Fruit and vegetable intake and head and neck cancer risk in a large United States prospective cohort study.
Squamous head and neck cancers include cancers of the oral cavity, pharynx and larynx are the sixth leading cause of cancer mortality worldwide, resulting in more than 350,000 deaths annually. Intake of fruit and vegetables may protect against head and neck cancer incidence, although few prospective studies have examined this association. We investigated this relation in 490,802 United States participants of the NIH-AARP Diet and Health cohort using Cox proportional hazard models adjusted for potential confounders. During 2,193,751 person years of follow-up from 1995/1996-2000, 787 participants were diagnosed with head and neck cancer. We found an inverse association between total fruit and vegetable intake and head and neck cancer risk (per serving/day/1,000 calories, Hazard Ratio, 95% Confidence interval: 0.94, 0.89-0.99). In models mutually adjusted for fruit and vegetable intake, the association was stronger for vegetables (fifth vs. first quintile: 0.65, 0.50-0.85) than for fruits (fifth vs. first quintile: 0.87, 0.68-1.11). When further subclassified into botanical groups, those in the highest tertile of leguminosae (dried beans, string beans and peas, 0.80, 0.67-0.96), rosaceae (apples, peach, nectarines, plums, pears and strawberries, 0.60, 0.49-0.73), solanaceae (peppers and tomatoes, 0.82, 0.69-0.98) and umbelliferae (carrots, 0.73, 0.60-0.89) had decreased risk of head and neck cancer, but no significant associations were seen for 9 other botanical groups. Results from this large prospective observational study are consistent with previous case-control studies and support the hypothesis that total fruit and vegetable intake is associated with reduced risk of head and neck cancer.


YAC complementation shows a requirement for Wt1 in the development of epicardium, adrenal gland and throughout nephrogenesis.
The Wilms' Tumour gene WT1 has important functions during development. Knock-out mice were shown to have defects in the urogenital system and to die at embryonic day E13.5, probably due to heart failure. Using a lacZ reporter gene inserted into a YAC construct, we demonstrate that WT1 is expressed in the early proepicardium, the epicardium and the subepicardial mesenchymal cells (SEMC). Lack of WT1 leads to severe defects in the epicardial layer and a concomitant absence of SEMCs, which explains the pericardial bleeding and subsequent embryonic death observed in Wt1 null embryos. We further show that a human-derived WT1 YAC construct is able to completely rescue heart defects, but only partially rescues defects in the urogenital system. Analysis of the observed hypoplastic kidneys demonstrate a continuous requirement for WT1 during nephrogenesis, in particular, in the formation of mature glomeruli. Finally, we show that the development of adrenal glands is also severely affected in partially rescued embryos. These data demonstrate a variety of new functions for WT1 and suggest a general requirement for this protein in the formation of organs derived from the intermediate mesoderm.


Genetic variation in the proximal promoter of ABC and SLC superfamilies: liver and kidney specific expression and promoter activity predict variation.
Membrane transporters play crucial roles in the cellular uptake and efflux of an array of small molecules including nutrients, environmental toxins, and many clinically used drugs. We hypothesized that common genetic variation in the proximal promoter regions of transporter genes contribute to observed variation in drug response. A total of 579 polymorphisms were identified in the proximal promoters (-250 to +50 bp) and flanking 5' sequence of 107 transporters in the ATP Binding Cassette (ABC) and Solute Carrier (SLC) superfamilies in 272 DNA samples from ethnically diverse populations. Many transporter promoters contained multiple common polymorphisms. Using a sliding window analysis, we observed that, on average, nucleotide diversity (pi) was lowest at approximately 300 bp upstream of the transcription start site, suggesting that this region may harbor important functional elements. The proximal promoters of transporters that were highly expressed in the liver had greater nucleotide diversity than those that were highly expressed in the kidney consistent with greater negative selective pressure on the promoters of kidney transporters. Twenty-one promoters were evaluated for activity using reporter assays. Greater nucleotide diversity was observed in promoters with strong activity compared to promoters with weak activity, suggesting that weak promoters are under more negative selective pressure than promoters with high activity. Collectively, these results suggest that the proximal promoter region of membrane transporters is rich in variation and that variants in these regions may play a role in interindividual variation in drug disposition and response.


Formation and distribution of nuclear pore complexes in interphase.
The possibility of nuclear pore formation in the interphase nucleus was investigated in control and phytohemagglutinin (PHA) stimulated lymphocytes by the freeze-etching technique. 48 hr after the addition of PHA, the newly formed blasts which had not as yet divided had at least twice the number of pores per nucleus as controls. This clearly demonstrates that in lymphocytes nuclear pore formation can take place during interphase. It has generally been assumed that the distribution of nuclear pore complexes in somatic animal cells is random. However, we have utilized freeze etched rat kidney cells and a computer program to evaluate pore distribution. We find a minimum pore center-to-center spacing of approximately 1300 A and multiples thereof with high frequency. This is strong evidence for a nonrandom distribution of nuclear pores. The nonrandomness may be related to an underlying chromosomal organization in interphase. Using three criteria for identifying prospective pore sites (membrane specialization, nonrandomness, and alteration of heterochromatin distribution), we have found forming pores in sectioned material from cultured human melanoma cells. While nuclear pore formation may take place in conjunction with reformation of the nuclear membrane, a mechanism also exists for their formation during interphase.


Alveostat, an alveolar PACO2 and PAO2 control system.
In the study of the physiological regulation of respiration through a control system model it is necessary to test the ventilatory response to various forcing functions of either the parrtial pressure of alveolar carbon dioxide (PACO2) or oxygen (PAO2). Since PACO2 and PAO2 are both functions of alveolar ventilation and metabolic rate, such a result cannot be obtained by merely changing the composition of the inspired gases without a feedback control. Thus a servomechanism is necessary. The input to the servomechanism is an instantaneous determination of PACO2 and PAO2. This is accomplished by using the criterion of equality of the exchange ratio in mean alveolar gas and mean expired gas. The servomechanism described has three essential characteristics: rapidity, accuracy, and stability. In experiments of step, ramp, and sinusoidal forcing functions, variations of PACO2 have been obtained without change in PAO2, and step variations of PAO2 have been obtained without change in PACO2.


Anti-PLA2R1 Antibodies Containing Sera Induce In Vitro Cytotoxicity Mediated by Complement Activation.
The phospholipase A2 receptor (PLA2R1) is the major autoantigen in idiopathic membranous nephropathy (MN). However, the pathogenic role of anti-PLA2R1 autoantibodies is unclear. Our aim was to evaluate the in vitro cytotoxicity of anti-PLA2R1 antibodies mediated by complement. Forty-eight patients with PLA2R1-related MN from the prospective cohort SOURIS were included. Anti-PLA2R1 titer, epitope profile, and anti-PLA2R1 IgG subclasses were characterized by ELISA. Cell cytotoxicity was evaluated by immunofluorescence in HEK293 cells overexpressing PLA2R1 incubated with patient or healthy donor sera in the presence or absence of rabbit complement or complement inhibitors. Mean cytotoxicity of anti-PLA2R1 sera for HEK293 cells overexpressing PLA2R1 was 2 ± 2%, which increased to 24 ± 6% after addition of rabbit complement (p < 0.001) (n = 48). GVB-EDTA, which inhibits all complement activation pathways, completely blocked cell cytotoxicity, whereas Mg-EGTA, which only inhibits the classical and lectin pathways, highly decreased suggesting a limited role of the alternative pathway. A higher diversity of IgG subclasses beyond IgG4 and high titer of total IgG anti-PLA2R1 were associated with increased cytotoxicity (p = 0.01 and p = 0.03 respectively). In a cohort of 37 patients treated with rituximab, high level of complement-mediated cytotoxicity was associated with less and delayed remission at month 6 after rituximab therapy (5/12 vs. 20/25 (p = 0.03) in 8.5 months ± 4.4 vs. 4.8 ± 4.0 (p = 0.02)). Kaplan-Meier analysis demonstrated that high level of cytotoxicity (≥40%) (p = 0.005), epitope spreading (defined by immunization beyond the immunodominant CysR domain) (p = 0.002), and high titer of anti-PLA2R1 total IgG (p = 0.01) were factors of poor renal prognosis. Anti-PLA2R1 antibodies containing sera can induce in vitro cytotoxicity mediated by complement activation, and the level of cytotoxicity increases with the diversity and the titer of anti-PLA2R1 IgG subclasses. These patients with high level of complement-mediated cytotoxicity could benefit from adjuvant therapy using complement inhibitor associated with rituximab to induce earlier remission and less podocyte injury.


Partial purification and characterization of human erythrocyte phosphoglycollate phosphatase.
Human erythrocytes contain a phosphatase that is highly specific for phosphoglycollate. It shows optimum pH of 6.7 and has Km 1 mM for phosphoglycollate. The molecular weight appears to be about 72000. The enzyme is a dimeric molecule having subunits of mol. wt. about 35000. It could be purified approx. 4000-fold up to a specific activity of 5.98 units/mg of protein. The activity of the enzyme is Mg2+-dependent. Co2+, and to a smaller extent Mn2+, may substitute for Mg2+. Half-maximum inhibition of the phosphatase by 5,5'-dithiobis-(2-nitrobenzoate), EDTA and NaF is obtained at 0.5 microM, 1 mM and 4 mM respectively. Moreover, it needs a univalent cation for optimum activity. Phosphoglycollate phosphatase is a cytoplasmic enzyme. Approx. 5% of its total activity is membrane-associated. This part of activity can be approx. 70% solubilized by freezing, thawing and treatment with 0.25% Triton X-100.


Risk of chronic dialysis and death following acute kidney injury.
Acute kidney injury frequently arises within an acute care hospitalization. Outcomes among acute kidney injury survivors following hospital discharge are poorly documented. We conducted a population-based cohort study between 1996 and 2006 of all adult patients in Ontario with acute kidney injury who did not require in-hospital dialysis, and who survived free of dialysis ≥30 days after discharge. Those with acute kidney injury (n=41,327) were matched 1:1 to patients without acute kidney injury during their index hospitalization. Matching was by age (±1 year), sex, history of chronic kidney disease, receipt of mechanical ventilation during the index hospitalization, and a propensity score for developing acute kidney injury. The primary outcome was subsequent need for chronic dialysis. The secondary outcomes were all-cause mortality and rehospitalization. Mean age was 70 years, and median follow-up was 2 years (maximum 10 years). The incidence of chronic dialysis was 1.78 per 100 person-years among those with acute kidney injury and 0.74 per 100 person-years among unaffected controls (adjusted hazard ratio [HR]; 2.70, 95% confidence interval [CI], 2.42-3.00). Rates also were higher for all-cause mortality (15.34 vs 14.51 per-100 person-years; adjusted HR 1.10; 95% CI, 1.07-1.13) and rehospitalization (44.93 vs 37.18 per 100 person-years; adjusted HR 1.21; 95% CI, 1.18-1.24). Even when acute dialysis is not required, survivors of acute kidney injury remain at higher risk of receipt of chronic dialysis thereafter. The absolute risk of death was more than 8 times the rate of chronic dialysis.


