RESUMO
Gastric cancer (GC) remains one of the main causes of cancer-related death worldwide. There are two distinct histological types of GC: diffuse and intestinal. The latter is characterized by the presence of pre-neoplastic lesions. One of the most frequently altered enzymes in intestinal GC is COX-2, an important lesion marker. This work aimed to study COX-2 methylation and expression in N-methyl-N-Nitrosurea (MNU)-induced intestinal GC in six Sapajus apella animals. The partial promoter sequence of S. apella COX-2 gene was obtained and used to identify transcription factors and cis-regulatory element binding sites. The COX-2 methylation pattern was assessed using Methylation-Specific PCR (MSP), and expression was analyzed by immunohistochemistry (IHQ). A total of 20 samples were obtained. A 675 bp fragment of the S. apella COX-2 promoter region was obtained, and it was 99.2% and 68.2% similar to H. sapiens and S. boliviensis, respectively. Similar to humans, several transcription factors and cis-regulatory element binding sites were identified in the S. apella sequence. MSP revealed that all samples were methylated. However, IHQ results demonstrated positive COX-2 expression in all pre-neoplastic and tumoral samples. The results suggest that the analyzed fragment is not crucial in COX-2 regulation of GC in S. apella.
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Metopid armophoreans are ciliates commonly found in anaerobic environments worldwide; however, very little is known of their fine structure. In this study, the metopid Parametopidium circumlabens (Biggar and Wenrich 1932) Aescht, 1980, a common endocommensal of sea urchins, is investigated for the first time with emphasis on transmission electron microscopy, revealing several previously unknown elements of its morphology. Somatic dikinetids of P. circumlabens have a typical ribbon of transverse microtubules, an isolated microtubule near triplets 4 and 5 of the anterior kinetosome, plus two other microtubules between anterior and posterior kinetosomes, a short kinetodesmal striated fiber and long postciliary microtubules. In the dikinetids of the perizonal stripe, the kinetodesmal fiber is very pronounced, and there is a conspicuous microfibrillar network system associated with the kinetosomes. A new structure, shaped as a dense, roughly cylindrical mass surrounded by microtubules, is found associated with the posterior kinetosome of perizonal dikinetids. The paroral membrane is diplostichomonad and the adoral membranelles are of the "paramembranelle" type. Bayesian inference and maximum-likelihood analysis of the 18S-rDNA gene unambiguously placed P. circumlabens as sister group of the cluster formed by ((Atopospira galeata, Atopospira violacea) Metopus laminarius) + Clevelandellida, corroborating its classification within the Metopida.
Assuntos
Cilióforos/genética , Cilióforos/ultraestrutura , Ouriços-do-Mar , Simbiose , Animais , Corpos Basais/ultraestrutura , Teorema de Bayes , Cilióforos/classificação , Cilióforos/fisiologia , Microscopia Eletrônica , Microtúbulos/ultraestrutura , Filogenia , Análise de Sequência de DNARESUMO
Artibeus obscurus (Mammalia: Chiroptera) is endemic to South America, being found in at least 18 Brazilian states. Recent studies revealed that different populations of this genus present distinct phylogeographic patterns; however, very little is known on the population genetics structure of A. obscurus in the Amazon rainforest. Here, using a fragment (1010bp) of the mitochondrial gene cytochrome b from 87 samples, we investigated patterns of genetic divergence among populations of A. obscurus from different locations in the Brazilian Amazon rainforest and compared them with other Brazilian and South American regions. Analysis of molecular variance (AMOVA), fixation index (Fst) analysis, and phylogeographic patterns showed divergence between two major monophyletic groups, each one corresponding to a geographic region associated with the Atlantic and Amazon forest biomes. The Atlantic forest clusters formed a monophyletic group with a high bootstrap support and a fragmented distribution that follows the pattern predicted by the Refuge Theory. On the other hand, a different scenario was observed for the Amazon forest, where no fragmentation was identified. The AMOVA results revealed a significant geographic heterogeneity in the distribution of genetic variation, with 70% found within populations across the studied populations (Fst values ranging from 0.05864 to 0.09673; φST = 0.55). The intrapopulational analysis revealed that one population (Bragança) showed significant evidence of population expansion, with the formation of 2 distinct phylogroups, suggesting the occurrence of a subspecies or at least a different population in this region. These results also suggest considerable heterogeneity for A. obscurus in the Amazon region.
