Any versus long-term prescribing of high risk medications in older people using 2012 Beers Criteria: results from three cross-sectional samples of primary care records for 2003/4, 2007/8 and 2011/12.
High risk medications are commonly prescribed to older US patients. Currently, less is known about high risk medication prescribing in other Western Countries, including the UK. We measured trends and correlates of high risk medication prescribing in a subset of the older UK population (community/institutionalized) to inform harm minimization efforts. Three cross-sectional samples from primary care electronic clinical records (UK Clinical Practice Research Datalink, CPRD) in fiscal years 2003/04, 2007/08 and 2011/12 were taken. This yielded a sample of 13,900 people aged 65 years or over from 504 UK general practices. High risk medications were defined by 2012 Beers Criteria adapted for the UK. Using descriptive statistical methods and regression modelling, prevalence of 'any' (drugs prescribed at least once per year) and 'long-term' (drugs prescribed all quarters of year) high risk medication prescribing and correlates were determined. While polypharmacy rates have risen sharply, high risk medication prevalence has remained stable across a decade. A third of older (65+) people are exposed to high risk medications, but only half of the total prevalence was long-term (any = 38.4 % [95 % CI: 36.3, 40.5]; long-term = 17.4 % [15.9, 19.9] in 2011/12). Long-term but not any high risk medication exposure was associated with older ages (85 years or over). Women and people with higher polypharmacy burden were at greater risk of exposure; lower socio-economic status was not associated. Ten drugs/drug classes accounted for most of high risk medication prescribing in 2011/12. High risk medication prescribing has not increased over time against a background of increasing polypharmacy in the UK. Half of patients receiving high risk medications do so for less than a year. Reducing or optimising the use of a limited number of drugs could dramatically reduce high risk medications in older people. Further research is needed to investigate why the oldest old and women are at greater risk. Interventions to reduce high risk medications may need to target shorter and long-term use separately.


Folic acid administration produces an antidepressant-like effect in mice: evidence for the involvement of the serotonergic and noradrenergic systems.
Clinical studies have shown that folic acid plays a role in the pathophysiology of depression. However, very few studies have investigated its effect in behavioral models of depression. Hence, this study tested its effect in the forced swimming test (FST) and the tail suspension test (TST), two models predictive of antidepressant activity, in mice. Folic acid administered by oral route (p.o.) produced a reduction in the immobility time in the FST (50-100mg/kg) and in the TST (10-50mg/kg). The administration of folic acid by i.c.v. route also reduced the immobility time in the FST (10nmol/site) and in the TST (1-10nmol/site). Both folic acid administered by oral and i.c.v. route produced no psychostimulant effect, which indicates that its antidepressant-like effect is specific. Pretreatment of mice with p-chlorophenylalanine methyl ester (PCPA; 100mg/kg, i.p., an inhibitor of serotonin (5-HT) synthesis, for 4 consecutive days), ketanserin (5mg/kg, i.p., a 5-HT(2A/2C) receptor antagonist), prazosin (1mg/kg, i.p., an alpha(1)-adrenoceptor antagonist) or yohimbine (1mg/kg, i.p., an alpha(2)-adrenoceptor antagonist) prevented the anti-immobility effect of folic acid (50mg/kg, p.o.) in the FST. Moreover, the pretreatment of mice with WAY100635 (0.1mg/kg, s.c., a selective 5-HT(1A) receptor antagonist) blocked the decrease in immobility time in the FST elicited by folic acid (50mg/kg, p.o.), but produced a synergistic effect with a subeffective dose of folic acid (10mg/kg, p.o.). In addition, a subeffective dose of folic acid (10mg/kg, p.o.) produced a synergistic antidepressant-like effect with fluoxetine (10mg/kg, p.o.) in the FST. Overall, the results firstly indicate that folic acid produced an antidepressant-like effect in FST and in TST and that this effect appears to be mediated by an interaction with the serotonergic (5-HT(1A) and 5-HT(2A/2C) receptors) and noradrenergic (alpha(1)- and alpha(2)-adrenoceptors) systems.


Vaccines for the prevention of seasonal influenza in patients with diabetes: systematic review and meta-analysis.
Patients with diabetes are at increased risk of severe influenza disease; influenza vaccination for these patients is therefore recommended by the World Health Organization and several National Immunization Technical Advisory Groups. However, no systematic review has evaluated the effects of influenza vaccines for patients with diabetes. We conducted a systematic review and meta-analysis by searching Medline, Embase, Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov from inception until November 2014. We included all types of studies reporting on the efficacy, effectiveness, and/or safety of influenza vaccination in patients with type 1 and type 2 diabetes of all ages. We used the Newcastle-Ottawa scale to assess risk of bias in observational studies. Residual confounding was addressed by comparing estimates of vaccine effectiveness (VE) during influenza seasons to those obtained during off-seasons. Quality of the evidence for each outcome was assessed using the GRADE methodology. Following review of 1,444 articles, 11 observational studies with a total of 170,924 participants were included. In diabetic patients of working-age (18-64 years), influenza vaccination prevented all-cause hospitalization with a pooled VE of 58% (95% CI, 6-81%) and hospitalization due to influenza or pneumonia (VE 43%; 95% CI, 28-54%), whereas no effects on all-cause mortality and influenza-like illness (ILI) were observed. In the elderly (65+), influenza vaccination prevented all-cause mortality (VE 38%; 95% CI, 32-43%), all-cause hospitalization (VE 23%; 95% CI, 1-40%), hospitalization due to influenza or pneumonia (VE 45%; 95% CI, 34-53%), and ILI (VE 13%; 95% CI, 10-16%). However, significant off-season estimates for several outcomes indicated residual confounding, particularly in elderly patients. Quality of the evidence was low to very low for all outcomes. Laboratory-confirmed influenza infections were not reported. Due to strong residual confounding in most of the identified studies, the available evidence is insufficient to determine the magnitude of benefit that diabetic people derive from seasonal influenza vaccination. Adequately powered randomized controlled trials or quasi-experimental studies using laboratory-confirmed influenza-specific outcomes are urgently needed.


The functional impact of HIV-associated neuropsychological impairment in Spanish-speaking adults: a pilot study.
Among English-speaking adults, HIV-associated neuropsychological (NP) impairments have been associated with problems in everyday functioning, including ability to function at work and drive an automobile. Latinos account for a disproportionate number of HIV/AIDS cases nationwide, and a significant segment of this population is primarily Spanish speaking. We have previously developed an assessment that evaluates English-speakers on a variety of instrumental activities of daily living. In this pilot study, we used Spanish-language translations of our functional battery to investigate the cultural relevance of such measures, and to explore relationships between NP status and ability to perform important everyday tasks in HIV-infected Spanish-speakers. Sixteen HIV-infected monolingual Spanish-speaking adults received comprehensive, Spanish language NP testing and functional assessments included the following domains: Medication Management, Cooking, Finances, Shopping, and Restaurant Scenario. Results revealed that most of the functional tasks appeared culturally relevant and appropriate with minor modifications. NP-impaired participants were significantly more functionally impaired compared to NP-normals (88% vs. 13%, p <.01). Performances on the functional assessment and the NP battery were also related to indicators of real world functioning, including employment status and quality of life. These results, though preliminary, suggest that Spanish language functional assessments are potentially valid tools for detecting everyday functioning deficits associated with NP impairments in HIV-infected Spanish-speakers.


Systemic Staphylococcus aureus infection mediated by Candida albicans hyphal invasion of mucosal tissue.
Candida albicans and Staphylococcus aureus are often co-isolated in cases of biofilm-associated infections. C. albicans can cause systemic disease through morphological switch from the rounded yeast to the invasive hyphal form. Alternatively, systemic S. aureus infections arise from seeding through breaks in host epithelial layers although many patients have no documented portal of entry. We describe a novel strategy by which S. aureus is able to invade host tissue and disseminate via adherence to the invasive hyphal elements of Candida albicans. In vitro and ex vivo findings demonstrate a specific binding of the staphylococci to the candida hyphal elements. The C. albicans cell wall adhesin Als3p binds to multiple staphylococcal adhesins. Furthermore, Als3p is required for C. albicans to transport S. aureus into the tissue and cause a disseminated infection in an oral co-colonization model. These findings suggest that C. albicans can facilitate the invasion of S. aureus across mucosal barriers, leading to systemic infection in co-colonized patients.


Inference of type-specific HPV transmissibility, progression and clearance rates: a mathematical modelling approach.
Quantifying rates governing the clearance of Human Papillomavirus (HPV) and its progression to clinical disease, together with viral transmissibility and the duration of naturally-acquired immunity, is essential in estimating the impact of vaccination programmes and screening or testing regimes. However, the complex natural history of HPV makes this difficult. We infer the viral transmissibility, rate of waning natural immunity and rates of progression and clearance of infection of 13 high-risk and 2 non-oncogenic HPV types, making use of a number of rich datasets from Sweden. Estimates of viral transmissibility, clearance of initial infection and waning immunity were derived in a Bayesian framework by fitting a susceptible-infectious-recovered-susceptible (SIRS) transmission model to age- and type-specific HPV prevalence data from both a cross-sectional study and a randomised controlled trial (RCT) of primary HPV screening. The models fitted well, but over-estimated the prevalence of four high-risk types with respect to the data. Three of these types (HPV-33, -35 and -58) are among the most closely related phylogenetically to the most prevalent HPV-16. The fourth (HPV-45) is the most closely related to HPV-18; the second most prevalent type. We suggest that this may be an indicator of cross-immunity. Rates of progression and clearance of clinical lesions were additionally estimated from longitudinal data gathered as part of the same RCT. Our estimates of progression and clearance rates are consistent with the findings of survival analysis studies and we extend the literature by estimating progression and clearance rates for non-16 and non-18 high-risk types. We anticipate that such type-specific estimates will be useful in the parameterisation of further models and in developing our understanding of HPV natural history.


