RESUMEN
Proinflammatory cytokines are critical mediators that control Mycobacterium tuberculosis (Mtb) growth during active tuberculosis (ATB). To further inhibit bacterial proliferation in diseased individuals, drug inhibitors of cell wall synthesis such as isoniazid (INH) are employed. However, whether INH presents an indirect effect on bacterial growth by regulating host cytokines during ATB is not well known. To examine this hypothesis, we used an in vitro human granuloma system generated with primary leukocytes from healthy donors adapted to model ATB. Intense Mtb proliferation in cell cultures was associated with monocyte/macrophage activation and secretion of IL-1ß and TNF. Treatment with INH significantly reduced Mtb survival, but altered neither T-cell-mediated Mtb killing, nor production of IL-1ß and TNF. However, blockade of both IL-1R1 and TNF signaling rescued INH-induced killing, suggesting synergistic roles of these cytokines in mediating control of Mtb proliferation. Additionally, mycobacterial killing by INH was highly dependent upon drug activation by the pathogen catalase-peroxidase KatG and involved a host PI3K-dependent pathway. Finally, experiments using coinfected (KatG-mutated and H37Rv strains) cells suggested that active INH does not directly enhance host-mediated killing of Mtb. Our results thus indicate that Mtb-stimulated host IL-1 and TNF have potential roles in TB chemotherapy.
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Antituberculosos/farmacología , Interleucina-1beta/inmunología , Isoniazida/farmacología , Mycobacterium tuberculosis/efectos de los fármacos , Receptores de Interleucina-1/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Proteínas Bacterianas/metabolismo , Células Cultivadas , Humanos , Macrófagos/metabolismo , Monocitos/metabolismo , Fosfatidilinositol 3-Quinasas/inmunología , Receptores de Interleucina-1/antagonistas & inhibidores , Tuberculosis/tratamiento farmacológico , Tuberculosis/inmunología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidoresRESUMEN
Epithelial cell invasion by the protozoan parasite Trypanosoma cruzi is enhanced by the presence of an enzyme expressed on its cell surface during the trypomastigote life cycle stage. The enzyme, trans-sialidase (TS), is a member of one of the largest gene families expressed by the parasite and the role of its activity in mediating epithelial cell entry has not hitherto been understood. Here we show that the T. cruzi TS generates an eat me signal which is capable of enabling epithelial cell entry. We have utilized purified, recombinant, active (TcTS) and inactive (TcTS2V0) TS coated onto beads to challenge an epithelial cell line. We find that TS activity acts upon G protein coupled receptors present at the epithelial cell synapse with the coated bead, thereby enhancing cell entry. By so doing, we provide evidence that TS proteins bind glycans, mediate the formation of distinct synaptic domains and promote macropinocytotic uptake of microparticles into a perinuclear compartment in a manner which may emulate entosis.
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Endocitosis , Células Epiteliales/metabolismo , Glicoproteínas/metabolismo , Neuraminidasa/metabolismo , Animales , Membrana Celular/metabolismo , Perros , Entosis , Células Epiteliales/enzimología , Células Epiteliales/fisiología , Células de Riñón Canino Madin Darby , Microesferas , Polisacáridos/metabolismo , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.
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Genoma de Planta , Herbaspirillum/genética , Cromosomas de las Plantas , Herbaspirillum/metabolismo , Interacciones Huésped-Patógeno , Fijación del Nitrógeno , Presión Osmótica , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Protozoa and fungi are known to have extraordinarily diverse mechanisms of genetic exchange. However, the presence and epidemiological relevance of genetic exchange in Trypanosoma cruzi, the agent of Chagas disease, has been controversial and debated for many years. Field studies have identified both predominantly clonal and sexually recombining natural populations. Two of six natural T. cruzi lineages (TcV and TcVI) show hybrid mosaicism, using analysis of single-gene locus markers. The formation of hybrid strains in vitro has been achieved and this provides a framework to study the mechanisms and adaptive significance of genetic exchange. Using whole genome sequencing of a set of experimental hybrids strains, we have confirmed that hybrid formation initially results in tetraploid parasites. The hybrid progeny showed novel mutations that were not attributable to either (diploid) parent showing an increase in amino acid changes. In long-term culture, up to 800 generations, there was a variable but gradual erosion of progeny genomes towards triploidy, yet retention of elevated copy number was observed at several core housekeeping loci. Our findings indicate hybrid formation by fusion of diploid T. cruzi, followed by sporadic genome erosion, but with substantial potential for adaptive evolution, as has been described as a genetic feature of other organisms, such as some fungi.