Assuntos
Quirópteros/classificação , Quirópteros/genética , Animais , Evolução Biológica , Brasil , Citocromos b/genética , Demografia , Frutas , Genes Mitocondriais , Variação Genética , Filogeografia , Análise de Sequência de DNARESUMO
Gastric cancer is a major public health problem in Pará state, where studies suggest complex genetic and epigenetic profiles of the population, indicating the need for the identification of molecular markers for this tumor type. In the present study, the methylation patterns of three genes [p16 (INK4A), p15 (INK4B), and adenomatous polyposis coli (APC)] were assessed in patients with gastric adenocarcinoma from Pará state in order to identify possible molecular markers of gastric carcinogenesis. DNA samples from tumoral and non-tumoral gastric tissues were modified with sodium bisulfite. A fragment of the promoter region of each gene was amplified and sequenced, and samples with more than 20 % of methylated CpG sites were considered hypermethylated. The correlation between the methylation pattern of the selected genes and the MTHFR C677T polymorphism, as well as the relationship between APC and CDH1 methylation, were evaluated. The results suggest that APC hypermethylation is an age-specific marker of gastric carcinogenesis, and the concordance of this event with CDH1 hypermethylation suggests that the Wnt pathway has an important role in gastric carcinogenesis. While the hypermethylation pattern of p15 (INK4B) seems to be an earlier event in this type of tumor, the hypomethylated status of this gene seems to be correlated to the C677T MTHFR TT genotype. On the other hand, the observed pattern of p16 (INK4A) hypermethylation suggests that this event is a good marker for the gastric cancer pathway in the Pará state population.
Assuntos
Proteína da Polipose Adenomatosa do Colo/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Metilação de DNA , Genes p16 , Neoplasias Gástricas/genética , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Antígenos CD , Biomarcadores Tumorais/genética , Brasil , Caderinas/genética , Ilhas de CpG , DNA de Neoplasias/genética , DNA de Neoplasias/metabolismo , Feminino , Humanos , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo de Nucleotídeo Único , Neoplasias Gástricas/metabolismo , Via de Sinalização WntRESUMO
Morphology of the urostylid ciliate Hemicycliostyla sphagni Stokes, 1886, the type of Hemicycliostyla Stokes, 1886, is investigated based on live and protargol-impregnated specimens from a Brazilian population. The absence of transverse cirri, which has been considered the main diagnostic feature of Hemicycliostyla, separating it from Pseudourostyla Borror, 1972, was found to vary within the studied population, with 50% of the specimens exhibiting inconspicuous and/or rudimentary transverse cirri. A redefinition of Hemicycliostyla was possible based on combined features of interphase and divisional morphogenesis: Retroextendia Berger, 2006, with bi- or multicoronal frontal cirral pattern; fronto-terminal cirri present; multiple left and right marginal cirral rows that replicate independently via within-row development, each parental row producing one primordium per divider; caudal cirri lacking; and presence/absence of transverse cirri may be intrapopulationally variable. Phylogenetic analyses of the 18S rDNA marker unambiguously placed H. sphagni as sister group of Pseudourostyla franzi Foissner, 1987, which is herein transferred to Hemicycliostyla as Hemicycliostyla franzi comb. nov.
Assuntos
Cilióforos/citologia , Cilióforos/genética , Filogenia , Brasil , Cilióforos/classificação , Análise por Conglomerados , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Genes de RNAr , Dados de Sequência Molecular , RNA de Protozoário/genética , RNA Ribossômico/genética , Análise de Sequência de DNARESUMO
UNLABELLED: The aim of this study was to investigate the protein expression and methylation pattern of P21(CIP1) and P27(KIP1) genes. PATIENTS AND METHODS: Twenty samples of gastric tumor and non-tumoral tissues of patients from Pará state, Brazil were collected. Methylation patterns were assessed by bisulfite sequencing, and protein expression evaluated with immunohistochemical analysis. RESULTS: None of the analyzed samples showed methylation in both genes. Immunohistochemistry analysis demonstrated mislocalization or absence of expression of P21(CIP1) and P27(KIP1) in 7/20 and 6/20 of the studied samples, respectively. No correlations regarding protein expression and clinicopathological characteristics were observed; down-regulation of expression of P21(CIP1) with low (I-II) tumor stage (p=0.0777), and older age (>50 years old) with negative or mislocalization of P27(KIP1) (p=0.0922) were of borderline the statistical significance. CONCLUSION: Our results suggest that hypermethylation does not contribute to P21(CIP1) and P27(KIP1) silencing in gastric cancer, and that the role of these genes in the gastric tumorigenesis pathways should be studied further in the Pará state population.
