Vegetation management with fire modifies peatland soil thermal regime.
Vegetation removal with fire can alter the thermal regime of the land surface, leading to significant changes in biogeochemistry (e.g. carbon cycling) and soil hydrology. In the UK, large expanses of carbon-rich upland environments are managed to encourage increased abundance of red grouse (Lagopus lagopus scotica) by rotational burning of shrub vegetation. To date, though, there has not been any consideration of whether prescribed vegetation burning on peatlands modifies the thermal regime of the soil mass in the years after fire. In this study thermal regime was monitored across 12 burned peatland soil plots over an 18-month period, with the aim of (i) quantifying thermal dynamics between burned plots of different ages (from <2 to 15 + years post burning), and (ii) developing statistical models to determine the magnitude of thermal change caused by vegetation management. Compared to plots burned 15 + years previously, plots recently burned (<2-4 years) showed higher mean, maximum and range of soil temperatures, and lower minima. Statistical models (generalised least square regression) were developed to predict daily mean and maximum soil temperature in plots burned 15 + years prior to the study. These models were then applied to predict temperatures of plots burned 2, 4 and 7 years previously, with significant deviations from predicted temperatures illustrating the magnitude of burn management effects. Temperatures measured in soil plots burned <2 years previously showed significant statistical disturbances from model predictions, reaching +6.2 °C for daily mean temperatures and +19.6 °C for daily maxima. Soil temperatures in plots burnt 7 years previously were most similar to plots burned 15 + years ago indicating the potential for soil temperatures to recover as vegetation regrows. Our findings that prescribed peatland vegetation burning alters soil thermal regime should provide an impetus for further research to understand the consequences of thermal regime change for carbon processing and release, and hydrological processes, in these peatlands.


De novo discovery of mutated driver pathways in cancer.
Next-generation DNA sequencing technologies are enabling genome-wide measurements of somatic mutations in large numbers of cancer patients. A major challenge in the interpretation of these data is to distinguish functional "driver mutations" important for cancer development from random "passenger mutations." A common approach for identifying driver mutations is to find genes that are mutated at significant frequency in a large cohort of cancer genomes. This approach is confounded by the observation that driver mutations target multiple cellular signaling and regulatory pathways. Thus, each cancer patient may exhibit a different combination of mutations that are sufficient to perturb these pathways. This mutational heterogeneity presents a problem for predicting driver mutations solely from their frequency of occurrence. We introduce two combinatorial properties, coverage and exclusivity, that distinguish driver pathways, or groups of genes containing driver mutations, from groups of genes with passenger mutations. We derive two algorithms, called Dendrix, to find driver pathways de novo from somatic mutation data. We apply Dendrix to analyze somatic mutation data from 623 genes in 188 lung adenocarcinoma patients, 601 genes in 84 glioblastoma patients, and 238 known mutations in 1000 patients with various cancers. In all data sets, we find groups of genes that are mutated in large subsets of patients and whose mutations are approximately exclusive. Our Dendrix algorithms scale to whole-genome analysis of thousands of patients and thus will prove useful for larger data sets to come from The Cancer Genome Atlas (TCGA) and other large-scale cancer genome sequencing projects.


ATP synthase deficiency due to TMEM70 mutation leads to ultrastructural mitochondrial degeneration and is amenable to treatment.
TMEM70 is involved in the biogenesis of mitochondrial ATP synthase and mutations in the TMEM70 gene impair oxidative phosphorylation. Herein, we report on pathology and treatment of ATP synthase deficiency in four siblings. A consanguineous family of Roma (Gipsy) ethnic origin gave birth to 6 children of which 4 were affected presenting with dysmorphic features, failure to thrive, cardiomyopathy, metabolic crises, and 3-methylglutaconic aciduria as clinical symptoms. Genetic testing revealed a homozygous mutation (c.317-2A>G) in the TMEM70 gene. While light microscopy was unremarkable, ultrastructural investigation of muscle tissue revealed accumulation of swollen degenerated mitochondria with lipid crystalloid inclusions, cristae aggregation, and exocytosis of mitochondrial material. Biochemical analysis of mitochondrial complexes showed an almost complete ATP synthase deficiency. Despite harbouring the same mutation, the clinical outcome in the four siblings was different. Two children died within 60 h after birth; the other two had recurrent life-threatening metabolic crises but were successfully managed with supplementation of anaplerotic amino acids, lipids, and symptomatic treatment during metabolic crisis. In summary, TMEM70 mutations can cause distinct ultrastructural mitochondrial degeneration and almost complete deficiency of ATP synthase but are still amenable to treatment.


Pragmatic recruitment of memantine as the capping group for the design of HDAC inhibitors: A preliminary attempt to unravel the enigma of glioblastoma.
Hurdled and marred by the notorious nature of glioblastomas (GBM) in terms of resistance to therapy and limited drug delivery into the brain, the anti-GBM drug pipeline is required to be loaded with mechanistically diverse agents. The consideration of HDAC inhibition as a prudent approach to circumvent the resistance issue in GBM spurred us to pragmatically design and synthesizes hydroxamic acids endowed with CNS penetrating ability. By virtue of the blood brain barrier permeability (BBB), memantine was envisioned as an appropriate CAP component for the construction of the HDAC inhibitors. Diverse linkers were stapled for the tetheration of the zinc binding motif with the CAP group to pinpoint an appropriate combination (CAP and linker) that could confer inhibitory preference to HDAC6 isoform (overexpressed in GBM). Resultantly, hydroxamic acid 16 was identified as a promising compound that elicited striking antiproliferative effects against Human U87MG GBM cells as well as TMZ-resistant GBM cells and P1S cells, a concurrent chemo radiotherapy (CCRT)-resistant/patient-derived glioma cell line mediated through preferential HDAC6 inhibition (IC50 = 5.42 nM). Furthermore, 16 exerted cell cycle arrest at G2 phase, induced apoptosis in GBM cells at high concentration and exhibited high BBB permeability. To add on, in-vivo study revealed that the administration of compound 16 prolonged the survival of TMZ-resistant U87MG inoculated orthotopic mice. Overall, the cumulative results indicate that 16 is a tractable CNS penetrant preferential HDAC6 inhibitor that might emerge as a potent weapon against GBM.


Serum antibody in Actinobacillus actinomycetemcomitans-infected patients with periodontal disease.
This study was designed to (i) delineate the characteristics of serum antibody responses to Actinobacillus actinomycetemcomitans in patients with periodontitis who are infected with A. actinomycetemcomitans; irrespective of disease classification; (ii) assess the relationship of the elevated antibody levels to colonization of the oral cavity by A. actinomycetemcomitans; and (iii) describe the serotype distribution of A. actinomycetemcomitans and antibodies to the microorganism in infected patients with various clinical classifications. To compare the levels of various isotype-specific antibodies to the different antigens, studies were performed that allowed quantitation of each isotype-specific antibody in a human reference standard. By using this reference standard, it was shown that the levels of immunoglobulin G (IgG), IgM, and IgA responses to A. actinomycetemcomitans were similar among the infected patients, irrespective of disease classification. Also, we demonstrated that the serum antibody response to serotype b was quantitatively greater in all isotypes. Our findings indicate that b was the most frequent A. actinomycetemcomitans serotype detected in the patients and appears to be capable of initiating a substantial serum IgG antibody response that may contain cross-reactive antibodies to other serotypes of A. actinomycetemcomitans. Generally, in cases in which the response to a single serotype was elevated, only that type of A. actinomycetemcomitans was detected in the plaque. Individuals exhibiting elevated antibodies to multiple serotypes were most consistently colonized by the serotype b microorganism. This study represents the first report detailing the distribution of IgG subclass antibodies to A. actinomycetemcomitans in periodontal disease. The results demonstrated that the primary responses of patients with periodontitis to A. actinomycetemcomitans were of the IgG1 and IgG3 subclasses, which is consistent with elicited responses to protein antigens. In contrast, the primary subclass response in normal subjects was limited to the IgG2 subclass and may represent broader cross-reactivity to polysaccharide antigens-lipopolysaccharide from the bacteria.


A field trial to evaluate a Mycoplasma hyopneumoniae vaccine: effects on lung lesions and growth rates in swine.
A killed Mycoplasma hyopneumoniae vaccine was evaluated in a single swine herd in which the farrowing barn and weaner rooms were on one Mycoplasma-free farm, while the growing and finishing barn was on a separate farm on which Mycoplasma was present. The study was carried out in a cohort of pigs born in a 12-week period. Pigs born in 6 of the 12 wk were vaccinated and the rest were left as controls. The vaccine was administered twice at approximately 3 and 6 wk of age. Carcass characteristics, lung lesions, and growth rates were recorded on 893, 390, and 220 pigs, respectively. The vaccine reduced the prevalence of pneumonic lesions in slaughter hogs from 69% to 36% (P < 0.001). It also appeared to reduce the prevalence of pleuritis from 20% to 13%, but the difference was only statistically significant at P = 0.07. The vaccine had no effects on carcass characteristics except that carcasses of vaccinated pigs were, on average, 1 kg heavier than those of nonvaccinated pigs, and a smaller percentage of vaccinated pigs were shipped "light" (carcass weight < 70 kg). Two methods were used to estimate the effect of the vaccine on growth rates (as measured by days to 80 kg carcass weight) resulting in estimates of 11 and 2 d reduction attributable to vaccination, respectively. The latter estimate was probably an underestimate for reasons discussed in the paper.


Rapamycin effects transcriptional programs in smooth muscle cells controlling proliferative and inflammatory properties.
Neointima formation, the leading cause of restenosis, is caused by proliferation of coronary artery smooth muscle cells (CASMCs) and is associated with infiltration by monocytes. Rapamycin inhibits neointima formation after stent implantation in humans. It reduces proliferation by its effects on mammalian target of rapamycin (mTOR) kinase. In this study, we investigated the expression of mTOR in human neointima and the effect of rapamycin on global transcriptional events controlling CASMC phenotype. In neointimal CASMCs, mTOR exhibited increased phosphorylation and was translocated to the nucleus compared with control. Comparative gene expression analysis of CASMCs treated with rapamycin (100 ng/ml) revealed down-regulation of the transcription factor E2F-1, a key regulator of G(1)/S-phase entry, and of various retinoblastoma protein/E2F-1-regulated genes. In addition, we found changes in the expression of genes associated with replication, apoptosis, and extracellular matrix formation. Furthermore, rapamycin decreased the gene expression of endothelial monocyte-activating polypeptide-II (EMAP-II). This decrease of EMAP-II expression was reflected in a reduced adhesiveness of CASMCs for monocytic cells. Addition of EMAP-II counteracted the antiadhesive effect of rapamycin. Therefore, EMAP-II may comprise a mechanism of rapamycin-mediated reduction of the proinflammatory activation of CASMCs. The effects reported here of rapamycin on the down-regulation of genes involved in cell cycle progression, apoptosis, proliferation, and extracellular matrix formation in CASMCs provide an explanation of how rapamycin reduces CASMC proliferation. In addition, rapamycin may contribute to a reduction of inflammatory responses by reducing the adhesiveness of CASMC, a mechanism suggested to be mediated by the production and release of EMAP II.


