Epigenetic Mechanisms Histone Deacetylase-Dependent Regulate the Glioblastoma Angiogenic Matrisome and Disrupt Endothelial Cell Behavior In Vitro.

Glioblastoma (GBM) is the most aggressive brain tumor and different efforts have been employed in the search for new drugs and therapeutic protocols for GBM. Epitranscriptomics has shed light on new druggable Epigenetic therapies specifically designed to modulate GBM biology and behavior such as Histone Deacetylase inhibitors (iHDAC). Although the effects of iHDAC on GBM have been largely explored, there is a lack of information on the underlaying mechanisms HDAC-dependent that modulate the repertoire of GBM secreted molecules focusing on the set of Extracellular Matrix (ECM) associated proteins, the Matrisome, that may impact the surrounding tumor microenvironment. To acquire a better comprehension of the impacts of HDAC activity on the GBM Matrisome, we studied the alterations on the Matrisome-associated ECM regulators, Core Matrisome ECM glycoproteins, ECM-affiliated proteins and Proteoglycans upon HDAC inhibition in vitro as well as their relationship with glioma pathophysiological/clinical features and angiogenesis. For this, U87MG GBM cells were treated for with iHDAC or vehicle (control) and the whole secretome was processed by Mass Spectrometry NANOLC-MS/MS. In silico analyses revealed that proteins associated to the Angiogenic Matrisome (AngioMatrix), including Decorin, ADAM10, ADAM12 and ADAM15 were differentially regulated in iHDAC versus control secretome. Interestingly, genes coding for the Matrisome proteins differentially regulated were found mutated in patients and were correlated to glioma pathophysiological/clinical features. In vitro functional assays, using HBMEC endothelial cells exposed to the secretome of control or iHDAC treated GBM cells, coupled to 2D and 3D GBM cell culture system, showed impaired migratory capacity of endothelial cells and disrupted tubulogenesis in a Fibronectin and VEGF independent fashion. Collectively, our study provides understanding of epigenetic mechanisms HDAC-dependent to key Matrisomal proteins that may contribute to identify new druggable Epigenetic therapies or gliomagenesis biomarkers with relevant implications to improve therapeutic protocols for this malignancy.

Validation of a Satisfaction Scale with a Telemedicine COVID-19 Service: Satis-COVID.

Objectives: Despite being a widespread tool, telehealth was significantly incorporated during the COVID-19 pandemic period, but it still lacks analysis methodologies, greater digital security, and satisfaction assessment instruments that are still little explored and validated. The objective is to assess user satisfaction through the validation of a satisfaction scale with a telemedicine COVID-19 service (TeleCOVID). Methods: Cross-sectional study of a cohort of confirmed COVID-19 cases evaluated and monitored by the TeleCOVID team. To study the scale's measurement qualities, a factorial analysis was performed to test the validity of the construct. Correlation between items and the global scale was assessed using Spearman's correlation coefficient, and the instrument's internal consistency was assessed using Cronbach's alpha coefficient. Results: There were 1,181 respondents evaluating the care received from the TeleCOVID project. A total of 61.6% were female, and 62.4% aged between 30 and 59 years. The correlation coefficients indicated a good correlation between the items present in the instrument. The internal consistency of the global scale was high (Cronbach's alpha = 0.903) and the item-total correlations for the scale ranged from 0.563 to 0.820. The average overall user satisfaction was 4.58, based upon a 5-point Likert scale where 5 is the highest level of satisfaction. Conclusions: The results presented here show how much telehealth can contribute to improving access, resolutibility, and quality of care to the population in general in Public Health Care. In view of the results found, it can be said that the TeleCOVID team offered excellent care and fulfilled its proposed objectives. The scale fulfills its objective of evaluating the quality of teleservice, bringing good results in terms of validity and reliability, in addition to showing high levels of user satisfaction.

O-GlcNAcylation protein disruption by Thiamet G promotes changes on the GBM U87-MG cells secretome molecular signature.

