Purine oversecretion in cultured murine lymphoma cells deficient in adenylosuccinate synthetase: genetic model for inherited hyperuricemia and gout.
Alterations in several specific enzymes have been associated with increased rates of purine synthesis de novo in human and other mammalian cells. However, these recognized abnormalities in humans account for only a few percent of the clinical cases of hyperuricemia and gout. We have examined in detail the rates of purine production de novo and purine excretion by normal and by mutant (AU-100) murine lymphoma T cells (S49) 80% deficient in adenylosuccinate synthetase [IMP:L-aspartate ligase (GDP-forming), EC 184.108.40.206]. The intracellular ATP concentration of the mutant cells is slightly diminished, but their GTP is increased 50% and their IMP, four-fold. Compared to wild-type cells, the AU-100 cells excrete into the culture medium 30- to 50-fold greater amounts of purine metabolites consisting mainly of inosine. Moreover, the AU-100 cell line overproduces total purines. In an AU-100-derived cell line, AU-TG50B, deficient in adenylosuccinate synthetase and hypoxanthine/guanine phosphoribosyltransferase (IMP:pyrophosphate phosphoribosyltransferase, EC 184.108.40.206), purine nucleoside excretion is increased 50- to 100-fold, and de novo synthesis is even greater than that for AU-100 cells. The overexcretion of purine metabolites by the AU-100 cells seems to be due to the primary genetic deficiency of adenylosuccinate synthetase, a deficiency that requires the cell to increase intracellular IMP in an attempt to maintain ATP levels. As a consequence of elevated IMP pools, large amounts of inosine are secreted into the culture medium. We propose that a similar primary genetic defect may account for the excessive purine excretion in some patients with dominantly inherited hyperuricemia and gout.


Developmental meta-analysis of the functional neural correlates of autism spectrum disorders.
There is a pressing need to elucidate the brain-behavior interactions underlying autism spectrum disorders (ASD) given the marked rise in ASD diagnosis over the past decade. Functional magnetic resonance imaging (fMRI) has begun to address this need, but few fMRI studies have evaluated age-related changes in ASD. Therefore, we conducted a developmental analysis of activation likelihood estimation (ALE) meta-analysis to compare child versus adult ASD fMRI studies. We hypothesized that children and adolescents with ASD (<18 years old) would rely less on prefrontal cortex structures than adults (≥18 years old). PubMed and PsycInfo literature searches were conducted to identify task-dependent fMRI studies of children or adults with ASD. Then recent GingerALE software improvements were leveraged to perform direct comparisons of child (n = 18) versus adult (n = 24) studies. ALE meta-analyses of social tasks showed that children and adolescents with ASD versus adults had significantly greater hyperactivation in the left post-central gyrus, and greater hypoactivation in the right hippocampus and right superior temporal gyrus. ALE meta-analyses of nonsocial tasks showed that children with ASD versus adults had significantly greater hyperactivation in the right insula and left cingulate gyrus, and hypoactivation in the right middle frontal gyrus. Our data suggest that the neural alterations associated with ASD are not static, occurring only in early childhood. Instead, children with ASD have altered neural activity compared to adults during both social and nonsocial tasks, especially in fronto-temporal structures. Longitudinal neuroimaging studies are required to examine these changes prospectively, as potential targets for brain-based treatments for ASD.


Insights into phases of liquid water from study of its unusual glass-forming properties.
The vitrification of pure water is compared with that of molecular solutions rich in water, and gross differences are noted. Thermodynamic reasoning and direct observations on noncrystallizing nanoconfined water indicate that the glass transition in ambient-pressure water is qualitatively distinct from that found in the usual molecular liquids. It belongs instead to the order-disorder class of transition seen in molecular and ionic crystalline materials. The distinctive "folding funnel" energy landscape for this type of system explains the extreme weakness of the glass transition of water as well as the consequent confusion that has characterized its scientific history; it also explains the very small excess entropy at the glass transition temperature. The relation of confined water behavior to that of bulk is discussed, and the "fragile-to-strong" transition for supercooled water is interpreted by adding a "critical point-free" scenario to the two competing scenarios for understanding supercooled bulk water.


Randomized controlled trial of a breath-activated nebulizer in patients with exacerbation of COPD.
Exacerbations of COPD (ECOPD) are characterized by increased dyspnea due to dynamic pulmonary hyperinflation. This study sought to determine whether the AeroEclipse II breath-activated nebulizer (BAN) would produce greater bronchodilator responses than a continuous flow small-volume nebulizer (SVN) in patients with ECOPD. Prospective randomized controlled trial. Forty patients with ECOPD were recruited to participate in the trial. The primary study outcomes were inspiratory capacity (IC) and dyspnea via the Borg scale. Subjects were randomized to receive bronchodilator from either a BAN or a continuous flow SVN. Subjects in both groups received 2.5 mg albuterol sulfate and 0.5 mg ipratropium bromide by nebulizer every 4 hours, and 2.5 mg albuterol every 2 hours as needed. Approximately 2 hours after the subject's 6th scheduled nebulizer treatment, IC, dyspnea, and respiratory frequency measurements were repeated. Both groups received an equal number of nebulizer treatments over the study period (BAN 6.25 ± 0.55, control 6.2 ± 0.7, P = .80). Following completion of the study protocol the BAN group had a higher IC than the SVN group (1.83 ± 0.65 L vs 1.42 ± 0.49 L, P = .03, respectively). The change in IC was higher in the BAN group (0.33 ± 0.31 L than in the SVN group (0.15 ± 0.19 L, P = .03). The BAN group also had a lower respiratory rate (19 ± 3.3 breaths/min vs 22 ± 5.3 breaths/min, P = .03, respectively). There was no difference in resting dyspnea as measured with the Borg scale (BAN 3.3 ± 2.1, SVN 3.5 ± 2.4, P = .69) or stay (BAN 4.6 ± 2.6 d, SVN 5.7 ± 2.8 d, P = .21). In this cohort of patients with ECOPD, a BAN was more effective in reducing lung hyperinflation and respiratory frequency than a continuous-flow SVN.


Trends in peptic ulcer mortality in Italy, 1955-1985.
The aim was to analyse trends in mortality from peptic ulcer in Italy between 1955 and 1985, disentangling the role of age, cohort of birth, and period of death. This was a descriptive epidemiological survey. Death certification numbers from peptic ulcer and estimates of the resident population were obtained from official sources. From these data, age specific and age standardised mortality rates from peptic ulcer were computed. A log-linear age, period, and cohort model with arbitrary constraints on the parameters was applied to the matrices of age specific rates between 25 and 74 years of age. This was a national survey. For males, overall peptic ulcer mortality was constant or moderately upwards (from 9.0 to 9.4/100,000, on the basis of World Standard Population) from 1955 to the mid-1970s, but declined considerably afterwards to 3.7/100,000 in 1985. Truncated (35-64 years) rates were slightly more favourable, even in earlier calendar periods, but a substantial drop was observed only from the late 1970s onwards. In 1985 the standardised rate was 3.3/100,000 compared to 18.3 in 1955-1959. For females, the time pattern was similar, although the extent of the decline was smaller in absolute terms (from 1.6 to 1.2/100,000, all ages; from 2.3 to 0.7, truncated). Age specific rates showed an earlier decline in the young, while the fall started only after the mid-1970s in older age groups. On the basis of a log-linear age, period, and cohort model, the Italian generations born in the first decade of this century had the highest risk of dying from peptic ulcer, with a substantial decline for each subsequent cohort. Period trends were stable between 1955 and the mid-1970s, but declined appreciably afterwards. In Italy the peak rate of peptic ulcer mortality was observed in the early 1970s, with a delay of around two decades in comparison with northern Europe and the USA. This can be related to the later process of industrialization in Italy, with the consequent changes in lifestyle habits, and to a later pattern of rise and decline of cigarette smoking. A likely explanation of the falls in mortality on a period of death basis over the last decade is the introduction of new drugs (histamine-2 receptor antagonists) for the treatment of peptic ulcer, with a reduction of complications of the disease and related mortality. This decline in mortality from peptic ulcer corresponds to the avoidance of over 1500 deaths per year in the whole of Italy.


Natural exposure to ultraviolet-B radiation in two species of chameleons from Madagascar.
Many reptiles require ultraviolet-B radiation between 290 and 315 nm (UV-B) to synthesize vitamin D3 and process dietary calcium. In captivity, exposure to too little or too much UV-B can result in health problems such as metabolic bone disease. While it is recognized that UV-B is necessary to successfully maintain many reptiles in captivity, the actual levels of UV-B that species are exposed to in nature is poorly known. We measured the UV-B exposure of two species of chameleon (Calumma brevicorne and C. nasutum) in the field in Madagascar over a period of four months. We found that both species were exposed to less UV radiation than that which was available in full sun. Only on rare occasions were chameleons observed in areas with a UV Index (UVI) greater than 3.0, and the median UVI for both species was only 0.3. There was no daily temporal pattern in UV exposure for C. nasutum, but C. brevicorne was found in areas with lower UV levels in the late afternoon when compared to late morning. Additionally, C. nasutum males showed higher UV exposure than females in late morning. Our results suggest that both C. brevicorne and C. nasutum can be classified as Ferguson Zone 1 species, and should be provided with a UV-B gradient in captivity that offers access to UV-B radiation as well as adequate shaded refuge.


SREBF1 Activity Is Regulated by an AR/mTOR Nuclear Axis in Prostate Cancer.
Reprogramming of cellular metabolism is an important feature of prostate cancer, including altered lipid metabolism. Recently, it was observed that the nuclear fraction of mTOR is essential for the androgen-mediated metabolic reprogramming of prostate cancer cells. Herein, it is demonstrated that the androgen receptor (AR) and mTOR bind to regulatory regions of sterol regulatory element-binding transcription factor 1 (SREBF1) to control its expression, whereas dual activation of these signaling pathways also promotes SREBF1 cleavage and its translocation to the nucleus. Consequently, SREBF1 recruitment to regulatory regions of its target genes is induced upon treatment with the synthetic androgen R1881, an effect abrogated upon inhibition of the mTOR signaling pathway. In turn, pharmacologic and genetic inhibition of SREBF1 activity impairs the androgen-mediated induction of the key lipogenic genes fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD1). Consistent with these observations, the expression of the SREBF1, FASN, and SCD1 genes is significantly correlated in human prostate cancer tumor clinical specimens. Functionally, blockade of SREBF1 activity reduces the androgen-driven lipid accumulation. Interestingly, decreased triglyceride accumulation observed upon SREBF1 inhibition is paralleled by an increase in mitochondrial respiration, indicating a potential rewiring of citrate metabolism in prostate cancer cells. Altogether, these data define an AR/mTOR nuclear axis, in the context of prostate cancer, as a novel pathway regulating SREBF1 activity and citrate metabolism.Implications: The finding that an AR/mTOR complex promotes SREBF1 expression and activity enhances our understanding of the metabolic adaptation necessary for prostate cancer cell growth and suggests novel therapeutic approaches to target metabolic vulnerabilities in tumors. Mol Cancer Res; 16(9); 1396-405. ©2018 AACR.


