The influence of illness‐related variables,personal resources and context‐related factors on real‐life functioning of people with schizophrenia

In people suffering from schizophrenia, major areas of everyday life are impaired, including independent living, productive activities and social relationships. Enhanced understanding of factors that hinder real‐life functioning is vital for treatments to translate into more positive outcomes. The goal of the present study was to identify predictors of real‐life functioning in people with schizophrenia, and to assess their relative contribution. Based on previous literature and clinical experience, several factors were selected and grouped into three categories: illness‐related variables, personal resources and context‐related factors. Some of these variables were never investigated before in relationship with real‐life functioning. In 921 patients with schizophrenia living in the community, we found that variables relevant to the disease, personal resources and social context explain 53.8% of real‐life functioning variance in a structural equation model. Neurocognition exhibited the strongest, though indirect, association with real‐life functioning. Positive symptoms and disorganization, as well as avolition, proved to have significant direct and indirect effects, while depression had no significant association and poor emotional expression was only indirectly and weakly related to real‐life functioning. Availability of a disability pension and access to social and family incentives also showed a significant direct association with functioning. Social cognition, functional capacity, resilience, internalized stigma and engagement with mental health services served as mediators. The observed complex associations among investigated predictors, mediators and real‐life functioning strongly suggest that integrated and personalized programs should be provided as standard treatment to people with schizophrenia.     

GLUT4 Defects in Adipose Tissue Are Early Signs of Metabolic Alterations in Alms1GT/GT,a Mouse Model for Obesity and Insulin Resistance

Dysregulation of signaling pathways in adipose tissue leading to insulin resistance can contribute to the development of obesity-related metabolic disorders. Alström Syndrome, a recessive ciliopathy, caused by mutations in ALMS1, is characterized by progressive metabolic alterations such as childhood obesity, hyperinsulinemia, and type 2 diabetes. Here we investigated the role of Alms1 disruption in AT expansion and insulin responsiveness in a murine model for Alström Syndrome. A gene trap insertion in Alms1 on the insulin sensitive C57BL6/Ei genetic background leads to early hyperinsulinemia and a progressive increase in body weight. At 6 weeks of age, before the onset of the metabolic disease, the mutant mice had enlarged fat depots with hypertrophic adipocytes, but without signs of inflammation. Expression of lipogenic enzymes was increased. Pre-adipocytes isolated from mutant animals demonstrated normal adipogenic differentiation but gave rise to mature adipocytes with reduced insulin-stimulated glucose uptake. Assessment of whole body glucose homeostasis revealed glucose intolerance. Insulin stimulation resulted in proper AKT phosphorylation in adipose tissue. However, the total amount of glucose transporter 4 (SLC4A2) and its translocation to the plasma membrane were reduced in mutant adipose depots compared to wildtype littermates. Alterations in insulin stimulated trafficking of glucose transporter 4 are an early sign of metabolic dysfunction in Alström mutant mice, providing a possible explanation for the reduced glucose uptake and the compensatory hyperinsulinemia. The metabolic signaling deficits either reside downstream or are independent of AKT activation and suggest a role for ALMS1 in GLUT4 trafficking. Alström mutant mice represent an interesting model for the development of metabolic disease in which adipose tissue with a reduced glucose uptake can expand by de novo lipogenesis to an obese state.     

Developmental differences in B cell receptor-induced signal transduction

We have compared early signaling events at various stages of B cell differentiation using established mouse cell lines. Clustering of pre-B cell antigen receptor (BCR) or BCR induced the tyrosine phosphorylation of various proteins in all cells, although the phosphorylation pattern differed. In spite of the pre-BCR-induced tyrosine phosphorylation, we could not detect an intracellular Ca(2+) signal in pre-B cells. However, co-clustering of the pre-BCR with CD19 did induce Ca(2+) mobilization. In contrast to the immature and mature B cells, where the B cell linker protein (BLNK) went through inducible tyrosine phosphorylation upon BCR clustering, we observed a constitutive tyrosine phosphorylation of BLNK in pre-B cell lines. Both BLNK and phospholipase C (PLC)gamma were raft associated in unstimulated pre-B cells, and this could not be enhanced by pre-BCR engagement, suggesting a ligand-independent PLC gamma-mediated signaling. Further results indicate that the cell lines representing the immature stage are more sensitive to BCR-, CD19- and type II receptors binding the Fc part of IgG (Fc gamma RIIb)-mediated signals than mature B cells.     

