RESUMO
Helicobacter pylori (H. pylori) induces an intense inflammatory response, mediated by proinflammatory cytokines, including interleukin (IL)-6 and its membrane receptor (IL-6R), which activates important signaling pathways in the development of gastric disease and cancer. We investigated the gene and protein expression of IL-6 and IL-6R and the influence of polymorphisms rs1800795, rs1800796, and rs1800797 on its gene expression together with H. pylori infection. Furthermore, an in-silico analysis was performed to support our results. Gastric biopsies were obtained from patients with gastric symptoms and patients with gastric cancer (GC) and were divided into groups (Control, Gastritis, and Cancer). H. pylori was detected by PCR. Real-time-qPCR was employed to determine gene expression, and western blot assay was used to analyze protein expression levels. PCR-RFLP was used to characterize IL-6 polymorphisms. Bioinformatics analyses were performed using the Gene Expression Omnibus (GEO) database and GEO2R to screen out differentially expressed genes (DEGs). H. pylori was detected in 43.3% of the samples. Statistically significant differences were found for IL-6 (P=0.0001) and IL-6R (P=0.0005) genes among the three groups, regardless of the presence of H. pylori. Among patients with H. pylori infection, the IL-6 and IL-6R gene and protein expressions were significantly increased, highlighting IL-6 gene overexpression in patients with GC. No statistically significant differences were found for the rs1800795, rs1800796, and rs1800797 polymorphisms compared to IL-6 gene expression. The results indicated that the IL-6 polymorphisms do not influence its expression, but IL-6 and IL-6R expression seems to be altered by the presence of H. pylori.
Assuntos
Gastrite , Infecções por Helicobacter , Helicobacter pylori , Interleucina-6/genética , Neoplasias Gástricas , Mucosa Gástrica , Gastrite/genética , Infecções por Helicobacter/genética , Humanos , Interleucina-8 , Neoplasias Gástricas/genéticaRESUMO
Helicobacter pylori (H. pylori) induces an intense inflammatory response, mediated by proinflammatory cytokines, including interleukin (IL)-6 and its membrane receptor (IL-6R), which activates important signaling pathways in the development of gastric disease and cancer. We investigated the gene and protein expression of IL-6 and IL-6R and the influence of polymorphisms rs1800795, rs1800796, and rs1800797 on its gene expression together with H. pylori infection. Furthermore, an in-silico analysis was performed to support our results. Gastric biopsies were obtained from patients with gastric symptoms and patients with gastric cancer (GC) and were divided into groups (Control, Gastritis, and Cancer). H. pylori was detected by PCR. Real-time-qPCR was employed to determine gene expression, and western blot assay was used to analyze protein expression levels. PCR-RFLP was used to characterize IL-6 polymorphisms. Bioinformatics analyses were performed using the Gene Expression Omnibus (GEO) database and GEO2R to screen out differentially expressed genes (DEGs). H. pylori was detected in 43.3% of the samples. Statistically significant differences were found for IL-6 (P=0.0001) and IL-6R (P=0.0005) genes among the three groups, regardless of the presence of H. pylori. Among patients with H. pylori infection, the IL-6 and IL-6R gene and protein expressions were significantly increased, highlighting IL-6 gene overexpression in patients with GC. No statistically significant differences were found for the rs1800795, rs1800796, and rs1800797 polymorphisms compared to IL-6 gene expression. The results indicated that the IL-6 polymorphisms do not influence its expression, but IL-6 and IL-6R expression seems to be altered by the presence of H. pylori.