Comparative riverscape genomics of the rainbow darter (Etheostoma caeruleum) in glaciated and unglaciated environments.
Periodic glaciation during the Quaternary period shaped the contemporary riverscape and distribution of freshwater fishes in the Mississippi River drainage of central North America. The rainbow darter (Etheostoma caeruleum) is a member of this ichthyofauna and has a disjunct distribution in glaciated and unglaciated environments west of the Mississippi River. Based on glacial history of the region, there are different expectations on the observed spatial genetic structure of populations in these environments. The aim of this study was to utilize genome-wide SNP data to compare the population genomic structure of the rainbow darter in river networks with disparate glacial histories; the Volga River in the glaciated upper Mississippi River basin and the Meramec River in the unglaciated Ozark Plateau. Individuals were sampled from localities within each river system at distances dictated by the organismal life history and habitat preferences. Riverscape analyses were performed on three datasets: total combined localities of both rivers and one for each river independently. The results revealed a lasting influence of historic glaciation on the population genomic structure of rainbow darter populations. There was evidence of population expansion into the glaciated northern region following glacial retreat. The population genetic signature within the Volga River did not fit expectations of the stream hierarchy model, but revealed a pattern of repeated colonization and extirpation due to cyclic glaciation. The population within the unglaciated Meramec River adhered to the stream hierarchy model, with a directional order of genetic diversity based on the life history and habitat preferences of the species. These results demonstrate the importance of considering the geologic and climatic history of a region as well as the life history of an organism when interpreting spatial genetic patterns.


Systematic and intestinal antibody-secreting cell responses and correlates of protective immunity to human rotavirus in a gnotobiotic pig model of disease.
Neonatal gnotobiotic pigs orally inoculated with virulent (intestinal-suspension) Wa strain human rotavirus (which mimics human natural infection) developed diarrhea, and most pigs which recovered (87% protection rate) were immune to disease upon homologous virulent virus challenge at postinoculation day (PID) 21. Pigs inoculated with cell culture-attenuated Wa rotavirus (which mimics live oral vaccines) developed subclinical infections and seroconverted but were only partially protected against challenge (33% protection rate). Isotype-specific antibody-secreting cells (ASC were enumerated at selected PID in intestinal (duodenal and ileal lamina propria and mesenteric lymph node [MLN]) and systemic (spleen and blood) lymphoid tissues by using enzyme-linked immunospot assays. At challenge (PID 21), the numbers of virus-specific immunoglobulin A (IgA) ASC, but not IgG ASC, in intestines and blood were significantly greater in virulent-Wa rotavirus-inoculated pigs than in attenuated-Wa rotavirus-inoculated pigs and were correlated (correlation coefficients: for duodenum and ileum, 0.9; for MLN, 0.8; for blood, 0.6) with the degree of protection induced. After challenge, the numbers of IgA and IgG virus-specific ASC and serum-neutralizing antibodies increased significantly in the attenuated-Wa rotavirus-inoculated pigs but not in the virulent-Wa rotavirus-inoculated pigs (except in the spleen and except for IgA ASC in the duodenum). The transient appearance of IgA ASC in the blood mirrored the IgA ASC responses in the gut, albeit at a lower level, suggesting that IgA ASC in the blood of humans could serve as an indicator for IgA ASC responses in the intestine after rotavirus infection. To our knowledge, this is the first report to study and identify intestinal IgA ASC as a correlate of protective active immunity in an animal model of human-rotavirus-induced disease.


Strong heterogeneity in nucleotidic composition and codon bias in the pea aphid (Acyrthosiphon pisum) shown by EST-based coding genome reconstruction.
The aim of this study was to analyze patterns of nucleotidic composition and codon usage in the pea aphid genome (Acyrthosiphon pisum). A collection of 60,000 expressed sequence tags (ESTs) in the pea aphid has been used to automatically reconstruct 5809 coding sequences (CDSs), based on similarity with known proteins and on coding style recognition. Reconstructions were manually checked for ribosomal proteins, leading to tentatively reconstruct the nea-complete set of this category. Pea aphid coding sequences showed a shift toward AT (especially at the third codon position) compared to drosophila homologues. Genes with a putative high level of expression (ribosomal and other genes with high EST support) remained more GC3-rich and had a distinct codon usage from bulk sequences: they exhibited a preference for C-ending codons and CGT (for arginine), which thus appeared optimal for translation. However, the discrimination was not as strong as in drosophila, suggesting a reduced degree of translational selection. The space of variation in codon usage for A. pisum appeared to be larger than in drosophila, with a substantial fraction of genes that remained GC3-rich. Some of those (in particular some structural proteins) also showed high levels of codon bias and a very strong preference for C-ending codons, which could be explained either by strong translational selection or by other mechanisms. Finally, genomic traces were analyzed to build 206 fragments containing a full CDS, which allowed studying the correlations between GC contents of coding and those of noncoding (flanking and introns) sequences.


Human islets contain four distinct subtypes of β cells.
Human pancreatic islets of Langerhans contain five distinct endocrine cell types, each producing a characteristic hormone. The dysfunction or loss of the insulin-producing β cells causes diabetes mellitus, a disease that harms millions. Until now, β cells were generally regarded as a single, homogenous cell population. Here we identify four antigenically distinct subtypes of human β cells, which we refer to as β1-4, and which are distinguished by differential expression of ST8SIA1 and CD9. These subpopulations are always present in normal adult islets and have diverse gene expression profiles and distinct basal and glucose-stimulated insulin secretion. Importantly, the β cell subtype distribution is profoundly altered in type 2 diabetes. These data suggest that this antigenically defined β cell heterogeneity is functionally and likely medically relevant.


Potential therapeutic effect of Moroccan propolis in hyperglycemia, dyslipidemia, and hepatorenal dysfunction in diabetic rats.
The effect of propolis collected in Morocco on blood glucose, lipid profile, liver enzymes, and kidney function was investigated in control and diabetic rats. Antioxidant activity of propolis was evaluated with the use of DPPH, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS•+), ferric reducing power and total antioxidant activity assay. To study its effect in streptozotocin (STZ)-induced diabetes, the rats were divided into eight groups; four control and four diabetics. The animals received distilled water, glibenclamide, or propolis extract, 50 mg/kg/BW) or 100 mg/kg/b.wt, daily for 15 days. Blood glucose, triglyceride, lactic acid dehydrogenase, liver enzymes, creatinine, blood urea, lipid profile, and body weight were measured on day 15 after commencement of the treatment. Propolis has a strong antioxidant activity and high total flavonoids and polyphenols content. Glibenclamide and propolis have no significant effect on lipid parameters, and renal and hepatic function in non-diabetic rats. However, propolis or glibenclamide caused a significant lowering of blood glucose after a single administration and at day 15 after daily administration in diabetic rats (P<0.05). Both interventions significantly lowered lactic acid dehydrogenase, increased body weight, and ameliorated dyslipidemia and abnormal liver and kidney function caused by diabetes. The effect of propolis was dose-dependent and in a high dose it was more potent than glibenclamide. Propolis exhibited strong antihyperglycemic, antihyperlipidemic, and hepato-renal protective effects in diabetes, and significantly lowered the elevated lactic acid dehydrogenase. The study demonstrated for the first-time the effect of Moroccan propolis in diabetes and it will pave the way for clinical investigations.


Osteopontin gene expression determines spontaneous metastatic performance of orthotopic human breast cancer xenografts.
A major problem in the therapeutic management of cancer is the growth of metastases in distant organs, but the genes orchestrating the process need to be identified for the rational design of new treatment. Here, we provide decisive experimental evidence demonstrating the causal involvement of a specific gene, osteopontin (OPN), in the pathogenesis of metastasis by human breast cancer cells and implicating some of its probable partners. Stable long-term depletion, or up-regulation, of OPN gene expression in a matched, isogenic pair of human breast cancer cell lines of differing metastatic proficiency reproducibly changed their ability to colonize distant organs. OPN down-regulation was achieved by transduction of the metastatic line with a DNA construct encoding a small hairpin RNA in a vector labeled with red fluorescent protein and resulted in a marked reduction of metastatic load (P < 0.01). Up-regulation of OPN in the negligibly metastatic line, with a green fluorescent protein-marked retroviral vector containing OPN cDNA driven by a strong promoter, resulted in heavy colonization of the lungs and lymph nodes (P < 0.005). The reciprocal changes in behavior of these matched cell lines cross-corroborate each other. Concomitant changes were seen in the expression of other metastasis-related genes in both modulated lines. The data indicate that therapeutic targeting of tumor OPN molecules could reset metastatically relevant gene networks, resulting in clinical benefit.


Usefulness of the vitreous fluid analysis in the translational research of diabetic retinopathy.
Diabetic retinopathy (DR) is the major cause of acquired blindness in working-age adults. Current treatments for DR (laser photocoagulation, intravitreal corticosteroids, intravitreal antivascular endothelial growth factor (VEGF) agents, and vitreo-retinal surgery) are applicable only at advanced stages of the disease and are associated with significant adverse effects. Therefore, new pharmacological treatments for the early stages of the disease are needed. Vitreous fluid obtained from diabetic patients undergoing vitreoretinal surgery is currently used to explore the events that are taking place in the retina for clinical research. However, several confounding factors such as vitreous haemorrhage and concentration of vitreous proteins should be considered in the analysis of the results. In this paper we will focus on the vitreous fluid as a tool for exploring the mediators of DR and in particular the molecules related to inflammatory pathways. In addition, their role in the pathogenesis of DR will be discussed. The usefulness of new technologies such as flow cytometry and proteomics in identifying new candidates involved in the inflammatory process that occurs in DR will be overviewed. Finally, a more personalized treatment based on vitreous fluid analysis aiming to reduce the burden associated with DR is suggested.


