RESUMEN
This work presents the first identification of putative odorant-binding proteins (OBPs) from a member of the Pentatomidae, i.e. the brown stink bug Euschistus heros (Fabricius), an important pest of soybean in Brazil. Antennae from both sexes of E. heros adults (12 days old and unmated) were used to construct a cDNA library, from which two transcripts encoding putative E. heros OBPs (EherOBPs) were identified. The expression levels of EherOBP1 and EherOBP2 were found to be higher in male antennae than in female and there was difference in expression in legs, wings, and abdomens of the two sexes. The histolocalization of EherOBP1 and EherOBP2 transcripts in antennae also showed a sexual dimorphism in the chemoreception system, with different expression sites in the antennal segments between males and females, occurring predominantly at the base of the sensillum. The implications of these findings for stink bug chemoreception are discussed.
Asunto(s)
Heterópteros/química , Receptores Odorantes/análisis , Animales , Brasil , Femenino , Masculino , Receptores Odorantes/metabolismo , Glycine maxRESUMEN
Approximately 50% of the predicted protein-coding genes of the Trypanosoma cruzi CL Brener strain are annotated as hypothetical or conserved hypothetical proteins. To further characterize these genes, we generated 1161 open-reading frame expressed sequence tags (ORESTES) from the mammalian stages of the VL10 human strain. Sequence clustering resulted in 435 clusters, consisting of 339 singletons and 96 contigs. Significant matches to the T. cruzi predicted gene database were found for ~94% contigs and ~69% singletons. These included genes encoding surface proteins, known to be intensely expressed in the parasite mammalian stages and implicated in host cell invasion and/or immune evasion mechanisms. Among 151 contigs and singletons with similarity to predicted hypothetical protein-coding genes and conserved hypothetical protein-coding genes, 83% showed no match with T. cruzi EST and/or proteome databases. These ORESTES are the first experimental evidence that the corresponding genes are in fact transcribed. Sequences with no significant match were searched against several T. cruzi and National Center for Biotechnology Information non-redundant sequence databases. The ORESTES analysis indicated that 124 predicted conserved hypothetical protein-coding genes and 27 predicted hypothetical protein-coding genes annotated in the CL Brener genome are transcribed in the VL10 mammalian stages. Six ORESTES annotated as hypothetical protein-coding genes showing no match to EST and/or proteome databases were confirmed by Northern blot in VL10. The generation of this set of ORESTES complements the T. cruzi genome annotation and suggests new stage-regulated genes encoding hypothetical proteins.
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Etiquetas de Secuencia Expresada , Sistemas de Lectura Abierta , Proteínas Protozoarias/genética , Trypanosoma cruzi/genética , Animales , Genoma de Protozoos , HumanosRESUMEN
AIMS: We performed the first characterization of the microbiota associated with the reef coral Mussismilia braziliensis by means of a culture-independent approach. METHODS AND RESULTS: The main groups were Proteobacteria, Cyanobacteria and unclassified bacteria according to the 16S rDNA libraries. Most of the sequences of the mucus of healthy and diseased M. braziliensis did not find close matches in GenBank (i.e. >97% 16S rDNA similarity). Most of the sequences of seawater and mucus of healthy coral fell into tight clusters (17 and 15 clusters respectively). In contrast, most of the sequences of mucus of diseased coral did not form clusters. The rarefaction curves indicate saturation in the recovery of higher taxa (approximately 40 phyla). However, the number of species in the coral mucus (n = 130-170) and seawater (n = 170) did not reach a plateau. CONCLUSIONS: The coral microbiota encompasses several potentially novel species and higher taxa. The microbiota of M. braziliensis appears to be species-specific. Diseased coral may have provided a suitable place for colonization by opportunistic bacteria, resulting in a greater bacterial diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: The first study on the diversity of the microbiota of the endemic and endangered of extinction coral M. braziliensis.
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Antozoos/microbiología , Bacterias/clasificación , Ecosistema , Agua de Mar/microbiología , Animales , Océano Atlántico , Bacterias/genética , Biodiversidad , Brasil , ADN Bacteriano/genética , Biblioteca de Genes , Datos de Secuencia Molecular , Proteobacteria/clasificación , Proteobacteria/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
AIMS: The aim of this study was to determine the genetic variability in Aspergillus flavus populations from Brazil nut and cashew and develop a polymerase chain reaction (PCR) detection method. METHODS AND RESULTS: Chomatography analysis of 48 isolates identified 36 as aflatoxigenic (75%). One hundred and forty-one DNA bands were generated with 11 random amplified polymorphic DNA (RAPD) primers and analysed via unweighted pair group analysis, using arithmetic means (UPGMA). Isolates grouped according to host, with differentiation of those from A. occidentale also according to geographical origin. Aspergillus flavus-specific PCR primers ASPITSF2 and ASPITSR3 were designed from ribosomal DNA internal transcribed spacers (ITS 1 and 2), and an internal amplification control was developed, to prevent false negative results. Specificity to only A. flavus was confirmed against DNA from additional aspergilli and other fungi. CONCLUSIONS: RAPD-based characterization differentiated isolates according to plant host. The PCR primer pair developed showed specificity to A. flavus, with a detection limit of 10 fg. SIGNIFICANCE AND IMPACT OF THE STUDY: Genetic variability observed in A. flavus isolates from two Brazilian agroecosystems suggested reproductive isolation. The PCR detection method developed for A. flavus represents progress towards multiplex PCR detection of aflatoxigenic and nonaflatoxigenic strains in Hazard Analysis Critical Control Point systems.
