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Infection by Adenovirus 36 (Ad-36) has been associated with adipogenesis using cell and animal models, and a high risk of developing obesity has been reported in Ad-36-seropositive individuals. However, molecular mechanisms involved in the maintenance over the years of adipogenesis associated with Ad-36 has not been investigated in human adipose tissue. Epigenetic mechanisms, such as micro-RNAs (miRNAs) that regulate gene expression at the post-transcriptional level, have shown an important role in the development and maintenance of metabolic diseases. This study investigated the expression of miRNA associated with the adipogenic process in visceral adipose tissue from obese individuals according to Ad-36 serology. Obese individuals were separated according to their status of Ad-36 serology in seropositive (Ad-36 (+); n = 29) and seronegative (Ad-36 (−); n = 28) groups. Additionally, a group of lean controls (n = 17) was selected to compare with obese individuals. Biopsies of visceral adipose tissue were obtained to evaluate miRNA and gene expression. The study of Ad-36 serology was carried out by ELISA. The expression of pro-adipogenic (miR-17 and miR-210) and anti-adipogenic (miR-155, miR-130 and miR-27a) miRNAs was evaluated using Taqman advanced miRNA assays by qPCR. The expression of adipogenes encoding LEP, ADIPOQ, and PPARγ was evaluated by Taqman predesigned assays through qPCR. The obese group had higher LEP (p

Helicobacter pylori is the main bacteria associated with gastroduodenal diseases. Recent studies have reported that gastric microbiota might be modified by the H. pylori colonization, favoring gastric lesions&prime, development. In Chile, the region of La Araucan&iacute, a concentrates a high risk of gastric cancer associated with Helicobacter pylori colonization, rurality, poverty, and Mapuche ethnicity. Hence, we aimed to identify the culturable gastric microbiota and characterize its variability at different stages of epithelial injury, based on its H. pylori colonization in dyspeptic patients from this Chilean region. Microaerophilic bacteria strains were isolated from antrum biopsies of 155 dyspeptic patients&prime, biopsies and identified using MALDI-TOF MS or 16sRNA gene sequencing for non-pylori species identification, and UreC gene amplification for H. pylori confirmation. We found 48 species from 18 families, mainly belonging to Neisseriaceae (21.3%), Streptococcaceae (20.0%), Actynomicetaceae (9.0%), Enterobacteriaceae, and Lactobacillaceae (4.5%), however, Streptococcaceae and Actinomycetaceae families showed a significant reduction in samples infected with H. pylori, along with a considerably lower diversity of species. Our results revealed a microbiota modification due to H. pylori colonization associated with the gastric epithelial state, suggesting a potential microbiota role for developing and progressing gastric diseases.

Adenovirus 36 (Ad-36) has been associated to adiposity in animal and in vitro studies. Ad-36 seropositivity has also been reported to contribute to obesity risk in children and adult populations. We investigated the relationship of Ad-36 serology with obesity and metabolic parameters in a Chilean population. Clinical and anthropometric data were obtained and blood samples were drawn from 99 lean (BMI: 18.5–24.9 kg/m2) and 151 obese (BMI > 30 kg/m2) subjects. Laboratory tests included lipid profile as well as glucose, insulin, leptin, and adiponectin levels. Ad-36 seropositivity was evaluated in serum samples by enzyme-linked immunosorbent assay. Seroprevalence of Ad-36 was higher in the obese group (58%) than in lean controls (34%) demonstrating that individuals previously infected with Ad-36 have higher risk of obesity in the study population (OR: 2.67, 95%CI: 1.58–4.51, p

Helicobacter pylori colonizes half of the human population. Age, ethnicity, and socioeconomic status are factors that influence the prevalence of the infection. This is important in southern Chile, one of the most unequal regions in the world, where a significant difference in the health access of the population occurs due to the existence of two competing health systems. Moreover, in the last few years, current protocols of H. pylori eradication have shown high rates of resistance with reduced therapeutic efficacy. This study reported the epidemiology of infection and attempted to identify divergent points among the population beneficiaries of the two health care schemes in southern Chile. Biopsies from public (n = 143) and private (n = 86) health systems were studied. At the same time, clinical and sociodemographic factors were evaluated. H. pylori strains were obtained from gastric biopsies for culture and molecular testing. Antibiotic susceptibility was determined by the agar dilution method. Differences about ethnicity, rural residence, and education (p &le, 0.05) were observed between beneficiaries of the two health systems. The prevalence of H. pylori was 45%, with no significant differences regardless of the socioeconomic conditions. The only identified risk factor associated with H. pylori infection was Mapuche ethnicity (OR (odds ratio) = 2.30). H. pylori showed high resistance rates, particularly against clarithromycin (40%), levofloxacin (43.1%), and metronidazole (81.8%). This study highlighted the importance of Mapuche ancestry as a risk factor in southern Chile and emphasized the need to search for new eradication strategies as well as further studies evaluating therapeutic efficacy.