Assuntos
Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Neoplasias Gástricas/patologia , Adulto , Idoso , Brasil , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Metilação de DNA , DNA de Neoplasias/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Metilação , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismoRESUMO
BACKGROUND/AIM: In female dogs, mammary cancer is the most frequent cancer type, accounting for 50% of all tumors affecting these animals. Amongst the commonly altered genes in cancer is the cell-cycle regulator cyclin-dependent kinase inhibitor 2B (Cdkn2b), whose expression is negatively regulated by protein products of BMI1 proto-oncogene (Bmi1), MYC proto-oncogene (Myc) and T-box gene transcription factor 2 (Tbx2) genes. Considering this, the aim of this study was to evaluate the expression pattern of the Cdkn2b gene and these regulators in canine mammary tumors of dogs from Northern Brazil (Belém, Pará). MATERIAL AND METHODS: Gene expression in samples from 33 animals was assessed by real-time polymerase chain reaction. To check the influence of methylation on gene expression, bisulfite sequencing polymerase chain reaction was also performed. RESULTS: All studied genes, except Cdkn2b, were found at increased expression levels in tumor tissue when compared with control samples. No correlation between expression and methylation data was observed. CONCLUSION: Our results suggest these markers may have a diagnostic value in the veterinary clinic.
Assuntos
Inibidor de Quinase Dependente de Ciclina p15/genética , Doenças do Cão/genética , Neoplasias Mamárias Animais/genética , Proteína Quinase 7 Ativada por Mitógeno/genética , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas com Domínio T/genética , Animais , Metilação de DNA , Cães , Epigênese Genética , Feminino , Perfilação da Expressão Gênica/veterinária , Regulação Neoplásica da Expressão Gênica , Proto-Oncogene Mas , Análise de Sequência de DNA/veterináriaRESUMO
Glycosyltransferase 6 gene family includes ABO, Ggta1, iGb3S, and GBGT1 genes and by three putative genes restricted to mammals, GT6m6, GTm6, and GT6m7, only the latter is found in primates. GT6 genes may encode functional and nonfunctional proteins. Ggta1 and GBGT1 genes, for instance, are pseudogenes in catarrhine primates, while iGb3S gene is only inactive in human, bonobo, and chimpanzee. Even inactivated, these genes tend to be conversed in primates. As some of the GT6 genes are related to the susceptibility or resistance to parasites, we investigated (i) the selective pressure on the GT6 paralogs genes in primates; (ii) the basis of the conservation of iGb3S in human, chimpanzee, and bonobo; and (iii) the functional potential of the GBGT1 and GT6m7 in catarrhines. We observed that the purifying selection is prevalent and these genes have a low diversity, though ABO and Ggta1 genes have some sites under positive selection. GT6m7, a putative gene associated with aggressive periodontitis, may have regulatory function, but experimental studies are needed to assess its function. The evolutionary conservation of iGb3S in humans, chimpanzee, and bonobo seems to be the result of proximity to genes with important biological functions.
RESUMO
Glioblastomas, also known as glioblastoma multiforme (GBM), are the most aggressive and malignant type of primary brain tumor in adults, exhibiting notable variability at the histopathological, genetic and epigenetic levels. Recently, epigenetic alterations have emerged as a common hallmark of many tumors, including GBM. Considering that a deeper understanding of the epigenetic modifications that occur in GBM may increase the knowledge regarding the tumorigenesis, progression and recurrence of this disease, in this review we discuss the recent major advances in GBM epigenetics research involving histone modification, glioblastoma stem cells, DNA methylation, noncoding RNAs expression, including their main alterations and the use of epigenetic therapy as a valid option for GBM treatment.
