Phenotypic identification of over 1000 isolates of anaerobic bacteria recovered between 1999 and 2008 in a major Costa Rican hospital.
Because of limitations in infrastructure, the aetiology of infections caused by anaerobic bacteria is seldom determined in clinical laboratories of developing countries. This study reports on the identification of 1010 anaerobic bacterial isolates collected between 1999 and 2008 in a major Costa Rican hospital with the use of two commercial phenotypic systems (RapID 32A and API 20A). Approximately 60% of the isolates were Gram-positive and, among the 35 species of Gram-positive bacteria found, the genera Clostridium, Propionibacterium and Eggerthella, and anaerobic cocci predominated. Twenty eight species were found among 395 isolates of Gram-negative bacteria. Species of Bacteroides were very frequent, followed by species of Prevotella, Veillonella, Fusobacterium and Porphyromonas.


Prevalence and genetic diversity of Bartonella species isolated from wild rodents in Japan.
Here, we describe for the first time the prevalence and genetic properties of Bartonella organisms in wild rodents in Japan. We captured 685 wild rodents throughout Japan (in 12 prefectures) and successfully isolated Bartonella organisms from 176 of the 685 rodents (isolation rate, 25.7%). Those Bartonella isolates were all obtained from the rodents captured in suburban areas (rate, 51.8%), but no organism was isolated from the animals captured in city areas. Sequence analysis of rpoB and gltA revealed that the Bartonella isolates obtained were classified into eight genetic groups, comprising isolates closely related to B. grahamii (A-I group), B. tribocorum and B. elizabethae (B-J group), B. tribocorum and B. rattimassiliensis (C-K group), B. rattimassiliensis (D-L group), B. phoceensis (F-N group), B. taylorii (G-O group), and probably two additional novel Bartonella species groups (E-M and H-P). B. grahamii, which is one of the potential causative agents of human neuroretinitis, was found to be predominant in Japanese rodents. In terms of the relationships between these Bartonella genetic groups and their rodent species, (i) the A-I, E-M, and H-P groups appear to be associated with Apodemus speciosus and Apodemus argenteus; (ii) the C-K, D-L, and F-N groups are likely implicated in Rattus rattus; (iii) the B-J group seems to be involved in Apodemus mice and R. rattus; and (iv) the G-O group is probably associated with A. speciosus and Clethrionomys voles. Furthermore, dual infections with two different genetic groups of bartonellae were found in A. speciosus and R. rattus. These findings suggest that the rodent in Japan might serve as a reservoir of zoonotic Bartonella infection.


Relationship of splanchnic blood flow and portal venous resistance to elevated portal pressure in the dog.
To determine the relationship of splanchnic blood flow and portal venous resistance to elevated portal pressure, in situ perfusion of the splanchnic circuit was carried out in 12 freshly killed dogs at varying perfusion rates and degrees of acute and chronic portal vein constriction. In six normal dogs before being killed, portal pressure averaged 8-8 +/- 1-2 mmHg and portal flow 658 +/- 81 ml/min. In the absence of portal vein constriction, increasing splanchnic perfusion to 1800 ml/min, minimally raised portal pressure (12-8 +/- 1-5 mmHg). With progressive acute constriction of the portal vein, however, comparable perfusion rates lead to progressively higher portal pressure levels and with greater than 90% constriction, portal pressure was greater than 30 mmHg with minimal elevation in splanchnic flow rate. In six other dogs before being killed but live to nine weeks after placement of an ameroid constrictor on the portal vein, portal pressure averaged 13-6 +/- 1-4 mmHg or slightly higher than in normal dogs (P greater than 0-02). Mesenteric venography and necropsy findings uniformly demonstrated 90% occlusion of the portal vein with extensive portasystemic collateralization. With increased perfusion of the splanchnic bed, portal pressure rose rapidly to approximately 35 mmHg with a flow rate of 1 800 ml/min.. These data suggest that, in disorders where resistance to transhepatic portal flow is marked, a small increment in splanchnic blood flow, which normally exerts little or no influence on portal pressure, promotes profound portal hypertension and may account for spontaneous 'rupture' of oesophageal varices. On the other hand, where resistance to transhepatic portal flow is mild but splanchnic blood flow is markedly hyperdynamic, reducing inflow may be sufficient to ameliorate extreme portal hypertension and its sequelae.


Texture descriptors and voxels for the early diagnosis of Alzheimer's disease.
Early and accurate diagnosis of Alzheimer's Disease (AD) is critical since early treatment effectively slows the progression of the disease thereby adding productive years to those afflicted by this disease. A major problem encountered in the classification of MRI for the automatic diagnosis of AD is the so-called curse-of-dimensionality, which is a consequence of the high dimensionality of MRI feature vectors and the low number of training patterns available in most MRI datasets relevant to AD. A method for performing early diagnosis of AD is proposed that combines a set of SVMs trained on different texture descriptors (which reduce dimensionality) extracted from slices of Magnetic Resonance Image (MRI) with a set of SVMs trained on markers built from the voxels of MRIs. The dimension of the voxel-based features is reduced by using different feature selection algorithms, each of which trains a separate SVM. These two sets of SVMs are then combined by weighted-sum rule for a final decision. Experimental results show that 2D texture descriptors improve the performance of state-of-the-art voxel-based methods. The evaluation of our system on the four ADNI datasets demonstrates the efficacy of the proposed ensemble and demonstrates a contribution to the accurate prediction of AD. Ensembles of texture descriptors combine partially uncorrelated information with respect to standard approaches based on voxels, feature selection, and classification by SVM. In other words, the fusion of a system based on voxels and an ensemble of texture descriptors enhances the performance of voxel-based approaches.


Examination of cardiovascular risk factors and rurality in Appalachian children.
The prevalence of childhood cardiovascular disease (CVD) risk factors often increases in more rural geographic regions in the USA. However, research on the topic often has conflicting results. Researchers note differences in definitions of rurality and other factors that would lead to differences in inference, including appropriate use of statistical clustering analysis, representative data, and inclusion of individual-level covariates. The present study's objective was to examine CVD risk factors during childhood by geographic distribution in the US Appalachian region as a first step towards understanding the health disparities in this area. Rurality and CVD risk factors (including blood pressure, body-mass index (BMI), and cholesterol) were examined in a large, representative sample of fifth-grade students (N=73 014) from an Appalachian state in the USA. A six-category Rural-Urban Continuum Codes classification system was used to define rurality regions. Mixed modeling analysis was used to appropriately cluster individuals within 725 unique zip codes in each of these six regions, and allowed for including several individual-level socioeconomic factors as covariates. Rural areas had better outcomes for certain CVD risk factors (lowest low-density lipoprotein cholesterol (LDL-C), and blood pressure (BP) and highest high-density lipoprotein cholesterol (HDL-C)) whereas mid-sized metro and town areas presented with the worst CVD risk factors (highest BMI% above ideal, mean diastolic BP, LDL-C, total cholesterol, triglyceride levels and lowest HDL-C) outcomes in children and adolescence in this Appalachian state. Counter to the study hypothesis, mid-sized metro areas presented with the worst CVD risk factors outcomes in children and adolescence in the Appalachian state. This data contradicts previous literature suggesting a straightforward link between rurality and cardiovascular risk factors. Future research should include a longitudinal design and explore some of the mechanisms between cardiovascular risk factors and rurality.


Failure of daily tenofovir to prevent HIV transmission or the establishment of a significant viral reservoir despite continued antiretroviral therapy.
Truvada is licenced for HIV-1 prevention in the USA and is available in the private sector. Tenofovir performed as well as Truvada in the PARTNERS PrEP study and is used as HIV pre-exposure prophylaxis (PreP) in some settings. The clinical efficacy of Tenofovir for PrEP outside a clinical trial is unknown. Antiretroviral therapy (ART) at acute HIV-1 infection (AHI) limits the size of the reservoir, optimizing the chance of maintaining viral control off therapy. As such ART at acute HIV infection is proposed to offer a functional cure in a minority of subjects. We present two cases where Tenofovir PrEP failed to prevent HIV acquisition and failed to limit viral reservoir. Two individuals receiving tenofovir monotherapy for Hepatitis B monoinfection were diagnosed with AHI as defined by a negative HIV antibody test within three months of a positive HIV test following unsafe sex with casual male partners. In-depth histories were taken. Viral genotypes and Tenofovir drug levels were measured from samples taken as close to HIV seroconversion as possible and subsequent samples were analyzed for proviral Total HIV-1 DNA by qPCR. Patient A had received tenofovir for the preceding six years and always maintained an undetectable Hepatitis B viral load with no concerns about adherence. Two weeks preceding the positive HIV antibody test, he experienced mild symptoms (fever, pharyngitis) of HIV seroconversion. HIV status was confirmed by a repeat fourth generation HIV antibody test and by Western Blot and an HIV viral load was undetectable. Tenofovir trough level at HIV diagnosis was within normal limits. The regimen was intensified to Eviplera and a total HIV-1 DNA was 1381 copies/million CD4 T cells. Patient B received four regimens for hepatitis B treatment before starting tenofovir monotherapy in 2011 and subsequently maintained an undetectable hepatitis B viral load. After three years of tenofovir monotherapy he developed a severe symptomatic seroconversion illness and tested HIV antibody positive. The baseline HIV viral load was 103,306 copies/mL. The regimen was intensified and total HIV-1 DNA was 2746 copies/million CD4 T cells. Further investigation into the efficacy of tenofovir for PrEP outside a clinical trial is required. ART at AHI does not always lead to a low viral reservoir. To explore the possibility of replication incompetent virus, viral outgrowth assays are underway.


Improving clustering with metabolic pathway data.
It is a common practice in bioinformatics to validate each group returned by a clustering algorithm through manual analysis, according to a-priori biological knowledge. This procedure helps finding functionally related patterns to propose hypotheses for their behavior and the biological processes involved. Therefore, this knowledge is used only as a second step, after data are just clustered according to their expression patterns. Thus, it could be very useful to be able to improve the clustering of biological data by incorporating prior knowledge into the cluster formation itself, in order to enhance the biological value of the clusters. A novel training algorithm for clustering is presented, which evaluates the biological internal connections of the data points while the clusters are being formed. Within this training algorithm, the calculation of distances among data points and neurons centroids includes a new term based on information from well-known metabolic pathways. The standard self-organizing map (SOM) training versus the biologically-inspired SOM (bSOM) training were tested with two real data sets of transcripts and metabolites from Solanum lycopersicum and Arabidopsis thaliana species. Classical data mining validation measures were used to evaluate the clustering solutions obtained by both algorithms. Moreover, a new measure that takes into account the biological connectivity of the clusters was applied. The results of bSOM show important improvements in the convergence and performance for the proposed clustering method in comparison to standard SOM training, in particular, from the application point of view. Analyses of the clusters obtained with bSOM indicate that including biological information during training can certainly increase the biological value of the clusters found with the proposed method. It is worth to highlight that this fact has effectively improved the results, which can simplify their further analysis.The algorithm is available as a web-demo at http://fich.unl.edu.ar/sinc/web-demo/bsom-lite/. The source code and the data sets supporting the results of this article are available at http://sourceforge.net/projects/sourcesinc/files/bsom.