Triclosan resistance in Salmonella enterica serovar Typhimurium.
The aim of this study was to characterize the mechanisms of resistance to triclosan in Salmonella enterica serovar Typhimurium. Mutants resistant to triclosan were selected from nine S. enterica serovar Typhimurium strains. Mutants were characterized by genotyping, mutagenesis and complementation of fabI and analysis of efflux activity. Fitness of triclosan-resistant mutants was determined in vitro and in vivo. Three distinct resistance phenotypes were observed: low- (LoT), medium- (MeT) and high-level (HiT) with MICs of 4-8, 16-32 and >32 mg/L of triclosan, respectively, for inhibition. The genotype of fabI did not correlate with triclosan MIC. Artificial overexpression and mutagenesis of fabI in SL1344 each resulted in low-level triclosan resistance, indicating that FabI alone does not mediate high-level triclosan resistance in Salmonella Typhimurium. Active efflux of triclosan via AcrAB-TolC confers intrinsic resistance to triclosan as inactivation of acrB and tolC in wild-type strains and the triclosan-resistant mutants led to large decreases in triclosan resistance, which were reversed by complementation. Exemplars of each phenotype were evaluated for fitness in vivo; no fitness cost was seen and mutants colonized and persisted in chickens throughout a 28 day competitive index experiment. These data show that triclosan resistance can occur via distinct pathways in salmonella and that mutants selected after single exposure to triclosan are fit enough to compete with wild-type strains.


A Swedish Population-based Study of Adverse Birth Outcomes among Pregnant Women Treated with Buprenorphine or Methadone: Preliminary Findings.
Untreated opioid dependence in pregnant women is associated with adverse birth outcomes. Buprenorphine and methadone are options for opioid agonist medication-assisted treatment during pregnancy. The aim of this study was to describe adverse birth outcomes observed with buprenorphine or methadone treatment compared to the general population in Sweden. Pregnant women and their corresponding births during 2005-2011 were identified in the Swedish Medical Birth Register. Data on stillbirth, neonatal/infant death, mode of delivery, gestational age at birth, Apgar score, growth outcomes, neonatal abstinence syndrome, and congenital malformations were examined. Frequencies were compared using two-sided Fisher's exact tests. Unadjusted estimates of birth outcomes for women treated with buprenorphine or methadone were compared to the registered general population. A total of 746,257 pregnancies among 538,178 unique women resulted in 746,485 live births. Among the 194 women treated with buprenorphine (N = 176) or methadone (N = 52), no stillbirths or neonatal/infant deaths occurred. Neonatal abstinence syndrome developed in 23.3% and 38.5% of infants born to mothers treated with buprenorphine and methadone, respectively. The frequency of the selected adverse birth outcomes assessed in women treated with buprenorphine as compared to the general population was not significantly different. However, a significantly higher frequency of preterm birth and congenital malformations was observed in women treated with methadone as compared to the general population. Compared with the general population, methadone-treated women were significantly older than buprenorphine-treated women, and both treatment groups began prenatal care later, were more likely to smoke cigarettes, and did not cohabitate with the baby's father. An increased frequency of the selected adverse birth outcomes was not observed with buprenorphine treatment during pregnancy. Twofold increased frequency of preterm birth [2.21 (1.11, 4,41)] and congenital malformations [2.05 (1.08, 3.87)] was observed in the methadone group, which may be partly explained by older average maternal age and differences in other measured and unmeasured confounders.


Children's social goals and self-efficacy perceptions as influences on their responses to ambiguous provocation.
This study examined whether children who vary in their behavioral responses (aggression vs. withdrawal vs. problem solving) to ambiguous provocation but who are similar in their attributional processes differ in their social goals and self-efficacy perceptions. In response to 10 hypothetical situations involving ambiguous provocation, fourth- and fifth-grade students (n = 781) indicated whether or not the protagonist intended to cause the harm and reported how they would respond to the protagonist's action. Newly developed measures assessed children's situated social goals and self-efficacy perceptions. Results indicated that the aggressive, withdrawn, and problem-solving responders differed in their social goals and self-efficacy perceptions. The strength of the findings, compared to earlier work on children's goals and self-efficacy perceptions, suggests the importance of a situated social-cognitive assessment in which children's thoughts are measured in a specific kind of social situation and are related to their reported behavior in the same type of situation.


UK Renal Registry 16th annual report: chapter 15 epidemiology of reported infections amongst patients receiving dialysis for established renal Failure in England from May 2011 to April 2012: a joint report from Public Health England and the UK renal registry.
Infection remains one of the leading causes of mortality in established renal failure patients receiving renal replacement therapy (RRT). Since 2007, centres providing RRT in England have been asked to provide additional data on patients with methicillin resistant Staphylococcus aureus (MRSA) bacteraemia. Since 2011, the option to provide data on methicillin sensitive Stapylococcus aureus (MSSA) and Escherichia coli bacteraemia, as well as Clostridium difficile infection has also been available. Data were submitted to Public Health England by laboratories via HCAI-DCS including whether the patients were receiving dialysis. Individual renal centres then confirmed the record either directly via the database or after being contacted. Data were collected for the period of the 1st May 2011 to the 30th April 2012. There were 49 episodes of MRSA bacteraemia, an overall rate of 0.22 per 100 dialysis patients per year, representing a further year on year fall in MRSA rate. There were a higher number of MSSA episodes, 322 in total, with an overall rate of 1.15 per 100 dialysis patients per year. The number of episodes and overall rate of E. coli and C. difficile were 284 and 0.92 per 100 prevalent dialysis patients per year and 172 and 0.61 per 100 prevalent dialysis patients per year respectively. In each infection type the presence of a central venous catheter appeared to correlate with an elevated risk. Data are presented from one year of infections reported to PHE. The rate of MRSA bacteraemia episodes in England continues to fall. There was a higher rate of MSSA infections amongst renal dialysis patients. Findings from the first year of E. coli and C. difficile data collection are also reported. Future cycles will give us a further idea of the trend in incidences of these infections.


Promotion of incisional wound repair by human mesenchymal stem cell transplantation.
The purpose of this study was to determine the effect of transplanted human mesenchymal stem cells (hMSCs) on wound healing. In this model, full-thickness cutaneous wounds were created by incision in the skin of adult New Zealand white rabbits and treated by transplanted hMSCs into the wounds. Wound healing was evaluated by histological analysis and tensiometry over time. A total of 15 New Zealand white rabbits with 10 wounds per animal were examined in this study. Animals were treated with hMSCs and euthanised at 3, 7, 14, 21 and 80 days after manipulation. The hMSCs were labelled with a fluorescent dye (CM-DiI), suspended in phosphate-buffered saline and used to treat full-thickness incisional wounds in rabbit skin. Tensiometry and histology were used to characterise the wound-healing rate of the incisional wounds. These results showed that transplanted hMSCs significantly inhibited scar formation and increased the tensile strength of the wounds. Importantly, MSCs from genetically unrelated donors did not appear to induce an immunologic response. In conclusion, human mesenchymal stem cell therapy is a viable approach to significantly affect the course of normal cutaneous wound healing and significantly increase the tensile strength.


Nature of sub-band gap luminescent eigenmodes in a ZnO nanowire.
The emission spectrum of individual high-quality ZnO nanowires consists of a series of Fabry-Pérot-like eigenmodes that extend far below the band gap of ZnO. Spatially resolved luminescence spectroscopy shows that light is emitted predominantly at both wire ends, with identical spectra reflecting standing wave polariton eigenmodes. The intensity of the modes increases supralinearly with the excitation intensity, indicating that the mode population is governed by scattering among polaritons. Due to strong light-matter interaction, light emission from a ZnO nanowire is not dictated by the electronic band diagram of ZnO but depends also on the wire geometry and the excitation intensity. Delocalized polaritons provide a natural explanation for the pronounced subwavelength guiding in ZnO wires that has been reported previously.


Anti-inflammatory and antiarthritic activity of UNIM-301 (a polyherbal unani formulation) in Wistar rats.
UNIM-301 is a polyherbal formulation used in the Unani system of medicine for the treatment of joint pain and rheumatoid arthritis (RA). The objective was to evaluate the anti-inflammatory and antiarthritic activity of UNIM-301 in carrageenan-induced paw edema and complete Freund's Adjuvant (CFA) induced arthritis. The anti-inflammatory and antiarthritic activity of UNIM-301 was evaluated using carrageenan-induced paw edema and CFA induced animal arthritis models, respectively, in doses of 250, 500, and 1000 mg/kg body weight. Anti-inflammatory activity of UNIM-301 was evaluated using carrageenan-induced paw edema model using a digital plethysmometer. Anti-arthritic activity was evaluated using CFA induced arthritis, and joint sizes were measured at regular intervals using a micrometer screw gauge. Serum was collected and subjected to estimation of pro-inflammatory cytokine. Indomethacin 3 mg/kg body weight) was used as a standard drug in both the models. The acute and chronic toxicity study was carried out to evaluate the safety of the test drug. UNIM-301 treatment produced a dose-dependent reduction in paw edema and paw thickness in carrageenan-induced paw edema and CFA-induced arthritis, respectively, as compared to control. UNIM 301 also reduced the expression of pro-inflammatory mediator in a dose-dependent manner as compared to control. The result of the present study suggests that anti-inflammatory and anti-arthritic activity of UNIM-301, which might be accredited to inhibitory activity on pro-inflammatory cytokines to its various individual constituents.


