intermittent flooding may cause short-term depressions in leaf gas exchange. The resilience of mangrove systems to flooding, as demonstrated in the permanently flooded treatments, will likely promote photosynthetic and morphological adjustment to slight hydroperiod shifts in many settings.


Androgens modulate structure and function of the suprachiasmatic nucleus brain clock.
Gonadal hormones can modulate circadian rhythms in rodents and humans, and androgen receptors are highly localized within the core region of the mouse suprachiasmatic nucleus (SCN) brain clock. Although androgens are known to modulate neural plasticity in other CNS compartments, the role of androgens and their receptors on plasticity in the SCN is unexplored. In the present study, we ask whether androgens influence the structure and function of the mouse SCN by examining the effects of gonadectomy (GDX) on the structure of the SCN circuit and its responses to light, including induction of clock genes and behavioral phase shifting. We found that after GDX, glial fibrillary acidic protein increased with concomitant decreases in the expression of the synaptic proteins synaptophysin and postsynaptic density 95. We also found that GDX exerts effects on the molecular and behavioral responses to light that are phase dependent. In late night [circadian time (CT)21], GDX increased light-induced mPer1 but not mPer2 expression compared with intact (INT) controls. In contrast, in early night (CT13.5), GDX decreased light induced mPer2 but had no effect on mPer1. At CT13.5, GDX animals also showed larger phase delays than did INT. Treatment of GDX animals with the nonaromatizable androgen dihydrotestosterone restored glial fibrillary acidic protein, postsynaptic density 95, and synaptophysin in the SCN and reinstated the INT pattern of molecular and behavioral responses to light. Together, the results reveal a role for androgens in regulating circuitry in the mouse SCN, with functional consequences for clock gene expression and behavioral responses to photic phase resetting stimuli.


Adaptive Evolution of Eel Fluorescent Proteins from Fatty Acid Binding Proteins Produces Bright Fluorescence in the Marine Environment.
We report the identification and characterization of two new members of a family of bilirubin-inducible fluorescent proteins (FPs) from marine chlopsid eels and demonstrate a key region of the sequence that serves as an evolutionary switch from non-fluorescent to fluorescent fatty acid-binding proteins (FABPs). Using transcriptomic analysis of two species of brightly fluorescent Kaupichthys eels (Kaupichthys hyoproroides and Kaupichthys n. sp.), two new FPs were identified, cloned and characterized (Chlopsid FP I and Chlopsid FP II). We then performed phylogenetic analysis on 210 FABPs, spanning 16 vertebrate orders, and including 163 vertebrate taxa. We show that the fluorescent FPs diverged as a protein family and are the sister group to brain FABPs. Our results indicate that the evolution of this family involved at least three gene duplication events. We show that fluorescent FABPs possess a unique, conserved tripeptide Gly-Pro-Pro sequence motif, which is not found in non-fluorescent fatty acid binding proteins. This motif arose from a duplication event of the FABP brain isoforms and was under strong purifying selection, leading to the classification of this new FP family. Residues adjacent to the motif are under strong positive selection, suggesting a further refinement of the eel protein's fluorescent properties. We present a phylogenetic reconstruction of this emerging FP family and describe additional fluorescent FABP members from groups of distantly related eels. The elucidation of this class of fish FPs with diverse properties provides new templates for the development of protein-based fluorescent tools. The evolutionary adaptation from fatty acid-binding proteins to fluorescent fatty acid-binding proteins raises intrigue as to the functional role of bright green fluorescence in this cryptic genus of reclusive eels that inhabit a blue, nearly monochromatic, marine environment.


Angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers reduced dementia risk in patients with diabetes mellitus and hypertension.
The effects of angiotensin-converting enzyme inhibitors (ACEI) and angiotensin II receptor blockers (ARB) on dementia risk in patients with type 2 diabetes mellitus (DM) and hypertension remain unknown. We investigated the effects of ACEIs and ARBs on dementia risk in patients with type 2 DM and hypertension. We conducted a cohort study by using the Taiwan National Health Insurance Research Database. We included 2377 patients receiving ACEIs and 1780 patients receiving ARBs in the ACEI and ARB cohorts, respectively. We included a comparable number of patients not receiving ACEIs and ARBs as controls in the non-ACEI and non-ARB cohorts through propensity score matching. The effect of ACEIs and ARBs on dementia risk was estimated through multivariate Cox proportional hazard regression after adjustment for several confounding factors. During the 12-year follow-up period, compared with the non-ACEI cohort, all-cause dementia risk decreased by 26% in the ACEI cohort [hazard ratio (HR)=0.74, 95% confidence interval (CI)=0.56-0.96]. The all-cause dementia risk was nearly 40% lower in the ARB cohort than in the non-ARB cohort (HR=0.60, 95% CI=0.37-0.97). These drugs prevented the occurrence of vascular dementia (VD), however, this effect was nonsignificant for Alzheimer's dementia (AD). Treatment duration- and dosage-related protection effects on dementia occurrence were observed. ACEIs and ARBs may effectively prevent all-cause dementia, particularly VD, in patients with type 2 DM and hypertension. Moreover, compared with ACEIs, ARBs appear to be more advantageous in dementia prevention.


A 'mixed reality' simulator concept for future Medical Emergency Response Team training.
The UK Defence Medical Service's Pre-Hospital Emergency Care (PHEC) capability includes rapid-deployment Medical Emergency Response Teams (MERTs) comprising tri-service trauma consultants, paramedics and specialised nurses, all of whom are qualified to administer emergency care under extreme conditions to improve the survival prospects of combat casualties. The pre-deployment training of MERT personnel is designed to foster individual knowledge, skills and abilities in PHEC and in small team performance and cohesion in 'mission-specific' contexts. Until now, the provision of airborne pre-deployment MERT training had been dependent on either the availability of an operational aircraft (eg, the CH-47 Chinook helicopter) or access to one of only two ground-based facsimiles of the Chinook's rear cargo/passenger cabin. Although MERT training has high priority, there will always be competition with other military taskings for access to helicopter assets (and for other platforms in other branches of the Armed Forces). This paper describes the development of an inexpensive, reconfigurable and transportable MERT training concept based on 'mixed reality' technologies-in effect the 'blending' of real-world objects of training relevance with virtual reality reconstructions of operational contexts.


Melanoma cell adhesion molecule is the driving force behind the dissemination of melanoma upon S100A8/A9 binding in the original skin lesion.
Since metastasis accounts for the majority of cancer-associated deaths, studies on the mechanisms of metastasis are needed to establish innovative strategies for cancer treatment. We previously reported that melanoma cell adhesion molecule (MCAM) functions as a critical receptor for S100A8/A9, and binding of S100A8/A9 to MCAM results in the migration of melanoma cells to lung tissue. However, the critical role of MCAM in the original melanoma skin lesion is still not clear. In this study, we aimed to determine the importance of the S100A8/A9-MCAM axis in melanoma dissemination in a skin lesion as a critical early step for metastasis. Mechanistic studies revealed the downstream signaling of MCAM that signaled the induction of metastasis. S100A8/A9-MCAM binding activates mitogen-activated protein kinase kinase kinase 8 (MAP3K8), also termed TPL2, leading to strong activation of the transcription factor ETV4 and subsequent induction of matrix metalloproteinase-25 (MMP25), and finally to induction of melanoma lung tropic metastasis. Collectively, our results demonstrate a crucial role of the S100A8/A9-MCAM signaling axis in metastatic onset of melanoma cells and indicate that strategies targeting the identified pathway may be useful for the establishment of innovative anti-cancer therapies.


Use of serologic markers as a screening tool in inflammatory bowel disease compared with elevated erythrocyte sedimentation rate and anemia.
The purpose of this work was to evaluate the use of serologic testing as a screening test for inflammatory bowel disease compared with erythrocyte sedimentation rate and hemoglobin in a referred patient population with suspected inflammatory bowel disease. A retrospective study was performed, reviewing medical charts of patients who had inflammatory bowel disease serology performed at Prometheus Laboratories from September 2002 to September 2004. Patients were divided into 4 categories: ulcerative colitis, Crohn disease, indeterminate colitis, and noninflammatory bowel disease groups. Patients were categorized based on clinical evaluation by board-certified pediatric gastroenterologists. A total of 227 patients seen at the Lucile Packard Children's Hospital Gastroenterology Clinic had inflammatory bowel disease serology performed at or before the time of diagnosis. Seventeen charts were excluded secondary to inadequate information. Forty children were found to have inflammatory bowel disease, a prevalence of 19%. Overall, serologic testing for inflammatory bowel disease had 60% sensitivity and 92% specificity. A positive laboratory test for anemia or an elevated erythrocyte sedimentation rate had 83% sensitivity, whereas the combination of anemia and elevated erythrocyte sedimentation rate had 96% specificity. The positive predictive value of serologic testing was 60% compared with 79% in patients with anemia and elevated erythrocyte sedimentation rate. The positive predictive value of serologic testing in the subgroup of subjects without rectal bleeding (139 subjects) was only 35% compared with 60% using routine tests. Almost one third of all positive serologic tests were in patients with no demonstrable inflammatory bowel disease. As a pediatric inflammatory bowel disease screening strategy for the general pediatrician or gastroenterologist, the measurement of the combination of erythrocyte sedimentation rate and hemoglobin has a higher positive predictive value and is more sensitive, more specific, and less costly than commercial serologic testing.