Binocular rivalry is not chaotic.
Time series of the durations each eye was dominant during binocular rivalry were obtained psychophysically. The oscillations showed an adaptation effect with mean and standard deviations of rivalry dominance durations increasing as a square root function of time over the course of a trial. The data were corrected for this non-stationarity. Dominance durations had a log-normal probability distribution and the autocorrelation function revealed no short term correlations in the time series. In an attempt to distinguish whether the variability of durations was due to a deterministic, low-dimensional chaotic attractor or to a stochastic process, the data were subjected to two tests. The first was calculation of correlation dimensions and the second was nonlinear forecasting of the time series. Both tests included comparisons with randomized 'surrogate data' as controls. In neither case was there a large difference between test results for actual data and surrogate data. We conclude that chaos is not a major factor underlying variability in binocular rivalry.


Airway epithelial barrier dysfunction in the pathogenesis and prognosis of respiratory tract diseases in childhood and adulthood.
The lungs are in direct contact with the environment through the tubular structure that constitutes the airway. Starting from the nasal orifice, the airway is exposed to foreign particles including infectious agents, allergens, and other substances that can damage the airways. Therefore, the airway must have a functional epithelial barrier both in the upper and lower airways to protect against these threats. As with the skin, it is likely that the pathogenesis of respiratory diseases is a consequence of epithelial barrier defects in these airways. The characteristics of this system, starting from the beginning of life and extending into maturing and aging, determine the prognosis of respiratory diseases. In this article, we discuss the pathogenesis, clinical phenotype, and prognosis of respiratory diseases from newborns to adulthood in the context of epithelial barrier function and dysfunction.


Hypoxia-induced miR-210 modulates the inflammatory response and fibrosis upon acute ischemia.
Hypoxia-induced miR-210 is a crucial component of the tissue response to ischemia, stimulating angiogenesis and improving tissue regeneration. Previous analysis of miR-210 impact on the transcriptome in a mouse model of hindlimb ischemia showed that miR-210 regulated not only vascular regeneration functions, but also inflammation. To investigate this event, doxycycline-inducible miR-210 transgenic mice (Tg-210) and anti-miR-210 LNA-oligonucleotides were used. It was found that global miR-210 expression decreased inflammatory cells density and macrophages accumulation in the ischemic tissue. To dissect the underpinning cell mechanisms, Tg-210 mice were used in bone marrow (BM) transplantation experiments and chimeric mice underwent hindlimb ischemia. MiR-210 overexpression in the ischemic tissue was sufficient to increase capillary density and tissue repair, and to reduce inflammation in the presence of Wt-BM infiltrating cells. Conversely, when Tg-210-BM cells migrated in a Wt ischemic tissue, dysfunctional angiogenesis, inflammation, and impaired tissue repair, accompanied by fibrosis were observed. The fibrotic regions were positive for α-SMA, Vimentin, and Collagen V fibrotic markers and for phospho-Smad3, highlighting the activation of TGF-β1 pathway. Identification of Tg-210 cells by in situ hybridization showed that BM-derived cells contributed directly to fibrotic areas, where macrophages co-expressing fibrotic markers were observed. Cell cultures of Tg-210 BM-derived macrophages exhibited a pro-fibrotic phenotype and were enriched with myofibroblast-like cells, which expressed canonical fibrosis markers. Interestingly, inhibitors of TGF-β type-1-receptor completely abrogated this pro-fibrotic phenotype. In conclusion, a context-dependent regulation by miR-210 of the inflammatory response was identified. miR-210 expression in infiltrating macrophages is associated to improved angiogenesis and tissue repair when the ischemic recipient tissue also expresses high levels of miR-210. Conversely, when infiltrating an ischemic tissue with mismatched miR-210 levels, macrophages expressing high miR-210 levels display a pro-fibrotic phenotype, leading to impaired tissue repair, fibrosis, and dysfunctional angiogenesis.


Morphology and function of Neandertal and modern human ear ossicles.
The diminutive middle ear ossicles (malleus, incus, stapes) housed in the tympanic cavity of the temporal bone play an important role in audition. The few known ossicles of Neandertals are distinctly different from those of anatomically modern humans (AMHs), despite the close relationship between both human species. Although not mutually exclusive, these differences may affect hearing capacity or could reflect covariation with the surrounding temporal bone. Until now, detailed comparisons were hampered by the small sample of Neandertal ossicles and the unavailability of methods combining analyses of ossicles with surrounding structures. Here, we present an analysis of the largest sample of Neandertal ossicles to date, including many previously unknown specimens, covering a wide geographic and temporal range. Microcomputed tomography scans and 3D geometric morphometrics were used to quantify shape and functional properties of the ossicles and the tympanic cavity and make comparisons with recent and extinct AMHs as well as African apes. We find striking morphological differences between ossicles of AMHs and Neandertals. Ossicles of both Neandertals and AMHs appear derived compared with the inferred ancestral morphology, albeit in different ways. Brain size increase evolved separately in AMHs and Neandertals, leading to differences in the tympanic cavity and, consequently, the shape and spatial configuration of the ossicles. Despite these different evolutionary trajectories, functional properties of the middle ear of AMHs and Neandertals are largely similar. The relevance of these functionally equivalent solutions is likely to conserve a similar auditory sensitivity level inherited from their last common ancestor.


The Rd8 mutation of the Crb1 gene is present in vendor lines of C57BL/6N mice and embryonic stem cells, and confounds ocular induced mutant phenotypes.
We noted an unexpected inheritance pattern of lesions in several strains of gene-manipulated mice with ocular phenotypes. The lesions, which appeared at various stages of backcross to C57BL/6, bore resemblance to the rd8 retinal degeneration phenotype. We set out to examine the prevalence of this mutation in induced mutant mouse lines, vendor C57BL/6 mice and in widely used embryonic stem cells. Ocular lesions were evaluated by fundus examination and histopathology. Detection of the rd8 mutation at the genetic level was performed by PCR with appropriate primers. Data were confirmed by DNA sequencing in selected cases. Analysis of several induced mutant mouse lines with ocular disease phenotypes revealed that the disease was associated 100% with the presence of the rd8 mutation in the Crb1 gene rather than with the gene of interest. DNA analysis of C57BL/6 mice from common commercial vendors demonstrated the presence of the rd8 mutation in homozygous form in all C57BL/6N substrains, but not in the C57BL/6J substrain. A series of commercially available embryonic stem cells of C57BL/6N origin and C57BL/6N mouse lines used to generate ES cells also contained the rd8 mutation. Affected mice displayed ocular lesions typical of rd8, which were detectable by funduscopy and histopathology as early as 6 weeks of age. These findings identify the presence of the rd8 mutation in the C57BL/6N mouse substrain used widely to produce transgenic and knockout mice. The results have grave implications for the vision research community who develop mouse lines to study eye disease, as presence of rd8 can produce significant disease phenotypes unrelated to the gene or genes of interest. It is suggested that researchers screen for rd8 if their mouse lines were generated on the C57BL/6N background, bear resemblance to the rd8 phenotype, or are of indeterminate origin.


Tonic current through GABAA receptors and hyperpolarization-activated cyclic nucleotide-gated channels modulate resonance properties of rat subicular pyramidal neurons.
The subiculum, considered to be the output structure of the hippocampus, modulates information flow from the hippocampus to various cortical and sub-cortical areas such as the nucleus accumbens, lateral septal region, thalamus, nucleus gelatinosus, medial nucleus and mammillary nuclei. Tonic inhibitory current plays an important role in neuronal physiology and pathophysiology by modulating the electrophysiological properties of neurons. While the alterations of various electrical properties due to tonic inhibition have been studied in neurons from different regions, its influence on intrinsic subthreshold resonance in pyramidal excitatory neurons expressing hyperpolarization-activated cyclic nucleotide-gated (HCN) channels is not known. Using pharmacological agents, we show the involvement of α5βγ GABAA receptors in the picrotoxin-sensitive tonic current in subicular pyramidal neurons. We further investigated the contribution of tonic conductance in regulating subthreshold electrophysiological properties using current clamp and dynamic clamp experiments. We demonstrate that tonic GABAergic inhibition can actively modulate subthreshold properties, including resonance due to HCN channels, which can potentially alter the response dynamics of subicular pyramidal neurons in an oscillating neuronal network.


Percutaneous alcohol sclerotherapy of simple hepatic cysts. Results from a multicentre survey in Italy.
The increased use of Ultrasonography (US) has led to increased detection of simple hepatic cysts. For symptomatic cysts treatment is necessary. Until some years ago surgery was the only therapy. We have treated a large number of patients with Percutaneous Alcohol Sclerotherapy (PAS) and evaluated retrospectively the efficacy of this approach. Data on 21 patients with symptomatic simple hepatic cysts were reviewed retrospectively. Cysts had a mean diameter of 9 cm (range: 7-15 cm). PAS was always performed under local anesthesia and US guidance. 25% of the volume was replaced with 95% ethanol and then completely aspirated after 20-30 minutes. No complications or deaths occurred. In all patients symptoms disappeared after treatment. In 15 out of 21 cases there was no evidence of residual cyst on US, computed tomography (CT) or magnetic resonance (MRI). In 6 patients with shorter follow-up, cysts showed a mean reduction in diameter of 50%. The mean follow-up was 18 months (range 6-60 months). We conclude that PAS is easy with low risk for the patients and with good long-term results; it should therefore become the procedure of choice for simple hepatic cysts.


Understanding the limits to generalizability of experimental evolutionary models.
Given the difficulty of testing evolutionary and ecological theory in situ, in vitro model systems are attractive alternatives; however, can we appraise whether an experimental result is particular to the in vitro model, and, if so, characterize the systems likely to behave differently and understand why? Here we examine these issues using the relationship between phenotypic diversity and resource input in the T7-Escherichia coli co-evolving system as a case history. We establish a mathematical model of this interaction, framed as one instance of a super-class of host-parasite co-evolutionary models, and show that it captures experimental results. By tuning this model, we then ask how diversity as a function of resource input could behave for alternative co-evolving partners (for example, E. coli with lambda bacteriophages). In contrast to populations lacking bacteriophages, variation in diversity with differences in resources is always found for co-evolving populations, supporting the geographic mosaic theory of co-evolution. The form of this variation is not, however, universal. Details of infectivity are pivotal: in T7-E. coli with a modified gene-for-gene interaction, diversity is low at high resource input, whereas, for matching-allele interactions, maximal diversity is found at high resource input. A combination of in vitro systems and appropriately configured mathematical models is an effective means to isolate results particular to the in vitro system, to characterize systems likely to behave differently and to understand the biology underpinning those alternatives.