Comparison of highway crash reporting in pakistan with the world health organization injury surveillance guidelines.
To compare the crash reporting system of National Highways & Motorways Police (NH&MP), Pakistan, with the World Health Organization (WHO) injury surveillance guidelines. Based on information collected from field observations, key informant interviews, and review of official documents, this note firstly describes the reporting system according to the components of a surveillance system. Then the reporting is compared with WHO criteria for designing and building an injury surveillance system and attributes of such a system. After a crash, a patrol officer communicates the information to the higher police authorities by wireless, fax, and on paper in the first 24 hours. Microcomputer Accident Analysis Package (MAAP) Performa filed by the officers are collected at a central location in the following 4 days, and reports are published biannually. Notable deficiencies in the reporting were nonidentification of stakeholders for data utilization and limited prospects of data recording process modification and its monitoring. Moreover, crash and injury definitions do not conform to international standards practiced elsewhere. NH&MP crash reporting needs to be simplified and standardized, and steps should be taken to improve its utilization for prevention purposes.


A canine model for studying endoleak after endovascular aneurysm repair.
The aim of this study was to create an animal model of endoleak after stent-graft placement for abdominal aortic aneurysm (AAA) in which a large aneurysmal sac would be preserved for the testing of techniques for its percutaneous occlusion. Infrarenal AAAs were created in nine dogs by anastomosis of an isolated segment of the inferior vena cava to the right side of the abdominal aorta in combination with a large anterior patch from the external jugular vein. One hour later, animals underwent percutaneous implantation of polytetrafluoroethylene-covered Z stent endografts with three 3-mm-diameter holes through the fabric. Aortograms were obtained before and after surgery, after endograft placement, and at the time of animal sacrifice at 1 week or 1, 2, 3, or 6 months. Pressures within the aorta and the aneurysm sac were recorded before animal sacrifice. Gross and histologic evaluations of the specimens were then carried out. Immediately after endograft placement, all nine animals had artificial type III endoleaks with angiographic filling of lumbar arteries and veins. One animal died of surgical complications within 2 days of surgery and is not included in our data analysis. One aneurysm ruptured at 1 week. At completion of the study, six endografts were patent and two were occluded. The aneurysm sac had enlarged by approximately 50% in seven animals. At follow-up, type I endoleak was present in three animals, type II endoleak was present in three, and the artificial type III endoleak was present in all six animals with patent endografts. The pressure differential between aorta and aneurysm sac was 36 mm Hg, with a mean aortic pressure of 87 mm Hg +/- 13.3 and a mean aneurysmal sac pressure of 51 mm Hg +/- 28.1. The aneurysmal sac exhibited early thrombus formation at 1 week, which progressed to complete thrombosis in 1-6 months. The model is technically feasible but would be useful in testing occlusive techniques for residual aneurysm sacs only in the acute phase after endograft placement. It would be not reliable for chronic evaluation because of rapidly progressive thrombosis in most aneurysm sacs and occasional complete thrombosis of the AAA and endograft.


Cerebral blood flow and ventilatory sensitivity to CO2 measured with the modified rebreathing method.
The ventilatory response to carbon dioxide (CO2) measured by modified rebreathing (SrVE) is closer to that measured by the steady-state method (SsVE) than is the response measured by Read's rebreathing method. Furthermore, the value estimated by the steady-state method depends upon the number of data points used to measure it. We planned to assess if these observations were also true for cerebral blood flow (CBF), as measured by steady-state (SsCBF) and modified rebreathing (SrCBF) tests. Six subjects undertook two protocols, one in the steady-state and one with modified rebreathing. SsVE depended upon the number of data points used to calculate it, and SsVE and SrVE were similar. However, this was not the case with SsCBF, and SsCBF was much higher than SrCBF. These findings are consistent with the notions that the specific CO2 stimulus differs for CBF control as compared with ventilation (VE) control, and that prior hypocapnia has an effect on CBF and VE for longer than the duration of the hypocapnia.


"Frozen evolution" of an RNA virus suggests accidental release as a potential cause of arbovirus re-emergence.
The mechanisms underlying virus emergence are rarely well understood, making the appearance of outbreaks largely unpredictable. Bluetongue virus serotype 8 (BTV-8), an arthropod-borne virus of ruminants, emerged in livestock in northern Europe in 2006, spreading to most European countries by 2009 and causing losses of billions of euros. Although the outbreak was successfully controlled through vaccination by early 2010, puzzlingly, a closely related BTV-8 strain re-emerged in France in 2015, triggering a second outbreak that is still ongoing. The origin of this virus and the mechanisms underlying its re-emergence are unknown. Here, we performed phylogenetic analyses of 164 whole BTV-8 genomes sampled throughout the two outbreaks. We demonstrate consistent clock-like virus evolution during both epizootics but found negligible evolutionary change between them. We estimate that the ancestor of the second outbreak dates from the height of the first outbreak in 2008. This implies that the virus had not been replicating for multiple years prior to its re-emergence in 2015. Given the absence of any known natural mechanism that could explain BTV-8 persistence over this long period without replication, we hypothesise that the second outbreak could have been initiated by accidental exposure of livestock to frozen material contaminated with virus from approximately 2008. Our work highlights new targets for pathogen surveillance programmes in livestock and illustrates the power of genomic epidemiology to identify pathways of infectious disease emergence.


Effect of cardiac arrest on cognitive impairment and hippocampal plasticity in middle-aged rats.
Cardiopulmonary arrest is a leading cause of death and disability in the United States that usually occurs in the aged population. Cardiac arrest (CA) induces global ischemia, disrupting global cerebral circulation that results in ischemic brain injury and leads to cognitive impairments in survivors. Ischemia-induced neuronal damage in the hippocampus following CA can result in the impairment of cognitive function including spatial memory. In the present study, we used a model of asphyxial CA (ACA) in nine month old male Fischer 344 rats to investigate cognitive and synaptic deficits following mild global cerebral ischemia. These experiments were performed with the goals of 1) establishing a model of CA in nine month old middle-aged rats; and 2) to test the hypothesis that learning and memory deficits develop following mild global cerebral ischemia in middle-aged rats. To test this hypothesis, spatial memory assays (Barnes circular platform maze and contextual fear conditioning) and field recordings (long-term potentiation and paired-pulse facilitation) were performed. We show that following ACA in nine month old middle-aged rats, there is significant impairment in spatial memory formation, paired-pulse facilitation n dysfunction, and a reduction in the number of non-compromised hippocampal Cornu Ammonis 1 and subiculum neurons. In conclusion, nine month old animals undergoing cardiac arrest have impaired survival, deficits in spatial memory formation, and synaptic dysfunction.


Repression of MHC class I gene promoter activity by two-exon Tat of HIV.
Major histocompatibility complex (MHC) class I molecules are the major receptors for viral peptides and serve as targets for specific cytotoxic T lymphocytes. Human immunodeficiency virus-type 1 (HIV-1) specifically decreased activity of an MHC class I gene promoter up to 12-fold. Repression was effected by the HIV-1 Tat protein derived from a spliced viral transcript (two-exon Tat). These studies define an activity for two-exon Tat distinct from that of one-exon Tat and suggest a mechanism whereby HIV-1-infected cells might be able to avoid immune surveillance, allowing the virus to persist in the infected host.


BRCA1 and BRCA2 proteins: roles in health and disease.
Between 5% and 10% of all breast cancer is hereditary, with patients having a strong family history of the disease. The remaining 90-95% of cases are classed as sporadic. Within the inherited group, 80-90% of cases are the result of germline mutations affecting two recently identified genes: BRCA1 and BRCA2. Since the sequencing of these genes, considerable research on the genetics of the mutation carriers has been performed, with less attention having been focused on the BRCA1 and BRCA2 proteins themselves. The structure and function of the protein products thus continues to hold mystery and might be the key to the full understanding of this complex disease.


Searching high and low: domestic dogs' understanding of solidity.
Physical reasoning appears central to understanding how the world works, suggesting adaptive function across the animal kingdom. However, conclusive evidence for inferential reasoning about physical objects is limited to primates. We systematically tested a central feature-understanding of solidity-in domestic dogs, by adapting a validated procedure (the shelf task) previously used to test children and non-human primates. Dogs watched a treat dropped into an apparatus with a shelf either present (treat landing on the shelf) or absent (treat landing on the bottom surface) and chose where to search for it (above or below the shelf). Across four studies (n = 64), we manipulated visual access to the treat trajectory and apparatus interior. Dogs correctly inferred the location of treats using physical cues when the shelf was present (Study 1), and learned rapidly when visual cues of continuity were limited (Study 2), and when the shelf was absent (Study 3). Dogs were at chance when the apparatus was fully occluded, and the presence and absence of the shelf varied across trials within subjects, and showed no evidence of learning (Study 4). The findings of these four studies suggest that dogs may be able to make some inferences using solidity and continuity and do not exhibit proximity or gravity biases. However, dogs did not always search correctly from Trial 1, and failed to search correctly when the rewarded location varied within-subjects, suggesting a role for learning, and possible limits to their ability to make inferences about physical objects.


Shedding of food-borne pathogens and microbiological carcass contamination in rabbits at slaughter.
To obtain microbiological data from rabbits at slaughter, 500 fecal samples and 500 carcasses samples were examined. All samples tested negative for Listeria and Salmonella. Campylobacter were detected in two fecal samples. Of the 500 fecal samples, 45.8% tested positive for eae (intimin), 1.2% for stx (Shiga toxin), and 1.8% for both eae and stx. By colony hybridization, 56 eae positive Escherichia coli strains were isolated. Among them, 27 strains (48.2%) were of the serotypes O178:H7 and O153:H7, whereas 15 strains (26.8%) belonged to a serogroup that has not yet been described (O(CB10681):H7). All strains possessed intimin beta1 and the translocated intimin receptor (tir) capable of being tyrosine phosphorylated. None of the strains harbored the genes for Shiga toxins, EAST1 (astA), bundlin (bfpA), or the EAF plasmid. Slaughter rabbits therefore constitute a reservoir for certain atypical enteropathogenic Escherichia coli. On rabbit carcasses, average total bacterial counts accounted for 3.3 log CFU cm(-2). Enterobacteriaceae and coagulase positive staphylococci (CPS) were detected on 118 (23.6%) and 153 (30.6%) carcasses, respectively. Enterobacteriaceae and CPS counts of positive samples were mainly <1.5 log CFU cm(-2). Among 153 selected CPS isolates, 98.7% were identified as Staphylococcus aureus. None of the 151 isolated strains harbored the gene for methicillin resistance (mecA). Genes for staphylococcal enterotoxins (SE) were detected in 102 strains. The combinations of seg and sei (53 strains) and sed, seg, sei, and sej (27 strains) dominated.