Understanding efflux in Gram-negative bacteria: opportunities for drug discovery.
Bacteria evolved an arsenal of mechanisms to deal with toxic compounds and metabolic stresses, including antimicrobial agents. Efflux pumps are major players in the multidrug resistance of Gram-negative bacteria and pose major hurdles in the drug discovery process. However, recent advances in our understanding of efflux in these bacteria provide opportunities and assets for drug discovery. This review provides an overview of drug efflux in Gram-negative bacteria and its role in antimicrobial resistance, stress responses and other biological processes such as biofilm formation, and virulence. The discussion includes comments on the significance of synergy between a low-permeability outer membrane and efflux, notably the role of porins and lipopolysaccharide. The author then summarizes efforts aimed at inhibiting efflux pumps as a means to extend the utility of clinically useful antibiotics. This includes highlights of identification and characterization of small molecule efflux pump inhibitors (EPIs) from natural and synthetic sources, as well as novel strategies such as gene silencing and inhibitory antibodies. Options for treating infections caused by multidrug-resistant bacteria are limited. Given the attractiveness of the therapeutic potential of efflux pump inhibition, further studies exploring novel strategies to interfere with efflux pump expression and function are warranted. This includes rational EPI design facilitated by pump structure information, exploitation of genetically defined efflux-proficient and efflux-compromised strain panels and non-traditional approaches such as pump inhibition by gene silencing, antibodies and perhaps even phage.


Heuristic cognitive diagnosis when the Q-matrix is unknown.
Cognitive diagnosis models of educational test performance rely on a binary Q-matrix that specifies the associations between individual test items and the cognitive attributes (skills) required to answer those items correctly. Current methods for fitting cognitive diagnosis models to educational test data and assigning examinees to proficiency classes are based on parametric estimation methods such as expectation maximization (EM) and Markov chain Monte Carlo (MCMC) that frequently encounter difficulties in practical applications. In response to these difficulties, non-parametric classification techniques (cluster analysis) have been proposed as heuristic alternatives to parametric procedures. These non-parametric classification techniques first aggregate each examinee's test item scores into a profile of attribute sum scores, which then serve as the basis for clustering examinees into proficiency classes. Like the parametric procedures, the non-parametric classification techniques require that the Q-matrix underlying a given test be known. Unfortunately, in practice, the Q-matrix for most tests is not known and must be estimated to specify the associations between items and attributes, risking a misspecified Q-matrix that may then result in the incorrect classification of examinees. This paper demonstrates that clustering examinees into proficiency classes based on their item scores rather than on their attribute sum-score profiles does not require knowledge of the Q-matrix, and results in a more accurate classification of examinees.


RIG-I-like receptors: sensing and responding to RNA virus infection.
Viral and microbial pathogens contain specific motifs or pathogen-associated molecular patterns (PAMPs) that are recognized by cell surface- and endosome-associated Toll-like receptors (TLRs). RNA virus infection is also detected through TLR-independent mechanisms. Early viral replicative intermediates are detected by two recently characterized cystolic viral RNA receptors-RIG-I and MDA-5. Both are DExDH/box RNA helicases, and RIG-I specifically recognizes 5'-triphosphate containing viral RNA and transmits signals that induce type I interferon-mediated host immunity against virus infection. In this review, we will focus on RIG-I-like receptor (RLR) signal transduction and the regulatory mechanisms - ubiquitination, deubiquitination, ISGylation - underlying this important host response.


Digital dermatitis treponemes associated with a severe foot disease in dairy goats.
A UK dairy goat herd was assessed after reports of a severe lameness problem of unknown aetiology. A lameness prevalence estimate was produced and individual clinical examination of 15 randomly selected lame goats was performed. Fifteen animals had foot lesions closely resembling contagious ovine digital dermatitis (CODD) in sheep. Eight of the goats examined presented with typical CODD lesions and seven showed what appeared to be a more severe CODD with under-running of the sole. Ten biopsy samples were obtained from the foot lesions and tested by PCR for the three previously isolated digital dermatitis (DD) Treponema phylogroups and culture of treponemes was attempted. Ninety per cent of the biopsy samples were positive for Treponema medium/Treponema vincentii-like spirochaetes and Treponema phagedenis-like DD spirochaetes and 80per cent were positive for Treponema pedis. Spirochaetes were successfully isolated from 50 per cent of lesion samples. Three isolates were identified as belonging to the T. phagedenis-like spirochaetes and two were identified as T. pedis. The frequent isolation of similar treponemes to those isolated from bovine digital dermatitis and CODD lesions and the identification of these DD-associated phylotypes in the vast majority of lesions support the hypothesis that this novel foot condition is associated with infection by DD treponemes, and given the similarities to CODD, it suggests a causal role.


Expression of a platelet-derived growth factor-like protein in simian sarcoma virus transformed cells.
The near identity of the partial amino acid sequence of human platelet-derived growth factor (PDGF) and that predicted for p28sis, the putative transforming protein of the simian sarcoma virus (SSV), suggests expression of a growth factor activity may be central for transformation by SSV. It is now reported that SSV-transformed cells but not control cells contain a growth factor activity that is identical to PDGF in immunoassay, in mitogenic dose response, and in specific mitogenic activity. The protein immunoprecipitated by antiserum to human PDGF has an apparent molecular weight of 20,000, identical to that of p20sis, the putative intracellular degradation product of p28sis. The results support the concept that expression of a PDGF-like molecule, which appears to be the product of the viral-sis gene, is responsible for the abnormal regulation of growth is SSV-transformed cells.


Radionuclides in wastewater treatment plants: monitoring of Sicilian plants.
Three Sicilian wastewater treatment plants were monitored to assess the occurrence and the behaviour of radionuclides. Two sampling campaigns (screening and long-term) were carried out during which liquid and solid samples have been analysed. It was found that ¹³¹I mostly occurred in the samples analysed during the screening campaign (43% of the analysed samples contained ¹³¹I). High ¹³¹I specific activity was found in the mixed liquor, recycled sludge and dehydrated sludge samples. This finding was mainly due to the tendency of (131)I to be associated with solid particles. During the long-term sampling campaign an influence of the sludge retention time (SRT) on the ¹³¹I behaviour was found. Increasing the SRT and consequently decreasing the fraction of active organic biomass inside the system, the specific activity of ¹³¹I decreases.


Isolation and characterization of 10 MHC Class I-associated microsatellite loci in tammar wallaby (Macropus eugenii).
The major histocompatibility complex (MHC) contain genes which play a key role in immune response and mate choice, and are therefore of functional importance to molecular ecologists. Here we describe the design of 10 MHC Class I-associated microsatellite loci from the tammar wallaby. All 10 loci are highly polymorphic, with the expected heterozygosity ranging from 0.547 to 0.919. Six loci successfully cross-amplify in other macropodid species. These microsatellites will serve as useful tools for studying the level of MHC diversity, the impact of selection on genetic variation and the unique structure of the tammar wallaby MHC.


Hypertensive coronary microvascular dysfunction: a subclinical marker of end organ damage and heart failure.
Hypertension is a well-established heart failure (HF) risk factor, especially in the context of adverse left ventricular (LV) remodelling. We aimed to use myocardial flow reserve (MFR) and global longitudinal strain (GLS), markers of subclinical microvascular and myocardial dysfunction, to refine hypertensive HF risk assessment. Consecutive patients undergoing symptom-prompted stress cardiac positron emission tomography (PET)-computed tomography and transthoracic echocardiogram within 90 days without reduced left ventricular ejection fraction (<40%) or flow-limiting coronary artery disease (summed stress score ≥ 3) were included. Global MFR was quantified by PET, and echocardiograms were retrospectively analysed for cardiac structure and function. Patients were followed over a median 8.75 (Q1-3 4.56-10.04) years for HF hospitalization and a composite of death, HF hospitalization, MI, or stroke. Of 194 patients, 155 had adaptive LV remodelling while 39 had maladaptive remodelling, which was associated with lower MFR and impaired GLS. Across the remodelling spectrum, diastolic parameters, GLS, and N-terminal pro-B-type natriuretic peptide were independently associated with MFR. Maladaptive LV remodelling was associated with increased adjusted incidence of HF hospitalization and death. Importantly, the combination of abnormal MFR and GLS was associated with a higher rate of HF hospitalization compared to normal MFR and GLS [adjusted hazard ratio (HR) 3.21, 95% confidence interval (CI) 1.09-9.45, P = 0.034), including in the adaptive remodelling subset (adjusted HR 3.93, 95% CI 1.14-13.56, P = 0.030). We have demonstrated important associations between coronary microvascular dysfunction and myocardial mechanics that refine disease characterization and HF risk assessment of patients with hypertension based on subclinical target organ injury.


REVIEW. Melatonin and the thyroid gland.
This review briefly summarizes the published data on relationships observed between melatonin - the main pineal hormone, and the thyroid gland. The prevailing part of the survey is devoted to thyroid growth processes and function. A large experimental evidence exists suggesting the inhibitory action of melatonin on thyroid growth and function; this effect has been revealed by using different experimental models: by chronic and short-term melatonin administration in vivo, by light restriction, which is known to increase the activity of the pineal gland, by pinealectomy, etc., as well as by employing the in vitro conditions. Thus, much information has been accumulated, indicating - in experimental conditions - a mutual relationship between the pineal gland and the thyroid. The confirmation of these relations in clinical studies in humans meets numerous difficulties, resulting - among others - from the fact that, nowadays, human beings, as well as certain animal species, used in experimental studies, have been living far away from their natural and original habitat. It makes almost impossible to compare the results obtained in particular studies performed in different species, on the pineal-thyroid interrelationship.


