Characterization of new Schistosoma mansoni microsatellite loci in sequences obtained from public DNA databases and microsatellite enriched genomic libraries

In the last decade microsatellites have become one of the most useful genetic markers used in a large number of organisms due to their abundance and high level of polymorphism. Microsatellites have been used for individual identification, paternity tests, forensic studies and population genetics. Data on microsatellite abundance comes preferentially from microsatellite enriched libraries and DNA sequence databases. We have conducted a search in GenBank of more than 16,000 Schistosoma mansoni ESTs and 42,000 BAC sequences. In addition, we obtained 300 sequences from CA and AT microsatellite enriched genomic libraries. The sequences were searched for simple repeats using the RepeatMasker software. Of 16,022 ESTs, we detected 481 (3%) sequences that contained 622 microsatellites (434 perfect, 164 imperfect and 24 compounds). Of the 481 ESTs, 194 were grouped in 63 clusters containing 2 to 15 ESTs per cluster. Polymorphisms were observed in 16 clusters. The 287 remaining ESTs were orphan sequences. Of the 42,017 BAC end sequences, 1,598 (3.8%) contained microsatellites (2,335 perfect, 287 imperfect and 79 compounds). The 1,598 BAC end sequences 80 were grouped into 17 clusters containing 3 to 17 BAC end sequences per cluster. Microsatellites were present in 67 out of 300 sequences from microsatellite enriched libraries (55 perfect, 38 imperfect and 15 compounds). From all of the observed loci 55 were selected for having the longest perfect repeats and flanking regions that allowed the design of primers for PCR amplification. Additionally we describe two new polymorphic microsatellite loci.

Prevalence and genotypes of hepatitis C virus among injecting drug users from Salvador-BA, Brazil

Hepatitis C virus (HCV) is the major infectious disease agent among injecting drug users (IDUs), with seroprevalence ranging from 50-90%. In this paper, serological and virological parameters were investigated among 194 IDUs, 94 ex-IDUs and 95 non-IDUs that were sampled by the "snowball" technique in three localities renowned for both intense drug use and trafficking activities in Salvador, Brazil. The majority of the participants were male, but sex and mean age differed significantly between IDUs/ex-IDUs and non-IDUs (p < 0.05). Anti-HCV screening revealed that 35.6%, 29.8% and 5.3% of samples from IDUs, ex-IDUs and non-IDUs, respectively, were seropositive. HCV-RNA detection confirmed that the prevalence of infection was 29.4%, 21.3% and 5.3% for IDUs, ex-IDUs and non-IDUs, respectively. Genotyping analysis among IDUs/ex-IDUs determined that 76.9% were infected with genotype 1, 18.5% with genotype 3 and 4.6% with a mixed genotype; this result differed significantly from non-IDUs, where genotype 3 was the most frequent (60%), followed by genotype 1 (20%) and a mixed genotype (20%). We report a significantly higher prevalence of HCV infection in IDUs/ex-IDUs compared to the control group (p < 0.001). Although the sample size of our study was small, the differences in HCV genotype distribution reported herein for IDUs/ex-IDUs and non-IDUs warrant further investigation.

Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg

A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.

Evaluation of four molecular methods for the diagnosis of tuberculosis in pulmonary and blood samples from immunocompromised patients

The present study analysed the concordance among four different molecular diagnostic methods for tuberculosis (TB) in pulmonary and blood samples from immunocompromised patients. A total of 165 blood and 194 sputum samples were collected from 181 human immunodeficiency virus (HIV)-infected patients with upper respiratory complaints, regardless of suspicious for TB. The samples were submitted for smear microscopy, culture and molecular tests: a laboratory-developed conventional polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR) and the Gen-Probe and Detect-TB Ampligenix kits. The samples were handled blindly by all the technicians involved, from sample processing to results analysis. For sputum, the sensitivity and specificity were 100% and 96.7% for qPCR, 81.8% and 94.5% for Gen-Probe and 100% and 66.3% for Detect-TB, respectively. qPCR presented the best concordance with sputum culture [kappa (k) = 0.864)], followed by Gen-Probe (k = 0.682). For blood samples, qPCR showed 100% sensitivity and 92.3% specificity, with a substantial correlation with sputum culture (k = 0.754) and with the qPCR results obtained from sputum of the corresponding patient (k = 0.630). Conventional PCR demonstrated the worst results for sputa and blood, with a sensitivity of 100% vs. 88.9% and a specificity of 46.3% vs. 32%, respectively. Commercial or laboratory-developed molecular assays can overcome the difficulties in the diagnosis of TB in paucibacillary patients using conventional methods available in most laboratories.