Characterization of the terminal stages of chlorophyll (ide) synthesis in etioplast membrane preparations.
1. Chlorophyll (ide) formation from protochlorophyll (ide) that is normally inactive was demonstrated in etioplast membranes isolated from maize and barlley plants, the process being dependent on intermittent illumination and the addition of NADPH. 2. The addition of NADPH to the membranes was shown to result in the conversion of inactive protochlorophyll (ide) absorbing at about 630 nm into a form(s) with light-absorption maxima at about 640 and 652 nm, both of which disappear when chlorophyll (ide) is formed on illumination. 3. The temperature-dependence of the activation process and its response to a variety of reagents were examined. From these, the conclusion is drawn that -SH groups are involved in the activation but in the active complex these are unavailable for reaction with -SH reagents. 4. Evidence is presented for the occurrence of glucose 6-phosphate dehydrogenase activity within etioplasts and the suggestion is made that the oxidative pentose phosphate pathway can provide the NADPH required for chlorophyll biosynthesis during the early stages of greening.


11beta-hydroxysteroid dehydrogenase type 1 and obesity.
The metabolic syndrome consists of a constellation of co-associated metabolic abnormalities such as insulin resistance, type 2 diabetes, dyslipidaemia, hypertension and visceral obesity. For many years endocrinologists have noted the striking resemblance between this disease state and that associated with Cushing's syndrome. However, in the metabolic syndrome plasma cortisol levels tend to be normal or lower than in normal individuals. Nevertheless there is strong evidence that glucocorticoid action underlies metabolic disease, largely from rodent obesity models where removing glucocorticoids reverses obesity and its metabolic abnormalities. The apparent paradox of similar metabolic defects - despite the opposing plasma glucocorticoid profiles of Cushing's and idiopathic metabolic syndrome - remained intriguing until the discovery that intracellular glucocorticoid reactivation was elevated in adipose tissue of obese rodents and humans. The enzyme that mediates this activation, conversion of cortisone (11-dehydrocorticosterone in rodents) to cortisol (corticosterone in rodents), locally within tissues is 11beta -hydroxysteroid dehydrogenase type 1 (11beta -HSD1). In order to determine whether elevated tissue 11beta -HSD1 contributed to obesity and metabolic disease, transgenic mice overexpressing 11beta -HSD1 in adipose tissue or liver were made. Adipose-selective 11beta -HSD1 transgenic mice exhibited elevated intra-adipose and portal, but not systemic corticosterone levels, abdominal obesity, hyperglycaemia, insulin resistance, dyslipidaemia and hypertension. In contrast, transgenic overexpression of 11beta -HSD1 in liver yielded an attenuated metabolic syndrome with mild insulin resistance, dyslipidaemia, hypertension and fatty liver, but not obesity or glucose intolerance. Together with early data using non-selective 11beta -HSD1 inhibitors to insulin sensitise humans, this corroborated the notion that the enzyme may be a good therapeutic target in the treatment of the metabolic syndrome. Further, a transgenic model of therapeutic 11beta -HSD1 inhibition, 11beta -HSD1 gene knock-out (11beta -HSD1-/-) mice, exhibited improved glucose tolerance, a 'cardioprotective' lipid profile, reduced weight gain and visceral fat accumulation with chronic high-fat feeding. Recent evidence further suggests that high fat-mediated downregulation of adipose 11beta -HSD1 may be an endogenous pathway that underpins adaptive disease resistance in genetically predisposed mouse strains. This mechanism could feasibly make up a genetic component of innate obesity resistance in humans. The efficacy of 11beta -HSD1 inhibitors has recently been extended to include increased energy expenditure and reduction of arteriosclerosis, and therefore may be of significant therapeutic value in the metabolic syndrome, with complementary effects upon liver adipose tissue, muscle, pancreas and plaque-prone vessels.


Initial clinical results of stereotactic radiotherapy for the treatment of craniopharyngiomas.
The efficacy and toxicity of stereotactic radiotherapy (SRT) for the treatment of craniopharyngioma has been retrospectively evaluated in 16 patients. The median tumor diameter was 2.8 cm (range 1.5-6.1) and the median tumor volume was 7.7 cc (range 0.7-62.8). SRT was delivered to a single isocenter using a dedicated 6 MV linear accelerator to patients immobilized with a relocatable stereotactic head frame. The three-year actuarial overall survival was 93% and the rate of survival free of any imaging evidence of progressive disease was 75%. The three-year actuarial survival rates free of solid tumor growth or cyst enlargement were 94% and 81% respectively. Our results suggest that SRT is a safe and effective treatment approach for patients with craniopharyngioma. Long-term follow-up is required to determine whether the normal tissue-sparing inherent with SRT results in reduction of the neurocognitive effects of conventional radiotherapy for craniopharyngioma. SRT can be delivered to craniopharyngioma that may be difficult to treat with stereotactic radiosurgery due to proximity of the optic chiasm. Further clinical experience is necessary to determine the clinical utility of beam shaping in the setting of SRT.


When caterpillars attack: biogeography and life history evolution of the Miletinae (Lepidoptera: Lycaenidae).
Of the four most diverse insect orders, Lepidoptera contains remarkably few predatory and parasitic species. Although species with these habits have evolved multiple times in moths and butterflies, they have rarely been associated with diversification. The wholly aphytophagous subfamily Miletinae (Lycaenidae) is an exception, consisting of nearly 190 species distributed primarily throughout the Old World tropics and subtropics. Most miletines eat Hemiptera, although some consume ant brood or are fed by ant trophallaxis. A well-resolved phylogeny inferred using 4915 bp from seven markers sampled from representatives of all genera and nearly one-third the described species was used to examine the biogeography and evolution of biotic associations in this group. Biogeographic analyses indicate that Miletinae likely diverged from an African ancestor near the start of the Eocene, and four lineages dispersed between Africa and Asia. Phylogenetic constraint in prey selection is apparent at two levels: related miletine species are more likely to feed on related Hemiptera, and related miletines are more likely to associate with related ants, either directly by eating the ants, or indirectly by eating hemipteran prey that are attended by those ants. These results suggest that adaptations for host ant location by ovipositing female miletines may have been retained from phytophagous ancestors that associated with ants mutualistically.


A favorable clinical trend is associated with CD8 T-cell immune responses to the human papillomavirus type 16 e6 antigens in women being studied for abnormal pap smear results.
The goal of this study was to examine the role of CD8 T-cell responses to human papillomavirus type 16 (HPV-16) in a favorable clinical trend in women being studied for abnormal Pap smear results. Human papillomavirus-deoxyribonucleic acid testing and enzyme-linked immunospot assay using the HPV-16 E6 and E7 antigens were performed. The subjects with subsequent normal histologic diagnoses were considered to be "regressors" (n = 28), whereas those with histologic diagnoses of cervical intraepithelial neoplasia 1, 2, or 3 were considered to have short-term persistence of cervical abnormality and were designated to be "persistors" (n = 37). There was a higher percentage of CD8 T-cell responses to the E6 antigen in the regressors (15/28 or 53.6%) when compared with the persistors (10/37 or 27.0%; p = .04), but there was no recorded response difference for the E7 antigen. Results were the same when the analyses for E6 included only subjects who were high-risk HPV-positive (p = .01). The CD8 T-cell immune responses to the HPV-16 E6 antigens but not to E7 antigens are associated with a favorable clinical trend regardless of HPV types currently detected.


Consequences of extensive habitat fragmentation in landscape-level patterns of genetic diversity and structure in the Mediterranean esparto grasshopper.
Anthropogenic habitat fragmentation has altered the distribution and population sizes in many organisms worldwide. For this reason, understanding the demographic and genetic consequences of this process is necessary to predict the fate of populations and establish management practices aimed to ensure their viability. In this study, we analyse whether the spatial configuration of remnant semi-natural habitat patches within a chronically fragmented landscape has shaped the patterns of genetic diversity and structure in the habitat-specialist esparto grasshopper (Ramburiella hispanica). In particular, we predict that agricultural lands constitute barriers to gene flow and hypothesize that fragmentation has restricted interpopulation dispersal and reduced local levels of genetic diversity. Our results confirmed the expectation that isolation and habitat fragmentation have reduced the genetic diversity of local populations. Landscape genetic analyses based on circuit theory showed that agricultural land offers ∽1000 times more resistance to gene flow than semi-natural habitats, indicating that patterns of dispersal are constrained by the spatial configuration of remnant patches of suitable habitat. Overall, this study shows that semi-natural habitat patches act as corridors for interpopulation gene flow and should be preserved due to the disproportionately large ecological function that they provide considering their insignificant area within these human-modified landscapes.


A pilot study of deep sclerectomy with implantation of chromic suture material as a collagen implant: medium-term results.
A pilot study of the safety and efficacy of deep sclerectomy with a chromic suture implant (DSCI) for open angle glaucoma. This prospective trial involved 23 eyes of 16 consecutive patients with uncontrolled open angle glaucoma. The DSCI procedure involved radial suturing of a 2 mm piece of 1/0 chromic suture as an implant. Intraocular pressure (IOP) measurements and eye examinations were performed preoperatively and regularly postoperatively for up to 66 months. The mean follow-up period was 36.3 (SD 19.9) months. The mean preoperative IOP was 26.3 (SD 8.5) mmHg. The mean postoperative IOP was 14.1 (SD 2.8) mmHg at month 12, and 16.5 (SD 2.3) mmHg at 36 months. Complete success rate, defined as an IOP lower than 21 mmHg without medication was 77% of eyes at 36 months. Qualified success rate, defined as an IOP lower than 21 mmHg with medication, was 100% at 36 months. Paired t-test and Wilcoxon test analysis of preoperative vs postoperative IOP at 36 months indicated a significant reduction in IOP. Intraoperative complications included rupture of the trabecular meshwork in seven eyes. Exclusion of these patients from the results analysis revealed a mean follow-up of 37.1 months (SD 9), preoperative mean IOP was 25.9 mmHg (SD 4.1), 12-month mean IOP was 13.4 mmHg (SD 2.8), and 36-month mean IOP 16.6 mmHg (SD 2.1). Early postoperative complications included hyphaema in two eyes and dislocation of implant in one eye. One eye developed a central retinal vein occlusion and cataract progression occurred in one eye. The mean postoperative duration of use of topical dexamethasone was 5.4 months (SD 2). DSCI using a chromic suture implant provides good control of IOP at medium-term follow-up with few postoperative complications. Prolonged use of postoperative anti-inflammatory medication was necessary.