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Enfermedad de Chagas , Trypanosoma cruzi , Enfermedad de Chagas/parasitología , ADN Protozoario/genética , Variación Genética , Genotipo , Humanos , Hibridación Genética , Hibridación de Ácido Nucleico , Trypanosoma cruzi/genéticaRESUMEN
Despite the economic impact of the infectious myonecrosis virus (IMNV) on shrimp farms in several countries, no method for immunological detection is currently available. With the aim of developing immunodiagnostic methods for IMNV detection in infected shrimps, a recombinant fragment of the IMNV major capsid protein gene encoding amino acids 105-297 (rIMNV105â297 was heterologously expressed in Escherichia coli and used to immunize Balb/c mice, generating monoclonal antibodies (MAbs). Six hybridomas were obtained, and four of these recognized the presence of IMNV in tissue homogenates from naturally infected shrimps by immunodot blot assay. Among these MAbs, three were able to detect a ~100-kDa protein, which corresponds to the predicted mass of the IMNV major capsid protein, as well as viral inclusion bodies in muscle fibroses by western blot and immunohistochemistry. Two MAbs showed high specificity and sensitivity, showing no cross-reaction with healthy shrimp tissues in any assays, indicating their usefulness for IMNV detection.
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Anticuerpos Monoclonales/inmunología , Proteínas de la Cápside/inmunología , Inmunoensayo/veterinaria , Penaeidae/virología , Totiviridae/aislamiento & purificación , Animales , Clonación Molecular , Inmunohistoquímica , Integumento Común/virología , Ratones , Ratones Endogámicos BALB C , ARN ViralRESUMEN
Trypanosoma cruzi, a zoonotic kinetoplastid protozoan parasite, is the causative agent of American trypanosomiasis (Chagas disease). Having a very plastic, repetitive and complex genome, the parasite displays a highly diverse repertoire of surface molecules, with pivotal roles in cell invasion, immune evasion and pathogenesis. Before 2016, the complexity of the genomic regions containing these genes impaired the assembly of a genome at chromosomal level, making it impossible to study the structure and function of the several thousand repetitive genes encoding the surface molecules of the parasite. We here describe the genome assembly of the Sylvio X10/1 genome sequence, which since 2016 has been used as a reference genome sequence for T. cruzi clade I (TcI), produced using high coverage PacBio single-molecule sequencing. It was used to analyze deep Illumina sequence data from 34 T. cruzi TcI isolates and clones from different geographic locations, sample sources and clinical outcomes. Resolution of the surface molecule gene distribution showed the unusual duality in the organization of the parasite genome, a synteny of the core genomic region with related protozoa flanked by unique and highly plastic multigene family clusters encoding surface antigens. The presence of abundant interspersed retrotransposons in these multigene family clusters suggests that these elements are involved in a recombination mechanism for the generation of antigenic variation and evasion of the host immune response on these TcI strains. The comparative genomic analysis of the cohort of TcI strains revealed multiple cases of such recombination events involving surface molecule genes and has provided new insights into T. cruzi population structure.