Assertividade e tempo de permanência de cateteres intravenosos periféricos com inserção guiada por ultrassonografia em crianças e adolescentes

Estudo randômico e controlado que objetivou verificar se a ultrassonografia vascular (USV) aumenta a assertividade na utilização do cateter intravenoso periférico e o tempo de permanência do cateter quando comparado ao método tradicional de punção. A coleta de dados ocorreu após aprovação do mérito ético, incluindo-se no estudo crianças e adolescentes submetidos a punção intravenosa periférica guiada pela USV, constituindo o grupo USV (GUSV), ou após avaliação clínica da rede venosa, denominado grupo controle (GC). Os valores de p<0,05 foram considerados significativos. A amostra foi constituída por 382 punções, 188 (49,2%) no GUSV e 194 (50,8%) no GC, realizadas em 335 crianças. Identificou-se assertividade em 73 (71,6%) cateteres do GUSV e em 84 (71,8%) do GC (p=0,970). O tempo de permanência do cateter apresentou mediana inferior a um dia nos dois grupos (p=0,121), não havendo diferença estatisticamente significativa. Concluindo-se que a USV não influenciou os resultados das variáveis dependentes investigadas. ClinicalTrials.govNCT00930254.

Use of patients’ classification instruments: analysis of the brazilian production of knowledge

Objective To analyze the production of scientific knowledge about the use of patients’ classification instruments in care and management practice in Brazil. Method Integrative literature review with databases search in: Latin American and Caribbean Literature on Health Sciences (LILACS), Medical Literature Analysis and Retrieval System on-line (MEDLINE), Cumulative Index to Nursing and Allied Health Literature (CINAHL) and SCOPUS, between January 2002 through December 2013. Results 1,194 studies were found, 31 met the inclusion criteria. We observed a higher number of studies in the category care plans and workload (n=15), followed by the category evaluation of psychometric properties (n=14). Conclusion Brazilian knowledge production has not yet investigated some purposes of using instruments for classifying patients in professional nursing practice. The identification of unexplored areas can guide future research on the topic.


Comunidade de abelhas (Hymenoptera, Apoidea) e plantas em uma área do Agreste pernambucano, Brasil

O Agreste é uma região de transição entre floresta tropical úmida e caatinga no nordeste brasileiro. Nessa região, grande parte da vegetação nativa foi desmatada para a implantação de pastagens. Não é sabido se áreas degradadas mantém uma apifauna e flora melitófila diversificada, ou quais são associações entre abelhas e plantas que ocorrem nessas áreas. A cobertura vegetal atual é composta por pastos, vegetação ruderal e restos da vegetação nativa. Abelhas e plantas por elas visitadas foram coletadas mensalmente entre agosto de 2001 e julho de 2002, durante dois dias consecutivos entre 5h30 e 17h30. Foram coletados 1.004 indivíduos de abelhas pertencentes a 79 espécies. Apidae foi a família mais abundante e com maior riqueza de espécies (732 indivíduos e 43 espécies), seguida por Halictidae (194 indivíduos e 20 spp.), Megachilidae (47 indivíduos e 13 spp.), Colletidae (16 indivíduos e 2 spp.) e Andrenidae (15 indivíduos e 1 sp.). Foram registradas apenas três espécies de abelhas eussocais e cinco de Euglossini, dois grupos altamente diversificados nas florestas neotropicais. A ausência de abelhas sem ferrão nativas dos gêneros Plebeia, Frieseomelitta, Partamona, Scaptotrigona e Trigonisca, assim como de outras espécies de Euglossini, deve estar relacionada à falta de sítios de nidificação e à escassez de fontes de pólen e néctar nessa área degradada. Foram registradas 87 espécies de plantas melitófilas, a maioria ervas e arbustos. Árvores nativas isoladas, assim como plantas ornamentais e frutíferas cultivadas contribuem para manter parte da diversidade da comunidade de abelhas nativas.