Women with previous stress fractures show reduced bone material strength.
Background and purpose - Bone fragility is determined by bone mass, bone architecture, and the material properties of bone. Microindentation has been introduced as a measurement method that reflects bone material properties. The pathogenesis of underlying stress fractures, in particular the role of impaired bone material properties, is still poorly understood. Based on the hypothesis that impaired bone material strength might play a role in the development of stress fractures, we used microindentation in patients with stress fractures and in controls. Patients and methods - We measured bone material strength index (BMSi) by microindentation in 30 women with previous stress fractures and in 30 normal controls. Bone mineral density by DXA and levels of the bone markers C-terminal cross-linking telopeptide of type-1 collagen (CTX) and N-terminal propeptide of type-1 procollagen (P1NP) were also determined. Results - Mean BMSi in stress fracture patients was significantly lower than in the controls (SD 72 (8.7) vs. 77 (7.2); p = 0.02). The fracture subjects also had a significantly lower mean bone mineral density (BMD) than the controls (0.9 (0.02) vs. 1.0 (0.06); p = 0.03). Bone turnover-as reflected in serum levels of the bone marker CTX-was similar in both groups, while P1NP levels were significantly higher in the women with stress fractures (55 μg/L vs. 42 μg/L; p = 0.03). There was no correlation between BMSi and BMD or bone turnover. Interpretation - BMSi was inferior in patients with previous stress fracture, but was unrelated to BMD and bone turnover. The lower values of BMSi in patients with previous stress fracture combined with a lower BMD may contribute to the increased propensity to develop stress fractures in these patients.


Fertility rate trends among adolescent girls with major mental illness: a population-based study.
Fertility rates among adolescents have decreased substantially in recent years, yet fertility rates among adolescent girls with mental illness have not been studied. We examined temporal trends in fertility rates among adolescent girls with major mental illness. We conducted a repeated annual cross-sectional study of fertility rates among girls aged 15 to 19 years in Ontario, Canada (1999-2009). Girls with major mental illness were identified through administrative health data indicating the presence of a psychotic, bipolar, or major depressive disorder within 5 years preceding pregnancy (60,228 person-years). The remaining girls were classified into the comparison group (4,496,317 person-years). The age-specific fertility rate (number of live births per 1000 girls) was calculated annually and by using 3-year moving averages for both groups. The incidence of births to girls with major mental illness was 1 in 25. The age-specific fertility rate for girls with major mental illness was 44.9 per 1000 (95% confidence interval [CI]: 43.3-46.7) compared with 15.2 per 1000 (95% CI: 15.1-15.3) in unaffected girls (rate ratio: 2.95; 95% CI: 2.84-3.07). Over time, girls with major mental illness had a smaller reduction in fertility rate (relative rate: 0.86; 95% CI: 0.78-0.96) than did unaffected girls (relative rate: 0.78; 95% CI: 0.76-0.79). These results have key clinical and public policy implications. Our findings highlight the importance of considering major mental illness in the design and implementation of pregnancy prevention programs as well as in targeted antenatal and postnatal programs to ensure maternal and child well-being.


Fruit and Vegetable Intake and Mortality in Adults undergoing Maintenance Hemodialysis.
Higher fruit and vegetable intake is associated with lower cardiovascular and all-cause mortality in the general population. It is unclear whether this association occurs in patients on hemodialysis, in whom high fruit and vegetable intake is generally discouraged because of a potential risk of hyperkalemia. We aimed to evaluate the association between fruit and vegetable intake and mortality in hemodialysis. Fruit and vegetable intake was ascertained by the Global Allergy and Asthma European Network food frequency questionnaire within the Dietary Intake, Death and Hospitalization in Adults with ESKD Treated with Hemodialysis study, a multinational cohort study of 9757 adults on hemodialysis, of whom 8078 (83%) had analyzable dietary data. Adjusted Cox regression analyses clustered by country were conducted to evaluate the association between tertiles of fruit and vegetable intake with all-cause, cardiovascular, and noncardiovascular mortality. Estimates were calculated as hazard ratios with 95% confidence intervals (95% CIs). During a median follow up of 2.7 years (18,586 person-years), there were 2082 deaths (954 cardiovascular). The median (interquartile range) number of servings of fruit and vegetables was 8 (4-14) per week; only 4% of the study population consumed at least four servings per day as recommended in the general population. Compared with the lowest tertile of servings per week (0-5.5, median 2), the adjusted hazard ratios for the middle (5.6-10, median 8) and highest (>10, median 17) tertiles were 0.90 (95% CI, 0.81 to 1.00) and 0.80 (95% CI, 0.71 to 0.91) for all-cause mortality, 0.88 (95% CI, 0.76 to 1.02) and 0.77 (95% CI, 0.66 to 0.91) for noncardiovascular mortality and 0.95 (95% CI, 0.81 to 1.11) and 0.84 (95% CI, 0.70 to 1.00) for cardiovascular mortality, respectively. Fruit and vegetable intake in the hemodialysis population is low and a higher consumption is associated with lower all-cause and noncardiovascular death.


The incidence of type-1 diabetes in NOD mice is modulated by restricted flora not germ-free conditions.
In the NOD mouse, the incidence of type-1 diabetes is thought to be influenced by the degree of cleanliness of the mouse colony. Studies collectively demonstrate that exposure to bacterial antigen or infection in the neonatal period prevents diabetes [1], [2], [3], [4], [5], [6], [7], [8], [9], [10], supporting the notion that immunostimulation can benefit the maturation of the postnatal immune system [11]. A widely accepted extrapolation from this data has been the notion that NOD mice maintained under germ-free conditions have an increased incidence of diabetes. However, evidence supporting this influential concept is surprisingly limited [12]. In this study, we demonstrate that the incidence of diabetes in female NOD mice remained unchanged under germ-free conditions. By contrast, a spontaneous monoculture with a gram-positive aerobic spore-forming rod delayed the onset and reduced the incidence of diabetes. These findings challenge the view that germ-free NOD mice have increased diabetes incidence and demonstrate that modulation of intestinal microbiota can prevent the development of type-1 diabetes.


Inhibition of human UDP-glucuronosyltransferase enzymes by lapatinib, pazopanib, regorafenib and sorafenib: Implications for hyperbilirubinemia.
Kinase inhibitors (KIs) are a rapidly expanding class of drugs used primarily for the treatment of cancer. Data relating to the inhibition of UDP-glucuronosyltransferase (UGT) enzymes by KIs is sparse. However, lapatinib (LAP), pazopanib (PAZ), regorafenib (REG) and sorafenib (SOR) have been implicated in the development of hyperbilirubinemia in patients. This study aimed to characterise the role of UGT1A1 inhibition in hyperbilirubinemia and assess the broader potential of these drugs to perpetrate drug-drug interactions arising from UGT enzyme inhibition. Twelve recombinant human UGTs from subfamilies 1A and 2B were screened for inhibition by LAP, PAZ, REG and SOR. IC50 values for the inhibition of all UGT1A enzymes, except UGT1A3 and UGT1A4, by the four KIs were <10μM. LAP, PAZ, REG and SOR inhibited UGT1A1-catalysed bilirubin glucuronidation with mean IC50 values ranging from 34nM (REG) to 3734nM (PAZ). Subsequent kinetic experiments confirmed that REG and SOR were very potent inhibitors of human liver microsomal β-estradiol glucuronidation, an established surrogate for bilirubin glucuronidation, with mean Ki values of 20 and 33nM, respectively. Ki values for LAP and PAZ were approximately 1- and 2-orders of magnitude higher than those for REG and SOR. REG and SOR were equipotent inhibitors of human liver microsomal UGT1A9 (mean Ki 678nM). REG and SOR are the most potent inhibitors of a human UGT enzyme identified to date. In vitro-in vivo extrapolation indicates that inhibition of UGT1A1 contributes significantly to the hyperbilirubinemia observed in patients treated with REG and SOR, but not with LAP and PAZ. Inhibition of other UGT1A1 substrates in vivo is likely.


Kinetics of intraperitoneally infused insulin in rats. Functional implications for the bioartificial pancreas.
Intraperitoneal transplantation of encapsulated islets can restore normoglycemia in diabetic recipients but not normal glucose tolerance nor normal insulin responses to a physiological stimulus. This study investigates whether the intraperitoneal implantation site as such contributes to the interference with optimal transport kinetics between the islets and the bloodstream. Insulin was infused into the peritoneal cavity of conscious and freely moving rats in doses of 20, 40, and 80 pmol.l-1.min-1 during 15 min, to mimic the gradual release of insulin from an encapsulated, i.e., a nonvascularized, islet graft. With 20 pmol.l-1.min-1, we observed virtually no rise of insulin levels, and it took 30 min until glucose levels had dropped significantly. With 40 and 80 pmol.l-1.min-1 insulin infusions, there was a dose-dependent rise of insulin and decrease of glucose levels. When compared with intraportal infusions with the same insulin dosages, however, they were strongly delayed and reduced as well as prolonged. Similar results were obtained when inulin instead of insulin was intraperitoneally infused, with indicates that the transport of insulin from the peritoneal cavity to the bloodstream is mainly by passive diffusion. With a view on the clinical efficacy of the bioartificial pancreas, our findings indicate that we should focus on finding or creating a transplantation site that, more than the unmodified peritoneal cavity, permits close contact between the bloodstream and the encapsulated islet tissue.


