A multi-element psychosocial intervention for early psychosis (GET UP PIANO TRIAL) conducted in a catchment area of 10 million inhabitants: study protocol for a pragmatic cluster randomized controlled trial

Multi-element interventions for first-episode psychosis (FEP) are promising, but have mostly been conducted in non-epidemiologically representative samples, thereby raising the risk of underestimating the complexities involved in treating FEP in 'real-world' services

Deciphering PDZ mediated protein complexes by Affinity Chromatography and Mass Spectrometry

Small modular binding domains mediate protein-protein interactions by conferring specificity in multiprotein complex formation. The variability of PDZ (PSD-95/Dlg/ZO-1) domains primary sequences and the structural adaptability of their fold results in different binding modalities. PDZs generally bind to the target carboxyl terminus, but they can also bind to internal sequences of other PDZ or other interaction modules. The eight PDZ domains of the protein PATJ (Protein Associated to Tight Junctions) have been assayed with protein extracts by affinity chromatography followed by mass spectrometry in order to find their protein interactors. Fifteen different proteins were indentified and the interactions of some of them with the PDZ domains of PATJ were further investigate

Proteins from Tuber magnatum Pico fruiting bodies naturally grown in different areas of Italy

Background: A number of Tuber species are ecologically important. The fruiting bodies of some of these also have value as a cooking ingredient due to the fact that they possess exceptional flavor and aromatic properties. In particular, T. magnatum fruiting bodies (commonly known as truffles), are greatly appreciated by consumers. These grow naturally in some parts of Italy. However, the quality of these fruiting bodies varies significantly depending on the area of origin due to differences in environmental growth conditions. It is therefore useful to be able to characterize them. A suitable method to reach this goal is to identify proteins which occur in the fruiting bodies that are specific to each area of origin. In this work protein profiles are described for samples coming from different areas and collected in two successive years. To our knowledge this is the first time that proteins of T. magnatum have been thoroughly examined.Results: Using two dimensional electrophoresis, reproducible quantitative differences in the protein patterns (total 600 spots) of samples from different parts of Italy (accession areas) were revealed by bioinformatic analysis. 60 spots were chosen for further analysis, out of which 17 could probably be used to distinguish a sample grown in one area from a sample grown in another area. Mass spectrometry (MS) protein analysis of these seventeen spots allowed the identification of 17 proteins of T. magnatum.Conclusions: The results indicate that proteomic analysis is a suitable method for characterizing those differences occurring in samples and induced by the different environmental conditions present in the various Italian areas where T. magnatum can grow. The positive protein identification by MS analysis has proved that this method can be applied with success even in a species whose genome, at the moment, has not been sequenced. © 2013 Vita et al.; licensee BioMed Central Ltd

Relationship between Type A spermatozoa motility in the ejaculate of infertile treated men and the incidence of pregnancy achieved with artificial insemination

Objective: Aim of this study was to evaluate the relation between Type A spermatozoa motility in ejaculate of treated infertile men and incidence of pregnancy achieved with artificial insemination. Materials and methods. 72 infertile males from couples with only male infertility, were included in the study. Seminal fluid was assayed according to the World Health Organization guidelines and semen was prepared through gradient density for insemination. Couples were followed to evaluate pregnancy occurrence. Four categories of Type A spermatozoa were identified: first = 0-10%, second = 11-30%, third = 31-50% and fourth = 51-100%. Data were analyzed using the logistic regression model to estimate the relative risk (RR). Results. Pregnancy rates in the four Type A spermatozoa categories were: 11.8% (SE = 7.8), 45.0% (SE = 11.1), 26.7% (SE = 11.4) and 30.0% (SE = 10.3). Using the first Type A spermatozoa category as reference, the second category was significantly different in pregnancy incidence (RR = 6.14, p-value = 0.0385), but no difference emerged in the third and fourth category (RR = 2.7, p-value = 0.2924 and RR = 3.2, p-value = 0.1931). Conclusions. This study shows that having Type A spermatozoa percentage lower than 10% is an unfavorable factor, whereas the highest number of pregnancies occurred with spermatozoa in the 11-30% category. Further percentage increase of Type A spermatozoa did not improve the pregnancy rate. Notwithstanding the little sample size, these data suggest that a significant relation exists between Type A spermatozoa in the ejaculate and pregnancy rate