Enhancing radiosensitivity: targeting the DNA repair pathways.
Radiotherapy is very effective in local control of cancerous tumors, but its curative potential is often limited by intrinsic radioresistance of the tumor cells. Since DNA repair pathways remove radiation-induced DNA lesions and protect cells from lethality, these pathways represent potential therapeutic targets to radiosensitize tumors. In order to achieve a therapeutic gain, however, there must be a differential between tumor and normal cells that can be exploited to preferentially target the DNA repair of the tumor, while sparing surrounding normal tissues, and this has represented a significant challenge to progress. Nevertheless, recent advances in our understanding of DNA repair mechanisms and tumor biology, on both the biochemical and genetic levels, have identified molecular differentials that may increase tumor specificity. This mechanistic insight suggests new strategies for radiotherapeutic targeting of DNA repair. Some of these strategies are reviewed here, including synthetic lethal, replicative stress, cell cycle and hypoxia-based approaches. The example of PARP1 inhibitor use in BRCA1 and 2 mutated breast cancer therapy is discussed, and future directions and challenges are explored.


Continuous and discretized pursuit learning schemes: various algorithms and their comparison.
A learning automaton (LA) is an automaton that interacts with a random environment, having as its goal the task of learning the optimal action based on its acquired experience. Many learning automata (LAs) have been proposed, with the class of estimator algorithms being among the fastest ones, Thathachar and Sastry, through the pursuit algorithm, introduced the concept of learning algorithms that pursue the current optimal action, following a reward-penalty learning philosophy. Later, Oommen and Lanctot extended the pursuit algorithm into the discretized world by presenting the discretized pursuit algorithm, based on a reward-inaction learning philosophy. In this paper we argue that the reward-penalty and reward-inaction learning paradigms in conjunction with the continuous and discrete models of computation, lead to four versions of pursuit learning automata. We contend that a scheme that merges the pursuit concept with the most recent response of the environment, permits the algorithm to utilize the LAs long-term and short-term perspectives of the environment. In this paper, we present all four resultant pursuit algorithms, prove the E-optimality of the newly introduced algorithms, and present a quantitative comparison between them.


Tenascin is accumulated along developing peripheral nerves and allows neurite outgrowth in vitro.
The extracellular matrix protein, tenascin, appears in a restricted pattern during organ morphogenesis. Here we studied the expression of tenascin along developing peripheral nerves in chick embryos and tested its activity as a substrate for cultured neurons. Motor axons grow out through the tenascin-rich, anterior part of the sclerotome. Shortly after, tenascin surrounds axon fascicles of ventral roots. At the limb levels, outgrowing axons accumulate in the tenascin-containing girdle region forming a plexus. In the limb, tenascin first appears in bracket-like structures that surround the precartilage cell condensations of the femur and humerus, respectively. These regions coincide with the channels along which axons first grow in from the girdle plexus to form the limb nerves. Later, the major tenascin staining is associated with the cartilage and tendon primordia, and not with the limb nerves. We used tenascin as a substrate for cultured neural explants and single cells in order to test for its function in neurite outgrowth. Dissociated embryonic neurons of various types attached to mixed polylysine/tenascin substrates and sprouted rapidly after a lag of several hours. Outgrowth was inhibited and neurites were detached by anti-tenascin antibodies. On substrates coated with tenascin alone, neurite outgrowth was achieved from 3 day spinal cord explants. Whereas growth cones were well spread and rapidly moving, the neurites were poorly attached, straight and rarely branched. We speculate that in vivo tenascin allows axonal outgrowth, but inhibits branching and supports fasciculation of newly formed axons.


The Influence of Drinking Buddies: A Longitudinal Investigation of Drinking Motivations and Drinking Behaviors in Emerging Adults.
Background: Heavy alcohol consumption and frequent alcohol use are associated with many adverse social and physical consequences. The different motivations underlying why people drink predict different patterns of alcohol consumption. A drinking buddy (i.e. a friend with whom a person drinks alcohol) influences a person's drinking via social learning, leading to escalations in drinking over time. Purpose: Few studies have investigated drinking motives among peers and none have studied whether the drinking motives of a drinking buddy can influence another person's drinking behavior; we sought to fill that gap. Method: Same-sex drinking buddies (N = 174; 66.1% female) were assessed once monthly for four months using self-report questionnaires. Participants were on average 18.66 years-old (SD = 1.17). Results: Indistinguishable actor-partner interdependence models using multilevel path analysis were conducted, with each drinking motive predicting drinking frequency and quantity, respectively. There were significant actor effects for social, enhancement, conformity, and coping motives; moreover, the enhancement, social, and coping-anxiety motives of the drinking buddy influenced the individual's drinking frequency across the four months of the study. Conversely, only the enhancement motives of the buddy predicted drinking quantity in the individual when averaged across time. Sex was not a significant moderator of these effects. Importance: When targeting risky drinking behavior in a therapeutic context, assessing and addressing a person's reasons for drinking, as well as their drinking buddy's reasons for drinking, may reduce the risk of escalations in either friend's drinking frequency over time.


Prevalence of Salmonella enterica, Listeria monocytogenes, and Escherichia coli virulence factors in bulk tank milk and in-line filters from U.S. dairies.
The zoonotic bacteria Salmonella enterica, Listeria monocytogenes, and Escherichia coli are known to infect dairy cows while not always causing clinical signs of disease. These pathogens are sometimes found in raw milk, and human disease outbreaks due to these organisms have been associated with the consumption of raw milk or raw milk products. Bulk tank milk (BTM) samples (536) and in-line milk filters (519) collected from dairy farms across the United States during the National Animal Health Monitoring System's Dairy 2007 study were analyzed by real-time PCR for the presence of S. enterica and pathogenic forms of E. coli and by culture techniques for the presence of L. monocytogenes. S. enterica was detected in samples from 28.1% of the dairy operations, primarily in milk filters. Salmonella was isolated from 36 of 75 PCR-positive BTM samples and 105 of 174 PCR-positive filter samples, and the isolates were serotyped. Cerro, Kentucky, Muenster, Anatum, and Newport were the most common serotypes. L. monocytogenes was isolated from 7.1% of the dairy operations, and the 1/2a complex was the most common serotype, followed by 1/2b and 4b (lineage 1). Shiga toxin genes were detected in enrichments from 15.2% of the BTM samples and from 51.0% of the filters by real-time PCR. In most cases, the cycle threshold values for the PCR indicated that toxigenic strains were not a major part of the enrichment populations. These data confirm those from earlier studies showing significant contamination of BTM by zoonotic bacterial pathogens and that the consumption of raw milk and raw milk products presents a health risk.


Short-term facilitation and depression in the cerebellum: some observations on wild-type and mutant rodents deficient in the extracellular matrix molecule tenascin C.
Short-term plasticity was studied on synapses to Purkinje cells (PC): paired-pulse facilitation in parallel fibers (PF) and paired-pulse depression in climbing fibers (CF). Both phenomena relate to synaptic strength. These forms of short-term plasticity were tested on cerebellar slices in rat by early postnatal synchronous stimulation of olivary neurons (i.e., CFs) with harmaline and by inhibition of a metabotropic glutamate receptor (mGluR) as well as in mice that were deficient in the extracellular matrix glycoprotein tenascin-C. Harmaline stimulation delayed the developmental competition between CF inputs and maintained multiple innervation. Paired-pulse depression of the CF-PC synapse after harmaline treatment was more expressed. However, paired-pulse facilitation in PF-PC synapses remained unchanged. Electrophysiological responses of postsynaptic mGluR1 in CF-PC synapses could be obtained only with AMPA receptors blocked and glutamate uptake impaired. The mGluR1-specific antagonist CPCCOEt suppressed the CF-mGluR EPSC in some PCs and potentiated it in other PCs. CF paired-pulse depression was not changed with CPCCOEt, thus excluding a presynaptic effect. The postsynaptic effect was underlined by CPCCOEt-induced rise in amplitude of EPSC and by a prolongation of its decay time. Tenascins are extracellular matrix glycoproteins that may restrict the regenerative capacity of the nervous tissue. Testing short-term presynaptic plasticity in tenascin-C-deficient mice showed that CF paired-pulse depression was less expressed while PF paired-pulse facilitation was augmented except in a group of cells where there was even depression. The results underline differences in forms of short-term plasticity with regard to susceptibility to diverse modulatory factors.


Thionated naphthalene diimides: tuneable chromophores for applications in photoactive dyads.
Varying the degree of thionation of a series of naphthalene diimide (NDI) and naphthalic imide (NI) phenothiazine dyad systems affords a systematic approach for tuning the system's donor-acceptor energy gap. Each dyad was compared to model NDI/NI systems and fully characterised through single crystal X-ray diffraction, NMR, cyclic voltammetry, electron paramagnetic resonance (EPR), transient absorption spectroscopy (TA), time-resolved infra-red spectroscopy (TRIR) and DFT. The measurements reveal that thionation increases both electron affinity of the NDI/NI acceptor dyad component and accessibility of the singly or doubly reduced states. Furthermore, FTIR and TA measurements show that excited state behaviour is greatly affected by thionation of the NDI and induces a decrease in the lifetime of the excited states formed upon the creation of charge-separated states.


Sample size requirements to detect the effect of a group of genetic variants in case-control studies.
Because common diseases are caused by complex interactions among many genetic variants along with environmental risk factors, very large sample sizes are usually needed to detect such effects in case-control studies. Nevertheless, many genetic variants act in well defined biologic systems or metabolic pathways. Therefore, a reasonable first step may be to detect the effect of a group of genetic variants before assessing specific variants. We present a simple method for determining approximate sample sizes required to detect the average joint effect of a group of genetic variants in a case-control study for multiplicative models. For a range of reasonable numbers of genetic variants, the sample size requirements for the test statistic proposed here are generally not larger than those needed for assessing marginal effects of individual variants and actually decline with increasing number of genetic variants in many situations considered in the group. When a significant effect of the group of genetic variants is detected, subsequent multiple tests could be conducted to detect which individual genetic variants and their combinations are associated with disease risk. When testing for an effect size in a group of genetic variants, one can use our global test described in this paper, because the sample size required to detect an effect size in the group is comparatively small. Our method could be viewed as a screening tool for assessing groups of genetic variants involved in pathogenesis and etiology of common complex human diseases.