P-REX2a driving tumorigenesis by PTEN inhibition.
The phosphatase PTEN (phosphatase and tensin homolog deleted on chromosome 10) antagonizes phosphoinositide 3-kinase (PI3K) signaling and is one of the most frequently mutated tumor suppressors in human cancers. Its regulation appears complex and is of great potential clinical importance. The protein P-REX2a (phosphatidylinositol 3,4,5-trisphosphate Rac exchanger 2a), better known as a regulator of the small guanosine triphosphatase Rac, has been identified as a direct regulator of PTEN activity and as a potential oncoprotein. P-REX2a can stimulate cell proliferation by inhibiting PTEN and stimulating downstream PI3K-dependent signaling. This suggests that aberrant control of PTEN by P-REX2a may represent a key tumorigenic mechanism, in agreement with recent studies supporting the pathological relevance of several other proposed PTEN regulators.


Two-aperture method of producing light beams with odd-symmetry for alignment and straightness measurements.
A light beam generated by two laser-illuminated circular apertures in a plane screen and two properly adjusted phase-shifting glass plates exhibits a central irradiance minimum that makes it favorable for alignment applications. The far-field transverse irradiance distribution in such a beam is here determined theoretically and experimentally to facilitate appraisal of its suitability for this purpose. Criteria aiding optimization of the beam and factors that degrade the beam and produce alignment errors are discussed. Alignment accuracy depends upon correspondence of a reference beam to a mathematically ideal, perfectly symmetrical shape. It is shown that a beam closely approaching the ideal can be easily produced by this method. Alignment accuracy is then found to be limited chiefly by air turbulence and stray light, which in the experimental investigation limited precision to +/- 10 /microm at 8.2 m.


No cumulative effect of 10 years of elevated [CO2 ] on perennial plant biomass components in the Mojave Desert.
Elevated atmospheric CO2 concentrations ([CO2 ]) generally increase primary production of terrestrial ecosystems. Production responses to elevated [CO2 ] may be particularly large in deserts, but information on their long-term response is unknown. We evaluated the cumulative effects of elevated [CO2 ] on primary production at the Nevada Desert FACE (free-air carbon dioxide enrichment) Facility. Aboveground and belowground perennial plant biomass was harvested in an intact Mojave Desert ecosystem at the end of a 10-year elevated [CO2 ] experiment. We measured community standing biomass, biomass allocation, canopy cover, leaf area index (LAI), carbon and nitrogen content, and isotopic composition of plant tissues for five to eight dominant species. We provide the first long-term results of elevated [CO2 ] on biomass components of a desert ecosystem and offer information on understudied Mojave Desert species. In contrast to initial expectations, 10 years of elevated [CO2 ] had no significant effect on standing biomass, biomass allocation, canopy cover, and C : N ratios of above- and belowground components. However, elevated [CO2 ] increased short-term responses, including leaf water-use efficiency (WUE) as measured by carbon isotope discrimination and increased plot-level LAI. Standing biomass, biomass allocation, canopy cover, and C : N ratios of above- and belowground pools significantly differed among dominant species, but responses to elevated [CO2 ] did not vary among species, photosynthetic pathway (C3 vs. C4 ), or growth form (drought-deciduous shrub vs. evergreen shrub vs. grass). Thus, even though previous and current results occasionally show increased leaf-level photosynthetic rates, WUE, LAI, and plant growth under elevated [CO2 ] during the 10-year experiment, most responses were in wet years and did not lead to sustained increases in community biomass. We presume that the lack of sustained biomass responses to elevated [CO2 ] is explained by inter-annual differences in water availability. Therefore, the high frequency of low precipitation years may constrain cumulative biomass responses to elevated [CO2 ] in desert environments.


What are the important manoeuvres for beginners to minimize surgical time in primary total knee arthroplasty?
It was hypothesized that surgical time of beginners would be much longer than that of experts. Our purpose was to investigate and clarify the important manoeuvres for beginners to minimize surgical time in primary total knee arthroplasty (TKA) as a multicentre study. A total of 300 knees in 248 patients (averaged 74.6 years) were enrolled. All TKAs were done using the same instruments and the same measured resection technique at 14 facilities by 25 orthopaedic surgeons. Surgeons were divided into three surgeon groups (four experts, nine medium-volume surgeons and 12 beginners). The surgical technique was divided into five phases. Detailed surgical time and ratio of the time in each phase to overall surgical time were recorded and compared among the groups in each phase. A total of 62, 119, and 119 TKAs were done by beginners, medium-volume surgeons, and experts, respectively. Significant differences in surgical time among the groups were seen in each phase. Concerning the ratio of the time, experts and medium-volume surgeons seemed cautious in fixation of the permanent component compared to other phases. Interestingly, even in ratio, beginners and medium-volume surgeons took more time in exposure of soft tissue compared to experts. (0.14 in beginners, 0.13 in medium-volume surgeons, 0.11 in experts, P < 0.05). Beginners took more time in exposure and closure of soft tissue compared to experts. Improvement in basic technique is essential to minimize surgical time among beginners. First of all, surgical instructors should teach basic techniques in primary TKA for beginners. Therapeutic studies, Level IV.


Apixaban versus enoxaparin for thromboprophylaxis after hip replacement.
There are various regimens for thromboprophylaxis after hip replacement. Low-molecular-weight heparins such as enoxaparin predominantly inhibit factor Xa but also inhibit thrombin to some degree. Orally active, specific factor Xa inhibitors such as apixaban may provide effective thromboprophylaxis with a lower risk of bleeding and improved ease of use. In this double-blind, double-dummy study, we randomly assigned 5407 patients undergoing total hip replacement to receive apixaban at a dose of 2.5 mg orally twice daily or enoxaparin at a dose of 40 mg subcutaneously every 24 hours. Apixaban therapy was initiated 12 to 24 hours after closure of the surgical wound; enoxaparin therapy was initiated 12 hours before surgery. Prophylaxis was continued for 35 days after surgery, followed by bilateral venographic studies. The primary efficacy outcome was the composite of asymptomatic or symptomatic deep-vein thrombosis, nonfatal pulmonary embolism, or death from any cause during the treatment period. Patients were followed for an additional 60 days after the last intended dose of study medication. A total of 1949 patients in the apixaban group (72.0%) and 1917 patients in the enoxaparin group (71.0%) could be evaluated for the primary efficacy analysis. The primary efficacy outcome occurred in 27 patients in the apixaban group (1.4%) and in 74 patients in the enoxaparin group (3.9%) (relative risk with apixaban, 0.36; 95% confidence interval [CI], 0.22 to 0.54; P<0.001 for both noninferiority and superiority; absolute risk reduction, 2.5 percentage points; 95% CI, 1.5 to 3.5). The composite outcome of major and clinically relevant nonmajor bleeding occurred in 129 of 2673 patients assigned to apixaban (4.8%) and 134 of 2659 assigned to enoxaparin (5.0%) (absolute difference in risk, -0.2 percentage points; 95% CI, -1.4 to 1.0). Among patients undergoing hip replacement, thromboprophylaxis with apixaban, as compared with enoxaparin, was associated with lower rates of venous thromboembolism, without increased bleeding. (Funded by Bristol-Myers Squibb and Pfizer; ClinicalTrials.gov number, NCT00423319.).


Investigation of Japanese encephalitis virus as a cause of acute encephalitis in southern Pakistan, April 2015-January 2018.
Japanese encephalitis (JE) occurs in fewer than 1% of JE virus (JEV) infections, often with catastrophic sequelae including death and neuropsychiatric disability. JEV transmission in Pakistan was documented in 1980s and 1990s, but recent evidence is lacking. Our objective was to investigate JEV as a cause of acute encephalitis in Pakistan. Persons aged ≥1 month with possible JE admitted to two acute care hospitals in Karachi, Pakistan from April 2015 to January 2018 were enrolled. Cerebrospinal fluid (CSF) or serum samples were tested for JEV immunoglobulin M (IgM) using the InBios JE DetectTM assay. Positive or equivocal samples had confirmatory testing using plaque reduction neutralization tests. Among 227 patients, testing was performed on CSF in 174 (77%) and on serum in 53 (23%) patients. Six of eight patient samples positive or equivocal for JEV IgM had sufficient volume for confirmatory testing. One patient had evidence of recent West Nile virus (WNV) neurologic infection based on CSF testing. One patient each had recent dengue virus (DENV) infection and WNV infection based on serum results. Recent flavivirus infections were identified in two persons, one each based on CSF and serum results. Specific flaviviruses could not be identified due to serologic cross-reactivity. For the sixth person, JEV neutralizing antibodies were confirmed in CSF but there was insufficient volume for further testing. Hospital-based JE surveillance in Karachi, Pakistan could not confirm or exclude local JEV transmission. Nonetheless, Pakistan remains at risk for JE due to presence of the mosquito vector, amplifying hosts, and rice irrigation. Laboratory surveillance for JE should continue among persons with acute encephalitis. However, in view of serological cross-reactivity, confirmatory testing of JE IgM positive samples at a reference laboratory is essential.


Clinical risk modification, quality, and patient safety: interrelationships, problems, and future potential.
Iatrogenic injury, in which patients are unintentionally injured by medical treatment, occurs in 4% of hospital admissions and causes considerable human suffering, financial losses, and waste of healthcare resources. This article discusses why existing quality initiatives have had little impact on iatrogenic injury and suggests an approach to clinical risk modification that may enhance the safety of medical treatment.


Variance component analysis of allozyme frequency data from eastern populations of Drosophila melanogaster.
Gene frequency variation at eight polymorphic allozyme loci in Drosophila melanogaster populations in North Carollina and the east coast of the United States were analyzed utilizing the variance component estimation procedures suggested by Cockerham (1969, 1973). These variance components were used to estimate correlations of genes within small geographic regions. The average (over loci) correlation between genes in the same individual within subpopulations was estimated to be 0.033. That between genes in the same subpopulation in different individuals was estimated to be very small, although significantly different from zero. The macrogeographic variation measured by the correlation of genes sampled from the same local region was large for some loci and smaller for others. this variation was also analyzed by correlation with latitude and longitude. Several previously recognized clines were identified as were several new clines.--These results were interpreted as indicating either some degree of nonrandom mating and local breeding unit isolation or a low frequency of null alleles. The geographic and temporal variation has no simple interpretation.


