Skeletal muscle myopenia in mice model of bile duct ligation and carbon tetrachloride-induced liver cirrhosis

Skeletal muscle myopathy is universal in cirrhotic patients, however, little is known about the main mechanisms involved. The study aims to investigate skeletal muscle morphological, histological, and functional modifications in experimental models of cirrhosis and the principal molecular pathways responsible for skeletal muscle myopathy. Cirrhosis was induced by bile duct ligation (BDL) and carbon tetrachloride (CCl4) administration in mice. Control animals (CTR) underwent bile duct exposure or vehicle administration only. At sacrifice, peripheral muscles were dissected and weighed. Contractile properties of extensor digitorum longus (EDL) were studied in vitro. Muscle samples were used for histological and molecular analysis. Quadriceps muscle histology revealed a significant reduction in cross-sectional area of muscle and muscle fibers in cirrhotic mice with respect to CTR. Kinetic properties of EDL in both BDL and CCl4 were reduced with respect to CTR; BDL mice also showed a reduction in muscle force and a decrease in the resistance to fatigue. Increase in myostatin expression associated with a decrease in AKT-mTOR expressions was observed in BDL mice, together with an increase in LC3 protein levels. Upregulation of the proinflammatory citochines TNF-a and IL6 and an increased expression of NF-kB and MuRF-1 were observed in CCl4 mice. In conclusion, skeletal muscle myopenia was present in experimental models of BDL and CCl4-induced cirrhosis. Moreover, reduction in protein synthesis and activation of protein degradation were the main mechanisms responsible for myopenia in BDL mice, while activation of ubiquitin-pathway through inflammatory cytokines seems to be the main potential mechanism involved in CCl4 mice

A novel pathogenic BRCA1 splicing variant produces partial intron retention in the mature messenger RNA

About 10% of all breast cancers arise from hereditary mutations that increase the risk of breast and ovarian cancers; and about 25% of these are associated with the BRCA1 or BRCA2 genes. The identification of BRCA1/BRCA2 mutations can enable physicians to better tailor the clinical management of patients; and to initiate preventive measures in healthy carriers. The pathophysiological significance of newly identified variants poses challenges for genetic counseling. We characterized a new BRCA1 variant discovered in a breast cancer patient during BRCA1/2 screening by next-generation sequencing. Bioinformatic predictions; indicating that the variant is probably pathogenetic; were verified using retro-transcription of the patient's RNA followed by PCR amplifications performed on the resulting cDNA. The variant causes the loss of a canonic donor splice site at position +2 in BRCA1 intron 21; and consequently the partial retention of 156 bp of intron 21 in the patient's transcript; which demonstrates that this novel BRCA1 mutation plays a pathogenetic role in breast cancer. These findings enabled us to initiate appropriate counseling and to tailor the clinical management of this family. Lastly; these data reinforce the importance of studying the effects of sequence variants at the RNA level to verify their potential role in disease onset

Gallbladder stem/progenitor cells are able to repopulate and rescue the damaged liver in a experimental mouse model of liver cirrhosis

[No abstract available]The human biliary tree contains stem cells within peribiliary glands (Cardinale et al., 2011). Human Gallbladder contains stem/progenitors (hGSCs) located in mucosal crypts (Carpino et al., 2012). Our aim was to evaluate the capability of hGSCs to repopulate and rescue the damaged liver in a model of liver cirrhosis. hGSCs were selected for Epithelial Cell Adhesion Molecule. Cirrhosis was induced by intraperitoneal injection, twice a week, of carbon tetrachloride for 7 weeks. hGSCs were injected into the liver via the spleen of normal or cirrhotic SCID mice. As controls, cirrhotic SCID mice were injected only with the medium or with mature human hepatocytes. 2 months after the injection, the necrotic areas were lower in mice treated with hGSCs in comparison with controls. In hGSCs-injected cirrhotic mice, the expression of the human antigens indicated that ≈10% of the host hepatocyte mass was represented by in vivo differentiated human hepatocytes derived from injected hGSCs. This value was significantly higher with respect to mice injected with human adult hepatocytes. The presence of human cells in murine livers was confirmed by in situ hybridization for human chromosomes. hGSCs injection dictated an improvement of serum liver biochemistry with a significant reduction of transaminases and an improvement of synthetic functions. In this model, hGSC engraftment and differentiation determinate the improvement of histological liver damage and serum liver biochemistry. These data could open future perspectives for a role of gallbladder as a source of stem cells for liver regenerative medicine