A catalogue of types of Conopoidea, Diopsoidea, Nerioidea and Tephritoidea (Diptera, Schizophora) in the collection of the Museu de Zoologia da Universidade de São Paulo

A catalogue is provided with the type material of four superfamilies of "Acalyptrate" (Conopoidea, Diopsoidea, Nerioidea and Tephritoidea) held in the collection of the Museu de Zoologia da Universidade de São Paulo (MZUSP), São Paulo, Brazil. Concerning the taxa dealt with herein, the Diptera collection of MZUSP held 77 holotypes, 4 "allotypes" and 194 paratypes. In this paper, information about data labels, preservation and missing structures of the type specimens is given.

Efeito da formononetina (7 hidroxi, 4'metoxi isoflavona) na colonização micorrízica e crescimento do milho em solo contendo excesso de metais pesados

O excesso de metais pesados contamina o solo, exercendo impacto negativo sobre os microrganismos e ação tóxica sobre as plantas, dificultando a revegetação e reabilitação de áreas degradadas. No presente estudo, avaliou-se o efeito da aplicação do isoflavonóide formononetina (7 hidroxi, 4'metoxi isoflavona) no crescimento e absorção de metais pelo milho (Zea mayz L.) em mistura de solo com proporções crescentes de um solo contaminado que continha 16.904, 194, 219 e 836 mg dm-3 de solo de Zn, Cd, Pb e Cu, respectivamente. Esse solo foi diluído com solo não contaminado para obter misturas com 0; 2,5; 5,0; 7,5 e 10,0% p/p do solo contaminado. Nas misturas de solo, infestadas com propágulos de fungos micorrízicos, foi plantado milho, com e sem a aplicação de solução de formononetina sintética (5 mg L-1) equivalente a 400 µg kg-1 de solo. Verificou-se um efeito depressivo acentuado da elevação da contaminação do solo na colonização micorrízica e no crescimento das plantas. A formononetina estimulou a colonização nos níveis mais baixos de contaminação e exerceu efeito positivo no crescimento do milho. A absorção dos metais, em especial de Zn e Cd, aumentou com a elevação da contaminação, sendo os teores de Zn menores nas plantas com formononetina, enquanto os teores de Fe foram maiores nestas plantas até 7,5% de solo contaminado. Os resultados indicam que a formononetina reduz, de modo indireto, por meio da micorriza, os efeitos adversos do excesso de metais no solo, especialmente de Zn no milho. O favorecimento da formação de micorriza diminui a absorção de Zn e aumenta a de Fe, podendo ser este o mecanismo responsável pela ação amenizante da formononetina na toxidez de metais pesados no milho. Conclui-se que o uso da formononetina pode facilitar a revegetação de solos contaminados com metais pesados.

Influence of tree species on the herbaceous understory and soil chemical characteristics in a silvopastoral system in semi-arid northeastern Brazil

Studies from some semi-arid regions of the world have shown the beneficial effect of trees in silvopastoral systems, by promoting the formation of resource islands and increasing the sustainability of the system. No data are available in this respect for tree species of common occurrence in semi-arid Northeastern Brazil. In the present study, conducted in the summer of 1996, three tree species (Zyziphus joazeiro, Spondias tuberosa and Prosopis juliflora: ) found within Cenchrus ciliaris pastures were selected to evaluate differences on herbaceous understory and soil chemical characteristics between samples taken under the tree canopy and in open grass areas. Transects extending from the tree trunk to open grass areas were established, and soil (0-15 cm) and herbaceous understory (standing live biomass in 1 m² plots) samples were taken at 0, 25, 50, 100, 150 and 200% of the average canopy radius (average radius was 6.6 ± 0.5, 4.5 ± 0.5, and 5.3 ± 0.8 m for Z. joazeiro, P. juliflora, and S. tuberosa , respectively). Higher levels of soil C, N, P, Ca, Mg, K, and Na were found under the canopies of Z. joazeiro and P. juliflora: trees, as compared to open grass areas. Only soil Mg organic P were higher under the canopies of S. tuberosa trees, as compared to open grass areas. Herbaceous understory biomass was significantly lower under the canopy of S. tuberosa and P. juliflora trees (107 and 96 g m-2, respectively) relatively to open grass areas (145 and 194 g m-2). No herbaceous biomass differences were found between Z. joazeiro canopies and open grass areas (107 and 87 g m-2, respectively). Among the three tree species studied, Z. joazeiro was the one that presented the greatest potential for use in a silvopastoral system at the study site, since it had a larger nutrient stock in the soil without negatively affecting herbaceous understory biomass, relatively to open grass areas.

