In-House Validation and Comparison of Two Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>) Taxon-Specific Real-Time PCR Methods for GMO Quantification Supported by Droplet Digital PCR

Common wheat is one of the most important staple food crops worldwide. However, unlike other important staple crops such as maize or soybean, genetically modified (GM) wheat is not yet present in the global food market. Nonetheless, in the recent past, the adventitious presence of GM glyphosate-tolerant volunteers was reported in open wheat fields in the USA. The European Union Reference Laboratory for GM Food and Feed (EURL-GMFF) was therefore called to develop a strategy to detect such unauthorised GM wheat in wheat samples by using both taxon-specific and screening tests. Two candidate common wheat taxon-specific real-time PCR methods were suggested, one targeting ssII-D gene coding for starch synthase and the other targeting waxy-D1 gene, coding for granule-bound starch synthase. In the present study, the two above-mentioned real-time PCR taxon-specific methods were in-house verified and compared, proposing droplet digital PCR (ddPCR) as a new tool for supporting the application of the European Network of GMO Laboratories (ENGL) established method performance criteria. Preliminary performance data of waxy-D1 and ssII-D methods in ddPCR format are shown too to give a contribution to the bridging process from the consolidated to the emerging quantitative PCR methodology.     

Perspectives on hiPSC-Derived Muscle Cells as Drug Discovery Models for Muscular Dystrophies

Muscular dystrophies are a heterogeneous group of inherited diseases characterized by the progressive degeneration and weakness of skeletal muscles, leading to disability and, often, premature death. To date, no effective therapies are available to halt or reverse the pathogenic process, and meaningful treatments are urgently needed. From this perspective, it is particularly important to establish reliable in vitro models of human muscle that allow the recapitulation of disease features as well as the screening of genetic and pharmacological therapies. We herein review and discuss advances in the development of in vitro muscle models obtained from human induced pluripotent stem cells, which appear to be capable of reproducing the lack of myofiber proteins as well as other specific pathological hallmarks, such as inflammation, fibrosis, and reduced muscle regenerative potential. In addition, these platforms have been used to assess genetic correction strategies such as gene silencing, gene transfer and genome editing with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), as well as to evaluate novel small molecules aimed at ameliorating muscle degeneration. Furthermore, we discuss the challenges related to in vitro drug testing and provide a critical view of potential therapeutic developments to foster the future clinical translation of preclinical muscular dystrophy studies.     

Polybetaines in Biomedical Applications

Polybetaines, that have moieties bearing both cationic (quaternary ammonium group) and anionic groups (carboxylate, sulfonate, phosphate/phosphinate/phosphonate groups) situated in the same structural unit represent an important class of smart polymers with unique and specific properties, belonging to the family of zwitterionic materials. According to the anionic groups, polybetaines can be divided into three major classes: poly(carboxybetaines), poly(sulfobetaines) and poly(phosphobetaines). The structural diversity of polybetaines and their special properties such as, antifouling, antimicrobial, strong hydration properties and good biocompatibility lead to their use in nanotechnology, biological and medical fields, water remediation, hydrometallurgy and the oil industry. In this review we aimed to highlight the recent developments achieved in the field of biomedical applications of polybetaines such as: antifouling, antimicrobial and implant coatings, wound healing and drug delivery systems.     

Exploring the Role of Skeletal Muscle in Insulin Resistance: Lessons from Cultured Cells to Animal Models

Skeletal muscle is essential to maintain vital functions such as movement, breathing, and thermogenesis, and it is now recognized as an endocrine organ. Muscles release factors named myokines, which can regulate several physiological processes. Moreover, skeletal muscle is particularly important in maintaining body homeostasis, since it is responsible for more than 75% of all insulin-mediated glucose disposal. Alterations of skeletal muscle differentiation and function, with subsequent dysfunctional expression and secretion of myokines, play a key role in the pathogenesis of obesity, type 2 diabetes, and other metabolic diseases, finally leading to cardiometabolic complications. Hence, a deeper understanding of the molecular mechanisms regulating skeletal muscle function related to energy metabolism is critical for novel strategies to treat and prevent insulin resistance and its cardiometabolic complications. This review will be focused on both cellular and animal models currently available for exploring skeletal muscle metabolism and endocrine function.     