Remarkable affinity and selectivity for Cs+ and uranyl (UO22+) binding to the manganese site of the apo-water oxidation complex of photosystem II.
The size and charge density requirements for metal ion binding to the high-affinity Mn2+ site of the apo-water oxidizing complex (WOC) of spinach photosystem II (PSII) were studied by comparing the relative binding affinities of alkali metal cations, divalent metals (Mg2+, Ca2+, Mn2+, Sr2+), and the oxo-cation UO22+. Cation binding to the apo-WOC-PSII protein was measured by: (1) inhibition of the rate and yield of photoactivation, the light-induced recovery of O2 evolution by assembly of the functional Mn4Ca1Clx, core from its constituent inorganic cofactors (Mn2+, Ca2+, and Cl-); and by (2) inhibition of the PSII-mediated light-induced electron transfer from Mn2+ to an electron acceptor (DCIP). Together, these methods enable discrimination between inhibition at the high- and low-affinity Mn2+ sites and the Ca2+ site of the apo-WOC-PSII. Unexpectedly strong binding of large alkali cations (Cs+ >> Rb+ > K+ > Na+ > Li+) was found to smoothly correlate with decreasing cation charge density, exhibiting one of the largest Cs+/Li+ selectivities (>/=5000) for any known chelator. Both photoactivation and electron-transfer measurements at selected Mn2+ and Ca2+ concentrations reveal that Cs+ binds to the high-affinity Mn2+ site with a slightly greater affinity (2-3-fold at pH 6.0) than Mn2+, while binding about 10(4)-fold more weakly to the Ca2+-specific site required for reassembly of functional O2 evolving centers. In contrast to Cs+, divalent cations larger than Mn2+ bind considerably more weakly to the high-affinity Mn2+ site (Mn2+ >> Ca2+ > Sr2+). Their affinities correlate with the hydrolysis constant for formation of the metal hydroxide by hydrolysis of water: Me2+aq --> [MeOH]+aq + H+aq. Along with the strong stimulation of the rate of photoactivation by alkaline pH, these metal cation trends support the interpretation that [MnOH]+ is the active species that forms upon binding of Mn2+aq to apo-WOC. Further support for this interpretation is found by the unusually strong inhibition of Mn2+ photooxidation by the linear uranyl cation (UO22+). The intrinsic binding constant for [MnOH]+ to apo-WOC was determined using a thermodynamic cycle to be K = 4.0 x 10(15) M-1 (at pH 6.0), consistent with a high-affinity, preorganized, multidentate coordination site. We propose that the selectivity for binding [MnOH]+, a linear low charge-density monocation, vs symmetrical Me2+ dications is functionally important for assembly of the WOC by enabling: (1) discrimination against higher charge density alkaline earth cations (Mg2+ and Ca2+) and smaller alkali metal cations (Na+ and K+) that are present in considerably greater abundance in vivo, and thus would suppress photoactivation; and (2) higher affinity binding of the one Ca2+ ion or the remaining three Mn2+ ions via coordination to form mu-hydroxo-bridged intermediates, apo-WOC-[Mn(mu-OH)2Mn]3+ or apo-WOC-[Mn(mu-OH)Ca]3+, during subsequent assembly steps of the native Mn4Ca1Clx core. In contrast to more acidic Me2+ divalent ion inhibitors of the high-affinity Mn2+ site, like Ca2+ and Sr2+, Cs+ does not accelerate the decay of the first light-induced intermediate, IM1, formed during photoactivation (attributed to apo-WOC-[Mn(OH)2]+). The inability of Cs+ to promote decay of IM1, despite having comparable affinity as Mn2+, is consistent with its considerably weaker Lewis acidity, resulting in the reprotonation of IM1 by water becoming the rate-limiting step for decay prior to displacement of Mn2+. All four different lines of evidence provide a self-consistent picture indicating that the initial step in assembly of the WOC involves high-affinity binding of [MnOH]+.


Effect of magnetic seizure therapy on regional brain glucose metabolism in major depression.
Currently electroconvulsive therapy (ECT) is one of the only available therapies for treatment resistant depression (TRD). While effective, ECT is complicated by side effects, including cognitive impairment. One promising potential alternative is magnetic seizure therapy (MST). To date, no research has explored the effects of 100Hz MST on brain activity or the brain changes associated with response to treatment. Therefore the aim of this study was to determine the effects of a treatment course of 100Hz MST on regional brain glucose metabolism. Ten patients with treatment resistant depression underwent positron emission tomography with fluorodeoxyglucose before and after a treatment course of MST. Changes in the relative metabolic rate of a priori brain regions were investigated. Areas of increased relative metabolism after treatment were seen in the basal ganglia, orbitofrontal cortex, medial frontal cortex and dorsolateral prefrontal cortex. A secondary analysis showed trend-level differential findings in brain activation between responders and non-responders, namely in the ventral anterior cingulate. These results primarily indicate that MST is affecting regions consistent with the limbic-cortical dysregulation model of depression. Exploratory analysis indicated some differential findings in brain activation between responders and non-responders were also evident; however, the small sample size precludes any firm conclusions.


Virome characterisation from Guthrie cards in children who later developed acute lymphoblastic leukaemia.
Some childhood acute lymphoblastic leukaemias (ALL) can be traced back to a prenatal origin, where a virus infection could be involved in the first pre-leukaemic clone development. The DNA virome of 95 children who later developed ALL was characterised from neonatal blood spots (NBS) using unbiased next-generation sequencing (NGS) and compared with the virome of 95 non-ALL controls. DNA was individually extracted from the ALL-patients and controls, pooled, randomly amplified and sequenced using the Illumina MiSeq Sequencing System. Virus-like sequences identified in both groups mapped to human endogenous retroviruses and propionibacterium phage, considered a part of the normal microbial flora. Potential pathogens human herpesvirus type 6 (HHV-6) and parvovirus B19 were also identified, but only few samples in both ALL and controls tested positive by PCR follow-up. Unbiased NGS was employed to search for DNA from potential infectious agents in neonatal samples of children who later developed ALL. Although several viral candidates were identified in the NBS samples, further investigation by PCR suggested that these viruses did not have a major role in ALL development.


Growth of Pasteurella haemolytica and production of its leukotoxin in semi-defined media.
To develop semi-defined media that support growth of the bovine pathogen, Pasteurella haemolytica, and use them to examine production of leukotoxin and an arginine-binding protein by this organism. 10 P haemolytica A1 strains and 1 P multocida strain. Bacterial strains were cultivated at 37 C in media containing various amino acids, carbon sources, vitamins, and cofactors, and absorbance (OD600) was measured. Leukotoxin and arginine-binding protein production were assessed by immunoblot analysis. Optimal growth required supplementation with 0.1% fetal bovine serum, gelatin, or purified bovine serum albumin. Calcium pantothenate and thiamine were essential for growth, and a variety of carbon sources could be utilized. In the complete medium, 15 amino acids were included; however, in the minimal medium, no amino acids were required. All strains (except P multocida) grew in the complete medium and 7 grew well in the minimal medium. Leukotoxin was not produced when amino acids were limiting, but could be enhanced by addition of 0.2% NH4SO4. Production of the arginine-binding protein was not affected by nitrogen availability or by presence of L-arginine. Two media that support good growth of P haemolytica strains were developed. The minimal medium is simple to prepare and manipulate and its use revealed a potential role of nitrogen availability in the regulation of leukotoxin expression. Creation of these media will permit continued studies of the response of P haemolytica to environmental conditions that may mimic those encountered in the bovine respiratory tract during shipping.


Claudin-10 is required for relay of left-right patterning cues from Hensen's node to the lateral plate mesoderm.
Species-specific symmetry-breaking events at the left-right organizer (LRO) drive an evolutionarily-conserved cascade of gene expression in the lateral plate mesoderm that is required for the asymmetric positioning of organs within the body cavity. The mechanisms underlying the transfer of the left and right laterality information from the LRO to the lateral plate mesoderm are poorly understood. Here, we investigate the role of Claudin-10, a tight junction protein, in facilitating the transfer of left-right identity from the LRO to the lateral plate mesoderm. Claudin-10 is asymmetrically expressed on the right side of the chick LRO, Hensen's node. Gain- and loss-of-function studies demonstrated that right-sided expression of Claudin-10 is essential for normal rightward heart tube looping, the first morphological asymmetry during organogenesis. Manipulation of Claudin-10 expression did not perturb asymmetric gene expression at Hensen's node, but did disrupt asymmetric gene expression in the lateral plate mesoderm. Bilateral expression of Claudin-10 at Hensen's node prevented expression of Nodal, Lefty-2 and Pitx2c in the left lateral plate mesoderm, while morpholino knockdown of Claudin-10 inhibited expression of Snail1 in the right lateral plate mesoderm. We also determined that amino acids that are predicted to affect ion selectivity and protein interactions that bridge Claudin-10 to the actin cytoskeleton were essential for its left-right patterning function. Collectively, our data demonstrate a novel role for Claudin-10 during the transmission of laterality information from Hensen's node to both the left and right sides of the embryo and demonstrate that tight junctions have a critical role during the relay of left-right patterning cues from Hensen's node to the lateral plate mesoderm.