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Variación Antigénica , Trypanosoma cruzi , Familia de Multigenes , Sintenía , Trypanosoma cruzi/genéticaRESUMEN
ProtozoaDB (http://www.biowebdb.org/protozoadb) is being developed to initially host both genomics and post-genomics data from Plasmodium falciparum, Entamoeba histolytica, Trypanosoma brucei, T. cruzi and Leishmania major, but will hopefully host other protozoan species as more genomes are sequenced. It is based on the Genomics Unified Schema and offers a modern Web-based interface for user-friendly data visualization and exploration. This database is not intended to duplicate other similar efforts such as GeneDB, PlasmoDB, TcruziDB or even TDRtargets, but to be complementary by providing further analyses with emphasis on distant similarities (HMM-based) and phylogeny-based annotations including orthology analysis. ProtozoaDB will be progressively linked to the above-mentioned databases, focusing in performing a multi-source dynamic combination of information through advanced interoperable Web tools such as Web services. Also, to provide Web services will allow third-party software to retrieve and use data from ProtozoaDB in automated pipelines (workflows) or other interoperable Web technologies, promoting better information reuse and integration. We also expect ProtozoaDB to catalyze the development of local and regional bioinformatics capabilities (research and training), and therefore promote/enhance scientific advancement in developing countries.
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Bases de Datos Genéticas , Genoma de Protozoos , Animales , Gráficos por Computador , Entamoeba histolytica/genética , Genómica , Internet , Leishmania major/genética , Plasmodium falciparum/genética , Proteínas Protozoarias/química , Programas Informáticos , Trypanosoma brucei brucei/genética , Trypanosoma cruzi/genética , Interfaz Usuario-ComputadorRESUMEN
Plasmid-mediated polymyxin resistance has become a global health concern, not only because its dissemination has occurred drastically but also because it has begun to be reported in multidrug-resistant (MDR) pathogens. We hereby report microbiological and genomic characteristics of two mcr-1.1-positive polymyxin-resistant Escherichia coli isolates identified for the first time in community patients, in Santa Catarina, Southern Brazil. E. coli strains belonging to ST206 and ST354 and the resistome analysis revealed the presence of clinically important genes responsible for MDR profile. Interestingly, in both polymyxin-resistant E. coli strains, mcr-1.1 genes were carried by IncX4 plasmids, responsible for the worldwide dissemination of mcr-type genes. In this regard, plasmid backbones were almost identical to the first IncX4 plasmid reported in Brazil and sharing more than 99.9% identity to IncX4 plasmids from China, also lacking the ISApl1 insertion sequence upstream of mcr-1. In conclusion, these data confirm the presence of international ST206 and ST354 carrying mcr-1.1 genes and that the IncX4 plasmids have been key vectors contributing to the endemic status of mcr-1.1-positive polymyxin-resistant E. coli in Brazil. Also, we described the first known clinical isolate with the mrc1.1 gene in Santa Catarina state, Brazil, showing that plasmid-mediated polymyxin resistance has been affecting humans earlier than has been known so far.
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Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Plásmidos/genética , Polimixinas/farmacología , Brasil , Escherichia coli/efectos de los fármacos , Genómica/métodos , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Pacientes AmbulatoriosRESUMEN
BACKGROUND: Anopheles (Kerteszia) cruzii was the most important vector of human malaria in southern Brazil between 1930-1960. Nowadays it is still considered an important Plasmodium spp. vector in southern and south-eastern Brazil, incriminated for oligosymptomatic malaria. Previous studies based on the analysis of X chromosome banding patterns and inversion frequencies in An. cruzii populations from these areas have suggested the occurrence of three sibling species. In contrast, two genetically distinct groups among An. cruzii populations from south/south-east and north-east Brazil have been revealed by isoenzyme analysis. Therefore, An. cruzii remains unclear. METHODS: In this study, a partial sequence of the timeless gene (approximately 400 bp), a locus involved in the control of circadian rhythms, was used as a molecular marker to assess the genetic differentiation between An. cruzii populations from six geographically distinct areas of Brazil. RESULTS: The timeless gene revealed that An. cruzii from Itaparica Island, Bahia State (north-east Brazil), constitutes a highly differentiated group compared with the other five populations from south and south-east Brazil. In addition, significant genetic differences were also observed among some of the latter populations. CONCLUSION: Analysis of the genetic differentiation in the timeless gene among An. cruzii populations from different areas of Brazil indicated that this malaria vector is a complex of at least two cryptic species. The data also suggest that further work might support the occurrence of other siblings within this complex in Brazil.