Tuning the mobility coupling of quaternized polyvinylpyridine and anionic phospholipids in supported lipid bilayers.
Binding of biomacromolecules to anionic lipids in the plasma membrane is a common motif in many cell signaling pathways. Previous work has shown that macromolecules with cationic sequences can form nanodomains with sequestered anionic lipids, which alters the lateral distribution and mobility of the membrane lipids. Such sequestration is believed to result from the formation of a lipid-macromolecule complex. To date, however, the molecular structure and dynamics of the lipid-polymer interface are poorly understood. We have investigated the behavior of polycationic quaternized polyvinylpyridine (QPVP) on supported lipid bilayers doped with phosphatidylserine (PS) or phosphatidylinositol phosphate (PIP) lipids using time-resolved fluorescence microscopy, including pulsed interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS). PIE-FCCS is a dual-color fluorescence spectroscopy that translates fluctuations in fluorescence signal into a measurement of diffusion and colocalization. By labeling the polymer and lipids, we investigated the adsorption-induced translational mobility of lipids and systematically studied the influence of lipid charge density and solution ionic strength. Our results show that alteration of anionic lipid lateral mobility is dependent on the net charge of the lipid headgroup and is modulated by the ionic strength of the solution, indicating that electrostatic interactions drive the decrease in lateral mobility of anionic lipids by adsorbed QPVP. At physiological salt concentration we observe that the lipid lateral mobility is weakly influenced by QPVP and that there is no evidence of stable lipid-polymer complexes.


SELDI-TOF MS profiling of serum for detection of the progression of chronic hepatitis C to hepatocellular carcinoma.
Proteomic profiling of serum is an emerging technique to identify new biomarkers indicative of disease severity and progression. The objective of our study was to assess the use of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) to identify multiple serum protein biomarkers for detection of liver disease progression to hepatocellular carcinoma (HCC). A cohort of 170 serum samples obtained from subjects in the United States with no liver disease (n = 39), liver diseases not associated with cirrhosis (n = 36), cirrhosis (n = 38), or HCC (n = 57) were applied to metal affinity protein chips for protein profiling by SELDI-TOF MS. Across the four test groups, 38 differentially expressed proteins were used to generate multiple decision classification trees to distinguish the known disease states. Analysis of a subset of samples with only hepatitis C virus (HCV)-related disease was emphasized. The serum protein profiles of control patients were readily distinguished from each HCV-associated disease state. Two-way comparisons of chronic hepatitis C, HCV cirrhosis, or HCV-HCC versus healthy had a sensitivity/specificity range of 74% to 95%. For distinguishing chronic HCV from HCV-HCC, a sensitivity of 61% and a specificity of 76% were obtained. However, when the values of known serum markers alpha fetoprotein, des-gamma carboxyprothrombin, and GP73 were combined with the SELDI peak values, the sensitivity and specifity improved to 75% and 92%, respectively. In conclusion, SELDI-TOF MS serum profiling is able to distinguish HCC from liver disease before cirrhosis as well as cirrhosis, especially in patients with HCV infection compared with other etiologies.


Potential Efficiency of Riparian Vegetated Buffer Strips in Intercepting Soluble Compounds in the Presence of Subsurface Preferential Flows.
Buffer strips have been widely recognized as to promote infiltration, deposition and sorption of contaminants for protecting surface water against agricultural contamination. However, such strips do not intercept all contaminants, particularly soluble ones. Although preferential flow (PF) has been suggested as one factor among several decreasing the efficiency of buffer strips, the mechanisms involved are not well understood. This project examines buffer strip efficiency at intercepting solutes when subsurface PF occurs. Two soluble sorbed tracers, FD&C Blue #1 and rhodamine WT, were applied on an agricultural sandy loam soil to evaluate the ability of a naturally vegetated buffer strip to intercept soluble contaminants. Rhodamine was applied about 15 m from the creek, while the Blue was applied 15 m to 165 m from the creek. Tracer concentration was measured over a two-year period in both the creek and the buffer strip through soil and water samples. Although the tracers traveled via different pathways, they both quickly moved toward the creek, passing beneath the buffer strip through the soil matrix. Our results demonstrate that the risk of water contamination by soluble contaminants is high in such systems, even when a well-vegetated buffer strip is used. The design of buffer strips should be modified to account for underground bypass, either by using plants that have deep, fine roots that do not favour PF or by adding a filter extending deep underground that can be regularly changed.


Isolation and Characterization of Agrobacterium Strains from Soil: A Laboratory Capstone Experience.
In this investigation, the students' goal was to isolate and characterize Agrobacterium strains from soil. Following selection and enrichment on 1A-t medium, putative Agrobacterium isolates were characterized by Gram stain reaction and biochemical tests. Isolates were further evaluated using polymerase chain reaction (PCR) with different primer sets designed to amplify specific regions of bacterial deoxyribonucleic acid (DNA). Primer sets included AGRH to identify isolates that were members of the Rhizobiaceae, BIOVAR1 primers to identify members of Agrobacterium biovar group I, and a third set, VIRG, to determine presence of virG (only present in pathogenic Agrobacterium strains). During the investigation, students applied previously learned techniques including serial dilution, use of selective/differential media, staining protocols, biochemical analysis, molecular analysis via PCR, and electrophoresis. Students also gained practical experience using photo documentation to record data for an eventual mock journal publication of the capstone laboratory experience. Pre- and post-evaluation of class content knowledge related to the techniques, protocols, and learning objectives of these laboratories revealed significant learning gains in the content areas of Agrobacterium-plant interactions (p ≤ 0.001) and molecular biology (p ≤ 0.01). The capstone journal assignment served as the assessment tool to evaluate mastery and application of laboratory technique, the ability to accurately collect and evaluate data, and critical thinking skills associated with experimental troubleshooting and extrapolation. Analysis of journal reports following the capstone experience showed significant improvement in assignment scores (p ≤ 0.0001) and attainment of capstone experience learning outcomes.


Poor trail making test performance is directly associated with altered dual task prioritization in the elderly--baseline results from the TREND study.
Deterioration of executive functions in the elderly has been associated with impairments in walking performance. This may be caused by limited cognitive flexibility and working memory, but could also be caused by altered prioritization of simultaneously performed tasks. To disentangle these options we investigated the associations between Trail Making Test performance--which specifically measures cognitive flexibility and working memory--and dual task costs, a measure of prioritization. Out of the TREND study (Tuebinger evaluation of Risk factors for Early detection of Neurodegenerative Disorders), 686 neurodegeneratively healthy, non-demented elderly aged 50 to 80 years were classified according to their Trail Making Test performance (delta TMT; TMT-B minus TMT-A). The subjects performed 20 m walks with habitual and maximum speed. Dual tasking performance was tested with walking at maximum speed, in combination with checking boxes on a clipboard, and subtracting serial 7 s at maximum speeds. As expected, the poor TMT group performed worse when subtracting serial 7 s under single and dual task conditions, and they walked more slowly when simultaneously subtracting serial 7 s, compared to the good TMT performers. In the walking when subtracting serial 7 s condition but not in the other 3 conditions, dual task costs were higher in the poor TMT performers (median 20%; range -6 to 58%) compared to the good performers (17%; -16 to 43%; p<0.001). To the contrary, the proportion of the poor TMT performance group that made calculation errors under the dual tasking situation was lower than under the single task situation, but higher in the good TMT performance group (poor performers, -1.6%; good performers, +3%; p = 0.035). Under most challenging conditions, the elderly with poor TMT performance prioritize the cognitive task at the expense of walking velocity. This indicates that poor cognitive flexibility and working memory are directly associated with altered prioritization.


Revision rhinoplasty.
Revision rhinoplasty requires sound judgment by both patient and surgeon. Detailed preoperative assessment facilitates proper patient selection. If the patient is deemed a favorable revision rhinoplasty candidate, both patient and surgeon must agree on the goals for and risks inherent in revision rhinoplasty. Thoughtfulness regarding the degree of aesthetic and functional correction desired by the patient and the corresponding intraoperative techniques required to achieve this correction must be contemplated prior to committing to revision rhinoplasty. Furthermore, the surgeon must feel confident that the surgical demands requested by the patient may be met and satisfaction achieved. A precise preoperative surgical plan minimizes intraoperative misadventures. Options for grafting materials should be discussed and agreed upon by both patient and surgeon preoperatively. Both patient and surgeon commit to long-term postoperative follow-up and the patience necessary for the eventual outcome to be realized.


Optimization of vincristine-topotecan combination--paving the way for improved chemotherapy regimens by nanoliposomes.
There is an opportunity to improve the therapeutic potential of a combination of two drugs using nanoliposomes. The combination of topotecan (TPT) and vincristine (VCR) was selected. The ratio-dependent synergy between these two drugs was evaluated, in an attempt to improve the therapeutic efficacy of this combination in vivo. The interaction between the drugs was evaluated in tissue culture by the median-effect analysis. Certain ratios of combined drugs were synergistic, whereas, others were antagonistic, implying that the most efficacious combinations should be at a specific fixed drug ratio. For in vivo evaluation, nanoliposomes co-remotely loaded simultaneously with both drugs by transmembrane ammonium sulfate gradient were developed. VCR and TPT were successfully co-encapsulated at therapeutically relevant levels in the same nanoliposome (LipoViTo). The nanoliposomes controlled the drugs' "biofate" and maintained a fixed drug ratio in vivo, allowing one to compare the therapeutic efficacy of various predefined drug ratios. Pharmacokinetics and biodistribution studies showed that LipoViTo delivers the two drugs simultaneously to the tumors, where they are released at a predefined ratio. LipoViTo was more efficacious than the free drugs and liposomes with one agent, singly or in combination, in two tumor models in mice. LipoViTo co-loaded with both drugs corresponding to their maximal tolerated dose (MTD) ratio resulted in the best therapeutic efficacy. To summarize: liposomal co-encapsulation of anticancer drug combinations can profoundly influence therapeutic outcomes. Drug combinations can be optimized preclinically through pharmacokinetic control by remote loading into nanoliposomes.


