Combined use of Ovsynch and progesterone supplementation after artificial insemination in dairy cattle.
The objective of this study was to evaluate the effects of different Ovsynch protocols combined with progesterone (P4) supplementation after artificial insemination (AI) of Holstein-Friesian cows. Cows were randomly synchronized at 52 to 63 d after parturition with either the classical Ovsynch protocol (GnRH on d 0, PGF(2α) on d 7, GnRH 48 h after PGF(2α)) or with a modified Ovsynch protocol (second GnRH 60 h after PGF(2α)). On d 4 after timed AI (TAI), the cows were blocked by parity and randomly divided into 2 groups. Half of the cows were supplemented with P4 (P4+) by applying a P4-releasing intravaginal device intravaginally for 14 d, whereas the other half remained untreated (P4-). In 50% of randomly chosen cows, plasma P4 was measured on d 4, 5, and 18 after TAI. Sonographic pregnancy diagnosis was performed on d 33 after TAI in a total of 398 cows. Health status and body condition score (BCS) of all cows were examined at several stages of the study. Cows in the modified Ovsynch protocol tended to have higher P4 values on d 4 after TAI than cows in the classical Ovsynch protocol (2.1 ± 0.2 vs. 1.6 ± 0.2 ng/mL), but no difference in pregnancy per AI (P/AI) was observed between the 2 Ovsynch protocols (38.4% vs. 44.1%). Independent of the Ovsynch protocols, P4+ cows tended to have higher P/AI compared with P4- cows (44.4% vs. 38.1%). The retention of fetal membranes and BCS at the time of insemination affected P/AI. Moreover, an interaction between BCS at the time of insemination and P4 supplementation was apparent; that is, the difference in P/AI between P4+ and P4- cows was significant in cows with BCS ≥3.25. Progesterone-supplemented cows showed higher P4 values on d 5 (4.9 ± 0.2 vs. 2.6 ± 0.2) and d 18 (7.8 ± 0.2 vs. 6.3 ± 0.2) after TAI, respectively. In conclusion, the elongation of the time interval between the injections of PGF(2α) and the second GnRH from 48 to 60 h had no effect on P/AI. Progesterone supplementation after insemination improved the P/AI of the Ovsynch protocols, but this effect was more apparent in cows with BCS ≥3.25.


Efficacy and safety of adjunctive armodafinil in adults with major depressive episodes associated with bipolar I disorder: a randomized, double-blind, placebo-controlled, multicenter trial.
To examine the efficacy and safety of adjunctive armodafinil for major depressive episodes associated with bipolar I disorder. Adults meeting DSM-IV-TR criteria for bipolar I disorder and currently experiencing a major depressive episode while taking at least 4 weeks of conventional maintenance medication were enrolled in a placebo-controlled evaluation of adjunctive armodafinil 150 or 200 mg (conducted January 2010-March 2012). The primary efficacy measure was change from baseline to week 8 on the 30-item Inventory of Depressive Symptomatology-Clinician-Rated (IDS-C30) total score in the 150-mg armodafinil group versus placebo. Of 786 patients screened, 433 were randomized (placebo, n = 199; armodafinil 150 mg, n = 201; armodafinil 200 mg, n = 33). The 200-mg armodafinil group was discontinued by protocol amendment due to lower than expected patient enrollment. For the 150-mg armodafinil group versus placebo, there was a significantly greater decrease in least squares mean (standard error of mean [SEM]) IDS-C30 total score at week 8 (-21.7 [1.1] vs -17.9 [1.1]; P = .0097; Cohen d therapeutic effect size = 0.28). The proportion of IDS-C30 responders (≥ 50% decrease from baseline) was significantly higher for the 150-mg armodafinil group versus placebo at final visit (46% [91/197] vs 34% [67/196]; P = .0147). The proportion of IDS-C30 remitters (total score ≤ 11) was 21% (42/197) for armodafinil 150 mg versus 17% (34/196) for placebo (P = .3343) at final visit. Adverse events (AEs) observed in > 5% of either the armodafinil 150 mg or placebo groups and more frequently with 150 mg armodafinil were diarrhea (9% [17/198] vs 7% [13/199]), and nausea (6% [11/198] vs 5% [9/199]), respectively. In the 200-mg armodafinil group, there were 2 serious AEs (n = 1, hepatic failure leading to death; n = 1, acute hepatitis). The death was not considered related to study treatment. Adjunctive armodafinil 150 mg significantly improved symptoms of major depressive episodes associated with bipolar I disorder versus placebo and was generally well tolerated. ClinicalTrials.gov identifier: NCT01072929.


Protected areas and regional avian species richness in South Africa.
Protected areas are generally regarded as essential for the long-term maintenance of biodiversity. Evidence for their effectiveness in this regard is, however, somewhat equivocal. Here, we document the relationship between the proportion of protected land and species richness in a region, both with and without taking spatial variation in environmental energy availability into account. Using the South African avifauna as a case study, we find that total and threatened species richness exhibit modest increases with the proportion of protected land. While the protected area network should be expanded, it is essential that conservation efforts also focus on maintaining biodiversity in the wider unprotected landscape that supports high species richness.


The Effects of Previous Error and Success in Alzheimer's Disease and Mild Cognitive Impairment.
This work investigated in Alzheimer's disease dementia (AD), whether the probability of making an error on a task (or a correct response) was influenced by the outcome of the previous trials. We used the antisaccade task (AST) as a model task given the emerging consensus that it provides a promising sensitive and early biological test of cognitive impairment in AD. It can be employed equally well in healthy young and old adults, and in clinical populations. This study examined eye-movements in a sample of 202 participants (42 with dementia due to AD; 65 with mild cognitive impairment (MCI); 95 control participants). The findings revealed an overall increase in the frequency of AST errors in AD and MCI compared to the control group, as predicted. The errors on the current trial increased in proportion to the number of consecutive errors on the previous trials. Interestingly, the probability of errors was reduced on the trials that followed a previously corrected error, compared to the trials where the error remained uncorrected, revealing a level of adaptive control in participants with MCI or AD dementia. There was an earlier peak in the AST distribution of the saccadic reaction times for the inhibitory errors in comparison to the correct saccades. These findings revealed that the inhibitory errors of the past have a negative effect on the future performance of healthy adults as well as people with a neurodegenerative cognitive impairment.


Nitric oxide mediates angiogenesis in vivo and endothelial cell growth and migration in vitro promoted by substance P.
We evaluated the effects of nitric oxide (NO) generators and endogenous production of NO elicited by substance P (SP) in the angiogenesis process. Angiogenesis was monitored in the rabbit cornea in vivo and in vitro by measuring the growth and migration of endothelial cells isolated from coronary postcapillary venules. The angiogenesis promoted in the rabbit cornea by [Sar9]-SP-sulfone, a stable and selective agonist for the tachykinin NK1 receptor, and by prostaglandin E1 (PGE1), was potentiated by sodium nitroprusside (SNP). Conversely, the NO synthase inhibitor N omega-nitro-L-arginine methyl ester (L-NAME), given systemically, inhibited angiogenesis elicited by [Sar9]-SP-sulfone and by PGE1. Endothelial cells exposed to SNP exhibited an increase in thymidine incorporation and in total cell number. Exposure of the cells to NO generating drugs, such as SNP, isosorbide dinitrate, and glyceryl trinitrate, produced a dose-dependent increase in endothelial cell migration. Capillary endothelial cell proliferation and migration produced by SP were abolished by pretreatment with the NO synthase inhibitors N omega-mono-methyl-L-arginine (L-NMMA), N omega-nitro-L-arginine (L-NNA), and L-NAME. Exposure of the cells to SP activated the calcium-dependent NO synthase. Angiogenesis and endothelial cell growth and migration induced by basic fibroblast growth factor were not affected by NO synthase inhibitors. These data indicate that NO production induced by vasoactive agents, such as SP, functions as an autocrine regulator of the microvascular events necessary for neovascularization and mediates angiogenesis.


Factors associated with seven-year incidence of diabetes complications among older Mexican Americans.
Diabetes is common in elderly persons and is highly prevalent in Mexican Americans. Little is known about factors associated with the incidence of diabetes complications. To examine the social, demographic and health factors associated with a 7-year incidence of diabetes complications among older Mexican Americans with diabetes. A 7-year prospective cohort of 536 non-institutionalized Mexican Americans aged > or =65 years with diabetes residing in the Southwest of USA. Measures included socio-demographic factors, duration of diabetes, diabetes treatment, medical conditions and body mass index. Diabetes complications were assessed by self-reports of any kidney, eye and circulation problems or amputation due to diabetes over a 7-year period. Subjects with disease duration of > or =10 years were found to have an increased incidence of kidney, eye and circulation problems or amputation over a 7-year period. Hazard ratio (HR) for circulation problems was significantly higher in foreign-born subjects with >15 years in the USA. Foreign-born subjects with <15 years in the USA and those with a frequent number of physician visits were at increased risk of kidney complications. Subjects with baseline eye complications had a higher HR for incidence of amputations in the next 7 years. Subjects with baseline circulation complications had a higher incidence of amputations, eye and kidney complications in the next 7 years. Our data suggest that risks of diabetes complications are influenced by different factors. Prolonged diabetes duration, being foreign-born, living in the USA for a longer period, and frequent physician visits were factors associated with diabetes complications. Designing intervention strategies to reduce diabetes complication risks requires an understanding of the differences in demographic and health predictors of such risks.


Iron phthalocyanine in non-aqueous medium forming layer-by-layer films: growth mechanism, molecular architecture and applications.
The application of organic thin films as transducer elements in electronic devices has been widely exploited, with the electrostatic layer-by-layer (LbL) technique being one of the most powerful tools to produce such films. The conventional LbL method, however, is restricted in many cases to water soluble compounds. Here, an alternative way to produce LbL films containing iron phthalocyanine (FePc) in non-aqueous media (chloroform) is presented. This film fabrication was made possible by exploiting the specific interactions between Fe and NH(2) groups from PAH, poly(allylamine hydrochloride) used as the supporting layer, leading to the formation of bilayers structured as (PAH/FePc)(n). We have also incorporated silver nanoparticles (AgNPs) in LbL films with (PAH/FePc/AgNP)(n) trilayers, making it possible to achieve the surface-enhanced Raman scattering (SERS) phenomenon. The molecular architecture of the LbL films was determined through different techniques. The growth was monitored with UV-Vis absorption spectroscopy, their morphology characterized by optical and scanning electron (SEM) microscopes, and their molecular organization determined using FTIR. The electrochemical properties of the LbL films were successfully applied in detecting dopamine in KCl aqueous solutions at different concentrations using cyclic voltammetry. The results confirmed that the LbL films from FePc in non-aqueous media keep their electroactivity, while showing an interesting electrocatalytic effect. The SERS phenomenon suggested that FePc aggregates might be directly involved in the maintenance of the electroactivity of the LbL films.