The role of excitation-pattern cues in the detection of frequency shifts in bandpass-filtered complex tones.
One task intended to measure sensitivity to temporal fine structure (TFS) involves the discrimination of a harmonic complex tone from a tone in which all harmonics are shifted upwards by the same amount in hertz. Both tones are passed through a fixed bandpass filter centered on the high harmonics to reduce the availability of excitation-pattern cues and a background noise is used to mask combination tones. The role of frequency selectivity in this "TFS1" task was investigated by varying level. Experiment 1 showed that listeners performed more poorly at a high level than at a low level. Experiment 2 included intermediate levels and showed that performance deteriorated for levels above about 57 dB sound pressure level. Experiment 3 estimated the magnitude of excitation-pattern cues from the variation in forward masking of a pure tone as a function of frequency shift in the complex tones. There was negligible variation, except for the lowest level used. The results indicate that the changes in excitation level at threshold for the TFS1 task would be too small to be usable. The results are consistent with the TFS1 task being performed using TFS cues, and with frequency selectivity having an indirect effect on performance via its influence on TFS cues.


Investigating bacterial population structure and dynamics in traditional koumiss from Inner Mongolia using single molecule real-time sequencing.
Koumiss is considered as a complete dairy product high in nutrients and with medicinal properties. The bacterial communities involved in production of koumiss play a crucial role in the fermentation cycle. To reveal bacterial biodiversity in koumiss and the dynamics of succession in bacterial populations during fermentation, 22 samples were collected from 5 sampling sites and the full length of the 16S ribosomal RNA genes sequenced using single molecule real-time sequencing technology. One hundred forty-eight species were identified from 82 bacterial genera and 8 phyla. These results suggested that the structural difference in the bacterial community could be attributed to geographical location. The most significant difference in bacterial composition occurred in samples from group D compared with other groups. The sampling location of group D was distant from the city and maintained the primitive local nomadic life. The dynamics of succession in bacterial communities showed that Lactobacillus helveticus increased in abundance from 0 to 9h and reached its peak at 9h and then decreased. In contrast, Enterococcus faecalis, Enterococcus durans, and Enterococcus casseliflavus increased gradually throughout the fermentation process, and reached a maximum after 24h.


Thermal anomaly mapping from night MODIS imagery of USA, a tool for environmental assessment.
A method is presented for elevation, latitude and longitude decorrelation stretch of multi-temporal MODIS MYD11C3 imagery (monthly average night land surface temperature (LST) across USA and Mexico). Multiple linear regression analysis of principal components images (PCAs) quantifies the variance explained by elevation (H), latitude (LAT), and longitude (LON). The multi-temporal LST imagery is reconstructed from the residual images and selected PCAs by taking into account the portion of variance that is not related to H, LAT, LON. The reconstructed imagery presents the magnitude the standardized LST value per pixel deviates from the H, LAT, LON predicted. LST anomaly is defined as a region that presents either positive or negative reconstructed LST value. The environmental assessment of USA indicated that only for the 25 % of the study area (Mississippi drainage basin), the LST is predicted by the H, LAT, LON. Regions with milled climatic pattern were identified in the West Coast while the coldest climatic pattern is observed for Mid USA. Positive season invariant LST anomalies are identified in SW (Arizona, Sierra Nevada, etc.) and NE USA.


DNA methylation is correlated with gene expression during early pregnancy in Bos taurus.
Coordinated regulation of endometrial gene expression is essential for successful pregnancy establishment. A nonreceptive uterine environment may be a key contributor to pregnancy loss, as the majority of pregnancy losses occur prior to embryo implantation. DNA methylation has been highlighted as a potential contributor in regulating early pregnancy events in the uterus. It was hypothesized that DNA methylation regulates expression of key genes in the uterus during pregnancy. The correlation between DNA methylation and gene expression was tested. Endometrial samples from fertile and subfertile dairy cow strains were obtained at day 17 of pregnancy or the reproductive cycle. Microarrays were used to characterize genome-wide DNA methylation profiles and data compared with previously published transcription profiles. 39% of DNA methylation probes assayed mapped to RefSeq genes with transcription measurements. Correlations among gene expression and DNA methylation were assessed, and the 1,000 most significant correlations used for subsequent analysis. Of these, 52% percent were negatively correlated with gene expression. When this gene list was compared with previously reported gene expression studies on the same tissues, 42% were differentially expressed when pregnant and cycling animals were compared, and 11% were differentially expressed when pregnant fertile and subfertile animals were compared. DNA methylation status was correlated with gene expression in several pathways implicated in early pregnancy events. Although these data do not provide direct evidence of a causative association between DNA methylation and gene expression, this study provides critical support for an effect of DNA methylation in early pregnancy events and highlights candidate genes for future studies.


Shedding new light on the Crab with polarized X-rays.
Strong magnetic fields, synchrotron emission, and Compton scattering are omnipresent in compact celestial X-ray sources. Emissions in the X-ray energy band are consequently expected to be linearly polarized. X-ray polarimetry provides a unique diagnostic to study the location and fundamental mechanisms behind emission processes. The polarization of emissions from a bright celestial X-ray source, the Crab, is reported here for the first time in the hard X-ray band (~20-160 keV). The Crab is a complex system consisting of a central pulsar, a diffuse pulsar wind nebula, as well as structures in the inner nebula including a jet and torus. Measurements are made by a purpose-built and calibrated polarimeter, PoGO+. The polarization vector is found to be aligned with the spin axis of the pulsar for a polarization fraction, PF = (20.9 ± 5.0)%. This is higher than that of the optical diffuse nebula, implying a more compact emission site, though not as compact as, e.g., the synchrotron knot. Contrary to measurements at higher energies, no significant temporal evolution of phase-integrated polarisation parameters is observed. The polarization parameters for the pulsar itself are measured for the first time in the X-ray energy band and are consistent with observations at optical wavelengths.


Effects of hyperthermia and cepharanthin on adriamycin accumulation with changes in extracellular pH.
We compared adriamycin (ADR) accumulation with the intensity of intracellular fluorescence of 3,3'-(di-n-hexyl)-2,2'-oxacarbocyanine iodide (NK-2280), an indicator of cell membrane potential, after hyperthermia and examined the effects of cepharanthin (CEP) on the accumulation of ADR and NK-2280 in wild and ADR-resistant strains of Ehrlich ascites tumour (EAT) cells. Among wild 0.2 and 1-microgram ADR-resistant strains of EAT cells, intracellular accumulation of both ADR and NK-2280 decreased with an increase of ADR resistance. Hyperthermia at 42 degrees C and CEP induced a marked increase in accumulation of both ADR and NK-2280 in the ADR-resistant strains of EAT cells. The increase in ADR accumulation by hyperthermia and CEP is possibly due to an increase in the cell membrane potential and the inhibition of ADR efflux by CEP respectively. In the wild strain of EAT cells, we also evaluated the effect of extracellular pH change on ADR accumulation and the cell membrane potential and their alteration by hyperthermia and CEP. ADR accumulation decreased as pH decreased, but the cell membrane potential was not appreciably affected by pH change so far examined. Hyperthermia alone or combined with CEP significantly increased ADR accumulation in a pH range from 6.2 to 7.6. Hyperthermia increased the accumulation of ADR in the tumour cells, although its pH dependency was not completely resolved. CEP enhanced ADR accumulation more markedly in ADR-resistant cells than in wild-type cells. Therefore, the combination of hyperthermia and CEP may be very effective to increase the ADR cytotoxicity to ADR-resistant tumours.


Beneficial Effects of Oral Lactobacillus on Pain Severity in Women Suffering from Endometriosis: A Pilot Placebo-Controlled Randomized Clinical Trial.
This study assessed the effects of a lactobacillus-based medication on pain intensity scores in women with endometriosis. The present randomized pilot placebo-controlled trial was done on eligible women who were surgically and pathologically diagnosed with endometriosis. Thirty-seven participants who had not received hormonal treatment in the last three months, were enrolled and randomized into LactoFem® and placebo groups. Lactobacillus capsules or placebo were administrated orally once a day for 8 weeks. Patients were assessed for pain severity using Visual Analogue Scale (VAS) scores for dysmenorrhea, dyspareunia and chronic pelvic pain at baseline and after 8 and 12 weeks post-intervention. Mean age of participants and mean body mass index (BMI) for the LactoFem® and control groups were comparable. All patients had stage 3 and 4 of the disease based on revised American fertility society (AFS) classification of endometriosis. Mean initial pain scores for dysmenorrhea, dyspareunia and chronic pelvic pain were 6.53 ± 2.88, 4.82 ± 3.76 and 4.19 ± 3.53, respectively in the LactoFem® group and 5.60 ± 2.06, 3.67 ± 2.64 and 2.88 ± 2.80, respectively for the control group; the two groups had comparable scores in this regard. There was more decrease in pain scores for both dysmenorrhea and the overall pain after 8 weeks of treatment in LactoFem® group compared to the control group. The scores for dysmenorrhea were 6.53 ± 2.88 and 5.60 ± 2.06 in the LactoFem® and control groups, respectively, before intervention but, after 8-week treatment, these values were 3.07 ± 2.49 and 4.47 ± 2.13 (P=0.018), respectively. The changes in overall pain score in the LactoFem® and control group during this period were 7.33 ± 7.00 and 4.11 ± 1.68, respectively (P=0.017). This study showed some beneficial effects of lactobacillus administration on endometriosis-related pain (Registration number: IRCT20150819023684N5).


