Tumor Necrosis Factor Receptor SF10A (TNFRSF10A) SNPs Correlate With Corticosteroid Response in Duchenne Muscular Dystrophy

Background Duchenne muscular dystrophy (DMD) is a rare and severe X-linked muscular dystrophy in which the standard of care with variable outcome, also due to different drug response, is chronic off-label treatment with corticosteroids (CS). In order to search for SNP biomarkers for corticosteroid responsiveness, we genotyped variants across 205 DMD-related genes in patients with differential response to steroid treatment. Methods and Findings We enrolled a total of 228 DMD patients with identified dystrophin mutations, 78 of these patients have been under corticosteroid treatment for at least 5 years. DMD patients were defined as high responders (HR) if they had maintained the ability to walk after 15 years of age and low responders (LR) for those who had lost ambulation before the age of 10 despite corticosteroid therapy. Based on interactome mapping, we prioritized 205 genes and sequenced them in 21 DMD patients (discovery cohort or DiC = 21). We identified 43 SNPs that discriminate between HR and LR. Discriminant Analysis of Principal Components (DAPC) prioritized 2 response-associated SNPs in theTNFRSF10Agene. Validation of this genotype was done in two additional larger cohorts composed of 46 DMD patients on corticosteroid therapy (validation cohorts or VaC1), and 150 non ambulant DMD patients and never treated with corticosteroids (VaC2). SNP analysis in all validation cohorts (N= 207) showed that the CT haplotype is significantly associated with HR DMDs confirming the discovery results. Conclusion We have shown that TNFRSF10A CT haplotype correlates with corticosteroid response in DMD patients and propose it as an exploratory CS response biomarker

Viscoelastic analysis of sugarcane stems through mechanical tests and moiré techniques

Orientadores: Inácio Maria Dal Fabbro, Jonathan GazzolaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia AgrícolaResumo: A determinação das propriedades mecânicas de materiais biológicos, em especial os espécimes vegetais, é complexa em função de sua anisotropia, presença de umidade, composição celular interna não homogênea, não linearidade geométrica, entre outras características que não atendem aos pressupostos básicos da Mecânica dos Meios Contínuos. Espécimes de cana-de-açúcar em virtude do alto teor de fibras e líquidos em seu tecido vegetal são modelados, no que concerne ao comportamento mecânico pela Teoria da Viscoelasticidade. Por meio da realização de ensaios mecânicos de compressão uniaxial e restritos, amostras cilíndricas, da porção internodal do parênquima de cana-de-açúcar foram submetidas a carregamentos em prensa universal a diferentes taxas de deformação (frequência) e os resultados tensão-deformação contribuem para a obtenção das funções viscoelásticas dependentes do tempo, notadamente, E(t),?(t), ?(t) e ?(t). O desenvolvimento das tensões e deformações de contato e sua distribuição em corpos submetidos a carregamento diametral (ensaio Lobo-Carneiro) é verificado comparativamente com outros materiais (madeira e borracha) pela técnica interferométrica de moiré de sombraAbstract: The determination of the mechanical properties, as well as of the mechanical behavior of biological materials, particularly of vegetative materials is quite complex due its composition which includes fibers and other solids as well as liquid and gas phases. That composition generates material nonlinearities which associated with geometrical nonlinearities do not meet the basic requirements of the Mechanics of Continuum Media. Specimens of sugarcane, due its high fiber, liquid and gases content are better modeled, concerning mechanical behavior, by the Theory of Linear Viscoelasticity. In this research work the uniaxial stress and strain as well as constrained tests carried on the sugarcane internode parenchyma allowed the generation of the viscoelastics functions named E(t),?(t), ?(t) e ?(t). These were later on expressed as Creep and Relaxation Functions. Contact stress and strain occurrence on on cylindrical specimens were studied through the Lobo Carneiro (Brazilian Test) tests. Moiré optical techniques were employed to generate strain and strain distribution images in the axial and diametrical compressions on sugar cane internode, wood specimens as well as on ruber cylinders as a matter of illustration and comparisonDoutoradoMáquinas AgrícolasDoutor em Engenharia AgrícolaCNP

A Role of Tumor-Released Exosomes in Paracrine Dissemination and Metastasis

Metastatic diffusion is thought to be a multi-step phenomenon involving the release of cells from the primary tumor and their diffusion through the body. Currently, several hypotheses have been put forward in order to explain the origin of cancer metastasis, including epithelial⁻mesenchymal transition, mutagenesis of stem cells, and a facilitating role of macrophages, involving, for example, transformation or fusion hybridization with neoplastic cells. In this paradigm, tumor-secreted extracellular vesicles (EVs), such as exosomes, play a pivotal role in cell communications, delivering a plethora of biomolecules including proteins, lipids, and nucleic acids. For their natural role in shuttling molecules, EVs have been newly considered a part of the metastatic cascade. They have a prominent role in preparing the so-called “tumor niches„ in target organs. However, recent evidence has pointed out an even more interesting role of tumor EVs, consisting in their ability to induce malignant transformation in resident mesenchymal stem cells. All in all, in this review, we discuss the multiple involvements of EVs in the metastatic cascade, and how we can exploit and manipulate EVs in order to reduce the metastatic spread of malignant tumors

Prevalence, Molecular Characterization and Antimicrobial Susceptibility of Clostridioides difficile Isolated from Pig Carcasses and Pork Products in Central Italy

