A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni

Schistosomiasis is a major neglected tropical disease caused by trematodes from the genus Schistosoma. Because schistosomes exhibit a complex life cycle and numerous mechanisms for regulating gene expression, it is believed that spliced leader (SL) trans-splicing could play an important role in the biology of these parasites. The purpose of this study was to investigate the function of trans-splicing in Schistosoma mansoni through analysis of genes that may be regulated by this mechanism and via silencing SL-containing transcripts through RNA interference. Here, we report our analysis of SL transcript-enriched cDNA libraries from different S. mansoni life stages. Our results show that the trans-splicing mechanism is apparently not associated with specific genes, subcellular localisations or life stages. In cross-species comparisons, even though the sets of genes that are subject to SL trans-splicing regulation appear to differ between organisms, several commonly shared orthologues were observed. Knockdown of trans-spliced transcripts in sporocysts resulted in a systemic reduction of the expression levels of all tested trans-spliced transcripts; however, the only phenotypic effect observed was diminished larval size. Further studies involving the findings from this work will provide new insights into the role of trans-splicing in the biology of S. mansoni and other organisms. All Expressed Sequence Tags generated in this study were submitted to dbEST as five different libraries. The accessions for each library and for the individual sequences are as follows: (i) adult worms of mixed sexes (LIBEST_027999: JZ139310 - JZ139779), (ii) female adult worms (LIBEST_028000: JZ139780 - JZ140379), (iii) male adult worms (LIBEST_028001: JZ140380 - JZ141002), (iv) eggs (LIBEST_028002: JZ141003 - JZ141497) and (v) schistosomula (LIBEST_028003: JZ141498 - JZ141974).

A serological, parasitological and clinical evaluation of untreated Chagas disease patients and those treated with benznidazole before and thirteen years after intervention

The etiological treatment of Chagas disease is recommended for all patients with acute or recent chronic infection, but controversies remain regarding the benefit of chemotherapy and interpretations of the parasitological cure after etiological treatment. This study compares the laboratory and clinical evaluations of Chagas disease patients who were diagnosed 13 years earlier. Fifty-eight Chagas disease patients (29 treated with benznidazole and 29 untreated) were matched at the time of treatment based on several variables. Conventional serology revealed the absence of seroconversion in all patients. However, lower serological titres were verified in the treated group, primarily among patients who had the indeterminate form of the disease. Haemoculture performed 13 years after the intervention was positive for 6.9% and 27.6% of the treated and untreated patients, respectively. Polymerase chain reaction tests were positive for 44.8% and 13.8% of the treated and untreated patients, respectively. Patients who presented with the indeterminate form of the disease at the beginning of the study exhibited less clinical progression (17.4%) compared with the untreated group (56.5%). Therefore, this global analysis revealed that etiological treatment with benznidazole may benefit patients with respect to the clinical progression of Chagas disease and the prognosis, particularly when administered to patients with the indeterminate form of the disease.

Species composition and seasonal abundance of sandflies (Diptera: Psychodidae: Phlebotominae) in coffee agroecosystems

The composition and seasonal occurrence of sandflies were investigated in coffee agroecosystems in the Soconusco region of Chiapas, Mexico. Insect sampling was performed on three plantations located at different altitudes: Finca Guadalupe Zajú [1,000 m above sea level (a.s.l.)], Finca Argovia (613 m a.s.l.) and Teotihuacán del Valle (429 m a.s.l.). Sandflies were sampled monthly from August 2007-July 2008 using three sampling methods: Shannon traps, CDC miniature light traps and Disney traps. Sampling was conducted for 3 h during three consecutive nights, beginning at sunset. A total of 4,387 sandflies were collected during the course of the study: 2,718 individuals in Finca Guadalupe Zajú, 605 in Finca Argovia and 1,064 in Teotihuacán del Valle. The Shannon traps captured 94.3% of the total sandflies, while the CDC light traps and Disney traps captured 4.9% and 0.8%, respectively. More females than males were collected at all sites. While the number of sandflies captured was positively correlated with temperature and relative humidity, a negative correlation was observed between sandfly numbers and rainfall. Five species of sandflies were captured: Lutzomyia cruciata , Lutzomyia texana , Lutzomyia ovallesi , Lutzomyia cratifer / undulata and Brumptomyia sp. Lu. cruciata , constituting 98.8% of the total, was the most abundant species. None of the captured sandflies was infected with Leishmaniaspp.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