Assuntos
Neoplasias Encefálicas/genética , Epigênese Genética , Glioblastoma/genética , Biomarcadores Tumorais/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Regulação Neoplásica da Expressão Gênica , Glioblastoma/metabolismo , Glioblastoma/patologia , HumanosRESUMO
Astrocytic gliomas, which are derived from glial cells, are considered the most common primary neoplasias of the central nervous system (CNS) and are histologically classified as low grade (I and II) or high grade (III and IV). Recent studies have shown that astrocytoma formation is the result of the deregulation of several pathways, including the RB/E2F pathway, which is commonly deregulated in various human cancers via genetic or epigenetic mechanisms. On the basis of the assumption that the study of the mechanisms controlling the INK4/ARF locus can help elucidate the molecular pathogenesis of astrocytic tumors, identify diagnostic and prognostic markers, and help select appropriate clinical treatments, the present study aimed to evaluate and compare methylation patterns using bisulfite sequencing PCR and evaluate the gene expression profile using real-time PCR in the genes CDKN2A, CDKN2B, CDC6, Bmi-1, CCND1, and RB1 in astrocytic tumors. Our results indicate that all the evaluated genes are not methylated independent of the tumor grade. However, the real-time PCR results indicate that these genes undergo progressive deregulation as a function of the tumor grade. In addition, the genes CDKN2A, CDKN2B, and RB1 were underexpressed, whereas CDC6, Bmi-1, and CCND1 were overexpressed; the increase in gene expression was significantly associated with decreased patient survival. Therefore, we propose that the evaluation of the expression levels of the genes involved in the RB/E2F pathway can be used in the monitoring of patients with astrocytomas in clinical practice and for the prognostic indication of disease progression.
Assuntos
Astrocitoma/genética , Neoplasias Encefálicas/genética , Fatores de Transcrição E2F/genética , Regulação Neoplásica da Expressão Gênica/genética , Proteína do Retinoblastoma/genética , Transdução de Sinais/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Astrocitoma/patologia , Neoplasias Encefálicas/patologia , Proteínas de Ciclo Celular/genética , Ciclina D1/genética , Inibidor de Quinase Dependente de Ciclina p15/genética , Inibidor p16 de Quinase Dependente de Ciclina/genética , Metilação de DNA/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/genética , Complexo Repressor Polycomb 1/genética , Prognóstico , Adulto JovemRESUMO
Pseudokeronopsidae Borror & Wicklow, 1983 are biotechnologically important ciliate protists which produce toxic defense substances; however, their diversity is still little known in Brazil. In the present study, Tetrakeronopsis silvanetoi, a new genus and species of marine pseudokeronopsid hypotrichs is described from samples of water with bottom sediment collected from the coast of São Paulo state. Its phylogenetic affinities to the "core urostyloids" are hypothesized based on analyses of the 18S-rDNA marker, and a new subfamily, the Nothoholostichinae subfam. nov., is erected to name the monophylum composed of pseudokeronopsids in which the anterior corona is usually formed by four frontal cirri. In addition, the new combination Monocoronella longissima comb. nov. is proposed for Nothoholosticha longissima (Dragesco & Dragesco-Kernéis, 1986) Li et al., 2009.
Assuntos
Cilióforos/genética , Cilióforos/isolamento & purificação , Brasil , DNA de Protozoário/genética , DNA Ribossômico/genética , FilogeniaRESUMO
Ciliates occur in all major aquatic and soil environments worldwide and are important links in the microbial food webs, which, along with other free-living protists, are generally overlooked in biodiversity conservation programs. In Brazil, the northern region comprises the Brazilian Amazonia, an area widely known for its huge biodiversity. However, the diversity of ciliates in that region is still almost unknown. As result of the present study, a total of 21 species of ciliates, distributed among 15 genera, were inventoried from samples of eutrophized water collected in the city of Belém, capital of the state of Pará, one of the states which comprise the Brazilian Amazonia. In addition, a local population of the rare scuticociliate Cristigera hammeri is described from optical and electron microscopy observations, and its synonymy with C. pleuronemoides is rejected based on new evidence.