Relative and combined effects of socioeconomic status and diabetes on mortality: A nationwide cohort study.
Both low socioeconomic status (SES) and diabetes mellitus (DM) are important risk factors for mortality. However, little is known about their combined effects and relative contribution to the mortality risk.From a nationwide cohort provided by the National Health Insurance Service in Korea, 153,075 subjects who were over 30 years of age from 2003 to 2004 were followed-up until 2010. The SESs of the subjects in the DM and non-DM (NDM) groups were categorized into 3 groups (highest 30% as S1, middle 40% as S2, and lowest 30% as S3) based on the subjects' income levels.During the 7.9-year follow-up, 3933 deaths occurred. When the subjects were stratified into 6 groups by their socioeconomic and diabetes status, a linearly increasing pattern of the hazard ratio (HR) of mortality from the higher SES without diabetes group (NDM-S1, as a reference) to the lower SES with diabetes group (DM-S3; HR, 2.04, 95% confidence interval (CI), 1.80-2.36) was observed (P for trend < 0.001). Notably, subjects with DM in the highest SES group (DM-S1) had a significantly higher mortality risk than did non-DM subjects in the lowest SES group (NDM-S3). This pattern was maintained in cause-specific mortality but was more prominent in cardiovascular disease (CVD) and less prominent in cancer mortality. The association was not affected by gender; however, in individuals <60 years of age, the combined effects of SES and DM on mortality were more prominent (DM-S3; HR, 3.68, 95% CI, 2.95-4.60) than in those ≥60 years of age.Low SES and DM were major determinants of mortality and synergistically increased the risks of all-cause, CVD, and cancer mortality.


Seasonal variations in objectively assessed physical activity among people with COPD in two Nordic countries and Australia: a cross-sectional study.
Purpose: Seasons and weather conditions might influence participation in physical activity and contribute to differences between countries. This study aimed at investigating whether there were differences in physical activity levels between Norwegian, Danish and Australian people with chronic obstructive pulmonary disease (COPD), and establishing if any variations in physical activity were attributable to seasons. Patients and methods: A cross-sectional study where study subjects were people with COPD who participated in two separate clinical trials: the iTrain study (Norway, Denmark, and Australia) and the HomeBase study (Australia). Physical activity was objectively assessed with an activity monitor; variables were total energy expenditure, number of daily steps, awake sedentary time, light, and moderate-to-vigorous intensity physical activity. Differences in physical activity between countries and seasons were compared, with adjustment for disease severity. Results: In total, 168 participants were included from Norway (N=38), Denmark (N=36) and Australia (N=94). After controlling for disease severity, time spent in awake sedentary time was greater in Danish participants compared to the other countries (median 784 minutes/day [660-952] vs 775 minutes/day [626-877] for Norwegians vs 703 minutes/day [613-802] for Australians, P=0.013), whilst time spent in moderate to vigorous physical activity was lower (median 21 minutes/day [4-73] vs 30 minutes/day [7-93] for Norwegians vs 48 minutes/day [19-98] for Australians, P=0.024). Participants walked more during summer (median 3502 [1253-5407] steps/day) than in spring (median 2698 [1613-5207] steps/day), winter (median 2373 [1145-4206] steps/day) and autumn (median 1603 [738-4040] steps/day), regardless of geography. The median difference between summer and other seasons exceeded the minimal clinically important difference of 600 steps/day. However, the differences were not statistically significant (P=0.101). Conclusion: After controlling for disease severity, Danish participants spent more time in an awake sedentary state and less time in moderate to vigorous physical activity than their counterparts in Norway and Australia. People with COPD increased their physical activity in summer compared to other seasons. Weather conditions and seasonal variations may influence outcomes in clinical trials and health registries measuring physical activity over time, irrespective of the interventions delivered, and should be taken into account when interpreting results.


Reflection impulsivity in adolescent cannabis users: a comparison with alcohol-using and non-substance-using adolescents.
Reflection impulsivity-a failure to gather and evaluate information before making a decision-is a critical component of risk-taking and substance use behaviours, which are highly prevalent during adolescence. The Information Sampling Test was used to assess reflection impulsivity in 175 adolescents (mean age 18.3, range 16.5-20; 55% female)-48 cannabis users (2.3 years use, 10.8 days/month), 65 alcohol users, and 62 non-substance-using controls-recruited from a longitudinal cohort and from the general community and matched for education and IQ. Cannabis and alcohol users were matched on levels of alcohol consumption. Cannabis users sampled to the lowest degree of certainty before making a decision on the task. Group differences remained significant after controlling for relevant substance use and clinical confounds (e.g., anxiety, depressive symptoms, alcohol, and ecstasy use). Poor performance on multiple IST indices was associated with an earlier age of onset of regular cannabis use and greater duration of exposure to cannabis, after controlling for recent use. Alcohol users did not differ from controls on any IST measure. Exposure to cannabis during adolescence is associated with increased risky and impulsive decision making, with users adopting strategies with higher levels of uncertainty and inefficient utilisation of information. The young cannabis users did show sensitivity to losses, suggesting that greater impulsivity early in their drug using career is more evident when there is a lack of negative consequences. This provides a window of opportunity for intervention before the onset of cannabis dependence.


Attitude and willingness of high school students toward organ donation.
Public awareness of organ donation fundamentally affects the organ transplantation programs. This study was performed to assess the attitude and willingness of high school adolescents regarding organ donation. The study population consisted of 416 high school girls who were studying in four grades of three educational courses. Data were collected by a questionnaire and included demographic variables and attitude and willingness, which were assessed based on the Likert scale. The SPSS v.16 was used for data analysis. The mean age of the study subjects was 16.26 ± 1.06 years, 31% studied in grade-1, 27% in grade-2 (25% natural sciences, 27% mathematics and 48% humanities), 26% in grade-3 (30% natural sciences, 34% mathematics and 36% humanities) and 16% in pre-university stage (32% natural sciences, 42% mathematics and 26% humanities). The students had a highly positive attitude toward organ donation (mean score 4.2 ± 0.54). The greatest willingness for organ donation was concerning the kidney (88%) and heart (84%), followed by the liver (83.4%), pancreas (79.6%), cornea (67.8%) and skin (51%). Willingness for deceased as well as living organ donation was indicated by 92% and 47%, respectively, of the participants. Organ donation was considered acceptable only to relatives by 5% of the participants when the donors were deceased donors and by 16% of the participants when the donors were living donors; donation to all needy persons from deceased donors was accepted by 87% of the participants and from living donors by 31%. The purpose of donation was stated as lending help to others by 89% and progression of science by 40.2% of the participants. Willingness for organ donation from a deceased relative was declared by 63% of the students. There was significant positive correlation between willingness for organ donation and attitude (P <0.001). In addition, attitude and willingness had positive correlation with educational levels, age and educational courses. Our study suggests that Iranian adolescents have a great attitude and willingness toward organ donation, which reflects favorable knowledge of transplantation. Further research on public awareness in both genders and various age ranges is needed.


Electric field sensor array from cavity resonance between optical D-fiber and multiple slab waveguides.
We develop an electric field sensor array based on optical fiber interrogation with electro-optic crystals to measure high energy electromagnetic pulses. D-shaped optical fiber provides the platform for resonant coupling with multiple electro-optic crystals, allowing an array of sensing points on a single strand of optical fiber. Because of its small size, flexibility, and dielectric composition, this sensor array is suitable for performing electric-field analysis at multiple points within an electronic device. Using lithium niobate and potassium titanyl phosphate crystals, the sensor array is sensitive to fields as low as 100 V/m.


Unraveling the sequence-dependent polymorphic behavior of d(CpG) steps in B-DNA.
We have made a detailed study of one of the most surprising sources of polymorphism in B-DNA: the high twist/low twist (HT/LT) conformational change in the d(CpG) base pair step. Using extensive computations, complemented with database analysis, we were able to characterize the twist polymorphism in the d(CpG) step in all the possible tetranucleotide environment. We found that twist polymorphism is coupled with BI/BII transitions, and, quite surprisingly, with slide polymorphism in the neighboring step. Unexpectedly, the penetration of cations into the minor groove of the d(CpG) step seems to be the key element in promoting twist transitions. The tetranucleotide environment also plays an important role in the sequence-dependent d(CpG) polymorphism. In this connection, we have detected a previously unexplored intramolecular C-H···O hydrogen bond interaction that stabilizes the low twist state when 3'-purines flank the d(CpG) step. This work explains a coupled mechanism involving several apparently uncorrelated conformational transitions that has only been partially inferred by earlier experimental or theoretical studies. Our results provide a complete description of twist polymorphism in d(CpG) steps and a detailed picture of the molecular choreography associated with this conformational change.


Effect of confinement on proton-transfer reactions in water nanopools.
Proton transfer from the photoacid 8-hydroxy-1,3,6-pyrenetrisulfonic acid (HPTS) to water is studied in reverse micelles with ionic (AOT=sodium dioctyl sulfosuccinate) and non-ionic (BRIJ-30=polyoxyethylene(4)lauryl ether) surfactants. The dynamics are studied by probing the transient electronic absorption and transient vibrational absorption, both with sub-picosecond resolution. The reverse micelle sizes range from approximately 1.6 to 5.5 nm in diameter. For both surfactants it is found that the rate of proton transfer decreases with decreasing reverse micelle size, regardless of surfactant. In addition, for AOT reverse micelles, a fraction of the photoacid molecules exhibit non-radiative decay, preventing proton transfer.


Survival times of anomalous melt inclusions from element diffusion in olivine and chromite.
The chemical composition of basaltic magma erupted at the Earth's surface is the end product of a complex series of processes, beginning with partial melting and melt extraction from a mantle source and ending with fractional crystallization and crustal assimilation at lower pressures. It has been proposed that studying inclusions of melt trapped in early crystallizing phenocrysts such as Mg-rich olivine and chromite may help petrologists to see beyond the later-stage processes and back to the origin of the partial melts in the mantle. Melt inclusion suites often span a much greater compositional range than associated erupted lavas, and a significant minority of inclusions carry distinct compositions that have been claimed to sample melts from earlier stages of melt production, preserving separate contributions from mantle heterogeneities. This hypothesis is underpinned by the assumption that melt inclusions, once trapped, remain chemically isolated from the external magma for all elements except those that are compatible in the host minerals. Here we show that the fluxes of rare-earth elements through olivine and chromite by lattice diffusion are sufficiently rapid at magmatic temperatures to re-equilibrate completely the rare-earth-element patterns of trapped melt inclusions in times that are short compared to those estimated for the production and ascent of mantle-derived magma or for magma residence in the crust. Phenocryst-hosted melt inclusions with anomalous trace-element signatures must therefore form shortly before magma eruption and cooling. We conclude that the assumption of chemical isolation of incompatible elements in olivine- and chromite-hosted melt inclusions is not valid, and we call for re-evaluation of the popular interpretation that anomalous melt inclusions represent preserved samples of unmodified mantle melts.