Nitric oxide as an initiator of brain lesions during the development of Alzheimer disease.
Nitric oxide (NO) is an important regulatory molecule for the host defense that plays a fundamental role in the cardiovascular, immune, and nervous systems. NO is synthesized through the conversion of L-arginine to L-citrulline by the enzyme NO synthase (NOS), which is found in three isoforms classified as neuronal (nNOS), inducible (iNOS), and endothelial (eNOS). Recent evidence supports the theory that this bioactive molecule has an influential role in the disruption of normal brain and vascular homeostasis, a condition known to elucidate chronic hypoperfusion which ultimately causes the development of brain lesions and the pathology that typify Alzheimer disease (AD). In addition, vascular NO activity appears to be a major contributor to this pathology before any overexpression of NOS isoforms is observed in the neuron, glia, and microglia of the brain tree, where the overexpression the NOS isoforms causes the formation of a large amount of NO. We hypothesize that since an imbalance between the NOS isoforms and endothelin-1 (ET-1), a human gene that encodes for blood vessel constriction, can cause antioxidant system insufficiency; by using pharmacological intervention with NO donors and/or NO suppressors, the brain lesions and the downstream progression of brain pathology and dementia in AD should be delayed or minimized.


A call to action: The controversy of and rationale for competency-based medical education.
Although medical education has enjoyed many successes over the last century, there is a recognition that health care is too often unsafe and of poor quality. Errors in diagnosis and treatment, communication breakdowns, poor care coordination, inappropriate use of tests and procedures, and dysfunctional collaboration harm patients and families around the world. These issues reflect on our current model of medical education and raise the question: Are physicians being adequately prepared for twenty-first century practice? Multiple reports have concluded the answer is "no." Concurrent with this concern is an increasing interest in competency-based medical education (CBME) as an approach to help reform medical education. The principles of CBME are grounded in providing better and safer care. As interest in CBME has increased, so have criticisms of the movement. This article summarizes and addresses objections and challenges related to CBME. These can provide valuable feedback to improve CBME implementation and avoid pitfalls. We strongly believe medical education reform should not be reduced to an "either/or" approach, but should blend theories and approaches to suit the needs and resources of the populations served. The incorporation of milestones and entrustable professional activities within existing competency frameworks speaks to the dynamic evolution of CBME, which should not be viewed as a fixed doctrine, but rather as a set of evolving concepts, principles, tools, and approaches that can enable important reforms in medical education that, in turn, enable the best outcomes for patients.


Simulation of mouse and rat spermatogenesis to inform genotoxicity testing using OECD test guideline 488.
The Organisation for Economic Co-operation and Development Test Guideline (TG) 488 for the transgenic rodent (TGR) mutation assay recommends two sampling times for assessing germ cell mutagenicity following the required 28-day exposure period: 28 + > 49 days for mouse sperm and 28 + >70 days for rat sperm from the cauda epididymis, or three days (i.e., 28 + 3d) for germ cells from seminiferous tubules (hereafter, tubule germ cells) plus caudal sperm for mouse and rat. Although the latter protocol is commonly used for mutagenicity testing in somatic tissues, it has several shortcomings for germ cell testing because it provides limited exposure of the proliferating phase of spermatogenesis when mutations are fixed in the transgene. Indeed, analysis of sperm at 28 + 3d has generated negative results with established germ cell mutagens, while the analysis of tubule germ cells has generated both positive and either negative or equivocal results. The Germ Cell workgroup of the Genetic Toxicology Technical Committee of the Health and Environmental Sciences Institute modelled mouse and rat spermatogenesis to better define the exposure history of the cell population collected from seminiferous tubules. The modelling showed that mouse tubule germ cells at 28 + 3d receive, as a whole, 42% of the total exposure during the proliferating phase. This percentage increases to 99% at 28 + 28d and reaches 100% at 28 + 30d. In the rat, these percentages are 22% and 80% at 28 + 3d and 28 + 28d, reaching 100% at 28 + 44d. These results show that analysis of tubule germ cells at 28 + 28d may be an effective protocol for assessing germ cell mutagenicity in mice and rats using TG 488. Since TG 488 recommends the 28 + 28d protocol for slow dividing somatic tissues, this appears to be a better compromise than 28 + 3d when slow dividing somatic tissues or germ cells are the critical tissues of interest.


Antibiotics for asymptomatic bacteriuria in pregnancy.
Asymptomatic bacteriuria occurs in 2% to 10% of pregnancies and, if not treated, up to 30% of mothers will develop acute pyelonephritis. Asymptomatic bacteriuria has been associated with low birthweight and preterm birth. To assess the effect of antibiotic treatment for asymptomatic bacteriuria on the development of pyelonephritis and the risk of low birthweight and preterm birth. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (19 March 2015) and reference lists of retrieved studies. Randomized trials comparing antibiotic treatment with placebo or no treatment in pregnant women with asymptomatic bacteriuria found on antenatal screening. Two review authors independently assessed trials for inclusion and risk of bias, extracted data and checked them for accuracy. Fourteen studies, involving almost 2000 women, were included. Antibiotic treatment compared with placebo or no treatment reduced the incidence of pyelonephritis (average risk ratio (RR) 0.23, 95% confidence interval (CI) 0.13 to 0.41; 11 studies, 1932 women; very low quality evidence). Antibiotic treatment was also associated with a reduction in the incidence of low birthweight babies (average RR 0.64, 95% CI 0.45 to 0.93; six studies, 1437 babies; low quality evidence) and preterm birth (RR 0.27, 95% CI 0.11 to 0.62; two studies, 242 women; low quality evidence). A reduction in persistent bacteriuria at the time of delivery was seen (average RR 0.30, 95% CI 0.18 to 0.53; four studies; 596 women). There were very limited data on which to estimate the effect of antibiotics on other infant outcomes and maternal adverse effects were rarely described.Overall, all 14 studies were assessed as being at high or unclear risk of bias. While many studies lacked an adequate description of methods and the risk of bias could only be assessed as unclear, in almost all studies there was at least one domain where the risk of bias was judged as high. The three primary outcomes were assessed with GRADE software and given a quality rating. Evidence for pyelonephritis, preterm birth and birthweight less than 2500 g was assessed as of low or very low quality. While antibiotic treatment is effective in reducing the risk of pyelonephritis in pregnancy, the estimate of the effect is very uncertain because of the very low quality of the evidence. The reduction in low birthweight and preterm birth with antibiotic treatment is consistent with theories about the role of infection in adverse pregnancy outcomes, but this association should be interpreted with caution given the very poor quality of the included studies.


SU-E-I-31: Evaluating Brain Imaging Material (BIM) as a Brain Tissue Surrogate for Use in Neuro-Endovascular Imaged Guided Intervention (EIGI) Research.
A brain tissue surrogate material was needed to fill the anatomical cavity of a skull to create a phantom for use in simulated Neuro-EIGI procedures. To enable diagnostic and interventional procedure simulation, the BIM must fit into and be congruous with the interior surface of the skull, be reusable, and allow the implantation of vascular phantoms. The material must reasonably reproduce the automatic technique parameter selections observed during Neuro-EIGI procedures. We formulated a putty- like material to be used as the BIM. Its x-ray attenuation properties were evaluated by comparison of the fluoroscopic and radiographic technique parameters automatically selected for a BIM-filled skull on a Toshiba Infinix angiographic C-arm unit to those of a solid anthropomorphic head phantom at various projection angles. The same comparison was made between the skull phantom without BIM in the cavity and the anthropomorphic head phantom. The BIM linear attenuation coefficient was calculated and compared to that of PMMA, a common tissue analog plastic. The BIM keeps its shape, is moldable and reusable, and is congruent to the skull's interior surfaces. It allows for insertion and interchange of various custom vascular phantoms at proper anatomic locations. Addition of the BIM to the skull cavity improves the matching of the automatically selected parameters to those of the anthropomorphic phantom by an average of 96.3% for mAs and by 4.2% for kVp in fluoroscopy mode and by 88.6% and 9.0%, respectively, in DSA mode. The BIM's experimental and theoretical linear attenuation coefficient for the RQA5 spectrum differed from PMMA's by about 30%. Despite the difference in attenuation coefficients between the PMMA and BIM, the BIM is a good surrogate material for Neuro-EIGI research as shown by its properties of congruity, reusability, and device implantation, along with the demonstrated improvement of automatically selected technique parameters. Supported in part by: NIH Grants R01-EB008425, R01-EB002873, and an equipment grant from Toshiba Medical Systems Corp.


Six-week physical rehabilitation protocol for anterior shoulder dislocation in athletes.
Anterior shoulder dislocations are common in young athletes. The mechanism for the first or primary shoulder dislocation may involve a collision or a fall typically with the arm in an abducted and externally rotated position. The aim of this study was to design a physical rehabilitation program using the elastic band and resistive exercise to improve joint strength and range of motion in individuals diagnosed with a first-time shoulder dislocation. Twelve physically active males with a first-time acute shoulder dislocation were asked to volunteer. Participants began a physical rehabilitation program 2 weeks after the shoulder dislocation, which was confirmed by a referring physician. The rehabilitation program was 6 weeks in duration and required the participants to engage in progressive resistive loads/duration using elastic bands and weights 5 days per week. Pretest and posttest measures included shoulder strength and range of motion. All outcome measures were compared between the injured and uninjured shoulder, which served as the control condition in this study. There were statistically significant differences between the injured and uninjured shoulder for measures of strength and range of motion during pretests (P<0.01) but not post-tests (P<0.53). Finally, there were no differences between shoulders in regards to the volume measure suggesting that any changes in muscle atrophy or swelling were not detected. The physical rehabilitation program proposed in this study was effective at improving strength and range of motion in the injured shoulder as evidenced by the similarity in posttest values between the injured and uninjured shoulder.