Assuntos
Humanos , Neoplasias Gástricas/genética , Helicobacter pylori , Infecções por Helicobacter/genética , Interleucina-6/genética , Gastrite/genética , Interleucina-8 , Mucosa GástricaRESUMO
The EuroSIDA study was initiated in 1994 and follows adult people living with HIV (PLHIV) in 100 collaborating clinics across 35 countries covering all European regions, Israel and Argentina. The study aims to study the long-term virological, immunological and clinical outcomes of PLHIV and to monitor temporal changes and regional differences in outcomes across Europe. Annually collected data include basic demographic characteristics, information on AIDS- and non-AIDS-related clinical events, and details about antiretroviral therapy (ART), hepatitis C treatment and other medications, in addition to a range of laboratory values. The summer 2016 data set held data from a total of 23 071 individuals contributing 174 481 person-years of follow-up, while EuroSIDA's unique plasma repository held over 160 000 samples. Over the past 25 years, close to 300 articles have been published in peer-reviewed journals (h-index 52), covering a range of scientific focus areas, including monitoring of clinical and virological outcomes, ART uptake, efficacy and adverse events, the influence of hepatitis virus coinfection, variation in the quality of HIV care and management across settings and regions, and biomarker research. Recognizing that there remain unresolved issues in the clinical care and management of PLHIV in Europe, EuroSIDA was one of the cohorts to found The International Cohort Consortium of Infectious Disease (RESPOND) cohort consortium on infectious diseases in 2017. In celebration of the EuroSIDA study's 25th anniversary, this article aims to summarize key scientific findings and outline current and future scientific focus areas.
Assuntos
Infecções por HIV/tratamento farmacológico , HIV/imunologia , Hepatite C/tratamento farmacológico , RNA Viral/genética , Argentina , Contagem de Linfócito CD4 , Coinfecção , Europa (Continente) , Feminino , HIV/genética , Infecções por HIV/imunologia , Infecções por HIV/virologia , Humanos , Israel , Perda de Seguimento , Masculino , Estudos Multicêntricos como Assunto , Resultado do Tratamento , Carga ViralRESUMO
Alzheimer's Disease (AD) is the most common cause of dementia in elderly people. The presynaptic terminal is an important site of pathological changes in AD, leading to synaptic loss in specific brain regions, such as in the cortex and hippocampus. In this study, we investigated synaptosomal-associated protein, 25-kDa (SNAP25) mRNA levels and promoter DNA methylation in post mortem brain tissues (entorhinal and auditory cortices and hippocampus) from healthy elderly and AD subjects as well as in peripheral blood leukocytes of young, healthy elderly and AD patients. mRNA quantification was performed by quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) using the ΔΔC(T) method and promoter DNA methylation was quantified by mass spectrometry using the Sequenom EpiTYPER platform. We observed a significant decrease in SNAP25 expression in AD across all the three brain regions in relation to the healthy elderly subjects, suggesting impairment in synaptic function. The changes in the auditory cortex reflected those observed in the hippocampus and entorhinal cortex, the primary areas affected in AD. However, no AD-associated differences in SNAP25 promoter DNA methylation were observed suggesting that other mechanisms may be involved in mediating the observed gene expression changes.
Assuntos
Doença de Alzheimer/genética , Encéfalo/metabolismo , Metilação de DNA/genética , Regiões Promotoras Genéticas/genética , Proteína 25 Associada a Sinaptossoma/genética , Idoso , Doença de Alzheimer/metabolismo , Feminino , Humanos , Masculino , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real , Proteína 25 Associada a Sinaptossoma/biossíntese , Sinaptossomos/metabolismo , TranscriptomaRESUMO
Helicobacter pylori, a gram-negative bacterium, possesses two important virulence factors: the vacuolating toxin (vacA), and the cytotoxin-associated gene product (cagA). The aim of the present study was to evaluate the presence of H. pylori in the stomach and oral cavity of humans and compare the cagA and vacA genotypes of H. pylori found in different samples (stomach, saliva and dental plaque) from the same patient. Gastric biopsies, saliva and dental plaques were obtained from 62 dyspeptic adults. DNA was extracted and evaluated for the presence of H. pylori and the alleles cagA and vacA. Persons with gastritis had a higher frequency of H. pylori -positive samples in the stomach while positive samples from gastric biopsies were significantly correlated with those from the oral cavity. There was a high H. pylori frequency in patients while the cagA gene was associated with vacA s1 alleles in gastric biopsies. Our results suggest a reservoir of the species in the oral cavity and that, in one patient, more than one H. pylori strain may exist in the saliva, dental plaque and stomach. We found a relationship between gastric infection and the bacterium in the oral cavity, with the cytotoxin genotype varying between saliva and dental plaque.(AU)
Assuntos
Humanos , Helicobacter pylori/citologia , Dispepsia/diagnóstico , Dispepsia/microbiologia , Virulência , Saliva/microbiologia , Técnicas de Genotipagem/métodosRESUMO
