A fixed combination of probiotics and herbal extracts attenuates intestinal barrier dysfunction from inflammatory stress in an in vitro model using Caco-2 cells.

Background: Inflammatory Bowel Diseases (IBD), are considered a growing global disease, with about ten million people being affected worldwide. Maintenance of intestinal barrier integrity is crucial for preventing IBD onset and exacerbations. Some recent patents regarding oily formulations containing probiotics (WO2010122107A1 and WO2010103374A9) and the use of probiotics for gastrointestinal complaints (US20110110905A1 and US9057112B2) exist, or are pending application. Objective: In this work, we studied the effect of a fixed combination of registered Lactobacillus reuteri and Lactobacillus acidophilus strains and herbal extracts in an in vitro inflammation experimental model. Methods: Caco-2 cell monolayer was exposed to INF-\u3b3+TNF-\u3b1 or to LPS; Trans Epithelial Electrical Resistance (TEER) and paracellular permeability were investigated. ZO-1 and occludin Tight Junctions (TJs) were also investigated by mean of immunofluorescence. Results: Pre-treatment with the fixed combination of probiotics and herbal extracts prevented the inflammation-induced TEER decrease, paracellular permeability increase and TJs translocation. Conclusions: In summary, the fixed combination of probiotics and herbal extracts investigated in this research was found to be an interesting candidate for targeting the re-establishment of intestinal barrier function in IBD conditions

Lactobacillus rhamnosus lowers zebrafish lipid content by changing gut microbiota and host transcription of genes involved in lipid metabolism.

The microbiome plays an important role in lipid metabolism but how the introduction of probiotic communities affects host lipid metabolism is poorly understood. Using a multidisciplinary approach we addressed this knowledge gap using the zebrafish model by coupling high-throughput sequencing with biochemical, molecular and morphological analysis to evaluate the changes in the intestine. Analysis of bacterial 16S libraries revealed that Lactobacillus rhamnosus was able to modulate the gut microbiome of zebrafish larvae, elevating the abundance of Firmicutes sequences and reducing the abundance of Actinobacteria. The gut microbiome changes modulated host lipid processing by inducing transcriptional down-regulation of genes involved in cholesterol and triglycerides metabolism (fit2, agpat4, dgat2, mgll, hnf4α, scap, and cck) concomitantly decreasing total body cholesterol and triglyceride content and increasing fatty acid levels. L. rhamnosus treatment also increased microvilli and enterocyte lengths and decreased lipid droplet size in the intestinal epithelium. These changes resulted in elevated zebrafish larval growth. This integrated system investigation demonstrates probiotic modulation of the gut microbiome, highlights a novel gene network involved in lipid metabolism, provides an insight into how the microbiome regulates molecules involved in lipid metabolism, and reveals a new potential role for L. rhamnosus in the treatment of lipid disorders

Relazioni intercorrenti fra autostima e vittimizzazione da bullismo e cyberbullismo . Risultati di una ricerca condotta tra gli studenti di una scuola secondaria di primo grado

Uno dei fenomeni sociali pi\uf9 diffusamente discussi oggigiorno \ue8 quello del bullismo, comportamento intenzionale di tipo violento e vessatorio nei confronti di soggetti incapaci di difendersi. Tale problematica si correla in maniera interdipendente con i livelli di autostima dei giovanissimi che, a loro volta, hanno ricadute sugli outcomes di benessere e qualit\ue0 della vita in et\ue0 evolutiva. Scopo del presente studio \ue8 quindi indagare i nessi esistenti tra livelli di autostima e bullismo. Materiali e metodi Il Test di valutazione Multidimensionale dell\u2019Autostima (TMA, Bracken), finalizzato alla rilevazione dell\u2019autostima in et\ue0 evolutiva, \ue8 stato somministrato a tutti i 426 alunni di una scuola secondaria di primo grado di Milano. Tale questionario standardizzato consta di 150 items organizzati in sei sottodimensioni (Interpersonale, Competenze, Emotivit\ue0, Successo scolastico, Vita familiare, Vissuto corporeo). I punteggi globali e parziali sono stati correlati con variabili di interesse quali l\u2019essere vittima o perpetratore di bullismo o cyberbullismo ed il tempo trascorso davanti ad un device elettronico. Risultati La percentuale di compilazione del questionario \ue8 stata del 89,4% (n = 381). Le vittime di bullismo sono risultate essere il 32,8% degli intervistati (n = 125; M = 33,8%, F = 31,6%). Il 13,6% ha dichiarato di aver subito atti di cyberbullismo (n = 52; M = 13,3%, F = 14,0%). La percentuale di studenti vittima di cyberbullismo sale al 28,0% tra coloro che hanno subito bullismo e le due condizioni risultano correlate statisticamente (p < 0,01 Mantel-Haenszel). Il 21,0% (n = 80) ha dichiarato inoltre di essere perpetratore di atti di cyberbullismo, con una significativa differenza di genere (M = 25,2%, F = 15,8%; RR 1,6 IC95% 1,05-2,43) e con maggiore frequenza negli alunni delle classi terze (22,4%) e seconde (27,1%) rispetto alle prime (12,7%). La vittimizzazione da cyberbullismo risulta essere fattore predisponente alla messa in atto di simili vessazioni (RR = 2,55; IC95% 1,74-3,76). Il valore di autostima globale tra le vittime di bullismo \ue8 significativamente pi\uf9 basso rispetto alle non vittime (87,5 vs 95,4), con ricadute in tutte le sei dimensioni, in particolare Interpersonale ed Emozionale. Nei ragazzi autori di cyberbullismo si rileva un livello di autostima globale inferiore rispetto ai coetanei non perpetratori (86,7 vs 94,5). Infine, l\u2019essere vittima di cyberbullismo correla positivamente con il tempo trascorso davanti a devices elettronici (p<0,01 x2). Il bullismo pu\uf2 essere considerato una problematica di sanit\ue0 pubblica. \uc8 necessario quindi implementare interventi educativi preventiv