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Anopheles/clasificación , Anopheles/genética , Variación Genética/genética , Insectos Vectores/genética , Malaria/transmisión , Animales , Anopheles/parasitología , Brasil , Femenino , Humanos , Insectos Vectores/clasificación , Malaria/epidemiología , Datos de Secuencia Molecular , Plasmodium/genética , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Triatoma brasiliensis is the most important autochthon vector of Trypanosoma cruzi in Brazil, where it is widely distributed in the semiarid areas of the Northeast. In order to advance the knowledge of the salivary biomolecules of Triatominae, a salivary gland cDNA library of T. brasiliensis was mass sequenced and analyzed. Polypeptides were sequenced by HPLC/Edman degradation experiments. Then 1712 cDNA sequences were obtained and grouped in 786 clusters. The housekeeping category had 24.4% and 17.8% of the clusters and sequences, respectively. The putatively secreted category contained 47.1% of the clusters and 68.2% of the sequences. Finally, 28.5% of the clusters, containing 14% of all sequences, were classified as unknown. The sialoma of T. brasiliensis showed a high amount and great variety of different lipocalins (93.8% of secreted proteins). Remarkably, a great number of serine proteases that were not observed in previous blood-sucking sialotranscriptomes were found. Nine Kazal peptides were identified, among them one with high homology to the tabanid vasodilator vasotab, suggesting that the Triatoma vasodilator could be a Kazal protein.
Asunto(s)
Biología Computacional , Biblioteca de Genes , Proteínas de Insectos/genética , Saliva/química , Glándulas Salivales/química , Triatoma/metabolismo , Secuencia de Aminoácidos , Animales , Apirasa/química , Sangre , Brasil , Clonación Molecular , Ingestión de Alimentos , Hemoproteínas , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Insectos Vectores , Filogenia , Glándulas Salivales/fisiología , Proteínas y Péptidos Salivales , Alineación de Secuencia , Análisis de Secuencia de ADN , Serina Endopeptidasas/aislamiento & purificación , Serina Endopeptidasas/metabolismo , Triatoma/clasificación , Triatoma/genética , Trypanosoma cruzi/fisiologíaRESUMEN
The specific detection and genetic typing of trypanosomes that infect humans, mammalian reservoirs, and vectors is crucial for diagnosis and epidemiology. We utilized a PCR-RFLP assay that targeted subunit II of cytochrome oxidase and 24Sα-rDNA to simultaneously detect and discriminate six Trypanosoma cruzi discrete typing units (DTUs) and two genetic groups of Trypanosoma rangeli (KP1+/KP1-) in intestinal contents of experimentally infected Rhodnius prolixus. The PCR assays showed that in 23 of 29 (79.4%) mixed infections with the six T. cruzi DTUs and mixed infections with individual DTUs and/or groups KP1+ and KP1-, both parasites were successfully detected. In six mixed infections that involved TcIII, the TcI, TcII, TcV, and TcVI DTUs predominated to the detriment of TcIII, indicating the selection of genetic groups. Interactions between different genetic groups and vectors may lead to genetic selection over TcIII. The elimination of this DTU by the immune system of the vector appears unlikely because TcIII was present in other mixed infections (TcIII/TcIV and TcIII/KP1+). Both molecular markers used in this study were sensitive and specific, demonstrating their usefulness in a wide geographical area where distinct genotypes of these two species are sympatric. Although the cellular and molecular mechanisms that are involved in parasite-vector interactions are still poorly understood, our results indicate a dynamic selection toward specific T. cruzi DTUs in R. prolixus during mixed genotype infections.