Assuntos
Biodiversidade , Cilióforos/classificação , Cilióforos/citologia , Água/parasitologia , Brasil , Cilióforos/ultraestrutura , Eutrofização , Oligoimenóforos/classificação , Oligoimenóforos/citologia , Oligoimenóforos/ultraestrutura , Especificidade da EspécieRESUMO
Gastric cancer, despite its decline in incidence, remains a public health problem worldwide, especially in Brazil, where higher incidence indexes are still described. The Survivin gene codifies a multifunctional protein involved in the regulation of the cell cycle and inhibition of the apoptotic pathway, and a polymorphism (-31C/G) located in its promoter region is associated with gene regulation. In order to evaluate the correlation of this polymorphism with gastric cancer risk in a northern Brazil population, we sequenced a fragment containing the polymorphism in individuals with gastric cancer and controls. We observed no differences of alleles and genotype frequencies between cases and controls. However, G carriers of the tumor group had an increased relative risk of developing tumors of diffuse type (OR: 2.22-IC 95%: 0.4835-10.2137), localized in the antrum (OR: 2.16-IC 95%: 0.4811-9.6971) and in younger patients (<50 years-old) (OR: 3.65-IC 95%: 0.4012-33.2429), although with no statistical significance. Nevertheless, C carriers with a high D17S250 microsatellite instability (TP53 gene) show a higher risk to develop gastric tumors (P = 0.0453; OR: 4.1556-IC95%: 0.9716-17.7728), suggesting that the mutate TP53 gene may fail in control and inhibition of Survivin expression, favoring the gastric carcinogenesis. The present result suggests that the presence of the C allele of -31C/G Survivin promoter polymorphism in combination with D17S250 instability may be used as a risk factor for gastric cancer in our population. However, other studies based on a larger sample size are required to properly assess such hypothesis.
Assuntos
Proteínas Inibidoras de Apoptose/genética , Polimorfismo de Nucleotídeo Único , Neoplasias Gástricas/genética , Proteína Supressora de Tumor p53/genética , Adulto , Idoso , Brasil , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Instabilidade de Microssatélites , Pessoa de Meia-Idade , Regiões Promotoras Genéticas , Risco , Neoplasias Gástricas/patologia , SurvivinaRESUMO
The ciliate Parasterkiella thompsoni (Foissner, 1996) nov. gen., nov. comb. was originally described from Antarctica. In the present study, we report the morphology, morphogenesis during cell division, and molecular phylogeny inferred from the 18S-rDNA sequence of a population isolated from the Rancho Hambre peat bog, Tierra del Fuego Province (Argentina). The study is based on live and protargol-impregnated specimens. Molecular phylogeny was inferred from trees constructed by means of the maximum parsimony, neighbor joining, and Bayesian analyses. The interphase morphology matches the original description of the species. During the cell division, stomatogenesis begins with the de novo proliferation of two fields of basal bodies, each one left of the postoral ventral cirri and of transverse cirri, which later unify. Primordia IV-VI of the proter develop from disaggregation of cirrus IV/3, while primordium IV of the opisthe develops from cirrus IV/2 and primordia V and VI from cirrus V/4. Dorsal morphogenesis occurs in the Urosomoida pattern-that is, the fragmentation of kinety 3 is lacking. Three macronuclear nodules are generated before cytokinesis. Phylogenetic analyses consistently placed P. thompsoni within the stylonychines. New data on the morphogenesis of the dorsal ciliature justifies the transference of Sterkiella thompsoni to a new genus Parasterkiella.
Assuntos
Cilióforos/classificação , Cilióforos/isolamento & purificação , Filogenia , Microbiologia do Solo , Argentina , Cilióforos/citologia , Cilióforos/genética , Análise por Conglomerados , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Genes de RNAr , Microscopia , Dados de Sequência Molecular , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Análise de Sequência de DNARESUMO
UNLABELLED: The aim of this study was to verify genetic and epigenetic alterations in gastric cancer patients from Pará state, northern Brazil. MATERIALS AND METHODS: Exon 11 of KIT and two promoter polymorphisms (-160 C/A and -347 G/GA) of the E-cadherin gene (CDH1), and their correlation with the promoter methylation status were analyzed. RESULTS: No genetic alterations in KIT were found. Promoter polymorphisms revealed an increased probability of developing gastric cancer, especially of the diffuse-type, in patients carrying -160 A and -347 GA alleles. Analyses of CDH1 methylation suggested a significant difference between hypermethylated and non-hypermethylated samples, with a positive association between the -160 A allele and hypermethylation. CONCLUSION: Our results suggest that -160 A and -347 GA polymorphisms may increase the chance of developing gastric cancer in the studied population and that -160 A polymorphism seems to be related to the hypermethylation pattern of the promoter region of CDH1.
