Transgenic Rescue of the LARGEmyd Mouse: A LARGE Therapeutic Window?

LARGE is a glycosyltransferase involved in glycosylation of α-dystroglycan (α-DG). Absence of this protein in the LARGEmyd mouse results in α-DG hypoglycosylation, and is associated with central nervous system abnormalities and progressive muscular dystrophy. Up-regulation of LARGE has previously been proposed as a therapy for the secondary dystroglycanopathies: overexpression in cells compensates for defects in multiple dystroglycanopathy genes. Counterintuitively, LARGE overexpression in an FKRP-deficient mouse exacerbates pathology, suggesting that modulation of α-DG glycosylation requires further investigation. Here we demonstrate that transgenic expression of human LARGE (LARGE-LV5) in the LARGEmyd mouse restores α-DG glycosylation (with marked hyperglycosylation in muscle) and that this corrects both the muscle pathology and brain architecture. By quantitative analyses of LARGE transcripts we also here show that levels of transgenic and endogenous LARGE in the brains of transgenic animals are comparable, but that the transgene is markedly overexpressed in heart and particularly skeletal muscle (20–100 fold over endogenous). Our data suggest LARGE overexpression may only be deleterious under a forced regenerative context, such as that resulting from a reduction in FKRP: in the absence of such a defect we show that systemic expression of LARGE can indeed act therapeutically, and that even dramatic LARGE overexpression is well-tolerated in heart and skeletal muscle. Moreover, correction of LARGEmyd brain pathology with only moderate, near-physiological LARGE expression suggests a generous therapeutic window

Virtual Reality Laboratories in Engineering Blended Learning Environments: Challenges and Opportunities

A great number of educational institutions worldwide have had their activities partially or fully interrupted following the outbreak of the COVID-19 pandemic. Consequently, universities have had to take the necessary steps in order to adapt their teaching, including laboratory workshops, to a fully online or mixed mode of delivery while maintaining their academic standards and providing a high-quality student experience. This transition has required, among other efforts, adequate investments in tools, accessibility, content development, and competences as well as appropriate training for both the teaching and administrative staff. In such a complex scenario, Virtual Reality Laboratories (VRLabs), which in the past already proved themselves to be efficient tools supporting the traditional practical activities, could well represent a valid alternative in the hybrid didactic mode of the contemporary educational landscape, rethinking the educational proposal in light of the indications coming from the scientific literature in the pedagogical field. In this context, the present work carries out a critical review of the existent virtual labs developed in the Engineering departments in the last ten years (2010-2020) and includes a pre-pandemic experience of a VRLab tool-StreamFlowVR-within the Hydraulics course of Basilicata University, Italy. This analysis is aimed at highlighting how ready VRLabs are to be exploited not only in emergency but also in ordinary situations, together with valorising an interdisciplinary dialogue between the pedagogical and technological viewpoints, in order to progressively foster a high-quality and evidence-based educational experience

Identification of na\uefve HCV-1 patients with chronic hepatitis who may benefit from dual therapy with peg-interferon and ribavirin

Background & Aims The pool of HCV genotype 1 patients likely to be cured by peg-interferon and ribavirin remains to be quantified. Methods In 1045 patients treated with peg-interferon and ribavirin, two therapeutic strategies were confronted: the first one evaluated only baseline variables associated with sustained virological response (SVR), and the second one included the rapid virologic response (RVR) in addition to baseline predictors. An 80% SVR rate was the threshold to retain a strategy as clinically relevant. Results Overall, 414 patients (39.6%) attained SVR. In the first strategy, the hierarchy of features independently associated with SVR was IL28B CC genotype (OR 5.082; CI 3.637-7.101), low (80% SVR threshold. Only 26.6% of no-RVR patients attained SVR. Among the 255 RVR patients, the likelihood of SVR was 61.8% in those with unfavorable predictors, 80% in the presence of a single predictor, and 100% when both predictors were present. By using this model, 200 patients (19.1%) were predicted to have an 80% chance of being cured with dual therapy. Conclusions A consistent subset of na\uefve HCV-1 patients, identified by some baseline characteristics and RVR, may benefit from dual treatment with peg-interferon and ribavirin. \ua9 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved

Praja1 E3 ubiquitin ligase promotes skeletal myogenesis through degradation of EZH2 upon p38α activation

Polycomb proteins are critical chromatin modifiers that regulate stem cell differentiation via transcriptional repression. In skeletal muscle progenitors Enhancer of zeste homologue 2 (EZH2), the catalytic subunit of Polycomb Repressive Complex 2 (PRC2), contributes to maintain the chromatin of muscle genes in a repressive conformation, whereas its down-regulation allows the progression through the myogenic programme. Here, we show that p38α kinase promotes EZH2 degradation in differentiating muscle cells through phosphorylation of threonine 372. Biochemical and genetic evidence demonstrates that the MYOD-induced E3 ubiquitin ligase Praja1 (PJA1) is involved in regulating EZH2 levels upon p38α activation. EZH2 premature degradation in proliferating myoblasts is prevented by low levels of PJA1, its cytoplasmic localization and the lower activity towards unphosphorylated EZH2. Our results indicate that signal-dependent degradation of EZH2 is a prerequisite for satellite cells differentiation and identify PJA1 as a new player in the epigenetic control of muscle gene expression

Micromechanical Properties of Injection-Molded Starch–Wood Particle Composites

The micromechanical properties of injection molded starch–wood particle composites were investigated as a function of particle content and humidity conditions. The composite materials were characterized by scanning electron microscopy and X-ray diffraction methods. The microhardness of the composites was shown to increase notably with the concentration of the wood particles. In addition,creep behavior under the indenter and temperature dependence were evaluated in terms of the independent contribution of the starch matrix and the wood microparticles to the hardness value. The influence of drying time on the density and weight uptake of the injection-molded composites was highlighted. The results revealed the role of the mechanism of water evaporation, showing that the dependence of water uptake and temperature was greater for the starch–wood composites than for the pure starch sample. Experiments performed during the drying process at 70°C indicated that the wood in the starch composites did not prevent water loss from the samples.Peer reviewe

Pilot study: a descriptive-retrospective analysis of SARS-CoV-2 variants distribution and phylogenesis in the Phlegraean area

COVID-19 disease, caused by SARS-CoV-2 virus, marked the pandemic era, opening the way to next-generation sequencing in the viral diagnostic field. SARS-CoV-2 viral genome sequencing makes it possible to identify mutations in the virus and to track the diffusion of these variants in specific geographic area and in time. Variant sequences help understand how the virus spreads and how it can be contained, as well as for developing more effective vaccines and therapies. Indeed, monitoring the evolution of a virus allows us to quickly detect the potential selection of a super mutation, which can make a virus even more contagious and dangerous in terms of human health consequences. In light of this, in our pilot study, we decided to profile the SARS-CoV-2 genome, recruiting 38 patients divided according to age, sex, vaccination status and symptoms, ascertaining their positivity to the virus. Specific strains of SARS-CoV-2 have been identified and effective through next-generation sequencing. This analysis made it possible to obtain information on the variants of the virus and their spread in the Campania region of the Phlegraean area, in the municipalities of Bacoli, Pozzuoli and Monte di Procida from December 2021 to February 2023 and on the effect of long-term measures COVID-19 in our sample. The advantage of using NGS in diagnosis is the introduction of tests on many genes in a relatively short time and at relatively low costs, with a consequent increase in a precise molecular diagnosis and helps to identify ad personam therapies