Glioblastoma (GBM) is a grade IV glioma highly aggressive and refractory to the therapeutic approaches currently in use. O-GlcNAcylation plays a key role for tumor aggressiveness and progression in different types of cancer; however, experimental evidence of its involvement in GBM are still lacking. Here, we show that O-GlcNAcylation plays a critical role in maintaining the composition of the GBM secretome, whereas inhibition of OGA activity disrupts the intercellular signaling via microvesicles. Using a label-free quantitative proteomics methodology, we identified 51 proteins in the GBM secretome whose abundance was significantly altered by activity inhibition of O-GlcNAcase (iOGA). Among these proteins, we observed that proteins related to proteasome activity and to regulation of immune response in the tumor microenvironment were consistently downregulated in GBM cells upon iOGA. While the proteins IGFBP3, IL-6 and HSPA5 were downregulated in GBM iOGA cells, the protein SQSTM1/p62 was exclusively found in GBM cells under iOGA. These findings were in line with literature evidence on the role of p62/IL-6 signaling axis in suppressing tumor aggressiveness and our experimental evidence showing a decrease in radioresistance potential of these cells. Taken together, our findings provide evidence that OGA activity may regulate the p62 and IL-6 abundance in the GBM secretome. We propose that the assessment of tumor status from the main proteins present in its secretome may contribute to the advancement of diagnostic, prognostic and even therapeutic tools to approach this relevant malignancy.

Yeasts isolated from Brazilian fermented foods in the protection against infection by pathogenic food bacteria.

The consumption of probiotics has increased due to the reported health benefits, mainly in preventing or treating gastrointestinal pathology. This study investigated the antimicrobial capacity of yeasts, Saccharomyces cerevisiae and Pichia kluyveri, previously isolated from fermented foods (indigenous beverage, kefir and cocoa) against the adhesion of foodborne pathogens to Caco-2 cells. Co-aggregation of yeasts with pathogens and were evaluated by quantitative analysis and using scanning electron and laser confocal microscopies. All yeasts strains were able to co-aggregate with the tested pathogens, however, this activity was strain-dependent. The inhibition tests showed that the adhesion of Escherichia coli EPEC, Listeria monocytogenes and Salmonella Enteritidis to Caco-2 was reduced by all the yeasts studied. Most of the evaluated yeasts showed inhibition rates equal to or greater than the commercial probiotic Saccharomyces boulardii. The yeasts were able to reduce up to 50% of the bacterial infection, as observed for CCMA0615 towards EPEC in exclusion assay; CCMA0731, CCMA0732 and CCMA0615 towards L. monocytogenes in exclusion and competition assays; and CCMA0731 in exclusion and CCMA0731, CCMA0732, CCMA0615 in competition assay towards S. Enteritidis. No antimicrobial compounds were produced by the yeasts, showing that competition for nutrients and/or receptors in the intestinal mucosa was the mechanism to bacterial inhibition.

Human bocavirus in acute gastroenteritis in children in Brazil.

Epidemiological surveillance for Human Bocavirus (HBoV) was conducted on 105 fecal specimens from children with acute gastroenteritis in Bahia, Brazil. Among of a total 105 stool samples, 44 samples were positive for HBoV as detected by nested-PCR. Of the 44 positive samples, co-infections with other enteric viruses (Norovirus, Adenovirus, and Rotavirus) were found in 12 pediatric patients. Mixed infections among HBoV with Norovirus were frequently observed in this population. The phylogenetic analysis identified the presence of HBoV-1, and HBoV 2A species. This study shows that HBoV is another viral pathogen in the etiology of acute gastroenteritis in children in Bahia, Brazil.

Psychometric properties of the Latino Students Patient Safety Questionnaire, Brazilian version.