Studies of genotoxicity and mutagenicity of nitroimidazoles: demystifying this critical relationship with the nitro group

Nitroimidazoles exhibit high microbicidal activity, but mutagenic, genotoxic and cytotoxic properties have been attributed to the presence of the nitro group. However, we synthesised nitroimidazoles with activity against the trypomastigotes of Trypanosoma cruzi, but that were not genotoxic. Herein, nitroimidazoles (11-19) bearing different substituent groups were investigated for their potential induction of genotoxicity (comet assay) and mutagenicity (Salmonella/Microsome assay) and the correlations of these effects with their trypanocidal effect and with megazol were investigated. The compounds were designed to analyse the role played by the position of the nitro group in the imidazole nucleus (C-4 or C-5) and the presence of oxidisable groups at N-1 as an anion receptor group and the role of a methyl group at C-2. Nitroimidazoles bearing NO2 at C-4 and CH3 at C-2 were not genotoxic compared to those bearing NO2 at C-5. However, when there was a CH3 at C-2, the position of the NO2 group had no influence on the genotoxic activity. Fluorinated compounds exhibited higher genotoxicity regardless of the presence of CH3 at C-2 or NO2 at C-4 or C-5. However, in compounds 11 (2-CH3; 4-NO2; N-CH2OHCH2Cl) and 12 (2-CH3; 4-NO2; N-CH2OHCH2F), the fluorine atom had no influence on genotoxicity. This study contributes to the future search for new and safer prototypes and provide.

Clostridium difficile ribotypes in humans and animals in Brazil

Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR) ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileribotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR ribotypes. Among toxigenic strains, PCR ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.

Evolução da demanda ao Curso de Graduação em Enfermagem da Universidade Federal de São Carlos

O estudo analisa a evolução da procura pelo Curso de Graduação em Enfermagem da Universidade Federal de São Carlos (UFSCar), desde o 1º vestibular em 1977 até 1996. Nessa análise serão utilizados comparativamente os dados dos Cursos da Universidade de São Paulo (USP) e da Escola Paulista de Medicina, atualmente denominada Uiversidade Federal de São Paulo (UNIFESP). A crise econômica e do setor da saúde influíram significativamente na queda súbita da procura pelos Cursos na década de 80. Em 1990 o vestibular da UFSCar passou a ser realizado pela Fundação Universitária para o Vestibular (FUVEST). Nesse ano a relação candidato/ vaga atingiu seu nível mais baixo, desde 1977. A partir de 1991, a tendência de retomada do crescimento econômico, e a implantação do Sistema Único de Saúde (SUS) determinaram o crescimento da demanda. Dos três cursos da carreira da Enfermagem na FUVEST, o da UFSCar que é o único do interior do Estado, ainda é o mais procurado em 3ª opção, apesar da procura em 1ª opção ter passado de 2,0 em 1990 para 6,0 em 1995, caindo para 4,6 em 1996. A pouca procura em 1990; os critérios de corte e o procedimento de chamada dos candidatos classificados concorreram para que das 30 vagas oferecidas fossem repreenchidas apenas 15, mesmo com dois vestibulares.