Burden of paediatric respiratory syncytial virus disease and potential effect of different immunisation strategies: a modelling and cost-effectiveness analysis for England.
Vaccines and prophylactic antibodies against respiratory syncytial virus (RSV) are in development and likely to be available in the next 5-10 years. The most efficient way to use these products when they become available is an important consideration for public health decision makers. We performed a multivariate regression analysis to estimate the burden of RSV in children younger than 5 years in England (UK), a representative high-income temperate country, and used these results to assess the potential effect of different RSV immunisation strategies (targeting vaccination for infants, or pregnant women, or prophylactic antibodies for neonates). We did a cost-effectiveness analysis for these strategies, implemented either separately or concurrently, and assessed the effect of restricting vaccination to certain months of the year. We estimated that RSV is responsible for 12 primary care consultations (95% CI 11·9-12·1) and 0·9 admissions to hospital annually per 100 children younger than 5 years (95% CI 0·89-0·90), with the major burden occurring in infants younger than 6 months. The most cost-effective strategy was to selectively immunise all children born before the start of the RSV season (maximum price of £220 [95% uncertainty interval (UI) 208-232] per vaccine, for an incremental cost-effectiveness ratio of £20 000 per quality-adjusted life-year). The maximum price per fully protected person that should be paid for the infant, newborn, and maternal strategies without seasonal restrictions was £192 (95% UI 168-219), £81 (76-86), and £54 (51-57), respectively. Nearly double the number of primary care consultations, and nearly five times the number of admissions to hospital occurred with RSV compared with influenza. RSV vaccine and antibody strategies are likely to be cost-effective if they can be priced below around £200 per fully protected person. A seasonal vaccination strategy is likely to provide the most direct benefits. Herd effects might render a year-round infant vaccination strategy more appealing, although it is currently unclear whether such a programme would induce herd effects. UK National Institute for Health Research.


Increased rates of molecular evolution in an equatorial plant clade: an effect of environment or phylogenetic nonindependence?
A recent study of environmental effects on rates of molecular evolution in the plant subgenus Mearnsia shows that species occurring in more equatorial latitudes have higher rates of substitution in rDNA sequences as compared to their more southerly congeners (Wright et al. 2003). However, we believe that the statistical approach employed by Wright et al. (2003) insufficiently accounts for the phylogenetic nonindependence of the species examined, given that all six equatorial species of Mearnsia form a clade. To distinguish between the effect of latitude and that of phylogenetic nonindependence, we have employed a variety of comparative approaches that use independent contrasts to test for an effect of environment across this entire subgenus. We find very little evidence for an effect of latitude on rate of molecular evolution using these approaches and believe that the shared evolutionary history of the clade is a plausible explanation of the apparent rate difference between equatorial and subequatorial Mearnsia species.


Anorexia nervosa: a unified neurological perspective.
The roles of corticotrophin-releasing factor (CRF), opioid peptides, leptin and ghrelin in anorexia nervosa (AN) were discussed in this paper. CRF is the key mediator of the hypothalamo-pituitary-adrenal (HPA) axis and also acts at various other parts of the brain, such as the limbic system and the peripheral nervous system. CRF action is mediated through the CRF1 and CRF2 receptors, with both HPA axis-dependent and HPA axis-independent actions, where the latter shows nil involvement of the autonomic nervous system. CRF1 receptors mediate both the HPA axis-dependent and independent pathways through CRF, while the CRF2 receptors exclusively mediate the HPA axis-independent pathways through urocortin. Opioid peptides are involved in the adaptation and regulation of energy intake and utilization through reward-related behavior. Opioids play a role in the addictive component of AN, as described by the "auto-addiction opioids theory". Their interactions have demonstrated the psychological aspect of AN and have shown to prevent the functioning of the physiological homeostasis. Important opioids involved are β-lipotropin, β-endorphin and dynorphin, which interact with both µ and κ opioids receptors to regulate reward-mediated behavior and describe the higher incidence of AN seen in females. Moreover, ghrelin is known as the "hunger" hormone and helps stimulate growth hormone (GH) and hepatic insulin-like-growth-factor-1(IGF-1), maintaining anabolism and preserving a lean body mass. In AN, high levels of GH due to GH resistance along with low levels of IGF-1 are observed. Leptin plays a role in suppressing appetite through the inhibition of neuropeptide Y gene. Moreover, the CRF, opioid, leptin and ghrelin mechanisms operate collectively at the HPA axis and express the physiological and psychological components of AN. Fear conditioning is an intricate learning process occurring at the level of the hippocampus, amygdala, lateral septum and the dorsal raphe by involving three distinct pathways, the HPA axis-independent pathway, hypercortisolemia and ghrelin. Opioids mediate CRF through noradrenergic stimulation in association with the locus coeruleus. Furthermore, CRF's inhibitory effect on gonadotropin releasing hormone can be further explained by the direct relationship seen between CRF and opioids. Low levels of gonadotropin have been demonstrated in AN where only estrogen has shown to mediate energy intake. In addition, estrogen is involved in regulating µ receptor concentrations, but in turn both CRF and opioids regulate estrogen. Moreover, opioids and leptin are both an effect of AN, while many studies have demonstrated a causal relationship between CRF and anorexic behavior. Moreover, leptin, estrogen and ghrelin play a role as predictors of survival in starvation. Since both leptin and estrogen are associated with higher levels of bone marrow fat they represent a longer survival than those who favor the ghrelin pathway. Future studies should consider cohort studies involving prepubertal males and females with high CRF. This would help prevent the extrapolation of results from studies on mice and draw more meaningful conclusions in humans. Studies should also consider these mechanisms in post-AN patients, as well as look into what predisposes certain individuals to develop AN. Finally, due to its complex pathogenesis the treatment of AN should focus on both the pharmacological and behavioral perspectives.


Management of acute myocardial infarction. Pathophysiological considerations.
Development of improved techniques for assessing infarct size in intact experimental animals and in patients has made it possible to evaluate the effectiveness of selected interventions more objectively. It is becoming increasingly apparent that infarct size is an important determinant of prognosis. Experimentally, infarct size can be modified favorably under specified conditions by interventions designed to reduce myocardial oxygen requirements or increase myocardial energy supply during the early evolution of infarction. Preliminary clinical applications of these concepts have been encouraging. The time seems to be right for carefully controlled clinical investigations to determine whether techniques designed to protect ischemic myocardium will be effective in improving survival and reducing morbidity from acute myocardial infarction. On the basis of available evidence, it seems clear that appropriate management of patients with this disorder should include careful attention to the impact of any therapeutic modality on the heart itself. It does not suffice to treat patients with beta-adrenergic agonists simply because they may elevate systemic arterial blood pressure. Hemodynamics can not be the sole criteria in guiding the physician charged with the responsibility for patient management. Rather, he must consider the potential effects of any contemplated therapeutic modality on myocardial viability as well. The ultimate place if interventions designed to protect ischemic myocardium in the therapeutic armamentarium remains to be determined. However, judicious therapy should be designed to minimize myocardial oxygen requirements and maximize myocardial energy supply as well as to prevent criti l impairment of peripheral perfusion.


Hypochlorous acid and hydrogen peroxide-induced negative regulation of Salmonella enterica serovar Typhimurium ompW by the response regulator ArcA.
Hydrogen peroxide (H2O2) and hypochlorous acid (HOCl) are reactive oxygen species that are part of the oxidative burst encountered by Salmonella enterica serovar Typhimurium (S. Typhimurium) upon internalization by phagocytic cells. In order to survive, bacteria must sense these signals and modulate gene expression. Growing evidence indicates that the ArcAB two component system plays a role in the resistance to reactive oxygen species. We investigated the influx of H2O2 and HOCl through OmpW and the role of ArcAB in modulating its expression after exposure to both toxic compounds in S. Typhimurium. H2O2 and HOCl influx was determined both in vitro and in vivo. A S. Typhimurium ompW mutant strain (∆ompW) exposed to sub-lethal levels of H2O2 and HOCl showed a decreased influx of both compounds as compared to a wild type strain. Further evidence of H2O2 and HOCl diffusion through OmpW was obtained by using reconstituted proteoliposomes. We hypothesized that ompW expression should be negatively regulated upon exposure to H2O2 and HOCl to better exclude these compounds from the cell. As expected, qRT-PCR showed a negative regulation in a wild type strain treated with sub-lethal concentrations of these compounds. A bioinformatic analysis in search for potential negative regulators predicted the presence of three ArcA binding sites at the ompW promoter region. By electrophoretic mobility shift assay (EMSA) and using transcriptional fusions we demonstrated an interaction between ArcA and one site at the ompW promoter region. Moreover, qRT-PCR showed that the negative regulation observed in the wild type strain was lost in an arcA and in arcB mutant strains. OmpW allows the influx of H2O2 and HOCl and is negatively regulated by ArcA by direct interaction with the ompW promoter region upon exposure to both toxic compounds.


Composition of nocturnal hypoxaemic burden and its prognostic value for cardiovascular mortality in older community-dwelling men.
To investigate the composition of nocturnal hypoxaemic burden and its prognostic value for cardiovascular (CV) mortality in community-dwelling older men. We analysed overnight oximetry data from polysomnograms obtained in 2840 men from the Outcomes of Sleep Disorders in Older Men (MrOS Sleep) study (ClinicalTrials.gov Identifier: NCT00070681) to determine the number of acute episodic desaturations per hour (oxygen desaturation index, ODI) and time spent below 90% oxygen saturation (T90) attributed to acute desaturations (T90desaturation) and to non-specific drifts in oxygen saturation (T90non-specific), respectively, and their relationship with CV mortality. After 8.8 ± 2.7 years follow-up, 185 men (6.5%) died from CV disease. T90 [hazard ratio (HR) 1.21, P < 0.001], but not ODI (HR 1.13, P = 0.06), was significantly associated with CV death in univariate analysis. T90 remained significant when adjusting for potential confounders (HR 1.16, P = 0.004). Men with T90 > 12 min were at an elevated risk of CV mortality (HR 1.59; P = 0.006). Approximately 20.7 (5.7-48.5) percent of the variation in T90 could be attributed to non-specific drifts in oxygen saturation. T90desaturation and T90non-specific were individually associated with CV death but combining both variables did not improve the prediction. In community-dwelling older men, T90 is an independent predictor of CV mortality. T90 is not only a consequence of frank desaturations, but also reflects non-specific drifts in oxygen saturation, both contributing towards the association with CV death. Whether T90 can be used as a risk marker in the clinical setting and whether its reduction may constitute a treatment target warrants further study.