A Relative Absence: Exploring Professional Experiences of Funerals Without Mourners.
When someone dies, it is usual for relatives to gather at a funeral to embody a collective act of eulogy for the deceased and stand against the finality of death. When someone who lived alone dies alone at home, it is not always possible to identify anyone to attend a funeral. In such cases, funeral professionals are required to perform the appropriate social rites in the absence of the confirmatory power of a society. Drawing on interviews with funeral professionals and ethnographic observations of funerals without mourners, we explore how professionals understand their roles in performing social rites against death when there is no one to participate in them. We consider the impact of attempting to make good a death generally perceived as bad, and we examine the significance of funerals as a social rite when the deceased is assumed to have forgone social relationships during their lifetime.


Physiologically-based pharmacokinetic modelling for the reduction of animal use in the discovery of novel pharmaceuticals.
The challenges of physiologically-based pharmacokinetic (PBPK) modelling and approaches to replacing the use of animals, in order to determine drug pharmacokinetics, are discussed. Reference is made to the limitations of in vivo animal studies in drug discovery. In particular, the ways in which animal studies contribute to drug attrition during the post-preclinical phase of testing are considered.


Use of lablab (Lablab purpureus (L.) Sweet) for bio-control by native arthropods and its effect on yield of pumpkins.
Silverleaf whitefly (SLW, Bemisia tabaci MEAM1) and aphids are sap-sucking insects, which pose a serious threat to Australian cucurbit crops and the horticulture industry. Traditional chemical control for these insect pests is becoming less effective, and there is a need to search for alternative or supplementary methods. This study aimed to manipulate the habitat of pumpkin crops in a tropical setting (Queensland, Australia), by growing pumpkins (var. Japanese pumpkin) alone and between lablab (Lablab purpureus L. Sweet). It was hypothesized that the presence of lablab will increase the populations of natural enemies, and through their control of insect pests such as SLW and aphids, will affect pumpkin yield. The population of arthropods (natural enemies and pests of pumpkin), with a focus on SLW and aphids, were sampled weekly on both lablab and pumpkin crop for a total of 21 weeks. Results showed that lablab hosted more enemies of SLW per plant than pumpkin in either treatment. In addition, adult SLW numbers were significantly higher in the pumpkin-only crop compared with the pumpkin grown between lablab, while pumpkin in the mixed plantings had significantly more ladybirds and lacewing larvae (P < 0.05). While there was no significant difference in the average fruit weight between treatments, the total weight (kg) and number of marketable pumpkins per hectare was greater (P < 0.05) for the pumpkin/lablab treatment than the pumpkin-only treatment. This study shows that growing lablab alongside a pumpkin crop may enhance natural enemies of SLW and could significantly increase the yield.


Presynaptic kainate receptors regulate spinal sensory transmission.
Small diameter dorsal root ganglion (DRG) neurons, which include cells that transmit nociceptive information into the spinal cord, are known to express functional kainate receptors. It is well established that exposure to kainate will depolarize C-fiber afferents arising from these cells. Although the role of kainate receptors on sensory afferents is unknown, it has been hypothesized that presynaptic kainate receptors may regulate glutamate release in the spinal cord. Here we show that kainate, applied at low micromolar concentrations in the presence of the AMPA-selective antagonist (RS)-4-(4-aminophenyl)-1, 2-dihydro-1-methyl-2-propyl-carbamoyl-6,7-methylenedioxyphthalazine++ +, suppressed spontaneous NMDA receptor-mediated EPSCs in cultures of spinal dorsal horn neurons. In addition, kainate suppressed EPSCs in dorsal horn neurons evoked by stimulation of synaptically coupled DRG cells in DRG-dorsal horn neuron cocultures. Interestingly, although the glutamate receptor subunit 5-selective kainate receptor agonist (RS)-2-alpha-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA) (2 micrometer) was able to suppress DRG-dorsal horn synaptic transmission to a similar extent as kainate (10 micrometer), it had no effect on excitatory transmission between dorsal horn neurons. Agonist applications revealed a striking difference between kainate receptors expressed by DRG and dorsal horn neurons. Whereas DRG cell kainate receptors were sensitive to both kainate and ATPA, most dorsal horn neurons responded only to kainate. Finally, in recordings from dorsal horn neurons in spinal slices, kainate and ATPA were able to suppress NMDA and AMPA receptor-mediated EPSCs evoked by dorsal root fiber stimulation. Together, these data suggest that kainate receptor agonists, acting at a presynaptic locus, can reduce glutamate release from primary afferent sensory synapses.


Relationship between foveal cone structure and clinical measures of visual function in patients with inherited retinal degenerations.
To study the relationship between cone spacing and density and clinical measures of visual function near the fovea. High-resolution images of the photoreceptor mosaic were obtained with adaptive optics scanning laser ophthalmoscopy from 26 patients with inherited retinal degenerations. Cone spacing measures were made close to or at the foveal center (mean [SD] eccentricity, 0.02 [0.03] degree; maximum eccentricity, 0.13 degree) and were converted to Z-scores, fraction of cones, and percentage-of-cones-below-average compared with normal values for each location (based on 37 age-similar visually normal eyes). Z-scores and percentage of cones below average were compared with best-corrected visual acuity (VA) and foveal sensitivity. Visual acuity was significantly correlated with cone spacing (Spearman rank correlation ρ = -0.60, P = 0.003) and was preserved (≥ 80 letters), despite cone density measures that were 52% below normal. Foveal sensitivity showed significant correlation with cone spacing (ρ = -0.47, P = 0.017) and remained normal (≥ 35 decibels), despite density measures that were approximately 52% to 62% below normal. Cone density was reduced by up to 62% below normal at or near the fovea in eyes with VA and sensitivity that remained within normal limits. Despite a significant correlation with foveal cone spacing, VA and sensitivity are insensitive indicators of the integrity of the foveal cone mosaic. Direct, objective measures of cone structure may be more sensitive indicators of disease severity than VA or foveal sensitivity in eyes with inherited retinal degenerations.


Modulation of endochondral ossification by MEK inhibitors PD0325901 and AZD6244 (Selumetinib).
MEK inhibitors (MEKi) PD0325901 and AZD6244 (Selumetinib) are drugs currently under clinical investigation for cancer treatment, however the Ras-MAPK pathway is also an important mediator of normal bone cell differentiation and function. In this study we examined the effects of these compounds on endochondral processes using both in vitro and in vivo models. Treatment with PD0325901 or AZD6244 significantly increased Runx2 and Alkaline phosphate gene expression in calvarial osteoblasts and decreased TRAP+ cells in induced osteoclast cultures. To test the effects of these drugs on bone healing, C57/Bl6 mice underwent a closed tibial fracture and were treated with PD0325901 or AZD6244 at 10mg/kg/day. Animals were culled at day 10 and at day 21 post-fracture for analysis of the fracture callus and the femoral growth plate in the contralateral leg. MEKi treatment markedly increased cartilage volume in the soft callus at day 10 post-fracture (+60% PD0325901, +20% AZD6244) and continued treatment led to a delay in cartilage remodeling. At the growth plate, we observed an increase in the height of the hypertrophic zone relative to the proliferative zone of +78% in PD0325901 treated mice. Osteoclast surface was significantly decreased both at the terminal end of the growth plate and within the fracture calluses of MEKi treated animals. The mechanistic effects of MEKi on genes encoding cartilage matrix proteins and catabolic enzymes were examined in articular chondrocyte cultures. PD0325901 or AZD6244 led to increased matrix protein expression (Col2a1 and Acan) and decreased expression of catabolic factors (Mmp13 and Adamts-5). Taken together, these data support the hypothesis that MEKi treatment can impact chondrocyte hypertrophy, matrix resorption, and fracture healing. These compounds can also affect bone architecture by expanding the hypertrophic zone of the growth plate and reducing osteoclast surface systemically.


Antithrombotic strategy in non-ST-segment elevation myocardial infarction patients undergoing percutaneous coronary intervention: insights from the ACTION (Acute Coronary Treatment and Intervention Outcomes Network) Registry.
The aim of this study was to examine the use of and outcomes associated with antithrombotic strategies in patients with non-ST-segment elevation myocardial infarction (NSTEMI) who undergo percutaneous coronary intervention (PCI). A variety of antithrombotic strategies have been tested in clinical trials for NSTEMI patients treated with PCI. Antithrombotic strategies for NSTEMI patients undergoing PCI at 217 ACTION (Acute Coronary Treatment and Intervention Outcomes Network) hospitals from January 1, 2007, to December 31, 2007, (n = 11,085) were classified into commonly observed antithrombotic groups: heparin alone (Hep alone; low-molecular-weight heparin or unfractionated heparin), bivalirudin alone (Bival alone), heparin with glycoprotein IIb/IIIa inhibitors (Hep/GPI), and bivalirudin with GPI (Bival/GPI). Baseline characteristics are shown across treatment groups. In addition, unadjusted and adjusted rates of in-hospital major bleeding and death are shown. The standard strategy used was Hep/GPI (64%), followed by Hep or Bival alone (28%), and Bival/GPI (8%). Patients who received Hep or Bival alone were older with more comorbidities, higher baseline bleeding and mortality risk, and lower peak troponin. Compared with patients who received Hep/GPI , those who received Hep alone and Bival alone had lower rates of major bleeding (adjusted odds ratio [OR]: 0.52; 95% confidence interval [CI]: 0.42 to 0.65; adjusted OR: 0.48; 95% CI: 0.39 to 0.60; respectively), yet only patients who received Bival alone had lower mortality (adjusted OR: 0.39; 95% CI: 0.21 to 0.71). NSTEMI patients undergoing PCI are more likely to receive Bival or Hep alone when at higher baseline bleeding risk than when at lower baseline bleeding risk. Despite higher baseline risk, those receiving Bival or Hep alone had less bleeding.


