Immunohistochemical localization and mRNA expression of matrix Gla protein and fetuin-A in bone biopsies of hemodialysis patients

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Identification of maternal uniparental isodisomy of chromosome 10 in a patient with mitochondrial DNA depletion syndrome

Letter to the Edito

Presence of Escherichia coli O157, Salmonella spp., and Listeria monocytogenes in Raw Ovine Milk Destined for Cheese Production and Evaluation of the Equivalence Between the Analytical Methods Applied

Italy is one of the main producers and exporters of cheese made from unpasteurized sheep milk. Since raw milk and its derived products are known sources of human infections, cheese produced from raw sheep milk could pose a microbiological threat to public health. Hence, the objectives of the study were: to characterize the potential risk of the presence of pathogens Escherichia coli O157, Listeria monocytogenes, and Salmonella in raw ovine milk destined for cheese production obtained from all the sheep farms (n = 24) in the Marches region (Central Italy) that directly transform raw milk into cheeses and to evaluate the equivalence between the analytical methods applied. A three-step molecular method (simultaneous culture enrichment, species-specific DNA magnetic isolation, and multiplex real-time polymerase chain reaction) was used for milk (n = 143) and cheese (n = 5) analysis over a 3- year period. L. monocytogenes was not detected on any of the farms, while E. coli O157 was found on three farms, although only using the molecular method. Four farms tested positive for Salmonella spp., and Salmonella enterica subsp. diarizonae serovar 61:k:1,5,7 was isolated in one of those cases. This information highlights the need to develop preventative measures to guarantee a high level of consumer safety for this specific product line, and the molecular method could be a time-saving and sensitive tool to be used in routine diagnosis

Microbiological quality assessment of meals and work surfaces in a school-deferred catering system

Microbiological contamination is one of the main parameters that must be assessed to assure the safety of foods produced by a catering system. The aim of this study conducted between 2011 and 2015 was to assess the microbiological quality of meals (n =620), work surfaces (n =621) and the operators’ hands (n =319) in a school-deferred catering system. A general positive trend regarding the microbial contamination of food samples and surfaces was observed. Salmonella spp. and Listeria monocytogenes were never detected in prepared foods; moreover, the total mesophilic aerobes and the sulfite-reducing clostridia were always below the reference limit. The occurrence of Bacillus cereus in raw and cooked vegetables highlighted the need for corrective action and suggested that continuous monitoring of this toxin-forming microorganism was necessary. A critical evaluation of the data obtained from the microbiological analyses allowed us to obtain an overview of improvements and emerging critical issues

Single-cell sequencing maps gene expression to mutational phylogenies in PDGF- and EGF-driven gliomas

Glioblastoma multiforme (GBM) is the most common and aggressive type of primary brain tumor. Epidermal growth factor (EGF) and platelet‐derived growth factor (PDGF) receptors are frequently amplified and/or possess gain‐of‐function mutations in GBM. However, clinical trials of tyrosine‐kinase inhibitors have shown disappointing efficacy, in part due to intra‐tumor heterogeneity. To assess the effect of clonal heterogeneity on gene expression, we derived an approach to map single‐cell expression profiles to sequentially acquired mutations identified from exome sequencing. Using 288 single cells, we constructed high‐resolution phylogenies of EGF‐driven and PDGF‐driven GBMs, modeling transcriptional kinetics during tumor evolution. Descending the phylogenetic tree of a PDGF‐driven tumor corresponded to a progressive induction of an oligodendrocyte progenitor‐like cell type, expressing pro‐angiogenic factors. In contrast, phylogenetic analysis of an EGFR‐amplified tumor showed an up‐regulation of pro‐invasive genes. An in‐frame deletion in a specific dimerization domain of PDGF receptor correlates with an up‐regulation of growth pathways in a proneural GBM and enhances proliferation when ectopically expressed in glioma cell lines. In‐frame deletions in this domain are frequent in public GBM data

Evaluation of the shelf life and cooking methods of Ascoli-style olives, an Italian specialty food

In recent years, consumers have been increasingly interested in the quality and safety of specialty food products that are perceived as healthy and wholesome. However, few studies have been carried out to assess the effects of domestic cooking on the fate of pathogens in meat-based products. The present study aimed to assess the shelf life of ready-to-cook Ascoli-style olives, an Italian specialty food preparation, through accelerated shelf life tests. A challenge test was also carried out to test the efficacy of cooking methods on the survival of Salmonella Senftemberg strains. Accelerated shelf life assays, including physico-chemical assays (investigating the total volatile basic nitrogen and pH), microbiological counts (Enterobacteriaceae counts) and sensory parameters (appearance, color, odor, flavor and texture) allowed us to obtain sound information on the durability of pre-cooked Ascoli-style olives in an affordable and efficient manner. This is therefore an effective method that could be exploited by food business operators hoping to provide scientific information on product shelf life assessments. The challenge tests demonstrated that fan-baking in an electric oven at 200 °C for 12 min. and deep frying for 4 min. are the more effective treatments needed to inactivate the artificially inoculated Salmonella Senftemberg strains

Single-cell profiling of human gliomas reveals macrophage ontogeny as a basis for regional differences in macrophage activation in the tumor microenvironment

Abstract Background Tumor-associated macrophages (TAMs) are abundant in gliomas and immunosuppressive TAMs are a barrier to emerging immunotherapies. It is unknown to what extent macrophages derived from peripheral blood adopt the phenotype of brain-resident microglia in pre-treatment gliomas. The relative proportions of blood-derived macrophages and microglia have been poorly quantified in clinical samples due to a paucity of markers that distinguish these cell types in malignant tissue. Results We perform single-cell RNA-sequencing of human gliomas and identify phenotypic differences in TAMs of distinct lineages. We isolate TAMs from patient biopsies and compare them with macrophages from non-malignant human tissue, glioma atlases, and murine glioma models. We present a novel signature that distinguishes TAMs by ontogeny in human gliomas. Blood-derived TAMs upregulate immunosuppressive cytokines and show an altered metabolism compared to microglial TAMs. They are also enriched in perivascular and necrotic regions. The gene signature of blood-derived TAMs, but not microglial TAMs, correlates with significantly inferior survival in low-grade glioma. Surprisingly, TAMs frequently co-express canonical pro-inflammatory (M1) and alternatively activated (M2) genes in individual cells. Conclusions We conclude that blood-derived TAMs significantly infiltrate pre-treatment gliomas, to a degree that varies by glioma subtype and tumor compartment. Blood-derived TAMs do not universally conform to the phenotype of microglia, but preferentially express immunosuppressive cytokines and show an altered metabolism. Our results argue against status quo therapeutic strategies that target TAMs indiscriminately and in favor of strategies that specifically target immunosuppressive blood-derived TAMs