An antifungal polyketide associated with horizontally acquired genes supports symbiont-mediated defense in Lagria villosa beetles.
Microbial symbionts are often a source of chemical novelty and can contribute to host defense against antagonists. However, the ecological relevance of chemical mediators remains unclear for most systems. Lagria beetles live in symbiosis with multiple strains of Burkholderia bacteria that protect their offspring against pathogens. Here, we describe the antifungal polyketide lagriamide, and provide evidence supporting that it is produced by an uncultured symbiont, Burkholderia gladioli Lv-StB, which is dominant in field-collected Lagria villosa. Interestingly, lagriamide is structurally similar to bistramides, defensive compounds found in marine tunicates. We identify a gene cluster that is probably involved in lagriamide biosynthesis, provide evidence for horizontal acquisition of these genes, and show that the naturally occurring symbiont strains on the egg are protective in the soil environment. Our findings highlight the potential of microbial symbionts and horizontal gene transfer as influential sources of ecological innovation.


The Society of Thoracic Surgeons Congenital Heart Surgery Database: 2018 Update on Outcomes and Quality.
The Society of Thoracic Surgeons Congenital Heart Surgery Database is a comprehensive registry of clinical outcomes that captures almost all pediatric cardiac surgical operations in the United States. It is the platform for all activities of The Society of Thoracic Surgeons related to the analysis of outcomes and the improvement of quality in this subspecialty. This article summarizes current aggregate national outcomes in congenital and pediatric cardiac surgery and reviews related activities in the areas of quality measurement, performance improvement, and transparency. The reported data about aggregate national outcomes are exemplified by an analysis of 10 benchmark operation groups performed from January 2013 through December 2016. This analysis documents the overall aggregate Operative Mortality (interquartile range among all participating programs) for the following procedural groups: off-bypass coarctation repair, 1.3% (0.0% to 1.4%%); ventricular septal defect repair, 0.6% (0.0% to 0.9%); tetralogy of Fallot repair, 1.1% (0.0% to 2.0%); complete atrioventricular canal repair, 2.7% (0.0% to 4.4%); arterial switch operation, 2.2% (0.0% to 2.9%); arterial switch operation and ventricular septal defect repair, 5.1% (0.0% to 8.3%); Glenn/HemiFontan, 2.1% (0.0% to 3.1%); Fontan operation, 1.1% (0.0% to 0.0%); truncus arteriosus repair, 10.1% (0.0% to 15.4%); and Norwood procedure, 15.8% (9.0% to 25.0%).


Flavonoid-rich cocoa consumption affects multiple cardiovascular risk factors in a meta-analysis of short-term studies.
A growing body of evidence suggests that the consumption of foods rich in polyphenolic compounds, particularly cocoa, may have cardioprotective effects. No review, however, has yet examined the effect of flavonoid-rich cocoa (FRC) on all major cardiovascular risk factors or has examined potential dose-response relationships for these effects. A systematic review and meta-analysis of randomized, controlled trials was performed to evaluate the effect of FRC on cardiovascular risk factors and to assess a dose-response relationship. Inclusion and exclusion criteria as well as dependent and independent variables were determined a priori. Data were collected for: blood pressure, pulse, total cholesterol, HDL cholesterol, LDL cholesterol, TG, BMI, C-reactive protein, flow-mediated vascular dilation (FMD), fasting glucose, fasting insulin, serum isoprostane, and insulin sensitivity/resistance indices. Twenty-four papers, with 1106 participants, met the criteria for final analysis. In response to FRC consumption, systolic blood pressure decreased by 1.63 mm Hg (P = 0.033), LDL cholesterol decreased by 0.077 mmol/L (P = 0.038), and HDL cholesterol increased by 0.046 mmol/L (P = 0.037), whereas total cholesterol, TG, and C-reactive protein remained the same. Moreover, insulin resistance decreased (HOMA-IR: -0.94 points; P < 0.001), whereas FMD increased (1.53%; P < 0.001). A nonlinear dose-response relationship was found between FRC and FMD (P = 0.004), with maximum effect observed at a flavonoid dose of 500 mg/d; a similar relationship may exist with HDL cholesterol levels (P = 0.06). FRC consumption significantly improves blood pressure, insulin resistance, lipid profiles, and FMD. These short-term benefits warrant larger long-term investigations into the cardioprotective role of FRC.


Impacts of caring for a child with the CDKL5 disorder on parental wellbeing and family quality of life.
Although research in this area remains sparse, raising a child with some genetic disorders has been shown to adversely impact maternal health and family quality of life. The aim of this study was to investigate such impacts in families with a child with the CDKL5 disorder, a newly recognised genetic disorder causing severe neurodevelopmental impairments and refractory epilepsy. Data were sourced from the International CDKL5 Disorder Database to which 192 families with a child with a pathogenic CDKL5 mutation had provided data by January 2016. The Short Form 12 Health Survey Version 2, yielding a Physical Component Summary and a Mental Component Summary score, was used to measure primary caregiver's wellbeing. The Beach Center Family Quality of Life Scale was used to measure family quality of life. Linear regression analyses were used to investigate relationships between child and family factors and the various subscale scores. The median (range) age of the primary caregivers was 37.0 (24.6-63.7) years and of the children was 5.2 (0.2-34.1) years. The mean (SD) physical and mental component scores were 53.7 (8.6) and 41.9 (11.6), respectively. In mothers aged 25-54 years the mean mental but not the physical component score was lower than population norms. After covariate adjustment, caregivers with a tube-fed child had lower mean physical but higher mean mental component scores than those whose child fed orally (coefficient = -4.80 and 6.79; p = 0.009 and 0.012, respectively). Child sleep disturbances and financial hardship were negatively associated with the mental component score. The mean (SD) Beach Center Family Quality of Life score was 4.06 (0.66) and those who had used respite services had lower scores than those who had not across the subscales. Emotional wellbeing was considerably impaired in this caregiver population, and was particularly associated with increased severity of child sleep problems and family financial difficulties. Family quality of life was generally rated lowest in those using respite care extensively, suggesting that these families may be more burdened by daily caregiving.


Sample size considerations for stratified cluster randomization design with binary outcomes and varying cluster size.
Stratified cluster randomization trials (CRTs) have been frequently employed in clinical and healthcare research. Comparing with simple randomized CRTs, stratified CRTs reduce the imbalance of baseline prognostic factors among different intervention groups. Due to the popularity, there has been a growing interest in methodological development on sample size estimation and power analysis for stratified CRTs; however, existing work mostly assumes equal cluster size within each stratum and uses multilevel models. Clusters are often naturally formed with random sizes in CRTs. With varying cluster size, commonly used ad hoc approaches ignore the variability in cluster size, which may underestimate (overestimate) the required number of clusters for each group per stratum and lead to underpowered (overpowered) clinical trials. We propose closed-form sample size formulas for estimating the required total number of subjects and for estimating the number of clusters for each group per stratum, based on Cochran-Mantel-Haenszel statistic for stratified cluster randomization design with binary outcomes, accounting for both clustering and varying cluster size. We investigate the impact of various design parameters on the relative change in the required number of clusters for each group per stratum due to varying cluster size. Simulation studies are conducted to evaluate the finite-sample performance of the proposed sample size method. A real application example of a pragmatic stratified CRT of a triad of chronic kidney disease, diabetes, and hypertension is presented for illustration.


Unusual binding interactions in PDZ domain crystal structures help explain binding mechanisms.
PDZ domains most commonly bind the C-terminus of their protein targets. Typically the C-terminal four residues of the protein target are considered as the binding motif, particularly the C-terminal residue (P0) and third-last residue (P-2) that form the major contacts with the PDZ domain's "binding groove". We solved crystal structures of seven human PDZ domains, including five of the seven PDLIM family members. The structures of GRASP, PDLIM2, PDLIM5, and PDLIM7 show a binding mode with only the C-terminal P0 residue bound in the binding groove. Importantly, in some cases, the P-2 residue formed interactions outside of the binding groove, providing insight into the influence of residues remote from the binding groove on selectivity. In the GRASP structure, we observed both canonical and noncanonical binding in the two molecules present in the asymmetric unit making a direct comparison of these binding modes possible. In addition, structures of the PDZ domains from PDLIM1 and PDLIM4 also presented here allow comparison with canonical binding for the PDLIM PDZ domain family. Although influenced by crystal packing arrangements, the structures nevertheless show that changes in the positions of PDZ domain side-chains and the alpha B helix allow noncanonical binding interactions. These interactions may be indicative of intermediate states between unbound and fully bound PDZ domain and target protein. The noncanonical "perpendicular" binding observed potentially represents the general form of a kinetic intermediate. Comparison with canonical binding suggests that the rearrangement during binding involves both the PDZ domain and its ligand.


The effect of dietary intake of coenzyme Q10 on skin parameters and condition: Results of a randomised, placebo-controlled, double-blind study.
Coenzyme Q10 (CoQ10) is a natural constituent of foods and is also often used in both functional foods and supplements. In addition, it is a common ingredient of cosmetics where it is believed to reduce the signs of skin ageing. However, the existing data about the effect of dietary intake of CoQ10 on skin parameters and condition are scarce. To gain an insight into this issue, we conducted a double-blind, placebo-controlled experiment with 33 healthy subjects. Our objective was to investigate the effects of 12 weeks of daily supplementation with 50 and 150 mg of CoQ10 on skin parameters and condition. Study was conducted with a water-soluble form of CoQ10 with superior bioavailability (Q10Vital® ). While the results of some previous in vitro studies showed possible protection in UVB response, we did not observe significant changes in the minimal erythema dose (MED). On the other hand, the intake of CoQ10 limited seasonal deterioration of viscoelasticity and reduced some visible signs of ageing. We determined significantly reduced wrinkles and microrelief lines, and improved skin smoothness. Supplementation with CoQ10 did not significantly affect skin hydration and dermis thickness. © 2016 BioFactors, 43(1):132-140, 2017.