Air pollution and short-term clinical outcomes of patients with acute myocardial infarction.
Ambient air pollution is well-known to be a serious risk factor for cardiovascular diseases, stroke, and death. However, the association between air pollutants (AP) exposure and short-term clinical outcomes in acute myocardial infarction (AMI) patients (pts) has not been elucidated well. In the present study, 37 880 AMI pts were enrolled from October 2005 to December 2013 in a nationwide large-scale, prospective, multicentre Korea AMI registry (KAMIR registry; http://www.kamir.or.kr). We obtained data on AP (e.g., NO2 , SO2 , CO, O3 and PM10 ) from the Korean National Institute of Environmental Research (NIER; http://www.nier.go.kr). Clinical endpoints included death, recurrent myocardial infarction (Re-MI), any revascularization and composite of all-cause death and Re-MI. Exposure to AP is defined as the average exposure to AP within 24 hours before AMI admission. We observed that a 0.01 part per million (ppm) increase in NO2 concentration, 0.001 ppm increase in SO2 concentration, and 0.1 ppm increase in CO concentration each increased the risk of total death by 9.7% (95% CI, 6.2%-13.4%), 1.9% (95% CI, 0.3%-3.6%), and 2.1% (95% CI, 0.5%-3.9%), respectively. Exceptionally, O3 decreased the risk of total death by 0.6% (95% CI -0.2% to -1.0%) per 0.01 ppm increase. PM10 was not related to any cardiovascular events. AP were each stratified into five quintiles according to ranges of AP levels. After adjusting analysis for risk variables, only high quintiles (Q4, Q5) of NO2 were positively associated with total death, cardiac death and MI, while SO2 , CO, O3 and PM10 were shown to be not related to any cardiovascular events at all levels. In AMI patients, each AP and its concentration has shown a different effect to short-term mortality and cardiovascular events.


The relation between calcium intake and body composition in a Dutch population: The Amsterdam Growth and Health Longitudinal Study.
To investigate whether dietary calcium intake is related to body mass index and the sum of four skinfolds among subjects in the Amsterdam Growth and Health Longitudinal Study (the Netherlands), the authors followed a cohort of men and women from age 13 years in 1977 to age 36 years in 2000. Longitudinal linear regression analyses were performed with generalized estimating equations in continuous and categorical models, with adjustment for possible confounders. Results showed that calcium intake during adolescence is a weak predictor of calcium intake in adulthood. In this population, only a slight indication was found of a weak inverse relation of calcium intake with body composition. No differences were observed between the middle (800-1,200 mg/day) and high (>1,200 mg/day) groups of calcium intake, suggesting a threshold of approximately 800 mg/day above which calcium intake has no additional beneficial effect on body composition.


Longitudinal variations of brain functional connectivity: A case report study based on a mouse model of epilepsy.
Brain Functional Connectivity (FC) quantifies statistical dependencies between areas of the brain. FC has been widely used to address altered function of brain circuits in control conditions compared to different pathological states, including epilepsy, a major neurological disorder. However, FC also has the as yet unexplored potential to help us understand the pathological transformation of the brain circuitry. Our hypothesis is that FC can differentiate global brain interactions across a time-scale of days. To this end, we present a case report study based on a mouse model for epilepsy and analyze longitudinal intracranial electroencephalography data of epilepsy to calculate FC changes from the initial insult (status epilepticus) and over the latent period, when epileptogenic networks emerge, and at chronic epilepsy, when unprovoked seizures occur as spontaneous events. We found that the overall network FC at low frequency bands decreased immediately after status epilepticus was provoked, and increased monotonously later on during the latent period. Overall, our results demonstrate the capacity of FC to address longitudinal variations of brain connectivity across the establishment of pathological states.


Improving translatability of preclinical studies for neuromuscular disorders: lessons from the TREAT-NMD Advisory Committee for Therapeutics (TACT).
Clinical trials for rare neuromuscular diseases imply, among other investments, a high emotional burden for the whole disease community. Translation of data from preclinical studies to justify any clinical trial must be carefully pondered in order to minimize the risk of clinical trial withdrawal or failure. A rigorous distinction between proof-of-concept and preclinical efficacy studies using animal models is key to support the rationale of a clinical trial involving patients. This Review evaluates the experience accumulated by the TREAT-NMD Advisory Committee for Therapeutics, which provides detailed constructive feedback on clinical proposals for neuromuscular diseases submitted by researchers in both academia and industry, and emphasizes that a timely critical review of preclinical efficacy data from animal models, including biomarkers for specific diseases, combined with adherence to existing guidelines and standard protocols, can significantly help to de-risk clinical programs and prevent disappointments and costly engagement.


Clinical outcomes of noninfectious sternal dehiscence after median sternotomy.
Infectious complications of median sternotomy carry significant morbidity and mortality. However, the outcomes of noninfectious sternal dehiscence have not been addressed. We have identified the preoperative characteristics, postoperative complications, and long-term functional outcomes of patients after reoperation for noninfectious sternal dehiscence and compared these patients with a control group to determine risk factors for dehiscence. Retrospective review of the cardiac surgery database identified 48 patients with noninfectious sternal dehiscence in a group of 12,380 median sternotomies between 1994 and 2004. The review included diagnosis, demographics, concomitant medical conditions, and surgical outcomes. Functional outcomes were assessed using the Short Form-12 questionnaire. One hundred fifty-six median sternotomy patients served as controls. Follow-up was 97.9% (47 of 48 patients) complete, for a total of 150.1 patient-years. Mean age of patients at reoperation was 58.8 +/- 12.8 years, with a male to female ratio of 45:3. Multivariate analysis determined that New York Heart Association class IV, obesity, and chronic obstructive pulmonary disease were preoperative risk factors for sternal dehiscence. The incidence of sternal dehiscence was 0.39% at a mean interval between initial operation and reoperation of 5.4 months. At a mean interval of 3.9 months, 14.6% (7 of 48) of patients required additional sternal procedures. Infectious complications after reoperation occurred in 12.5% (6 of 48). Functional outcomes demonstrated that 72.2% (26 of 36) had no or mild limitation of physical activities, with 90.5% (38 of 42) reporting no or mild sternal pain at follow-up. Although patients undergoing surgical correction of noninfectious sternal dehiscence fare better than those with infectious complications, optimal sternal approximation during the initial procedure and sternal precautions during convalescence should be emphasized to prevent recurrent complications.


Thymidine phosphorylase expression in normal and hyperplastic endometrium.
To investigate the expression of thymidine phosphorylase (TP), a known angiogenic factor for endothelial cells, in normally cycling endometrium and various forms of endometrial hyperplasia. TP expression was assessed with the P-GF.44C monoclonal antibody, using the alkaline phosphatase anti-alkaline phosphatase method. Ninety two normal and hyperplastic endometria were studied. In normal proliferative endometrium, TP is found exclusively in the basal layer and the inner third of the functionalis; expression is cytoplasmic in glandular epithelium and nuclear in stromal cells. It is invariably patchy. This immunohistochemical picture remains almost unaltered during the early and mid secretory phase of the normal menstrual cycle but, most impressively, TP is expressed uniformly in the epithelium of all endometrial glands towards the end of the cycle. At this stage, expression is mixed nuclear/cytoplasmic and there is very little stromal nuclear staining. In simple endometrial hyperplasia, the staining pattern for TP is identical to normal proliferative endometrium, with a distribution that is usually limited to a few rather weakly proliferating glands and to the adjacent periglandular stroma of the deep endometrium. The distribution is more extensive in complex and atypical endometrial hyperplasias, where a mixed nuclear/cytoplasmic pattern usually prevails over the pure cytoplasmic reaction. TP is expressed consistently in normal and hyperplastic endometrium, suggesting a role in physiological and pathological angiogenesis. In normal endometrium, TP has a definite pattern of distribution, which is dependent on the phase of the menstrual cycle, whereas in all forms of endometrial hyperplasia the enzyme is randomly distributed and lacks an orderly pattern.


Human laboratory paradigms in alcohol research.
Human laboratory studies have a long and rich history in the field of alcoholism. Human laboratory studies have allowed for advances in alcohol research in a variety of ways, including elucidating neurobehavioral mechanisms of risk, identifying phenotypically distinct subtypes of alcohol users, investigating the candidate genes underlying experimental phenotypes for alcoholism, and testing mechanisms of action of alcoholism pharmacotherapies on clinically relevant translational phenotypes, such as persons exhibiting positive-like alcohol effects or alcohol craving. Importantly, the field of human laboratory studies in addiction has progressed rapidly over the past decade and has built upon earlier findings of alcohol's neuropharmacological effects to advancing translational research on alcoholism etiology and treatment. To that end, the new generation of human laboratory studies has focused on applying new methodologies, further refining alcoholism phenotypes, and translating these findings to studies of alcoholism genetics, medication development, and pharmacogenetics. The combination of experimental laboratory approaches with the recent developments in neuroscience and pharmacology has been particularly fruitful in furthering our understanding of the impact of individual differences in alcoholism risk and in treatment response. This review of the literature focuses on human laboratory studies of subjective intoxication, alcohol craving, anxiety, and behavioral economics. Each section discusses opportunities for phenotype refinement under laboratory conditions, as well as its application to translational science of alcoholism. A summary and recommendations for future research are also provided.


