MicroRNAs-Proteomic Networks Characterizing Human Medulloblastoma-SLCs

Medulloblastoma (MB) is the most common malignant brain tumor of pediatric age and is characterized by cells expressing stem, astroglial, and neuronal markers. Among them, stem-like cells (hMB-SLCs) represent a fraction of the tumor cell population with the potential of self-renewal and proliferation and have been associated with tumor poor prognosis. In this context, microRNAs have been described as playing a pivotal role in stem cells differentiation. In our paper, we analyze microRNAs profile and genes expression of hMB-SLCs before and after Retinoic Acid- (RA-) induced differentiation. We aimed to identify pivotal players of specific pathways sustaining stemness and/or tumor development and progression and integrate the results of our recent proteomic study. Our results uncovered 22 differentially expressed microRNAs that were used as input together with deregulated genes and proteins in the Genomatix Pathway System (GePS) analysis revealing 3 subnetworks that could be interestingly involved in the maintenance of hMB-SLCs proliferation. Taken together, our findings highlight microRNAs, genes, and proteins that are significantly modulated in hMB-SLCs with respect to their RA-differentiated counterparts and could open new perspectives for prognostic and therapeutic intervention on MB

a metaproteomic pipeline to identify newborn mouse gut phylotypes

Abstract In order to characterize newborn mouse gut microbiota phylotypes in very early-life stages, an original metaproteomic pipeline, based on LC–MS 2 -spectra and Mascot driven NCBI non-redundant repository database interrogation was developed. An original computational analysis assisted in the generation of a taxonomic gut architecture from protein hits to operational taxonomic units (OTUs) and related functional categories. Regardless of the mouse's genetic background, a prevalence of Firmicutes (Lactobacillaceae) and Proteobacteria (Enterobacteriaceae) was observed among the entire Eubacteria taxonomic node. However, a higher abundance of Firmicutes was retrieved for Balb/c gut microbiota compared to Rag2 ko mice, the latter was mainly characterized by a Proteobacteria enriched microbiota. The metaproteomic-obtained OTUs were supported, for the identification (ID) of the cultivable bacteria fraction, corroborated by axenic culture-based MALDI-TOF MS IDs. Particularly, functional analysis of Rag2 ko mice gut microbiota proteins revealed the presence of abundant glutathione, riboflavin metabolism and pentose phosphate pathway components, possibly related to genetic background. The metaproteomic pipeline herein presented may represent a useful tool to investigate the highly debated onset of the human gut microbiota in the first days of life, when the bacterial composition, despite its very low diversity (complexity), is still very far from an exhaustive description and other complex microbial consortia. Biological significance The manuscript deals with a "frontier" topic regarding the study of the gut microbiota and the application of a metaproteomic pipeline to unveil the complexity of this fascinating ecosystem at the very early stages of life. Indeed during these phases, its diversity is very low but the bacterial content is highly "instable", and the relative balance between mucosal and fecal bacteria starts its dynamics of "fight" to get homeostasis. However, in the neonatal period, especially immediately after birth, a comprehensive description of this microbial eco-organ is still lacking, while it should be mandatory to highlight its first mechanisms of homeostasis and perturbation, while it co-develops with and within the host species. In order to unravel its low but almost unknown microbial community multiplicity, the newborn mouse gut, characterized by a "very" low complexity, was herein selected as model to design a LC–MS 2 -based shotgun metaproteomic approach, potentially suitable to study onset and shaping in human newborns. A microbiological semi-automatic computational analysis was performed to infer gut phylotypes; such as proof of evidence, related OTUs were compared to axenic-culture-based MALDI-TOF MS IDs showing consistency at family and phyla levels for the bacterial cultivable fraction. This article is part of a Special Issue entitled: Trends in Microbial Proteomics

Influencia de la dieta con taninos y del almacenamiento refrigerado sobre la calidad sensorial del músculo semitendinosus cocido sous – vide de vaca de refugo