Prevalence of Chagas disease in Latin-American migrants living in Europe: a systematic review and meta-analysis.
Few studies have assessed the burden of Chagas disease in non-endemic countries and most of them are based on prevalence estimates from Latin American (LA) countries that likely differ from the prevalence in migrants living in Europe. The aim of this study was to systematically review the existing data informing current understanding of the prevalence of Chagas disease in LA migrants living in European countries. We conducted a systematic review and meta-analysis of studies reporting prevalence of Chagas disease in European countries belonging to the European Union (EU) before 2004 in accordance with the MOOSE guidelines and based on the database sources MEDLINE and Global Health. No restrictions were placed on study date, study design or language of publication. The pooled prevalence was estimated using random effect models based on DerSimonian & Laird method. We identified 18 studies conducted in five European countries. The random effect pooled prevalence was 4.2% (95%CI:2.2-6.7%); and the heterogeneity of Chagas disease prevalence among studies was high (I2 = 97%,p<0.001). Migrants from Bolivia had the highest prevalence of Chagas disease (18.1%, 95%CI:13.9-22.7%). Prevalence of Chagas in LA migrants living in Europe is high, particularly in migrants from Bolivia and Paraguay. Data are highly heterogeneous dependent upon country of origin and within studies of migrants from the same country of origin. Country-specific prevalence differs from the estimates available from LA countries. Our meta-analysis provides prevalence estimates of Chagas disease that should be used to estimate the burden of disease in European countries.


Power training and postmenopausal hormone therapy affect transcriptional control of specific co-regulated gene clusters in skeletal muscle.
At the moment, there is no clear molecular explanation for the steeper decline in muscle performance after menopause or the mechanisms of counteractive treatments. The goal of this genome-wide study was to identify the genes and gene clusters through which power training (PT) comprising jumping activities or estrogen containing hormone replacement therapy (HRT) may affect skeletal muscle properties after menopause. We used musculus vastus lateralis samples from early stage postmenopausal (50-57 years old) women participating in a yearlong randomized double-blind placebo-controlled trial with PT and HRT interventions. Using microarray platform with over 24,000 probes, we identified 665 differentially expressed genes. The hierarchical clustering method was used to assort the genes. Additionally, enrichment analysis of gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways was carried out to clarify whether assorted gene clusters are enriched with particular functional categories. The analysis revealed transcriptional regulation of 49 GO/KEGG categories. PT upregulated transcription in "response to contraction"-category revealing novel candidate genes for contraction-related regulation of muscle function while HRT upregulated gene expression related to functionality of mitochondria. Moreover, several functional categories tightly related to muscle energy metabolism, development, and function were affected regardless of the treatment. Our results emphasize that during the early stages of the postmenopause, muscle properties are under transcriptional modulation, which both PT and HRT partially counteract leading to preservation of muscle power and potentially reducing the risk for aging-related muscle weakness. More specifically, PT and HRT may function through improving energy metabolism, response to contraction as well as by preserving functionality of the mitochondria.


Topical use of dexpanthenol in skin disorders.
Pantothenic acid is essential to normal epithelial function. It is a component of coenzyme A, which serves as a cofactor for a variety of enzyme-catalyzed reactions that are important in the metabolism of carbohydrates, fatty acids, proteins, gluconeogenesis, sterols, steroid hormones, and porphyrins. The topical use of dexpanthenol, the stable alcoholic analog of pantothenic acid, is based on good skin penetration and high local concentrations of dexpanthenol when administered in an adequate vehicle, such as water-in-oil emulsions. Topical dexpanthenol acts like a moisturizer, improving stratum corneum hydration, reducing transepidermal water loss and maintaining skin softness and elasticity. Activation of fibroblast proliferation, which is of relevance in wound healing, has been observed both in vitro and in vivo with dexpanthenol. Accelerated re-epithelization in wound healing, monitored by means of the transepidermal water loss as an indicator of the intact epidermal barrier function, has also been seen. Dexpanthenol has been shown to have an anti-inflammatory effect on experimental ultraviolet-induced erythema. Beneficial effects of dexpanthenol have been observed in patients who have undergone skin transplantation or scar treatment, or therapy for burn injuries and different dermatoses. The stimulation of epithelization, granulation and mitigation of itching were the most prominent effects of formulations containing dexpanthenol. In double-blind placebo-controlled clinical trials, dexpanthenol was evaluated for its efficacy in improving wound healing. Epidermal wounds treated with dexpanthenol emulsion showed a reduction in erythema, and more elastic and solid tissue regeneration. Monitoring of transepidermal water loss showed a significant acceleration of epidermal regeneration as a result of dexpanthenol therapy, as compared with the vehicle. In an irritation model, pretreatment with dexpanthenol cream resulted in significantly less damage to the stratum corneum barrier, compared with no pretreatment. Adjuvant skin care with dexpanthenol considerably improved the symptoms of skin irritation, such as dryness of the skin, roughness, scaling, pruritus, erythema, erosion/fissures, over 3 to 4 weeks. Usually, the topical administration of dexpanthenol preparations is well tolerated, with minimal risk of skin irritancy or sensitization.


Humoral immune responses of white-tailed deer (Odocoileus virginianus) to Mycobacterium bovis BCG vaccination and experimental challenge with M. bovis.
Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple M. bovis antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (n=5), orally in liquid form (n=5), and subcutaneously (n=6) were evaluated over time after vaccination and after experimental challenge with virulent M. bovis and were compared to the responses by unvaccinated deer (n=6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple M. bovis antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials.


Brain MRI abnormalities in schizophrenia: same genes or same environment?
Structural brain volume abnormalities are among the most extensively studied endophenotypes in schizophrenia. Bivariate genetic model fitting (adjusted to account for selection) was used to quantify the genetic relationship between schizophrenia and brain volumes and to estimate the heritability of these volumes. We demonstrated by simulation that the adjusted genetic model produced unbiased estimates for endophenotype heritability and the genetic and environmental correlations. The model was applied to brain volumes (whole brain, hippocampus, third and lateral ventricles) in a sample of 14 monozygotic (MZ) twin pairs concordant for schizophrenia, 10 MZ discordant pairs, 17 MZ control pairs, 22 discordant sibling pairs, three concordant sibling pairs, and 114 healthy control subjects. Whole brain showed a substantial heritability (88%) and lateral ventricles substantial common environmental effects (67%). Whole brain showed a significant genetic correlation with schizophrenia, whereas lateral ventricles showed a significant individual specific correlation with schizophrenia. There were significant familial effects for hippocampus and third ventricle, but the analyses could not resolve whether these were genetic or environmental in origin (around 30%each). Using genetic model fitting on twin and sibling data we have demonstrated differential sources of covariation between schizophrenia and brain volumes, genetic in the case of whole brain volume and individual specific environment in the case of lateral ventricles.


Brown propolis attenuates cerebral ischemia-induced oxidative damage via affecting antioxidant enzyme system in mice.
Oxidative stress plays a critical role in ischemic brain injury. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) are the enzymes underlying the endogenous antioxidant mechanisms affected by stroke and are considered as oxidative stress biomarkers. Brown propolis (BP) is a bioactive natural product with a set of biological activities that in turn may differ depending on the area from which the substance is originated. The aim of this study was to investigate the effect of water-extracted brown propolis (WEBPs), from two regions of Iran, against cerebral ischemia-induced oxidative injury in a mouse model of stroke. Experimentally, the chemical characterization and total polyphenol content were determined using GC/MS and Folin-Ciocalteu assay respectively. Seventy-two adult male mice were randomly divided into the surgical sham group, control group (treated with vehicle), and four groups of WEBPs-treated animals. The WEBPs were administered at the doses of 100 and 200mg/kg IP, during four different time points. Oxidative stress biomarkers (SOD and GPx activity, SOD/GPx ratio), lipid peroxidation (LPO) index (malondialdehyde content) and infarct volume were measured 48h post stroke. Behavioral tests were evaluated 24 and 48h after stroke. WEBPs treatment resulted in significant restoration of antioxidant enzymes activity and a subsequent decrease in LPO as well as the infarct volume compared to the control group. Sensory-motor impairment and neurological deficits were improved significantly as well. These results indicate that Iranian BP confers neuroprotection on the stroke-induced neuronal damage via an antioxidant mechanism which seems to be mediated by the endogenous antioxidant system.


Developing senior hospital managers: does 'one size fit all'? - evidence from the evolving Chinese health system.
To improve the effectiveness and efficiency of health service provision in China, the National Health Commission has emphasised that training of all health service managers is essential. However, the implementation of that policy has proven challenging for various reasons, one of which is the lack of understanding of the competency requirements and gaps. The aims of the study were to develop an understanding of the characteristics and training experience of hospital managers in one major Chinese city, explore the difficulties they experience and relate them to their perceived importance of management competencies and the perceived level of their management competency. A cross-sectional, descriptive study with a three-component survey including the use of a validated management competency assessment tool was conducted with three senior executive groups (n = 498) from three categories of hospital in Jinan, Shandong Province, China. The survey confirmed that formal and informal management training amongst participants before commencing their management positions was inadequate. The core competencies identified in the Australia context were applicable to the management roles in Chinese hospitals. In addition, the senior executives had low levels of confidence in their management competence. Furthermore, the data showed significant differences between hospital categories and management levels in terms of their commitment to formal and informal training and self-perceived management competence. The study suggests that management training and support should be provided using a systematic approach with specific consideration to hospital types and management levels and positions. Such an approach should include clear competency requirements to guide management position recruitment and performance management.