Double dissociation of amygdala and hippocampal contributions to trace and delay fear conditioning.
A key finding in studies of the neurobiology of learning memory is that the amygdala is critically involved in Pavlovian fear conditioning. This is well established in delay-cued and contextual fear conditioning; however, surprisingly little is known of the role of the amygdala in trace conditioning. Trace fear conditioning, in which the CS and US are separated in time by a trace interval, requires the hippocampus and prefrontal cortex. It is possible that recruitment of cortical structures by trace conditioning alters the role of the amygdala compared to delay fear conditioning, where the CS and US overlap. To investigate this, we inactivated the amygdala of male C57BL/6 mice with GABA (A) agonist muscimol prior to 2-pairing trace or delay fear conditioning. Amygdala inactivation produced deficits in contextual and delay conditioning, but had no effect on trace conditioning. As controls, we demonstrate that dorsal hippocampal inactivation produced deficits in trace and contextual, but not delay fear conditioning. Further, pre- and post-training amygdala inactivation disrupted the contextual but the not cued component of trace conditioning, as did muscimol infusion prior to 1- or 4-pairing trace conditioning. These findings demonstrate that insertion of a temporal gap between the CS and US can generate amygdala-independent fear conditioning. We discuss the implications of this surprising finding for current models of the neural circuitry involved in fear conditioning.


Routine assistance to parents: effects on daily mood and other stressors.
The present study examined the association of providing assistance to older parents amid everyday circumstances and short-term psychological consequences for adult children providing assistance. We explored this association using 824 daily diary interviews of 119 adult children providing assistance in the National Study of Daily Experiences by using a left-censored random effects tobit regression model that accounted for the clustered data and floor effects in reported psychological distress. Psychological distress was higher on days adult children provided assistance to their parent (b = 0.88, p <.05) even after we controlled for situational variables such as time spent on daily paid work, time spent on leisure activities, and assistance provided to individuals other than parents. Demographic and psychosocial variables such as having resident children (b = 2.14, p <.01), less education (b = -0.54, p <.05), and neuroticism (b = 2.08, p <.05), also predicted daily psychological distress. Even after we controlled for within-person (daily situational variables) and between-person factors (background characteristics), the act of providing assistance itself had immediate associations with daily mood for helpers, particularly for those with fewer resources and greater demands on time. Feasibility and success of programs that provide respite and relief services to older adults and their children should be assessed in light of daily living.


The genome-scale metabolic network of Ectocarpus siliculosus (EctoGEM): a resource to study brown algal physiology and beyond.
Brown algae (stramenopiles) are key players in intertidal ecosystems, and represent a source of biomass with several industrial applications. Ectocarpus siliculosus is a model to study the biology of these organisms. Its genome has been sequenced and a number of post-genomic tools have been implemented. Based on this knowledge, we report the reconstruction and analysis of a genome-scale metabolic network for E. siliculosus, EctoGEM (http://ectogem.irisa.fr). This atlas of metabolic pathways consists of 1866 reactions and 2020 metabolites, and its construction was performed by means of an integrative computational approach for identifying metabolic pathways, gap filling and manual refinement. The capability of the network to produce biomass was validated by flux balance analysis. EctoGEM enabled the reannotation of 56 genes within the E. siliculosus genome, and shed light on the evolution of metabolic processes. For example, E. siliculosus has the potential to produce phenylalanine and tyrosine from prephenate and arogenate, but does not possess a phenylalanine hydroxylase, as is found in other stramenopiles. It also possesses the complete eukaryote molybdenum co-factor biosynthesis pathway, as well as a second molybdopterin synthase that was most likely acquired via horizontal gene transfer from cyanobacteria by a common ancestor of stramenopiles. EctoGEM represents an evolving community resource to gain deeper understanding of the biology of brown algae and the diversification of physiological processes. The integrative computational method applied for its reconstruction will be valuable to set up similar approaches for other organisms distant from biological benchmark models.


COVIDHunter: COVID-19 Pandemic Wave Prediction and Mitigation via Seasonality Aware Modeling.
Early detection and isolation of COVID-19 patients are essential for successful implementation of mitigation strategies and eventually curbing the disease spread. With a limited number of daily COVID-19 tests performed in every country, simulating the COVID-19 spread along with the potential effect of each mitigation strategy currently remains one of the most effective ways in managing the healthcare system and guiding policy-makers. We introduce COVIDHunter, a flexible and accurate COVID-19 outbreak simulation model that evaluates the current mitigation measures that are applied to a region, predicts COVID-19 statistics (the daily number of cases, hospitalizations, and deaths), and provides suggestions on what strength the upcoming mitigation measure should be. The key idea of COVIDHunter is to quantify the spread of COVID-19 in a geographical region by simulating the average number of new infections caused by an infected person considering the effect of external factors, such as environmental conditions (e.g., climate, temperature, humidity), different variants of concern, vaccination rate, and mitigation measures. Using Switzerland as a case study, COVIDHunter estimates that we are experiencing a deadly new wave that will peak on 26 January 2022, which is very similar in numbers to the wave we had in February 2020. The policy-makers have only one choice that is to increase the strength of the currently applied mitigation measures for 30 days. Unlike existing models, the COVIDHunter model accurately monitors and predicts the daily number of cases, hospitalizations, and deaths due to COVID-19. Our model is flexible to configure and simple to modify for modeling different scenarios under different environmental conditions and mitigation measures. We release the source code of the COVIDHunter implementation at https://github.com/CMU-SAFARI/COVIDHunter and show how to flexibly configure our model for any scenario and easily extend it for different measures and conditions than we account for.


Detection of Chlamydia trachomatis infection in urine samples from men and women by ligase chain reaction.
The suitability of urine specimens from women and men for the detection of Chlamydia trachomatis infection by a ligase chain reaction (LCR)-based assay with plasmid primers was examined with a group of patients attending a sexually transmitted disease clinic in Amsterdam, The Netherlands. Cervical specimens from 15 of 237 (6.3%) women tested positive for C. trachomatis by cell culture. Of the 25 (10.5%) female urine samples that tested positive by the plasmid-LCR assay, 13 were obtained from cervical culture-positive women. Nine of the 12 plasmid-LCR-positive urine samples from cervical culture-negative women were confirmed to be positive by a second LCR assay with primers based on chromosomal DNA. Urethral specimens from 24 of 258 (9.3%) men were positive for C. trachomatis infection by cell culture. Of the 25 (9.7%) urine samples that tested positive by plasmid-LCR, 20 were from culture-positive men. All five of the LCR-positive urine samples from culture-negative men were confirmed to be positive by the LCR with chromosomal DNA primers. Relative to cell culture, testing by plasmid-LCR analysis of male urine samples had a sensitivity of 83.3% and a specificity of 97.9%; after resolution of discordant samples, these values were 86.2 and 100%, respectively. In the study with women, the sensitivities of plasmid-LCR analysis of cervical and urine specimens in comparison with cervical cell culture were 93.3 and 86.7%, respectively. After resolution of discrepant samples, the sensitivities of the plasmid-LCR test for cervical swabs and female urine samples were 96.3 and 92.6%, respectively. These results indicate that the plasmid-LCR-based assay is a very reliable, sensitive, convenient test for the detection of C. trachomatis infection in female and male urine specimens.


Global and local depletion of ternary complex limits translational elongation.
The translation of genetic information according to the sequence of the mRNA template occurs with high accuracy and fidelity. Critical events in each single step of translation are selection of transfer RNA (tRNA), codon reading and tRNA-regeneration for a new cycle. We developed a model that accurately describes the dynamics of single elongation steps, thus providing a systematic insight into the sensitivity of the mRNA translation rate to dynamic environmental conditions. Alterations in the concentration of the aminoacylated tRNA can transiently stall the ribosomes during translation which results, as suggested by the model, in two outcomes: either stress-induced change in the tRNA availability triggers the premature termination of the translation and ribosomal dissociation, or extensive demand for one tRNA species results in a competition between frameshift to an aberrant open-reading frame and ribosomal drop-off. Using the bacterial Escherichia coli system, we experimentally draw parallels between these two possible mechanisms.


Characterization of NrnA homologs from Mycobacterium tuberculosis and Mycoplasma pneumoniae.
Processive RNases are unable to degrade efficiently very short oligonucleotides, and they are complemented by specific enzymes, nanoRNases, that assist in this process. We previously identified NrnA (YtqI) from Bacillus subtilis as a bifunctional protein with the ability to degrade nanoRNA (RNA oligos ≤5 nucleotides) and to dephosphorylate 3'-phosphoadenosine 5'-phosphate (pAp) to AMP. While the former activity is analogous to that of oligoribonuclease (Orn) from Escherichia coli, the latter corresponds to CysQ. NrnA homologs are widely present in bacterial and archaeal genomes. They are found preferably in genomes that lack Orn or CysQ homologs. Here, we characterize NrnA homologs from important human pathogens, Mpn140 from Mycoplasma pneumoniae, and Rv2837c from Mycobacterium tuberculosis. Like NrnA, these enzymes degrade nanoRNA and dephosphorylate pAp in vitro. However, they show dissimilar preferences for specific nanoRNA substrate lengths. Whereas NrnA prefers RNA 3-mers with a 10-fold higher specific activity compared to 5-mers, Rv2837c shows a preference for nanoRNA of a different length, namely, 2-mers. Mpn140 degrades Cy5-labeled nanoRNA substrates in vitro with activities varying within one order of magnitude as follows: 5-mer>4-mer>3-mer>2-mer. In agreement with these in vitro activities, both Rv2837c and Mpn140 can complement the lack of their functional counterparts in E. coli: CysQ and Orn. The NrnA homolog from Streptococcus mutans, SMU.1297, was previously shown to hydrolyze pAp and to complement an E. coli cysQ mutant. Here, we show that SMU.1297 can complement an E. coli orn(-) mutant, suggesting that having both pAp-phosphatase and nanoRNase activity is a common feature of NrnA homologs.