En Argentina, la categoría vaca de refugo se considera un subproducto y se comercializa a bajo valor. Sin embargo, el 50% de los kg vendidos de la producción nacional actual corresponden a la categoría vacuna de hembras. Por un lado, esta situación lleva al sector ganadero a buscar una oportunidad para generar alternativas en la obtención de productos que permitan aumentar su valor. Los taninos son compuestos polifenólicos de origen vegetal usados en la producción bovina como forraje o como suplemento. Estos, en dietas suministradas a rumiantes, permiten mejorar la ganancia de peso diario, son aditivos naturales, no presentan restricciones en su utilización como otros, y podrían impactar positivamente en la calidad de la carne. Por otro lado, en respuesta a la demanda del consumidor de alimentos listos para consumir, uno de los procesos más usados por la industria es el sous-vide (SV), que consiste en “cocción al vacío”, seguido de almacenamiento refrigerado, permitiendo dar un importante agregado de valor al producto. El objetivo del presente trabajo fue evaluar el efecto suplementación con taninos en alimentación de vacas de refugo y el tiempo de almacenamiento refrigerado sobre los atributos sensoriales del músculo semitendinosus (SD) procesado mediante cocción SV. Se realizaron dos tratamientos, suplementación con taninos (SCT): ensilaje de sorgo con suplementación energética / proteica con agregado de taninos de quebracho colorado (Schinopsis balansae) a una ración de 2 g/100 g materia seca y suplementación sin taninos (SST): ensilaje de sorgo con suplementación energética / proteica sin agregado de taninos. En cada tratamiento se utilizaron 12 vacas de refugo Hereford (6 mm EGD) faenadas en un frigorífico local. De cada media res izquierda, se tomaron los músculos SD los cuales se envasaron y congelaron. Para el proceso SV, los músculos se descongelaron (1 ± 0,5 °C), se acondicionaron y se envasaron al vacío. La cocción se realizó en un equipo autoclave hasta alcanzar 70 °C - 2 min en el punto de mínimo tratamiento. Posteriormente, se realizó el almacenamiento refrigerado (1 ± 0,5 °C) durante 0, 7, 14 y 21 días. El perfil sensorial del producto cocido se realizó mediante un análisis descriptivo cuantitativo. Los resultados se analizaron estadísticamente mediante análisis de varianza de dos factores y prueba de comparación de medias de Tukey (P0,05) en el flavor, aroma, tejido conectivo y jugosidad. La interacción entre la suplementación y el tiempo de almacenamiento refrigerado resultó significativa para los atributos de terneza inicial y sostenida. No obstante, no se observaron diferencias en estos atributos, para SST y SCT, entre el inicio y el final del tiempo de almacenamiento refrigerado. En general, durante todo el tiempo de almacenamiento refrigerado, los valores de flavor y aroma del producto cocido se mantuvieron en el rango 4 a 5.In Argentina, the cull cow category is considered a by-product and It is sold at a low Price. However, 50% of the kilograms sold of the current national production of beef correspond to the female category. This situation carries the livestock sector to look for an opportunity to generate alternatives to obtain obtaining products that increase the value of this bovine category. Tannins are polyphenolic compounds of vegetable origins used as forage or as a supplement in bovine production. They allow to improve daily weight increase, they are natural additives and they do not have restrictions when they are used like others have. In the diets supplied to ruminants tannins They could have a positive impact on the quality of the meat. On the other hand, in response to consumer demand for ready-to-eat food, one of the options used by industry is the sous vide process (SV), which consists in “vacuum cooking” followed by refrigerated storage which allows to add a huge value to the product. The aim of this study was to evaluate the effect of tannin supplementation in the cull cow diet and the length of refrigerated storage on the sensory attributes in the semitendinosus muscle (SD) processed through SV cooking technology. Two treatments were carried out, supplementation with tannins (SCT): sorghum silage with energy / protein supplementation of quebracho (Schinopsis balansae) tannins 2 g/100 g dry matter and supplementation without tannins (SST): sorghum silage with energy / protein supplementation without tannins. In each treatment, 12 Hereford cull cows were used and after they reached the desired body condition (6 mm EGD) they were slaughtered in a local slaughterhouse. SD muscles from left side were removed and frozen. For SV processing, SD muscles were thawed (1 ± 0.5°C), conditioned and vacuum packed in bags suitable for cooking. Cooking was carried out in an autoclave until 70°C - 2 min were reached at the product minimum treatment point (PMT). Subsequently, refrigerated storage (1 ± 0.5°C) was carried out for 0, 7, 14 and 21 days. The sensory profile was made through a quantitative descriptive analysis and the results were analyzed statistically for two-factors variance and Tukey’s mean comparison test (level of significance of 0,05). Flavor, aroma, connective tissue and juiciness were not affected (P>0,05). The interaction between supplementation and refrigerated storage time was significant for initial and sustained tenderness attributes. However, no differences were observed between initial and final refrigerated storage time on initial and sustained tenderness. In general, during all the refrigerated storage time, the cooked beef flavor and aroma scores were kept between 4 and 5.EEA ConcordiaFil: Urbani, Valeria Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concordia. Agencia De Extensión Rural Chajarí; ArgentinaFil: Biolatto, Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay; Argentina.Fil: Palladino, Pablo Martin. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto Tecnología de Alimentos; Argentin