Bolus tracking for improved metabolic imaging of hyperpolarised compounds.
Dynamic nuclear polarisation has enabled real-time metabolic imaging of pyruvate and its metabolites. Conventional imaging sequences rely on predefined settings and do not account for intersubject variations in biological parameters such as perfusion. We present a fully automatic real-time bolus tracking sequence for hyperpolarised substrates which starts the imaging acquisition at a defined point on the bolus curve. This reduces artefacts due to signal change and allows for a more efficient use of hyperpolarised magnetisation. For single time point imaging methods, bolus tracking enables a more reliable and consistent quantification of metabolic activity. An RF excitation with a small flip angle is used to obtain slice-selective pyruvate tracking information in rats. Moreover, in combination with a copolarised urea and pyruvate injection, spectrally selective tracking on urea allows obtaining localised bolus tracking information without depleting the pyruvate signal. Particularly with regard to clinical application, the bolus tracking technique could provide an important step towards a routine assessment protocol which removes operator dependencies and ensures comparable results.


Environmentally Mediated Social Dilemmas.
By consuming and producing environmental resources, organisms inevitably change their habitats. The consequences of such environmental modifications can be detrimental or beneficial not only to the focal organism but also to other organisms sharing the same environment. Social evolution theory has been very influential in studying how social interactions mediated by public 'goods' or 'bads' evolve by emphasizing the role of spatial structure. The environmental dimensions driving these interactions, however, are typically abstracted away. We propose here a new, environment-mediated taxonomy of social behaviors where organisms are categorized by their production or consumption of environmental factors that can help or harm others in the environment. We discuss microbial examples of our classification and highlight the importance of environmental intermediates more generally.


Deformation responses of a physically cross-linked high molecular weight elastin-like protein polymer.
Recombinant protein polymers were synthesized and examined under various loading conditions to assess the mechanical stability and deformation responses of physically cross-linked, hydrated, protein polymer networks designed as triblock copolymers with central elastomeric and flanking plastic-like blocks. Uniaxial stress-strain properties, creep and stress relaxation behavior, as well as the effect of various mechanical preconditioning protocols on these responses were characterized. Significantly, we demonstrate for the first time that ABA triblock protein copolymers when redesigned with substantially larger endblock segments can withstand significantly greater loads. Furthermore, the presence of three distinct phases of deformation behavior was revealed upon subjecting physically cross-linked protein networks to step and cyclic loading protocols in which the magnitude of the imposed stress was incrementally increased over time. We speculate that these phases correspond to the stretch of polypeptide bonds, the conformational changes of polypeptide chains, and the disruption of physical cross-links. The capacity to select a genetically engineered protein polymer that is suitable for its intended application requires an appreciation of its viscoelastic characteristics and the capacity of both molecular structure and conditioning protocols to influence these properties.


Differential methylation of a short CpG-rich sequence within exon 1 of TCF21 gene: a promising cancer biomarker assay.
Detection of cancer cells at early stages could potentially increase survival rates in cancer patients. Aberrant promoter hypermethylation is a major mechanism for silencing tumor suppressor genes in many kinds of human cancers. A recent report from our laboratory described the use of quantitative methylation-specific PCR assays for discriminating patients with lung cancer from those without lung cancer using lung biopsies as well as sputum samples. TCF21 is known to be essential for differentiation of epithelial cells adjacent to mesenchyme. Using restriction landmark genomic scanning, a recent study identified TCF21 as candidate tumor suppressor at 6q23-q24 that is epigenetically inactivated in lung and head and neck cancers. Using DNA sequencing technique, we narrowed down a short CpG-rich segment (eight specific CpG sites in the CpG island within exon 1) of the TCF21 gene, which was unmethylated in normal lung epithelial cells but predominantly methylated in lung cancer cell lines. We specifically targeted this short CpG-rich sequence and developed a quantitative methylation-specific PCR assay suitable for high-throughput analysis. We showed the usefulness of this assay in discriminating patients with lung cancer from those without lung cancer using biopsies and sputum samples. We further showed similar applications with multiple other malignancies. Our assay might have important implications in early detection and surveillance of multiple malignancies.


Effects of high dose inhaled beclomethasone dipropionate, 750 micrograms and 1500 micrograms twice daily, and 40 mg per day oral prednisolone on lung function, symptoms, and bronchial hyperresponsiveness in patients with non-asthmatic chronic airflow obstruction.
The effect of treatment with inhaled corticosteroids in patients with non-asthmatic chronic airflow obstruction is still disputed. Whether any physiological improvements seen are accompanied by changes in bronchial responsiveness and symptoms and quality of life is also still unclear. A sequential placebo controlled, blinded parallel group study investigating the effect of three weeks of treatment with inhaled beclomethasone dipropionate (BDP), 750 micrograms or 1500 micrograms twice daily, and oral prednisolone, 40 mg per day, was carried out in 105 patients with severe non-asthmatic chronic airflow obstruction (mean age 66 years, mean forced expiratory volume in one second (FEV1) 1.05 litres [40% predicted], geometric mean PD20 0.52 mumol). End points assessed were FEV1, forced vital capacity (FVC), and peak expiratory flow (PEF), bronchial responsiveness to inhaled histamine, and quality of life as measured by a formal quality of life questionnaire. Both doses of BDP produced equivalent, small, but significant improvements in FEV1 (mean 48 ml), FVC (mean 120 ml), and PEF (mean 12.4 l/min). The addition of oral prednisolone to the treatment regime in two thirds of the patients did not produce any further improvement in these parameters. Inhaled BDP produced a treatment response in individual patients (defined as an improvement in FEV1, FVC, or mean PEF of at least 20% compared with baseline values) more commonly than placebo (34% v 15%). The two doses of BDP were equally effective in this respect and again no further benefit of treatment with oral prednisolone was noted. Treatment with BDP for up to six weeks did not affect bronchial responsiveness to histamine. Small but significant improvements were seen in dyspnoea during daily activities, and the feeling of mastery over the disease. High dose inhaled BDP is an effective treatment for patients with chronic airflow obstruction not caused by asthma. Both objective and subjective measures show improvement. Unlike asthma, no improvement in bronchial responsiveness was detected after six weeks of treatment.


Loss of cargo binding in the human myosin VI deafness mutant (R1166X) leads to increased actin filament binding.
Mutations in myosin VI have been associated with autosomal-recessive (DFNB37) and autosomal-dominant (DFNA22) deafness in humans. Here, we characterise an myosin VI nonsense mutation (R1166X) that was identified in a family with hereditary hearing loss in Pakistan. This mutation leads to the deletion of the C-terminal 120 amino acids of the myosin VI cargo-binding domain, which includes the WWY-binding motif for the adaptor proteins LMTK2, Tom1 as well as Dab2. Interestingly, compromising myosin VI vesicle-binding ability by expressing myosin VI with the R1166X mutation or with single point mutations in the adaptor-binding sites leads to increased F-actin binding of this myosin in vitro and in vivo As our results highlight the importance of cargo attachment for regulating actin binding to the motor domain, we perform a detailed characterisation of adaptor protein binding and identify single amino acids within myosin VI required for binding to cargo adaptors. We not only show that the adaptor proteins can directly interact with the cargo-binding tail of myosin VI, but our in vitro studies also suggest that multiple adaptor proteins can bind simultaneously to non-overlapping sites in the myosin VI tail. In conclusion, our characterisation of the human myosin VI deafness mutant (R1166X) suggests that defects in cargo binding may leave myosin VI in a primed/activated state with an increased actin-binding ability.


Cruising for truckers on highways and the internet: sexual networks and infection risk.
Empirical evidence on the heterosexual partnerships of long-haul truckers suggests connections among occupational stressors, substance misuse, structural factors, and risk for sexually transmitted infections and HIV. Yet the potential risks associated with same-sex partnerships of truckers and truckchasers (men who specifically cruise for truckers) remain largely unknown. Drawing from diverse sources as well as primary and secondary data from 173 truckers and "truckchasers," we discuss how trucking and cruising contexts, in conjunction with Internet fora, jointly create a risk-enabling environment for truckers and their sex contacts. Findings point toward an elusive but extensive sexual network that spans across the Internet and highways and takes advantage of truckers' mobility as it bridges disparate epidemiological spaces and populations. The delineation of cruising within the hypermasculine occupational sector of trucking adds new insights to the study of sexual health, which is particularly important considering the riskladen sex contacts of truckers and truckchasers and potential for infection spread.


Salt and Health: Survey on Knowledge and Salt Intake Related Behaviour in Italy.
Excess sodium intake is a recognised causal factor of hypertension and its cardiovascular complications; there is however a lack of practical instruments to assess and monitor the level of knowledge and behaviour about dietary salt intake and to relate these factors to the population general dietary habits. A self-administered questionnaire was developed to assess the salt and health related knowledge and behaviour of the Italian population through an online survey. A sample of 11,618 Italian participants completed the questionnaire. The degree of knowledge and the reported behaviour about salt intake were both found to be related to age, gender, home region, level of education and occupation. There was a significant interrelation between salt knowledge and behaviour and both were significantly and directly related to the degree of adherence to a Mediterranean-like dietary pattern. A hierarchical evaluation was also made of the relevance of any single question to the overall assessment of knowledge and behaviour about salt intake. The study population overall appeared to have a decent level of knowledge about salt, but a less satisfactory behaviour. Our findings point to social inequalities and young age as the main factors having a negative impact on knowledge and behaviour about salt intake as part of generally inadequate dietary habits. The degrees of knowledge and behaviour were significantly and directly interrelated, confirming that improving knowledge is a key step for behavioural changes, and suggesting that educational campaigns are crucial for the implementation of good practices in nutrition.