The risk of developing gastric cancer is believed to be related to differences among Helicobacter pylori strains and the inflammatory responses mediated by host genetic factors. H. pylori infection is acquired at an early age and in the absence of appropriate antibiotic therapy, it generally persists for life. Tp53 gene regulates the transcription of several cytokines and chemokines involved in innate immunity and its action may be influenced by the presence of different H. pylori strains. The present study aimed to detect H. pylori in pediatric patients, to access Tp53 polymorphism at codon 72 and to correlate such findings with age and histopathological results. Three hundred and forty-two patients were analyzed. DNA from their gastric biopsies was extracted and the detection of H. pylori was performed through polymerase chain reaction assays, urease test and histopathologic examination. Allelic discrimination of SNP rs1042522 (Tp53) was performed by real-time polymerase chain reaction. Our results suggest a possible relationship between the presence of H. pylori and chronic gastritis in children and young patients, and showed a significant association between ageing and positivity for H. pylori. It was verified that patients aged < 10 years were 1.3 times more likely to have infection by H. pylori when compared with those aged > 10 years. Finally, no association was found between Tp53 polymorphisms and the presence of H. pylori.(AU)
Assuntos
Animais , Helicobacter pylori/imunologia , Infecções/fisiopatologia , Polimorfismo Genético , Neoplasias Gástricas , Medição de Risco/tendências , Reação em Cadeia da PolimeraseRESUMO
The risk of developing gastric cancer is believed to be related to differences among Helicobacter pylori strains and the inflammatory responses mediated by host genetic factors. H. pylori infection is acquired at an early age and in the absence of appropriate antibiotic therapy, it generally persists for life. Tp53 gene regulates the transcription of several cytokines and chemokines involved in innate immunity and its action may be influenced by the presence of different H. pylori strains. The present study aimed to detect H. pylori in pediatric patients, to access Tp53 polymorphism at codon 72 and to correlate such findings with age and histopathological results. Three hundred and forty-two patients were analyzed. DNA from their gastric biopsies was extracted and the detection of H. pylori was performed through polymerase chain reaction assays, urease test and histopathologic examination. Allelic discrimination of SNP rs1042522 (Tp53) was performed by real-time polymerase chain reaction. Our results suggest a possible relationship between the presence of H. pylori and chronic gastritis in children and young patients, and showed a significant association between ageing and positivity for H. pylori. It was verified that patients aged < 10 years were 1.3 times more likely to have infection by H. pylori when compared with those aged > 10 years. Finally, no association was found between Tp53 polymorphisms and the presence of H. pylori.
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , /genética , Helicobacter pylori , Infecções por Helicobacter/diagnóstico , Reação em Cadeia da Polimerase/métodosRESUMO
Helicobacter pylori, a gram-negative bacterium, possesses two important virulence factors: the vacuolating toxin (vacA), and the cytotoxin-associated gene product (cagA). The aim of the present study was to evaluate the presence of H. pylori in the stomach and oral cavity of humans and compare the cagA and vacA genotypes of H. pylori found in different samples (stomach, saliva and dental plaque) from the same patient. Gastric biopsies, saliva and dental plaques were obtained from 62 dyspeptic adults. DNA was extracted and evaluated for the presence of H. pylori and the alleles cagA and vacA. Persons with gastritis had a higher frequency of H. pylori -positive samples in the stomach while positive samples from gastric biopsies were significantly correlated with those from the oral cavity. There was a high H. pylori frequency in patients while the cagA gene was associated with vacA s1 alleles in gastric biopsies. Our results suggest a reservoir of the species in the oral cavity and that, in one patient, more than one H. pylori strain may exist in the saliva, dental plaque and stomach. We found a relationship between gastric infection and the bacterium in the oral cavity, with the cytotoxin genotype varying between saliva and dental plaque.(AU)
Assuntos
Humanos , Biópsia , Helicobacter pylori , Infecções por Helicobacter/diagnóstico , Saliva , Estômago , Vírus 40 dos Símios , Citotoxinas , Placa DentáriaRESUMO
This study examines the changes in cellular lipids that take place when Trypanosoma cruzi epimastigotes and metacyclic trypomastigotes are transferred from 28 to 37 degrees C. We found a rise in the sterol to phospholipid ratio, as well as in the triacylglycerol and steryl ester cellular content in T. cruzi epimastigotes. In addition, saturated to unsaturated fatty acid ratios in phospholipids increase. This latter effect appears to be due to two concurrent processes. Firstly, fatty acyl delta9 and, especially, delta12 desaturations are significantly diminished at 37 degrees C. Secondly, triacylglycerols and steryl esters undergo changes in their fatty acyl composition opposite to those simultaneously observed in phospholipids, i.e. the ratio of saturated to unsaturated fatty acids markedly decreases. Similar alterations in each of the lipid classes and in the fatty acid composition of polar and neutral lipids were found in cultured metacyclic trypomastigotes on exposure to the same shift-up. These observations suggest that a global remodeling of cellular lipids that involves extensive fatty acid exchange between neutral and polar lipid pools represents a novel and important mechanism of adaptation of the parasites to the temperature changes they encounter in their life cycle.