Assessment of antiretroviral therapy knowledge and willingness of persons with HIV to support its uptake in Uganda.
Access to care and treatment services for human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) are hampered by human resource constraints and knowledge gaps about antiretroviral therapy. Training people with HIV/AIDS (PWA) as educators on antiretroviral therapy may help in the expansion of antiretroviral therapy-related knowledge in Africa. The aim of this study was to assess the antiretroviral therapy-specific knowledge, beliefs, and attitudes of PWA as well as their proactive communication with community members and to explore their willingness to serve as support personnel. Data were obtained from a large randomized trial of PWA. We analyzed qualitative and quantitative data from 524 PWA aged 18 years and over who initiated home-based antiretroviral therapy in 2003. We assessed knowledge and communication of HIV prevention and treatment messages by PWA to communities using structured messages complemented with other knowledge questions and the willingness of PWA to serve as support persons. Descriptive bivariate associations and logistic regression statistical methods were performed. In addition, qualitative data analysis was used. The level of knowledge about antiretroviral therapy was high among all PWA on several technical attributes. Overall, 90% of PWA reported that they had been consulted by community members for informed opinions on antiretroviral therapy, 70% felt they were opinion leaders on aspects of antiretroviral therapy within the communities, and approximately 70% were willing to be engaged as community support persons. Those who were classified as opinion leaders reported being approached more regularly by community members for expert advice about antiretroviral therapy compared with nonopinion leaders (odds ratio [OR] 11.7; 95% confidence interval [CI] 7.3-18.6), and opinion leaders were significantly more informed on most technical attributes of antiretroviral therapy, such as "who qualifies for antiretroviral therapy based on CD4 count" (OR 1.6, 95% CI 1.1-2.0) and "the need to be evaluated for antiretroviral therapy" (OR 1.8, 95% CI 1.2-2.0). Opinion leaders demonstrated correct knowledge and willingness to provide information on antiretroviral therapy care and treatment issues and were, in turn, consulted more frequently for antiretroviral therapy advice compared with nonopinion leaders. Training opinion leaders to work as community support personnel may increase knowledge about antiretroviral therapy in underserved communities.


A holistic investigation of the ecological correlates of abundance and body size for the endangered White's seahorse Hippocampus whitei.
Analysing the associations between the endangered White's seahorse Hippocampus whitei and characteristics of its environment (including habitat, prey and predator variables) in an estuary in New South Wales, Australia, revealed that seahorses had a greater number of significant associations with environmental correlates within a single seagrass bed than among seagrass beds. Predator abundance was negatively correlated with H. whitei abundances among seven seagrass beds (200-6,000 m apart) and no ecological correlate was associated with H. whitei body size distributions. Within the seagrass bed with the greatest number of H. whitei, individuals preferentially selected locations that were deeper, had denser seagrass, more epiphytic prey types and fewer predators. Smaller H. whitei were associated with greater depths within the bed. In this study, each class of ecological correlate (habitat, prey, predators) was found to have at least one significant relationship with H. whitei, depending on the scale, demonstrating that all three are important to H. whitei populations. As such, future studies that evaluate animal populations may benefit from holistic approaches that consider each of these together. For animals that are experiencing dramatic population declines due to habitat destruction, as H. whitei has over the last decade, a better understanding of its relationship to its environment is important to inform conservation action.


Assessing respiratory function depends on mechanical characteristics of balloon catheters.
Respiratory muscle function and lung and chest wall mechanics are reliably assessed by esophageal and gastric balloon catheters. The aim of this in vitro bench study was to assess the mechanical properties of commercially available balloon catheters using an experimental model with 3 defined compliances (27, 54, 90 mL/cm H2O). Six catheters were investigated in 4 conditions: (1) balloon pressure during initial inflation, (2) static pressure measurements at different filling volumes, (3) estimation of set compliances in the experimental lung model at different levels of superimposed pressure, and (4) elastic balloon properties after 16 h of inflation. 5/6 catheters showed initial pressure artifacts resulting from material adhesion. All static pressure measurements could be performed with an error < 1 cm H2O. Balloon overfilling resulted in larger errors in 4/6 catheters. Compliance determined from pressure measurements via the catheters differed by < 5% from that determined from direct pressure measurements. Sixteen hours of inflation resulted in a broader working range, that is, overfilling effects occurred at higher filling volumes. The reliability of pressure measurements and estimation of the lung model's compliance in the tested catheters are high. Filling volume appears to be critical for precise pressure measurement and compliance estimation. At first use, adhesion of the balloon material might prevent reliable pressure measurement.


Physiological and behavioural consequences of seizures induced in the rat by intrahippocampal tetanus toxin.
Injecting tetanus toxin into rat hippocampus induces a syndrome of intermittent generalized seizures which recurs for about one month. Following remission from their seizures, the rats exhibit very persistent impairments of learning and memory. Learning was impaired on a circular platform task and a spatial reference memory task, and evoked responses from the commissural-CA3 pyramidal cell system were depressed for up to 22 weeks after injection. There was no significant loss of pyramidal neurons because antidromic responses, evoked from other parts of the commissural fibre system, were not affected by the toxin treatment. The depression of these pyramidal neurons provides a reasonable physiological explanation for the learning impairment. These results suggest that impairments of neuronal function can be significant factors in the development of interictal behavioural abnormalities.


Chimeric proteins containing the cytoplasmic domains of the mannose 6-phosphate receptors codistribute with the endogenous receptors.
We have constructed and transiently expressed in HeLa cells a series of hybrid proteins in which the cytoplasmic domain or both the transmembrane and the cytoplasmic domains of the mannose 6-phosphate/insulin-like growth factor II receptor were fused to the ectodomain of the hemagglutinin of the influenza virus (HA), a typical plasma membrane protein. In addition, we have expressed a hybrid protein containing the luminal domain of HA fused to the transmembrane and cytoplasmic tail of the cation-dependent mannose 6-phosphate receptor. These hybrids were transported through and sorted from the secretory pathway as shown by acquisition of endo-H resistant oligosaccharides and their ability to recruit the Golgi assembly proteins AP-1 on the Golgi membrane. Like the mannose 6-phosphate receptors (MPRs), these hybrid proteins are also present in small amounts at the cell surface where they are likely to undergo endocytosis as disruption of the endocytosis signals contained in the MPR cytoplasmic domains induces their accumulation at the cell surface. Double immunofluorescence studies indicate that these chimeras codistribute with the endogenous MPRs at steady state. The results suggest that the cytoplasmic domains of the MPRs are sufficient to determine the steady-state distribution of the full-length proteins.


Developing an Inositol-Phosphate-Actuated Nanochannel System by Mimicking Biological Calcium Ion Channels.
In eukaryotic cells, ion channels, which ubiquitously present as polypeptides or proteins, usually regulate the ion transport across biological membranes by conformational switching of the channel proteins in response to the binding of diverse signaling molecules (e.g., inositol phosphate, abbreviated to InsP). To mimic the gating behaviors of natural Ca2+ channels manipulated by InsPs, a smart poly[(N-isopropylacrylamide-co-4-(3-acryloylthioureido) benzoic acid)0.2] (denoted as PNI-co-ATBA0.2) was integrated onto a porous anodic alumina (PAA) membrane, building an InsP-actuated nanochannel system. Driven by the intensive hydrogen bonding complexation of ATBA monomer with InsP, the copolymer chains displayed a remarkable and reversible conformational transition from a contracted state to a swollen one, accompanied with significant changes in surface morphology, wettability, and viscoelasticity. Benefiting from these features, dynamic gating behaviors of the nanochannels located on the copolymer-modified PAA membrane could be precisely manipulated by InsPs, reflected as a satisfactory linear relationship between real-time variation in transmembrane ionic current and the InsP concentration over a wide range from 1 nmol L-1 to 10 μmol L-1, as well as a clear discrimination among InsP2, InsP3, and InsP6. This study indicates the great potential of biomolecule-responsive polymers in the fabrication of biomimetic ion nanochannels and other nanoscale biodevices.


Laboratory diagnosis of factitious disorders.
Self-induced factitious disorders are defined and distinguished from conditions they may resemble. Review of the literature since 1965 indicates much more frequent reporting in recent years, but most health care providers are still not sufficiently aware of the common factitious disorder. Up to 5% of physician-patient encounters may be because of factitious disorders, but these are only suspected when the workup leads to contradictory findings. Laboratory tests are often the only definitive diagnostic method, and clinicians may not be familiar with current technologies. Some clinical clues are listed; heightened awareness and the need for early diagnosis are emphasized. Discordant laboratory results should raise the possibility of a factitious disorder. Sophisticated laboratory tests that can accurately assay very small amounts of specific hormones or foreign substances in body fluids facilitate the diagnosis. The primary physician can now confirm an initial clinical diagnosis of factitious disorder promptly and directly rather than only by exclusion.


Dielectrophoresis microsystem with integrated flow cytometers for on-line monitoring of sorting efficiency.
Dielectrophoresis (DEP) and flow cytometry are powerful technologies and widely applied in microfluidic systems for handling and measuring cells and particles. Here, we present a novel microchip with a DEP selective filter integrated with two microchip flow cytometers (FCs) for on-line monitoring of cell sorting processes. On the microchip, the DEP filter is integrated in a microfluidic channel network to sort yeast cells by positive DEP. The two FCs detection windows are set upstream and downstream of the DEP filter. When a cell passes through the detection windows, the light scattered by the cell is measured by integrated polymer optical elements (waveguide, lens, and fiber coupler). By comparing the cell counting rates measured by the two FCs, the collection efficiency of the DEP filter can be determined. The chips were used for quantitative determination of the effect of flow rate, applied voltage, conductivity of the sample, and frequency of the electric field on the sorting efficiency. A theoretical model for the capture efficiency was developed and a reasonable agreement with the experimental results observed. Viable and non-viable yeast cells showed different frequency dependencies and were sorted with high efficiency. At 2 MHz, more than 90% of the viable and less than 10% of the non-viable cells were captured on the DEP filter. The presented approach provides quantitative real-time data for sorting a large number of cells and will allow optimization of the conditions for, e.g., collecting cancer cells on a DEP filter while normal cells pass through the system. Furthermore, the microstructure is simple to fabricate and can easily be integrated with other microstructures for lab-on-a-chip applications.