Using the Morbidity and Mortality conference to teach and assess the ACGME General Competencies.
The weekly Morbidity and Mortality (M&M) conference, a Residency Review Committee on Surgery required conference, is a hallmark of general surgery residency training. This conference has been used traditionally to teach and assess the ACGME General Competencies of patient care and medical knowledge. The author's department has changed the format of their weekly M&M conference so that it enables them to teach and assess residents also in terms of the ACGME General Competencies of practice-based learning and improvement, professionalism, interpersonal and communication skills, and systems-based practice. Each Monday the chief resident on each teaching service compiles a list of patient discharges and deaths for the previous week. Although all deaths are presented, only significant patient complications are selected for the following week's M&M conference. This 2-week preparation period enables the resident, who was primarily involved in the care of the patient, to thoroughly review the case and prepare his/her presentation. At the conference, the resident presents the patient's history and discusses the complication or death, not only in terms of the patient care provided (traditional M&M model), but also it analyzes the case in terms of health-care systems problems that may have contributed to the patient's morbidity and/or mortality; patient safety issues; communication problems with the patient, family, or other health-care workers; and ethnic or ethical issues related to the care provided. The case is then reviewed by faculty surgeons. Again, not only is the patient care critiqued, but also systems problems, communication problems, and ethical dilemmas. Each resident who presents a case at the M&M conference also completes a practice-based improvement log. This form analyzes the patient's outcome including factors leading to the complication and/or mortality, opportunities for systems improvement, patient safety or communication problems, ethnic or ethical issues, what the resident would do different in his/her practice, and references consulted for this case. These forms are reviewed with the resident by the Residency Director and become part of the resident's portfolio. The restructuring of the M&M conference so that a case is analyzed with all ACGME General Competencies has made the M&M conference more interesting and has improved the educational aspects of the conference. Analyzing a case according to the various ACGME General Competencies has provided another method to teach these competencies to their residents and a tool to determine whether the residents are meeting the competencies.


Kitona 2013: A Medical Readiness Training Exercise in the Democratic Republic of the Congo Leading to Lion Rouge.
Health initiatives support regional stability and are a priority for US and African partners. We present data and experience from the Democratic Republic of Congo (DRC), a strategically and epidemiologically ideal location for collaborative medical engagement (CME). Our objectives included relationship building, exposure of US military medical personnel to uncommon tropical diseases, bolstering a referral hospital, and updating Congolese physicians on new treatment or preventive standards of care. We conducted a CME-styled medical readiness training exercise (MEDRETE) at the Military Referral Hospital of Kitona in June 2013. US and Congolese healthcare providers presented 20 lectures and evaluated 158 patients collaboratively; 132 for infections. The CME led to Lion Rouge, the first joint military, multidisciplinary engagement between the respective militaries. Equally noteworthy is that some of the same participants returned to the same location for the follow-on exercise, providing continuity. These outcomes suggest the MEDRETE and CME approaches were successful.


Patient-reported outcome measures for adult chronic rhinosinusitis: A systematic review and quality assessment.
With a focus on patient-centered care, there is increasing policy interest in patient-reported outcome measures (PROMs) to inform improvements in health care delivery. Given the importance of understanding patient-reported outcomes during the management of chronic rhinosinusitis (CRS), PROMs will play an essential role in informing and tailoring the right intervention to the right patient. The objective of this systematic review was to identify and assess the quality of PROMs being used for adults with CRS. A systematic review of Ovid MEDLINE (R) (1947-May 2015), Embase, and the Cochrane databases was performed using the following key terms: ["chronic" AND "*sinusitis"] AND [PROM OR patient reported outcome measure* OR quality of life OR questionnaire OR survey OR valid* OR develop*]. An unlimited truncation strategy (placement of *) was used to capture all variations of terms used. The quality of each PROM was assessed and reported using standardized criteria from the COnsensus-based Standards for the selection of health status Measurement INstruments checklist. A total of 15 PROMs validated for use in adult patients with CRS were identified. Fourteen instruments were specific to adults with CRS, and one was a generic quality-of-life instrument (EuroQol five-dimensional questionnaire [EQ-5D]). There was significant variation in the quality of development and reporting of psychometric properties. Overall, the highest quality validated PROMs for adults with CRS were (1) the 22-item Sinonasal Outcome Test (19 points), (2) the Questionnaire of Olfactory Disorders (14 points), (3) the Sinusitis Control Test (14 points), and (4) the EQ-5D (13 points). Most of the PROMs were developed for research purposes such as determining changes in health-related quality of life or symptoms after an intervention as opposed to improving clinical decision making. Based on quality assessment, the 22-item Sinonasal Outcome Test, the Questionnaire of Olfactory Disorders, and the Sinusitis Control Test provided the highest quality CRS-specific PROMs, whereas the EQ-5D provided the highest quality generic quality-of-life instrument. Future CRS PROMs will need to incorporate clinical domains that assess common comorbid diseases along with patient values and preferences to improve clinical decision making.


Morbidity and mortality after colorectal procedures: comparison of data from the American College of Surgeons case log system and the ACS NSQIP.
Improving the quality of surgical care depends upon collection of robust data. The American College of Surgeons Case Log System enables surgeons to self-report patient risk factors and outcomes. In contrast, the American College of Surgeons National Surgical Quality Improvement Program (NSQIP) uses trained data abstractors to record similar data and uses a strict data collection methodology. The objective of this study was to assess bias in data entry for colorectal cases by comparing data in these 2 registries. One year of NSQIP (July 1, 2008 to June 30, 2009) and 7 years of Case Log (2003 to 2010) data were examined. Colorectal cases were identified by current procedural terminology code. The frequencies of comparably defined variables were compared, and mortality models were developed using logistic regression. Observed and expected mortality rates were compared. Rates of most risk factor and outcome variables were significantly higher in NSQIP than those in Case Log. NSQIP had a higher unadjusted mortality rate (4.46% versus 3.69%, p < 0.001); however, the adjusted odds of mortality was significantly higher in Case Log (odds ratio 1.32, p < 0.05). The Case Log model overpredicted mortality in NSQIP by 22%, whereas the NSQIP model underpredicted mortality in Case Log by 12%. Significant differences exist between risk factor and outcome data in NSQIP and Case Log for colorectal procedures. These differences demonstrate the need for standardized data collection methods, as is required by NSQIP, including use of standard definitions, adherence to a follow-up period for outcomes, and use of audits. These measures would improve the validity of using a self-reported database to evaluate and benchmark performance.


A survey of goiter morbidity in Ban Mae Toen, northwest Thailand.
A number of cases of goiter have been reported from Ban Mae Toen (BMT) northwestern Thailand, therefore we carried out this study to evaluate the prevalence and severity of goiter in BMT. We suspected fluoride toxicity as a factor in this phenomenon, therefore we used a WHO model using photographs to carry out a non-medical survey for teeth discoloration and limb deformity at the same time as the goiter survey. Every resident of BMT and a sampled population from two nearby control villages were surveyed in 2007. The overall prevalence of goiter among both sexes was 11%; 20% in women and 3% in men. No cases of goiter were recognised in the two control villages. Twenty-four percent of the BMT population and 5% of the sampled population of the control villages had lower limb deformities. Sixty-three percent of the BMT population had discoloration of teeth, especially among those > 35 years old. The control villages had significantly (p < 0.01) fewer cases of discoloration of teeth than the BMT population. The morbidity seen in Ban Mae Toen suggests the need for urgent enquiry into the etiology of this morbidity.


Oculomotor corollary discharge signaling is related to repetitive behavior in children with autism spectrum disorder.
Corollary discharge (CD) signals are "copies" of motor signals sent to sensory regions that allow animals to adjust sensory consequences of self-generated actions. Autism spectrum disorder (ASD) is characterized by sensory and motor deficits, which may be underpinned by altered CD signaling. We evaluated oculomotor CD using the blanking task, which measures the influence of saccades on visual perception, in 30 children with ASD and 35 typically developing (TD) children. Participants were instructed to make a saccade to a visual target. Upon saccade initiation, the presaccadic target disappeared and reappeared to the left or right of the original position. Participants indicated the direction of the jump. With intact CD, participants can make accurate perceptual judgements. Otherwise, participants may use saccade landing site as a proxy of the presaccadic target and use it to inform perception. We used multilevel modeling to examine the influence of saccade landing site on trans-saccadic perceptual judgements. We found that, compared with TD participants, children with ASD were more sensitive to target displacement and less reliant on saccade landing site when spatial uncertainty of the post-saccadic target was high. This pattern was driven by ASD participants with less severe restricted and repetitive behaviors. These results suggest a relationship between altered CD signaling and core ASD symptoms.


Magnetic resonance imaging appearance of cartilage repair in the knee.
Assessment of surgically repaired cartilage lesions with standardized cartilage sensitive magnetic resonance imaging was done to evaluate the integrity, morphologic features, and signal of the articular surface, thereby obtaining information about the natural history of these procedures in the knee. Magnetic resonance imaging also assessed the interface between the repaired and native cartilage, changes in the subchondral bone, and the appearance of cartilage over the opposite and adjacent (native) surfaces. One hundred eighty magnetic resonance imaging examinations were obtained in 112 patients who had cartilage-resurfacing procedures, including 86 microfractures and 35 autologous chondrocyte implantations, at a mean of 15 and 13 months after surgery, respectively. Autologous chondrocyte implantations showed consistently better fill of the defects at all times compared with microfracture. The graft hypertrophied in 63% of surgeries. The repair cartilage over the microfracture generally was depressed with respect to native cartilage. Propensity for bony overgrowth was most marked in the microfracture group, with loss of adjacent cartilage evident with progressive followup.


A tale of two studies of two disasters: comparing psychosocial responses to disaster among Oklahoma City bombing survivors and Hurricane Katrina evacuees.
An accumulation of disaster mental health research literature in the last few decades has contributed knowledge to direct disaster mental health interventions. However, no single set of principles can necessarily outline all anticipated mental health needs to be encountered in a particular disaster. To illustrate how different disaster scenarios may yield a divergence of mental health needs, this article compares mental health findings from two distinctly different studies of two very different populations affected by two very different disasters: directly exposed survivors the Oklahoma City bombing and sheltered evacuees from Hurricane Katrina. Research on the two disasters reviewed illustrates many facets and complexities of postdisaster mental health needs in different populations in different settings after different types of disasters. The major findings of the Oklahoma City bombing study related to posttraumatic stress disorder and the main findings of the Hurricane Katrina study involved need for treatment of preexisting chronic mental health and substance abuse problems. The disaster studies in this review diverged in type of disaster, affected populations, setting, and timing of the study, and these studies yielded a divergence of findings. One disaster mental health model clearly cannot adequately describe all postdisaster scenarios.