Cognitive functioning in Behçet's disease.
The impact of Behçet's disease on higher cognitive functions is still poorly understood. We proposed (1) to characterize the neuropsychological profile of Behçet's disease patients with (Neuro-BD) and without (BD) neurological manifestations; (2) to identify which clinical, psychopathological, and genetic variables are related to neuropsychological performance; and (3) to explore the association between cognitive functioning and neuroimaging findings in BD patients. Fifteen Neuro-BD and 35 BD patients in the nonactive phase of their illness underwent a neurological examination, performed a comprehensive battery of neuropsychological tests, and answered the hospital anxiety and depression scale. Human leukocyte antigen (HLA)-B*51 genotyping was also performed. Patients' neuropsychological performances were compared to those of healthy demographically matched subjects. Within one month from the testing date, a subset of 20-BD patients underwent a magnetic resonance imaging (MRI) scan. Fifty-three percent of Neuro-BD and 40% of BD patients were impaired at least on one neuropsychological measure (i.e., digit span-forward). Poorer cognitive functioning in Neuro-BD was associated with parenchymal involvement, whereas in BD it was related to presence of white matter changes in the frontal lobes, history of headache complaints, or higher levels of anxiety and depression. Current prednisone intake had a positive impact on neuropsychological performance. Disease duration, time since onset of neurological manifestations, or presence of HLA-B*51 allele had no significant influence. Our results indicate that Behçet's disease may affect cognitive abilities in the absence of overt neurological symptoms. These findings point to an insidious course of neurological involvement.


Comparative Chromosome Painting in Genets (Carnivora, Viverridae, Genetta), the Only Known Feliforms with a Highly Rearranged Karyotype.
Mammalian carnivores have been extensively studied by cross-species chromosome painting, which indicated a high degree of karyotypic conservatism in the cat-like suborder Feliformia relative to the ancestral carnivore karyotype (ACK). The first exception to this high degree of karyotypic conservation in feliforms was recently confirmed in genets, mesocarnivores belonging to the basal family Viverridae. Here, we present a comparative analysis of the chromosome rearrangements among 2 subspecies of the small-spotted genet Genetta genetta (the Iberian nominate and the Arabian grantii) and the panther genet G. maculata, the 2 most common and widespread genets, using whole-chromosome paints from the domestic cat (Felis catus). The chromosome homology maps and the presence of numerous interstitial telomeric sites in both genet species strengthen the hypothesis that a highly rearranged karyotype compared to the ACK may occur throughout Genetta. The karyotype of G. maculata appears to have undergone more rearrangements than that of G. genetta, which is an older lineage. Notably, we identified a tandem fusion distinguishing G. g. genetta and G. g.grantii. As G. g. grantii is morphologically and genetically distinctive, and tandem fusions have been associated with substantial postzygotic isolation in mammals, this cytogenetic finding flags the subspecies for future taxonomic investigations.


Photophysical studies of dipolar organic dyes that feature a 1,3-cyclohexadiene conjugated linkage: the implication of a twisted intramolecular charge-transfer state on the efficiency of dye-sensitized solar cells.
A detailed study of the synthesis and photophysical properties of a new series of dipolar organic photosensitizers that feature a 1,3-cyclohexadiene moiety integrated into the π-conjugated structural backbone has been carried out. Dye-sensitized solar cells (DSSCs) based on these structurally simple dyes have shown appreciable photo-to-electrical energy conversion efficiency, with the highest one up to 4.03 %. Solvent-dependent fluorescence studies along with the observation of dual emission on dye 4 b and single emission on dyes 4 a and 32 suggest that dye 4 b possesses a highly polar emissive excited state located at a lower-energy position than at the normal emissive excited state. A detailed photophysical investigation in conjunction with computational studies confirmed the twisted intramolecular charge-transfer (TICT) state to be the lowest emissive excited state for dye 4 b in polar solvents. The relaxation from higher-charge-injection excited states to the lowest TICT state renders the back-electron transfer process a forbidden one and significantly retards the charge recombination to boost the photocurrent. The electrochemical impedance under illumination and transient photovoltage decay studies showed smaller charge resistance and longer electron lifetime in 4 b-based DSSC compared to the DSSCs with reference dyes 4 a and 32, which further illustrates the positive influence of the TICT state on the performance of DSSCs.


Computational models to explore the complexity of the epithelial to mesenchymal transition in cancer.
Epithelial to mesenchymal transition (EMT) is a complex biological process that plays a key role in cancer progression and metastasis formation. Its activation results in epithelial cells losing adhesion and polarity and becoming capable of migrating from their site of origin. At this step the disease is generally considered incurable. As EMT execution involves several individual molecular components, connected by nontrivial relations, in vitro techniques are often inadequate to capture its complexity. Computational models can be used to complement experiments and provide additional knowledge difficult to build up in a wetlab. Indeed in silico analysis gives the user total control on the system, allowing to identify the contribution of each independent element. In the following, two kinds of approaches to the computational study of EMT will be presented. The first relies on signal transduction networks description and details how changes in gene expression could influence this process, both focusing on specific aspects of the EMT and providing a general frame for this phenomenon easily comparable with experimental data. The second integrates single cell and population level descriptions in a multiscale model that can be considered a more accurate representation of the EMT. The advantages and disadvantages of each approach will be highlighted, together with the importance of coupling computational and experimental results. Finally, the main challenges that need to be addressed to improve our knowledge of the role of EMT in the neoplastic disease and the scientific and translational value of computational models in this respect will be presented. This article is categorized under: Analytical and Computational Methods > Computational Methods.


Lipid droplet size directs lipolysis and lipophagy catabolism in hepatocytes.
Lipid droplet (LD) catabolism in hepatocytes is mediated by a combination of lipolysis and a selective autophagic mechanism called lipophagy, but the relative contributions of these seemingly distinct pathways remain unclear. We find that inhibition of lipolysis, lipophagy, or both resulted in similar overall LD content but dramatic differences in LD morphology. Inhibition of the lipolysis enzyme adipose triglyceride lipase (ATGL) resulted in large cytoplasmic LDs, whereas lysosomal inhibition caused the accumulation of numerous small LDs within the cytoplasm and degradative acidic vesicles. Combined inhibition of ATGL and LAL resulted in large LDs, suggesting that lipolysis targets these LDs upstream of lipophagy. Consistent with this, ATGL was enriched in larger-sized LDs, whereas lipophagic vesicles were restricted to small LDs as revealed by immunofluorescence, electron microscopy, and Western blot of size-separated LDs. These findings provide new evidence indicating a synergistic relationship whereby lipolysis targets larger-sized LDs to produce both size-reduced and nascently synthesized small LDs that are amenable for lipophagic internalization.


Predicted functions and linkage specificities of the products of the Streptococcus pneumoniae capsular biosynthetic loci.
The sequences of the capsular biosynthetic (cps) loci of 90 serotypes of Streptococcus pneumoniae have recently been determined. Bioinformatic procedures were used to predict the general functions of 1,973 of the 1,999 gene products and to identify proteins within the same homology group, Pfam family, and CAZy glycosyltransferase family. Correlating cps gene content with the 54 known capsular polysaccharide (CPS) structures provided tentative assignments of the specific functions of the different homology groups of each functional class (regulatory proteins, enzymes for synthesis of CPS constituents, polymerases, flippases, initial sugar transferases, glycosyltransferases [GTs], phosphotransferases, acetyltransferases, and pyruvyltransferases). Assignment of the glycosidic linkages catalyzed by the 342 GTs (92 homology groups) is problematic, but tentative assignments could be made by using this large set of cps loci and CPS structures to correlate the presence of particular GTs with specific glycosidic linkages, by correlating inverting or retaining linkages in CPS repeat units with the inverting or retaining mechanisms of the GTs predicted from their CAZy family membership, and by comparing the CPS structures of serotypes that have very similar cps gene contents. These large-scale comparisons between structure and gene content assigned the linkages catalyzed by 72% of the GTs, and all linkages were assigned in 32 of the serotypes with known repeat unit structures. Clear examples where very similar initial sugar transferases or glycosyltransferases catalyze different linkages in different serotypes were also identified. These assignments should provide a stimulus for biochemical studies to evaluate the reactions that are proposed.


Excision of the maize transposable element Ac in flax callus.
The frequency and fidelity of Ac transposition, and that of its non-autonomous derivative Ds, were investigated in flax callus. Flax (Linum usitatissimum var. Antares) hypocotyls were transformed with Agrobacterium Ti plasmid vectors containing the Ac or Ds element inserted within the untranslated leader sequence of a chimaeric neomycin phosphotransferase II gene. Kanamycin resistant tissues were produced as a result of excision of Ac in around 35% of the total number of Ac-containing transformants. In contrast, no excision was observed from transformants containing the Ds element. Whilst Ac appears to have excised completely from T-DNAs, little evidence was found to infer reintegration of the Ac element into the genome.


A nonlinear control method based on ANFIS and multiple models for a class of SISO nonlinear systems and its application.
This paper presents a novel nonlinear control strategy for a class of uncertain single-input and single-output discrete-time nonlinear systems with unstable zero-dynamics. The proposed method combines adaptive-network-based fuzzy inference system (ANFIS) with multiple models, where a linear robust controller, an ANFIS-based nonlinear controller and a switching mechanism are integrated using multiple models technique. It has been shown that the linear controller can ensure the boundedness of the input and output signals and the nonlinear controller can improve the dynamic performance of the closed loop system. Moreover, it has also been shown that the use of the switching mechanism can simultaneously guarantee the closed loop stability and improve its performance. As a result, the controller has the following three outstanding features compared with existing control strategies. First, this method relaxes the assumption of commonly-used uniform boundedness on the unmodeled dynamics and thus enhances its applicability. Second, since ANFIS is used to estimate and compensate the effect caused by the unmodeled dynamics, the convergence rate of neural network learning has been increased. Third, a "one-to-one mapping" technique is adapted to guarantee the universal approximation property of ANFIS. The proposed controller is applied to a numerical example and a pulverizing process of an alumina sintering system, respectively, where its effectiveness has been justified.