Enterobacteria-producing extended-spectrum β-lactamases (ESBL) play an important role in healthcare infections, increasing hospitalization time, morbidity and mortality rates. Among several ESBLs that emerge from these pathogens, CTX-M-type enzymes had the most successful global spread in different epidemiological settings. Latin America presents high prevalence of CTX-M-2 in ESBL-producing enterobacterial infections with local emergence of the CTX-M-1 group. However, this high prevalence of the CTX-M-1 group has not yet been reported in Chile. The aim of this study was to identify ESBLs among enterobacteria isolated from clinical samples of critically ill patients from southern Chile. One-hundred thirty seven ESBL-producing bacteria were isolated from outpatients from all critical patient units from Hernán Henríquez Aravena Hospital. Phenotype characterization was performed by antibiogram, screening of ESBL, and determination of minimum inhibitory concentration (MIC). PCR was used for genetic confirmation of resistance. Molecular typing was performed by ERIC-PCR. ESBL-producing isolates were identified as Klebsiella pneumoniae (n = 115), Escherichia coli (n = 18), Proteus mirabilis (n = 3), and Enterobacter cloacae (n = 1), presenting multidrug resistance profiles. PCR amplification showed that the strains were positive for blaSHV (n = 111/81%), blaCTX-M-1 (n = 116/84.7%), blaTEM (n = 100/73%), blaCTX-M-2 (n = 28/20.4%), blaCTX-M-9 (0.7%), blaPER-1 (0.7%), and blaGES-10 (0.7%). The multiple production of ESBL was observed in 93% of isolates, suggesting high genetic mobility independent of the clonal relationship. The high frequency of the CTX-M-1 group and a high rate of ESBL co-production are changing the epidemiology of the ESBL profile in Chilean intensive care units. This epidemiology is a constant and increasing challenge, not only in Chile, but worldwide. Keywords: ESBL-producing enterobacteria, Intensive care units, CTX-M-1 group

The development of novel biological products, such as enzymes, dyes, antibiotics, and others with high potential use for biotechnological applications, continues to stimulate the search for microorganisms in extreme environments. In recent years, the study of Antarctic ecosystems and their microorganisms has received more attention because psychrophilic microorganisms—mainly bacteria and their enzymes—have assumed considerable importance for biotechnological applications. In this study, we characterized a collection of 30 culturable Actinobacteria isolated from the South Shetland Islands, Antarctica, in order to relate their distribution and ability to produce extracellular enzymes with their biotechnological potential. The isolates belonged to eight genus in the phylum Actinobacteria: Arthrobacter, Brevibacterium, Curtobacterium, Janibacter, Knoellia, Rhodococcus, Streptomyces and Thermoleophilum. The screening for biologically active products showed that all the evaluated strains produced at least one extracellular enzyme at 4 °C, with protease, gelatinase and cellulase enzymes being the most common. In conclusion, our data showed that various Antarctic ecosystems are good sources for isolating cultivable Actinobacteria. Furthermore, these bacteria provide high-quality biological material for screening cold-active enzymes that have potential biotechnological applications and economic benefits.

INTRODUCTION Clopidogrel is commonly used in prevention and treatment of atherothrombosis. Some previous studies have suggested a pleiotropic effect of clopidogrel; however, when this drug causes platelet-independent effects on endothelial function remains unclear. AIMS To evaluate the influence of clopidogrel on inflammatory biomarkers and adhesion molecules in human endothelial cells and the role of nitric oxide (NO) in this process. METHODS TNF-α-induced human umbilical vein endothelial cells (HUVEC) were exposed to clopidogrel. Gene expression and protein expression of ICAM-1, P-selectin, IL-8, IL-6, and MCP-1 were evaluated by qPCR, flux cytometry, or milliplex technology. Expression of endothelial nitric oxide synthase (NOS3) and NO release were also evaluated. Influence of clopidogrel was further evaluated in NOS3 downregulated HUVEC by RNAi. RESULTS Clopidogrel at 20 μmol/L induced NO release in HUVEC after 24-hours treatment. Gene expressions of inflammatory markers IL-8 and MCP1 were reduced after clopidogrel treatment (P