Procedures for fault location and isolation to solve protection selectivity problems in MV distribution networks with dispersed generation

Present planning and operation criteria used for MV passive radial distribution networks are, in general, not suitable to cope with the presence of a significant distributed generation (DG) capacity. In fact, many technical problems are still to be solved in order to increase DG penetration and to provide, at the same time, high service quality levels to customers.     

High Glucose Exposure Impairs L-Cell Differentiation in Intestinal Organoids: Molecular Mechanisms and Clinical Implications

Intestinal organoids are used to analyze the differentiation of enteroendocrine cells (EECs) and to manipulate their density for treating type 2 diabetes. EEC differentiation is a continuous process tightly regulated in the gut by a complex regulatory network. However, the effect of chronic hyperglycemia, in the modulation of regulatory networks controlling identity and differentiation of EECs, has not been analyzed. This study aimed to investigate the effect of glucotoxicity on EEC differentiation in small intestinal organoid platforms. Mouse intestinal organoids were cultured in the presence/absence of high glucose concentrations (35 mM) for 48 h to mimic glucotoxicity. Chronic hyperglycemia impaired the expression of markers related to the differentiation of EEC progenitors (Ngn3) and L-cells (NeuroD1), and it also reduced the expression of Gcg and GLP-1 positive cell number. In addition, the expression of intestinal stem cell markers was reduced in organoids exposed to high glucose concentrations. Our data indicate that glucotoxicity impairs L-cell differentiation, which could be associated with decreased intestinal stem cell proliferative capacity. This study provides the identification of new targets involved in new molecular signaling mechanisms impaired by glucotoxicity that could be a useful tool for the treatment of type 2 diabetes.     

Survey of the interventional cardiology procedures in Italy

Interventional cardiology procedures are increasing because they offer many advantages to patients compared with other techniques: therefore the Italian National Institution for Insurance against Accidents at Work decided to start a survey for monitoring the state-of-the-art regarding the professionals involved in those procedures. The survey covered six cardiology and medical physics Italian departments. Each centre was asked to record 10 examinations for five types of procedures: coronary angiography (CA), electrophysiology studies (ES), pacemaker implantation (PI), percutaneous transluminal coronary angioplasty (PTCA) and radiofrequency catheter ablation (RA). For each examination all the centres were requested to fill in a questionnaire containing information regarding the operator performing the examination, the patient and the procedure. A total of 290 examinations were recorded: 103 CA, 14 ES, 68 PI, 79 PTCA and 26 RA. As occupational doses are strongly related to patient doses, both patients and operators radiation dose data are reported. Ratios of maximum to minimum mean patient doses across the hospitals surveyed were 2.0, 3.9, 7.0, 1.8 and 1.4 for CA, ES, PI, PTCA and RA, respectively. The calculated rounded mean dose-area product values across all participating hospitals were comparable with other values reported in the literature. In general, specific radiation protection tools were used by all operators performing different procedures in all hospitals. A major issue in this survey was the absence of information about correlation between staff and patient doses in a single procedure: future studies could be more aimed to prospective goals where occupational exposures per procedure are monitored specifically.     

Influence of water on bond behavior between CFRP sheet and natural calcareous stones

In this paper the effect of a long term immersion in water on bond durability is analyzed when FRPs (Fiber Reinforced Plastic) are externally applied to a masonry substrate. In the performed research a substrate made by natural calcareous stones, strengthened by CFRP (Carbon Fiber Reinforced Plastic) sheets has been analyzed. For a better comprehension of water effect on the adhesive bond between stone and CFRP, the same treatments were performed to the constituent materials, namely epoxy resins, CFRP sheets and stones. To this aim mechanical tests were carried out on stone, composite materials and epoxy resins before and after their immersion in water, evaluating the effects of this agent on the properties of the materials. The influence of the aging in water on the interface stone-reinforcement was analyzed in terms of bond strength, maximum bond stress, optimal bond length, slip-bond stress relationship and mode of failure. In addition the possibility of calibrating design relationships, taking into account the influence of environmental conditions is discussed. Detailed results on adhesives and composites aged in water have been reported in a previous paper while in the present work the significant decay of the mechanical properties of the stone is specifically investigated. With regard to the conditioning treatment a reduction of the bond strength has been observed (up to 26%) as well as a similar decrease of the maximum bond stress; in addition the aged specimens have shown a more fragile behavior. On the basis of the obtained results the empirical coefficient, reported in the available Italian Guidelines, to determine the FRP-masonry bond strength seems still effective when the system FRP-masonry is aged in water once the properties of the aged materials are considered in the provided relationships.