Coffee prevents fatty liver disease induced by a high-fat diet by modulating pathways of the gut-liver axis

Coffee consumption is inversely associated with the risk of non-alcoholic fatty liver disease (NAFLD). A gap in the literature still exists concerning the intestinal mechanisms that are involved in the protective effect of coffee consumption towards NAFLD. In this study, twenty-four C57BL/6J mice were divided into three groups each receiving a standard diet, a high-fat diet (HFD) or an HFD plus decaffeinated coffee (HFD+COFFEE) for 12 weeks. Coffee supplementation reduced HFD-induced liver macrovesicular steatosis (P\ua0<\ua00\ub701) and serum cholesterol (P\ua0<\ua00\ub7001), alanine aminotransferase and glucose (P\ua0<\ua00\ub705). Accordingly, liver PPAR- \u3b1 (P\ua0<\ua00\ub705) and acyl-CoA oxidase-1 (P\ua0<\ua00\ub705) as well as duodenal ATP-binding cassette (ABC) subfamily A1 (ABCA1) and subfamily G1 (ABCG1) (P\ua0<\ua00\ub705) mRNA expressions increased with coffee consumption. Compared with HFD animals, HFD+COFFEE mice had more undigested lipids in the caecal content and higher free fatty acid receptor-1 mRNA expression in the duodenum and colon. Furthermore, they showed an up-regulation of duodenal and colonic zonulin-1 (P\ua0<\ua00\ub705), duodenal claudin (P\ua0<\ua00\ub705) and duodenal peptide YY (P\ua0<\ua00\ub705) mRNA as well as a higher abundance of Alcaligenaceae in the faeces (P\ua0<\ua00\ub705). HFD+COFFEE mice had an energy intake comparable with HFD-fed mice but starting from the eighth intervention week they gained significantly less weight over time. Data altogether showed that coffee supplementation prevented HFD-induced NAFLD in mice by reducing hepatic fat deposition and metabolic derangement through modification of pathways underpinning liver fat oxidation, intestinal cholesterol efflux, energy metabolism and gut permeability. The hepatic and metabolic benefits induced by coffee were accompanied by changes in the gut microbiota

Celiac disease-associated Neisseria flavescens decreases mitochondrial respiration in CaCo-2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial-induced cellular imbalance

: We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo-2 cells. We also found that vesicular trafficking was delayed after the CD-immunogenic P31-43 gliadin peptide-entered CaCo-2 cells and that Lactobacillus paracasei CBA L74 (L.&nbsp;paracasei-CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD-N.&nbsp;flavescens-infected CaCo-2 cells and if any alteration could be mitigated by pretreating cells with L.&nbsp;paracasei-CBA supernatant, despite the presence of P31-43. We measured CaCo-2 bioenergetics by an extracellular flux analyser, N.&nbsp;flavescens and P31-43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD-N. flavescens colocalised more than control N.&nbsp;flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31-43 increased colocalisation of N.&nbsp;flavescens with early vesicles. Mitochondrial respiration was lower (P&nbsp;&lt;&nbsp;.05) in CD-N.&nbsp;flavescens-infected cells versus not-treated CaCo-2 cells, whereas pretreatment with L.&nbsp;paracasei-CBA reduced CD-N.&nbsp;flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD-N.&nbsp;flavescens induces metabolic imbalance in CaCo-2 cells, and the L.&nbsp;paracasei-CBA probiotic could be used to correct CD-associated dysbiosis