Channel-interacting PDZ protein “CIPP” interacts with proteins involved in cytoskeletal dynamics

International audienceNeuronal CIPP is a multivalent PDZ protein that interacts with specific channels and receptors, highly expressed in the brain. It is composed of four PDZ domains that behave as a scaffold to clusterize functionally connected proteins. In this study, we selected a set of potential CIPP interactors that are directly or indirectly involved in mechanisms of cytoskeletal remodeling and membrane protrusions formation. For some of these, we first proved the direct binding to specific CIPP PDZ domains considered as autonomous elements, and then confirmed the interaction with the whole protein. In particular, the small G-protein effector IRSp53 (insulin receptor tyrosine kinase substrate protein p53) specifically interacts with the second PDZ domain of CIPP and, when co-transfected in mammalian cultured cells with a tagged full-length CIPP, it induces a marked reorganization of CIPP cytoplasmic localization. Large punctate structures are generated as a consequence of CIPP binding to IRSp53 carboxy-terminus. Analysis of the puncta nature, using various endocytic markers, revealed that they are not related to cytoplasmic vesicles, rather represent multi-protein assemblies, where CIPP can tether other potential interactors

Vascular Smooth Muscle Cells activation revealed by quantitative phosphoproteomics analysis

Vascular smooth-muscle cells (VSMCs) are the main components of the artery medial layer and if activated by growth factors (such as PDGF-BB) as a consequence of vessel injuries, acquire the ability to proliferate and migrate contributing to the formation of neointima. In the early times of VSMC stimulation a cascade of kinases and phosphatases initiates phospho-events that are decisive for VSMC activation, which terminates with a reorganization of the cell structure. In this work, a shotgun proteomics approach aimed at disclosing factors involved in these phenotypically imperceptible, but significant events has been applied. A SILAC approach in a multi-strategy combined method for phosphopeptides enrichment was used, in order to gain insights into the phosphomodulation of VSMC proteome stimulated by PDGF-BB.We performed a quantitative SILAC phosphoproteome analysis on primary VSMC cells which allowed the identification of 1300 phosphopeptides among which 47 resulted novel phosphosites. Moreover, in VSMC signaling events, some important factors involved in cytoskeleton remodeling, focal adhesions, gap junction assembly and cell activation have been found differentially phosphorylated, highlighting their pivotal role in early VSMC reorganization and suggesting a novel starting point to assess the precise actors of VSMC activation and their roles

ProForma: A Standard Proteoform Notation

International audienceThe Consortium for Top-Down Proteomics (CTDP) proposes a standardized notation, ProForma, for writing the sequence of fully characterized proteoforms. ProForma provides a means to communicate any proteoform by writing the amino acid sequence using standard one-letter notation and specifying modifications or unidentified mass shifts within brackets following certain amino acids. The notation is unambiguous, human-readable, and can easily be parsed and written by bioinformatic tools. This system uses seven rules and supports a wide range of possible use cases, ensuring compatibility and reproducibility of proteoform annotations. Standardizing proteoform sequences will simplify storage, comparison, and reanalysis of proteomic studies, and the Consortium welcomes input and contributions from the research community on the continued design and maintenance of this standard

Extracellular vesicles from neural stem cells transfer IFN-γ via Ifngr1 to activate Stat1 signaling in target cells

The idea that stem cell therapies work only via cell replacement is challenged by the observation of consistent intercellular molecule exchange between the graft and the host. Here we defined a mechanism of cellular signaling by which neural stem/precursor cells (NPCs) communicate with the microenvironment via extracellular vesicles (EVs), and we elucidated its molecular signature and function. We observed cytokine-regulated pathways that sort proteins and mRNAs into EVs. We described induction of interferon gamma (IFN-γ) pathway in NPCs exposed to proinflammatory cytokines that is mirrored in EVs. We showed that IFN-γ bound to EVs through Ifngr1 activates Stat1 in target cells. Finally, we demonstrated that endogenous Stat1 and Ifngr1 in target cells are indispensable to sustain the activation of Stat1 signaling by EV-associated IFN-γ/Ifngr1 complexes. Our study identifies a mechanism of cellular signaling regulated by EV-associated IFN-γ/Ifngr1 complexes, which grafted stem cells may use to communicate with the host immune system