A multicenter, double-blinded validation study of methylation biomarkers for progression prediction in Barrett's esophagus.
Esophageal adenocarcinoma risk in Barrett's esophagus (BE) is increased 30- to 125-fold versus the general population. Among all BE patients, however, neoplastic progression occurs only once per 200 patient-years. Molecular biomarkers are therefore needed to risk-stratify patients for more efficient surveillance endoscopy and to improve the early detection of progression. We therefore performed a retrospective, multicenter, double-blinded validation study of eight BE progression prediction methylation biomarkers. Progression or nonprogression were determined at 2 years (tier 1) and 4 years (tier 2). Methylation was assayed in 145 nonprogressors and 50 progressors using real-time quantitative methylation-specific PCR. Progressors were significantly older than nonprogressors (70.6 versus 62.5 years; P < 0.001). We evaluated a linear combination of the eight markers, using coefficients from a multivariate logistic regression analysis. Areas under the ROC curve (AUC) were high in the 2-year, 4-year, and combined data models (0.843, 0.829, and 0.840; P < 0.001, <0.001, and <0.001, respectively). In addition, even after rigorous overfitting correction, the incremental AUCs contributed by panels based on the 8 markers plus age versus age alone were substantial (Delta-AUC = 0.152, 0.114, and 0.118, respectively) in all 3 models. A methylation biomarker-based panel to predict neoplastic progression in BE has potential clinical value in improving both the efficiency of surveillance endoscopy and the early detection of neoplasia.


Jump-Squat and Half-Squat Exercises: Selective Influences on Speed-Power Performance of Elite Rugby Sevens Players.
The aim of this study was to evaluate the relation between the maximum mean propulsive power (MPP) obtained in the loaded jump squat (JS) and half squat (HS) exercises and functional performances in vertical jumps, 40 m linear speed (VEL) and change-of-direction (COD) tests, using the median split technique. Twenty-two male rugby sevens players from the Brazilian National Olympic Team (Rio-2016) performed vertical jumping tests (squat and countermovement jumps [SJ and CMJ]), JS and HS exercises, COD speed test and sprinting velocity in 40 m, in this order. Based on the results of the MPP in the JS and HS exercises the participants were divided, using the median split, into four groups as follows: higher JS, lower JS, higher HS, and lower HS. Between-group differences in the functional tasks were detected via magnitude-based inferences. The athletes with higher MPP in the JS were capable of jumping higher and sprinting faster (including the COD speed test) than their weaker counterparts. This pattern was not observed in the HS exercise. To conclude, JS was shown to be more connected to sprinting, COD speed and jumping abilities than HS in elite rugby sevens players and should be preferred for assessing and possibly training elite athletes needing to improve speed-power related abilities.


High infiltration of CD68+ macrophages is associated with poor prognoses of head and neck squamous cell carcinoma patients and is influenced by human papillomavirus.
Incidence of human papillomavirus (HPV)-related head and neck squamous cell carcinomas (HNSCCs) has increased over the last few decades. The reaction of the host immune system to these tumors remains biologically complex. Here, we investigated CD68+ macrophage numbers, reporting the prognostic value in comparison to other risk factors. We also examined CD68+ macrophage infiltration during disease progression regarding the impact of HPV infection, and we studied the role of HPV16-E6/E7 oncoproteins in CD68+ macrophage recruitment. CD68+ macrophage numbers were evaluated in 10 cases of tumor-free peri-tumoral epithelia, 43 cases of low-grade dysplasia, 45 cases of high-grade dysplasia and 110 cases of carcinoma. Our in vivo model was developed in 80 C3H/HeN mice orthotopically injected with HPV16-E6, -E7 or -E6/E7-transfected SCC-VII cell lines. High CD68+ macrophage numbers in the intra-tumoral compartment were associated with shorter patient survival (recurrence-free survival: p = 0.001; overall survival: p = 0.01). Multivariate analyses reported that CD68+ macrophage infiltration and tumor stage were strong and independent prognostic factors of HNSCC. CD68+ macrophage numbers increased during HNSCC progression both in intra-epithelial (p < 0.001) and stromal compartments (p < 0.001). A higher density of CD68+ macrophages was observed in advanced stages (p = 0.004). Patients with transcriptionally active HPV infections had higher CD68+ macrophage density than did HPV-negative patients (p = 0.003). CD68+ macrophage infiltration was higher in HPV-E7+ and -E6/E7+ mouse tumors than in -E6+ tumors (p = 0.029 and p < 0.001). In conclusion, the extent of CD68+ macrophage infiltration is a significant prognostic factor for HNSCC patients. The recruitment of macrophages increases during disease progression and is influenced by the HPV virus.


The Effect of Organic Lithium Salts on Plasma 8-Hydroxy-2'-Deoxyguanosine in Bipolar Patients In Vitro.
An important step in the development of new drugs for the treatment of bipolar disorder (BD) is the study of the extent to which novel lithium salts whose anionic component has an antioxidant effect can reduce oxidative DNA damage in human blood plasma in vitro. We investigated the effects of lithium salts containing different organic anionic components (lithium carbonate (Li-CAR), pyruvate (Li-PYR), succinate (Li-SUC), fumarate (Li-FUM) and ascorbate (Li-ASC)) on levels of the oxidative damage product of DNA-8-hydroxy-2'-deoxyguanosine (8-OH-dG) in blood plasma after incubation of blood samples from healthy individuals (healthy group) and patients with bipolar disorder (BD-group) with these chemical compounds. Blood incubation was carried out in the presence of lithium salts (1.2 mM) for 1 hour at 37°C. Measurement of 8-OH-dG concentrations in blood plasma was carried out by enzyme immunoassay using a DNA Damage Competitive Elisa Kit (Thermo Fisher Scientific, USA). In samples without compounds (control), concentrations of 8-OH-dG in the BD-group did not differ from the group of healthy individuals. None of the tested compounds had a significant effect on 8-OH-dG in healthy individuals. In BD patients, Li-PYR significantly reduced levels of plasma 8-OH-dG, while other compounds did not have a noticeable effect. Lithium pyruvate reduces oxidative DNA damage in the blood of BD patients in vitro, demonstrating the potential of this compound to function not only as a mood stabilizer, but also as an antioxidant and cytoprotector.


Possible mechanisms of thyroid hormone disruption in mice by BDE 47, a major polybrominated diphenyl ether congener.
Polybrominated diphenyl ethers (PBDEs) are a class of polyhalogenated aromatic compounds commercially used as fire retardants in consumer products. These compounds have been shown to decrease thyroid hormone concentrations in rodents after acute exposures. This study examines the ability of 2,2',4,4'-tetrabromodiphenyl ether (BDE 47) to decrease circulating thyroid hormone concentrations and pairs this with BDE 47-induced effects on genes involved in thyroid hormone homeostasis. Female C57BL/6 mice (9 weeks old) were orally administered 3, 10, or 100 mg/kg/day of BDE 47 for 4 days. Animals were euthanized 24 h after the final dose (day 5) and liver, kidney, and serum were collected for analysis. BDE 47 caused a significant 43% decrease at 100 mg/kg/day in serum total thyroxine (T(4)) concentrations. There was no increase in hepatic T(4)-glucuronidation activity, but significant increases in hepatic Ugt1a1, Ugt1a7, and Ugt2b5 mRNA expression accompany significant decreases in T(4) concentrations at 100 mg/kg/day of BDE 47. Induction of PROD activity occurred at the lowest dose (3 mg/kg/day). Cyp2b10 mRNA expression also increased significantly at 10 and 100 mg/kg/day. These key findings show that BDE activates the nuclear receptor, CAR. Decreases in Mdr1a mRNA expression also occurred at the lowest dose administered (3 mg/kg/day BDE 47). BDE 47 exposure also decreased hepatic transthyretin and monocarboxylate transporter 8 (Mct8) mRNA expression, suggesting that while induction of UGTs may be partly responsible for T(4) decreases, other mechanisms are also involved. No effects were seen in the kidney. We conclude that changes in hepatic UGTs and transporters may be involved in decreases in circulating T(4) following BDE 47 exposure.


Central thalamic inactivation impairs the expression of action- and outcome-related responses of medial prefrontal cortex neurons in the rat.
The mediodorsal (MD) and adjacent intralaminar (IL) and midline nuclei provide the main thalamic input to the medial prefrontal cortex (mPFC) and are critical for associative learning and decision-making. MD neurons exhibit activity related to actions and outcomes that mirror responses of mPFC neurons in rats during dynamic delayed non-match to position (dDNMTP), a variation of DNMTP where start location is varied randomly within an open octagonal arena to avoid confounding behavioral events with spatial location. To test whether the thalamus affects the expression of these responses in mPFC, we inhibited the central thalamus unilaterally by microinjecting muscimol at doses and sites found to affect decision-making when applied bilaterally. Unilateral inactivation reduced normalized task-related responses in the ipsilateral mPFC without disrupting behavior needed to characterize event-related neuronal activity. Our results extend earlier findings that focused on delay-related activity by showing that central thalamic inactivation interferes with responses related to actions and outcomes that occur outside the period of memory delay. These findings are consistent with the broad effects of central thalamic lesions on behavioral measures of reinforcement-guided responding. Most (7/8) of the prefrontal response types affected by thalamic inactivation have also been observed in MD during dDNMTP. These results support the hypothesis that MD and IL act as transthalamic gates: monitoring prefrontal activity through corticothalamic inputs; integrating this information with signals from motivational and sensorimotor systems that converge in thalamus; and acting through thalamocortical projections to enhance expression of neuronal responses in the PFC that support adaptive goal-directed behavior.


Laparoscopic stoma formation with a prophylactic prosthetic mesh.
One year after stoma formation with an open technique, the rate of parastomal hernia is almost 50%. The herniation rate can be reduced to 10% with the use of a prophylactic mesh in a sublay position. For stomas formed with a laparoscopic technique, a surgical method with the use of prophylactic mesh should be sought. Patients with a sigmoidostomy created with a laparoscopic technique were provided with a prophylactic large-pore, low-weight mesh in a sublay position. Follow-up examination was carried out after at least 12 months. Between March 2003 and May 2007, a sigmoidostomy was created in 25 patients. The patients' mean age was 65 years (range 31-89), the mean body mass index was 26 (range 21-32) and 15 were female. One stoma necrosis and two minor wound infections occurred. Parastomal hernia was present in 3 of 20 patients (15%) available for follow-up examination after 11-31 months (mean 19). No fistulas or strictures had developed. No mesh infection was noted and no mesh was removed. In laparoscopic stoma formation, a prophylactic large-pore, low-weight mesh in a sublay position is an easy and safe procedure associated with a low rate of parastomal hernia.