Development of a clinical prediction rule to diagnose Pneumocystis jirovecii pneumonia in the World Health Organization's algorithm for seriously ill HIV-infected patients.
The World Health Organization (WHO) algorithm for the diagnosis of tuberculosis in seriously ill HIV-infected patients recommends that treatment for Pneumocystis jirovecii pneumonia (PJP) should be considered without giving clear guidance on selecting patients for empiric PJP therapy. PJP is a common cause of hospitalisation in HIV-infected patients in resource-poor settings where diagnostic facilities are limited. We developed clinical prediction rules for PJP in a prospective cohort of HIV-infected inpatients with WHO danger signs and cough of any duration. The reference standard for PJP was > 1000 copies/mL of P. jirovecii DNA on real-time sputum polymerase chain reaction (PCR). Four potentially predictive variables were selected for regression models: dyspnoea, chest X-ray, haemoglobin and oxygen saturation. Respiratory rate was explored as a replacement for oxygen saturation as pulse oximetry is not always available in resource-poor settings. We enrolled 500 participants. After imputation for missing values, there were 56 PJP outcome events. Dyspnoea was not independently associated with PJP. Oxygen saturation and respiratory rate were inversely correlated. Two clinical prediction rules were developed: chest X-ray possible/likely PJP, haemoglobin ≥ 9 g/dL and either oxygen saturation < 94% or respiratory rate. The area under the receiver operating characteristic curve of the clinical prediction rule models was 0.761 (95% CI 0.683-0.840) for the respiratory rate model and 0.797 (95% CI 0.725-0.868) for the oxygen saturation model. Both models had zero probability for PJP for scores of zero, and positive likelihood ratios exceeded 10 for high scores. We developed simple clinical prediction rules for PJP, which, if externally validated, could assist decision-making in the WHO seriously ill algorithm.


Simple, efficient estimators of treatment effects in randomized trials using generalized linear models to leverage baseline variables.
Models, such as logistic regression and Poisson regression models, are often used to estimate treatment effects in randomized trials. These models leverage information in variables collected before randomization, in order to obtain more precise estimates of treatment effects. However, there is the danger that model misspecification will lead to bias. We show that certain easy to compute, model-based estimators are asymptotically unbiased even when the working model used is arbitrarily misspecified. Furthermore, these estimators are locally efficient. As a special case of our main result, we consider a simple Poisson working model containing only main terms; in this case, we prove the maximum likelihood estimate of the coefficient corresponding to the treatment variable is an asymptotically unbiased estimator of the marginal log rate ratio, even when the working model is arbitrarily misspecified. This is the log-linear analog of ANCOVA for linear models. Our results demonstrate one application of targeted maximum likelihood estimation.


Interpreting small quantities of DNA: the hierarchy of propositions and the use of Bayesian networks.
The dramatic increase in the sensitivity of DNA profiling systems that has occurred over recent years has led to the need to address a wider range of interpretational problems in forensic science. The issues surrounding questions of the kind "whose DNA is this?" have been the subject of considerable controversy but now it is clear that the emphasis is shifting to questions of the kind "how did this DNA get here?" Such issues are discussed in this paper and new insights are provided by two particular recent developments. First, the notion of the "hierarchy of propositions" that has arisen from a project called Case Assessment and Interpretation (CAI) that has been running in the British Forensic Science Service (FSS). Second, a technique for drawing inferences in the face of many interacting considerations, known as "Bayesian networks"--or "Bayes' nets" for short--that has been the subject of an earlier paper in this journal (1). The discussion is carried out by means of case studies, based on actual cases. It is clear that, whereas the inference in relation to the source of the DNA in a crime sample might be overwhelmingly strong, the inference in relation to the propositions that a jury must consider relating to the identity of the actual offender may be much more tentative.


Single n+-i-n+ InP nanowires for highly sensitive terahertz detection.
Developing single-nanowire terahertz (THz) electronics and employing them as sub-wavelength components for highly-integrated THz time-domain spectroscopy (THz-TDS) applications is a promising approach to achieve future low-cost, highly integrable and high-resolution THz tools, which are desirable in many areas spanning from security, industry, environmental monitoring and medical diagnostics to fundamental science. In this work, we present the design and growth of n+-i-n+ InP nanowires. The axial doping profile of the n+-i-n+ InP nanowires has been calibrated and characterized using combined optical and electrical approaches to achieve nanowire devices with low contact resistances, on which the highly-sensitive InP single-nanowire photoconductive THz detectors have been demonstrated. While the n+-i-n+ InP nanowire detector has a only pA-level response current, it has a 2.5 times improved signal-to-noise ratio compared with the undoped InP nanowire detector and is comparable to traditional bulk THz detectors. This performance indicates a promising path to nanowire-based THz electronics for future commercial applications.


Older LGBT+ health inequalities in the UK: setting a research agenda.
Lesbian, gay, bisexual and trans+ (LGBT+) people report poorer health than the general population and worse experiences of healthcare particularly cancer, palliative/end-of-life, dementia and mental health provision. This is attributable to: (a) social inequalities, including 'minority stress'; (b) associated health-risk behaviours (eg, smoking, excessive drug/alcohol use, obesity); (c) loneliness and isolation, affecting physical/mental health and mortality; (d) anticipated/experienced discrimination and (e) inadequate understandings of needs among healthcare providers. Older LGBT+ people are particularly affected, due to the effects of both cumulative disadvantage and ageing. There is a need for greater and more robust research data to support growing international and national government initiatives aimed at addressing these health inequalities. We identify seven key research strategies: (1) Production of large data sets; (2) Comparative data collection; (3) Addressing diversity and intersectionality among LGBT+ older people; (4) Investigation of healthcare services' capacity to deliver LGBT+ affirmative healthcare and associated education and training needs; (5) Identification of effective health promotion and/or treatment interventions for older LGBT+ people, and subgroups within this umbrella category; (6) Development of an (older) LGBT+ health equity model; (7) Utilisation of social justice concepts to ensure meaningful, change-orientated data production which will inform and support government policy, health promotion and healthcare interventions.


Psychological variables potentially implicated in opioid-related mortality as observed in clinical practice.
Opioid-related deaths in the United States have become a public health problem, with accidental and unintended overdoses being especially troubling. Screening for psychological risk factors is an important first step in safeguarding against nonadherence practices and identifying patients who may be vulnerable to the risks associated with opioid therapy. Validated screening instruments can aid in this attempt as a complementary tool to clinicians' assessments. A structured screening is imperative as part of an assessment, as clinician judgment is not the most reliable method of identifying nonadherence. As a complement to formal screening, we present for discussion and possible future study certain psychological variables observed during years of clinical practice that may be linked to medication nonadherence and accidental overdose. These variables include catastrophizing, fear, impulsivity, attention deficit disorders, existential distress, and certain personality disorders. In our experience, chronic pain patients with dual diagnoses may become "chemical copers" as a way of coping with their negative emotion. For these patients, times of stress could lead to accidental overdose. Behavioral, cognitive-behavioral (acceptance and commitment, dialectical behavior), existential (meaning-centered, dignity), and psychotropic therapies have been effective in treating these high-risk comorbidities, while managing expectations of pain relief appears key to preventing accidental overdose.


Factors influencing QTc interval prolongation during kidney transplantation.
During renal transplantation, factors causing disturbances of the repolarization process, defined by the QT interval, may lead to ventricular arrhythmias. The aim of this study was to determine a relationship between QT interval prolongation and metabolic homoeostasis, and durations of cold ischemia time (CIT) and warm ischemia time (WIT) during renal transplantation. From among 198 consecutive renal transplant patients, 68 (mean age 45 ± 12 years) were included in a prospective observational pilot study. Prior to the procedure, arterial blood gas analysis was performed, and digital Holter ECG monitoring was applied and continued for 12 hours. Subsequent arterial blood gas analysis was performed 30 minutes after graft reperfusion. QTc changed dynamically and significantly during the perioperative period. Ventricular arrhythmias were observed only during graft reperfusion. Recordings showed that 33 out of 68 patients had ventricular extrasystoles, and non-sustained ventricular tachycardia was observed in 2 patients. No patients presented with hemodynamic instability. There was no statistical correlation between CIT or WIT and the difference (delta) between the final and initial values of the pH, potassium and lactate levels, QTc range, maximal QTc or QTc measured at the predefined time points. The renal transplantation procedure carries a high risk of ventricular repolarization period disturbances that can lead to life-threatening tachyarrhythmias despite optimal hemodynamic or metabolic status and independent of CIT and WIT.


Human papillomavirus infection and p53 protein expression in vulvar intraepithelial neoplasia and invasive squamous cell carcinoma.
The etiopathogenesis of vulvar intraepithelial neoplasia (VIN III) and invasive squamous cell carcinoma are largely unknown. Since there are few studies on Brazilian patients, our purpose was to determine the frequency of human papillomavirus (HPV) infection and the expression of p53 in these lesions, and associate them with other factors such as age, morphological subtypes, multicentric and multifocal disease. Thirty-eight cases of VIN III, nine of superficially invasive carcinoma, and 55 of invasive vulvar carcinoma were retrospectively evaluated from 1983 to 1995 for the presence of HPV by immunohistochemistry and in situ hybridization, and for p53 protein expression by immunohistochemistry on paraffin sections. All cases for whom material (slides and paraffin blocks) and clinical data were available were included. HPV and p53 were detected in 57.9 and 21.1% of the VIN III lesions, 33.3 and 66.7% of superficially invasive carcinomas, and 7.3 and 58.2% of invasive squamous cell carcinomas, respectively. HPV infection was associated with younger age in the VIN III and invasive carcinoma groups. In the latter, HPV infection was associated with the basaloid variant. p53 expression rate was higher in superficially invasive and invasive lesions and was not related to HPV infection. Our findings are similar to others and support the hypothesis that there are two separate entities of the disease, one associated with HPV and the other unrelated, with p53 inactivation possibly being implicated in some of the cases.