Activation of the hexosamine pathway by glucosamine in vivo induces insulin resistance of early postreceptor insulin signaling events in skeletal muscle.
To explore potential cellular mechanisms by which activation of the hexosamine pathway induces insulin resistance, we have evaluated insulin signaling in conscious fasted rats infused for 2-6 h with saline, insulin (18 mU x kg(-1) x min(-1)), or insulin and glucosamine (30 micromol x kg(-1) x min(-1)) under euglycemic conditions. Glucosamine infusion increased muscle UDP-N-acetylglucosamine concentrations 3.9- and 4.3-fold over saline- or insulin-infused animals, respectively (P < 0.001). Glucosamine induced significant insulin resistance to glucose uptake both at the level of the whole body and in rectus abdominis muscle, and it blunted the insulin-induced increase in muscle glycogen content. At a cellular level, these metabolic effects were paralleled by inhibition of postreceptor insulin signaling critical for glucose transport and glycogen storage, including a 45% reduction in insulin-stimulated insulin receptor substrate (IRS)-1 tyrosine phosphorylation (P = 0.02), a 44% decrease in IRS-1 association with the p85 regulatory subunit of phosphatidylinositol (PI) 3-kinase (P = 0.03), a 34% reduction in IRS-1-associated PI 3-kinase activity (P = 0.03), and a 51% reduction in insulin-stimulated glycogen synthase activity (P = 0.03). These alterations in postreceptor insulin signaling were time-dependent and paralleled closely the progressive inhibition of systemic glucose disposal from 2 to 6 h of glucosamine infusion. We also demonstrated that glucosamine infusion results in O-linked N-acetylglucosamine modification of IRS-1 and IRS-2. These data indicate that activation of the hexosamine pathway may directly modulate early postreceptor insulin signal transduction, perhaps via posttranslation modification of IRS proteins, and thus contribute to the insulin resistance induced by chronic hyperglycemia.


Cell line name recognition in support of the identification of synthetic lethality in cancer from text.
The recognition and normalization of cell line names in text is an important task in biomedical text mining research, facilitating for instance the identification of synthetically lethal genes from the literature. While several tools have previously been developed to address cell line recognition, it is unclear whether available systems can perform sufficiently well in realistic and broad-coverage applications such as extracting synthetically lethal genes from the cancer literature. In this study, we revisit the cell line name recognition task, evaluating both available systems and newly introduced methods on various resources to obtain a reliable tagger not tied to any specific subdomain. In support of this task, we introduce two text collections manually annotated for cell line names: the broad-coverage corpus Gellus and CLL, a focused target domain corpus. We find that the best performance is achieved using NERsuite, a machine learning system based on Conditional Random Fields, trained on the Gellus corpus and supported with a dictionary of cell line names. The system achieves an F-score of 88.46% on the test set of Gellus and 85.98% on the independently annotated CLL corpus. It was further applied at large scale to 24 302 102 unannotated articles, resulting in the identification of 5 181 342 cell line mentions, normalized to 11 755 unique cell line database identifiers. The manually annotated datasets, the cell line dictionary, derived corpora, NERsuite models and the results of the large-scale run on unannotated texts are available under open licenses at http://turkunlp.github.io/Cell-line-recognition/. email@example.com.


Monoubiquitination of Syntaxin 5 Regulates Golgi Membrane Dynamics during the Cell Cycle.
The Golgi apparatus undergoes a ubiquitin-dependent disassembly and reassembly process during each cycle of cell division. Here we report the identification of the Golgi t-SNARE syntaxin 5 (Syn5) as the ubiquitinated substrate. Syn5 is monoubiquitinated by the ubiquitin ligase HACE1 in early mitosis and deubiquitinated by the deubiquitinase VCIP135 in late mitosis. Syn5 ubiquitination on lysine 270 (K270) in the SNARE domain impairs the interaction between Syn5 and the cognate v-SNARE Bet1 but increases its binding to p47, the adaptor protein of p97. Expression of the Syn5 K270R mutant in cells impairs post-mitotic Golgi reassembly. Therefore, monoubiquitination of Syn5 in early mitosis disrupts SNARE complex formation. Subsequently, ubiquitinated Syn5 recruits p97/p47 to the mitotic Golgi fragments and promotes post-mitotic Golgi reassembly upon ubiquitin removal by VCIP135. Overall, this study reveals both the substrate and the mechanism of ubiquitin-mediated regulation of Golgi membrane dynamics during the cell cycle.


A2A adenosine receptor protects tumors from antitumor T cells.
The A2A adenosine receptor (A2AR) has been shown to be a critical and nonredundant negative regulator of immune cells in protecting normal tissues from inflammatory damage. We hypothesized that A2AR also protects cancerous tissues by inhibiting incoming antitumor T lymphocytes. Here we confirm this hypothesis by showing that genetic deletion of A2AR in the host resulted in rejection of established immunogenic tumors in approximately 60% of A2AR-deficient mice with no rejection observed in control WT mice. The use of antagonists, including caffeine, or targeting the A2 receptors by siRNA pretreatment of T cells improved the inhibition of tumor growth, destruction of metastases, and prevention of neovascularization by antitumor T cells. The data suggest that effects of A2AR are T cell autonomous. The inhibition of antitumor T cells via their A2AR in the adenosine-rich tumor microenvironment may explain the paradoxical coexistence of tumors and antitumor immune cells in some cancer patients (the "Hellstrom paradox"). We propose to target the hypoxia-->adenosine-->A2AR pathway as a cancer immunotherapy strategy to prevent the inhibition of antitumor T cells in the tumor microenvironment. The same strategy may prevent the premature termination of immune response and improve the vaccine-induced development of antitumor and antiviral T cells. The observations of autoimmunity during melanoma rejection in A2AR-deficient mice suggest that A2AR in T cells is also important in preventing autoimmunity. Thus, although using the hypoxia-->adenosine-->A2AR pathway inhibitors may improve antitumor immunity, the recruitment of this pathway by selective drugs is expected to attenuate the autoimmune tissue damage.


Scarf versus chevron osteotomy for the correction of 1-2 intermetatarsal angle in hallux valgus: a systematic review and meta-analysis.
The chevron and scarf osteotomies are commonly used for the surgical management of hallux valgus (HV). However, there is debate as to whether one osteotomy provides more 1-2 intermetatarsal (1-2 IMA) correction than the other. The objective of this systematic review and meta-analysis was to compare the effectiveness of 3 types of first metatarsal osteotomy for reducing the 1-2 IMA in HV correction: the chevron osteotomy, the long plantar arm (modified) chevron osteotomy, and the scarf osteotomy. A systematic search for eligible studies was performed of the following databases: Medline, Embase (Ovid), CINAHL (EBSCO Host), and The Cochrane Database of Systematic Reviews and Cochrane Central Register of Controlled Clinical Trials. Only English-language studies previous to May 2010 were included in the review. Additional hand and electronic content searches of relevant foot and orthopaedic journals were performed. Criteria for inclusion in this analysis included systematic reviews of randomized controlled trials, prospective and retrospective cohort studies, and case-control studies, as well as case-series studies involving the chevron, scarf, or long plantar arm chevron osteotomy of >20 participants with a minimum of 80% follow-up. Quality of evidence of the included studies was assessed with the Grading of Recommendations Assessment, Development and Evaluation system. All pooled analyses were based on a fixed effects model. There was a total of 1351 participants who underwent either a chevron (n = 1028), scarf (n = 300), or long plantar arm chevron osteotomy (n = 23). Only one study for the long plantar arm chevron group fitted the eligibility criteria for this review; however, it was not amenable to meta-analysis. The chevron osteotomy was associated with a mean reduction of 1-2 IMA from preoperative to postoperative of 5.33° (95% confidence interval, 5.12 to 5.54, p < .001), and the scarf osteotomy was associated with a mean reduction of 6.21° (95% confidence interval, 5.70 to 6.72, p < .001). There was a statistically significant 0.88° increase in the correction of the 1-2 IMA in favor of the scarf osteotomy compared with the chevron osteotomy. The studies included in this review were of very low- to low-quality evidence. Our findings indicate that the scarf osteotomy provides greater correction of the 1-2 IMA when used for HV correction. However, only a weak recommendation in favor of the scarf osteotomy can be made based on the low quality of evidence of the studies included in this analysis.


MDR1 Ala893 polymorphism is associated with inflammatory bowel disease.
Crohn disease (CD) and ulcerative colitis (UC) are overlapping chronic inflammatory bowel diseases (IBDs). Suggestive evidence for linkage at chromosome 7q has been reported for both CD and UC. Contained within this region is the gene for MDR1 (multidrug resistance), a membrane transport protein for which human polymorphisms have been reported in Ala893Ser/Thr and C3435T that alter pharmacokinetic profiles for a variety of drugs. Because mdr1 knockout mice spontaneously develop colitis, exonic regions were resequenced and tested for IBD association in a large, multicenter North American cohort. Two missense mutations, Asn21Asp and Ala893Ser/Thr, as well as the expression-associated polymorphism C3435T, described elsewhere, were genotyped in the entire cohort. Significant association of Ala893 with IBD was observed by both case-control analysis (P=.002) and the pedigree disequilibrium test (PDT [P=.00020-.00030]) but not for the Asn21Asp or C3435T polymorphisms. Significant association by PDT was observed within the subset with CD (P=.0014-.00090), with similar, nonsignificant trends in a smaller subset with UC. The Ala893Ser/Thr variant is triallelic, and the associated, common allele is Ala893, with undertransmission of the 893Ser (common) and the 893Thr (rare) variants. The Ala893 variant has decreased activity compared with the 893Ser variant; therefore, the association with human IBD is consistent with the murine model of mdr1 deficiency. Taken together, these data support the association of the common Ala893 polymorphism with IBD specifically and, more broadly, provides additional support for its contribution to interindividual pharmacogenetic variation.


[Genital tuberculosis: a rare cause of postmenopausal bleeding].
Tuberculosis remains a global health problem, being the genitourinary tract the second most common site for tuberculosis infection after the lungs. Genital tuberculosis is now undergoing a worrying recrudescence. We report two cases of postmenopausal women who presented with vaginal bleeding. General physical and gynecological examination detected no abnormality. Transvaginal pelvic ultrasound showed fluid in the endometrial cavity in both cases. The office hysteroscopy was suggestive of focal endometrial thickness and an endometrial biopsy was performed. The histopathologic examination of biopsies found epithelioid cell granulomas without malignant cells. Culture of the endometrium was positive for Mycobacterium tuberculosis. Involvement of other systems was not detected. The patients started receiving antituberculosis treatment. Genital tuberculosis is rare in postmenopausal women and responsible for only approximately 1% of postmenopausal bleeding. However it is a curable disease and an early diagnosis is important and may prevent unnecessary invasive procedures for the patient.