Dna methylation changes in fibromyalgia suggest the role of the immune‐inflammatory response and central sensitization

Fibromyalgia (FM) has been explained as a result of gene‐environment interactions. The present study aims to verify DNA methylation differences in eleven candidate genome regions previously associated to FM, evaluating DNA methylation patterns as potential disease biomarkers. DNA methylation was analyzed through bisulfite sequencing, comparing 42 FM women and their 42 healthy sisters. The associations between the level of methylation in these regions were further explored through a network analysis. Lastly, a logistic regression model investigated the regions potentially associated with FM, when controlling for sociodemographic variables and depressive symptoms. The analysis highlighted significant differences in the GCSAML region methylation between patients and controls. Moreover, seventeen single CpGs, belonging to other genes, were significantly different, however, only one cytosine related to GCSAML survived the correction for multiple comparisons. The network structure of methylation sites was different for each group; GRM2 methylation represented a central node only for FM patients. Logistic regression revealed that depressive symptoms and DNA methylation in the GRM2 region were significantly associated with FM risk. Our study encourages better exploration of GCSAML and GRM2 functions and their possible role in FM affecting immune, inflammatory response, and central sensitization of pain

Conjugated Linoleic Acid: good or bad nutrient

Conjugated linoleic acid (CLA) is a class of 28 positional and geometric isomers of linoleic acid octadecadienoic.Currently, it has been described many benefits related to the supplementation of CLA in animals and humans, as in the treatment of cancer, oxidative stress, in atherosclerosis, in bone formation and composition in obesity, in diabetes and the immune system. However, our results show that, CLA appears to be not a good supplement in patients with cachexia

Ultrastructural and biochemical characterization of mechanically adaptable collagenous structures in the edible sea urchin Paracentrotus lividus

The viscoelastic properties of vertebrate connective tissues rarely undergo significant changes withinphysiological timescales, the only major exception being the reversible destiffening of the mammalianuterine cervix at the end of pregnancy. In contrast to this, the connective tissues of echinoderms (seaurchins, starfish, sea cucumbers, etc.) can switch reversibly between stiff and compliant conditions intimescales of around a second to minutes. Elucidation of the molecular mechanism underlying suchmutability has implications for the zoological, ecological and evolutionary field. Important informationcould also arise for veterinary and biomedical sciences, particularly regarding the pathological plasti-cization or stiffening of connective tissue structures. In the present investigation we analyzed aspectsof the ultrastructure and biochemistry in two representative models, the compass depressor ligamentand the peristomial membrane of the edible sea urchin Paracentrotus lividus, compared in three differentmechanical states. The results provide further evidence that the mechanical adaptability of echinodermconnective tissues does not necessarily imply changes in the collagen fibrils themselves. The higher gly-cosaminoglycan (GAG) content registered in the peristomial membrane with respect to the compassdepressor ligament suggests a diverse role of these molecules in the two mutable collagenous tissues.The possible involvement of GAG in the mutability phenomenon will need further clarification. Duringthe shift from a compliant to a standard condition, significant changes in GAG content were detected onlyin the compass depressor ligament. Similarities in terms of ultrastructure (collagen fibrillar assembling)and biochemistry (two alpha chains) were found between the two models and mammalian collagen.Nevertheless, differences in collagen immunoreactivity, alpha chain migration on SDS-PAGE and BLASTalignment highlighted the uniqueness of sea urchin collagen with respect to mammalian collagen

MGMT methylation in diffuse large B-cell lymphoma: validation of quantitative methylation-specific PCR and comparison with MGMT protein expression.

AIMS: (1) To validate a quantitative real time methylation specific PCR assay (MethyLight) for the detection of O6-methylguanine-DNA methyltransferase (MGMT) gene methylation status (MS) in diffuse large B-cell lymphoma (DLBCL). (2) To determine the immunohistochemical (IHC) expression of the MGMT protein and correlate it with MS. Both IHC and MethyLight results were compared with patient's outcome. METHODS: 71 patients with primary nodal DLBCL were studied. MGMT immunoreactivity was detected using a specific monoclonal antibody. The MS of MGMT gene was analysed in 52/71 DLBCL using MethyLight. A selected subset of 40 DLBCL was also analysed using qualitative methylation-specific PCR (MSP). Statistical analysis of overall survival (OS), lymphoma-specific survival (LSS) and progression free survival (PFS) was performed according to IHC and MS results. RESULTS: 19/71 DLBCLs (27%) were MGMT-negative at IHC; all were analysed, together with 33/52 MGMT-positive DLBCLs. MethyLight showed a better performance than MSP. There was a good correlation between the presence of MGMT expression and the unmethylated status; the absence of IHC expression was poorly correlated with the presence of methylation. Better OS, LSS and PFS was found in DLBCLs with MGMT gene methylation. DLBCLs not expressing MGMT at IHC showed a longer PFS. CONCLUSIONS: The quantitative real-time methylation-specific PCR assay for the detection of MGMT gene hypermethylation has been validated for the first time in DLBCL. Immunohistochemistry seems to represent an useful preliminary test to identify unmethylated cases; MS analysis may be performed in non-immunoreactive cases to identify truly methylated DLBCLs, which bear a better prognosis