Manganese accumulation and its relation to "eucalyptus shoot blight in the Vale do Rio Doce"

Eucalyptus Shoot Blight in the Vale do Rio Doce (ESBVRD) is an anomaly that leads to reduced growth and, in more extreme cases, to death of eucalyptus plants. Initially diagnosed in plantations in the region of the Vale do Rio Doce, in the State of Minas Gerais, Brazil, this problem has also been found in plantations in other regions of the country and even in other countries. Although the symptoms of this anomaly are well-known, its causes are not yet understood. The aim of this study was to evaluate the cause-effect relationship between accumulation of manganese (Mn) in eucalyptus clones and ESBVRD. Characterization of the environment in areas of greater occurrence of this problem in regard to soil, climate and fluctuation of the water table was undertaken in eucalyptus plantations of the Celulose Nipo-brasileira S.A. (Cenibra) company in the region of the Vale do Rio Doce. Plant tissues were sampled in two situations. In the first situation, diagnosis occurred in the initial phase of the anomaly in clones with differentiated tolerance to the problem; in the second situation, diagnosis was made in a single clone, considered to be sensitive, in two time periods - in the phase with the strong presence of symptoms and in the recovery phase, in areas of occurrence and in areas of escape from the problem. The most ESBVRD-sensitive clone showed much higher (4.8 times higher) leaf Mn contents than more tolerant clones. In plants with the anomaly, Mn leaf contents were greater than 3,070 mg kg-1, much greater than the quantity found in those without the anomaly (734 mg kg-1). In the period in which the symptoms began to wane, there was a sharp decline in leaf Mn contents, from 2,194 to 847 mg kg-1. Manganese content in the above ground part and plant litter (44.4 g ha-1) in the area of occurrence of the anomaly was three times greater than that found in these same components (14.1 g ha-1) in the area of absence of the symptom. Based on the evidence found, such as the existence of environmental conditions favorable to high Mn availability to the plants in the areas of greatest incidence of ESBVRD, greater uptake of Mn in sensitive clones and in plants with symptoms, and a synchronism between the intensity of symptoms of ESBVRD and leaf Mn contents, it may be inferred that temporary excess of Mn in eucalyptus plants is closely related to ESBVRD.

Consumo e custo da alimentação para recuperação da atividade ovariana luteal cíclica de vacas mestiças leiteiras com anestro

O objetivo do trabalho foi verificar o consumo e o custo de alimentos para recuperação da atividade ovariana luteal cíclica (AOLC) em vacas mestiças Holandês x Zebu com anestro. Foram usadas 18 vacas, não-gestantes, não-lactantes, magras, apresentando ovários sem função luteal, de tamanho normal e sem folículos palpáveis na superfície. Doze animais permaneceram em confinamento e receberam alimentação para ganho de peso até o reinício da AOLC. Os seis animais restantes, constituindo o grupo controle, receberam alimentação de mantença para o baixo peso apresentado e permaneceram em anestro durante o período experimental. A AOLC foi avaliada pela concentração de progesterona no soro sangüíneo (coleta de sangue a cada sete dias), pelo exame dos ovários por palpação retal a cada 12 dias, e observação visual dos sinais do estro três vezes ao dia. A recuperação da AOLC nas vacas com anestro exigiu um consumo médio de 722,4 kg de matéria seca, 50,4 kg de proteína bruta e 402,4 kg de NDT, relativos à ingestão média de 2.374,2 kg de volumoso e 194,1 kg de concentrado. O custo desses alimentos somado à perda estimada da produção de leite numa vaca de 3.000 litros de leite/lactação, provocada pelo prolongamento do intervalo de partos, foi equivalente a 1.364,2 litros de leite (preço recebido pelo produtor = R$ 0,20/litro). Esse custo elevado da recuperação do anestro onera o custo final da produção de leite, tendo em vista a alta incidência de anestro nos rebanhos brasileiros.