Asunto(s)
Enfermedad de Chagas/transmisión , Insectos Vectores/parasitología , Rhodnius/parasitología , Trypanosoma cruzi/genética , Trypanosoma rangeli/genética , Animales , Brasil/epidemiología , Colombia/epidemiología , Genotipo , Humanos , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Trypanosoma cruzi/aislamiento & purificación , Trypanosoma rangeli/aislamiento & purificaciónRESUMEN
A survey of the Trypanosoma vivax genome was carried out by the genome sequence survey (GSS) approach resulting in 1,086 genomic sequences. A total of 455 high-quality GSS sequences were generated, consisting of 331 non-redundant sequences distributed in 264 singlets and 67 clusters in a total of 135.5 Kb of the T. vivax genome. The estimation of the overall G+C content, and the prediction of the presence of ORFs and putative genes were carried out using the Glimmer and Jemboss packages. Analysis of the obtained sequences was carried out by BLAST programs against 12 different databases and also using the Conserved Domain Database, InterProScan, and tRNAscan-SE. Along with the existing 23 T. vivax entries in the GenBank, the 32 putative genes predicted and the 331 non-redundant GSS sequences reported herein represent new potential markers for the development of PCRbased assays for specific diagnosis and typing of Trypanosoma vivax.
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Genoma de Protozoos , Trypanosoma vivax/genética , Trypanosoma vivax/fisiología , Algoritmos , Animales , Biología Computacional , Bases de Datos Genéticas , Bases de Datos de Ácidos Nucleicos , Etiquetas de Secuencia Expresada , Biblioteca de Genes , Genes Protozoarios/genética , Marcadores Genéticos , Sistemas de Lectura Abierta , Reacción en Cadena de la Polimerasa , Proteínas Protozoarias/fisiología , ARN de Transferencia/genética , Análisis de Secuencia de ADN , Programas InformáticosRESUMEN
The world's biodiversity, including many infectious, parasitic disease agents and their vectors whose impact on both human and animal health is significant, is largely retained in the developing countries of the tropics. Owing to the number of species involved and the relatively low-level exploration of pathogens and vectors biodiversity, several organisms are still waiting to be discovered and consequently explored in terms of genomics. Although some parasitic species of humans and animals have been studied through genomics and bioinformatics approaches, a significant number of relevant species are still to be addressed. Through the use of modern technologies, such as genomics and bioinformatics, for assessment of biodiversity and targeting tropical diseases, other relevant advantages of these initiatives for developing countries would be technology transfer and capacity building. Consequently, these initiatives could be critical to the development of the respective countries. Moreover, intra- and interhemispheric scientific collaboration should be encouraged and supported to increase the chances for success. In Brazil, the Ministry of Science and Technology has stepped forward to further such initiatives, co-supporting collaborative genomics and bioinformatics projects. The need for the establishment of working groups on genomics and bioinformatics in developing countries as well as the improvement and strengthening of collaborative research projects between developed and developing countries is discussed from our point of view. As these discussions remain open to debate, we encourage colleagues to promote further discussion on the subject.
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Biología Computacional/tendencias , Países en Desarrollo , Genómica/tendencias , Enfermedades Parasitarias/genética , Enfermedades Parasitarias/transmisión , Medicina Tropical , Organización Mundial de la Salud , Enfermedades de los Animales , Animales , Brasil , Conservación de los Recursos Naturales , Ecología , Humanos , Enfermedades Parasitarias/epidemiología , Desarrollo de Programa , Salud Pública , Medicina Tropical/estadística & datos numéricos , Medicina Tropical/tendenciasRESUMEN
The taxonomic status of Trypanosoma rangeli as well as the tools for its molecular characterization is briefly commented.
RESUMEN
BACKGROUND: The STINGRAY system has been conceived to ease the tasks of integrating, analyzing, annotating and presenting genomic and expression data from Sanger and Next Generation Sequencing (NGS) platforms. FINDINGS: STINGRAY includes: (a) a complete and integrated workflow (more than 20 bioinformatics tools) ranging from functional annotation to phylogeny; (b) a MySQL database schema, suitable for data integration and user access control; and (c) a user-friendly graphical web-based interface that makes the system intuitive, facilitating the tasks of data analysis and annotation. CONCLUSION: STINGRAY showed to be an easy to use and complete system for analyzing sequencing data. While both Sanger and NGS platforms are supported, the system could be faster using Sanger data, since the large NGS datasets could potentially slow down the MySQL database usage. STINGRAY is available at http://stingray.biowebdb.org and the open source code at http://sourceforge.net/projects/stingray-biowebdb/.