Nurses' reporting of suspect adverse drug reactions: a mixed-methods study.
To assess nurses' knowledge, attitudes and practices (KAP) towards spontaneous adverse drug reactions (ADRs) reporting. The mixed-method study was conducted following a quanti-qualitative sequential approach: a survey (using a KAP questionnaire) followed by a focus group was performed. In the quantitative findings, responders (570 hospital nurses) declared that they were unaware of the pharmacovigilance system (58.1%, n = 331); where to find the reporting form (63.5%, n = 362); how fill it in (71.6%, n = 408); to whom and how to send it (65.8%, n = 375). Only 11.1% (n = 63) reported ADRs. The qualitative phase supported the quantitative findings and provided new information about other factors that condition ADR reporting: misinterpretation of the meaning of "reporting", unawareness of nurses' autonomy in ADR reporting and fear of consequences after ADR reporting. Nurses are not fully aware of their role in ADR reporting. We recommend educational interventions and management changes.


Participation of Asian-American women in cancer treatment research: a pilot study.
Few Asian-American women participate in cancer treatment trials. In a pilot study to assess barriers to participation, we mailed surveys to 132 oncologists and interviewed 19 Asian-American women with cancer from Northern California. Forty-four oncologists responded. They reported as barriers language problems, lack of culturally relevant cancer information, and complex protocols. Most stated that they informed Asian-American women about treatment trials. Only four women interviewed knew about trials. Other patient-identified barriers were fear of side effects, language problems, competing needs, and fear of experimentation. Family decision making was a barrier for both oncologists and patients. Compared to non-Asian oncologists, more Asian oncologists have referred Asian-American women to industry trials and identified barriers similar to patients' reports. Our findings indicate that Asian-American women need to be informed about cancer treatment trials, linguistic barriers should be addressed, and future research should evaluate cultural barriers such as family decision making.


Calcium mobilization in human myeloid cells results in acquisition of individual dendritic cell-like characteristics through discrete signaling pathways.
We have shown previously that calcium ionophore (CI) treatment of various myeloid origin cells results in rapid acquisition of properties associated with mature, activated dendritic cells. These properties include increased CD83 and costimulatory molecule expression, tendencies to form dendritic processes, loss of CD14 expression by monocytes, and typically an enhanced capacity to sensitize T lymphocytes to Ag. We here analyze the intracellular signaling pathways by which CI induces acquisition of such properties. Thapsigargin, which raises intracellular Ca2+ levels by antagonizing its sequestration, induced immunophenotypic and morphologic changes that paralleled CI treatment. CI-induced activation was broadly attenuated by the Ca2+ chelating compound EGTA and by calmodulin antagonists trifluoperazine dimaleate and W-7. However, antagonists of signaling pathways downstream to calmodulin displayed more selective inhibitory effects. Calcineurin antagonists cyclosporin A and the FK-506 analogue, ascomycin, diminished costimulatory molecule and CD83 expression, as well as formation of dendritic processes in CI-treated myeloid cells, and strongly attenuated the T cell allosensitizing capacity of CI-treated HL-60 cells. These calcineurin antagonists displayed minimal effect on CI-induced CD14 down-regulation in monocytes. In contrast, the calmodulin-dependent protein kinase antagonists, K252a and KT5926, while displaying only modest effects on CI-induced costimulatory molecule and CD83 expression, strongly blocked CD14 down-regulation. These results are consistent with a Ca2+-dependent mechanism for CI-induced differentiation of myeloid cells, and indicate that multiple discrete signaling pathways downstream to calcium mobilization and calmodulin activation may be essential in regulating this process.


Ca2+ signals regulate mitochondrial metabolism by stimulating CREB-mediated expression of the mitochondrial Ca2+ uniporter gene MCU.
Cytosolic Ca2+ signals, generated through the coordinated translocation of Ca2+ across the plasma membrane (PM) and endoplasmic reticulum (ER) membrane, mediate diverse cellular responses. Mitochondrial Ca2+ is important for mitochondrial function, and when cytosolic Ca2+ concentration becomes too high, mitochondria function as cellular Ca2+ sinks. By measuring mitochondrial Ca2+ currents, we found that mitochondrial Ca2+ uptake was reduced in chicken DT40 B lymphocytes lacking either the ER-localized inositol trisphosphate receptor (IP3R), which releases Ca2+ from the ER, or Orai1 or STIM1, components of the PM-localized Ca2+ -permeable channel complex that mediates store-operated calcium entry (SOCE) in response to depletion of ER Ca2+ stores. The abundance of MCU, the pore-forming subunit of the mitochondrial Ca2+ uniporter, was reduced in cells deficient in IP3R, STIM1, or Orai1. Chromatin immunoprecipitation and promoter reporter analyses revealed that the Ca2+ -regulated transcription factor CREB (cyclic adenosine monophosphate response element-binding protein) directly bound the MCU promoter and stimulated expression. Lymphocytes deficient in IP3R, STIM1, or Orai1 exhibited altered mitochondrial metabolism, indicating that Ca2+ released from the ER and SOCE-mediated signals modulates mitochondrial function. Thus, our results showed that a transcriptional regulatory circuit involving Ca2+ -dependent activation of CREB controls the Ca2+ uptake capability of mitochondria and hence regulates mitochondrial metabolism.


Acute Myocardial Infarction in a Young Lady due to Vitamin B12 Deficiency Induced Hyperhomocysteinemia.
Hyper-homocysteinemia is a risk factor for coronary artery disease in young patients. A 32 years old female without any conventional risk factors except obesity presented with acute anterior wall myocardial infarction (MI). Her echocardiography showed anterior wall hypokinesia with moderate left ventricular dysfunction. Angiography showed tight stenosis of the proximal left anterior descending (LAD) and borderline lesion in left circumflex coronary artery (LCX). She underwent percutaneous coronary intervention (PCI) to LAD with good result. Her blood tests showed low vitamin B12, folate and serum iron levels and elevated serum homocysteine level. She was given folic acid and vitamin B12 and her homocysteine levels normalized. This case demonstrates that hyperhomocysteinemia caused by nutritional deficiency of vitamin co factors may lead to MI. Hyperhomocysteinemia should be considered in the evalauation of young people with MI, especially those without conventional risk factors.


Feeding practice and influence on selected blood parameters in show jumping horses competing in Switzerland.
The aim of this study was to compare the nutritional management of show jumping horses in practice with recommendations from the literature. Additionally, the effects of these feeding practices on several blood metabolic parameters before and after exercise were studied. Blood samples were collected in the field from 27 different horses at 71 trials on the level M1 to S2 show jumping competitions in Switzerland and questionnaires on feeding practice of the studied horses were evaluated. The questionnaires revealed that during training and on tournament days horses received on average 3.1 kg of concentrate per day (min. 2.0 kg, max. 6.6 kg) divided into two to three meals. The horses were fed on average 6.9 kg of roughage per day (min. 4.0 kg, max. 13.0 kg). Additionally, it was observed that the horses received the last meal on average 6 h 10 min (min. 1 h 50 min, max. 12 h 30 min) before the start of the first show jumping turn, respectively, 7 h 30 min (min 1 h 50 min, max. 13 h 0 min) before the second turn. Seven horses (35%) had access to hay waiting in the trailer between two turns. The statistical analysis revealed no significant influence of the concentrate feeding time point on lactate, triglyceride and insulin levels, but a significant influence on free fatty acids (FFA) and blood glucose concentrations. Roughage feeding of the show jumping horses 2-4 h prior to exercise revealed the most remarkable changes in blood parameters during the show jumping course. These results received under field conditions should be approved in future under standardized conditions.


War, peace, and fertility in Angola.
Using data from a nationally representative survey conducted in 1996, some two years after the end of a major outbreak of war, we examine the impact of war on the timing of recent births and war-related differences in reproductive preferences in Angola. We find evidence of a wartime drop and a postwar rebound in fertility, but these trends vary greatly, depending on the type and degree of exposure to war and on women's socioeconomic characteristics. At the same time, variations by parity are nonsignificant. In fertility preferences, the relative antinatalism of Angola's most modernized urban area stands out, but outside this area, differences between areas that were more and less affected by war are also noticeable. We offer interpretations of our findings and outline their implications for Angola's demographic future and demographic trends in similar settings.


The contribution of housing and neighbourhood conditions to educational inequalities in non-communicable diseases in Europe: findings from the European Social Survey (2014) special module on the social determinants of health.
Social gradients have been found across European populations, where less affluent groups are more often affected by poor housing and neighbourhood conditions. While poor housing and neighbourhood quality have been associated with a range of non-communicable diseases (NCDs), these conditions have rarely been applied to the examination of socioeconomic differences in NCDs. This study therefore asks 'to what extent does adjusting for poor housing and neighbourhood conditions reduce inequalities in NCDs among men and women in Europe'? Our analysis used pooled-data from 20 European countries for women (n= 12 794) and men (n= 11 974), aged 25–75, from round 7 of the European Social Survey. Fourteen NCDs were investigated: heart/circulatory problems, high blood pressure, back pain, arm/hand pain, foot/leg pain, allergies, breathing problems, stomach/digestion problems, skin conditions, diabetes, severe headaches, cancer, obesity and depression. We used binary logistic regression models, stratified by gender, and adjusted rate ratios to examine whether educational inequalities in NCDs were reduced after controlling for poor housing and neighbourhood quality. Overall, we find that adjusting for poor housing and neighbourhood quality reduces inequalities in NCDs. While reductions were relatively small for some NCDs–for high blood pressure, reductions were found in the range of 0–4.27% among women—for other conditions reductions were more considerable. Controlling for both housing and neighbourhood conditions for example, reduced inequalities by 16–24% for severe headaches and 14–30% for breathing problems. Social gradients in poor housing and neighbourhood quality could be an important contributor to educational inequalities in some NCDs.


Management of Hypothermia in Tactical Combat Casualty Care: TCCC Guideline Proposed Change 20-01 (June 2020).
As an outcome of combat injury and hemorrhagic shock, trauma-induced hypothermia (TIH) and the associated coagulopathy and acidosis result in significantly increased risk for death. In an effort to manage TIH, the Hypothermia Prevention and Management Kit™ (HPMK) was implemented in 2006 for battlefield casualties. Recent feedback from operational forces indicates that limitations exist in the HPMK to maintain thermal balance in cold environments, due to the lack of insulation. Consequently, based on lessons learned, some US Special Operations Forces are now upgrading the HPMK after short-term use (60 minutes) by adding insulation around the casualty during training in cold environments. Furthermore, new research indicates that the current HPMK, although better than no hypothermia protection, was ranked last in objective and subjective measures in volunteers when compared with commercial and user-assembled external warming enclosure systems. On the basis of these observations and research findings, the Committee on Tactical Combat Casualty Care decided to review the hypothermia prevention and management guidelines in 2018 and to update them on the basis of these facts and that no update has occurred in 14 years. Recommendations are made for minimal costs, low cube and weight solutions to create an insulated HPMK, or when the HPMK is not readily available, to create an improvised hypothermia (insulated) enclosure system.