Food Safety: Secretome of Lactococcus lactis and Listeria monocytogenes in competition.

Listeria monocytogenes (LM) is a foodborne pathogen responsible of listeriosis. In the spreading of this pathology, milk and dairy products are key reservoir for this pathogen1. Food processing represents one of the major steps that could be linked to LM growth. Inhibition of LM growth through competition of Lactococcus lactis (LAC) could represent a solution to this problem. Exoproteome of LM and two different strains of Lactic Acid Bacteria in co-culture have been studied in order to highlight mechanisms of bacterial competition useful to improve food safety. Two different strains of LAC and one strain of LM were cultivated in appropriate medium cultures (BHI), also in competition. Filtrated cultures (SECRETOME) were lyophilized and resuspended for proteomics analysis. Shotgun analysis on each secretome was performed on nano UPLC-MS system. Obtained data reveal, during competition, the higher production by LM of moonlighting protein Enolase and Glucose 6 Phosphate isomerase, of Septation ring formation regulator EzrA, involved into cell replication and the lower secretion of Endopeptidase P60. In parallel, L. lactis produced higher amounts of Secreted 45 kDa protein and switched from lantibiotic Nisin A production to Nisin Z production. In competition with LM, LAC strain investigated produce higher amounts of Secreted 45 kDa protein with peptidoglycan lytic activity and the selective secretion of Nisin Z probably to improve lantibiotic solubility in less acidic environment. Next step will be validation of obtained results in dairy products. These results are of interesting to design new strategies of fighting LM as contaminant in food from animal origin.Work supported by Ministry of Health-CCM “Milano EXPO 2015 Project: Garantire la sicurezza alimentare- Valorizzare le produzioni

The Role of Obesity, Body Composition, and Nutrition in COVID-19 Pandemia: A Narrative Review

The coronavirus disease 2019 (COVID-19) pandemic has spread worldwide, infecting nearly 500 million people, with more than 6 million deaths recorded globally. Obesity leads people to be more vulnerable, developing worse outcomes that can require hospitalization in intensive care units (ICU). This review focused on the available findings that investigated the link between COVID-19, body composition, and nutritional status. Most studies showed that not only body fat quantity but also its distribution seems to play a crucial role in COVID-19 severity. Compared to the body mass index (BMI), visceral adipose tissue and intrathoracic fat are better predictors of COVID-19 severity and indicate the need for hospitalization in ICU and invasive mechanical ventilation. High volumes of epicardial adipose tissue and its thickness can cause an infection located in the myocardial tissue, thereby enhancing severe COVID-related myocardial damage with impairments in coronary flow reserve and thromboembolism. Other important components such as sarcopenia and intermuscular fat augment the vulnerability in contracting COVID-19 and increase mortality, inflammation, and muscle damage. Malnutrition is prevalent in this population, but a lack of knowledge remains regarding the beneficial effects aimed at optimizing nutritional status to limit catabolism and preserve muscle mass. Finally, with the increase in patients recovering from COVID-19, evaluation and treatment in those with Long COVID syndrome may become highly relevant

Arterial Stiffness, Subendocardial Impairment, and 30-Day Readmission in Heart Failure Older Patients