Assuntos
Metabolismo dos Lipídeos , Trypanosoma cruzi/metabolismo , Adaptação Fisiológica , Animais , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos/análise , Lipídeos/química , Fosfolipídeos/química , Fosfolipídeos/metabolismo , Esteróis/química , Esteróis/metabolismo , Temperatura , Triglicerídeos/química , Triglicerídeos/metabolismo , Trypanosoma cruzi/crescimento & desenvolvimentoRESUMO
We have studied the effects of lovastatin, an inhibitor of hydroxy-methylglutaryl-CoA-reductase, on cultures of Trypanosoma cruzi epimastigotes. It inhibits growth at 10 and 30 micrograms per ml; these effects are reverted by 100 microM of squalene, but not by 100 microM of cholesterol. Lovastatin at the same amounts inhibits [14C]acetate incorporatin into sterols, isolated either by digitonin precipitation or thin layer chromatography. At 50 micrograms per ml it kills most of the trypanosomes. These concentrations are below reported toxic levels for mammals; this drug and its analogs should, therefore, be tested as chemotherapeutic agents against Chagas' disease.
Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases , Lovastatina/farmacologia , Esteróis/biossíntese , Trypanosoma cruzi/crescimento & desenvolvimento , Acetatos/metabolismo , Animais , Colesterol/farmacologia , Cinética , Esqualeno/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/metabolismoRESUMO
In an attempt to extend our knowledge of the biology of feeding of the ciliateTetrahymena thermophila, this organism was grown axenically on complex organic material. The nutrient substrate was based on autoclaved wheat grains and adjusted to either pH 5.5 or 7.5. In wild type cultures the cells grew and multiplied only under acidic conditions. In cultures of a mutant cell line blocked in the secretion of acid hydrolases the cells did not grow at either pH value. Thus released acid hydrolases may play a key role in the utilization of complex nutrients in combination with uptake of small organic molecules. Mechanisms in the feeding biology ofTetrahymena thermophila andParamecium tetraurelia are compared.
RESUMO
The interaction of methylparaben and propylparaben with bilirubin-albumin complexes was studied using difference spectra, Sephadex gel filtration, red blood cell uptake of bilirubin, and the peroxidase assay. Methylparaben was found to be a weak competitor with bilirubin for binding to primary albumin-binding sites but a strong binding competitor (similar to sulfisoxazole) at secondary sites. The displacing effects of methylparaben and sulfisoxazole were additive. Propylparaben bound to albumin but did not displace bilirubin. Drugs and injectable saline and water preparations which contain methylparaben should be avoided in jaundiced newborn infants when the high-affinity albumin-binding sites approach saturation.
Assuntos
Bilirrubina/metabolismo , Parabenos/farmacologia , Albumina Sérica/metabolismo , Benzoatos/farmacologia , Sítios de Ligação , Ligação Competitiva , Composição de Medicamentos , Humanos , Recém-Nascido , Doenças do Prematuro/tratamento farmacológico , Icterícia Neonatal/tratamento farmacológico , Kernicterus/prevenção & controle , Ligação Proteica/efeitos dos fármacos , Sulfisoxazol/farmacologiaRESUMO
The effects of gentamicin on albumin binding of bilirubin and bilirubin transport were studied using Sephadex gel filtration, spectrophotometric analysis, peroxidase assay, and red blood cell uptake. No significant interaction between gentamicin and bilirubin binding or transport was observed.