A national outbreak of multi-resistant Salmonella enterica serovar Typhimurium definitive phage type (DT) 104 associated with consumption of lettuce.
Between 1 August and 15 September 2000, 361 cases of Salmonella enterica serotype Typhimurium definitive phage type (DT) 104, resistant to ampicillin, chloramphenicol, streptomycin, sulphonamides, spectinomycin and tetracycline (R-type ACSSuSpT), were identified in England and Wales residents. Molecular typing of 258 isolates of S. Typhimurium DT104 R-type ACSSuSpT showed that, although isolates were indistinguishable by pulsed-field gel electrophoresis, 67% (174/258) were characterized by a particular plasmid profile. A statistically significant association between illness and consumption of lettuce away from home was demonstrated (OR = 7.28; 95% CI=2.25-23.57; P=0.0006) in an unmatched case-control study. Environmental investigations revealed that a number of food outlets implicated in the outbreak had common suppliers of salad vegetables. No implicated foods were available for microbiological testing. An environmental audit of three farms that might have supplied salad vegetables to the implicated outlets did not reveal any unsafe agricultural practices. The complexity of the food supply chain and the lack of identifying markers on salad stuffs made tracking salad vegetables back to their origin extremely difficult in most instances. This has implications for public health since food hazard warnings and product withdrawal are contingent on accurate identification of the suspect product.


Spectral mapping of the third-order optical nonlinearity of glass-metal nanocomposites.
By tuning wavelengths of the femtosecond pump and probe pulses we mapped the nonlinear absorption of copper-glass nanocomposites within 520-620 nm range. At the pump intensity of 3 GW/cm(2), the induced transmission rise was as high as 20%. The imaginary part of the third-order optical susceptibility of the nanocomposites as a function of the probe wavelength reproduced well the spectral profile of the surface plasmon resonance in copper. In contrast, the imaginary part of the third-order optical susceptibility as a function of the pump wavelength did not reproduce the plasmon profile being wider than the latter.


Zebrafish hmx1 promotes retinogenesis.
Ocular development is controlled by a complex network of transcription factors, cell cycle regulators, and diffusible signaling molecules. Together, these molecules regulate cell proliferation, apoptosis and specify retinal fate. In the zebrafish (Danio rerio), hmx1 is a homeobox transcription factor implicated in eye and brain development. Hmx1 transcripts were detected in the nasal retina and lens as well as otic vesicles and pharyngeal arches by 24-32 hpf. Before this stage, transcripts were more uniformly expressed in the optic vesicle. Knockdown of hmx1 led to microphthalmia. Delayed withdrawal of retinal progenitors from the cell cycle resulting in retarded retinal differentiation was observed in morphant. The retina and brain also showed an increased cell death at 24 hpf. The polarized expression of hmx1 to the nasal part in the zebrafish retina strongly suggested an involvement in the nasal-temporal patterning. However, the key patterning genes tested so far were not regulated by hmx1. Altogether, these results suggest an important role for hmx1 in retinogenesis.


Clinical course correlates poorly with muscle pathology in nemaline myopathy.
To report pathologic findings in 124 Australian and North American cases of primary nemaline myopathy. Results of 164 muscle biopsies from 124 Australian and North American patients with primary nemaline myopathy were reviewed, including biopsies from 19 patients with nemaline myopathy due to alpha-actin (ACTA1) mutations and three with mutations in alpha-tropomyosin(SLOW) (TPM3). For each biopsy rod number per fiber, percentage of fibers with rods, fiber-type distribution of rods, and presence or absence of intranuclear rods were documented. Rods were present in all skeletal muscles and diagnosis was possible at all ages. Most biopsies contained nemaline bodies in more than 50% of fibers, although rods were seen only on electron microscopy in 10 patients. Rod numbers and localization correlated poorly with clinical severity. Frequent findings included internal nuclei and increased fiber size variation, type 1 fiber predominance and atrophy, and altered expression of fiber type specific proteins. Marked sarcomeric disruption, increased glycogen deposition, and intranuclear rods were associated with more severe clinical phenotypes. Serial biopsies showed progressive fiber size variation and increasing numbers of rods with time. Pathologic findings varied widely in families with multiple affected members. Very numerous nemaline bodies, glycogen accumulation, and marked sarcomeric disruption were common in nemaline myopathy associated with mutations in skeletal alpha-actin. Nemaline myopathy due to mutations in alpha-tropomyosin(SLOW) was characterized by preferential rod formation in, and atrophy of, type 1 fibers. Light microscopic features of nemaline myopathy correlate poorly with disease course. Electron microscopy may correlate better with disease severity and genotype.


Statistical Controversies in Reporting of Clinical Trials: Part 2 of a 4-Part Series on Statistics for Clinical Trials.
This paper tackles several statistical controversies that are commonly faced when reporting a major clinical trial. Topics covered include: multiplicity of data, interpreting secondary endpoints and composite endpoints, the value of covariate adjustment, the traumas of subgroup analysis, assessing individual benefits and risks, alternatives to analysis by intention to treat, interpreting surprise findings (good and bad), and the overall quality of clinical trial reports. All is put in the context of topical cardiology trial examples and is geared to help trialists steer a wise course in their statistical reporting, thereby giving readers a balanced account of trial findings.


Herbicides in ground water beneath Nebraska's Management Systems Evaluation Area.
Profiles of ground water pesticide concentrations beneath the Nebraska Management Systems Evaluation Area (MSEA) describe the effect of 20 yr of pesticide usage on ground water in the central Platte Valley of Nebraska. During the 6-yr (1991-1996) study, 14 pesticides and their transformation products were detected in 7848 ground water samples from the unconfined water table aquifer. Triazine and acetamide herbicides applied on the site and their transformation products had the highest frequencies of detection. Atrazine [6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4,-diamine] concentrations decreased with depth and ground water age determined with 3H/3He dating techniques. Assuming equivalent atrazine input during the past 20 yr, the measured average changes in concentration with depth (age) suggest an estimated half-life of >10 yr. Hydrolysis of atrazine and deethylatrazine (DEA; 2-chloro-4-amino-6-isopropylamino-s-triazine) to hydroxyatrazine [6-hydroxy-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine] appeared to be the major degradation route. Aqueous hydroxyatrazine concentrations are governed by sorption on the saturated sediments. Atrazine was detected in the confined Ogallala aquifer in ultra-trace concentrations (0.003 microg L(-1)); however, the possibility of introduction during reverse circulation drilling of these deep wells cannot be eliminated. In fall 1997 sampling, metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide] was detected in 57% of the 230 samples. Metolachlor oxanilic acid [(2-ethyl-6-methylphenyl)(2-methoxy-1-methylethyl) amino]oxo-acetic acid] was detected in most samples. In ground water profiles, concentrations of metolachlor ethane sulfonic acid [2-[(ethyl-6-methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxo-ethanesulfonic acid] exceeded those of deethylatrazine. Alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl)acetamide] was detected in <1% of the samples; however, alachlor ethane sulfonic acid [2-[(2,6-diethylphenyl)(methoxymethyl)amino]-2-oxoethanesulfonic acid] was present in most samples (63%) and was an indicator of past alachlor use.


Multiple Independent Origins of Apicomplexan-Like Parasites.
The apicomplexans are a group of obligate animal pathogens that include Plasmodium (malaria), Toxoplasma (toxoplasmosis), and Cryptosporidium (cryptosporidiosis) [1]. They are an extremely diverse and specious group but are nevertheless united by a distinctive suite of cytoskeletal and secretory structures related to infection, called the apical complex, which is used to recognize and gain entry into animal host cells. The apicomplexans are also known to have evolved from free-living photosynthetic ancestors and retain a relict plastid (the apicoplast), which is non-photosynthetic but houses a number of other essential metabolic pathways [2]. Their closest relatives include a mix of both photosynthetic algae (chromerids) and non-photosynthetic microbial predators (colpodellids) [3]. Genomic analyses of these free-living relatives have revealed a great deal about how the alga-parasite transition may have taken place, as well as origins of parasitism more generally [4]. Here, we show that, despite the surprisingly complex origin of apicomplexans from algae, this transition actually occurred at least three times independently. Using single-cell genomics and transcriptomics from diverse uncultivated parasites, we find that two genera previously classified within the Apicomplexa, Piridium and Platyproteum, form separately branching lineages in phylogenomic analyses. Both retain cryptic plastids with genomic and metabolic features convergent with apicomplexans. These findings suggest a predilection in this lineage for both the convergent loss of photosynthesis and transition to parasitism, resulting in multiple lineages of superficially similar animal parasites.


The Impact of Coconut Oil and Epigallocatechin Gallate on the Levels of IL-6, Anxiety and Disability in Multiple Sclerosis Patients.
Due to the inflammatory nature of multiple sclerosis (MS), interleukin 6 (IL-6) is high in blood levels, and it also increases the levels of anxiety related to functional disability. Epigallocatechin gallate (EGCG) decreases IL-6, which could be enhanced by the anti-inflammatory effect of high ketone bodies after administering coconut oil (both of which are an anxiolytic). Therefore, the aim of this study was to assess the impact of coconut oil and EGCG on the levels of IL-6, anxiety and functional disability in patients with MS. A pilot study was conducted for four months with 51 MS patients who were randomly divided into an intervention group and a control group. The intervention group received 800 mg of EGCG and 60 mL of coconut oil, and the control group was prescribed a placebo. Both groups followed the same isocaloric Mediterranean diet. State and trait anxiety were determined before and after the study by means of the State-Trait Anxiety Inventory (STAI). In addition, IL-6 in serum was measured using the ELISA technique and functional capacity was determined with the Expanded Disability Status Scale (EDSS) and the body mass index (BMI). State anxiety and functional capacity decreased in the intervention group and IL-6 decreased in both groups. EGCG and coconut oil improve state anxiety and functional capacity. In addition, a decrease in IL-6 is observed in patients with MS, possibly due to the antioxidant capacity of the Mediterranean diet and its impact on improving BMI.


