La caracterización fenotípica y genotípica de Serratia marcescens proveniente de la Unidad de Neonatología de Referencia de Belém (Pará, Brasil)

Centro Universitário do Pará. Curso de Pós-Graduação em Análises Clínicas. Belém, PA, Brasil / Laboratório de Patologia Clínica e Laboratorial Dr. Paulo Azevedo. Belém, PA, Brasil.Universidade Federal do Pará. Departamento de Medicina. Belém, PA, Brasil.Centro Universitário do Pará. Curso de Pós-Graduação em Análises Clínicas. Belém, PA, Brasil.Laboratório de Patologia Clínica e Laboratorial Dr. Paulo Azevedo. Belém, PA, Brasil.Laboratório de Patologia Clínica e Laboratorial Dr. Paulo Azevedo. Belém, PA, Brasil.Centro Universitário do Pará. Curso de Pós-Graduação em Análises Clínicas. Belém, PA, Brasil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.A Serratia marcescens tem sido relatada como importante agente de infecções relacionadas à saúde, destacando-se por apresentar elevado nível de resistência intrínseca aos antimicrobianos usados em neonatologia, além de persistir por longos períodos no ambiente hospitalar. Neste trabalho foram avaliadas, por métodos fenotípicos e moleculares, S. marcescens recuperadas a partir de colonização do trato gastrointestinal ou sepse tardia em neonatos internados em Unidade Neonatal em Belém. A identificação das S. marcescens e o teste de sensibilidade foram realizados por meio de sistema automatizado Vitek (BioMérieux); a suscetibilidade ao ertapenem foi avaliada com auxílio de disco contendo 10 µg da droga (Oxoide). A genotipagem foi feita por ERIC-PCR usando os primers ERIC1 (5'-TGAATCCCCAGGAGCTTACAT-3') e ERIC2 (5'-AAGTAAGTGACTGGGGTGAGCG-3'). Foram obtidas 22 cepas de S. marcescens, sendo 15 recuperadas de hemoculturas, e sete de vigilância (swab retal); todas apresentaram resistência a: ampicilina, ampicilina-sulbactam, gentamicina e cefalotina. Não foi observada resistência a: ciprofloxacina, imipenem, meropenem e ertapenem. Quanto aos demais antibióticos avaliados, o perfil de suscetibilidade foi variável. Foram obtidos 11 padrões de amplificação por ERIC-PCR, dois foram compartilhados por 14 isolados. Foi possível observar um padrão polimórfico característico para as cepas provenientes de colonização gastrointestinal, exceto em dois casos, que apresentaram padrões genotípicos relacionadas a casos de sepse. Os dados obtidos neste trabalho confirmam o elevado índice de resistência da S. marcescens aos antimicrobianos; no entanto, todos os isolados apresentaram sensibilidade à ciprofloxacina e aos carbapenêmicos. A tipagem por meio de antibiograma e ERIC-PCR sugere dispersão de clones associados à colonização ou sepse entre alas na Unidade Neonatal do hospital estudado.Serratia marcescens has been reported as an important agent of health care-related infections and has been highlighted for presenting a high level of intrinsic resistance to antimicrobials used in neonatology, besides persisting in hospital environments for long periods. In this work, S. marcescens was recovered from colonies in the gastrointestinal tract or late sepsis in newborn infants hospitalized in a Neonatal Unit in Belém. The identification of S. marcescens and the sensitivity test was carried out using a Vitek (BioMérieux) automated system; susceptibility to ertapenem was assessed using e-test strips (Oxoid). Genotyping was executed by ERIC-PCR using the primers ERIC1 (5’-TGAATCCCCAGGAGCTTACAT-3’) and ERIC2 (5’-AAGTAAGTGACTGGGGTGAGCG-3’). Twenty-two strains of S. marcescens were recovered: 15 from hemocultures and seven from surveillance (rectal swab culture). All presented resistance to ampicillin, ampicillinsulbactam, gentamicin and cephalothin. There were no indications of resistance to ciprofloxacin, imipenem, meropenem or ertapenem. The susceptibility profiles varied for other antibiotics. Eleven amplification patterns by ERIC-PCR were obtained, and two were shared by 14 isolates. It was possible to observe a characteristic polymorphic pattern in the strains from gastrointestinal colonization, except for two cases, which presented genotypic patterns related to cases of sepsis. The data obtained in this work confirm the high level of resistance of S. marcescens against antimicrobials; however, all isolates displayed sensitivity to ciprofloxacin and carbapenemics. Antibiogram and ERIC-PCR typing suggest a dispersion of clones associated with colonization or sepsis among the wards of the Neonatal Unit in the surveyed hospital