Operant controlled neural event: formal and systematic approach to electrical coding of behavior in brain.
Traditional studies of electrophysiological correlates of behavior contain inherent high variability resulting from the arbitrary choice of behaviors, brain locations, and wave parameters. The operant control of neural events is a formal and systematic approach to the study of prespecified parameters and components of brain activity as they encode behaviors. Two studies in which the electrical activity of brain was the criterion for reinforcement demonstrate the acquisition, under such operant control, of two mutually exclusive behaviors or states which selectively alter evoked potential components.


Development of the self-concept during adolescence.
Adolescence is a period of life in which the sense of 'self' changes profoundly. Here, we review recent behavioural and neuroimaging studies on adolescent development of the self-concept. These studies have shown that adolescence is an important developmental period for the self and its supporting neural structures. Recent neuroimaging research has demonstrated that activity in brain regions associated with self-processing, including the medial prefrontal cortex, changes between early adolescence and adulthood. These studies indicate that neurocognitive development might contribute to behavioural phenomena characteristic of adolescence, such as heightened self-consciousness and susceptibility to peer influence. We attempt to integrate this recent neurocognitive research on adolescence with findings from developmental and social psychology.


What predicts nurse faculty members' intent to stay in the academic organization? A structural equation model of a national survey of nursing faculty.
To investigate the relations among several factors regarding the academic context within a nationally representative sample of U.S. nursing faculty. Correlational design using structural equation modeling to explore the predictive nature of several factors related to the academic organization and the work life of nursing faculty. A survey was used to evaluate several aspects of the work life of U.S. nursing faculty members. Nursing faculty members in academic organizations across the U.S. serving at either CCNE- or NLNAC-accredited institutions of higher education. Standard confirmatory factor analysis was used to assess the validity of a proposed measurement model, and structural equation modeling was used to evaluate the validity of a structural/latent variable model. Several direct and indirect effects were observed among the factors under investigation. Of special importance, perceptions of nurse administration's support and perceived teaching expertise positively predicted U.S. nursing faculty members' intent to stay in the academic organization. Understanding the way that nursing faculty members' perceptions of the various factors common to the academic context interact with intent to stay in the academic organization is essential for faculty and nursing administrators. This information can assist administrators in obtaining more resources for faculty development to lobby for additional faculty in order to meet the teaching, research, and service missions of the organization; and to personalize relationships with individual faculty members to understand their needs and acknowledge their efforts.


Analytical and clinical performance of the CDC real time RT-PCR assay for detection and typing of dengue virus.
Dengue is an acute illness caused by the positive-strand RNA dengue virus (DENV). There are four genetically distinct DENVs (DENV-1-4) that cause disease in tropical and subtropical countries. Most patients are viremic when they present with symptoms; therefore, RT-PCR has been increasingly used in dengue diagnosis. The CDC DENV-1-4 RT-PCR Assay has been developed as an in-vitro diagnostic platform and was recently approved by the US Food and Drug Administration (FDA) for detection of dengue in patients with signs or symptoms of mild or severe dengue. The primers and probes of this test have been designed to detect currently circulating strains of DENV-1-4 from around the world at comparable sensitivity. In a retrospective study with 102 dengue cases confirmed by IgM anti-DENV seroconversion in the convalescent sample, the RT-PCR Assay detected DENV RNA in 98.04% of the paired acute samples. Using sequencing as a positive indicator, the RT-PCR Assay had a 97.92% positive agreement in 86 suspected dengue patients with a single acute serum sample. After extensive validations, the RT-PCR Assay performance was highly reproducible when evaluated across three independent testing sites, did not produce false positive results for etiologic agents of other febrile illnesses, and was not affected by pathological levels of potentially interfering biomolecules. These results indicate that the CDC DENV-1-4 RT-PCR Assay provides a reliable diagnostic platform capable for confirming dengue in suspected cases.


The Effects of Cinacalcet in Older and Younger Patients on Hemodialysis: The Evaluation of Cinacalcet HCl Therapy to Lower Cardiovascular Events (EVOLVE) Trial.
The calcimimetic cinacalcet reduced the risk of death or cardiovascular (CV) events in older, but not younger, patients with moderate to severe secondary hyperparathyroidism (HPT) who were receiving hemodialysis. To determine whether the lower risk in younger patients might be due to lower baseline CV risk and more frequent use of cointerventions that reduce parathyroid hormone (kidney transplantation, parathyroidectomy, and commercial cinacalcet use), this study examined the effects of cinacalcet in older (≥65 years, n=1005) and younger (<65 years, n=2878) patients. Evaluation of Cinacalcet HCl Therapy to Lower Cardiovascular Events (EVOLVE) was a global, multicenter, randomized placebo-controlled trial in 3883 prevalent patients on hemodialysis, whose outcomes included death, major CV events, and development of severe unremitting HPT. The age subgroup analysis was prespecified. Older patients had higher baseline prevalence of diabetes mellitus and CV comorbidity. Annualized rates of kidney transplantation and parathyroidectomy were >3-fold higher in younger relative to older patients and were more frequent in patients randomized to placebo. In older patients, the adjusted relative hazard (95% confidence interval) for the primary composite (CV) end point (cinacalcet versus placebo) was 0.70 (0.60 to 0.81); in younger patients, the relative hazard was 0.97 (0.86 to 1.09). Corresponding adjusted relative hazards for mortality were 0.68 (0.51 to 0.81) and 0.99 (0.86 to 1.13). Reduction in the risk of severe unremitting HPT was similar in both groups. In the EVOLVE trial, cinacalcet decreased the risk of death and of major CV events in older, but not younger, patients with moderate to severe HPT who were receiving hemodialysis. Effect modification by age may be partly explained by differences in underlying CV risk and differential application of cointerventions that reduce parathyroid hormone.


Simultaneous copy number losses within multiple subtelomeric regions in early-onset type 2 diabetes mellitus.
Genetic factors play very important roles in the onset and progression of type 2 diabetes mellitus (T2DM). However, the genetic factors correlating with T2DM onset have not as yet been fully clarified. We previously found that copy number losses in the subtelomeric region on chromosome 4p16.3 were detected in early-onset Japanese T2DM patients (onset age <35 years) at a high frequency. Herein, we additionally found two novel copy number losses within the subtelomeric regions on chromosomes 16q24.2-3 and 22q13.31-33, which have significant associations with early-onset Japanese T2DM. The associations were statistically significant by Fisher's exact tests with P values of 5.19 × 10(-3) and 1.81 × 10(-3) and odds ratios of 5.7 and 4.4 for 16q24.2-3 and 22q13.31-33, respectively. Furthermore, copy number variation (CNV) analysis of the whole genome using the CNV BeadChip system verified simultaneous copy number losses in all three subtelomeric regions in 11 of our 100 T2DM subjects, while none of 100 non-diabetic controls showed the copy number losses in all three regions. Our results suggest that the mechanism underlying induction of CNVs is involved in the pathogenesis of early-onset T2DM. Thus, copy number losses within multiple subtelomeric regions are strongly associated with early-onset T2DM and examination of simultaneous CNVs in these three regions may lead to the development of an accurate and selective procedure for detecting genetic susceptibility to T2DM.


Carbyne from first principles: chain of C atoms, a nanorod or a nanorope.
We report an extensive study of the properties of carbyne using first-principles calculations. We investigate carbyne's mechanical response to tension, bending, and torsion deformations. Under tension, carbyne is about twice as stiff as the stiffest known materials and has an unrivaled specific strength of up to 7.5 × 10(7) N·m/kg, requiring a force of ∼10 nN to break a single atomic chain. Carbyne has a fairly large room-temperature persistence length of about 14 nm. Surprisingly, the torsional stiffness of carbyne can be zero but can be "switched on" by appropriate functional groups at the ends. Further, under appropriate termination, carbyne can be switched into a magnetic semiconductor state by mechanical twisting. We reconstruct the equivalent continuum elasticity representation, providing the full set of elastic moduli for carbyne, showing its extreme mechanical performance (e.g., a nominal Young's modulus of 32.7 TPa with an effective mechanical thickness of 0.772 Å). We also find an interesting coupling between strain and band gap of carbyne, which is strongly increased under tension, from 2.6 to 4.7 eV under a 10% strain. Finally, we study the performance of carbyne as a nanoscale electrical cable and estimate its chemical stability against self-aggregation, finding an activation barrier of 0.6 eV for the carbyne-carbyne cross-linking reaction and an equilibrium cross-link density for two parallel carbyne chains of 1 cross-link per 17 C atoms (2.2 nm).