Clinical and economic burden of antimicrobial resistance.
Knowledge of the clinical and economic impact of antimicrobial resistance is useful to influence programs and behavior in healthcare facilities, to guide policy makers and funding agencies, to define the prognosis of individual patients and to stimulate interest in developing new antimicrobial agents and therapies. There are a variety of important issues that must be considered when designing or interpreting studies into the clinical and economic outcomes associated with antimicrobial resistance. One of the most misunderstood issues is how to measure cost appropriately. Although imperfect, existing data show that there is an association between antimicrobial resistance in Staphylococcus aureus, enterococci and Gram-negative bacilli and increases in mortality, morbidity, length of hospitalization and cost of healthcare. Patients with infections due to antimicrobial-resistant organisms have higher costs (US $6,000-30,000) than do patients with infections due to antimicrobial-susceptible organisms; the difference in cost is even greater when patients infected with antimicrobial-resistant organisms are compared with patients without infection. Given limited budgets, knowledge of the clinical and economic impact of antibiotic-resistant bacterial infections, coupled with the benefits of specific interventions targeted to reduce these infections, will allow for optimal control and improved patient safety. In this review, the authors discuss a variety of important issues that must be considered when designing or interpreting studies of the clinical and economic outcomes associated with antimicrobial resistance. Representative literature is reviewed regarding the associations between antimicrobial resistance in specific pathogens and adverse outcomes, including increased mortality, length of hospital stay and cost.


A three-dimensional model of human organic anion transporter 1: aromatic amino acids required for substrate transport.
Organic anion transporters (OATs) play a critical role in the handling of endogenous and exogenous organic anions by excretory and barrier tissues. Little is known about the OAT three-dimensional structure or substrate/protein interactions involved in transport. In this investigation, a theoretical three-dimensional model was generated for human OAT1 (hOAT1) based on fold recognition to the crystal structure of the glycerol 3-phosphate transporter (GlpT) from Escherichia coli. GlpT and hOAT1 share several sequence motifs as major facilitator superfamily members. The structural hOAT1 model shows that helices 5, 7, 8, 10, and 11 surround an electronegative putative active site ( approximately 830A(3)). The site opens to the cytoplasm and is surrounded by three residues not previously examined for function (Tyr(230) (domain 5) and Lys(431) and Phe(438) (domain 10)). Effects of these residues on p-aminohippurate (PAH) and cidofovir transport were assessed by point mutations in a Xenopus oocyte expression system. Membrane protein expression was severely limited for the Y230A mutant. For the K431A and F438A mutants, [(3)H]PAH uptake was less than 30% of wild-type hOAT1 uptake after protein expression correction. Reduced V(max) values for the F438A mutant confirmed lower protein expression. In addition, the F438A mutant exhibited an increased affinity for cidofovir but was not significantly different for PAH. Differences in handling of PAH and cidofovir were also observed for the Y230F mutant. Little uptake was determined for cidofovir, whereas PAH uptake was similar to wild-type hOAT1. Therefore, the hOAT1 structural model has identified two new residues, Tyr(230) and Phe(438), which are important for substrate/protein interactions.


Hip and spine bone mineral density are greater in master sprinters, but not endurance runners compared with non-athletic controls.
We examined bone density in older athletes and controls. Sprinters had greater hip and spine bone density than endurance athletes and controls, whereas values were similar in the latter two groups. These results could not be explained by differences in impact, muscle size or power between sprint and endurance athletes. We examined the relationship between prolonged participation in regular sprint or endurance running and skeletal health at key clinical sites in older age, and the factors responsible for any associations which we observed. We recruited 38 master sprint runners (28 males, 10 females, mean age 71 ± 7 years), 149 master endurance runners (111 males, 38 females, mean age 70 ± 6 years) and 59 non-athletic controls (29 males, 30 females, mean age 74 ± 5 years). Dual X-ray absorptiometry was used to assess hip and spine bone mineral density (BMD), body composition (lean and fat mass), whilst jump power was assessed with jumping mechanography. In athletes, vertical impacts were recorded over 7 days from a waist-worn accelerometer, and details of starting age, age-graded performance and training hours were recorded. In ANOVA models adjusted for sex, age, height, body composition, and jump power, sprinter hip BMD was 10 and 14% greater than that of endurance runners and controls respectively. Sprinter spine BMD was also greater than that of both endurance runners and controls. There were no differences in hip or spine BMD between endurance runners and controls. Stepwise regression showed only discipline (sprint/endurance), sex, and age as predictors of athlete spine BMD, whilst these variables and starting age were predictive of hip BMD. Regular running is associated with greater BMD at the fracture-prone hip and spine sites in master sprinters but not endurance runners. These benefits cannot be explained by indicators of mechanical loading measured in this study including vertical impacts, body composition or muscular output.


The impact of Recovery Colleges on mental health staff, services and society.
Recovery Colleges are opening internationally. The evaluation focus has been on outcomes for Recovery College students who use mental health services. However, benefits may also arise for: staff who attend or co-deliver courses; the mental health and social care service hosting the Recovery College; and wider society. A theory-based change model characterising how Recovery Colleges impact at these higher levels is needed for formal evaluation of their impact, and to inform future Recovery College development. The aim of this study was to develop a stratified theory identifying candidate mechanisms of action and outcomes (impact) for Recovery Colleges at staff, services and societal levels. Inductive thematic analysis of 44 publications identified in a systematised review was supplemented by collaborative analysis involving a lived experience advisory panel to develop a preliminary theoretical framework. This was refined through semi-structured interviews with 33 Recovery College stakeholders (service user students, peer/non-peer trainers, managers, community partners, clinicians) in three sites in England. Candidate mechanisms of action and outcomes were identified at staff, services and societal levels. At the staff level, experiencing new relationships may change attitudes and associated professional practice. Identified outcomes for staff included: experiencing and valuing co-production; changed perceptions of service users; and increased passion and job motivation. At the services level, Recovery Colleges often develop somewhat separately from their host system, reducing the reach of the college into the host organisation but allowing development of an alternative culture giving experiential learning opportunities to staff around co-production and the role of a peer workforce. At the societal level, partnering with community-based agencies gave other members of the public opportunities for learning alongside people with mental health problems and enabled community agencies to work with people they might not have otherwise. Recovery Colleges also gave opportunities to beneficially impact on community attitudes. This study is the first to characterise the mechanisms of action and impact of Recovery Colleges on mental health staff, mental health and social care services, and wider society. The findings suggest that a certain distance is needed in the relationship between the Recovery College and its host organisation if a genuine cultural alternative is to be created. Different strategies are needed depending on what level of impact is intended, and this study can inform decision-making about mechanisms to prioritise. Future research into Recovery Colleges should include contextual evaluation of these higher level impacts, and investigate effectiveness and harms.


Static blood flow autoregulation in the optic nerve head in normal and experimental glaucoma.
To characterize the static blood flow autoregulation in the optic nerve head (ONH), and to investigate its role in hemodynamic changes in experimental glaucoma (EG). Unilateral elevation of intraocular pressure (IOP) was induced in 15 adult rhesus macaques by laser treatment to the trabecular meshwork. Prior to and after laser treatment, retinal nerve fiber layer thickness (RNFLT) was assessed, biweekly, by spectral-domain optical coherence tomography. Optic nerve head static autoregulation was assessed by determining the percentage blood flow (BF) change after the IOP was acutely increased from 10 to 30, 40, or 50 mm Hg manometrically, utilizing a laser speckle flowgraphy device. Postlaser IOP (measured during average 7.7 ± 2.6 months) was 20.2 ± 5.9 mm Hg in EG eyes and 12.3 ± 2.6 mm Hg in control eyes (P < 0.0001). Retinal nerve fiber layer thickness was reduced by 33 ± 22% of the baseline values (P < 0.001) on average in EG eyes and by 0.4 ± 2.3% in control eyes (P > 0.05). The ONH BF remained at a constant level within a range of ocular perfusion pressure (OPP), 41 mm Hg and above. The autoregulation curves, created by all 723 tests in control and 352 tests in EG, were not significantly different (P = 0.71). Optic nerve head BF in normal nonhuman primate (NHP) eyes is effectively regulated within a range of OPP approximately 41 mm Hg and above. Chronic IOP elevation causes no remarkable change to the static autoregulation within the ONH of EG eyes.


Asthma and obesity in children: current evidence and potential systems biology approaches.
Both obesity and asthma are highly prevalent, complex diseases modified by multiple factors. Genetic, developmental, lung mechanical, immunological and behavioural factors have all been suggested as playing a causal role between the two entities; however, their complex mechanistic interactions are still poorly understood and evidence of causality in children remains scant. Equally lacking is evidence of effective treatment strategies, despite the fact that imbalances at vulnerable phases in childhood can impact long-term health. This review is targeted at both clinicians frequently faced with the dilemma of how to investigate and treat the obese asthmatic child and researchers interested in the topic. Highlighting the breadth of the spectrum of factors involved, this review collates evidence regarding the investigation and treatment of asthma in obese children, particularly in comparison with current approaches in 'difficult-to-treat' childhood asthma. Finally, the authors propose hypotheses for future research from a systems-based perspective.


A phase III, double-blind, placebo-controlled, multicenter study on the efficacy of recombinant human antithrombin in heparin-resistant patients scheduled to undergo cardiac surgery necessitating cardiopulmonary bypass.
The study evaluated the efficacy of recombinant human antithrombin (rhAT) for restoring heparin responsiveness in heparin resistant patients undergoing cardiac surgery. This was a multicenter, randomized, double-blind, placebo-controlled study in heparin-resistant patients undergoing cardiac surgery with cardiopulmonary bypass. Heparin resistance was diagnosed when the activated clotting time was less than 480 s after 400 U/kg heparin. Fifty-four heparin-resistant patients were randomized. One cohort received 75 U/kg rhAT, and the other received normal saline. If the activated clotting time remained less than 480 s, this was considered treatment failure, and 2 units fresh frozen plasma was transfused. Patients were monitored for adverse events. Only 19% of patients in the rhAT group received fresh frozen plasma, compared with 81% of patients in the placebo group (P < 0.001). During their hospitalization, 48% of patients in the rhAT group received fresh frozen plasma, compared with 85% of patients in the placebo group (P = 0.009). Patients in the placebo group required higher heparin doses (P < 0.005) for anticoagulation. There was no increase in serious adverse events associated with rhAT. There was increased blood loss 12 h postoperatively (P = 0.05) with a trend toward increased 24-h bleeding in the rhAT group (P = 0.06). There was no difference between the groups in blood and platelet transfusions. Treatment with 75 U/kg rhAT is effective in restoring heparin responsiveness and promoting therapeutic anticoagulation in the majority of heparin-resistant patients. Treating heparin-resistant patients with rhAT may decrease the requirement for heparin and fresh frozen plasma.