Antibiotics for treating scrub typhus.
Scrub typhus, an important cause of acute fever in Asia, is caused by Orientia tsutsugamushi, an obligate intracellular bacterium. Antibiotics currently used to treat scrub typhus include tetracyclines, chloramphenicol, macrolides, and rifampicin. To assess and compare the effects of different antibiotic regimens for treatment of scrub typhus. We searched the following databases up to 8 January 2018: the Cochrane Infectious Diseases Group specialized trials register; CENTRAL, in the Cochrane Library (2018, Issue 1); MEDLINE; Embase; LILACS; and the metaRegister of Controlled Trials (mRCT). We checked references and contacted study authors for additional data. We applied no language or date restrictions. Randomized controlled trials (RCTs) or quasi-RCTs comparing antibiotic regimens in people with the diagnosis of scrub typhus based on clinical symptoms and compatible laboratory tests (excluding the Weil-Felix test). For this update, two review authors re-extracted all data and assessed the certainty of evidence. We meta-analysed data to calculate risk ratios (RRs) for dichotomous outcomes when appropriate, and elsewhere tabulated data to facilitate narrative analysis. We included six RCTs and one quasi-RCT with 548 participants; they took place in the Asia-Pacific region: Korea (three trials), Malaysia (one trial), and Thailand (three trials). Only one trial included children younger than 15 years (N = 57). We judged five trials to be at high risk of performance and detection bias owing to inadequate blinding. Trials were heterogenous in terms of dosing of interventions and outcome measures. Across trials, treatment failure rates were low.Two trials compared doxycycline to tetracycline. For treatment failure, the difference between doxycycline and tetracycline is uncertain (very low-certainty evidence). Doxycycline compared to tetracycline may make little or no difference in resolution of fever within 48 hours (risk ratio (RR) 1.14, 95% confidence interval (CI) 0.90 to 1.44, 55 participants; one trial; low-certainty evidence) and in time to defervescence (116 participants; one trial; low-certainty evidence). We were unable to extract data for other outcomes.Three trials compared doxycycline versus macrolides. For most outcomes, including treatment failure, resolution of fever within 48 hours, time to defervescence, and serious adverse events, we are uncertain whether study results show a difference between doxycycline and macrolides (very low-certainty evidence). Macrolides compared to doxycycline may make little or no difference in the proportion of patients with resolution of fever within five days (RR 1.05, 95% CI 0.99 to 1.10; 185 participants; two trials; low-certainty evidence). Another trial compared azithromycin versus doxycycline or chloramphenicol in children, but we were not able to disaggregate date for the doxycycline/chloramphenicol group.One trial compared doxycycline versus rifampicin. For all outcomes, we are uncertain whether study results show a difference between doxycycline and rifampicin (very low-certainty evidence). Of note, this trial deviated from the protocol after three out of eight patients who had received doxycycline and rifampicin combination therapy experienced treatment failure.Across trials, mild gastrointestinal side effects appeared to be more common with doxycycline than with comparator drugs. Tetracycline, doxycycline, azithromycin, and rifampicin are effective treatment options for scrub typhus and have resulted in few treatment failures. Chloramphenicol also remains a treatment option, but we could not include this among direct comparisons in this review.Most available evidence is of low or very low certainty. For specific outcomes, some low-certainty evidence suggests there may be little or no difference between tetracycline, doxycycline, and azithromycin as treatment options. Given very low-certainty evidence for rifampicin and the risk of inducing resistance in undiagnosed tuberculosis, clinicians should not regard this as a first-line treatment option. Clinicians could consider rifampicin as a second-line treatment option after exclusion of active tuberculosis.Further research should consist of additional adequately powered trials of doxycycline versus azithromycin or other macrolides, trials of other candidate antibiotics including rifampicin, and trials of treatments for severe scrub typhus. Researchers should standardize diagnostic techniques and reporting of clinical outcomes to allow robust comparisons.


Metronomic administration of ibandronate and its anti-angiogenic effects in vitro.
Angiogenesis plays an essential role in the growth and metastatic development of tumours. Recent in vitro studies have reported bisphosphonates as having anti-angiogenic properties. They have been shown to inhibit proliferation, induce apoptosis and decrease capillary-like tube formation, but often the in vitro concentrations and dosing schedules used do not reflect drug pharmacokinetics or clinical dosing regimens. Human umbilical vein endothelial cells were exposed to physiologically relevant doses of the bisphosphonate ibandronate, mimicking the clinical administration of oral ibandronate (1 h daily dosing over 8 days at concentrations ranging from 1-10 microM). Cellular growth characteristics were then assessed. Low-dose ibandronate (1.25-2 microM) significantly reduced endothelial cell growth, while 2 microM ibandronate also significantly reduced capillary-like tube formation and increased apoptosis of endothelial cells compared to untreated cells. There was no significant difference in activity with doses above 2 microM. However, inhibiting bFGF stimulated cell growth increased VEGF expression. This work has demonstrated that repeated low-dose drug administration (metronomic therapy) of ibandronate has certain anti-angiogenic properties by inhibiting the stimulatory effects of bFGF. However targeting the inhibition of bFGF alone is unlikely to be a successful approach for completely inhibiting angiogenesis due to the interplay between bFGF and VEGF.


Use of Cysteine Trapping to Map Spatial Approximations between Residues Contributing to the Helix N-capping Motif of Secretin and Distinct Residues within Each of the Extracellular Loops of Its Receptor.
Amino-terminal regions of secretin-family peptides contain key determinants for biological activity and binding specificity, although the nature of interactions with receptors is unclear. A helix N-capping motif within this region has been postulated to directly contribute to agonist activity while also stabilizing formation of a helix extending toward the peptide carboxyl terminus and docking within the receptor amino terminus. We used cysteine trapping to systematically explore spatial approximations between cysteines replacing each residue in this motif of secretin (sec), Phe(6), Thr(7), and Leu(10), and cysteines incorporated into the extracellular face of the receptor. Each peptide was a full agonist for cAMP, but had a lower binding affinity than natural hormone. These bound to COS cells expressing 61 receptor constructs incorporating cysteines in every position along each extracellular loop (ECL) and adjacent parts of transmembrane (TM) segments. Patterns of covalent labeling were distinct for each probe, with Cys(6)-sec labeling multiple residues in the carboxyl-terminal half of ECL2 and throughout ECL3, Cys(7)-sec predominantly labeling only single residues in the carboxyl-terminal end of ECL2 and the amino-terminal end of ECL3, and Cys(10)-sec not efficiently labeling any of these residues. These spatial constraints were used to refine our model of secretin bound to its receptor, now bringing ECL3 above the amino terminus of the ligand and revealing possible charge-charge interactions between this part of secretin and receptor residues in TM5, TM6, ECL2, and ECL3, which can orient and stabilize the peptide-receptor complex. This was validated by testing predicted approximations by mutagenesis and residue-residue complementation studies.


Picosecond optical signal processing based on transmissive fiber Bragg gratings.
We report the experimental realization of an ultrafast (terahertz-bandwidth) linear optical signal processor, particularly a picosecond flat-top optical pulse shaper, based on a fiber Bragg grating (FBG) working in transmission. The used FBG design technique, based on a specially apodized linearly chirped FBG, enables the synthesis of readily feasible devices with processing bandwidths well in the terahertz range. The specific device reported here is successfully demonstrated for reshaping ultrashort (400 fs FWHM) optical Gaussian-like pulses into 2 ps flat-top pulses.


Note: High-precision microsphere sorting using velocity sedimentation.
Monodisperse populations of microspheres are desirable for a variety of research and industrial applications, but many desirable sizes and materials can be difficult to synthesize and have limited commercial availability. In this paper, we present an effective, straightforward, and low cost method for sorting polydisperse microspheres into many separate monodisperse samples. The basic approach is to use velocity sedimentation through a density gradient in a long vertical column, followed by carefully targeted extraction. We demonstrate this technique by reducing the coefficient of variation of melamine microspheres from 13% to 1%-4% and glass microspheres from 35% to 3%-8%. This simple and inexpensive method can be used to sort microspheres of many sizes and materials, and is easily scalable, opening the possibility of cheap, monodisperse microspheres.


Identification of lactate as a driving force for prostanoid transport by prostaglandin transporter PGT.
We previously characterized the prostaglandin (PG) transporter PGT as an exchanger in which [(3)H]PGE(2) influx is coupled to the efflux of a countersubstrate. Here, we cultured HeLa cells that stably expressed human PGT under conditions known to favor glycolysis (glucose as a carbon source) or oxidative phosphorylation (glutamine as a carbon source) and studied the effect on PGT-mediated [(3)H]PGE(2) influx. PGT-expressing cells grown in glutamine exhibited a 2- to 4-fold increase in [(3)H]PGE(2) influx compared with the antisense control, whereas cells grown in glucose exhibited a 14-fold increase. In the presence of 10 vs. 25 mM glucose during the uptake, there was a dose-dependent increment in [(3)H]PGE(2) influx. Cis inhibition of [(3)H]PGE(2) influx occurred with lactate at physiological concentrations (apparent K(m) = 48 +/- 12 mM). Preloading with lactate caused a dose-dependent trans stimulation of PGT-mediated [(3)H]PGE(2) uptake, and external lactate caused trans stimulation of PGT-mediated [(3)H]PGE(2) release. Together, these data are consistent with PGT-mediated PG-lactate exchange. Cells engaged in glycolysis would then be poised energetically for prostanoid uptake by means of PGT.


Phactr1 regulates Slack (KCNT1) channels via protein phosphatase 1 (PP1).
The Slack (KCNT1) gene encodes sodium-activated potassium channels that are abundantly expressed in the central nervous system. Human mutations alter the function of Slack channels, resulting in epilepsy and intellectual disability. Most of the disease-causing mutations are located in the extended cytoplasmic C-terminus of Slack channels and result in increased Slack current. Previous experiments have shown that the C-terminus of Slack channels binds a number of cytoplasmic signaling proteins. One of these is Phactr1, an actin-binding protein that recruits protein phosphatase 1 (PP1) to certain phosphoprotein substrates. Using co-immunoprecipitation, we found that Phactr1 is required to link the channels to actin. Using patch clamp recordings, we found that co-expression of Phactr1 with wild-type Slack channels reduces the current amplitude but has no effect on Slack channels in which a conserved PKC phosphorylation site (S407) that regulates the current amplitude has been mutated. Furthermore, a Phactr1 mutant that disrupts the binding of PP1 but not that of actin fails to alter Slack currents. Our data suggest that Phactr1 regulates the Slack by linking PP1 to the channel. Targeting Slack-Phactr1 interactions may therefore be helpful in developing the novel therapies for brain disorders associated with the malfunction of Slack channels.