Assessment of the risk of fracture in patients with gastrointestinal disease.
Osteoporotic fractures are a common problem and associated with significant morbidity, mortality and costs. There is now increasing evidence that patients with coeliac disease are at an increased risk of osteoporotic fracture. With the advent of new therapeutic agents to reduce the risk of fracture, it is important to identify people at highest risk. The best predictors of future fracture include a previous osteoporotic fracture, low bone density, active inflammatory bowel disease, the use of oral corticosteroids and an increased risk of falling.


Human gyrovirus DNA in human blood, Italy.
Human gyrovirus (HGyV) is a recent addition to the list of agents found in humans. Prevalence, biologic properties, and clinical associations of this novel virus are still incompletely understood. We used qualitative PCRs to detect HGyV in blood samples of 301 persons from Italy. HGyV genome was detected in 3 of 100 solid organ transplant recipients and in 1 HIV-infected person. The virus was not detected in plasma samples from healthy persons. Furthermore, during observation, persons for whom longitudinal plasma samples were obtained had transient and scattered presence of circulating HGyV. Sequencing of a 138-bp fragment showed nucleotide identity among all the HGyV isolates. These results show that HGyV can be present in the blood of infected persons. Additional studies are needed to investigate possible clinical implications.


Blindness and vision impairment in a rural south Indian population: the Aravind Comprehensive Eye Survey.
To determine the prevalence of blindness and vision impairment in a rural population of southern India. A population-based cross-sectional study. A total of 17200 subjects aged 6 years or older, including 5150 subjects aged 40 years or older from 50 clusters representative of three southern districts of Tamil Nadu in southern India. All participants had preliminary screenings consisting of vision using a LogMAR illiterate E chart and anterior segment hand light examinations at the village level. Subjects aged 40 years or older were offered comprehensive eye examinations at the base hospital, including visual acuity using LogMAR illiterate E charts and refraction, slit-lamp biomicroscopy, gonioscopy, applanation tonometry, dilated fundus examinations, and automated Humphrey central 24-2 full threshold perimetry; subjects younger than 40 years of age who had any signs or symptoms of ocular disease were also offered comparable examinations at the base hospital. Visual impairment was defined as best-corrected visual acuity <6/18, and blindness was defined using both Indian (<6/60) and World Health Organization (<3/60) definitions. Comprehensive examinations at the base hospital were performed on 5150 (96.5%) of 5337 persons 40 years of age or older. Among those 40 years of age and older, presenting visual acuity at the <3/60 level was present in 4.3% (95% confidence interval [CI]: 3.8, 4.9) and 11.4% (95% CI: 10.6, 12.3) at the <6/60 level. After best correction, the corresponding figures were 1.0% (95% CI: 0.79, 1.2) and 2.1% (95% CI: 1.7, 2.5). Over 70% of subjects improved their vision by at least one line, and nearly a third by three lines after refraction. Age-related cataract was the most common potentially reversible blinding disorder (72.0%) among eyes presenting with blindness. Blindness and vision impairment remain major public health problems in India that need to be addressed. Cataracts and refractive errors remain the major reversible causes for the burden of vision impairment in this rural population.


Crack-free conditions in welding of glass by ultrashort laser pulse.
The spatial distribution of the laser energy absorbed by nonlinear absorption process in bulk glass w(z) is determined and thermal cycles due to the successive ultrashort laser pulse (USLP) is simulated using w(z) based on the transient thermal conduction model. The thermal stress produced in internal melting of bulk glass by USLP is qualitatively analyzed based on a simple thermal stress model, and crack-free conditions are studied in glass having large coefficient of thermal expansion. In heating process, cracks are prevented when the laser pulse impinges into glass with temperatures higher than the softening temperature of glass. In cooling process, shrinkage stress is suppressed to prevent cracks, because the embedded molten pool produced by nonlinear absorption process behaves like an elastic body under the compressive stress field unlike the case of CW-laser welding where the molten pool having a free surface produced by linear absorption process is plastically deformed under the compressive stress field.


Progression marker of Parkinson's disease: a 4-year multi-site imaging study.
Progression markers of Parkinson's disease are crucial for successful therapeutic development. Recently, a diffusion magnetic resonance imaging analysis technique using a bitensor model was introduced allowing the estimation of the fractional volume of free water within a voxel, which is expected to increase in neurodegenerative disorders such as Parkinson's disease. Prior work demonstrated that free water in the posterior substantia nigra was elevated in Parkinson's disease compared to controls across single- and multi-site cohorts, and increased over 1 year in Parkinson's disease but not in controls at a single site. Here, the goal was to validate free water in the posterior substantia nigra as a progression marker in Parkinson's disease, and describe the pattern of progression of free water in patients with a 4-year follow-up tested in a multicentre international longitudinal study of de novo Parkinson's disease (http://www.ppmi-info.org/). The analyses examined: (i) 1-year changes in free water in 103 de novo patients with Parkinson's disease and 49 controls; (ii) 2- and 4-year changes in free water in a subset of 46 patients with Parkinson's disease imaged at baseline, 12, 24, and 48 months; (iii) whether 1- and 2-year changes in free water predict 4-year changes in the Hoehn and Yahr scale; and (iv) the relationship between 4-year changes in free water and striatal binding ratio in a subgroup of Parkinson's disease who had undergone both diffusion and dopamine transporter imaging. Results demonstrated that: (i) free water level in the posterior substantia nigra increased over 1 year in de novo Parkinson's disease but not in controls; (ii) free water kept increasing over 4 years in Parkinson's disease; (iii) sex and baseline free water predicted 4-year changes in free water; (iv) free water increases over 1 and 2 years were related to worsening on the Hoehn and Yahr scale over 4 years; and (v) the 4-year increase in free water was associated with the 4-year decrease in striatal binding ratio in the putamen. Importantly, all longitudinal results were consistent across sites. In summary, this study demonstrates an increase over 1 year in free water in the posterior substantia nigra in a large cohort of de novo patients with Parkinson's disease from a multi-site cohort study and no change in healthy controls, and further demonstrates an increase of free water in Parkinson's disease over the course of 4 years. A key finding was that results are consistent across sites and the 1-year and 2-year increase in free water in the posterior substantia nigra predicts subsequent long-term progression on the Hoehn and Yahr staging system. Collectively, these findings demonstrate that free water in the posterior substantia nigra is a valid, progression imaging marker of Parkinson's disease, which may be used in clinical trials of disease-modifying therapies.


Host Immune-Metabolic Adaptations Upon Mycobacterial Infections and Associated Co-Morbidities.
Mycobacterial diseases are a major public health challenge. Their causative agents include, in order of impact, members of the Mycobacterium tuberculosis complex (causing tuberculosis), Mycobacterium leprae (causing leprosy), and non-tuberculous mycobacterial pathogens including Mycobacterium ulcerans. Macrophages are mycobacterial targets and they play an essential role in the host immune response to mycobacteria. This review aims to provide a comprehensive understanding of the immune-metabolic adaptations of the macrophage to mycobacterial infections. This metabolic rewiring involves changes in glycolysis and oxidative metabolism, as well as in the use of fatty acids and that of metals such as iron, zinc and copper. The macrophage metabolic adaptations result in changes in intracellular metabolites, which can post-translationally modify proteins including histones, with potential for shaping the epigenetic landscape. This review will also cover how critical tuberculosis co-morbidities such as smoking, diabetes and HIV infection shape host metabolic responses and impact disease outcome. Finally, we will explore how the immune-metabolic knowledge gained in the last decades can be harnessed towards the design of novel diagnostic and therapeutic tools, as well as vaccines.


In vitro fertilization (IVF) in mammals: epigenetic and developmental alterations. Scientific and bioethical implications for IVF in humans.
The advent of in vitro fertilization (IVF) in animals and humans implies an extraordinary change in the environment where the beginning of a new organism takes place. In mammals fertilization occurs in the maternal oviduct, where there are unique conditions for guaranteeing the encounter of the gametes and the first stages of development of the embryo and thus its future. During this period a major epigenetic reprogramming takes place that is crucial for the normal fate of the embryo. This epigenetic reprogramming is very vulnerable to changes in environmental conditions such as the ones implied in IVF, including in vitro culture, nutrition, light, temperature, oxygen tension, embryo-maternal signaling, and the general absence of protection against foreign elements that could affect the stability of this process. The objective of this review is to update the impact of the various conditions inherent in the use of IVF on the epigenetic profile and outcomes of mammalian embryos, including superovulation, IVF technique, embryo culture and manipulation and absence of embryo-maternal signaling. It also covers the possible transgenerational inheritance of the epigenetic alterations associated with assisted reproductive technologies (ART), including its phenotypic consequences as is in the case of the large offspring syndrome (LOS). Finally, the important scientific and bioethical implications of the results found in animals are discussed in terms of the ART in humans.