Fast and Sensitive Detection of Oligosaccharides Using Desalting Paper Spray Mass Spectrometry (DPS-MS).
Conventional mass spectrometry (MS)-based analytical methods for small carbohydrate fragments (oligosaccharides, degree of polymerization 2-12) are time-consuming due to the need for an offline sample pretreatment such as desalting. Herein, we report a new paper spray ionization method, named desalting paper spray (DPS), which employs a piece of triangular filter paper for both sample desalting and ionization. Unlike regular paper spray ionization (PSI) and nanoelectrospray ionization (nanoESI), DPS-MS allows fast and sensitive detection of oligosaccharides in biological samples having complex matrices (e.g., Tris, PBS, HEPES buffers, or urine). When an oligosaccharide sample is loaded onto the filter paper substrate (10 × 5 mm, height × base) made mostly of cellulose, oligosaccharides are adsorbed on the paper via hydrophilic interactions with cellulose. Salts and buffers can be washed away using an ACN/H2O (90/10 v/v) solution, while oligosaccharides can be eluted from the paper using a solution of ACN/H2O/formic acid (FA) (10/90/1 v/v/v) and directly spray-ionized from the tip of the paper. Various saccharides at trace levels (e.g., 50 fmol) in nonvolatile buffer can be quickly analyzed by DPS-MS (<5 min per sample). DPS-MS is also applicable for direct detection of oligosaccharides from glycosyltransferase (GT) reactions, a challenging task that typically requires a radioactive assay. Quantitative analysis of acceptor and product oligosaccharides shows increased product with increased GT enzymes used for the reaction, a result in line with the radioactivity assay. This work suggests that DPS-MS has potential for rapid oligosaccharide analysis from biological samples.


Longitudinal changes in dietary fat intake and associated changes in cardiovascular risk factors in adults with type 2 diabetes: the ACCORD trial.
To measure dietary fat intake using the Puget Sound Eating Patterns (PEP) questionnaire, a validated 19-item food questionnaire, and to quantify how reduced dietary fat intake affects cardiovascular risk factors in adults with type 2 diabetes. Randomized controlled trial including a subsample of 1781 Action to Control Cardiovascular Risk in Diabetes (ACCORD) participants. Participants received dietary counseling to consume a reduced-fat diet. Outcome measures included HbA1c, fasting lipid profile, blood pressure, and weight. Longitudinal linear regression analyses were used to evaluate relationships between baseline and follow-up PEP scores and cardiovascular risk factors. PEP scores decreased significantly from baseline to 12-month follow up with a mean difference of -0.09 ± 0.39, P<0.001. All of the fat intake subscales showed significant improvement at 12 months from baseline. White race, female gender, and more hours per week of physical activity were correlated with a decline in PEP scores at 1-year. A longitudinal decrease in dietary fat intake was associated with significantly less weight gain at 12- and 36-months and lower serum triglycerides at 1 year. Reduced fat intake as measured by a brief questionnaire was associated with significant improvement in some cardiovascular risk factors (triglycerides and weight), but not in others.


Dry needling technique decreases spasticity and improves general functioning in incomplete spinal cord injury: A case report.
Context: Spasticity in neurological disorders (i.e. stroke patients and cerebral palsy) is positively improved by dry needling. However, reports are scarce regarding the potential effects of dry needling in reducing spasticity and improving functionality in patients with an incomplete spinal cord injury. The aim of this case report was to study the immediate, short-term effects of dry needling treatment (10 weeks) on spasticity, dynamic stability, walking velocity, self-independence, and pain in a single patient with an incomplete spinal cord injury.Findings: The dry needling treatment resulted in immediate, short-time effects on basal spasticity in the upper (reduction from 2 to 0 point median) and lower (reduction from 2 to 0 point median) limbs, as measured by the modified Ashworth Scale. Dynamic-stability, assessed by trunk accelerometry, improved more than 50% (Root Mean Squared of acceleration, Root Mean Squared of Jerk and step variability), and gait speed improved by 24.7 s (i.e. time to walk 20 m). Self-independence and pain were respectively scored by the Spinal Cord Independence Measure (21 points improvement) and visual analog scale (4 points improvement).Conclusions: This case report demonstrates that dry needling treatment can have positive effects on spasticity, dynamic stability, walking velocity, self-independence, and pain in patients with incomplete spinal cord injury. Further research is needed in a larger patient population to deeply understand the mechanism(s) associated with the obtained results and regarding the clinical significances of dry needling treatment for incomplete spinal cord injury.


Long-term angiographic results of endovascularly "cured" intracranial dural arteriovenous fistulas.
Dural arteriovenous fistulas (DAVFs) are complex lesions consisting of abnormal connections between meningeal arteries and dural venous sinuses and/or cerebral veins. The goal of treatment is surgical or endovascular occlusion of the fistula or fistulous nidus or at least the disconnection of the feeding vessels and the draining veins. Delayed angiographic data on previously embolized dural fistulas is lacking. The authors report their experience and the long-term angiographic results with embolization of intracranial DAVF using Onyx. All cases of DAVF treated primarily with Onyx at the authors' institution from 2006 to 2013 were retrospectively reviewed. Patient demographics, fistula characteristics, embolization details, and angiographic follow-up were analyzed. Fifty-eight patients with DAVFs were treated during the study period. Twenty-two patients were treated with open surgery with or without prior embolization. Thirty-six patients were treated with embolization alone, of whom 26 underwent an attempt at curative embolization and are the subject of this review. All but 2 of these patients were treated in a single session. Angiographic "cure" was achieved in all cases following treatment. Follow-up angiography was performed in 21 patients at a mean of 14 months after treatment (range 2-39 months). Asymptomatic angiographic recurrence of the fistula was evident in 3 of the 21 patients (14.3%). On reviewing the procedural angiograms of the cases in which the DAVFs recurred, it was observed that the Onyx cast did not reach the venous portion in 1 case, whereas it did reach the vein in the other 2 cases. Recurrence following initial angiographic cure of DAVF is not uncommon. Incomplete penetration of the embolic material into the proximal portion of the venous outlet may lead to delayed recurrence. Long-term angiographic follow-up is highly recommended.


An evaluation of the predictive validity of the SAVRY and YLS/CMI in justice-involved youth with fetal alcohol spectrum disorder.
Despite the high prevalence of fetal alcohol spectrum disorder (FASD) in youth criminal justice settings, there is currently no research supporting the use of violence risk assessment tools in this population. This study examined the predictive validity of the Structured Assessment of Violence Risk in Youth (SAVRY) and the Youth Level of Service/Case Management Inventory (YLS/CMI) in justice-involved youth with FASD. Participants were 100 justice-involved youth (ages 12-23; 81% male), including 50 diagnosed with FASD and 50 without FASD or prenatal alcohol exposure. The SAVRY and YLS/CMI were prospectively coded based on interview and file review, with recidivism (both any and violent specifically) coded 1-year post-baseline assessment. Results provide preliminary support for the validity of the SAVRY and YLS/CMI in predicting recidivism in justice-involved youth with FASD. Higher ratings across SAVRY and YLS/CMI domains were found in youth with FASD, underscoring a critical need for assessments and interventions to buffer recidivism risk and address clinical needs. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Beyond social learning.
Cultural evolution requires the social transmission of information. For this reason, scholars have emphasized social learning when explaining how and why culture evolves. Yet cultural evolution results from many mechanisms operating in concert. Here, we argue that the emphasis on social learning has distracted scholars from appreciating both the full range of mechanisms contributing to cultural evolution and how interactions among those mechanisms and other factors affect the output of cultural evolution. We examine understudied mechanisms and other factors and call for a more inclusive programme of investigation that probes multiple levels of the organization, spanning the neural, cognitive-behavioural and populational levels. To guide our discussion, we focus on factors involved in three core topics of cultural evolution: the emergence of culture, the emergence of cumulative cultural evolution and the design of cultural traits. Studying mechanisms across levels can add explanatory power while revealing gaps and misconceptions in our knowledge. This article is part of the theme issue 'Foundations of cultural evolution'.


Prehospital Transfusion of Low-Titer O + Whole Blood for Severe Maternal Hemorrhage: A Case Report.
Introduction: Beginning in 2017, multiple stakeholders within the Southwest Texas Regional Advisory Council for Trauma collaborated to incorporate cold-stored low-titer O RhD-positive whole blood (LTO + WB) into all phases of their trauma system, including the prehospital phase of care. Although the program was initially focused on trauma resuscitation, it was expanded to included non-traumatic hemorrhagic shock patients that may benefit from whole blood resuscitation.Case Report: We report the case of a patient with severe maternal hemorrhage secondary to placenta accreta who received a prehospital transfusion of LTO + WB. We believe this to be the first reported case of post-partum hemorrhage resuscitated out of hospital with whole blood.Discussion: This case highlights the potential benefits of a prehospital whole blood program as well as the controversy surrounding a LTO + WB program that includes females of childbearing age.


Consumption of Coffee but Not of Other Caffeine-Containing Beverages Reduces the Risk of End-Stage Renal Disease in the Singapore Chinese Health Study.
Cross-sectional studies suggest that coffee drinking is associated with better renal function. However, to our knowledge, no prospective study has examined its relation with the risk of end-stage renal disease (ESRD). We examined the relations between coffee, tea, soda, and total caffeine consumption and the risk of ESRD among middle-aged and older Chinese in Singapore. We used data from the Singapore Chinese Health Study, a prospective cohort of 63,257 men and women aged 45-74 y at recruitment from 1993 to 1998. Baseline information on the consumption of caffeinated coffee and other caffeinated beverages (tea and sodas), habitual diet, medical history, and lifestyle factors was obtained via in-person interviews. The standard serving size of 1 cup was assigned as 237 mL in the questionnaire. Incident ESRD cases were identified via linkage with the nationwide registry. We used multivariable Cox regression models to estimate HRs and 95% CIs of ESRD risk associated with the consumption of caffeinated beverages, with adjustment for potential confounders. After a mean follow-up of 16.8 y, 1143 cohort subjects developed ESRD. Compared with those who drank coffee less than daily, the HR (95% CI) was 0.91 (0.79, 1.05) for those who drank 1 cup of coffee/d and 0.82 (0.71, 0.96) for those who drank ≥2 cups/d (P-trend = 0.012). When stratified by sex, this association was observed in men but not in women. Compared with those who drank less than daily, the HR (95% CI) for drinking ≥2 cups/d was 0.71 (0.57, 0.87) among men and 0.97 (0.78, 1.19) among women (P-interaction = 0.03). Conversely, intakes of tea, soda, or total caffeine were not associated with the risk of ESRD in multivariable models. The consumption of ≥2 cups of coffee/d may reduce the risk of ESRD in the general population, especially among men. This study was registered at http://www.clinicaltrials.gov as NCT03356340.