Combined effects of temperature and salinity on functional responses of haemocytes and survival in air of the clam Ruditapes philippinarum

The combined effects of temperature and salinity on both immune responses and survival in air of the clam, Ruditapes philippinarum, were evaluated for the first time. The animals were kept for 7 days at three differing temperature (5 °C, 15 °C, 30 °C) and salinity values (18 psu, 28 psu, 38 psu), and effects of the resulting 9 experimental conditions on total haemocyte count (THC), Neutral Red uptake (NRU), haemolymph protein concentration, and lysozyme activity in both haemocyte lysate (HL) and cell-free haemolymph (CFH) were evaluated. The survival-in-air test was also performed. Two-way ANOVA analysis revealed that temperature influenced significantly THC and NRU, whereas salinity and temperature/salinity interaction affected NRU only. Temperature and salinity did not influence significantly HL and CFH lysozyme activity, as well as haemolymph total protein content. Survival-in-air test is widely used to evaluate general stress conditions in clams. In the present study, temperature and salinity were shown to influence the resistance to air exposure of R. philippinarum. The highest LT₅₀ (air exposure time resulting in 50% mortality) value was recorded in clams kept at 18 psu and 15 °C, whereas the lowest value was observed in clams kept at 28 psu and 30 °C. Overall, results obtained demonstrated that temperature and salinity can affect some functional responses of haemocytes from R. philippinarum, and suggested a better physiological condition for animals kept at 15 °C temperature and 18 psu salinity.     

Structural determinants of protein translocation in bacteria: conformational flexibility of SecA IRA1 loop region

Bacteria employ the SecA motor protein to push unfolded proteins across the cytoplasmic membrane through the SecY protein‐conducting channel complex. The crystal structure of the SecA–SecY complex shows that the intramolecular regulator of ATPase1 (IRA1) SecA domain, made up of two helices and the loop between them, is partly inserted into the SecY conducting channel, with the loop between the helices as the main functional region. A computational analysis suggested that the entire IRA1 domain is structurally autonomous, and was the basis to synthesize peptide analogs of the SecA IRA1 loop region, to the aim of investigating its conformational preferences. Our study indicates that the loop region populates a predominantly flexible state, even in the presence of structuring agent. This provides indirect evidence that the SecA loop–SecY receptor docking involves loop‐mediated opening of the SecY channel. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd.     

Diarrheagenic Escherichia coli in acute gastroenteritis in infants in North-West Italy

Enteropathogenic Escherichia coli may cause diarrhoea in infancy, but it is not routinely detected and regarded as a major causative agent. The aim of the present study was to estimate the incidence of enteropathogenic E. coli infection and to investigate its epidemiology and pathogenesis from faecal specimens in infants hospitalized for acute gastroenteritis. Between March 2008 and June 2009, faecal samples were collected and examined to recognize diarrhoeal aetiology, especially for E. coli, by cultural identification and multiplex-PCR. E. coli were isolated in 75 of 160 collected samples (46,88%); 10 samples of which (6,3%) had been positively recognised for pathogenic genes. Data showed that the presence of diarrheagenic E. coli infection was 6.3%, but it becomes 5% considering E. coli as a unique agent responsible for diarrhoea. The datum is not statistically meaningful because of the small sample (p>0,05). Bacterial pathogens were also isolated in 60 samples (37,5% of the total collected samples): 15 Salmonella spp., 8 Klebsiella pneumoniae, 9 Klebsiella oxytoca, 11 Citrobacter freundii, 5 Pseudomonas aeruginosa, 2 Serratia spp., 7 Enterobacter cloacae, 1 Shigella spp., 2 Campylobacter spp. Rotavirus was the predominant pathogenic single etiologic agent identified. It was found in 35 samples (21.88% of the overall collected samples), while Adenovirus, serotypes 40 or 41, was isolated in 2 samples (1.3%). Rotavirus infection was found predominantly in winter with respect to autumn. Data provide an interesting epidemiologic survey of enteropathogenic E. coli, which is not usually detected, although it may have potential clinical implications. Abbreviations: CDEC, detaching E. coli; DAEC, diffusely adherent E. coli; EAggEC, enteroaggregative E. coli; EHEC, enterohaemorrhagic E. coli; EIEC, enteroinvasive E. coli; EPEC, enteropathogenic E. coli; ETEC, enterotoxigenic E. coli.     