α-Tocopherol influences glycaemic control and miR-9-3 DNA methylation in overweight and obese women under an energy-restricted diet: a randomized, double-blind, exploratory, controlled clinical trial

Abstract Background Excess weight is a strong risk factor for the development of dysglycaemia. It has been suggested that changes in the metabolism microRNAs, small non-coding RNAs that regulate gene expression, could precede late glycaemic changes. Vitamin E in turn may exert important functions in methylation and gene expression processes. This study aimed to determine the effect of α-tocopherol on glycaemic variables and miR-9-1 and miR-9-3 promoter DNA methylation in overweight women. Methods A randomized, double-blind, exploratory, placebo-controlled study was conducted in overweight and obese adult women (n = 44) who ingested synthetic vitamin E (all-rac-α-tocopherol), natural source vitamin E (RRR-rac-α-tocopherol) or placebo capsules and were followed up for a period of 8 weeks. Supplemented groups also received dietary guidance for an energy-restricted diet. An additional group that received no supplementation and did not follow an energy-restricted diet was also followed up. The intervention effect was evaluated by DNA methylation levels (quantitative real-time PCR assay) and anthropometric and biochemical variables (fasting plasma glucose, haemoglobin A1C, insulin, and vitamin E). Results Increased methylation levels of the miR-9-3 promoter region (P < 0.001) and reduced haemoglobin A1C (P < 0.05) were observed in the natural source vitamin E group after intervention. Increased fasting plasma glucose was observed in the synthetic vitamin E group, despite the significant reduction of anthropometric variables compared to the other groups. Conclusions α-Tocopherol from natural sources increased methylation levels of the miR-9-3 promoter region and reduced haemoglobin A1C in overweight women following an energy-restricted diet. These results provide novel information about the influence of vitamin E on DNA methylation. Trial registration ClinicalTrials.gov, NCT02922491. Registered 4 October, 2016

Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults

Abstract Background DNA methylation has been evidenced as a potential epigenetic mechanism related to various candidate genes to development of obesity. Therefore, the objective of this study was to evaluate the DNA methylation levels of the ADRB3 gene by body mass index (BMI) in a representative adult population, besides characterizing this population as to the lipid profile, oxidative stress and food intake. Methods This was a cross-sectional population-based study, involving 262 adults aged 20–59 years, of both genders, representative of the East and West regions of the municipality of João Pessoa, Paraíba state, Brazil, in that were evaluated lifestyle variables and performed nutritional, biochemical evaluation and DNA methylation levels of the ADRB3 gene using high resolution melting method. The relationship between the study variables was performed using analyses of variance and multiple regression models. All results were obtained using the software R, 3.3.2. Results From the stratification of categories BMI, was observed a difference in the average variables values of age, waist-to-height ratio, waist-to-hip ratio, waist circumference, triglycerides and intake of trans fat, which occurred more frequently between the categories “eutrophic” and “obesity”. From the multiple regression analysis in the group of eutrophic adults, it was observed a negative relationship between methylation levels of the ADRB3 gene with serum levels of folic acid. However, no significant relation was observed among lipid profile, oxidative stress and food intake in individuals distributed in the three categories of BMI. Conclusions A negative relationship was demonstrated between methylation levels of the ADRB3 gene in eutrophic adults individuals with serum levels of folic acid, as well as with the independent gender of BMI, however, was not observed relation with lipid profile, oxidative stress and variables of food intake. Regarding the absence of relationship with methylation levels of the ADRB3 gene in the categories of overweight, mild and moderate obesity, the answer probably lies in the insufficient amount of body fat to initiate inflammatory processes and oxidative stress with a direct impact on methylation levels, what is differently is found most of the times in exacerbated levels in severe obesity