The effects of stress and transcranial direct current stimulation (tDCS) on working memory: A randomized controlled trial.
Recent reviews of transcranial direct current stimulation (tDCS) show limited support for its initially cited enhancing effects on working memory (WM). They highlight the need for additional research, assessing the specific circumstances that optimize stimulation outcome. Social stress is an attractive candidate in this regard, as it affects WM and is mediated by prefrontal cortex activity; tDCS that targets these neuronal networks may, therefore, interact with social stress to affect WM. Our objective was to explore the interaction between social stress and tDCS on WM performance in a healthy cohort, 69 female participants were randomized to four experimental conditions (i.e., 2 × 2 design): stimulation (dlPFC tDCS vs. sham stimulation) and stress manipulation (Trier Social Stress Test [TSST] procedure vs. a friendly control TSST). Participants' attention, WM (assessed using an n-back task), and subjective/objective indicators of stress were assessed. A significant Stimulation × Stress Manipulation interaction was found, F(1, 65) = 6.208, p = .015, suggesting that active tDCS may increase WM performance in the no-stress conditions, while decreasing it under stress. Follow-up analyses of variance, however, were not significant (i.e., ps=.083 / .093), and Bayesian analyses were inconclusive. In conclusion, stress seems to be a crucial factor in determining the effects of tDCS, and tDCS may have an enhancing effect on WM at lower levels of stress, while being detrimental at higher stress levels (i.e., reversing the direction of effect). Possible theoretical underpinnings of the findings are discussed, while acknowledging the need for further research.


A Toxoplasma MORN1 null mutant undergoes repeated divisions but is defective in basal assembly, apicoplast division and cytokinesis.
The membrane occupation and recognition nexus protein 1 (MORN1) is highly conserved among apicomplexan parasites and is associated with several structures that have a role in cell division. Here we dissected the role of MORN1 using the relatively simple budding process of Toxoplasma gondii as a model. Ablation of MORN1 in a conditional null mutant resulted in pronounced defects suggesting a central role for MORN1 in apicoplast segregation and in daughter cell budding. Lack of MORN1 resulted in double-headed parasites. These Janus-headed parasites form two complete apical complexes but fail to assemble a basal complex. Moreover, these parasites were capable of undergoing several more budding rounds resulting in the formation of up to 16-headed parasites conjoined at the basal end. Despite this segregation defect, the mother's cytoskeleton was completely disassembled in every budding round. Overall this argues that successful completion of the budding is not required for cell cycle progression. None of the known basal complex components, including a set of recently identified inner membrane complex (IMC) proteins, localized correctly in these multi-headed parasites. These data suggest that MORN1 is essential for assembly of the basal complex, and that lack of the basal complex abolishes the contractile capacity assigned to the basal complex late in daughter formation. Consistent with this hypothesis we observe that MORN1 mutants fail to efficiently constrict and divide the apicoplast. We used the null background provided by the mutant to dissect the function of subdomains of the MORN1 protein. This demonstrated that deletion of a single MORN domain already prevented the function of MORN1 whereas a critical role for the short linker between MORN domains 6 and 7 was identified. In conclusion, MORN1 is required for basal complex assembly and loss of MORN1 results in defects in apicoplast division and daughter segregation.


Computer-assisted protein domain boundary prediction using the DomPred server.
Domain prediction from sequence is a particularly challenging task, and currently, a large variety of different methodologies are employed to tackle the task. Here we try to classify these diverse approaches into a number of broad categories. Completely automatic domain prediction from sequence alone is currently fraught with problems, but this should not be so surprising since human experts currently have significant disagreement on domain assignment even when given the structures. It can be argued that we should only test the domain prediction methods on benchmark data that human experts agree upon and this is the approach we take in this paper. Even for the data sets on which human experts agree, automatic structure-based domain assignment still cannot always agree, and so again it is still unlikely that domain prediction methods will reliably obtain correct results completely automatically. We make the argument that computer-assisted domain prediction is a more achievable goal. With this aim in mind, we present the DomPred server. This server provides the user with the results from two completely different categories of method (DPS and DomSSEA). In this paper, each method is individually benchmarked against one of the latest domain prediction benchmarks to provide information about their respective reliabilities. A variety of different benchmark scores are employed since the accuracy of a domain prediction method depends critically on what types of results one wishes to obtain (single/multi-domain classification, domain number, residue linker positions, etc.). Also both of these methods, implemented within the DomPred server, can suggest alternative domain predictions, allowing the user to make the final decision based on these results and applying their own background knowledge to the problem. The DomPred server is available from the URL:http://bioinf.cs.ucl.ac.uk/software.html.


Fitting mechanistic epidemic models to data: A comparison of simple Markov chain Monte Carlo approaches.
Simple mechanistic epidemic models are widely used for forecasting and parameter estimation of infectious diseases based on noisy case reporting data. Despite the widespread application of models to emerging infectious diseases, we know little about the comparative performance of standard computational-statistical frameworks in these contexts. Here we build a simple stochastic, discrete-time, discrete-state epidemic model with both process and observation error and use it to characterize the effectiveness of different flavours of Bayesian Markov chain Monte Carlo (MCMC) techniques. We use fits to simulated data, where parameters (and future behaviour) are known, to explore the limitations of different platforms and quantify parameter estimation accuracy, forecasting accuracy, and computational efficiency across combinations of modeling decisions (e.g. discrete vs. continuous latent states, levels of stochasticity) and computational platforms (JAGS, NIMBLE, Stan).


Full mutational mapping of titratable residues helps to identify proton-sensors involved in the control of channel gating in the Gloeobacter violaceus pentameric ligand-gated ion channel.
The Gloeobacter violaceus ligand-gated ion channel (GLIC) has been extensively studied by X-ray crystallography and other biophysical techniques. This provided key insights into the general gating mechanism of pentameric ligand-gated ion channel (pLGIC) signal transduction. However, the GLIC is activated by lowering the pH and the location of its putative proton activation site(s) still remain(s) unknown. To this end, every Asp, Glu, and His residue was mutated individually or in combination and investigated by electrophysiology. In addition to the mutational analysis, key mutations were structurally resolved to address whether particular residues contribute to proton sensing, or alternatively to GLIC-gating, independently of the side chain protonation. The data show that multiple residues located below the orthosteric site, notably E26, D32, E35, and D122 in the lower part of the extracellular domain (ECD), along with E222, H235, E243, and H277 in the transmembrane domain (TMD), alter GLIC activation. D122 and H235 were found to also alter GLIC expression. E35 is identified as a key proton-sensing residue, whereby neutralization of its side chain carboxylate stabilizes the active state. Thus, proton activation occurs allosterically to the orthosteric site, at the level of multiple loci with a key contribution of the coupling interface between the ECD and TMD.


Infection as a risk factor in the pathogenesis of primary biliary cirrhosis: pros and cons.
Primary biliary cirrhosis (PBC) is a chronic and slowly progressive cholestatic liver disease of autoimmune etiology, characterized by injury of the intrahepatic bile ducts that may eventually lead to cirrhosis and liver failure. Evidence suggests cardinal roles for both environmental factors and genetic susceptibility. Nevertheless, the absolute etiology of PBC is unclear, despite recent well-designed case-control studies that reported environmental risk factors, including infectious agents, for PBC. Of the reported infectious agents, some of them are not reproducible and remain controversial. However, infection is no doubt one of the major risks among the environmental factors. This is supported by the fact that infectious agents in autoimmune diseases express antigens resulting in molecular mimicry and xenobiotics that play a role in breaking tolerance. Taken together, recent findings from genome wide assays as well as novel animal models may enable us to better understand the mechanism of pathogenesis responsible for this disease.


The effects of barley-derived soluble fiber on serum lipids.
We wanted to determine the association between consumption of barley and changes in plasma lipids in healthy and hypercholesterolemic men and women. A systematic literature search was conducted from the earliest possible date through January 2008. Trials were included in the analysis if they were randomized controlled trials of barley that reported efficacy data on at least 1 lipid endpoint. A DerSimonian and Laird random-effects model was used in calculating the weighted mean difference (WMD) and its 95% confidence interval (CI). Statistical heterogeneity was addressed using the I(2) statistic. Visual inspection of funnel plots, Egger's weighted regression statistics, and the trim and fill method were used to assess for publication bias. We found 8 trials (n = 391 patients) of 4 to 12 weeks' duration evaluating the lipid-reducing effects of barley. The use of barley significantly lowered total cholesterol (weighted mean difference [WMD], -13.38 mg/dL; 95% CI, -18.46 to -8.31 mg/dL), low-density lipoprotein (LDL) cholesterol (WMD, -10.02 mg/dL; 95% CI, -14.03 to -6.00 mg/dL) and triglycerides (WMD, -11.83 mg/dL; 95% CI, -20.12 to -3.55 mg/dL) but did not appear to significantly alter high-density lipoprotein (HDL) cholesterol (P=.07). Barley-derived beta-glucan appears to beneficially affect total cholesterol, LDL-cholesterol, and triglycerides, but not HDL-cholesterol.


Chronic TNFα Exposure Induces Robust Proliferation of Olfactory Ensheathing Cells, but not Schwann Cells.
TNFα is persistently elevated in many injury and disease conditions. Previous reports of cytotoxicity of TNFα for oligodendrocytes and their progenitors suggest that the poor endogenous remyelination in patients with traumatic injury or multiple sclerosis may be due in part to persistent inflammation. Understanding the effects of inflammatory cytokines on potential cell therapy candidates is therefore important for evaluating the feasibility of their use. In this study, we assessed the effects of long term exposure to TNFα on viability, proliferation, migration and TNFα receptor expression of cultured rat olfactory ensheathing cells (OECs) and Schwann cells (SCs). Although OECs and SCs transplanted into the CNS produce similar myelinating phenotypes, and might be expected to have similar therapeutic uses, we report that they have very different sensitivities to TNFα. OECs exhibited positive proliferative responses to TNFα over a much broader range of concentrations than SCs. Low TNFα concentrations increased proliferation and migration of both OECs and SCs, but SC number declined in the presence of 100 ng/ml or higher concentrations of TNFα. In contrast, OECs exhibited enhanced proliferation even at high TNFα concentrations (up to 1 µg/ml) and showed no evidence of TNF cytotoxicity even at 4 weeks post-treatment. Furthermore, while both OECs and SCs expressed TNFαR1 and TNFαR2, TNFα receptor levels were downregulated in OECs after exposure to100 ng/ml TNFα for 5-7 days, but were either elevated or unchanged in SCs. These results imply that OECs may be a more suitable cell therapy candidate if transplanted into areas with persistent inflammation.