Contribution of habitat patches to network connectivity: Redundancy and uniqueness of topological indices

In order to increase the efficiency of monitoring and conservation efforts, it is of key importance to develop sound quantitative methods that are able to indicate which key areas and landscape elements play prominent and crucial role in the functioning of habitat mosaics. In particular, network models are being widely used to evaluate the contribution of landscape elements to uphold connectivity and related ecological fluxes. However, monitoring programs and conservation practitioners are overwhelmed by a myriad of network indices without being fully aware of their differences for characterizing the importance of individual habitat patches in fragmented landscapes. We analysed a set of thirteen commonly used graph indices and the forest habitat network of goshawks living in NE Spain in order to (a) evaluate how the patch rank orders derived from these indices differ from each other and (b) identify which indices tend to quantify the same network characteristics and which others are quite unique in addressing topological characteristics that are not considered by the rest. We found that most of the variability in patch rankings can be captured by only three network indices. The largest group of redundant indices corresponded to those that intend to measure the amount of flux received by a given habitat patch. The connector fraction of the integral index of connectivity (IIC) and probability of connectivity (PC) indices and betweenness centrality (BC) stood out as quite unique by focusing on the way habitat patches act as connecting elements between other habitat areas. We discuss which indices can be most beneficial by clearly indicating and differentiating the value of the top patches compared to the others, so that conservation priorities can be established with lower uncertainties. We believe that our results can provide valuable guidelines by facilitating the selection of a few non-redundant and complementary indicators that quantify the important and distinctive roles of habitat patches in maintaining the connectivity of habitat networks.     

Combinatorial analysis and algorithms for quasispecies reconstruction using next-generation sequencing

Background  

Next-generation sequencing (NGS) offers a unique opportunity for high-throughput genomics and has potential to replace Sanger sequencing in many fields, including de-novo sequencing, re-sequencing, meta-genomics, and characterisation of infectious pathogens, such as viral quasispecies. Although methodologies and software for whole genome assembly and genome variation analysis have been developed and refined for NGS data, reconstructing a viral quasispecies using NGS data remains a challenge. This application would be useful for analysing intra-host evolutionary pathways in relation to immune responses and antiretroviral therapy exposures. Here we introduce a set of formulae for the combinatorial analysis of a quasispecies, given a NGS re-sequencing experiment and an algorithm for quasispecies reconstruction. We require that sequenced fragments are aligned against a reference genome, and that the reference genome is partitioned into a set of sliding windows (amplicons). The reconstruction algorithm is based on combinations of multinomial distributions and is designed to minimise the reconstruction of false variants, called in-silico recombinants.     

Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17

In this work, an innovative and non-radioactive functional cAMP assay was validated at the GPR17 receptor. This assay provides a simple and powerful new system to monitor G protein-coupled receptor activity through change in the intracellular cAMP concentration by using a mutant form of Photinus pyralis luciferase into which a cAMP-binding protein moiety has been inserted. Results, expressed as EC₅₀ or IC₅₀ values for agonists and antagonists, respectively, showed a strong correlation with those obtained with [³⁵S]GTPγS binding assay, thus confirming the validity of this approach in the study of new ligands for GPR17. Moreover, this method allowed confirming that GPR17 is coupled with a G_αi.