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brazil / Grupo Fleury. S?o Paulo, SP, Brazil. Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Laborat?rio de Tecnologia Biomolecular. Bel?m, Par?, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brasil. Universidade Estadual de Campinas. Faculdade de Ci?ncias M?dicas. Departamento de Patologia Cl?nica. Campinas, SP, Brazil. Universidade de S?o Paulo. Faculdade de Ci?ncias Farmac?uticas. Departamento de An?lises Cl?nicas e Toxicol?gicas. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Faculdade de Ci?ncias Farmac?uticas. Departamento de An?lises Cl?nicas e Toxicol?gicas. S?o Paulo, SP, Brazil / Universidade de S?o Paulo. Instituto de Ci?ncias Biom?dicas. Departamento de Microbiologia. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brasil / Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Laborat?rio de Tecnologia Biomolecular. Bel?m, Par?, Brazil. Universidade de S?o Paulo. Faculdade de Ci?ncias Farmac?uticas. Departamento de An?lises Cl?nicas e Toxicol?gicas. S?o Paulo, SP, Brazil. Grupo Fleury. S?o Paulo, SP, Brazil / Universidade de S?o Paulo. Faculdade de Ci?ncias Farmac?uticas. Departamento de An?lises Cl?nicas e Toxicol?gicas. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro de Inova??es Tecnol?gicas. Ananindeua, PA, Brasil / Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Laborat?rio de Tecnologia Biomolecular. Bel?m, Par?, Brazil. We sequenced the oldest blaKPC-2-bearing plasmid isolated in Brazil and another plasmid also carried by a Klebsiella pneumoniae strain of sequence type 442 (ST442), isolated 52 months later. Both plasmids present an IncN backbone and few acquired regions. Because the 2005 plasmid presented deletions and a truncated gene within Tn4401b compared to the 2009 plasmid, we can thus infer that IncN blaKPC-2-bearing plasmids pFCF1305 and pFCF3SP had a common ancestor circulating in Brazil prior toMay 2005.

Intrinsic mechanisms leading to carbapenem-induced membrane impermeability and multidrug resistance are poorly understood in clinical isolates of Enterobacteriaceae. In this study, molecular behaviours during the establishment of membrane impermeability in members of the Enterobacteriaceae family under imipenem selective pressure were investigated. Clinical isolates (n = 22) exhibiting susceptibility to multiple antibiotics or characterised as extended-spectrum β-lactamase (ESBL)- or AmpC-producers were submitted to progressive passages on Mueller-Hinton agar plates containing subclinical concentrations of imipenem [0.5 × the minimum inhibitory concentration (MIC)]. Changes in outer membrane permeability were evaluated by determination of antimicrobial MICs, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and gene expression analysis related to membrane permeability (i.e. omp35-like, omp36-like and acrA) and regulatory mechanisms (i.e. marA and ompR) by quantitative reverse transcription PCR. Following imipenem induction, 73% of isolates showed increased carbapenem MICs by ≥2 doubling dilutions. At an early stage of treatment, imipenem modulated the expression of porins and efflux pump genes, represented by a reduction of 78% in omp36-like and a two-fold increase in acrA expression. Transcriptional factors marA and ompR were also affected by imipenem induction, increasing mRNA expression by 14- and 4-fold, respectively. High marA expression levels were associated with higher values of acrA expression. These results suggest that imipenem is an important factor in the development of an adaptive response to carbapenems by regulating key genes involved in the control of efflux pumps and porins, which could lead to a multidrug-resistant profile in clinical isolates, contributing to possible treatment failure.

Introduction: Linezolid has excellent activity against multidrug‐resistant Staphylococcus spp. and linezolid resistance has emerged mainly in isolates recovered from patients who are receiving courses of oxazolidinone therapy. Case presentation: We report a clinical case of bacteraemia caused by a linezolid‐resistant Staphylococcus epidermidis clone ST‐2 with the G2576T mutation in five copies of the 23S rRNA gene (MIC, 32 µg ml−1) isolated from a patient who did not receive any course of oxazolidinone therapy. The S. epidermidis HSPE24‐SP strain was WT for L3, L4 and L22 ribosomal proteins, cfr negative and indistinguishable by PFGE from the clone that circulated at the same time in another tertiary care hospital located in the same city. Conclusion: This report suggests the inter‐hospital spread of S. epidermidis clone ST‐2 and calls attention to the possibility of isolation of a linezolid‐resistant strain with multiple 23S rRNA mutated copies in an environment under no oxazolidinone selective pressure.

Linezolid resistance in vancomycin-resistant enterococci (VRE) strains has been rarely reported, with Enterococcus faecium being the species most commonly associated with these few cases ([1][1], [2][2], [3][3]). Here, we report infections due to linezolid- and vancomycin-resistant Enterococcus (

Department of Microbiology, Institute of Biomedical Sciences, Universidade de Sao Paulo, Sao Paulo, SP, Brazil; Department of Clinical Analysis, School of Pharmacy, Universidade de Sao Paulo, Sao Paulo, SP, Brazil; Institute of Biological Sciences, Universidade Federal do Para, Belem, PA, Brazil; School of Public Health, Universidade de Sao Paulo, Sao Paulo, SP, Brazil; Environmental Company of Sao Paulo State (CETESB), Sao Paulo, SP, Brazil

A recent publication by Monteiro et al. reported the first detection of KPC-2 β-lactamase in carbapenem-resistant Klebsiella pneumoniae strains in Brazil, which occurred in 2006 ([5][1]). The authors reported that, between September and November of 2006, four carbapenem-resistant K. pneumoniae

Among different clinical staphylococcal strains, linezolid resistance has been mainly mediated by mutations in the central loop of domain V of 23S rRNA, with the G2576T mutation being the most prevalent.…