Maximising Lucerne (Medicago sativa) Pasture Intake of Dairy Cows: 2-the Effect of Post-Grazing Pasture Height and Mixed Ration Level.
The effects of lucerne (Medicago sativa) post-grazing residual pasture height on pasture utilisation (vertical and horizontal), pasture intake and animal production were investigated in a sub-tropical partial mixed ration dairy system. The study took place at the Gatton Research Dairy, Southeast Queensland (-27.552, 152.333), with a 26-day adaptation period followed by two 8-day measurement periods during August and September 2018. A quantity of 30 multiparous Holstein-Friesian dairy cows were offered two levels of mixed ration, 7 and 14 kg dry matter (DM)/cow/day for low and high levels respectively, and five levels of pasture allocation, to achieve decreasing residual pasture heights. Pasture allocations measured from 5 cm above ground level for the low mixed ration groups averaged 12.7, 15.9, 19.8, 35.3 and 49.2 kg DM/cow/day, and for the high mixed ration groups averaged 5.0, 8.3, 10.3, 18.6, and 25.2 kg DM/cow/day, respectively. As pasture allocation decreased, cows were forced to graze further down into the pasture sward, and therefore residual pasture height declined. Total intake (kg DM/cow/day) declined as residual pasture height (expressed as % of the initial height) declined, irrespective of mixed ration level, decreasing by 0.5 kg DM/cow/day for every 10% decrease in residual pasture height. Low total intakes were associated with high non-esterified fatty acid (NEFA) levels in plasma, indicating mobilisation of fat tissue to maintain milk production. In the high allocation treatments, an area of pasture remained ungrazed and cows were only grazing the top leafy stratum where pasture intake rate and intake were highest. Therefore, to maximise intake in sub-tropical partial mixed ration (PMR) systems, lucerne pasture should be allocated so that cows are always grazing the top leafy stratum. This can be achieved by ensuring the pasture around faecal patches remains ungrazed.


An interferon lambda 4-associated variant in the hepatitis C virus RNA polymerase affects viral replication in infected cells.
Host IFNL4 haplotype status contributes to the development of chronic hepatitis C virus (HCV) infection in individuals who are acutely infected with the virus. In silico studies revealed that specific amino acid variants at multiple sites on the HCV polyprotein correlate with functional single-nucleotide polymorphisms (SNPs) in the IFNL4 locus. Thus, SNPs at the IFNL4 locus may select variants that influence virus replication and thereby the outcome of infection. Here, we examine the most significantly IFNL4-associated amino acid variants that lie in the 'lambda (L) 2 loop' of the HCV NS5B RNA polymerase. L2 loop variants were introduced into both sub-genomic replicon and full-length infectious clones of HCV and viral replication was examined in the presence and absence of exogenous IFNλ4. Our data demonstrate that while mutation of the NS5B L2 loop affects replication, individual IFNL4-associated variants have modest but consistent effects on replication in both the presence and absence of IFNλ4. Given the strong genetic association between these variants and IFNL4, these data suggest a nuanced effect of each individual position on viral replication, the combined effect of which might mediate resistance to the effects of IFNλ4.


Evolution of Evidence and Guideline Recommendations for the Medical Management of Severe Traumatic Brain Injury.
Brain Trauma Foundation (BTF) Guidelines for medical management of severe traumatic brain injury (TBI) have become a global standard for the treatment of TBI patients. We aim to explore the evolution of the guidelines for the management of severe TBI. We reviewed the four editions of the BTF guidelines published over the past 20 years. The 1996 and 2000 editions were merged because of minimal differences, and are referred to as the 1996 edition. We described changes in topics and recommendations over time, and analyzed predictors of survival of recommendations with logistical regression. The guidelines contained 27 recommendations on 18 topics in 2016, 35 recommendations on 15 topics in 2007, and 22 recommendations on 10 topics in 1996. Substantial delays were found between the search for evidence and the guideline publication, ranging from 18 to 34 months. The overall body of evidence comprised 189 studies on 18 topics in 2016, compared with 156 studies on 15 topics in 2007 and 180 studies on 10 topics in 1996. Over time, a total of 175 studies were discarded from the evidence base following more rigorous grading of evidence. A total of 15/23 (65%) of the 1996/2000 recommendations were discarded over time. Out of 12 new recommendations introduced in the 2007 edition, 8 (66%) were discarded in 2016. Survival of recommendations varied between 33% and 100% for level I recommendations and between 11% and 31% for level II and III recommendations. No predictors of survival of recommendations were found. Substantial delays exist between literature search and publication, and survival rate of TBI guideline recommendations is poor. These factors may adversely affect currency and adherence to guidelines. The TBI community should take responsibility for improving the quality of the evidence base and ensuring that the translation of the evidence into guidelines supports clinicians in daily clinical practice.


Consensus guideline for the diagnosis and treatment of aromatic l-amino acid decarboxylase (AADC) deficiency.
Aromatic L-amino acid decarboxylase deficiency (AADCD) is a rare, autosomal recessive neurometabolic disorder that leads to a severe combined deficiency of serotonin, dopamine, norepinephrine and epinephrine. Onset is early in life, and key clinical symptoms are hypotonia, movement disorders (oculogyric crisis, dystonia, and hypokinesia), developmental delay, and autonomic symptoms.In this consensus guideline, representatives of the International Working Group on Neurotransmitter Related Disorders (iNTD) and patient representatives evaluated all available evidence for diagnosis and treatment of AADCD and made recommendations using SIGN and GRADE methodology. In the face of limited definitive evidence, we constructed practical recommendations on clinical diagnosis, laboratory diagnosis, imaging and electroencephalograpy, medical treatments and non-medical treatments. Furthermore, we identified topics for further research. We believe this guideline will improve the care for AADCD patients around the world whilst promoting general awareness of this rare disease.


Psychiatric hospitalisation and suicide among the very old in Denmark: population-based register study.
Very old people have higher suicide rates than the younger elderly population. Psychiatric disorders are known to have a strong association with suicide among elderly people. To analyse the analyse the suicide risk associated with psychiatric hospitalisation among the very old (> or =80 years) compared with the middle-aged (50-64 years) and old (65-79 years) populations. Individual-level data on the entire Danish population aged 50 years or over were analysed for the period 1994-1998. Relative suicide risks were calculated using event-history analysis. Among 1978 527 persons, 2323 died by suicide. Although the very old group exhibited a four-fold to five-fold increase in risk of suicide for those previously hospitalised, we noted an inverse interaction effect: the increase is distinctly smaller compared with that in the middle-aged and old groups. The association between suicide and psychiatric hospitalisation is much weaker for the very old than for the old. Psychiatric disorders among very old people may be interacting with other disorders, may be underdiagnosed or treated in other healthcare settings.


Distinct genomic aberrations associated with ERG rearranged prostate cancer.
Emerging molecular and clinical data suggest that ETS fusion prostate cancer represents a distinct molecular subclass, driven most commonly by a hormonally regulated promoter and characterized by an aggressive natural history. The study of the genomic landscape of prostate cancer in the light of ETS fusion events is required to understand the foundation of this molecularly and clinically distinct subtype. We performed genome-wide profiling of 49 primary prostate cancers and identified 20 recurrent chromosomal copy number aberrations, mainly occurring as genomic losses. Co-occurring events included losses at 19q13.32 and 1p22.1. We discovered three genomic events associated with ERG rearranged prostate cancer, affecting 6q, 7q, and 16q. 6q loss in nonrearranged prostate cancer is accompanied by gene expression deregulation in an independent dataset and by protein deregulation of MYO6. To analyze copy number alterations within the ETS genes, we performed a comprehensive analysis of all 27 ETS genes and of the 3 Mbp genomic area between ERG and TMPRSS2 (21q) with an unprecedented resolution (30 bp). We demonstrate that high-resolution tiling arrays can be used to pin-point breakpoints leading to fusion events. This study provides further support to define a distinct molecular subtype of prostate cancer based on the presence of ETS gene rearrangements.


Comparative studies of the effects of quinuronium sulfate and diminazene diaceturate in sheep.
Biochemical parameters in 20 sheep were investigated following administration of quinuronium sulfate or diminazene diaceturate. The usual signs of salivation, micturition, anorexia, depression, muscular tremors and ataxia were observed within 20 min in sheep receiving therapeutic and double doses of quinuronium sulfate. There was an increased dose dependent plasma LDH activity above baseline values following administration of quinuronium sulfate and a non-dose dependent increased trend in diminazene diaceturate treatment. Plasma CK activity had an increased trend (p less than 0.05) above baseline values following administration of the two babesicides. Plasma BUN levels increased significantly (p less than 0.05) following administration of the two drugs. This study indicated that quinuronium sulfate is more organotoxic and hypotensive than diminazene diaceturate at therapeutic and/or above therapeutic dosages.


Antegrade common bile duct (CBD) stenting after laparoscopic CBD exploration.
Laparoscopic common bile duct exploration (LCBDE) has been found to be a safe, efficient and cost-effective treatment for choledocholithiasis. Following LCBDE, the clearance may be ascertained by a cholangiogram or choledochoscopy. The common bile duct (CBD) may be closed primarily with or without a stent in situ or may be drained by means of a T-tube or a biliary enteric anastomosis. In our series of 464 patients of choledocholithiasis, 100 patients underwent closure of the CBD with an indwelling antegrade stent following LCBDE. LCBDE was performed by direct massage of CBD, saline lavage, direct pickup with choledocholithotomy forceps or by basketing. Fragmentation of impacted stones in situ was performed in a few patients. Completion choledochoscopy was performed by means of a pediatric bronchoscope. A 10-cm, 7 Fr. double-flap biliary stent was placed in situ after LCBDE. There was no mortality in the series. There was no conversion either. The median duration of the operation was 75 min. The mean postoperative hospital stay was 3.5 days. One patient had a minor postoperative biliary leak. One patient had a right sub-hepatic collection. Four patients developed postoperative port infection. The stents were removed endoscopically after 4 weeks. Sixty-eight patients could be followed up till 1 year. There has been no incidence of residual disease and the patients on follow-up are asymptomatic. In our experience, a single stage laparoscopic treatment of cholelithiasis with choledocholithiasis is a safe, viable and cost-effective option. Closure of the CBD over an antegrade stent is a feasible option but requires advanced skills in minimal access surgical techniques, especially endosuturing. The procedure may be performed safely in expert hands without mortality and with negligible morbidity.