OBJECTIVES: to assess the psychometric properties of the Latino Students Patient Safety Questionnaire, Brazilian version. METHODS: a methodological study, carried out between April 2020 and January 2021, with 218 nursing and medicine students. Structural and discriminant construct validity were assessed by confirmatory factor analysis and cross factor loadings. Reliability was verified by McDonald's omega, average variance extracted, composite reliability, and item-total correlation. RESULTS: the final model reproduced the original structure of 21 items, distributed in five dimensions, requiring the exclusion of one item. Acceptable fit indexes were obtained (x2/gl=2.325; CFI=0.99; TLI=0.98; RMSEA=0.054). Discriminant validity was confirmed. Reliability indicators were adequate, except McDonald's omega, in one factor (0.68), and average variance extracted, in two factors (0.41; 0.47). CONCLUSIONS: the instrument demonstrated evidence of internal validity and satisfactory reliability among nursing and medical students.

Sensory and flavor-aroma profiles of passion fruit juice fermented by potentially probiotic Lactiplantibacillus plantarum CCMA 0743 strain.

Several non-dairy probiotic beverages are already available to consumers and have been considered suitable carriers for probiotic bacteria. This study aimed to investigate the effect of Lactiplantibacillus plantarum CCMA 0743 in single and co-culture on the volatile compounds and sensory profiles of fermented passion fruit juice. The viability of strains inoculated in juice and MRS matrices was evaluated in a simulated gastrointestinal condition. The bacterial viability after 28 days of refrigerated storage of the juices was also evaluated. L. plantarum CCMA 0743 showed high viability (6.18 Log CFU/mL) after passage throughout simulated digestion in the passion fruit juice matrix. Both juices maintained high probiotic counts (>8.0 Log CFU/mL) during storage. Also, the yellow color was stable after 28 days of storage. Volatile compounds of passion fruit juices were modified after the fermentation process, such as ketones and alcohol formation degradation. The sensory profile of passion fruit juice was modified by single and co-culture fermentations. The fermented samples were mainly correlated with the terminologies "salty, acidic and bitter tastes" and "sweetener aftertaste". Overall, passion fruit juice proved to be an adequate food matrix to deliver the evaluated strains. However, individual strains or strain-strain interactions with the food matrix affect the fermented product, demonstrating that strain and matrices evaluations are essential for developing novel products with acceptable characteristics.

Cross-cultural adaptation and validation of the Urinary Incontinence Scale After Radical Prostatectomy for the Brazilian context.

OBJECTIVE: To translate, adapt, and validate the Urinary Incontinence Scale After Radical Prostatectomy for Brazil. METHOD: Methodological study: cross-cultural adaptation (translation, synthesis, back translation, expert committee (n=25), pre-testing (n=40) and presentation to original authors) and evaluation of measurement properties (n=80). Data were collected between January 2018 and February 2019 in an oncology unit. The calculated measurement properties: structural validity, hypothesis testing, criterion validity and reliability. RESULTS: The Brazilian version was called Escala de Incontinência Urinária Pós-Prostatectomia Radical. One item was excluded due to low factor loading (0.322). A significant correlation was identified between the total score of the scale and instruments applied (p<0.001). Incontinent men had higher scores on the total scale in relation to continents (p<0.001). Cronbach's alpha was 0.94 and composite reliability was 0.97. CONCLUSION: The Brazilian version was considered valid and reliable for the assessment of urinary incontinence in prostatectomized patients.

Evaluation of potentially probiotic yeasts and Lactiplantibacillus plantarum in co-culture for the elaboration of a functional plant-based fermented beverage.