In the last decade, the incidence and severity of Clostridioides difficile infections (CDIs) in humans have been increasing and community-associated infections have been described. For these reasons, the interest in C. difficile in food and in food animals has increased, suggesting other possible sources of C. difficile acquisition. This study evaluated the presence of C. difficile on pig carcasses at the slaughterhouse and in pork products in Central Italy. The contamination rate on pig carcasses was 4/179 (2.3%). Regarding food samples, a total of 216 pork products were tested (74 raw meat preparations and 142 ready-to-eat food samples made by cured raw meat). The real-time PCR screening was positive for 1/74 raw meat preparation (1.35%) and for 1/142 ready-to-eat food samples (0.7%) C. difficile was isolated only from the raw meat preparation (pork sausage). All the isolated strains were toxigenic and susceptible to all the tested antibiotics. Strains isolated from carcass samples displayed A+B+CDTa+CDTb+ profile, were toxinotype IV and belonged to the same ribotype arbitrary named TV93, while the one isolated from food samples displayed A+B+CDTa-CDTb- profile and it was not possible to determine ribotype and toxinotype, because it was lost after freeze storage. It was concluded that the prevalence of C. difficile in the pork supply chain is very low

Hazelnut Shells as Source of Active Ingredients: Extracts Preparation and Characterization

Hazelnut shells represent a waste material (about 42% of the total biomass) deriving from hazelnut harvest. These are mainly used as a heating source; however, they represent an interesting source of polyphenols useful in health field. The impact on phenolic profile and concentrations of hazelnut shell extracts obtained by three extraction methods (maceration, ultrasonic bath, and high-power ultrasonic), as well as temperature, extraction time, and preventive maceration, was studied. The prepared extracts were characterized in terms of chemical composition, antioxidant and antimicrobial activities. Eighteen different phenolic compounds were identified and quantified by chemical analysis and gallic acid was the most abundant in all the extracts analyzed. Other relevant compounds were chlorogenic acid, protocatechuic acid and catechin. Preventive maceration had a positive effect on the extraction of different types of compounds regardless of the method performed. Application of the high-power ultrasonic method had different effects, either positive or negative, depending on the type of compound and extraction time. All the prepared extracts showed antioxidant activity especially those prepared by maceration, and many of them were able to inhibit the growth of both B. subtilis and B. cereus.15n

Acridine Orange/exosomes increase the delivery and the effectiveness of Acridine Orange in human melanoma cells: A new prototype for theranostics of tumors

Specifically targeted drug delivery systems with low immunogenicity and toxicity are deemed to increase efficacy of cancer chemotherapy. Acridine Orange (AO) is an acidophilic dye with a strong tumoricidal action following excitation with a light source at 466 nm. However, to date the clinical use of AO is limited by the potential side effects elicited by systemic administration. The endogenous nanocarrier exosomes have been recently introduced as a natural delivery system for therapeutic molecules. In this article, we show the outcome of the administration to human melanoma cells of AO charged Exosomes (Exo-AO), in both monolayer and spheroid models. The results showed an extended drug delivery time of Exo-AO to melanoma cells as compared to the free AO, improving the cytotoxicity of AO. This study shows that Exo-AO have a great potential for a real exploitation as a new theranostic approach against tumors based on AO delivered through the exosomes

Bdellovibrio bacteriovorus to control Escherichia coli on meat matrices

Bdellovibrio bacteriovorus is a predator micro-organism towards other Gram-negative bacteria. We tested B. bacteriovorus to control Escherichia coli growth on chicken slices and canned beef. Moreover, we analysed B. bacteriovorus’s lytic ability on eight toxigenic or multidrug-resistant E. coli strains. In chicken slices, the predator induced the highest prey reduction (4.3 log) respect to control at 6 h. In canned beef, the predator induced the highest prey reduction (2.1 log) respect to control at 6 h. Moreover, B. bacteriovorus showed lytic ability towards all tested E. coli strains. B. bacteriovorus could control E. coli and other pathogenic and spoilage bacteria in those meat-based foods that have a shelf life <10 days. It could integrate modified atmosphere packaging (MAP) to prolong the shelf life and improve the safety of prepacked fresh meat, meat preparations and meat products. In future applications on meat-based foods, B. bacteriovorus could also minimise the use of additives

POPDC1(S201F) causes muscular dystrophy and arrhythmia by affecting protein trafficking.

The Popeye domain-containing 1 (POPDC1) gene encodes a plasma membrane-localized cAMP-binding protein that is abundantly expressed in striated muscle. In animal models, POPDC1 is an essential regulator of structure and function of cardiac and skeletal muscle; however, POPDC1 mutations have not been associated with human cardiac and muscular diseases. Here, we have described a homozygous missense variant (c.602C>T, p.S201F) in POPDC1, identified by whole-exome sequencing, in a family of 4 with cardiac arrhythmia and limb-girdle muscular dystrophy (LGMD). This allele was absent in known databases and segregated with the pathological phenotype in this family. We did not find the allele in a further screen of 104 patients with a similar phenotype, suggesting this mutation to be family specific. Compared with WT protein, POPDC1(S201F) displayed a 50% reduction in cAMP affinity, and in skeletal muscle from patients, both POPDC1(S201F) and WT POPDC2 displayed impaired membrane trafficking. Forced expression of POPDC1(S201F) in a murine cardiac muscle cell line (HL-1) increased hyperpolarization and upstroke velocity of the action potential. In zebrafish, expression of the homologous mutation (popdc1(S191F)) caused heart and skeletal muscle phenotypes that resembled those observed in patients. Our study therefore identifies POPDC1 as a disease gene causing a very rare autosomal recessive cardiac arrhythmia and LGMD, expanding the genetic causes of this heterogeneous group of inherited rare diseases.peerReviewe