Tripin/hSgo2 recruits MCAK to the inner centromere to correct defective kinetochore attachments.
hSgo2 (previously annotated as Tripin) was recently reported to be a new inner centromere protein that is essential for centromere cohesion (Kitajima et al., 2006). In this study, we show that hSgo2 exhibits a dynamic distribution pattern, and that its localization depends on the BUB1 and Aurora B kinases. hSgo2 is concentrated at the inner centromere of unattached kinetochores, but extends toward the kinetochores that are under tension. This localization pattern is reminiscent of MCAK, which is a microtubule depolymerase that is believed to be a key component of the error correction mechanism at kinetochores. Indeed, we found that hSgo2 is essential for MCAK to localize to the centromere. Delocalization of MCAK accounts for why cells depleted of hSgo2 exhibit kinetochore attachment defects that go uncorrected, despite a transient delay in the onset of anaphase. Consequently, these cells exhibit a high frequency of lagging chromosomes when they enter anaphase. We confirmed that hSgo2 is associated with PP2A, and we propose that it contributes to the spatial regulation of MCAK activity within inner centromere and kinetochore.


Genome-wide mapping of alternative splicing in Arabidopsis thaliana.
Alternative splicing can enhance transcriptome plasticity and proteome diversity. In plants, alternative splicing can be manifested at different developmental stages, and is frequently associated with specific tissue types or environmental conditions such as abiotic stress. We mapped the Arabidopsis transcriptome at single-base resolution using the Illumina platform for ultrahigh-throughput RNA sequencing (RNA-seq). Deep transcriptome sequencing confirmed a majority of annotated introns and identified thousands of novel alternatively spliced mRNA isoforms. Our analysis suggests that at least approximately 42% of intron-containing genes in Arabidopsis are alternatively spliced; this is significantly higher than previous estimates based on cDNA/expressed sequence tag sequencing. Random validation confirmed that novel splice isoforms empirically predicted by RNA-seq can be detected in vivo. Novel introns detected by RNA-seq were substantially enriched in nonconsensus terminal dinucleotide splice signals. Alternative isoforms with premature termination codons (PTCs) comprised the majority of alternatively spliced transcripts. Using an example of an essential circadian clock gene, we show that intron retention can generate relatively abundant PTC(+) isoforms and that this specific event is highly conserved among diverse plant species. Alternatively spliced PTC(+) isoforms can be potentially targeted for degradation by the nonsense mediated mRNA decay (NMD) surveillance machinery or regulate the level of functional transcripts by the mechanism of regulated unproductive splicing and translation (RUST). We demonstrate that the relative ratios of the PTC(+) and reference isoforms for several key regulatory genes can be considerably shifted under abiotic stress treatments. Taken together, our results suggest that like in animals, NMD and RUST may be widespread in plants and may play important roles in regulating gene expression.


Diagnostic Sensitivity of Different Applications of Pathergy Test for Behçet's Disease.
This study aims to compare the sensitivity of the intradermal application of the pathergy test with the "three-step pathergy test". The study included 60 patients with Behçet's disease (BD) (23 males, 37 females; mean age 33.9±9.7 years; range, 11 to 57 years) and 100 patients with recurrent aphthous stomatitis as the control group (39 males, 61 females; mean age 32.4±12.1 years; range, 11 to 66 years). Simultaneous intradermal pathergy test and three-step pathergy test were applied to each patient. Twenty-six patients (43.3%) with BD had positive three-step pathergy test and 18 (30%) of them had positive intradermal pathergy test. A statistically significant relationship was found between the three-step pathergy test positivity and male sex while no relationship was detected between the disease activity and the positivity of the pathergy test. In the three-step pathergy test, pathergy positivity with intramuscular method was statistically significantly higher than with intravenous method. The three-step pathergy test in patients with BD was found to have higher sensitivity compared with the intradermal pathergy test. The intramuscularly applied pathergy test was the main cause of increase in sensitivity. Further studies are needed to develop new applications of pathergy test that would increase the sensitivity and which are also easy to apply.


Novel predictors of sepsis outperform the American Burn Association sepsis criteria in the burn intensive care unit patient.
The purpose of this study was to determine whether systemic inflammatory response syndrome (SIRS) and American Burn Association (ABA) criteria predict sepsis in the burn patient and develop a model representing the best combination of novel clinical sepsis predictors. A retrospective, case-controlled, within-patient comparison of burn patients admitted to a single intensive care unit from January 2005 to September 2010 was made. Blood culture results were paired with documented sepsis: positive-sick, negative-sick (collectively defined as sick), and negative-not sick. Data for all predictors were collected for the 72 hours before blood culture. Variables were evaluated using regression and area under the curve (AUC) analyses. Fifty-nine subjects represented 177 culture periods. SIRS criteria were not discriminative: 98% of the subjects met criteria. ABA sepsis criteria were different on the day before (P = .004). The six best-fit variables identified for the model included heart rate > 130 beats per min, mean arterial pressure < 60 mm Hg, base deficit < -6 mEq/L, temperature < 36°C, use of vasoactive medications, and glucose > 150 mg/dl. The model was significant in predicting positive-sick and sick, with an AUC of 0.775 (P < .001) and 0.714 (P < .001), respectively; comparatively, the ABA criteria AUC was 0.619 (P = .028) and 0.597 (P = .035), respectively. Usefulness of the ABA criteria to predict sepsis is limited to the day before blood culture is obtained. A significant contribution of this research is the identification of six novel sepsis predictors for the burn patient.


Repeat HIV testing practices in the era of HIV self-testing among adults in KwaZulu-Natal, South Africa.
Repeat HIV testing is important in high HIV burden communities to enable sustainability of prevention initiatives; however, an understanding of repeat testing practices is limited. Additional HIV testing approaches may be required to increase testing. HIV self-testing is an additional testing approach, but knowledge on its potential for repeat testing is limited. This study explored repeat HIV testing practices and uptake of HIV self-testing among repeat testers, following exposure to HIV self-testing. HIV testing practices were explored at two time points. During Phase 1, eighty in-depth interviews were conducted among 40 consenting adults, and 30 telephonic contacts were completed during Phase 2. Framework analysis was used to analyse the transcripts from the in-depth interviews. The practice of repeat HIV testing is primarily influenced by HIV status awareness and risk exposure. Thirteen regular testers and one HIV naïve tester at baseline had undergone repeat testing through the use of a traditional testing approach such as HIV counselling and testing as reported in Phase 2. HIV self-testing has a role among repeat testers, but affordability and access are barriers.


Spontaneous remission of acromegaly after infarctive apoplexy with a possible relation to MRI and diabetes mellitus.
Pituitary apoplexy is a rare clinical syndrome associated with rapid enlargement of a pituitary mass. We report the initial presentation, subsequent course and outcome of an acromegalic patient who developed spontaneous remission following pituitary apoplexy with pathologic findings of tumor infarction. A 38 year-old man with typical acromegalic features was referred to our hospital. He had been diabetic and hypertensive. His basal GH and IGF-1 levels were high (80 µg/L and 747 ng/mL respectively). Sella MRI showed a macroadenoma about 19×20 mm in size. He admitted to emergency department with complains of severe frontal headache accompanied by nausea and vomiting two days after MRI was taken. His neurological examination and visual field test were normal. Emergent MRI of the sella disclosed an enhancing intrasellar mass of 24×23 mm compressing the optic chiasm. The patient underwent transsphenoidal decompression of the lesion. Histological examination revealed an adenomatous tissue showing nonhemorrhagic coagulation necrosis. Before surgery, his GH levels declined to 2.72 µg/L spontaneously and after surgery he was in remission even leading to a state of growth hormone deficiency. When apoplexy occurs in functioning adenomas, it may cause spontaneous remission. However pituitary apoplexy due to tumor infarction is very rare. Various precipitating factors have been reported in 25-30% of pituitary apoplexy patients. Diabetes mellitus and diabetic ketoacidosis are one of these. The presence of contrast media induced endothelial swelling with the result of hypoperfusion and diabetes mellitus associated vasculopathy might be a precipitating factor in this patient.


Whole-animal metabolic rate is a repeatable trait: a meta-analysis.
Repeatability studies are gaining considerable interest among physiological ecologists, particularly in traits affected by high environmental/residual variance, such as whole-animal metabolic rate (MR). The original definition of repeatability, known as the intraclass correlation coefficient, is computed from the components of variance obtained in a one-way ANOVA on several individuals from which two or more measurements are performed. An alternative estimation of repeatability, popular among physiological ecologists, is the Pearson product-moment correlation between two consecutive measurements. However, despite the more than 30 studies reporting repeatability of MR, so far there is not a definite synthesis indicating: (1) whether repeatability changes in different types of animals; (2) whether some kinds of metabolism are more repeatable than others; and most important, (3) whether metabolic rate is significantly repeatable. We performed a meta-analysis to address these questions, as well as to explore the historical trend in repeatability studies. Our results show that metabolic rate is significantly repeatable and its effect size is not statistically affected by any of the mentioned factors (i.e. repeatability of MR does not change in different species, type of metabolism, time between measurements, and number of individuals). The cumulative meta-analysis revealed that repeatability studies in MR have already reached an asymptotical effect size with no further change either in its magnitude and/or variance (i.e. additional studies will not contribute significantly to the estimator). There was no evidence of strong publication bias.


The effect of confinement on the crystalline microstructure of polymer : fullerene bulk heterojunctions.
We investigate the effect of confinement on the crystalline microstructure of the polymer component of polymer : fullerene bulk heterojunction thin films using grazing incidence wide angle X-ray scattering. We find that the polymer crystallite size decreases and the alignment of the molecules along the surface normal increases, as the thin-film thickness is reduced from 920 nm to <20 nm and approaches the thin-film confinement regime. Furthermore, we find that the polymer crystallite size near the surface (air interface) is lower than the crystallite size in the bulk or the bottom (substrate interface) of bulk heterojunction films thicker than the confinement regime. Variation in polymer crystallite size can cause changes in charge carrier mobility and recombination rates, which in turn affect the performance of bulk heterojunction thin film devices such as photovoltaics and photodetectors.