This work aimed to evaluate the performance of co-cultivation of potential probiotic yeast and lactic acid bacteria (LAB) in producing plant-based fermented beverages. The co-culture comprised LAB Lactiplantibacillus plantarum CCMA0743 with the yeasts Pichia kluyveri CCMA 0615, Pichia guilliermondii CCMA 1753 and Debaryomyces hansenii CCMA 1761 separately. The plant substrate was 75 g oat, 175 g sunflower seeds, and 75 g almonds. The viability of microorganisms in the plant-based matrix was evaluated during fermentation, storage at 4 °C, and under simulated gastrointestinal tract (GIT) conditions. Chemical analysis, antioxidant activity, and sensory profile of the beverages were also determined. The three yeasts and the LAB showed counts greater than 6.0 log CFU/mL after fermentation, and the plant-based matrix protected the yeasts during simulated digestion. P. kluyveri and D. hansenii showed higher survival than P. guilliermondii and L. plantarum after exposure to simulated GIT conditions. The pH of the plant-based matrix reduced from approximately 7 to 3.8. Lactic acid was the main organic acid produced during fermentation. In addition, 113 volatile compounds were detected by gas chromatography-mass spectrometry (GC-MS), including alcohols, aldehydes, alkanes, alkenes, acids, ester, ether, ketones, phenol, and amides. The beverage sensory profile varied with the co-culture. The co-culture D. hansenii and L. plantarum showed higher antioxidant activity than the other co-culture tested, and the homogeneous texture attribute characterized the beverage produced with this combination. Results show the suitability of tested co-cultures to produce a plant-based fermented beverage and indicate more significant potential for D. hansenii and L. plantarum co-culture as a starter for its functionalization.

Interruption in the work of nursing professionals: conceptual analysis.

OBJECTIVES: to analyze the concept of "interruption in the work of nursing professionals". METHODS: conceptual study according to the method proposed by Walker and Avant through integrative literature review. The study searched the databases using the descriptors: "Attention", "Attention Bias", "Health Personnel", "Nurses", "Patient Safety" and "Medical Malpractices". The sample consisted of 36 studies. RESULTS: the antecedents were alarms, answering phone calls, providing patient care, and lack of material/medicine. The defining attributes were pause, suspension, breakage, and intrusion. In relation to the consequences, the study highlighted the increase in frequency and severity of medication errors and change of focus. FINAL CONSIDERATIONS: theconceptual analysis identified the attributes, antecedents, and consequences and allowed to build an operational definition for "interruption in the work of nursing professionals". It will contribute to the improvement of the work process and the creation of strategies that ensure safer care for the patient.

Probiotic Potential, Antioxidant Activity, and Phytase Production of Indigenous Yeasts Isolated from Indigenous Fermented Foods.

While many bacteria have been used as probiotics by industries, only two yeasts, Saccharomyces cerevisiae var. boulardii and Kluyveromyces fragilis (B0399), have been used for this purpose. In the present work, a total of 116 yeasts isolated from Brazilian indigenous fermented food, cocoa fermentation, and kefir were in vitro characterized for probiotic attributes. From 116 isolates, 36 were tolerant to gastrointestinal conditions evaluated by tolerance to pH 2.0, bile salts (0.3% w/v), and 37 °C temperature. From those, 15 isolates showed a similar or higher percentage (P < 0.05) of hydrophobicity, autoaggregation, and coaggregation with E. coli than the reference strain S. boulardii. All these strains showed a high percentage of adhesion to Caco-2 cells (> 63%) and antioxidant activity (ranging from 18 to 62%). Phytate hydrolysis was evaluated for these yeasts and 13 strains showed positive results, which is important for nutrient availability in plant-based foods. These results are important insights for characterization of novel probiotic yeast strains as well as to aggregate functional value to these food products.

Latino Students Patient Safety Questionnaire: cross-cultural adaptation for Brazilian nursing and medical students.

OBJECTIVES: to perform a cross-cultural adaptation of the Latino Student Patient Safety Questionnaire for Brazilian Nursing and Medical students. METHODS: methodological study carried out in six stages - forward translation, synthesis, back-translation, expert committee assessment, pre-test and reporting to the authors of the original instrument. Data at the expert committee and pre-test stages were collected and analyzed using the web platform e-Surv. The level of agreement adopted for the expert committee evaluation with 20 participants and the pre-test evaluation with 38 students was ≥ 90%. RESULTS: the first evaluation round by the expert committee showed a <90% agreement for 21 out of the 26 questionnaire items, requiring adjustments. In the pre-test, three items in the instrument reached a <90% agreement and were revised to obtain the final version. CONCLUSIONS: the Brazilian version of the Latino Student Patient Safety Questionnaire instrument was considered culturally adapted to Brazilian Portuguese.