Intracellular calcium transient of working human myocardium of seven patients transplanted for congestive heart failure.
The afterload dependence of the intracellular calcium transient in isolated working human myocardium was analyzed in both donor and recipient hearts of seven patients undergoing transplantation because of dilated cardiomyopathy. The intracellular calcium transient (recorded by the fura 2 ratio method), force development, and muscle shortening were simultaneously recorded in small (0.6 x 4.0-mm) electrically driven (60 beats per minute) trabeculas contracting at constant preload against varying afterloads. When the fibers contracted under isometric conditions, the intracellular calcium transients of normal and failing myocardium were similar. However, in dilated cardiomyopathy, stepwise afterload reduction and the concomitant increase in shortening amplitudes were associated with extraordinary alterations in the shape of the calcium transients; the amplitude rose, the time to peak was delayed, and at minimal afterloads, a long-lasting plateau was observed, and the diastolic decay was retarded. The calcium-time integral during shortening against passive resting force was 124 +/- 5% of the isometric control in normal myocardium and 172 +/- 12% in end-stage heart failure (P < .0001). We conclude that adequate interpretation of intracellular calcium transients requires simultaneous recordings of force and shortening. The extraordinary afterload dependence of the calcium transient in end-stage heart failure may be attributed to increased dissociation of calcium from the contractile proteins, a reduced calcium reuptake rate of the sarcoplasmic reticulum, or an increased calcium inflow due to altered permeabilities of the calcium channels during shortening. A potential role of mechanosensitive calcium channels has to be considered.


A Prospective Six Sigma Quality Improvement Trial to Optimize Universal Screening for Genetic Syndrome Among Patients With Young-Onset Colorectal Cancer.
Improving the quality of health care is a national priority, and providing patient-centered care is one of the 6 key areas for quality improvement. In the setting of patients with young-onset colorectal cancer (CRC), appropriate genetic workup and testing for potential underlying inherited CRC syndromes is fundamental to patient-centered care. Lynch syndrome (LS) is the most common of these inherited syndromes, and current recommendations from the NCCN and other professional societies advocate universal screening for LS among young patients with CRC. However, practical implementation of these guidelines often falls short. We conducted a prospective quality improvement intervention trial to optimize universal screening for LS in young (age <50 years) patients, involving 356 eligible patients during the 12-month preintervention period and 299 patients during the postintervention. Applying the Six Sigma conceptual framework, we demonstrated a significant increase in use of tumor-based molecular testing and subsequent confirmatory germline mutation testing for LS. This led to identification of more patients to be managed as having LS and of more first- and second-degree relatives to benefit from the testing results. This study demonstrated the successful application of a quality improvement conceptual framework for the universal adoption of molecular biomarker testing in patients with cancer, and for improving adherence to NCCN Clinical Practice Guidelines in Oncology for CRC Screening. As molecular and genetic testing is becoming increasingly common, we present a prototype study for improving the adoption of molecular studies and the provision of guideline-based patient-centered care.


P300 component in euthymic patients with bipolar disorder type I, bipolar disorder type II and healthy controls: a preliminary event-related potential study.
The aim of the present study was to investigate P300 event-related potential components in euthymic bipolar disorder type I (BDI) and bipolar disorder type II (BDII) patients and matched controls. A total of 10 BDI patients, 10 BDII patients and 10 healthy individuals were enrolled in the study. Event-related potential data were collected according to a standard auditory 'oddball' paradigm. A significant groups effect in both the peak amplitude (P<0.001) and the mean amplitude (P<0.001) was observed; post-hoc comparisons showed that the peak and mean amplitudes of BDI and BDII patients were significantly lower than the peak and mean amplitudes of the healthy controls. The neurophysiological patterns found in the present study might at least partially reflect the presence of a mild selective cognitive impairment in euthymic BDI and BDII patients. From a clinical point of view, these evidences support the potential role of cognitive interventions in the treatment of BD.


The relationship between foot arch measurements and walking parameters in children.
Walking mechanics are influenced by body morphology. Foot arch height is one aspect of body morphology central to walking. However, generalizations about the relationship between arch height and walking are limited due to previous methodologies used for measuring the arch and the populations that have been studied. To gain the knowledge needed to support healthy gait in children and adults, we need to understand this relationship in unimpaired, typically developing children and adults using dynamic measures. The purpose of the current study was to examine the relationship between arch height and gait in a sample of healthy children and adults using dynamic measures. Data were collected from 638 participants (n = 254 children and n = 384 adults) at the Museum of Science, Boston (MOS) and from 18 4- to 8-year-olds at the Motor Development and Motor Control Laboratories. Digital footprints were used to calculate two arch indices: the Chippaux-Smirak (CSI) and the Keimig Indices (KI). The height of the navicular bone was measured. Gait parameters were captured with a mechanized gait carpet at the MOS and three-dimensional motion analyses and in-ground force plates in the Motor Development and Motor Control Laboratories. Linear regression analyses on data from the MOS confirmed that as age increases, step length increases. With a linear mixed effect regression model, we found that individuals who took longer steps had higher arches as measured by the KI. However, this relationship was no longer significant when only adults were included in the model. A model restricted to children found that amongst this sample, those with higher CSI and higher KI values take longer relative step lengths. Data from the Motor Development and Motor Control Laboratories showed that both CSI and KI added to the prediction; children with lower anterior ground reaction forces had higher CSI and higher KI values. Arch height indices were correlated with navicular height. These results suggest that more than one measure of the arch may be needed elucidate the relationship between arch height and gait.


Genetic structure of Trypanosoma cruzi in Colombia revealed by a High-throughput Nuclear Multilocus Sequence Typing (nMLST) approach.
Chagas disease is a systemic pathology caused by Trypanosoma cruzi. This parasite reveals remarkable genetic variability, evinced in six Discrete Typing Units (DTUs) named from T. cruzi I to T. cruzi VI (TcI to TcVI). Recently newly identified genotypes have emerged such as TcBat in Brazil, Colombia and Panama associated to anthropogenic bats. The genotype with the broadest geographical distribution is TcI, which has recently been associated to severe cardiomyopathies in Argentina and Colombia. Therefore, new studies unraveling the genetic structure and natural history of this DTU must be pursued. We conducted a spatial and temporal analysis on 50 biological clones of T. cruzi I (TcI) isolated from humans with different clinical phenotypes, triatomine bugs and mammal reservoirs across three endemic regions for Chagas disease in Colombia. These clones were submitted to a nuclear Multilocus Sequence Typing (nMLST) analysis in order to elucidate its genetic diversity and clustering. After analyzing 13 nuclear housekeeping genes and obtaining a 5821 bp length alignment, we detected two robust genotypes within TcI henceforth named TcIDOM (associated to human infections) and a second cluster associated to peridomestic and sylvatic populations. Additionaly, we detected putative events of recombination and an intriguing lack of linkage disequilibrium. These findings reinforce the emergence of an enigmatic domestic T. cruzi genotype (TcIDOM), and demonstrates the high frequency of recombination at nuclear level across natural populations of T. cruzi. Therefore, the need to pursue studies focused on the diferential virulence profiles of TcI strains. The biological and epidemiological implications of these findings are herein discussed.


Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5' terminal nucleotide.
Argonaute (AGO) proteins recruit small RNAs to form the core of RNAi effector complexes. Arabidopsis encodes ten AGO proteins and a large network of small RNAs. How these small RNAs are sorted into specific AGO complexes remains largely unknown. We have cataloged small RNAs resident in four AGO complexes. We found that AGO2 and AGO4 preferentially recruit small RNAs with a 5' terminal adenosine, whereas AGO1 harbors microRNAs (miRNAs) that favor a 5' terminal uridine. AGO5 predominantly binds small RNAs that initiate with cytosine. Changing the 5' terminal nucleotide of an miRNA predictably redirected it into a different AGO complex and alters its biological activity. These results reveal a role for small RNA sequences in assorting among AGO complexes. This suggests that specialization of AGO complexes might involve remodeling the 5' end-binding pocket to accept certain small RNA sequences, perhaps explaining the evolutionary drive for miRNAs to initiate with uridine.


Three-dimensional spatiotemporal focusing of holographic patterns.
Two-photon excitation with temporally focused pulses can be combined with phase-modulation approaches, such as computer-generated holography and generalized phase contrast, to efficiently distribute light into two-dimensional, axially confined, user-defined shapes. Adding lens-phase modulations to 2D-phase holograms enables remote axial pattern displacement as well as simultaneous pattern generation in multiple distinct planes. However, the axial confinement linearly degrades with lateral shape area in previous reports where axially shifted holographic shapes were not temporally focused. Here we report an optical system using two spatial light modulators to independently control transverse- and axial-target light distribution. This approach enables simultaneous axial translation of single or multiple spatiotemporally focused patterns across the sample volume while achieving the axial confinement of temporal focusing. We use the system's capability to photoconvert tens of Kaede-expressing neurons with single-cell resolution in live zebrafish larvae.


Microscope-integrated intraoperative OCT with electrically tunable focus and heads-up display for imaging of ophthalmic surgical maneuvers.
We present novel optical and mechanical designs for a microscope-integrated intraoperative optical coherence tomography (iOCT) system with enhanced function and ergonomics for visualization of ophthalmic surgical maneuvers. Integration of an electrically tunable lens allows rapid focal plane adjustment and iOCT imaging of both anterior and posterior segment tissue microstructures while maintaining parfocality with the ophthalmic surgical microscope. We demonstrate novel visualization of instrument positions relative to tissue layers of interest as colormap overlays onto en face OCT data, which may provide integrative display of volumetric information during surgical maneuvers. Finally, we implement a heads-up display system to provide real-time feedback as proof-of-principle for iOCT-guided ophthalmic surgery.


Identification of Biomarkers for Differentiation of Hypervirulent Klebsiella pneumoniae from Classical K. pneumoniae.
A hypervirulent Klebsiella pneumoniae (hvKp) pathotype is undergoing global dissemination. In contrast to the usual health care-associated epidemiology of classical K. pneumoniae (cKp) infections, hvKp causes tissue-invasive infections in otherwise healthy individuals from the community, often involving multiple sites. An accurate test to identify hvKp strains is needed for improved patient care and epidemiologic studies. To fill this knowledge gap, clinical criteria or random blood isolates from North American and United Kingdom strain collections were used to assemble hvKp-rich (n = 85) and cKp-rich (n = 90) strain cohorts, respectively. The isolates were then assessed for multiple candidate biomarkers hypothesized to accurately differentiate the two cohorts. The genes peg-344, iroB, iucA, plasmid-borne rmpA gene ( prmpA), and prmpA2 all demonstrated >0.95 diagnostic accuracy for identifying strains in the hvKp-rich cohort. Next, to validate this epidemiological analysis, all strains were assessed experimentally in a murine sepsis model. peg-344, iroB, iucA, prmpA, and prmpA2 were all associated with a hazard ratio of >25 for severe illness or death, additionally supporting their utility for identifying hvKp strains. Quantitative siderophore production of ≥30 μg/ml also strongly predicted strains as members of the hvKp-rich cohort (accuracy, 0.96) and exhibited a hazard ratio of 31.7 for severe illness or death. The string test, a widely used marker for hvKp strains, performed less well, achieving an accuracy of only 0.90. Last, using the most accurate biomarkers to define hvKp, prevalence studies were performed on two Western strain collections. These data strongly support the utility of several laboratory markers for identifying hvKp strains with a high degree of accuracy.