Assuntos
Caderinas/genética , Metilação de DNA , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas c-kit/genética , Neoplasias Gástricas/genética , Adulto , Idoso , Ilhas de CpG , Feminino , Frequência do Gene , Haplótipos , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Piper tuberculatum is an exotic Piper from the Amazon region that shows resistance to infection by Fusarium solani f. sp. piperis, causal agent of Fusarium disease in black pepper (Piper nigrum L.). In this work we aimed to study the interaction between P. tuberculatum and F. solani f. sp. piperis at a molecular level, using suppression subtractive hybridization to identify genes potentially related to Fusarium disease resistance. Comparative sequence analysis confirmed that clones isolated here show a high identity with genes coding for proteins that have a known role in plant defense response mechanisms, such as peroxidase, hydroxyproline-rich glycoprotein and CBL-interacting protein kinase. The present study constitutes the first effort to understand the molecular basis of this plant-pathogen interaction, identifying genes which may be used in the future genetic improvement of black pepper.
Assuntos
Fusarium , Hibridização de Ácido Nucleico/métodos , Piper/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Sequência de Bases , Glicoproteínas/metabolismo , Peroxidase/metabolismo , Piper/enzimologia , Proteínas Quinases/metabolismo , Retroelementos/genética , Análise de Sequência de DNARESUMO
The present study verified the quality of the water of Água Preta Lake, which is part of the public supply for the Northern Brazilian municipality of Belém (Pará State). Six samples were collected from each of six sampling points. The concentration (NMP) of coliforms was determined by Multiple-tube fermentation Technique. The isolates of Escherichia coli obtained from the samples were tested for susceptibility to the antibiotics (cefoxitin, ampicillin, imipenem, gentamicin, and amikacin). Furthermore, was analyzed the presence of diagnostic genes for the diarrheagenic strains of E. coli. None of the genes was identified, however, the recorded concentrations of thermo-tolerant coliforms were within the recommended limits for standing water sources used for public supplies. Nevertheless, the highest concentrations of total and thermo-tolerant coliforms were recorded at two points, one adjacent to the most densely-populated area of the lake margin, and the other near the catchment area for water from the Guamá River. The susceptibility testing indicated the presence of six resistance phenotype profiles, including multi-resistant strains. The results of the study reinforce the need for the systematic monitoring of this water source, in order to provide guidelines for the development of effective management policies for public water supplies, as well as the prevention of water-borne diseases.
Neste estudo, a qualidade da água foi verificada no manancial de abastecimento Água Preta, do município de Belém (PA). Houve seis amostragens em seis pontos de coleta e a concentração de coliformes foi verificada através da Técnica de Fermentação em Tubos Múltiplos para a determinação do NMP. Os isolados de Escherichia coli obtidos foram submetidos ao teste de sensibilidade aos seguintes antimicrobianos: cefoxitina, ampicilina, imipenem, gentamicina e amicacina. Além disso, foi investigado genes codificadores de fatores de virulência relacionados às variedades diarreiogênicas de E. coli. Não houve ocorrência de genes relacionados à patogenicidade, e as concentrações de coliformes termotolerantes apresentaram-se dentro dos padrões para mananciais de superfície usados para fins de abastecimento público. Contudo, as maiores concentrações de coliformes totais e termotolerantes foram observadas no ponto de coleta próximo à captação no rio Guamá e na área de maior adensamento populacional no entorno do lago. O teste de suscetibilidade dos isolados E. coli indicou uma alta porcentagem de resistência a ampicilina, a presença de seis perfis fenotípicos e a ocorrência de multiresistência. Assim, os resultados reforçam a necessidade do monitoramento sistemático deste manancial, visando a implementação de políticas de preservação e proteção dos mananciais utilizados para fins de abastecimento público, assim como a prevenção de doenças veiculadas pela água.
RESUMO
Gastric cancer is the forth malignancy in frequency in the world. In the northern Brazil is the second neoplasia most frequent in males and the third most frequent in females. Genetic and epigenetic alterations are evolved on gastric carcinogenesis and DNA methylation is the epigenetic alteration better studied. We analyzed de novo DNA methyltransferases methylation pattern and its association with RUNX3 gene methylation pattern in Brazilian samples of intestinal-type and diffuse-type of gastric cancer. PCR methylation specific was used to evaluate DNA methylation pattern. Sixty-six samples were studied in this work. Only the gene RUNX3 presented altered methylation pattern, being methylated in 38.5% of gastric cancer intestinal-type samples and in 70% of gastric cancer diffuse-type samples and, by this reason, it should be evolved in the genesis of this neoplasia. There was a statistically significant difference among diffuse-type and intestinal-type samples (p=0.0418) and among normal and tumour tissues (p<0.0001) for RUNX3 gene but not to DNMT3A, DNMT3B e DNMT3 genes on CpG islands analyzed. Alteration of RUNX3 methylation pattern is not associated to de novo alteration of DNA methyltransferases methylation pattern on studied regionsTherefore, it becomes necessary a better comprehension of this phenomenon on gastric carcinogenesis.