The impact of daily setup variations on head-and-neck intensity-modulated radiation therapy.
Intensity-modulated radiation therapy (IMRT) in the treatment of head-and-neck (H&N) cancer provides the opportunity to diminish normal tissue toxicity profiles and thereby enhance patient quality of life. However, highly conformal treatment techniques commonly establish steep dose gradients between tumor and avoidance structures. Daily setup variations can therefore significantly compromise the ultimate precision of idealized H&N IMRT delivery. This study provides a detailed analysis regarding the potential impact of daily setup variations on the overall integrity of H&N IMRT. A series of 10 patients with advanced H&N cancer were prospectively enrolled in a clinical trial to examine daily H&N radiation setup accuracy. These patients were treated with conventional shrinking field design using three-dimensional treatment planning techniques (not IMRT). Immobilization and alignment were performed using modern H&N practice techniques including conventional thermoplastic masking, baseplate fixation to the treatment couch, three-point laser alignment, and weekly portal film evaluation. After traditional laser alignment, setup accuracy was assessed daily for each patient by measuring 3 Cartesian and 3 angular deviations from the specified isocenter using a high-precision, optically guided patient localization system, which affords submillimeter setup accuracy. These positional errors were then applied to a distinct series of 10 H&N IMRT plans for detailed analysis regarding the impact of daily setup variation (without optical guidance) on the ultimate integrity of IMRT plans over a 30-day treatment course. Dose-volume histogram (DVH), equivalent uniform dose (EUD), mean total dose (mTd), and maximal total dose (MTD) for normal structures were analyzed for IMRT plans with and without incorporation of daily setup variation. Using conventional H&N masking and laser alignment for daily positioning, the mean setup error in any single dimension averaged 3.33 mm. However, when all six degrees of freedom were accounted for, using the optically guided patient localization system, the mean composite vector offset was 6.97 mm with a standard deviation of 3.63 mm. Superimposition of mean offset vectors on idealized H&N IMRT treatment plans enabled evaluation of resultant shifts in DVH, EUD, mTd, and MTD calculations. Partial geographic tumor miss (GTV underdosing) and normal tissue overdosing was common when these mean positional offsets were incorporated. The decrease in EUD for defined tumor volumes ranged up to 21% when the largest offset histories were applied, and 3-14% for plans when the least and median offset histories were applied. The successful implementation of H&N IMRT requires accurate and reproducible treatment delivery over a 6- to 7-week treatment course. The adverse impact of daily setup variation, which occurs routinely with conventional H&N masking techniques, may be considerably greater than recognized. Isocenter verification checks on two-dimensional orthogonal films may not sufficiently alert the clinician to the magnitude of three-dimensional offset vectors and the resultant impact on the quality of overall IMRT delivery. Unrecognized geographic miss and resultant target underdose may occur. Similarly, selected normal structures such as parotid glands may receive higher doses than intended. The results of this study suggest that more rigorous immobilization techniques than conventional masking and routine patient setup tracking methodologies are important for the accurate monitoring and successful delivery of high-quality IMRT for H&N cancer.


All disease begins in the (leaky) gut: role of zonulin-mediated gut permeability in the pathogenesis of some chronic inflammatory diseases.
Improved hygiene leading to reduced exposure to microorganisms has been implicated as one possible cause for the recent "epidemic" of chronic inflammatory diseases (CIDs) in industrialized countries. That is the essence of the hygiene hypothesis that argues that rising incidence of CIDs may be, at least in part, the result of lifestyle and environmental changes that have made us too "clean" for our own good, so causing changes in our microbiota. Apart from genetic makeup and exposure to environmental triggers, inappropriate increase in intestinal permeability (which may be influenced by the composition of the gut microbiota), a "hyper-belligerent" immune system responsible for the tolerance-immune response balance, and the composition of gut microbiome and its epigenetic influence on the host genomic expression have been identified as three additional elements in causing CIDs. During the past decade, a growing number of publications have focused on human genetics, the gut microbiome, and proteomics, suggesting that loss of mucosal barrier function, particularly in the gastrointestinal tract, may substantially affect antigen trafficking, ultimately influencing the close bidirectional interaction between gut microbiome and our immune system. This cross-talk is highly influential in shaping the host gut immune system function and ultimately shifting genetic predisposition to clinical outcome. This observation led to a re-visitation of the possible causes of CIDs epidemics, suggesting a key pathogenic role of gut permeability. Pre-clinical and clinical studies have shown that the zonulin family, a group of proteins modulating gut permeability, is implicated in a variety of CIDs, including autoimmune, infective, metabolic, and tumoral diseases. These data offer novel therapeutic targets for a variety of CIDs in which the zonulin pathway is implicated in their pathogenesis.


Symptom severity of bipolar disorder during the menopausal transition.
Little is known about the mood symptom experience of women with bipolar disorder during the menopausal transition (MT). Yet times of rapid hormonal decline, such as the postpartum, are associated with increased risk of severe mood episodes in bipolar disorder, and the MT is a time of increased risk for unipolar depression in women with or without a history of depression. Enrollment included 56 women 40-60 years old diagnosed in the bipolar spectrum who were experiencing menopausal symptoms or were up to 5 years since their final menstrual period. Menopausal stages included early menopause, late menopause, or early postmenopause based on standardized criteria. Observational, prospective standardized mood symptom and reproductive hormone assessments were completed periodically. Concurrent menopausal symptoms as well as history of mood exacerbation during past reproductive events were assessed. Forty-four women were included in the main analysis. The average Montgomery-Asberg Depression Rating Scale (MADRS) score was 4.43 points higher in the late transition/early postmenopausal stage women (n = 29) compared to the early menopausal stage women (n = 15) (±SE 2.14; p = 0.039), corresponding to a roughly 10 % higher score (range 0-40) in the late/post stage across all study visits. Results were similar for the Young Mania Rating Scale (YMRS), where the average score was 2.54 points higher in the late/early postmenopausal stage women compared to the early menopausal stage women (±SE 1.15; p = 0.027), also roughly 10 % higher (range 0-26). Estradiol and follicle-stimulating hormone (FSH) absolute levels as well as between-visit change in levels were not notably associated with YMRS or MADRS during study observation. Total Greene Climacteric Symptom (menopausal symptom) score was significantly associated with MADRS but not YMRS. History of mood exacerbation premenstrually and/or postpartum was not significantly associated with YMRS or MADRS severity during the MT. These results support the theory that times of increased reproductive hormonal changes, such as the late MT and early postmenopause, here compared to early MT, are associated with greater mood symptom severity in bipolar spectrum women. Nonetheless, absolute or change in FSH and estradiol levels were not significantly associated with depression or mood elevation severity.


TLC1: template RNA component of Saccharomyces cerevisiae telomerase.
Telomeres, the natural ends of linear eukaryotic chromosomes, are essential for chromosome stability. Because of the nature of DNA replication, telomeres require a specialized mechanism to ensure their complete duplication. Telomeres are also capable of silencing the transcription of genes that are located near them. In order to identify genes in the budding yeast Saccharomyces cerevisiae that are important for telomere function, a screen was conducted for genes that, when expressed in high amounts, would suppress telomeric silencing. This screen lead to the identification of the gene TLC1 (telomerase component 1). TLC1 encodes the template RNA of telomerase, a ribonucleoprotein required for telomere replication in a variety of organisms. The discovery of TLC1 confirms the existence of telomerase in S. cerevisiae and may facilitate both the analysis of this enzyme and an understanding of telomere structure and function.


Delivering trauma training to multiple health-worker cadres in nine sub-Saharan African countries: lessons learnt from the COOL programme.
Africa has one of the highest road-traffic mortality rates in the world. Nurses and clinical officers play a pivotal part in trauma care as a result of substantial shortage of doctors. The COOL (COSECSA-Oxford-Orthopaedic-Link) programme has delivered primary trauma care (PTC) training in nine sub-Saharan African countries across a wide cadre of health-workers (540 doctors, 260 nurses, 119 clinical officers, and 111 medical students). This prospective study investigates the effect of 28 consecutive PTCs and the training challenges that exist between different cadres and health institutions. The course trains delegates in key trauma concepts: primary survey, airway management, chest injuries, major haemorrhage, and paediatric trauma. Candidates' knowledge of these concepts was assessed before and after the course with a validated 30 Single-Best-Answer multiple choice questionnaire. Assessment scores were analysed by cadre, urban (383 candidates) or rural institutions (647 candidates), and sex (657 men, 373 women). A concept was categorised as being poorly understood when half the candidates achieved less than 50% of the correct answers. Descriptive statistics and MANOVA analysis were used, with an alpha level set at 0·05. 1030 PTC providers were trained between Dec 5, 2012, and Dec 19, 2013. There was significant increase in multiple choice questionnaire (58% to 77%, p<0·05) and clinical confidence (68% to 90%, p<0·05) scores among delegates post course, with independent covariants of institution location and cadre significantly affecting post-course scores. Doctors achieved satisfactory scores on all key concepts (67% to 84%, p<0·05). Clinical officers (all concepts 53% to 76%, p<0·05) particularly struggled with paediatric trauma (94 candidates <50%, mean 24·23 [95% CI 19-30]). Nurses (all concepts 42% to 64%, p<0·05) had difficulty with chest injuries (203 pre-course to 153 post-course candidates <50%, mean 49% [95% CI 45-52]) and paediatric trauma (212 pre-course to 161 post-course candidates ≤50%, post course mean 46% [95% CI 43-53]). Medical students achieved satisfactory scores in all concepts (overall 53% to 74%, p<0·05). Health-workers based in urban hospitals (82%) outperformed those in rural hospitals (72%) (p=0·001) and sex had no significant effect on performance (p=0·07). Our study shows that PTC courses led to improvement in trauma management knowledge and clinical confidence among a wide cadre of health-workers. However, these are new concepts for many front-line health-workers, and regular refresher training will be required. There is also a difference in understanding of key trauma concepts among the different cadres. Future training in this region should address areas of weakness unique to each cadre, particularly paediatric trauma care. Health Partnership Scheme through the UK Department for International Development (DFID).