Ion torrent-based nasopharyngeal swab metatranscriptomics in COVID-19.

Herein, we describe the detection of a SARS-CoV-2 genome through metatranscriptome next-generation sequencing directly from the nasopharyngeal swab of a suspected case of local transmission of Covid-19, in Brazil. Depletion of human ribosomal RNA and use of an optimized in-house developed bioinformatics strategy contributed to successful detection of the virus.

Complete genome sequence of human T-cell lymphotropic type 1 from patients with different clinical profiles, including infective dermatitis.

The HTLV-1 is the first human retrovirus and is associated with several clinical syndromes, however, the pathogenesis of these clinical manifestations is still not fully understood. Furthermore, there are few complete genomes publicly available, about 0.12 complete genomes per 10,000 infected individuals and the databases have a major deficiency of sequences information. This study generated and characterized 31 HTLV-1 complete genomes sequences derived from individuals with Tropical Spastic Paraparesis/HTLV-1-Associated Myelopathy (TSP/HAM), Adult T-cell leukemia/lymphoma (ATL), infective dermatitis associated to HTLV-1 (IDH) and asymptomatic patients. These sequences are associated to clinical and epidemiological information about the patients. The sequencing data generated on Ion Torrent PGM platform were assembled and mapped against the reference HTLV-1 genome. These sequences were genotyped as Cosmopolitan subtype, Transcontinental subgroup. We identified the variants in the coding regions of the genome of the different clinical profiles, however, no statistical relation was detected. This study contributed to increase of HTLV-1 complete genomes in the world. Furthermore, to better investigate the contribution of HTLV-1 mutations for the disease outcome it is necessary to evaluate the interaction of the viral genome and characteristics of the human host.

Prostaglandin mediates endotoxaemia-induced hypophagia by activation of pro-opiomelanocortin and corticotrophin-releasing factor neurons in rats.

Corticotrophin-releasing factor (CRF) and alpha-melanocyte-stimulating hormone (alpha-MSH), both of which are synthesized by hypothalamic neurons, play an essential role in the control of energy homeostasis. Neuroendocrine and behavioural responses induced by lipopolyssacharide (LPS) have been shown to involve prostaglandin-mediated pathways. This study investigated the effects of prostaglandin on CRF and alpha-MSH neuronal activities in LPS-induced anorexia. Male Wistar rats were pretreated with indomethacin (10 mg kg(-1); i.p.) or vehicle; 15 min later they received LPS (500 microg kg(-1); i.p.) or saline injection. Food intake, hormone responses and Fos-CRF and Fos-alpha-MSH immunoreactivity in the paraventricular and arcuate nuclei, respectively, were evaluated. In comparison with saline treatment, LPS administration induced lower food intake and increased plasma ACTH and corticosterone levels, as well as an increase in Fos-CRF and Fos-alpha-MSH double-labelled neurons in vehicle-pretreated rats. In contrast, indomethacin treatment partly reversed the hypophagic effect, blunted the hormonal increase and blocked the Fos-CRF and Fos-alpha-MSH hypothalamic double labelling increase in response to the LPS stimulus. These data demonstrate that the activation of pro-opiomelanocortin and CRF hypothalamic neurons following LPS administration is at least partly mediated by the prostaglandin pathway and is likely to be involved in the modulation of feeding behaviour during endotoxaemia.

Live Cell Imaging Supports a Key Role for Histone Deacetylase as a Molecular Target during Glioblastoma Malignancy Downgrade through Tumor Competence Modulation.