Large herbivores promote habitat specialization and beta diversity of African savanna trees.
Edaphic variation in plant community composition is widespread, yet its underlying mechanisms are rarely understood and often assumed to be physiological. In East African savannas, Acacia tree species segregate sharply across soils of differing parent material: the ant-defended whistling thorn, A. drepanolobium (ACDR), is monodominant on cracking clay vertisols that are nutrient rich but physically stressful, whereas poorly defended species such as A. brevispica (ACBR) dominate on nutrient-poor but otherwise less-stressful sandy loams. Using a series of field experiments, we show that large-mammal herbivory interacts with soil properties to maintain this pattern. In the absence of large herbivores, transplanted saplings of both species established on both soil types. Browsers strongly suppressed survival and growth of ACDR saplings on sandy soil, where resource limitation constrained defensive investment. On clay soil, ACBR saplings established regardless of herbivory regime, but elephants prevented recruitment to maturity, apparently because trees could not tolerate the combination of biotic and abiotic stressors. Hence, each tree species was filtered out of one habitat by browsing in conjunction with different edaphic factors and at different ontogenetic stages. Browser abundance was greater on sandy soil, where trees were less defended, consistent with predicted feedbacks between plant community assembly and herbivore distributions. By exploring two inversely related axes of soil "quality" (abiotic stress and nutrient content), our study extends the range of mechanisms by which herbivores are known to promote edaphic specialization, illustrates how the high cost of a protection mutualism can constrain the realized niche of host trees, and shows that large-scale properties of savanna ecosystems are shaped by species interactions in cryptic ways that mimic simple abiotic determinism. These results suggest that ongoing declines in large-herbivore populations may relax spatial heterogeneity in plant assemblages and reduce the beta diversity of communities.


Low glucose sensitivity and polymodal chemosensing in neonatal rat adrenomedullary chromaffin cells.
Glucose is the primary metabolic fuel in mammalian fetuses, yet mammals are incapable of endogenous glucose production until several hours after birth. Thus, when the maternal supply of glucose ceases at birth there is a transient hypoglycemia that elicits a counterregulatory surge in circulating catecholamines. Because the innervation of adrenomedullary chromaffin cells (AMCs) is immature at birth, we hypothesized that neonatal AMCs act as direct glucosensors, a property that could complement their previously established roles as hypoxia and acid hypercapnia sensors. During perforated-patch, whole cell recordings, low glucose depolarized and/or excited a subpopulation of neonatal AMCs; in addition, aglycemia (0 mM glucose) caused inhibition of outward K(+) current, blunted by the simultaneous activation of glibenclamide-sensitive K(ATP) channels. Some cells were excited by each of the three metabolic stimuli, i.e., aglycemia, hypoxia (Po(2) ∼30 mmHg), and isohydric hypercapnia (10% CO(2); pH = 7.4). Using carbon fiber amperometry, aglycemia and hypoglycemia (3 mM glucose) induced robust catecholamine secretion that was sensitive to nickel (50 μM and 2 mM) and the L-type Ca(2+) channel blocker nifedipine (10 μM), suggesting involvement of both T-type and L-type voltage-gated Ca(2+) channels. Fura-2 measurements of intracellular Ca(2+) ([Ca(2+)] (i)) revealed that ∼42% of neonatal AMCs responded to aglycemia with a significant rise in [Ca(2+)] (i). Approximately 40% of these cells responded to hypoxia, whereas ∼25% cells responded to both aglycemia and hypoxia. These data suggest that together with hypoxia and acid hypercapnia, low glucose is another important metabolic stimulus that contributes to the vital asphyxia-induced catecholamine surge from AMCs at birth.


Adjuvant pharmacotherapy in the management of elderly patients with pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) is the fourth or fifth leading cause of death from cancer in Western industrialized countries. Surgical resection is the only chance of cure, but only 15-20 % of cases are potentially resectable at presentation, and despite complete resection, the overall prognosis remains relatively poor. Adjuvant therapy has modestly improved cure rates. The majority of patients with pancreatic cancer are over the age of 65 years. But this age group is underrepresented within clinical trials, and it is unknown whether older patients achieve similar results to younger ones in terms of survival and treatment tolerance. In addition, there are no clinical trials dedicated to the elderly. Retrospective studies coming from the non-resectable setting provide some understanding on outcomes in older patients with PDAC. To date, we can reasonably argue that selected elderly patients with PDAC can benefit from curative surgery and postoperative chemotherapy as do their younger counterparts, without a significant increase in morbidity and mortality. Gemcitabine should be preferred to 5-fluorouracil on the basis of a better risk-benefit balance.


Switch recombination breakpoints are strictly correlated with DNA recognition motifs for immunoglobulin S gamma 3 DNA-binding proteins.
The deletion looping out model of switch (S) recombination predicts that the intervening DNA between switch regions will be excised as a circle. Circular excision products of immunoglobulin switch recombination have been recently isolated from lipopolysaccharide (LPS)-stimulated spleen cells. The recombination breakpoints in these large circles were found to fall within switch regions. Since switch recombination is clearly focused on switch regions, we hypothesized that some DNA-binding protein factor might be involved in specifically recognizing and facilitating the alignment of switch regions before recombination. Two DNA-binding proteins that specifically interact with two discrete regions of the S gamma 3 tandem repeat have been identified in crude and partially purified nuclear extracts derived from LPS- and dextran sulfate (DxS)-activated splenic B cells. The first factor has been found indistinguishable from NF-kappa B by mobility shift assays, methylation interference, competition binding studies, and supershift analysis using an antiserum specific for the p50 component. The second appears to be composed of two closely traveling mobilities that do not separate upon partial purification. This second complex is unique and specific for S gamma 3 by methylation interference assays and competition-binding analysis. The sites at which recombination occurs in the S gamma 3 switch region have been analyzed and found to strictly correlate with the binding sites of the S gamma 3 switch binding proteins.


Laboratory costs of a hospital-based blood transfusion service in Malawi.
Despite policies advocating centralised transfusion services based on voluntary donors, the hospital-based replacement donor system is widespread in sub-Saharan Africa. To evaluate the cost of all laboratory resources needed to provide a unit of safe blood in rural Malawi using the family replacement donor system Full economic costs of all laboratory tests used to screen potential donors and to perform cross-matching were documented in a prospective, observational study in Ntcheu district hospital laboratory. 1729 potential donors were screened and 11,008 tests were performed to ensure that 1104 units of safe blood were available for transfusion. The annual cost of all transfusion-related tests (in 2005 USdollars) was USdollars 17,976, equivalent to USdollars 16.28 per unit of transfusion-ready blood. Transfusion-related tests used 53% of the laboratory's total annual expenditure of USdollars 33,608. This is the first study to provide prospective economic costs of all laboratory tests associated with the family replacement donor system in a district hospital in Africa. Results show that despite potential economies of scale, a unit of blood from the centralised system costs about three times as much as one from the hospital-based "replacement" system. Factors affecting these relative costs are complex but are in part due to the cost of donor recruitment in centralised systems. In the replacement system the cost of donor recruitment is entirely borne by families of patients needing a blood transfusion.


X-ray structure of a voltage-dependent K+ channel.
Voltage-dependent K+ channels are members of the family of voltage-dependent cation (K+, Na+ and Ca2+) channels that open and allow ion conduction in response to changes in cell membrane voltage. This form of gating underlies the generation of nerve and muscle action potentials, among other processes. Here we present the structure of KvAP, a voltage-dependent K+ channel from Aeropyrum pernix. We have determined a crystal structure of the full-length channel at a resolution of 3.2 A, and of the isolated voltage-sensor domain at 1.9 A, both in complex with monoclonal Fab fragments. The channel contains a central ion-conduction pore surrounded by voltage sensors, which form what we call 'voltage-sensor paddles'-hydrophobic, cationic, helix-turn-helix structures on the channel's outer perimeter. Flexible hinges suggest that the voltage-sensor paddles move in response to membrane voltage changes, carrying their positive charge across the membrane.


Sustaining cerebral perfusion in intracranial atherosclerotic stenosis: The roles of antegrade residual flow and leptomeningeal collateral flow.
We aimed to investigate the roles of antegrade residual flow and leptomeningeal collateral flow in sustaining cerebral perfusion distal to an intracranial atherosclerotic stenosis (ICAS). Patients with apparently normal cerebral perfusion distal to a symptomatic middle cerebral artery (MCA)-M1 stenosis were enrolled. Computational fluid dynamics models were built based on CT angiography to obtain a translesional pressure ratio (PR) to gauge the residual antegrade flow. Leptomeningeal collaterals (LMCs) were scaled on CT angiography. Cerebral perfusion metrics were obtained in CT perfusion maps. Among 83 patients, linear regression analyses revealed that both translesional PR and LMC scale were independently associated with relative ipsilesional mean transit time (rMTT). Subgroup analyses showed that ipsilesional rMTT was significantly associated with translesional PR (p < 0.001) rather than LMC scale in those with a moderate (50-69%) MCA stenosis, which, however, was only significantly associated with LMC scale (p = 0.051) in those with a severe (70-99%) stenosis. Antegrade residual flow and leptomeningeal collateral flow have complementary effects in sustaining cerebral perfusion distal to an ICAS, while cerebral perfusion may rely more on the collateral circulation in those with a severe stenosis.