Rate-dependent effects of monoamine releasers on intracranial self-stimulation in rats: implications for abuse liability assessment.
'Rate dependency' in the discipline of behavioral pharmacology describes a phenomenon wherein the effect of a drug on the rate of a behavior varies systematically as a function of the baseline, predrug rate of that behavior. Historically, rate-dependency studies have compared drug effects on different baseline rates of behavior maintained either by different schedules of reinforcement or during sequential segments of a fixed-interval schedule. The current experiment generated different baseline rates of behavior by altering frequency of electrical stimulation in an intracranial self-stimulation assay. Amphetamine and 10 other monoamine releasers were analyzed for their ability to produce rate-dependent effects in this assay. There were three main findings. First, all compounds produced rate-dependent effects at some dose. Second, one parameter of rate-dependency plots (peak Y-intercept of the regression line) correlated with in-vitro neurochemical data on selectivity of these compounds to release dopamine versus serotonin (P<0.025, R=0.50). Lastly, a correlation between peak Y-intercept and breakpoints under a progressive-ratio procedure in nonhuman primates was also significant (P<0.05, R=0.64). Overall, these results extend the rate-dependent effects of monoamine releasers to behavior maintained under intracranial self-stimulation and suggest that, at least for monoamine releasers, the Y-intercept parameter of rate-dependency plots might be a useful metric of drug reward and predictor of drug self-administration measures of drug reinforcement.


Electromagnetic waves in a topological insulator thin film stack: helicon-like wave mode and photonic band structure.
We theoretically explore the electromagnetic modes specific to a topological insulator superlattice in which topological and conventional insulator thin films are stacked periodically. In particular, we obtain analytic formulas for low energy mode that corresponds to a helicon wave, as well as those for photonic bands. We illustrate that the system can be modeled as a stack of quantum Hall layers whose conductivity tensors alternately change signs, and then we analyze the photonic band structures. This subject is a natural extension of a previous study by Tselis et al., which took into consideration a stack of identical quantum Hall layers but their discussion was limited into a low energy mode. Thus we provide analytic formulas for photonic bands and compare their features between the two systems. Our central findings in the topological insulator superlattice are that a low energy mode corresponding to a helicon wave has linear dispersion instead of the conventional quadratic form, and that a robust gapless photonic band appears although the system considered has spacial periodicity. In addition, we demonstrate that the photonic bands agree with the numerically calculated transmission spectra.


Long-term outcome of hematuria home screening for bladder cancer in men.
The objectives of this study were to determine whether bladder cancer (BC) screening in healthy men could lead to earlier detection and reduced BC mortality compared with unscreened men and to determine long-term outcomes of a geographically defined, unscreened population with newly diagnosed BC. In 1987 and from 1998 to 1992, 1575 men ages 50 years and older who were solicited from well patient rosters in clinics in and around Madison, Wisconsin, tested their urine repetitively with a chemical reagent strip for hemoglobin. Participants who had positive test results underwent standard urologic evaluation. BC grades and stages and the outcomes of men with BC detected by screening were compared with the grades, stages, and outcomes of 87% of men ages 50 years and older with newly diagnosed BC who were reported to the Wisconsin Tumor Registry in 1988 (n = 509 men). Two hundred fifty-eight screening participants (16.4%) were evaluated for hematuria, and 21 participants (8.1%) were diagnosed with BC. Proportions of low-grade (Grade 1 and 2) superficial (Stage Ta and T1) versus high-grade (Grade 3) superficial or invasive (Stage > or = T2) cancers in screened men (52.4% vs. 47.7%) and in men from the tumor registry (60.3% vs. 39.7%) were similar (P = .50). The proportion of high-grade superficial or invasive BCs that were invasive were lower in screened men (10%) than in unscreened men (60%; P = .002). At 14 years of follow-up, no men with screen-detected BC had died of BC, whereas 20.4% of men with unscreened BC had died of BC (P = .02). Screening effected the early detection of BC and may reduce mortality from BC compared with BC that is diagnosed at standard clinical presentation.


Statistical algorithms improve accuracy of gene fusion detection.
Gene fusions are known to play critical roles in tumor pathogenesis. Yet, sensitive and specific algorithms to detect gene fusions in cancer do not currently exist. In this paper, we present a new statistical algorithm, MACHETE (Mismatched Alignment CHimEra Tracking Engine), which achieves highly sensitive and specific detection of gene fusions from RNA-Seq data, including the highest Positive Predictive Value (PPV) compared to the current state-of-the-art, as assessed in simulated data. We show that the best performing published algorithms either find large numbers of fusions in negative control data or suffer from low sensitivity detecting known driving fusions in gold standard settings, such as EWSR1-FLI1. As proof of principle that MACHETE discovers novel gene fusions with high accuracy in vivo, we mined public data to discover and subsequently PCR validate novel gene fusions missed by other algorithms in the ovarian cancer cell line OVCAR3. These results highlight the gains in accuracy achieved by introducing statistical models into fusion detection, and pave the way for unbiased discovery of potentially driving and druggable gene fusions in primary tumors.


Transcranial direct current stimulation (tDCS) combined with cognitive training in adolescent boys with ADHD: a double-blind, randomised, sham-controlled trial.
Transcranial direct current stimulation (tDCS) could be a side-effect-free alternative to psychostimulants in attention-deficit/hyperactivity disorder (ADHD). Although there is limited evidence for clinical and cognitive effects, most studies were small, single-session and stimulated left dorsolateral prefrontal cortex (dlPFC). No sham-controlled study has stimulated the right inferior frontal cortex (rIFC), which is the most consistently under-functioning region in ADHD, with multiple anodal-tDCS sessions combined with cognitive training (CT) to enhance effects. Thus, we investigated the clinical and cognitive effects of multi-session anodal-tDCS over rIFC combined with CT in double-blind, randomised, sham-controlled trial (RCT, ISRCTN48265228). Fifty boys with ADHD (10-18 years) received 15 weekday sessions of anodal- or sham-tDCS over rIFC combined with CT (20 min, 1 mA). ANCOVA, adjusting for baseline measures, age and medication status, tested group differences in clinical and ADHD-relevant executive functions at posttreatment and after 6 months. ADHD-Rating Scale, Conners ADHD Index and adverse effects were significantly lower at post-treatment after sham relative to anodal tDCS. No other effects were significant. This rigorous and largest RCT of tDCS in adolescent boys with ADHD found no evidence of improved ADHD symptoms or cognitive performance following multi-session anodal tDCS over rIFC combined with CT. These findings extend limited meta-analytic evidence of cognitive and clinical effects in ADHD after 1-5 tDCS sessions over mainly left dlPFC. Given that tDCS is commercially and clinically available, the findings are important as they suggest that rIFC stimulation may not be indicated as a neurotherapy for cognitive or clinical remediation for ADHD.


Similarities between protein IIIa and protein IIIb, two prominent synaptic vesicle-associated phosphoproteins.
Protein IIIa (Mr 74,000) and protein IIIb (Mr 55,000) are 2 major phosphoproteins found in mammalian brain. It was previously shown in intact nerve cells that the phosphorylation state of these 2 proteins could be increased by electrical stimulation, by depolarizing agents in the presence of calcium, and by 8-bromo-cAMP. We now report that protein IIIa and protein IIIb possess significant structural homology, as indicated by immunochemical studies using polyclonal and monoclonal antibodies and by peptide-mapping studies. A quantitative radioimmunoassay using immunolabeling in SDS-polyacrylamide gels has been used to study the tissue distribution and regional and subcellular distribution in the brain of the 2 proteins. The 2 proteins were found only in nervous tissue and the adrenal medulla. Subcellular fractionation studies suggested that both proteins are associated with synaptic vesicles.


Inhibition of lipopolysaccharide transport to the outer membrane in Pseudomonas aeruginosa by peptidomimetic antibiotics.
The asymmetric outer membrane (OM) of Gram-negative bacteria contains lipopolysaccharide (LPS) in the outer leaflet and phospholipid in the inner leaflet. During OM biogenesis, LPS is transported from the periplasm into the outer leaflet by a complex comprising the OM proteins LptD and LptE. Recently, a new family of macrocyclic peptidomimetic antibiotics that interact with LptD of the opportunistic human pathogen Pseudomonas aeruginosa was discovered. Here we provide evidence that the peptidomimetics inhibit the LPS transport function of LptD. One approach to monitor LPS transport involved studies of lipid A modifications. Some modifications occur only in the inner membrane while others occur only in the OM, and thus provide markers for LPS transport within the bacterial envelope. We prepared a conditional lptD mutant of P. aeruginosa PAO1 that allowed control of lptD expression from the rhamnose promoter. With this mutant, the effects caused by the antibiotic on the wild-type strain were compared with those caused by depleting LptD in the mutant strain. When LptD was depleted in the mutant, electron microscopy revealed accumulation of membrane-like material within cells and OM blebbing; this mirrored similar effects in the wild-type strain caused by the antibiotic. Moreover, the bacterium responded to the antibiotic, and to depletion of LptD, by introducing the same lipid A modifications, consistent with inhibition by the antibiotic of LptD-mediated LPS transport. This conclusion was further supported by monitoring the radiolabelling of LPS from [¹⁴C]acetate, and by fractionation of IM and OM components. Overall, the results provide support for a mechanism of action for the peptidomimetic antibiotics that involves inhibition of LPS transport to the cell surface.