Arterial stiffness and subendocardial perfusion impairment may play a significant role in heart failure (HF) outcomes. The aim of the study was to examine the main predictors of 30-day readmission in geriatric patients, hospitalized with HF, explore hemodynamical parameters, arterial stiffness indexes, and subendocardial viability ratio (SEVR). In total, 41 hospitalized patients, affected by HF, were included; they underwent clinical evaluation, routine laboratory testing, and echocardiography. At the time of admission, after the achievement of clinical stability (defined as switching from intravenous to oral diuretic therapy), and at discharge, arterial tonometry was performed to evaluate carotid-femoral pulse wave velocity (PWVcf) and SEVR (then corrected for hemoglobin concentration and oxygen saturation). Through the evaluations, a significant progressive decrease in PWVcf was described (17.79 ± 4.49, 13.54 ± 4.54, and 9.94 ± 3.73 m/s), even after adjustment for age, gender, mean arterial pressure (MAP) variation, and left ventricular ejection fraction (LVEF). A significant improvement was registered for both SEVR (83.48 ± 24.43, 97.94 ± 26.84, and 113.29 ± 38.02) and corrected SEVR (12.74 ± 4.69, 15.71 ± 5.30, and 18.55 ± 6.66) values, and it was still significant when adjusted for age, gender, MAP variation, and LVEF. After discharge, 26.8% of patients were readmitted within 30 days. In a multivariate binary logistic regression analysis, PWVcf at discharge was the only predictor of 30-day readmission (odds ratio [OR] 1.957, 95% CI 1.112-3.443). In conclusion, medical therapy seems to improve arterial stiffness and subendocardial perfusion in geriatric patients hospitalized with heart failure. Furthermore, PWVcf is a valid predictor of 30-day readmission. Its feasibility in clinical practice may provide an instrument to detect patients with HF at high risk of rehospitalization

Identification of two novel LDLR variants by Next Generation Sequencing

Introduction. Familial hypercholesterolemia (FH) is an autosomal dominant inherited disease characterized by elevated plasma low-density lipoprotein cholesterol (LDL-C). Targeted Next Generation Sequencing (NGS) is a new opportunity to expand the existing pathogenic variants (PVs) spectrum associated to FH. Our aim was to report a diagnostic NGS-based approach to detect variants associated to FH.Methods. We report two patients: a 48-year-old Asian woman, without known history of hypercholesterolemia and a 46-year-old Caucasian man, with childhood hypercholesterolemia.Results. An effective NGS-based pipeline, FH-Devyser kit/Amplicon Suite, beginning from sequencing to data analysis, did not identify known PVs in the LDLR, APOB, APOE, LDLRAP1, STAP1 and PCSK9 genes, but revealed two novel LDLR variants (c.1564A>T, p.Ile522Phe and c.1688C>T, p.Pro563Leu).Discussion and conclusions. This study showed that an effective NGS-based pipeline led to a definitive diagnosis in two FH families, allowing to plan their therapeutic treatment. Although the functional consequence of the two LDLR variants needs to be assessed in vitro, the in silico analysis and high preservation of the two amino acid positions observed in the LDLR protein, across different animal species, suggest that both variants are deleterious

Glutathione transferases and glutathionylated hemoglobin in workers exposed to low doses of 1,3-butadiene.

We evaluated glutathione transferase (GST) activities and the levels of glutathionylated hemoglobin in the RBCof 42 workers exposed to 1,3-butadiene in a petrochemical plant, using 43 workers not exposed to 1,3-butadiene and 82 foresters as internal and external controls, respectively. Median 1,3-butadiene exposure levels were 1.5, 0.4, and 0.1 Mg/m3 in 1,3-butadieneexposed workers, in workers not directly exposed to 1,3-butadiene, and in foresters, respectively. In addition, we determined in the peripheral blood lymphocytes of the same individuals the presence of GST polymorphic genes GSTT1 and GSTM1 and the distribution of GSTP1 allelic variants. Comparing the mean values observed in petrochemical workers with those of control foresters, we found a marked decrease of GST enzymatic activity and a significant increase of glutathionylated hemoglobin in the petrochemical workers. A weak but significant negative correlation was found between levels of 1,3-butadiene exposure and GST activity, whereas a positive correlation was found between 1,3-butadiene exposure and glutathionylated hemoglobin. A negative correlation was also observed between GST activity and glutathionylated hemoglobin. No influence of confounders was observed. Using a multiple linear regression model, up to 50.6% and 41.9% of the variability observed in glutathionylated hemoglobin and GST activity, respectively, were explained by 1,3- butadiene exposure, working setting, and GSTT1 genotype. These results indicate that occupational exposure to 1,3-butadiene induces an oxidative stress that impairs the GST balance in RBC, and suggest that GST activity and glutathionylated hemoglobin could be recommended as promising biomarkers of effect in petrochemical workers