RESUMO
More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)
São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)
Assuntos
Animais , Cães , Eucoccidiida/isolamento & purificação , Parasitologia , Filogenia , Epidemiologia Molecular , Reação em Cadeia da Polimerase/veterináriaRESUMO
More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)
São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)
Assuntos
Animais , Cães , Eucoccidiida/isolamento & purificação , Parasitologia , Filogenia , Epidemiologia Molecular , Reação em Cadeia da Polimerase/veterináriaRESUMO
Group A rotaviruses (RVA) are the main causing agents of acute gastroenteritis worldwide, having a great impact on childhood mortality in developing countries. The objective of this study was to identify RVA-positive fecal samples with mixed P genotypes by hemi-nested reverse transcriptase-polymerase chain reaction (RT-PCR), followed by sequencing confirmation. Our results showed that, from the 81 RVA-positive samples, 25 were positive for more than one P genotype by hemi-nested RT-PCR. Of these 25 samples, 12 (48%) had their mixed P genotypes confirmed by sequencing and, from these, 10 were identified as P[6]P[8], one as P[4]P[6], and one as P[4]P[6]P[8]. Our results confirm the occurrence of RVA mixed infections among children in Brazil and reinforce the importance of the constant monitoring of RVA circulating strains for the efficacy of control/prevention against these agents.
Assuntos
Gastroenterite/virologia , Infecções por Rotavirus/virologia , Rotavirus/classificação , Rotavirus/genética , Brasil , Criança , Fezes/virologia , Feminino , Genótipo , Humanos , Masculino , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Rotavirus/isolamento & purificação , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
Proteases perform a wide variety of functions inside and outside cells, regulating many biological processes. Infectious microorganisms use proteases, either secreted or attached to their cell surface to weaken and invade their hosts. Therefore, proteases are targets for drugs against a diverse set of diseases. Paracoccidioides brasiliensis is the most prevalent fungal pathogen causing systemic mycosis in Latin America. The development of paracoccidioidomycosis depends on interactions between fungal and host components and proteases have been described as important factors implicated in the mechanism of host colonization by fungi. The primary goal for this study is to present an overview of the transcriptome sequences--identified cDNAs that encode proteases. We obtained a total of 53 cDNAs encoding proteases; 15 were classified as ATP-independent, 12 as ATP-dependent, 22 as proteasome subunits, and 4 as deubiquitinating proteases. The mechanisms and biological activity of these proteases differ in substrate specificity and in catalytic mechanisms.
Assuntos
Humanos , DNA Complementar/análise , Paracoccidioides/enzimologia , Peptídeo Hidrolases/genética , Regulação Fúngica da Expressão Gênica/genética , Transcrição Gênica/genética , DNA Complementar/genética , Dados de Sequência Molecular , Etiquetas de Sequências Expressas , Paracoccidioides/genética , Paracoccidioides/patogenicidade , Paracoccidioidomicose/virologia , Sequência de Bases , VirulênciaRESUMO
The availability of the complete genome of the Gram-negative beta-proteobacterium Chromobacterium violaceum has increasingly impacted our understanding of this microorganism. This review focuses on the genomic organization and structural analysis of the deduced proteins of the chemosensory adaptation system of C. violaceum. C. violaceum has multiple homologues of most chemotaxis genes, organized mostly in clusters in the bacterial genome. We found at least 67 genes, distributed in 10 gene clusters, involved in the chemotaxis of C. violaceum. A close examination of the chemoreceptors methyl-accepting chemotaxis proteins (MCPs), and the deduced sequences of the members of the two-component signaling system revealed canonical motifs, described as essential for the function of the deduced proteins. The chemoreceptors found in C. violaceum include the complete repertoire of such genes described in bacteria, designated as tsr, tar, trg, and tap; 41 MCP loci were found in the C. violaceum genome. Also, the C. violaceum genome includes a large repertoire of the proteins of the chemosensory transducer system. Multiple homologues of bacterial chemotaxis genes, including CheA, CheB, CheD, CheR, CheV, CheY, CheZ, and CheW, were found in the C. violaceum genome
Assuntos
Chromobacterium/genética , Flagelos/genética , Genes Bacterianos/genética , Proteínas de Bactérias/genética , Quimiotaxia/genética , Chromobacterium/fisiologia , Flagelos/fisiologia , Genoma Bacteriano , Genes Bacterianos/fisiologia , Proteínas de Bactérias/fisiologia , Proteínas de Membrana/genética , Proteínas de Membrana/fisiologia , Quimiotaxia/fisiologiaRESUMO
Paracoccidioides brasiliensis is a pathogenic fungus that undergoes a temperature-dependent cell morphology change from mycelium (22 degrees C) to yeast (36 degrees C). It is assumed that this morphological transition correlates with the infection of the human host. Our goal was to identify genes expressed in the mycelium (M) and yeast (Y) forms by EST sequencing in order to generate a partial map of the fungus transcriptome. Individual EST sequences were clustered by the CAP3 program and annotated using Blastx similarity analysis and InterPro Scan. Three different databases, GenBank nr, COG (clusters of orthologous groups) and GO (gene ontology) were used for annotation. A total of 3,938 (Y = 1,654 and M = 2,274) ESTs were sequenced and clustered into 597 contigs and 1,563 singlets, making up a total of 2,160 genes, which possibly represent one-quarter of the complete gene repertoire in P. brasiliensis. From this total, 1,040 were successfully annotated and 894 could be classified in 18 functional COG categories as follows: cellular metabolism (44%); information storage and processing (25%); cellular processes-cell division, posttranslational modifications, among others (19%); and genes of unknown functions (12%). Computer analysis enabled us to identify some genes potentially involved in the dimorphic transition and drug resistance. Furthermore, computer subtraction analysis revealed several genes possibly expressed in stage-specific forms of P. brasiliensis. Further analysis of these genes may provide new insights into the pathology and differentiation of P. brasiliensis.
Assuntos
Etiquetas de Sequências Expressas , Genoma Fúngico , Paracoccidioides/genética , Sequência de Bases , Brasil , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Transcrição GênicaRESUMO
In Brazil, abortion is legally allowed only when it is necessary to save a woman's life or when pregnancy has occurred following rape. Despite this law, induced abortion is widely carried out. This study presents the findings as to the determinants of 2,084 abortions admitted to two major obstetric hospitals in Fortaleza, Brazil, between October 1992 and September 1993. Most of these women (2,074) have admitted an attempt to terminate pregnancy and 10 women were classified as induced abortion cases based on the findings of signs of intervention such as cervical laceration, perforation or foreign bodies in the vagina or uterus. The study findings indicate that self-administration of medicines plays an important role in terminating pregnancy. Among the 2,074 women who admitted to terminating the pregnancy 66% reported using misoprostol to induce abortion. Misoprostol, a prostaglandin E1 analogue indicated for ulcer treatment, has been widely used as an abortifacient by women in Brazil. Misoprostol has some uterine effects but it is not effective in inducing abortion. Among women who were hospitalized for complications resulting from abortion about 59.7% were 20 to 29 years old and 22.6% were aged less than 20. The majority of the women (91.6%) were Catholic and only 4.3% were illiterate. About 62% of the abortion cases lived alone or did not have a stable partner. Most of the women (59.2%) reported less than 2 live births and 11.8% had experienced a previous abortion; 61.1% of the women were not using a contraceptive method at the time of conception. The main reasons for this were "fear of side effects", "did not expect to have sexual intercourse" and "did not expect to get pregnant". The authors suggest that the situation of a high rate of self-inflicted abortion may be changed by the application of an appropriate contraceptive and reproductive health programme.