Asunto(s)
Biología Computacional/métodos , Genómica/métodos , Programas Informáticos , Flujo de Trabajo , Bases de Datos Factuales/estadística & datos numéricos , Secuenciación de Nucleótidos de Alto Rendimiento/estadística & datos numéricos , Internet , Filogenia , Reproducibilidad de los ResultadosRESUMEN
Leishmaniasis is an infectious disease caused by Leishmania species. Leishmania amazonensis is a New World Leishmania species belonging to the Mexicana complex, which is able to cause all types of leishmaniasis infections. The L. amazonensis reference strain MHOM/BR/1973/M2269 was sequenced identifying 8,802 codifying sequences (CDS), most of them of hypothetical function. Comparative analysis using six Leishmania species showed a core set of 7,016 orthologs. L. amazonensis and Leishmania mexicana share the largest number of distinct orthologs, while Leishmania braziliensis presented the largest number of inparalogs. Additionally, phylogenomic analysis confirmed the taxonomic position for L. amazonensis within the "Mexicana complex", reinforcing understanding of the split of New and Old World Leishmania. Potential non-homologous isofunctional enzymes (NISE) were identified between L. amazonensis and Homo sapiens that could provide new drug targets for development.
RESUMEN
Endosymbiont-bearing trypanosomatids have been considered excellent models for the study of cell evolution because the host protozoan co-evolves with an intracellular bacterium in a mutualistic relationship. Such protozoa inhabit a single invertebrate host during their entire life cycle and exhibit special characteristics that group them in a particular phylogenetic cluster of the Trypanosomatidae family, thus classified as monoxenics. In an effort to better understand such symbiotic association, we used DNA pyrosequencing and a reference-guided assembly to generate reads that predicted 16,960 and 12,162 open reading frames (ORFs) in two symbiont-bearing trypanosomatids, Angomonas deanei (previously named as Crithidia deanei) and Strigomonas culicis (first known as Blastocrithidia culicis), respectively. Identification of each ORF was based primarily on TriTrypDB using tblastn, and each ORF was confirmed by employing getorf from EMBOSS and Newbler 2.6 when necessary. The monoxenic organisms revealed conserved housekeeping functions when compared to other trypanosomatids, especially compared with Leishmania major. However, major differences were found in ORFs corresponding to the cytoskeleton, the kinetoplast, and the paraflagellar structure. The monoxenic organisms also contain a large number of genes for cytosolic calpain-like and surface gp63 metalloproteases and a reduced number of compartmentalized cysteine proteases in comparison to other TriTryp organisms, reflecting adaptations to the presence of the symbiont. The assembled bacterial endosymbiont sequences exhibit a high A+T content with a total of 787 and 769 ORFs for the Angomonas deanei and Strigomonas culicis endosymbionts, respectively, and indicate that these organisms hold a common ancestor related to the Alcaligenaceae family. Importantly, both symbionts contain enzymes that complement essential host cell biosynthetic pathways, such as those for amino acid, lipid and purine/pyrimidine metabolism. These findings increase our understanding of the intricate symbiotic relationship between the bacterium and the trypanosomatid host and provide clues to better understand eukaryotic cell evolution.