Biochemical physiology of a respiration-deficient mutant of the photosynthetic bacterium Rhodopseudomonas capsulata.
Nonsulfur purple photosynthetic bacteria generally possess the adaptive capacity to grow anaerobically (photosynthetically) and aerobically in darkness (energy obtained by respiratory phosphorylation). To develop a test system for study of interactions between the photosynthetic energy-conversion and dark respiration systems and the mechanism by which O(2) inhibits bacteriochlorophyll synthesis, we have isolated a mutant of Rhodopseudomonas capsulata that is incapable of dark aerobic growth. Both the parent strain and the mutant strain Z-1 (aer-13) grow readily anaerobically in the light with either malate or succinate as the source of carbon and reducing power. They differ, however, in that membrane fragments ("chromatophores") from the mutant show severely impaired ability to use reduced nicotinamideadenine dinucleotide (NADH) as a source of electrons for respiration, oxidative phosphorylation, or "redox buffering" of the photophosphorylation system. The results presented indicate that the genetic lesion responsible for greatly diminished capacity to oxidize NADH can account for inability of Rps. capsulata Z-1 (aer-13) to grow aerobically in darkness.


Delhi's health system exceptionalism: inadequate progress for a global capital city.
India has proclaimed commitment to the goal of Universal Health Coverage and Delhi, the National Capital Territory, has increased investment in public health and other health services over the past decade. The research investigates whether Delhi's increased investment in health over this period is associated with a reduction in premature deaths, after the age of 1 year, which could have been avoided with better access to effective health care interventions (amenable mortality). A population-based study of changes in amenable mortality (AM) in Delhi over the 2003-2013 period. To calculate AM, a list of International Classification of Disease (ICD) codes from the published literature was relied upon. In defining AM in India, an upper age limit of 69 years was adopted, rather than the more common limit of 74 years. Population estimates and vital statistics were downloaded from the Delhi Statistical Handbook. Deaths by cause and age, including medical certification, are from the Vital Statistics site of the Delhi Government. To age-adjust these data, the direct method was employed, using weights derived from the 2010 United Nations world standard population. The research found that, between 2004 and 2013, the age-adjusted rate of AM rose from 0.87 to 1.09. The leading causes of death in both years were septicemia and tuberculosis. Maternal mortality is well above the global level for middle-income countries. Recent investments in public health and health care and the capacity to leverage them to improve access to effective care have not been sufficient to overcome the crushing poverty and inequalities within Delhi. Large and growing numbers of residents die prematurely each year due to causes that are amenable to public health and health care interventions.


Calcium dependence of serotonin-induced changes in rabbit ileal electrolyte transport.
These studies describe the calcium dependence of the serotonin-induced changes in active electrolyte transport in rabbit ileum in vitro. In the presence of a standard calcium concentration (1.2 mM) in the serosal bathing fluid, serosal serotonin caused a transient increase in short-circuit current and a prolonged decrease in net Na and Cl fluxes. Removing calcium from the serosal (no calcium plus 1 mM EGTA) but not the mucosal bathing fluid inhibited the serotoin-induced increase in ileal short-circuit current, and also completely blocked the serotonin effects on net Na and net Cl fluxes. This inhibition was rapidly reversed by readding calcium. Removing serosal calcium did not inhibit all active electrolyte transport processes, as the effect of a maximum concentration of theophylline (10 mM) was not altered. Similarly, d,l-verapamil, a calcium channel blocker, inhibited the serotonin-induced changes in short-circuit current and in net Na and net Cl fluxes, but did not alter the theophylline effects. In contrast, d-verapamil, a stereoisomer which does not block calcium channels, did not inhibit the serotonin-induced changes. The calcium dependence of these serotonin effects was associated with increased uptake of 45Ca into rabbit ileum, including increaed 45Ca uptake from the serosal surface. Serotonin also increased the rate of 45Ca efflux from rabbit ileum into a calcium-free solution, compatible with serotonin increasing the ileal plasma membrane permeability to calcium. It is postulated that serotonin affects active intestinal electrolyte transport by a mechanism dependent on serosal but not mucosal calcium that involves an increase in the intestinal plasma membrane permeability to calcium, and perhaps an increase in intracellular calcium.


Evaluation of the usability of various rapid antibody tests in the diagnostic application for COVID-19.
The usability of laboratory tests related to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critically important for the world undergoing the COVID-19 pandemic. The present study aimed to assess the diagnostic usability of rapid tests for the detection of antibody against SARS-CoV-2 through comparison of their results with the results of reverse transcription polymerase chain reaction (RT-PCR) test for the detection of SARS-CoV-2 genomic RNA and with the results of a quantitative test for antibody detection. Serum samples were collected from 18 patients undergoing RT-PCR testing for SARS-CoV-2. Twelve patients were RT-PCR positive while six were negative. A quantitative test based on chemiluminescent immunoassay and three rapid tests based on immunochromatography were performed to detect anti-SARS-CoV-2 IgG and IgM. All the antibody tests exhibited poor sensitivity at the timing of initial RT-PCR diagnosis. IgG responses occurring prior to or simultaneously with IgM responses were observed through not only the quantitative test but also the three rapid tests. Based on concordance with the quantitative test results, the large variance among the three rapid tests was revealed. All antibody tests were unsatisfactory to replace RT-PCR for the early diagnosis of COVID-19. Rapid antibody tests as well as a quantitative antibody test were useful in the assessment of immune responses in COVID-19. The obvious variance among the three rapid tests suggested limited accuracy and difficult standardization. Diagnostic usability of rapid antibody tests for COVID-19 should be investigated rigorously.


Immunological and virological benefits resulted from short-course treatment during primary HIV infection: a meta-analysis.
To assess the potential immunological and virological effects that result from short-course antiretroviral treatment during primary HIV infection (PHI). And to investigate whether treatment initiation time, treatment duration and follow-up time after treatment interruption would affect these post-treatment immunovirological outcomes. We systematically searched PubMed, Cochrane Library (to September 2013) and retrieved conference abstracts for studies regarding effects of early treatment during PHI on CD4 count and viral load (VL). Using the method of calculating weighted mean differences with Stata11.0, we conducted meta-analyses on the effect of early treatment on CD4 count and VL. Then we performed subgroup analyses by follow-up time after treatment interruption, treatment initiation time and treatment duration. Baseline immunovirological characteristics were also analyzed to account for potential bias. Compared to the untreated arm, treatment during PHI not only increased CD4 count by 85.92 cells/μl but also lowered viral load by 0.30 log copies/ml within one year after treatment interruption. However, the benefits declined gradually, reaching no significance 12-24 months after treatment interruption. Baseline immunovirological characteristics and sensitivity analyses of randomized controlled trials indicated that the benefits mentioned above were underestimated. Extending treatment duration beyond 12 months did not increase efficacy. Short-course treatment during PHI was associated with immunological and virological benefits which last for at least one year after treatment interruption. The conclusions from our study would help the decision-making in the clinical management of PHI.


The VMAT-2 inhibitor tetrabenazine affects effort-related decision making in a progressive ratio/chow feeding choice task: reversal with antidepressant drugs.
Behavioral activation is a fundamental feature of motivation, and organisms frequently make effort-related decisions based upon evaluations of reinforcement value and response costs. Furthermore, people with major depression and other disorders often show anergia, psychomotor retardation, fatigue, and alterations in effort-related decision making. Tasks measuring effort-based decision making can be used as animal models of the motivational symptoms of depression, and the present studies characterized the effort-related effects of the vesicular monoamine transport (VMAT-2) inhibitor tetrabenazine. Tetrabenazine induces depressive symptoms in humans, and also preferentially depletes dopamine (DA). Rats were assessed using a concurrent progressive ratio (PROG)/chow feeding task, in which they can either lever press on a PROG schedule for preferred high-carbohydrate food, or approach and consume a less-preferred lab chow that is freely available in the chamber. Previous work has shown that the DA antagonist haloperidol reduced PROG work output on this task, but did not reduce chow intake, effects that differed substantially from those of reinforcer devaluation or appetite suppressant drugs. The present work demonstrated that tetrabenazine produced an effort-related shift in responding on the PROG/chow procedure, reducing lever presses, highest ratio achieved and time spent responding, but not reducing chow intake. Similar effects were produced by administration of the subtype selective DA antagonists ecopipam (D1) and eticlopride (D2), but not by the cannabinoid CB1 receptor neutral antagonist and putative appetite suppressant AM 4413, which suppressed both lever pressing and chow intake. The adenosine A2A antagonist MSX-3, the antidepressant and catecholamine uptake inhibitor bupropion, and the MAO-B inhibitor deprenyl, all reversed the impairments induced by tetrabenazine. This work demonstrates the potential utility of the PROG/chow procedure as a rodent model of the effort-related deficits observed in depressed patients.


Pathogenicity of Vibrio penaeicida for white shrimp Litopenaeus vannamei: a cysteine protease-like exotoxin as a virulence factor.
The pathogenicity of Vibrio penaeicida Strains KH-1 and AM101, their culture-free supernatant (CFS), and their protein fraction obtained by 40% of ammonium sulfate precipitation (PFs40) were assessed in experimental challenges against juvenile Litopenaeus vannamei. Live Vibrio cells, CFS, and PFs40 from the AM101 strain produced a significantly higher mortality (p < 0.05) compared to the KH-1 strain. Toxicity and median lethal doses (LD50) of Fast Protein Liquid Chromatography (FPLC) products were evaluated on L. vannamei. The first FPLC fraction sample (A) from PFs40 of the AM101 strain displayed LD50 values of 1.68 and 5.61 microg protein ind.(-1), respectively. The second FPLC process from Fraction A showed a peak (A1) also with toxic effects to shrimp. PFs40, Fraction A, and Peak A1 showed a 38.5 kDa molecular band (SDS-PAGE), with activity on a gelatin protease zymogram. The lethal effect of PFs40 and Fraction A was inhibited by Proteinase K, CuCl2, E-64, and heat (60 and 100 degrees C) treatments, but was not inhibited by EDTA-Na2, aprotinin, and soy trypsin treatments. These results and the zymogram inhibition test suggest the presence of a cysteine protease-like proteinaceous exotoxin as a dominant protease, secreted by V. penaeicida Strain AM101.