Combined effects of royal jelly and environmental enrichment against stress-induced cognitive and behavioral alterations in male rats: behavioral and molecular studies.
Exposure to chronic stress has detrimental effects on cognitive and emotional processing. Also, the neuroprotective influences of environmental enrichment (EE) and royal jelly (RJ) have been indicated in previous studies. To our knowledge, to date, there are no studies about the synergistic effects of EE and RJ on cognitive changes induced by stress. Therefore, this study aimed to investigate the protective effects of RJ, and EE on anxiety-like behaviors, cognitive functions, and expression of hippocampal and also prefrontal cortex (PFC) brain-derived neurotrophic factor (BDNF) levels in stressed rats. By using restraint and cold temperature, rats were exposed to stressful situations and then subjected to treatment with RJ or/ and EE for 14 days. Stress induction was done 14 days before treatments by placing the rats in the restrainer under 4°C. Following the interventions, anxiety-like behaviors, novel object recognition memory (NORM), inhibitive avoidance performance, hippocampal, and PFC BDNF expression were examined. The plasma corticosterone level of all groups was also evaluated. Results showed increased plasma corticosterone levels, stress-induced deficits in the NORM and IA tests, and increased anxiety-like behaviors. EE and RJ improved these deficits with a decline in serum corticosterone and also increased BDNF levels in the hippocampus and PFC in stressed ones. The EE and the RJ prevented the detrimental effects of stress on anxiety-like behaviors and memory processes. These treatments can protect susceptible brain areas against chronic stress via improvement in behavioral and cognitive impairments through mediating BDNF expression.


Clinical features and management of schwannoma affecting the upper and lower lips.
Head and neck schwannomas comprise 25-40% of all schwannomas, with presentation on the lips as the rarest and most surgically complicated site for perioral tumors. A systematic literature review was conducted to include 21 cases of patients with schwannoma of the upper or lower lips. The majority of patients presented with a single, painless, well-encapsulated nodule on the upper or lower lips. The nodules were consistently slow-growing, with an average 29.3 months from symptom onset to clinical presentation. Most cases were complicated by profound cosmetic disfigurement as well as dysphagia, dysarthria, snoring, and/or sleep apnea. Overall, histological analysis was consistent with classic schwannoma, and all cases were treated via complete surgical excision, and for malignant tumors, additional therapy was utilized. All but four cases achieved full remission by final follow-up. Recurrence rate for benign lip schwannomas was 5.3%, which is remarkably different from the standard recurrence rate of 8-24% for benign peripheral schwannomas. Additionally, the proportion of malignant tumors was greater for lip schwannomas than other schwannomas. Lip schwannomas demonstrate different characteristics than schwannomas from other locations on the body, and these remarkable differences highlight significant implications for clinical practice. Complete excision is the primary mode of treatment with overall excellent postoperative prognosis and rare instances of recurrence. Given the rarity of this tumor, this review of available cases serves to comprehensively describe clinical presentation and surgical treatment approaches to upper and lower lip schwannomas.


Growth of nonnecrotic tumors in the presence and absence of inhibitors.
In this article a model for the evolution of a spherically symmetric, nonnecrotic tumor is presented. The effects of nutrients and inhibitors on the existence and stability of time-independent solutions are studied. With a single nutrient and no inhibitors present, the trivial solution, which corresponds to a state in which no tumor is present, persists for all parameter values, whereas the nontrivial solution, which corresponds to a tumor of finite size, exists for only a prescribed range of parameters, which corresponds to a balance between cell proliferation and cell death. Stability analysis, based on a two-timing method, suggests that, where it exists, the nontrivial solution is stable and the trivial solution unstable. Otherwise, the trivial solution is stable. Modification to these characteristic states brought about by the presence of different types of inhibitors are also investigated and shown to have significant effect. Implications of the model for the treatment of cancer are also discussed.


Chromatic properties of neurons in macaque MT.
We have studied the responses of MT neurons to moving gratings, spatially modulated in luminance and chromaticity. Most MT neurons responded briskly and with high contrast sensitivity to targets whose luminance was modulated, with or without added chromatic contrast. When luminance modulation was removed and only chromatic stimulation was used, the responses of all MT neurons were attenuated. Most were completely unresponsive to stimulation with targets whose modulation fell within a "null" plane in color space; these null planes varied from neuron to neuron, but all lay close to the plane of constant photometric luminance. For about a third of the neurons, there was no color direction in which responses were completely abolished; almost all of these neurons had a definite minimum response for chromatic modulation near the isoluminant plane. MT neurons that responded to isoluminant targets did so inconsistently and with poor contrast sensitivity, so that only intensely modulated targets were effective. Whereas the best thresholds of MT neurons for luminance targets are close to behavioral contrast threshold, the thresholds for isoluminant targets lie considerably above behavioral contrast threshold. Therefore, although some MT neurons do give responses to isoluminant targets, they are unlikely to be the source of the chromatic motion signals revealed behaviorally.


Antiretroviral therapy-associated toxicities in the resource-poor world: the challenge of a limited formulary.
Toxicities related to antiretroviral therapy make long-term adherence to therapy difficult for patients and present challenges to providers, especially those in the resource-poor world who work with a limited formulary. In resource-poor settings, where limited drug options are the rule, when and how to change therapy are especially difficult problems. Drugs such as stavudine and didanosine are associated with serious metabolic complications, such as lactic acidosis, pancreatitis, and peripheral neuropathy. Antiretroviral agents associated with fewer metabolic effects, such as tenofovir and abacavir, remain widely unavailable. Because the current formulary restrictions appear to be unlikely to change quickly, providers in resource-poor countries must be familiar with the common adverse events-including metabolic complications, hypersensitivity reactions, anemia, and liver enzyme abnormalities-and must understand how to manage them with what is locally available. Most importantly, to avoid drug toxicities, a larger formulary is needed in resource-poor settings, and this must be a high priority for policy makers and health care professionals involved in treating human immunodeficiency virus infection globally.


Hydrocarbon constrained peptides - understanding preorganisation and binding affinity.
The development of constrained peptides represents an emerging strategy to generate peptide based probes and hits for drug-discovery that address challenging protein-protein interactions (PPIs). In this manuscript we report on the use of a novel α-alkenylglycine derived amino acid to synthesise hydrocarbon constrained BH3-family sequences (BIM and BID). Our biophysical and structural analyses illustrate that whilst the introduction of the constraint increases the population of the bioactive α-helical conformation of the peptide in solution, it does not enhance the inhibitory potency against pro-apoptotic Bcl-xL and Mcl-1 PPIs. SPR analyses indicate binding occurs via an induced fit mechanism whilst X-ray analyses illustrate none of the key interactions between the helix and protein are disturbed. The behaviour derives from enthalpy-entropy compensation which may be considered in terms of the ground state energies of the unbound constrained and unconstrained peptides; this has implications for the design of preorganised peptides to target protein-protein interactions.


Integrated platform and API for electrophysiological data.
Recent advancements in technology and methodology have led to growing amounts of increasingly complex neuroscience data recorded from various species, modalities, and levels of study. The rapid data growth has made efficient data access and flexible, machine-readable data annotation a crucial requisite for neuroscientists. Clear and consistent annotation and organization of data is not only an important ingredient for reproducibility of results and re-use of data, but also essential for collaborative research and data sharing. In particular, efficient data management and interoperability requires a unified approach that integrates data and metadata and provides a common way of accessing this information. In this paper we describe GNData, a data management platform for neurophysiological data. GNData provides a storage system based on a data representation that is suitable to organize data and metadata from any electrophysiological experiment, with a functionality exposed via a common application programming interface (API). Data representation and API structure are compatible with existing approaches for data and metadata representation in neurophysiology. The API implementation is based on the Representational State Transfer (REST) pattern, which enables data access integration in software applications and facilitates the development of tools that communicate with the service. Client libraries that interact with the API provide direct data access from computing environments like Matlab or Python, enabling integration of data management into the scientist's experimental or analysis routines.


Diagnostic accuracy of pulmonary embolism rule-out criteria: a systematic review and meta-analysis.
To perform a systematic review and meta-analysis to define the diagnostic performance of pulmonary embolism rule-out criteria (PERC) in deferring the need for D-dimer testing to rule out pulmonary embolism in the emergency department (ED). We searched EMBASE, MEDLINE, Scopus, Web of Knowledge, and all the evidence-based medicine reviews that included the Cochrane Database of Systematic Reviews through August 14, 2011, and hand searched references in potentially eligible articles and conference proceedings of major emergency medicine organizations for the previous 2 years. We selected studies that reported diagnostic performance of PERC, reported original research, and were conducted in the ED, with no language restrictions. Two investigators independently identified eligible studies and extracted data. We used contingency tables to calculate sensitivity, specificity, and likelihood ratios. We found 12 qualifying cohorts (studying 13,885 patients with 1,391 pulmonary embolism diagnoses), 10 prospective and 2 retrospective, from 6 countries. Pooled sensitivity, specificity, positive likelihood ratios, and negative likelihood ratios for 10 included studies were 0.97 (95% confidence interval [CI] 0.96 to 0.98), 0.23 (95% CI 0.22 to 0.24), 1.24 (95% CI 1.18 to 1.30), and 0.17 (95% CI 0.13 to 0.23), respectively. Significant heterogeneity was observed in specificity (I(2)=97.2%) and positive likelihood ratio (I(2)=84.2%). The existing literature suggests consistently high sensitivity and low but acceptable specificity of the PERC to rule out pulmonary embolism in patients with low pretest probability.