Glioblastoma (GBM) is the most aggressive tumor of the central nervous system, and the identification of the mechanisms underlying the biological basis of GBM aggressiveness is essential to develop new therapies. Due to the low prognosis of GBM treatment, different clinical studies are in course to test the use of histone deacetylase inhibitors (iHDACs) in anticancer cocktails. Here, we seek to investigate the impact of HDAC activity on GBM cell behavior and plasticity by live cell imaging. We pharmacologically knock down HDAC activity using two different inhibitors (TSA and SAHA) in two different tumor cell types: a commercial GBM cell line (U87-MG) and primary tumor (GBM011). Upon 72 hours of in vitro iHDAC treatment, GBM cells presented a very unusual elongated cell shape due to tunneling tube formation and independent on TGF-ß signaling epithelial to mesenchymal transition. Live cell imaging revealed that voltage-sensitive Ca++ signaling was disrupted upon HDAC activity blockade. This behavior was coupled to vimentin and connexin 43 gene expression downregulation, suggesting that HDAC activity blockade downgrades GBM aggressiveness mostly due to tumor cell competence and plasticity modulation in vitro. To test this hypothesis and access whether iHDACs would modulate tumor cell behavior and plasticity to properly respond to environmental cues in vivo, we xenografted GBM oncospheres in the chick developing the neural tube. Remarkably, upon 5 days in the developing neural tube, iHDAC-treated GBM cells ectopically expressed HNK-1, a tumor-suppressor marker tightly correlated to increased survivor of patients. These results describe, for the first time in the literature, the relevance of iHDACs for in vivo tumor cell morphology and competence to properly respond to environmental cues. Ultimately, our results highlight the relevance of chromatin remodeling for tumor cell plasticity and shed light on clinical perspectives aiming the epigenome as a relevant therapeutic target for GBM therapy.

Prevalence of intestinal parasites in children from public daycare centers in the city of Belo Horizonte, Minas Gerais, Brazil.

The objective of this study was to verify the occurrence of intestinal parasites in 3 to 6-year-old children from daycare centers maintained by the municipal government of Belo Horizonte, Minas Gerais, Brazil. Coproparasitological tests performed in 472 children have shown that 24.6% of them had some type of parasites, 6.6% of the children having more than one type. Among protozoa, Entamoeba coli (14.0%) and G. duodenalis (9.5%) were the most prevalent, whereas Ascaris lumbricoides (3.0%) and Trichuris trichiura (1.1%) were the most frequent among the helminths. Thus, we can observe that intestinal parasites still represent a serious public health problem in Belo Horizonte, especially among children and in areas where the socioeconomic conditions are less favorable.

Combination of probiotic yeast and lactic acid bacteria as starter culture to produce maize-based beverages.

Cereal-based fermented beverages are non-dairy products which are considered possible carriers for probiotic strains and alternatives for use by vegans and lactose-intolerant consumers. In the present work, the commercial probiotic, Lactobacillus paracasei LBC-81, was used singly and in co-culture with potential probiotic yeasts, Saccharomyces cerevisiae CCMA 0731, S. cerevisiae CCMA 0732, and Pichia kluyveri CCMA 0615, to ferment a maize-based substrate. All tested strains showed viability higher than 6 log CFU/mL, as recommended for food probiotic products, except for the yeast P. kluyveri which decreased during fermentation and storage time. A reduction in pH value, from approximately 7 to 4, was observed. This decrease was due organic acid production, which did not affect the microbial viability. Lactic and acetic acids were the main organic acids produced during fermentation, and they decreased over 28 days of storage (<0.5 and 0.1 g/L for lactic and acetic acids, respectively). Ethanol was detected in the S. cerevisiae assays; however, the content was <5 g/L in a non-alcoholic beverage. Seventy volatile compounds were detected, including acids, alcohols, aldehydes, esters, ketones, and other compounds. Sensory analysis showed score of 5.93-4.57, respectively for appearance and taste. This is an important result, considering that the beverage had no flavoring additive and lacked a sweet taste. Therefore, probiotic beverages were successfully obtained by maize fermentation inoculated with co-culture of S. cerevisiae (CCMA 0731 or CCMA 0732) and L. paracasei LBC-81.