Tomato yield and potassium concentrations in soil and in plant petioles as affected by potassium fertirrigation

Tomato (Lycopersicon esculentum Mill.) cv. Santa Clara was grown on a silt clay soil with 46 mg dm-3 Mehlich 1 extractable K, to evaluate the effects of trickle-applied K rates on fruit yield and to establish K critical concentrations in soil and in plant petioles. Six potassium rates (0, 48, 119, 189, 259 and 400 kg ha-1 K) were applied in a randomized complete block design with four replications. Soil and plant K critical levels were determined at two plant growth stages (at the beginning of the second and fourth cluster flowering). Total, marketable and weighted yields increased with K rates, reaching their maximum of 86.4, 73.4, and 54.9 ton ha-1 at 198, 194, and 125 kg ha-1 K , respectively. At the first soil sampling date K critical concentrations in the soil associated with K rates for maximum marketable and weighted yields were 92 and 68 mg dm-3, respectively. Potassium critical concentrations in the dry matter of the petioles sampled by the beginning of the second and fourth cluster flowering time, associated with maximum weighted yield, were 10.30 and 7.30 dag kg-1, respectively.

Desenvolvimento ponderal de bezerras desmamadas em pastejo de Brachiaria decumbens com suplementação protéica e energética

Foi avaliado o efeito da suplementação a pasto, com concentrados fornecidos durante o ano (períodos de seca e águas), sobre o desenvolvimento ponderal de bezerras desmamadas. Foram utilizadas 36 bezerras da raça Santa Gertrudis, com média inicial de idade de 290 dias, média de peso vivo de 194 kg, mantidas em pasto de Brachiaria decumbens, distribuídas em um delineamento inteiramente casualizado, com dois tratamentos e 18 repetições. Foi oferecida mistura mineral o ano todo, ou suplemento com maior nível de proteína degradável na seca, e não-degradável nas águas. Os resultados obtidos para a média diária de ganho de peso vivo indicaram uma diferença significativa entre os tratamentos. O ganho de peso vivo, durante o ano, para o lote testemunha, foi de 0,504 kg animal-1 dia-1 e, para o lote suplementado foi de 0,561 kg. No período de seca, o ganho de peso vivo foi significativamente maior para os animais suplementados e, no período das águas não ocorreu diferença. A suplementação a pasto não apresentou vantagens bioeconômicas para se recomendar o seu uso. Nos dois tratamentos as bezerras alcançaram média de peso limite, para primeira cobertura, com idade próxima a 14 meses.

Atividade isoenzimática em plantas de trigo infectadas com o vírus SBWMV

O objetivo deste trabalho foi elucidar a atividade e a expressão isoenzimática das esterases, das peroxidases e das aspartato aminotransferases em função da infecção de plantas de trigo pelo Soil-borne wheat mosaic virus (SBWMV). Foram analisadas, aos 45 dias após a emergência, quatro cultivares e uma linhagem de trigo, com diferentes níveis de resistência ao SBWMV: BRS Guabiju, BRS 194, BRS 179, BR 23 e PF 980524. De modo geral, ocorreram diferenças qualitativas e quantitativas intra e interpopulacional, quando comparadas plantas assintomáticas e sintomáticas ao SBWMV. Para o sistema esterase, nove padrões de bandas foram determinados e para peroxidase e aspartato aminotransferase foram detectados três padrões de bandas, para ambas as condições. Padrões eletroforéticos foram observados para plantas infectadas, quando comparadas com as não infectadas, destacando-se a atividade da esterase, o que permitiu identificar com maior precisão o estado metabólico e diferenciado das células.