[Evaluation of the posture of patients before and after orthodontic surgery].
The term "craniomandibular disorder" is used to describe a series of symptoms and signs that directly affect the stomatognathic apparatus with possible repercussions on the otovestibular and oculomotor apparatus and on the cervical spine that may condition the entire body posture. The aim of this study was to evaluate a series of parameters correlated to the occlusal situation and to verify how these factors are affected by a change following the correction of occulusal ratios. The paper reports a series of 15 patients suffering from facial dysmorphia treated surgically in the Division of Maxillofacial Surgery of S. Giovanni Battista Hospital in Turin in which an evaluation was made of posture and symptoms linked to craniomandibular disorders before surgery and six months after. At the preoperative evaluation a high incidence of this type of symptoms and signs was observed, and in particular a high percentage of patients with postural imbalance and asymmetry of the frontal body segments. From the results obtained it emerges that the surgical re-adaptation of occlusion is followed by esthetic improvement and also by a good recovery of frontal postural symmetry in conformity with the reduced frequency and intensity of symptoms classified as craniomandibular disorders.


Reduced Expression Level of Protein Phosphatase PPM1E Serves to Maintain Insulin Secretion in Type 2 Diabetes.
Reversible phosphorylation is an important regulatory mechanism. Regulation of protein phosphorylation in β-cells has been extensively investigated, but less is known about protein dephosphorylation. To understand the role of protein dephosphorylation in β-cells and type 2 diabetes (T2D), we first examined mRNA expression of the type 2C family (PP2C) of protein phosphatases in islets from T2D donors. Phosphatase expression overall was changed in T2D, and that of PPM1E was the most markedly downregulated. PPM1E expression correlated inversely with HbA1c. Silencing of PPM1E increased glucose-stimulated insulin secretion (GSIS) in INS-1 832/13 cells and/or islets from patients with T2D, whereas PPM1E overexpression decreased GSIS. Increased GSIS after PPM1E silencing was associated with decreased oxidative stress, elevated cytosolic Ca2+ levels and ATP to ADP ratio, increased hyperpolarization of the inner mitochondrial membrane, and phosphorylation of CaMKII, AMPK, and acetyl-CoA carboxylase. Silencing of PPM1E, however, did not change insulin content. Increased GSIS, cell viability, and activation of AMPK upon metformin treatment in β-cells were observed upon PPM1E silencing. Thus, protein dephosphorylation via PPM1E abrogates GSIS. Consequently, reduced PPM1E expression in T2D may be a compensatory response of β-cells to uphold insulin secretion under metabolic duress. Targeting PPM1E in β-cells may thus represent a novel therapeutic strategy for treatment of T2D.


[Two multidrug-resistant Salmonella infantis isolates behave like hypo-invasive strains but have high intracellular proliferation].
In this work, plasmid-encoded virulence factors in two Salmonella Infantis isolates carrying multiresistance plasmids were investigated. In addition, their invasion and proliferative ability in non-phagocytic cells was studied. None of them showed the typical determinants of virulence plasmids (spy operon). The invasion assays of S. Infantis isolates on eukaryotic cells showed a decreased ability to invade but they remained and proliferated in the cytoplasm regardless of having used a permissive (HeLa) or non-permissive (NRK) cell line. Finally, there was no microscopic evidence suggesting a bactericidal effect of these eukaryotic cell lines on the isolates tested.


Fast flip-flop of halogenated cobalt bis(dicarbollide) anion in a lipid bilayer membrane.
Transmembrane translocation (flip-flop) of cobalt bis(dicarbollide) (COSAN) anions, elicited by application of a voltage-jump across the lipid bilayer membrane, manifested itself in monoexponential electrical current transients in the microsecond time scale. Halogenation of COSAN led to multi-fold acceleration of the flip-flop, the effect increasing with the molecular weight of the halogens. The exception was a fluorinated analog which exhibited slowing of the translocation kinetics. Measurements of the fluorescence ratio of the dye di-4-ANEPPS in lipid vesicles showed significant differences in the adsorption of studied hydrophobic anions. Based on these data, it can be concluded that COSAN and COSAN-F2 were located on the surface of the lipid membrane in the cisoid conformation increasing the dipole potential of the lipid membrane, while other halogenated COSAN analogs were adsorbed in the transoid conformation. Differences in the flip-flop kinetics of COSAN analogs were attributed to variation in the molecular volume of the anions and their orientation on the membrane surface.


Internet-Based Patient Survey on Urolithiasis Treatment and Patient Satisfaction.
We created an Internet-based survey of patients treated for urolithiasis to evaluate for trends in treatment, outcome, and patient satisfaction and to establish internet surveys as a feasible medium for future research of patient urolithiasis treatment experiences. We used the website "kidneystoners.org" to disseminate the online survey, which queried respondents on treatment type, outcome, and satisfaction. Patient satisfaction was correlated with treatment type and outcome. Chi-square and analysis of variance tests were used to compare responses between treatment types. Four hundred forty-three respondents completed the survey. The majority (46%) were treated ureteroscopically, followed by extracorporeal shock wave lithotripsy (SWL, 25%) and percutaneous nephrolithotomy (7%). Other treatments included spontaneous passage (13%), medical expulsive therapy (7%), and home remedies (2%). Sixty-four percent of respondents deemed their treatment "successful," while 36% reported their treatment as either "partially successful" or "unsuccessful." Unsuccessful treatment was more likely for SWL (17%) and home remedies (14%) (p=0.002). Most respondents (52%) reported being either satisfied or very satisfied with their treatment choice. Satisfaction did not vary significantly by treatment type, but was significantly associated with treatment success (mean satisfaction 3.8/5 for "successful" vs 1.9/5 for "unsuccessful" treatment; p<0.0001). Use of the Internet allows rapid gathering of patient information from a large geographic distribution. Our survey is consistent with previous studies in demonstrating an increased use of ureteroscopy to treat both renal and ureteral calculi. In general, patients are satisfied with treatment outcomes despite a large percentage of people reporting needing to have secondary procedures.


Usefulness of serial electrocardiograms for diagnosis of acute myocardial infarction.
The purpose of this study was to determine whether the automated detection of acute myocardial infarction (AMI) by utilizing artificial neural networks was improved by using a previous electrocardiogram (ECG) in addition to the current ECG. A total of 4,691 ECGs were recorded from patients admitted to an emergency department due to suspected AMI. Of these, 902 ECGs, in which diagnoses of AMI were later confirmed, formed the study group, whereas the remaining 3,789 ECGS comprised the control group. For each ECG recorded, a previous ECG of the same patient was selected from the clinical electrocardiographic database. Artificial neural networks were then programmed to detect AMI based on either the current ECG only or on the combination of the previous and the current ECGs. On this basis, 3 assessors--a neural network, an experienced cardiologist, and an intern--separately classified the ECGs of the test group, with and without access to the previous ECG. The detection performance, as measured by the area under the receiver operating characteristic curve, showed an increase for all assessors with access to previous ECGs. The neural network improved from 0.85 to 0.88 (p = 0.02), the cardiologist from 0.79 to 0.81 (p = 0.36), and the intern from 0.71 to 0.78 (p <0.001). Thus, the performance of a neural network, detecting AMI in an ECG, is improved when a previous ECG is used as an additional input.


Revised D-A-CH intake recommendations for folate: how much is needed?
The D-A-CH reference value (D-A-CH arises from the initial letters of the common country identification for the countries Germany (D), Austria (A) and Switzerland (CH)) for folate equivalents had been set at 400 μg/d for adults in the year 2000. By that time, the prevention of cardiovascular diseases through reduction of homocysteine was considered an important target of the reference value. Since that time a number of research papers revealed that in spite of an inverse association between folate-rich diet and chronic diseases, a preventive effect of folic acid intake on cardiovascular events was not supported by randomized controlled trials, and the reduction of plasma homocysteine levels to around 10-12 μmol/l did not reduce the risk for thromboembolic and cardiovascular diseases in persons already affected by these diseases. These results together with the observation that folate intakes below 400 μg/d result in a sufficient folate status justified a review of the current literature and-consequently-a reduction of the reference value to 300 μg/d for adults. This reference value is expressed as dietary folate equivalents that take into account the difference in bioavailability between folic acid and all types of folates in food. The recommendation to take a daily supplement of 400 μg of synthetic folic acid for women who intend to get pregnant and until the end of the first trimester of pregnancy is maintained.


Influence of HLA-DR and -DQ alleles on autoantibody recognition of distinct epitopes within the juxtamembrane domain of the IA-2 autoantigen in type 1 diabetes.
Insulinoma-associated protein 2 (IA-2) is a major target of autoimmunity in type 1 diabetes. When first detected, IA-2-autoantibodies commonly bind epitopes in the juxtamembrane (JM) domain of IA-2 and antibody responses subsequently spread to the tyrosine phosphatase domain. Definition of structures of epitopes in the JM domain, and genetic requirements for autoimmunity to these epitopes, is important for our understanding of initiation and progression of autoimmunity. The aims of this study were to investigate the contribution of individual amino acids in the IA-2 JM domain to antibody binding to these epitopes and the role of HLA genotypes in determining epitope specificity. Regions of the JM domain recognised by autoantibodies were identified by peptide competition and inhibitory effects of alanine substitutions of residues within the JM region. Antibody binding was determined by radioligand binding assays using sera from patients genotyped for HLA-DRB1 and -DQB1 alleles. Patients were categorised into two distinct groups of JM antibody reactivity according to peptide inhibition. Inhibition by substitutions of individual amino acids within the JM domain differed between patients, indicating heterogeneity in epitope recognition. Cluster analysis defined six groups of residues having similar inhibitory effects on antibody binding, with three clusters showing differences in patients affected or unaffected by peptide. One cluster demonstrated significant differences in antibody binding between HLA-DRB1*04 and HLA-DRB1*07 patients and within DRB1*04 individuals; antibody recognition of a second cluster depended on expression of HLA-DQB1*0302. The results identify amino acids contributing to distinct epitopes on IA-2, with both HLA-DR and HLA-DQ alleles influencing epitope specificity.


Wnt signaling regulates mitochondrial physiology and insulin sensitivity.
Mitochondria serve a critical role in physiology and disease. The genetic basis of mitochondrial regulation in mammalian cells has not yet been detailed. We performed a large-scale RNAi screen to systematically identify genes that affect mitochondrial abundance and function. This screen revealed previously unrecognized roles for >150 proteins in mitochondrial regulation. We report that increased Wnt signals are a potent activator of mitochondrial biogenesis and reactive oxygen species (ROS) generation, leading to DNA damage and acceleration of cellular senescence in primary cells. The signaling protein insulin receptor substrate-1 (IRS-1), shown here to be a transcriptional target of Wnt, is induced in this setting. The increased level of IRS-1 drives activation of mitochondrial biogenesis; furthermore, in insulin-responsive cell types, it enhances insulin signaling, raising the possibility that Wnt proteins may be used to modulate glucose homeostasis. Our results identify a key component of the mitochondrial regulatory apparatus with a potentially important link to metabolic and degenerative disorders.