El cáncer gástrico es la cuarta patología más frecuente en el mundo. En el norte del Brasil, es la segunda neoplasia más frecuente en hombres y la tercera en mujeres. Alteraciones genéticas y epigenéticas relacionadas con la carcinogénesis gástrica y la metilación del DNA son las alteraciones epigenéticas mejor estudiadas. En este trabajo, analizamos el estado de novo de metilación de genes DNA metiltransferases y su asociación con el estado de metilación del gen RUNX3 en muestras de individuos brasileños con cáncer gástrico de los tipos intestinal y difuso. Fue usada la Reacción en Cadena de la Polimerasa (PCR), metilación específica, para analizar el estado de metilación del DNA. Fueron estudiados 66 tejidos tumorales. Solamente el gen RUNX3 presentó un estado de metilación alterado, estuvo metilado en 38,5% de las muestras de cáncer gástrico tipo intestinal y en 70% de muestras de cáncer gástrico tipo difuso, lo que sugiere que estaría relacionado con la génesis de esta neoplasia. Hubo una diferencia estadística significativa entre muestras de los tipos difuso e intestinal (p=0.0418) y entre tejidos normal y tumoral (p<0.0001)parael gen RUNX3. Esta asociación no fue encontrada para los genes DNMT3A, DNMT3B y DNMT3 en las islas CpG analizadas. Alteraciones del estado de metilación de RUNX3 no están asociadas con alteraciones de novo de genes DNA metiltransferases. De esta forma se hace necesaria una mejor comprensión de este fenómeno en la carcinogénesis gástrica.
Assuntos
Humanos , Neoplasias Gástricas/genética , Adenocarcinoma/genética , Subunidade alfa 3 de Fator de Ligação ao Core/genética , Metiltransferases/genética , Reação em Cadeia da Polimerase/métodos , Metilação de DNARESUMO
In the present study, the coding region of the H gene was sequenced and analyzed in fourteen genera of New World primates (Alouatta, Aotus, Ateles, Brachyteles, Cacajao, Callicebus, Callithrix, Cebus, Chiropotes, Lagothrix, Leontopithecus, Pithecia, Saguinus, and Saimiri), in order to investigate the evolution of the gene. The analyses revealed that this coding region contains 1,101 nucleotides, with the exception of Brachyteles, the callitrichines (Callithrix, Leontopithecus, and Saguinus) and one species of Callicebus (moloch), in which one codon was deleted. In the primates studied, the high GC content (63 percent), the nonrandom distribution of codons and the low evolution rate of the gene (0.513 substitutions/site/MA in the order Primates) suggest the action of a purifying type of selective pressure, confirmed by the Z-test. Our analyses did not identify mutations equivalent to those responsible for the H-deficient phenotypes found in humans, nor any other alteration that might explain the lack of expression of the gene in the erythrocytes of Neotropical monkeys. The phylogenetic trees obtained for the H gene and the distance matrix data suggest the occurrence of divergent evolution in the primates.
Assuntos
Evolução Molecular , Fucosiltransferases , PrimatasRESUMO
Amostras de sangue e saliva foram coletadas de 76 animais da espécie Saimiri boliviensis boliviensis da Bolivia. As amostras de saliva foram testadas para a presença de antígenos ABH humanos, pelo método convencional de inibiçäo da hemaglutinaçäo. Cinquenta e nove animais foram classificados como sendo do grupo sanguíneo A e 17 como sendo do grupo AB. A distribuiçäo fenotípica ajusta-se ao esperado, assumindo-se o equilíbrio de hardy-Weinberg, em um modelo com dois alelos, com as seguintes freqüências: I A= 0.89 e I B = 0.11. Esses resultados näo mostraram diferenças estatisticamente significantes, quando comparados com aqueles descritos por Schneider et al. (unpublished data) em S. b. peruviensis. Amostras de soro de 71 animais foram utilizadas na investigaçäo de aglutininas naturais, pelo teste de hemaglutinaçäo direta e foi observado 34% de discordância entre os fenótipos salivar e sérico