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Aspergillus flavus/aislamiento & purificación , ADN Ribosómico/análisis , Técnicas de Tipificación Micológica , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Aflatoxinas/análisis , Aflatoxinas/biosíntesis , Aflatoxinas/genética , Anacardium/microbiología , Aspergillus flavus/genética , Bertholletia/microbiología , Brasil , ADN de Hongos/análisis , ADN Ribosómico/genéticaRESUMEN
SUMMARY: A web-based application to analyze protein amino acids conservation-Consensus Sequence (ConSSeq) is presented. ConSSeq graphically represents information about amino acid conservation based on sequence alignments reported in homology-derived structures of proteins. Beyond the relative entropy for each position in the alignment, ConSSeq also presents the consensus sequence and information about the amino acids, which are predominant at each position of the alignment. ConSSeq is part of the STING Millennium Suite and is implemented as a Java Applet. AVAILABILITY: http://sms.cbi.cnptia.embrapa.br/SMS/STINGm/consseq/, http://trantor.bioc.columbia.edu/SMS/STINGm/consseq/, http://mirrors.rcsb.org//SMS/STINGm/consseq/, http://www.es.embnet.org/SMS/STINGm/consseq/ and http://www.ar.embnet.org/SMS/STINGm/consseq/
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Bases de Datos de Proteínas , Internet , Proteínas/química , Análisis de Secuencia de Proteína/métodos , Homología de Secuencia de Aminoácido , Programas Informáticos , Interfaz Usuario-Computador , Aminoácidos/química , Secuencia Conservada , Conformación Proteica , Proteínas/análisis , Proteínas/clasificación , Alineación de Secuencia/métodos , Relación Estructura-ActividadRESUMEN
Arachis hypogaea L., commonly known as the peanut or groundnut, is an important and widespread food legume. Because the crop has a narrow genetic base, genetic diversity in A. hypogaea is low and it lacks sources of resistance to many pests and diseases. In contrast, wild diploid Arachis species are genetically diverse and are rich sources of disease resistance genes. The majority of known plant disease resistance genes encode proteins with a nucleotide binding site domain (NBS). In this study, degenerate PCR primers designed to bind to DNA regions encoding conserved motifs within this domain were used to amplify NBS-encoding regions from Arachis spp. The Arachis spp. used were A. hypogaea var. Tatu and wild species that are known to be sources of disease resistance: A. cardenasii, A. duranensis, A. stenosperma and A. simpsonii. A total of 78 complete NBS-encoding regions were isolated, of which 63 had uninterrupted ORFs. Phylogenetic analysis of the Arachis NBS sequences derived in this study and other NBS sequences from Arabidopsis thaliana, Medicago trunculata, Glycine max, Lotus japonicus and Phaseolus vulgaris that are available in public databases This analysis indicates that most Arachis NBS sequences fall within legume-specific clades, some of which appear to have undergone extensive copy number expansions in the legumes. In addition, NBS motifs from A. thaliana and legumes were characterized. Differences in the TIR and non-TIR motifs were identified. The likely effect of these differences on the amplification of NBS-encoding sequences by PCR is discussed.
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Arachis/clasificación , Arachis/genética , Genes de Plantas , Filogenia , Arabidopsis/clasificación , Arabidopsis/genética , Secuencia de Bases , Cartilla de ADN , Inmunidad Innata , Datos de Secuencia Molecular , Enfermedades de las Plantas/genética , Reacción en Cadena de la PolimerasaRESUMEN
The prevalence rates of hypertension among adult (ages 18-74) Mexican Americans, Cuban Americans, and Puerto Ricans were estimated using data from the 1982-84 Hispanic Health and Nutrition Examination Survey (HHANES). Hypertension is defined as diastolic greater than or equal to 90 mm Hg, or systolic greater than or equal to 140 mm Hg, or currently taking antihypertensive medication. Among Mexican Americans in the Southwestern United States, 16.8 percent of the males and 14.1 percent of the females were found to be hypertensive. Among Cuban Americans in Dade County, Florida 22.8 percent of the males and 15.5 percent of the females were hypertensive. Among Puerto Ricans in the New York City area 15.6 percent of the males and 11.5 percent of the females were hypertensive. The age-adjusted rates are significantly lower than comparable rates for Whites and Blacks as measured in the second National Health and Nutrition Examination Survey (NHANES II), 1976-80. Control of hypertension in the HHANES populations fall short of the 1990 Objectives for the Nation established by the US Public Health Service 60 percent (34 percent controlled Mexican American hypertensives, 27.8 percent controlled Cuban American hypertensives, and 29 percent controlled Puerto Rican hypertensives.
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Hispánicos o Latinos , Hipertensión/epidemiología , Adulto , Anciano , Presión Sanguínea , Cuba/etnología , Femenino , Encuestas Epidemiológicas , Humanos , Hipertensión/etnología , Masculino , México/etnología , Persona de Mediana Edad , Puerto Rico/etnología , Estados UnidosRESUMEN
The risk of fatal hemorrhage may limit the completeness of resection in hepatic malignancies and in vascular extensions of Wilms' tumors. We have used Ein's technique of deep hypothermia (average 17 degrees C) with cardiac arrest (average 39 minutes) and exsanguination in performing five hepatic and two intravenous Wilms' tumor resections. The initial hepatic resection takes less than 15 minutes to perform in a bloodless field and the specimen is immediately examined by frozen section for determination of adequacy of margin. Additional resection is easily performed. Of four trisegmentectomies and one left lobectomy, two required additional resections. Mattress sutures were used to control hemorrhage during recirculation. One patient died from bleeding and cardiac decompensation and another from recurrence of tumor. The Wilms' tumors extended from the iliac vein into the right atrium in one child and from the right renal vein to the right atrium with extensions into the hepatic and lumbar veins in another. After nephrectomy, the atria and inferior vena cava were opened and the tumor extracted under direct vision. Both patients are well.