Youth Substance Use Prevention Using Disciplinary Literacy Strategies: A Pilot Study.
Effective health education programs for adolescent substance use prevention need to be developed. However, the evidence supporting the use of specific instructional strategies for youth substance use prevention programs is insufficient. The purpose of this study was to develop and implement a community-based program aiming for smoking, alcohol, and substance use prevention based on multiple disciplinary literacy strategies. A one-group pretest-and-posttest comparison design was used to test the feasibility of the new substance use prevention program. Thirty-two adolescents, most from families with minority and low-socioeconomic status backgrounds, participated. The group met for a 1-hour session once a week for 10 weeks. The program was implemented in a suburban setting in a southeastern state in the United States. Pretests and posttests were administered to assess the program's effectiveness in improving participants' knowledge, skills, and assets regarding substance use. Overall, the participants were satisfied with the program and reported significant growth in their knowledge, skills, assets, and perceptions of benefits. These findings suggest that disciplinary literacy strategies may promote adolescents' perceptions of and knowledge regarding substance use.


Lidar measurements of Raman scattering at ultraviolet wavelength from mineral dust over East Asia.
We developed a novel measurement channel that utilizes Raman scattering from silicon dioxide (SiO2) quartz at an ultraviolet wavelength (361 nm). The excitation of the Raman signals is done at the primary wavelength of 355 nm emitted from a lidar instrument. In combination with Raman signals from scattering from nitrogen molecules, we may infer the mineral-quartz-related backscatter coefficient. This technique thus allows us to identify in a comparably direct way the mineral quartz content in mixed pollution plumes that consist, e.g., of a mix of desert dust and urban pollution. We tested the channel for the complex situation of East Asian pollution. We find good agreement of the inferred mineral-quartz-related backscatter coefficient to results obtained with another mineral quartz channel which was operated at 546 nm (primary emission wavelength at 532 nm), the functionality of which has already been shown for a lidar system in Tsukuba (Japan). The advantage of the novel channel is that it provides a better signal-to-noise ratio because of the shorter measurement wavelength.


Familial clustering of risk factors for cardiovascular disease among first-degree relatives of patients with chronic kidney disease in a sub-Saharan African population.
To determine the prevalence of risk factors for cardiovascular disease (CVD) in first-degree relatives (FDRs) of patients with chronic kidney disease (CKD) in a sub-Saharan African population. This was a cross-sectional survey of 460 subjects (230 FDRs of patients with CKD and 230 healthy controls). Anthropometrics and blood pressures were measured. Spot urine and fasting venous blood samples were obtained for biochemical analysis. The prevalence of hypertension, diabetes mellitus, obesity and dyslipidaemia were significantly higher in FDRs of patients with CKD compared with the controls: 56 (24.3%) vs 29 (12.6%), p = 0.01; 20 (8.7%) vs 6 (2.6%), p = 0.01; 40 (17.4%) vs 24 (10.4%), p = 0.03 and 171 (74.3%) vs 138 (60.0%), p = 0.01, respectively. Hypertension (OR, 1.65), dyslipidaemia (OR, 1.72) and albuminuria (OR, 1.61) were independently associated with being a FDR of patients with CKD. In this sub-Saharan African population, risk factors for CVD were more prevalent in the FDRs of patients with CKD than in healthy controls.


Emerging fabrication techniques for 3D nano-structuring in plasmonics and single molecule studies.
The application of new methods and techniques to fields such as biology and medicine is becoming more and more demanding since the request of detailed information down to single molecules is a scientific necessity and a technical realistic possibility. In this effort a key role is played by emerging fabrication techniques. One of the hardest challenges is to incorporate the third dimension in the design and fabrication of novel devices. Significantly, this means that conventional nano-fabrication methods, intrinsically useful for planar structuring, have to be substituted or complemented with new approaches. In this paper we show how emerging techniques can be used for 3D structuring of noble metals down to nanoscale. In particular, the paper deals with electroless deposition of silver, ion and electron beam induced deposition, focused ion beam milling, and two-photon lithography. We exploited these techniques to fabricate different plasmonics nanolenses, nanoprobes and novel beads for optical tweezers. In the future these devices will be used for the manipulation and chemical investigation of single cells with sensitivity down to a few molecules in label free conditions and native environment. Although this paper is only devoted to nanofabrication, we foresee that the fields of biology and medicine will directly gain substantial advantages from this approach.


Increased risk of Parkinsonism among patients with cirrhosis: a 7-year follow-up study.
Previous studies have suggested that hepatic (toxic-metabolic) encephalopathy, the major complication of cirrhosis, is a neuropsychiatric disorder typically seen in patients with liver dysfunction after exclusion of other known brain disease. This study aims to investigate the risk for parkinsonism during a 7-year follow-up period after a diagnosis of cirrhosis. In total, 1361 patients with cirrhosis and 6805 comparison patients without cirrhosis were included in this study. Each patient was then individually tracked for 7 years from the time of their initial diagnosis of cirrhosis to identify those who developed parkinsonism during the follow-up period. Stratified Cox proportional hazard regressions were conducted to calculate the hazard of parkinsonism for the two groups during the follow-up period, after adjusting for patient's age, monthly income, level of urbanization and geographic location. Of the total 8166 sampled patients, 141 (1.7%) developed parkinsonism during the follow-up period, 48 from the study group (3.5% of the patients with cirrhosis) and 93 from the comparison group (1.4% of patients in the comparison group). Stratified Cox proportional hazard regressions show that the hazard for parkinsonism for patients with cirrhosis was 2.65 times as high (95% confidence interval=1.85-3.80, P<0.001) as the patients in the comparison group over the 7-year follow-up period, after adjusting for patient's age, monthly income, level of urbanization and the geographic location of the community in which the patient resided. We concluded that cirrhosis significantly increased the risk of parkinsonism.


The MprF protein is required for lysinylation of phospholipids in listerial membranes and confers resistance to cationic antimicrobial peptides (CAMPs) on Listeria monocytogenes.
Pathogenic bacteria have to cope with defence mechanisms mediated by adaptive and innate immunity of the host cells. Cationic antimicrobial peptides (CAMPs) represent one of the most effective components of the host innate immune response. Here we establish the function of Lmo1695, a member of the VirR-dependent virulence regulon, recently identified in Listeria monocytogenes. Lmo1695 encodes a membrane protein of 98 kDa with strong homology to the multiple peptide resistance factor (MprF) of Staphylococcus aureus. Like staphylococcal MprF, we found that Lmo1695 is involved in the synthesis of the membrane phospholipid lysylphosphatidylglycerol (L-PG). In addition, Lmo1695 is also essential for lysinylation of diphosphatidylglycerol (DPG), another phospholipid widely distributed in bacterial membranes. A Deltalmo1695 mutant lacking the lysinylated phospholipids was particularly susceptible to CAMPs of human and bacterial origin. The mutant strain infected both epithelial cells and macrophages only poorly and was attenuated for virulence when tested in a mouse model of infection. Lmo1695 is a member of a growing list of survival factors which enable growth of L. monocytogenes in different environments.


A binder-free wet chemical synthesis approach to decorate nanoflowers of bismuth oxide on Ni-foam for fabricating laboratory scale potential pencil-type asymmetric supercapacitor device.
The present study involves the synthesis of a bismuth oxide (Bi2O3) electrode consisting of an arranged nano-platelets for evolving a flower-type surface appearance on nickel-foam (Bi2O3-Ni-F) by a simple, inexpensive, binder-free and one-step chemical bath deposition (CBD) method, popularly known as a wet chemical method. The as-prepared Bi2O3 on Ni-foam, as an electrode material, demonstrates 557 F g-1 specific capacitance (SC, at 1 mA cm-2), of which 85% is retained even after 2000 cycles. With specific power density of 500 kW kg-1, the Bi2O3-Ni-F electrode documents a specific energy density of 80 Wh kg-1. Furthermore, a portable asymmetric supercapacitor device, i.e. a pencil-type cell consisting of Bi2O3-Ni-F as an anode and graphite as a cathode in 6 M KOH aqueous electrolyte solution, confirms 11 Wh kg-1 and 720 kW kg-1 specific energy and specific power densities, respectively. An easy and a simple synthesis approach for manufacturing a portable laboratory scale pencil-type supercapacitor device is a major outcome of this study, which can also be applied for ternary and quaternary metal oxides for recording an enhanced performance. In addition, we presented a demonstration of lighting a light emitting diode (LED) using a home-made pencil-type supercapacitor device which, finally, has confirmed the scaling and technical potentiality of Bi2O3-Ni-F in energy storage devices.


Dietary antioxidants and long-term risk of dementia.
The Rotterdam Study previously found that higher dietary intakes of vitamins E and C related to lower risk of dementia and Alzheimer disease (AD) over 6 years of follow-up. To study consumption of major dietary antioxidants relative to long-term risk of dementia. Population-based prospective cohort study. The Rotterdam Study in the Netherlands. A total of 5395 participants, 55 years and older, who were free of dementia and provided dietary information at study baseline. Incidence of dementia and AD, based on internationally accepted criteria, relative to dietary intake of vitamin E, vitamin C, beta carotene, and flavonoids. During a mean follow-up period of 9.6 years, dementia developed in 465 participants, of whom 365 were diagnosed as having AD. In multivariate models adjusted for age, education, apolipoprotein E epsilon4 genotype, total energy intake, alcohol intake, smoking habits, body mass index, and supplement use, higher intake of vitamin E at study baseline was associated with lower long-term risk of dementia (P = .02 for trend). Compared with participants in the lowest tertile of vitamin E intake, those in the highest tertile were 25% less likely to develop dementia (hazard ratio, 0.75; 95% confidence interval, 0.59-0.95 with adjustment for potential confounders). Dietary intake levels of vitamin C, beta carotene, and flavonoids were not associated with dementia risk after multivariate adjustment (P > .99 for trend for vitamin C and beta carotene and P = .60 for trend for flavonoids). Results were similar when risk for AD was specifically assessed. Higher intake of foods rich in vitamin E may modestly reduce long-term risk of dementia and AD.