Changes in Traffic Crash Rates After Legalization of Marijuana: Results by Crash Severity.
The objective of this study was to estimate the effects of marijuana legalization and the subsequent onset of retail sales on injury and fatal traffic crash rates in the United States during the period 2009-2019. State-by-state quarterly crash rates per mile of travel were modeled as a function of time, unemployment rate, maximum posted speed limit, seat belt use rate, alcohol use rate, percent of miles driven on rural roads, and indicators of legalized recreational marijuana use and sales. Legalization of the recreational use of marijuana was associated with a 6.5% increase in injury crash rates and a 2.3% increase in fatal crash rates, but the subsequent onset of retail marijuana sales did not elicit additional substantial changes. Thus, the combined effect of legalization and retail sales was a 5.8% increase in injury crash rates and a 4.1% increase in fatal crash rates. Across states, the effects on injury crash rates ranged from a 7% decrease to an 18% increase. The effects on fatal crash rates ranged from a 10% decrease to a 4% increase. The estimated increases in injury and fatal crash rates after recreational marijuana legalization are consistent with earlier studies, but the effects varied across states. Because this is an early look at the time trends, researchers and policymakers need to continue monitoring the data.


A chiral BINOL-based Gemini amphiphilic gelator and its specific discrimination of native arginine by gelation in water.
A novel axially chiral cationic Gemini amphiphile gelator (S1) derived from (S)-BINOL has been synthesized and characterized by 1H NMR, 13C NMR, ESI-MS and FT-IR analyses. The critical micelle concentration (CMC) of S1 was determined to be 0.21 mM in water at room temperature. A transparent hydrogel with S1 at 43 mM was obtained at room temperature and characterized using various methods including SEM, CD, fluorescence, 1H NMR, FT-IR, and XRD. The results indicate that the hydrophobic effect of long alkyl chains, π-π stacking of naphthalene rings, and intermolecular hydrogen-bonding of the amide groups of S1 should be responsible for the hydrogel formation. Moreover, an 8.5 mM aqueous solution of S1 could gel by the addition of l-arginine, whereas it failed to gel in the presence of other 15 amino acids, respectively. It is suggested that S1 could discriminate native arginine by hydrogel formation, mainly due to the electrostatic interaction and hydrogen bonding effects between S1 and l-arginine molecules.


Efficacy of early diagnosis and treatment in women with a family history of breast cancer. European Familial Breast Cancer Collaborative Group.
Surveillance programmes for women at increased genetic risk of breast cancer are being established worldwide but little is known of their efficacy in early detection of cancers and hence reduction in mortality. Data were contributed from seven centres participating in the EU Demonstration Programme on Clinical Services for Familial Breast Cancer. All breast tumours (n = 161) detected prospectively, from the time of enrolment of women in a screening programme, were recorded. Analysis took account of age at diagnosis, whether tumours were screen-detected or not, their pathological stage and outcome by Kaplan-Meier survival plots. Mean age at diagnosis was 48.6 years. Overall, 75% of tumours were detected in the course of planned examinations. For women under age 50 at diagnosis, this figure was 68%. Eighteen percent were mammographically negative, (23% in patients under age 50). At first ("prevalence") round and at follow-up screening, 16% and 22% of tumours respectively were carcinoma in situ (CIS) while 27% and 22% respectively had evidence of nodal or distant spread (CaN+). Comparison of screen-detected and other tumours showed that the latter were more frequently mammogram-negative and CaN+. Overall five-year survival was 89% and five-year event-free survival 86%. Five-year event-free survival was 100% for CIS, 88% for invasive cancer without nodal or distant spread and 67% for CaN+. The majority of cancers arising in women at increased genetic risk of breast cancer can be detected by planned screening, even in those under age 50. Surveillance should include regular expert clinical examination and teaching of "breast awareness" as well as mammography. Attention to the logistics of screening programmes may improve still further the proportion of tumours that are screen-detected. The trend towards earlier pathological stage in tumours detected during follow-up rounds and the preliminary findings on survival analysis suggest that this approach will prove to be of long-term benefit for breast cancer families.


Evaluation of Burkholderia mallei ΔtonB Δhcp1 (CLH001) as a live attenuated vaccine in murine models of glanders and melioidosis.
Glanders caused by Burkholderia mallei is a re-emerging zoonotic disease affecting solipeds and humans. Furthermore, B. mallei is genetically related to B. pseudomallei, which is the causative agent of melioidosis. Both facultative intracellular bacteria are classified as tier 1 select biothreat agents. Our previous study with a B. mallei ΔtonB Δhcp1 (CLH001) live-attenuated vaccine demonstrated that it is attenuated, safe and protective against B. mallei wild-type strains in the susceptible BALB/c mouse model. In our current work, we evaluated the protective efficacy of CLH001 against glanders and melioidosis in the more disease-resistant C57BL/6 mouse strain. The humoral as well as cellular immune responses were also examined. We found that CLH001-immunized mice showed 100% survival against intranasal and aerosol challenge with B. mallei ATCC 23344. Moreover, this vaccine also afforded significant cross-protection against B. pseudomallei K96243, with low level bacterial burden detected in organs. Immunization with a prime and boost regimen of CLH001 induced significantly greater levels of total and subclasses of IgG, and generated antigen-specific splenocyte production of IFN-γ and IL-17A. Interestingly, protection induced by CLH001 is primarily dependent on humoral immunity, while CD4+ and CD8+ T cells played a less critical protective role. Our data indicate that CLH001 serves as an effective live attenuated vaccine to prevent glanders and melioidosis. The quantity and quality of antibody responses as well as improving cell-mediated immune responses following vaccination need to be further investigated prior to advancement to preclinical studies.


The effect of Rosa damascena essential oil on the amygdala electrical kindling seizures in rat.
We investigated the effect of Rosa damascena Mill, essential oil on the development of induced amygdala kindling seizures. Male Wistar rats were implanted with one tripolar and two monopolar electrodes in right basolateral amygdala and dura surface, respectively. The control group was injected solvent of essential oil and two experimental groups were injected 750 and 1000 mg kg(-1) of essential oil (ip), 30 min before a daily kindling stimulation. The number of stimulations required for the first appearance of seizure stages was significantly larger in two experimental groups than in control group. Mean after discharge duration was significantly different and essential oil reduced the increase of after discharge duration. Mean after discharge amplitude was also shorter in the groups treated with essential oil than in control group. Duration time for 5th stage of seizure at fully-kindled rats was significantly shorter in two experimental groups than control group. These results suggest that Rosa damascena essential oil significantly retarded the development of seizure stages and possesses the ability to counteract kindling acquisition. The flavonoids of Rosa damascena may act via GABAA receptors as previous studies have proposed for flavonoids of other medicinal plants. More detailed studies are recommended to define the effective component(s) of Rosa on different types of epilepsy.


Helping Chemists Discover New Antibiotics.
The world is facing a crisis in treating infectious diseases, with a scarcity of new antibiotics in development to treat the growing threat of drug-resistant "superbugs". We need new strategies to reinvigorate the antibiotic pipeline. In this Viewpoint we discuss one such approach, encouraging the community of synthetic chemists to participate in testing chemical diversity from their laboratories for antimicrobial potential. CO-ADD, the Community for Open Antimicrobial Drug Discovery, offers free screening against five bacteria and two fungi with follow up hit confirmation and validation, all with no strings attached.


Acceptability of oral versus rectal HIV preexposure prophylaxis among men who have sex with men and transgender women in Peru.
Oral preexposure prophylaxis (PrEP) with antiretrovirals (ARVs) is at the forefront of biomedical HIV prevention research, and ARVs are also being tested for rectal administration to target people practicing unprotected receptive anal intercourse (URAI) and at risk of HIV infection. This study assessed the acceptability of daily oral PrEP and rectal PrEP during URAI among men who have sex with men (MSM) and transgender women (TGW) in Peru. During the 2008 HIV sentinel surveillance survey conducted in 3 Peruvian cities (Lima, Iquitos, and Pucallpa), MSM and TGW reported being "versatile," "most of the time receptive," and "exclusively receptive" during anal sex behavior where surveyed on their acceptability of oral and rectal PrEP. Among 532 individuals, high acceptance of either oral (96.2%) or rectal (91.7%) PrEP products was reported. If both products were efficacious/available, 28.6% would prefer a pill, 57.3% a rectal lubricant, and 14.1% either. A trend toward higher acceptance was observed as receptive anal sex behavior exclusivity rose (P = .013). Being receptive most of the time (adjusted odds ratio [aOR]: 9.1, P = .01) and exclusively receptive (aOR: 7.5, P = .01), compared to being versatile, were independently associated with oral PrEP acceptability. A similar association was found with the acceptability of rectal formulations (aOR: 2.3, P = .07; and aOR: 2.5, P = .02; respectively). Oral and rectal PrEP were highly acceptable among Peruvian MSM and TGW, particularly among those at the highest HIV infection risk. These data can guide the implementation of PrEP programs in Peru and similar settings and populations.


Day Case Anterior Cruciate Ligament Reconstruction Surgery - A Study of Early Treatment Outcomes in a Regional Orthopaedic Centre.
Day-case anterior cruciate ligament reconstruction has the potential benefit of reduced hospital stay and reduced cost of care. The goal of this preliminary report was to compare the outcome of day-case arthroscopic anterior cruciate ligament reconstruction with those of in-patient care in terms of pain control and short-term functional outcome. This was a prospective comparative study involving patients who had anterior cruciate ligament reconstruction performed in our unit between January 2019 to July 2021 for isolated anterior cruciate ligament rupture. The patients were offered the option of in-patient and day-case anterior cruciate ligament reconstruction. All cases were isolated anterior cruciate ligament ruptures with no other ligament injury. A total of twenty-one-day case and twenty-five in-patient anterior cruciate ligament reconstruction were managed during the period of the study. The median numeric pain scores at day 2 and 7 in the day case group was 8.0 (IQR=2.0) and 5.0 (IQR= 3.0) respectively and in-patient group was 7.0 (IQR =1.5) and 4.0 (IQR= 2.0) respectively. The international knee documentation score (IKDC) at 6 months in the day case and in-patient groups were 68.6 (IQR= 9.3) and 67.2 (IQR= 25.0) respectively. The Mann-Whitney U test indicated that patients who had ACL reconstruction on in-patient care basis had statistically significant lower visual analogue scale pain scores on the second (z=-2.58, P = 0.01) and seventh (z=-3.41 P = 0.001) post-operative days compared to patients who had ACL reconstruction on day case basis. There was no statistically significant difference in the median IKDC scores of both groups at 6 months. The cost of care in the day case group was 40% lower than those of the in-patient group. Although the cost of care in the day case group appeared lower as compared to the in-patient group, the day case group had higher post-operative pain scores compared to the in-patient group. Although the post-operative functional scores were similar in both groups, this was not statistically significant.