The molecular analysis of BRCA1 and BRCA2: Next-generation sequencing supersedes conventional approaches

Abstract Background Accurate and sensitive detection of BRCA 1/2 germ-line mutations is crucial for the clinical management of women affected by breast cancer, for prevention and, notably, also for the identification of at-risk healthy relatives. The most widely used methods for BRCA1 / 2 molecular analysis are Sanger sequencing, and denaturing high performance liquid chromatography (dHPLC) followed by the Sanger method. However, recent findings suggest that next-generation sequencing (NGS)-based approaches may be an efficient tool for diagnostic purposes. In this context, we evaluated the effectiveness of NGS for BRCA gene analysis compared with dHPLC/Sanger sequencing. Methods Seventy women were screened for BRCA1/2 mutations by both dHPLC/Sanger sequencing and NGS, and the data were analyzed using a bioinformatic pipeline. Results Sequence data analysis showed that NGS is more sensitive in detecting BRCA 1/2 variants than the conventional procedure, namely, dHPLC/Sanger. Conclusion Next-generation sequencing is more sensitive, faster, easier to use and less expensive than the conventional Sanger method. Consequently, it is a reliable procedure for the routine molecular screening of the BRCA 1/2 genes

Electronic cigarette use as an aid to quit smoking in the representative Italian population PASSI survey

This study explored electronic cigarette (e-cigarette) use as an aid to quit smoking and compared abstinence rates for different quitting methods in a representative sample of the Italian population. In the 2014–2015 PASSI survey, the ongoing Italian behavioural risk factor surveillance system, 6112 adults who smoked and made at least one quit attempt in the previous 12 months, were categorized into three groups according to the method used in their most recent quit attempt: e-cigarette only, no aid, other quitting methods (medications; programmes delivered in smoking cessation services; other unspecified methods). The primary outcome was self-reported abstinence for a period ≥ 6 months, adjusted for potential confounders. Eleven percent used e-cigarettes only, 86% no aid, 3% other quitting methods. Smoking abstinence was reported among 9% of those using no aid; 8% of e-cigarette users; 15% of those using other methods. No significant differences in abstinence were observed for e-cigarette users compared with those reporting no aid (adjusted Prevalence Ratio [aPR] = 0.81; 95%Confidence Interval (CI) = 0.58–1.14). Changing the reference group to e-cigarette users, those using other quitting methods were significantly more likely to report abstinence than e-cigarette users (aPR = 1.76; 95%CI = 1.07–2.88). One out of ten smokers who attempted to quit in 2014–2015 in Italy used e-cigarettes. E-cigarettes users were as likely to report abstinence as those using no aid, but were less likely to report abstinence than users of established quitting methods. Further studies are needed to understand the relationship between e-cigarette types used to quit and abstinence rates

ORGANIC, METALLORGANIC AND BIOLOGICAL MATERIALOBTAINED FROM ROCK

L' invenzione riguarda metodi d'ottenimento da campioni di roccia di varia natura di materiale organico, metallorganico o di origine biologica, e i materiali stessi così ottenuti. L'invenzione riguarda altresì il loro uso in apparecchiature che sfruttino le loro proprietà fisiche, fisico-chimiche e metodi fluorimetrici d'identificazione di tali materiali. L'uso delle MICRO/NANO struttureMAGNETICHE oggetto dell'invenzione (o di loro parti componenti) come catalizzatori di reazioni chimiche e/o biochimiche mirate all'idrolisi, alla formazione o alla modificazione di legami estere, peptidico o glicosidico

Organic, metallorganic and biological material obtained from rock

The present invention concerns methods for obtaining from samples of rock of different natures, specifically chondrites, organic, metallorganic material or material of biological origin, and the materials themselves so obtained. In addition, the invention concerns their use in instrumentations exploiting their physical and physico-chemical properties, and fluorimetric methods to reveal such materials