POPULATION GENETICS OF NEUTRAL MUTATIONS IN EXPONENTIALLY GROWING CANCER CELL POPULATIONS.
In order to analyze data from cancer genome sequencing projects, we need to be able to distinguish causative, or "driver," mutations from "passenger" mutations that have no selective effect. Toward this end, we prove results concerning the frequency of neutural mutations in exponentially growing multitype branching processes that have been widely used in cancer modeling. Our results yield a simple new population genetics result for the site frequency spectrum of a sample from an exponentially growing population.


Does Stenting as a Bridge to Surgery in Left-Sided Colorectal Cancer Obstruction Really Worsen Oncological Outcomes?
Although self-expandable metal stents are used as a bridge to surgery in patients with colorectal cancer obstruction, their long-term oncological outcomes are unclear. The aim of this study was to investigate long-term oncological outcomes of self-expandable metal stents as a bridge to surgery (stent group) compared with direct surgery (direct operation group) in patients with left-sided colorectal cancer obstruction. This was a retrospective chart review. This study was conducted at a single tertiary academic center. Of 113 patients who underwent curative surgery for left-sided colorectal cancer obstruction at Asan Medical Center between 2005 and 2011, 42 underwent direct surgery and 71 underwent self-expandable metal stent insertion followed by elective surgery. After 1:1 propensity-score matching, 42 patients were enrolled in both groups, and their postsurgical outcomes were compared. The primary outcomes of this study were long-term oncological outcomes, including overall survival and recurrence-free survival of patients in both groups. Three- and 5-year overall survival rates were similar in the stent (87.0% and 71.0%) and direct operation (76.4% and 76.4%) groups (p = 0.931). Three- and 5-year recurrence-free survival rates were also similar in the stent (91.9% and 66.4%) and direct operation (81.2% and 71.2%) groups (p = 0.581), as were postsurgical complication rates (9.5% and 16.7%; p = 0.344). No patient in either group experienced a permanent stoma. This study was limited by its small patient numbers and retrospective nature. The long-term oncological outcomes of self-expandable metal stents as a bridge to surgery may not be inferior to those of direct surgery for left-sided colorectal cancer obstruction.


Isolation from a polyvalent antivenom of antibodies to a myotoxin in Bothrops asper snake venom.
Antibodies against Bothrops asper myotoxin were purified from a polyvalent antivenom by affinity chromatography. These antibodies neutralized most of the myotoxic activity of crude B. asper venom, as judged by histologic evaluation of skeletal muscle and determination of plasma creatine kinase levels in mice. When tested by immunoelectrophoresis, purified antibodies formed two superimposed bands of precipitation against an homogeneous (by SDS-PAGE analysis) preparation of B. asper myotoxin, as well as against crude B. asper venom. Ouchterlony immunodiffusion analysis of purified antibodies showed two precipitation bands with a pattern of complete immunologic identity between samples of crude B. asper venoms from specimens collected in the Atlantic and Pacific regions of Costa Rica. In addition, these antibodies precipitated when reacted against venom of newborn B. asper specimens from the Pacific region, but not against venom of newborn specimens from the Atlantic region. Purified antibodies were tested by immunodiffusion against eleven different snake venoms from Costa Rica. Only the venom of B. schlegelii cross-reacted, indicating the presence in this venom of components immunologically related to B. asper myotoxin. Analysis of purified antibodies to B. asper myotoxin by agarose electrophoresis and by SDS-PAGE suggests the presence of both IgG and IgM on the basis of electrophoretic position and molecular weight of the bands. Results obtained in neutralization experiments suggest that this myotoxin is a major factor in the development of local myonecrosis induced by crude B. asper venom.


How to Establish and Follow up a Large Prospective Cohort Study in the 21st Century--Lessons from UK COSMOS.
Large-scale prospective cohort studies are invaluable in epidemiology, but they are increasingly difficult and costly to establish and follow-up. More efficient methods for recruitment, data collection and follow-up are essential if such studies are to remain feasible with limited public and research funds. Here, we discuss how these challenges were addressed in the UK COSMOS cohort study where fixed budget and limited time frame necessitated new approaches to consent and recruitment between 2009-2012. Web-based e-consent and data collection should be considered in large scale observational studies, as they offer a streamlined experience which benefits both participants and researchers and save costs. Commercial providers of register and marketing data, smartphones, apps, email, social media, and the internet offer innovative possibilities for identifying, recruiting and following up cohorts. Using examples from UK COSMOS, this article sets out the dos and don'ts for today's cohort studies and provides a guide on how best to take advantage of new technologies and innovative methods to simplify logistics and minimise costs. Thus a more streamlined experience to the benefit of both research participants and researchers becomes achievable.


CD4 enumeration technologies: a systematic review of test performance for determining eligibility for antiretroviral therapy.
Measurement of CD4+ T-lymphocytes (CD4) is a crucial parameter in the management of HIV patients, particularly in determining eligibility to initiate antiretroviral treatment (ART). A number of technologies exist for CD4 enumeration, with considerable variation in cost, complexity, and operational requirements. We conducted a systematic review of the performance of technologies for CD4 enumeration. Studies were identified by searching electronic databases MEDLINE and EMBASE using a pre-defined search strategy. Data on test accuracy and precision included bias and limits of agreement with a reference standard, and misclassification probabilities around CD4 thresholds of 200 and 350 cells/μl over a clinically relevant range. The secondary outcome measure was test imprecision, expressed as % coefficient of variation. Thirty-two studies evaluating 15 CD4 technologies were included, of which less than half presented data on bias and misclassification compared to the same reference technology. At CD4 counts <350 cells/μl, bias ranged from -35.2 to +13.1 cells/μl while at counts >350 cells/μl, bias ranged from -70.7 to +47 cells/μl, compared to the BD FACSCount as a reference technology. Misclassification around the threshold of 350 cells/μl ranged from 1-29% for upward classification, resulting in under-treatment, and 7-68% for downward classification resulting in overtreatment. Less than half of these studies reported within laboratory precision or reproducibility of the CD4 values obtained. A wide range of bias and percent misclassification around treatment thresholds were reported on the CD4 enumeration technologies included in this review, with few studies reporting assay precision. The lack of standardised methodology on test evaluation, including the use of different reference standards, is a barrier to assessing relative assay performance and could hinder the introduction of new point-of-care assays in countries where they are most needed.


Bisphosphonates: safety and efficacy in the treatment and prevention of osteoporosis.
Osteoporosis affects more than 28 million Americans. With the advent of accessible and affordable diagnostic studies, awareness and recognition of this disease by patients and clinicians are growing. Osteoporotic fractures of the spine and hip are costly and associated with significant morbidity and mortality. Over the past decade, a surge of new antiosteoporotic drugs have been labeled or are awaiting labeling by the U.S. Food and Drug Administration. One class of agents used to treat osteoporosis is the bisphosphonates, which inhibit bone resorption, cause an increase in bone mineral density and reduce the risk of future fractures caused by aging, estrogen deficiency and corticosteroid use. Overall, bisphosphonates have been shown to have a strong safety and tolerability profile.


Genome screen to identify susceptibility genes for Parkinson disease in a sample without parkin mutations.
Parkinson disease (PD) is a common neurodegenerative disorder characterized by bradykinesia, resting tremor, muscular rigidity, and postural instability, as well as by a clinically significant response to treatment with levodopa. Mutations in the alpha-synuclein gene have been found to result in autosomal dominant PD, and mutations in the parkin gene produce autosomal recessive juvenile-onset PD. We have studied 203 sibling pairs with PD who were evaluated by a rigorous neurological assessment based on (a) inclusion criteria consisting of clinical features highly associated with autopsy-confirmed PD and (b) exclusion criteria highly associated with other, non-PD pathological diagnoses. Families with positive LOD scores for a marker in an intron of the parkin gene were prioritized for parkin-gene testing, and mutations in the parkin gene were identified in 22 families. To reduce genetic heterogeneity, these families were not included in subsequent genome-screen analysis. Thus, a total of 160 multiplex families without evidence of a parkin mutation were used in multipoint nonparametric linkage analysis to identify PD-susceptibility genes. Two models of PD affection status were considered: model I included only those individuals with a more stringent diagnosis of verified PD (96 sibling pairs from 90 families), whereas model II included all examined individuals as affected, regardless of their final diagnostic classification (170 sibling pairs from 160 families). Under model I, the highest LOD scores were observed on chromosome X (LOD score 2.1) and on chromosome 2 (LOD score 1.9). Analyses performed with all available sibling pairs (model II) found even greater evidence of linkage to chromosome X (LOD score 2.7) and to chromosome 2 (LOD score 2.5). Evidence of linkage was also found to chromosomes 4, 5, and 13 (LOD scores >1.5). Our findings are consistent with those of other linkage studies that have reported linkage to chromosomes 5 and X.


The effects of temporally variable dispersal and landscape structure on invasive species spread.
Many invasive species are too widespread to realistically eradicate. For such species, a viable management strategy is to slow the rate of spread. However, to be effective, this will require detailed spread data and an understanding of the influence of environmental conditions and landscape structure on invasion rates. We used a time series of remotely sensed distribution maps and a spatial simulation model to study spread of the invasive Lepidium latifolium (perennial pepperweed) in California's Sacramento-San Joaquin River Delta. L. latifolium is a noxious weed and exhibited rapid, explosive spread. Annual infested area and empirical dispersal kernels were derived from the remotely sensed distributions in order to assess the influence of weather conditions on spread and to parameterize the simulation model. Spread rates and dispersal distances were highest for nascent infestations and in years with wet springs. Simulations revealed that spread rates were more strongly influenced by the length of long-distance dispersal than by temporal variation in its likelihood. It is thus important to capture long-distance dispersal and the conditions that facilitate spread when collecting data to parameterize spread models. Additionally, management actions performed in high-spread years, targeting long-distance recruits, can effectively contain infestations. Corridors were relatively unimportant to spread rates; their effectiveness at enhancing rate of spread was limited by the species' dispersal ability and the time needed to travel through the corridor. In contrast, habitat abundance and shape surrounding the introduction site strongly influenced invasion dynamics. Satellite patches invading large areas of invasible habitat present especially high risk.