Respiratory-induced coenzyme Q biosynthesis is regulated by a phosphorylation cycle of Cat5p/Coq7p.
CoQ(6) (coenzyme Q(6)) biosynthesis in yeast is a well-regulated process that requires the final conversion of the late intermediate DMQ(6) (demethoxy-CoQ(6)) into CoQ(6) in order to support respiratory metabolism in yeast. The gene CAT5/COQ7 encodes the Cat5/Coq7 protein that catalyses the hydroxylation step of DMQ(6) conversion into CoQ(6). In the present study, we demonstrated that yeast Coq7 recombinant protein purified in bacteria can be phosphorylated in vitro using commercial PKA (protein kinase A) or PKC (protein kinase C) at the predicted amino acids Ser(20), Ser(28) and Thr(32). The total absence of phosphorylation in a Coq7p version containing alanine instead of these phospho-amino acids, the high extent of phosphorylation produced and the saturated conditions maintained in the phosphorylation assay indicate that probably no other putative amino acids are phosphorylated in Coq7p. Results from in vitro assays have been corroborated using phosphorylation assays performed in purified mitochondria without external or commercial kinases. Coq7p remains phosphorylated in fermentative conditions and becomes dephosphorylated when respiratory metabolism is induced. The substitution of phosphorylated residues to alanine dramatically increases CoQ(6) levels (256%). Conversely, substitution with negatively charged residues decreases CoQ(6) content (57%). These modifications produced in Coq7p also alter the ratio between DMQ(6) and CoQ(6) itself, indicating that the Coq7p phosphorylation state is a regulatory mechanism for CoQ(6) synthesis.


Integrative characterization of the venom of the coral snake Micrurus dumerilii (Elapidae) from Colombia: Proteome, toxicity, and cross-neutralization by antivenom.
In Colombia, nearly 2.8% of the 4200 snakebite accidents recorded annually are inflicted by coral snakes (genus Micrurus). Micrurus dumerilii has a broad distribution in this country, especially in densely populated areas. The proteomic profile of its venom was here studied by a bottom-up approach combining RP-HPLC, SDS-PAGE and MALDI-TOF/TOF. Venom proteins were assigned to eleven families, the most abundant being phospholipases A2 (PLA2; 52.0%) and three-finger toxins (3FTx; 28.1%). This compositional profile shows that M. dumerilii venom belongs to the 'PLA2-rich' phenotype, in the recently proposed dichotomy for Micrurus venoms. Enzymatic and toxic venom activities correlated with protein family abundances. Whole venom induced a conspicuous myotoxic, cytotoxic and anticoagulant effect, and was mildly edematogenic and proteolytic, whereas it lacked hemorrhagic activity. Some 3FTxs and PLA2s reproduced the lethal effect of venom. A coral snake antivenom to Micrurus nigrocinctus demonstrated significant cross-recognition of M. dumerilii venom proteins, and accordingly, ability to neutralize its lethal effect. The combined compositional, functional, and immunological data here reported for M. dumerilii venom may contribute to a better understanding of these envenomings, and support the possible use of anti-M. nigrocinctus coral snake antivenom in their treatment. Coral snakes represent a highly diversified group of elapids in the New World, with nearly 70 species within the genus Micrurus. Owing to their scarce yields, the biochemical composition and toxic activities of coral snake venoms have been less well characterized than those of viperid species. In this work, an integrative view of the venom of M. dumerilii, a medically relevant coral snake from Colombia, was obtained by a combined proteomic, functional, and immunological approach. The venom contains proteins from at least eleven families, with a predominance of phospholipases A2 (PLA2), followed by three-finger toxins (3FTx). According to its compositional profile, M. dumerilii venom can be grouped with those of several Micrurus species from North and Central America that present a PLA2-predominant phenotype, to date it is the most southerly coral snake species to do so. Other coral snake species that a 'PLA2-rich' venom, M. dumerilii venom contains both components that form MitTx, a pain-inducing heterodimeric complex recently characterized from the venom of Micrurus tener, also present in Micrurus mosquitensis and M. nigrocinctus venoms. In addition to a lethal three-finger toxin, PLA2s participate in the toxicity of M. dumerilii venom, some of them displaying ability to induce cytolysis, muscle necrosis, and lethality to mice. An antivenom to M. nigrocinctus demonstrated significant cross-recognition of M. dumerilii venom proteins, and accordingly, ability to neutralize its lethal effect, being of potential therapeutic usefulness in these envenomings.


Deep brain stimulation suppresses pallidal low frequency activity in patients with phasic dystonic movements.
Deep brain stimulation of the globus pallidus internus alleviates involuntary movements in patients with dystonia. However, the mechanism is still not entirely understood. One hypothesis is that deep brain stimulation suppresses abnormally enhanced synchronized oscillatory activity within the motor cortico-basal ganglia network. Here, we explore deep brain stimulation-induced modulation of pathological low frequency (4-12 Hz) pallidal activity that has been described in local field potential recordings in patients with dystonia. Therefore, local field potentials were recorded from 16 hemispheres in 12 patients undergoing deep brain stimulation for severe dystonia using a specially designed amplifier allowing simultaneous high frequency stimulation at therapeutic parameter settings and local field potential recordings. For coherence analysis electroencephalographic activity (EEG) over motor areas and electromyographic activity (EMG) from affected neck muscles were recorded before and immediately after cessation of high frequency stimulation. High frequency stimulation led to a significant reduction of mean power in the 4-12 Hz band by 24.8 ± 7.0% in patients with predominantly phasic dystonia. A significant decrease of coherence between cortical EEG and pallidal local field potential activity in the 4-12 Hz range was revealed for the time period of 30 s after switching off high frequency stimulation. Coherence between EMG activity and pallidal activity was mainly found in patients with phasic dystonic movements where it was suppressed after high frequency stimulation. Our findings suggest that high frequency stimulation may suppress pathologically enhanced low frequency activity in patients with phasic dystonia. These dystonic features are the quickest to respond to high frequency stimulation and may thus directly relate to modulation of pathological basal ganglia activity, whereas improvement in tonic features may depend on long-term plastic changes within the motor network.


Tumor necrosis factor inhibitors are associated with reduced complement activation in spondylarthropathies: An observational study.
The complement system is involved in pathogenesis of cardiovascular disease, and might play a role in accelerated atherogenesis in spondylarthropathies (SpA). Hence, we examined complement activation in SpA, and its relationship to antirheumatic treatment, inflammatory and cardiovascular markers. From PSARA, a prospective observational study, we examined 51 SpA patients (31 psoriatic arthritis (PsA), and 20 ankylosing spondylitis (AS)), starting tumor necrosis factor (TNF) inhibitor alone (n = 25), combined with methotrexate (MTX) (n = 10), or MTX monotherapy (n = 16). Complement activation was determined by the soluble terminal complement complex (sC5b-9), inflammation by erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), and endothelial function by finger plethysmography (Endopat) at baseline, after 6 weeks and 6 months of treatment. SpA patients had sC5b-9 levels at (PsA) or above (AS) the upper limit of the estimated reference range. Median sC5b-9 levels decreased significantly from baseline to 6 weeks, with no significant difference between the AS and PsA group. Notably, a significant reduction in sC5b-9 was observed after administration of TNF inhibitor ± MTX, whereas no significant changes were observed in patients treated with MTX alone. Between 6 weeks and 6 months, sC5b-9 remained stable across all subgroups. Reduction in sC5b-9 was independently related to decreased ESR and CRP, and to increased high density cholesterol and total cholesterol. Reduction in sC5b-9 from baseline to 6 weeks was associated with improved EF in age and gender adjusted analyses. TNF-inhibition, but not MTX monotherapy, led to rapid and sustained reduction of complement activation in SpA. Thus, the observed decrease in cardiovascular morbidity in patients treated with TNF-inhibitors might be partly due to its beneficial effect on complement. Clinical Trials (NCT00902005), retrospectively registered on the 14th of May 2009.


Magnetic properties of segregated layers containing M(II)3(μ3-OH)2 (M = Co or Ni) diamond chains bridged by cis,cis,cis-1,2,4,5-cyclohexanetetracarboxylate.
We report the hydrothermal synthesis, X-ray crystal structures, and thermal, optical, and magnetic properties of two isostructural layered coordination polymers, [M(II)(5)(OH)(2)(chtc)(2)(H(2)O)(10)]·2H(2)O where M = Co (1Co) or Ni (2Ni) and chtc = cis,cis,cis-1,2,4,5-cyclohexanetetracarboxylate. The structures consist of segregated layers where each layer is formed of M(3)(OH)(2) diamond chains, involving edge- and corner-sharing octahedra, bridged by M(chtc)(2). In the space created by the chains and the bridges exists channels, which house the coordinated and noncoordinated water molecules, all hydrogen bonded to form a dodecamer, with a central cyclohexane chair shape ring. Interestingly, the water molecules of 2Ni are more difficult to remove than those of 1Co, but pyrolysis of the latter takes place at higher temperature. The magnetic properties are characterized by Néel transition to fully compensated antiferromagnets at 5.8 K (1Co) and 3.0 K (2Ni). The moments are easily reversed in an applied field of 150 Oe (1Co) and 300 Oe (2Ni) to a ferrimagnet with one uncompensated moment followed by a nonlinear increase to a saturation corresponding to a ferrimagnet with three uncompensated moments. Further, point charge calculations to estimate the weak bonding energies between the various types of interaction are reported.


Associations between self-rated psychosocial work conditions and musculoskeletal symptoms and signs. Stockholm MUSIC I Study Group.
The aim was to study the associations between self-rated psychosocial work conditions and the characteristics and location of musculoskeletal symptoms, signs, and syndromes. Perceived psychosocial work conditions were recorded in a cross-sectional study with 358 men and women in various occupations. Symptoms were recorded from the musculoskeletal system with a questionnaire, and signs were detected in a medical examination of all body regions. The analyses of statistical associations between the psychosocial factors and musculoskeletal disorders were performed with control for age, gender, and physical load. The most consistent and pronounced associations were mainly seen between poor psychosocial work conditions and coexisting symptoms and signs of the neck and back regions. Poor psychosocial work conditions were more consistently and strongly associated with signs of muscular (soft tissue) tenderness than with signs of tenderness in the joints, tendons, or muscular insertions or signs in nerve compression tests. Mainly low social support at work, but also high psychological demands and high job strain, were associated with such symptoms and signs, whereas decision latitude at work showed few associations with musculoskeletal disorders. Perceived poor psychosocial work conditions are statistically associated mainly with symptoms and signs of muscular tenderness in the central body regions. Studies on associations between psychosocial work conditions and musculoskeletal disorders should separate effect measures of different clinical signs and different body regions in order to avoid attenuation of the risk estimates.