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Genes Protozoarios , Filogenia , Proteínas Protozoarias/genética , Simbiosis/genética , Trypanosomatina/genética , Bacterias/metabolismo , Composición de Base , Secuencia de Bases , Evolución Biológica , Leishmania major/genética , Redes y Vías Metabólicas , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Proteínas Protozoarias/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Trypanosomatina/clasificación , Trypanosomatina/metabolismo , Trypanosomatina/microbiologíaRESUMEN
BACKGROUND: The AIDS epidemic in Southern Brazil has unique features, showing co-circulation of HIV-1 subtypes C, B and recombinant forms. Florianópolis has the second highest AIDS incidence among Brazilian capitals, but limited information is available about HIV molecular epidemiology and prevalence of primary drug resistance. OBJECTIVES: To investigate the molecular epidemiology of HIV-1 in Florianópolis and to describe the prevalence of primary HIV-1 drug resistance mutations (DMRs). STUDY DESIGN: Epidemiological and clinical data from 82 untreated patients from Florianópolis (2008-2009) were analyzed. The HIV-1 subtype at envelope, protease, reverse transcriptase and integrase regions were determined by phylogenetic and bootscaning analyses and the drug resistance profile were analyzed at the Stanford HIV Drug Resistance Database. RESULTS: The most frequent HIV-1 genetic form was subtype C (65.8%) followed by mosaics BC (18.3%), subtype B (13.4%), subtype F1 (1.2%) and BCF1 recombinant (1.2%). HIV-1 subtype C and BC recombinants were much more frequent in the heterosexual exposure category, whereas subtype B was more common in the MSM exposure category. DRMs were seen in 11% of the sequences, 2.4% of them were related to PI, 5% to NRTI, 3.6% to NNRTI and 1.2% was related to INTI. CONCLUSIONS: The present study confirms the high prevalence of subtype C and BC recombinants in Santa Catarina State and revealed a significant difference in the subtype distribution among distinct virus exposure categories. This study also shows a relative high prevalence of protease/reverse transcriptase primary drug resistance mutations and corroborates the usefulness of the integrase inhibitors in southern Brazil.
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Fármacos Anti-VIH/farmacología , Farmacorresistencia Viral , Infecciones por VIH/epidemiología , Infecciones por VIH/virología , VIH-1/clasificación , VIH-1/efectos de los fármacos , Adulto , Brasil/epidemiología , Análisis por Conglomerados , Femenino , Genotipo , VIH-1/genética , VIH-1/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Filogenia , Prevalencia , Proteínas Virales/genéticaRESUMEN
Spliced leader intergenic region (SL-IR) sequences from 23 Trypanosoma rangeli strains isolated from the salivary glands of Rhodnius colombiensis, R. ecuadoriensis, R. pallescens and R. prolixus and two human strains revealed the existence of 4 genotypes with CA, GT, TA, ATT and GTAT microsatellite repeats and the presence of insertions/deletions (INDEL) and single nucleotide polymorphism (SNP) characterizing each genotype. The strains isolated from the same vector species or the same Rhodnius evolutionary line presented the same genotypes, even in cases where strains had been isolated from vectors captured in geographically distant regions. The dendrogram constructed from the SL-IR sequences separated all of them into two main groups, one with the genotypes isolated from R. prolixus and the other group containing three well defined sub-groups with the genotypes isolated from R. pallescens, R. colombiensis and R. ecuadoriensis. Random amplified polymorphic DNA (RAPD) analysis showed the same two main groups and sub-groups supporting strict T. rangeli genotypes' association with Rhodnius species. Combined with other studies, these results suggest a possible co-evolutionary association between T. rangeli genotypes and their vectors.
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Evolución Molecular , Genoma de Protozoos/genética , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Rhodnius/parasitología , Trypanosoma rangeli/genética , Animales , Evolución Biológica , ADN Intergénico/genética , ADN Protozoario/genética , Variación Genética , Genotipo , Interacciones Huésped-Parásitos , Humanos , Insectos Vectores/parasitología , Filogenia , ARN Lider Empalmado/genética , Análisis de Secuencia de ADN , Trypanosoma rangeli/clasificación , Trypanosoma rangeli/aislamiento & purificaciónRESUMEN
Diesel fuel is a potential contaminant of estuarine and mangrove areas, particularly because it is the main fuel used in small boats and larger vessels. The aim of this work was to identify genes differentially expressed in the liver of Poecilia vivipara (Guppy) exposed to 10% diesel fuel water accommodated fraction (WAF), employing the subtractive suppressive hybridization (SSH) method. The results showed 27 differentially expressed gene fragments, 12 up-regulated and 15 down-regulated. Among the up-regulated genes were CYP1A, UDPGT1a, ABCC4, Methyltransferase and Apolipoprotein A1. Down-regulated genes included Vitellogenins, C1 Inhibitor and Complement Component 3c. The identified genes are associated with different metabolic functions like biotransformation, membrane transport and immune system, indicating the susceptibility and/or molecular responses of this organism to the toxic effects elicited by diesel fuel WSF.