Analysis of genetic variation in two human thyroxine-binding plasma proteins by immunodetection after isoelectric focusing.
An immunological method for the detection of two thyroxine-binding plasma proteins, thyroxine-binding globulin (TBG) and thyroxine-binding prealbumin (TBPA), following polyacrylamide gel isoelectric focusing is described. Both proteins show complex electrophoretic patterns, attributable to post-translational glycosylation, which are affected by unfavourable conditions of sample storage. No genetic variants of TBG or TBPA were encountered in a study of N. European individuals (n = 50). The X-linked TBG polymorphism, previously detected using a radiolabelled ligand binding technique in Pacific Islanders, was, however, demonstrable by our new immunological method in a previously untested population (n = 139) from the island of Maewo in Vanuatu.


Sputum matrix metalloproteinase-12 in patients with chronic obstructive pulmonary disease and asthma: relationship to disease severity.
Matrix metalloproteinase (MMP)-12 has been implicated in the pathogenesis of both chronic obstructive pulmonary disease (COPD) and asthma. The influence of disease severity on sputum MMP-12 concentrations and activity is not known. We sought to examine the relationship between disease severity assessed by means of lung function and computed tomography (CT) and induced sputum MMP-12 concentrations and activity in patients with asthma and COPD. In 208 subjects (109 asthmatic patients, smokers and never smokers, mild, moderate, and severe; 53 patients with COPD, smokers and exsmokers, mild, moderate, and severe; and 46 healthy control subjects, smokers and never smokers), we measured induced sputum MMP-12 concentrations (ELISA) and enzyme activity (fluorescence resonance energy transfer), sputum cell MMP12 mRNA expression (quantitative PCR [qPCR]), diffusing capacity for carbon monoxide (Dlco), and CT assessment of emphysema (percentage of low-attenuation areas at less -950 Hounsfield units). Sputum MMP-12 concentrations are greater in patients with COPD and smokers with asthma than in healthy nonsmokers (P = .003 and P = .035, respectively) but similar to those seen in healthy smokers. In patients with COPD, disease severity, when measured by means of CT-assessed emphysema, but not by means of spirometry or Dlco values, is directly associated with sputum MMP-12 concentrations and activity. In the asthma groups there is no significant association between disease severity and sputum MMP-12 concentrations or activity. Sputum MMP-12 concentrations and activity in patients with COPD are directly associated with the extent of emphysema measured by means of CT. This finding supports a role for MMP-12 in the pathogenesis of COPD and might suggest that blocking MMP-12 activity in patients with COPD could prevent the further development of emphysema.


Presynaptic silencing is an endogenous neuroprotectant during excitotoxic insults.
Glutamate release is a root cause of acute and delayed neuronal damage in response to hypoxic/ischemic insults. Nevertheless, therapeutics that target the postsynaptic compartment have been disappointing clinically. Here we explored whether presynaptic silencing (muting) of glutamatergic terminals is sufficient to reduce excitotoxic damage resulting from hypoxia and oxygen/glucose deprivation. Our evidence suggests that strong depolarization, previously shown to mute glutamate synapses, protects neurons by a presynaptic mechanism that is sensitive to inhibition of the proteasome. Postsynaptic Ca2+ rises in response to glutamate application and toxicity in response to exogenous glutamate treatment were unaffected by depolarization preconditioning. These features strongly suggest that reduced glutamate release explains preconditioning protection. We addressed whether hypoxic depolarization itself induces presynaptic silencing, thereby participating in the damage threshold for hypoxic insult. Indeed, we found that the hypoxic insult increased the percentage of mute glutamate synapses in a proteasome-dependent manner. Furthermore, proteasome inhibition exacerbated neuronal loss to mild hypoxia and prevented hypoxia-induced muting. In total our results suggest that presynaptic silencing is an endogenous neuroprotective mechanism that could be exploited to reduce damage from insults involving excess synaptic glutamate release.


The Chiropractic Hospital-based Interventions Research Outcomes (CHIRO) study: a randomized controlled trial on the effectiveness of clinical practice guidelines in the medical and chiropractic management of patients with acute mechanical low back pain.
Evidence-based clinical practice guidelines (CPGs) for the management of patients with acute mechanical low back pain (AM-LBP) have been defined on an international scale. Multicenter clinical trials have demonstrated that most AM-LBP patients do not receive CPG-based treatments. To date, the value of implementing full and exclusively CPG-based treatment remains unclear. To determine if full CPGs-based study care (SC) results in greater improvement in functional outcomes than family physician-directed usual care (UC) in the treatment of AM-LBP. A two-arm, parallel design, prospective, randomized controlled clinical trial using blinded outcome assessment. Treatment was administered in a hospital-based spine program outpatient clinic. Inclusion criteria included patients aged 19 to 59 years with Quebec Task Force Categories 1 and 2 AM-LBP of 2 to 4 weeks' duration. Exclusion criteria included "red flag" conditions and comorbidities contraindicating chiropractic spinal manipulative therapy (CSMT). improvement from baseline in Roland-Morris Disability Questionnaire (RDQ) scores at 16 weeks. improvements in RDQ scores at 8 and 24 weeks; and in Short Form-36 (SF-36) bodily pain (BP) and physical functioning (PF) scale scores at 8, 16, and 24 weeks. Patients were assessed by a spine physician, then randomized to SC (reassurance and avoidance of passive treatments, acetaminophen, 4 weeks of lumbar CSMT, and return to work within 8 weeks), or family physician-directed UC, the components of which were recorded. Ninety-two patients were recruited, with 36 SC and 35 UC patients completing all follow-up visits. Baseline prognostic variables were evenly distributed between groups. The primary outcome, the unadjusted mean improvement in RDQ scores, was significantly greater in the SC group than in the UC group (p=.003). Regarding unadjusted mean changes in secondary outcomes, improvements in RDQ scores were also greater in the SC group at other time points, particularly at 24 weeks (p=.004). Similarly, improvements in SF-36 PF scores favored the SC group at all time points; however, these differences were not statistically significant. Improvements in SF-36 BP scores were similar between groups. In repeated-measures analyses, global adjusted mean improvement was significantly greater in the SC group in terms of RDQ (p=.0002), nearly significantly greater in terms of SF-36 PF (p=.08), but similar between groups in terms of SF-36 BP (p=.27). This is the first reported randomized controlled trial comparing full CPG-based treatment, including spinal manipulative therapy administered by chiropractors, to family physician-directed UC in the treatment of patients with AM-LBP. Compared to family physician-directed UC, full CPG-based treatment including CSMT is associated with significantly greater improvement in condition-specific functioning.


Expansion of myeloid immune suppressor Gr+CD11b+ cells in tumor-bearing host directly promotes tumor angiogenesis.
We demonstrate a novel tumor-promoting role of myeloid immune suppressor Gr+CD11b+ cells, which are evident in cancer patients and tumor-bearing animals. These cells constitute approximately 5% of total cells in tumors. Tumors coinjected with Gr+CD11b+ cells exhibited increased vascular density, vascular maturation, and decreased necrosis. These immune cells produce high levels of MMP9. Deletion of MMP9 in these cells completely abolishes their tumor-promoting ability. Gr+CD11b+ cells were also found to directly incorporate into tumor endothelium. Consistent with this observation, Gr+CD11b+ cells acquire endothelial cell (EC) properties in tumor microenvironment and proangiogenic culture conditions. Our data provide evidence that Gr+CD11b+ cells of immune origin induced by tumors directly contribute to tumor growth and vascularization by producing MMP9 and differentiating into ECs.


Alanine Substitution Inactivates Cross-Reacting Epitopes in Dengue Virus Recombinant Envelope Proteins.
The expansion of the habitat of mosquitoes belonging to the Aedes genus puts nearly half of the world's population at risk of contracting dengue fever, and a significant fraction will develop its serious hemorrhagic complication, which can be fatal if not diagnosed properly and treated in a timely fashion. Although several diagnostic methods have been approved for dengue diagnostics, their applicability is limited in rural areas of developing countries by sample preparation costs and methodological requirements, as well as cross-reactivity among the different serotypes of the Dengue virus and other flavivirus, such as the Zika virus. For these reasons, it is necessary to generate more specific antigens to improve serological methods that could be cheaper and used in field operations. Here, we describe a strategy for the inactivation of cross-reacting epitopes on the surface of the Dengue virus envelope protein through the synthetic generation of recombinant peptide sequences, where key amino acid residues from Dengue virus serotype 1 (DENV-1) and 2 (DENV-2) are substituted by alanine residues. The proteins thus generated are recognized by 88% of sera from Dengue NS1+ patients and show improved serotype specificity because they do not react with the antibodies present in seroconverted, PCR-serotyped DEN-4 infected patients.


Peristalsis in the junction region of the Drosophila larval midgut is modulated by DH31 expressing enteroendocrine cells.
The underlying cellular and molecular mechanisms that coordinate the physiological processes in digestion are complex, cryptic, and involve the integration of multiple cellular and organ systems. In all intestines, peristaltic action of the gut moves food through the various stages of digestion from the anterior end towards the posterior, with the rate of flow dependent on signals, both intrinsic and extrinsic to the gut itself. We have identified an enteroendocrine cell type that regulates gut motility in the Drosophila melanogaster larval midgut. These cells are located at the junction of the anterior and the acidic portions of the midgut and are a group of enteroendocrine cells that express the peptide hormone Diuretic Hormone 31 in this region of the gut. Using cell ablation and ectopic activation via expression of the Chlamydomonas reinhardtii blue light-activated channelopsin, we demonstrate that these enteroendocrine cells are both necessary and sufficient for the peristalsis in the junction region of the midgut and require the Diuretic Hormone 31 to affect normal peristalsis in this region. Within the same junction region of the midgut, we have also identified morphological features suggesting that this region acts as a valve that regulates the transit of food from the anterior midgut into the acidic portion of the gut. We have characterized and described a set of enteroendocrine cells called the Midgut Junction DH31 expressing cells that are required for peristaltic movement in the junction region between the anterior portion and acidic region of the larval midgut of Drosophila melanogaster. We have shown that the Midgut Junction DH31 expressing cells are necessary and sufficient for motility and that the peptide hormone DH31 is required for peristalsis in the junction region of the midgut. The Drosophila model system will allow for a further dissection of the digestion process and provide a better understanding of the mechanisms that regulate digestion in all organisms.


Volcanic activity and climatic changes.
Radiocarbon dates of volcanic activity suggest variations that appear to be related to climatic changes. Historical eruption records also show variations on the scale of years to centuries. These records can be combined with simple climatic models to estimate the impact of various volcanic activity levels. From this analysis it appears that climatic prediction in the range of 2 years to many decades requires broad-scale volcanic activity prediction. Statistical analysis of the volcanic record suggests that some predictability is possible.