Avaliação do estado nutricional de crianças menores de 10 anos no município de Ferros, Minas Gerais

A sociedade brasileira vivencia, além da desnutrição e fome, problemas relacionados à obesidade - a transição nutricional. Este trabalho é um estudo transversal com amostra de 1322 crianças menores de 10 anos, residentes no município de Ferros, Minas Gerais, e cadastradas no SISVAN. Observou-se que 20,7% das crianças apresentaram alguma alteração nutricional (10,1% risco nutricional, 3,8% desnutrição e 6,7% sobrepeso). Fatores de risco para o desenvolvimento de desnutrição foram pesquisados: baixo peso ao nascer, RP=3,57, IC 95% (1,96-6,52); baixa estatura RP=19,36, IC 95% (11,53-32,52); aleitamento materno ausente RP=2,23, IC 95% (1,19-4,18); renda familiar de até R$ 95, RP = 2,39, IC 95% (1,10-5,16). Destaca-se maior prevalência de sobrepeso em relação à desnutrição. Enquanto a população rural apresentou maior prevalência de desnutrição e risco nutricional, a urbana destacou-se pelo sobrepeso. Esses resultados são importantes para mostrar que intervenções nutricionais são necessárias e devem considerar os problemas específicos apontados.

Representações sociais de enfermeiros acerca do estresse laboral em um serviço de urgência

Neste estudo objetivou-se apreender as representações de enfermeiros sobre o seu trabalho em serviço de urgência e sua relação com o estresse. Trata-se de um estudo exploratório descritivo e de abordagem qualitativa. Utilizou-se a Teoria das Representações Sociais. O cenário da pesquisa foi um hospital público de referência em urgência do município de Natal/RN. A amostra foi composta por dez enfermeiros. Para a análise dos dados, utilizou-se a técnica de análise do conteúdo. Os resultados apontam a relação das representações sociais do trabalho com o estresse dos enfermeiros, como um fenômeno complexo e multifacetado, contributivo de doenças e desgaste físico, emocional e mental.

Evaluation of the Primary Care in leprosy control: proposal of an instrument for users

Objective Developing an instrument to evaluate the performance of primary health care in the leprosy control actions, from the perspective of users and do the face and content validation. Method This is a methodological study carried out in four stages: development of instrument, face and content validation, pre-test, and analysis of test-retest reliability. Results The initial instrument submitted to the judgment of 15 experts was composed of 157 items. The face and content validation and pre-test of instrument were essential for the exclusion of items and adjustment of instrument to evaluate the object under study. In the analysis of test-retest reliability, the instrument proved to be reliable. Conclusion The instrument is considered adequate, but further studies are needed to test the psychometric properties.


As abelhas eussociais (Hymenoptera, Apidae) visitantes florais em um ecossistema de dunas continentais no médio Rio São Francisco, Bahia, Brasil

Highly eusocial bees (Hymenoptera, Apidae) flower visitors in a continental sand dune ecosystem from the medium São Francisco River, Bahia, Brazil. A community of highly eusocial bees in sand dunes, covered with caatinga vegetation, in the medium São Francisco River, Bahia (10º47' 37"S and 42º49' 25"W) was studied. The local climate is semi arid and hot, with mean temperature of 25.7 ºC and annual precipitation of 653.8 mm. Censuses took place every two months, from February to December of 2000. The bees were sampled on flowers with entomological nets, from 6:00 a.m. to 5:00 p.m. A total of 2,147 individuals of eight species of Apinae were found, of which Apis mellifera Linnaeus (40.2%), Trigona spinipes (Fabricius) (28.7%) and Frieseomelitta silvestri languida Moure (14.7%) were the predominant species. The diversity was H' = 1.53 and the evenness E' = 0.73. The bees were active during the whole year, but there was a significant variation in the monthly abundance of individuals (c2= 799.55; df= 35; p<0.0001). The daily activity was greater between 6:00 a.m. and 8:00 a.m. The low bee diversity observed is a consequence of the low richness of botanical species and of the small amount of sites for the bees' nests. The community of highly eusocial bees from the dunes presents organization patterns similar to those observed in other caatinga areas, albeit with some particularities.

Análise de fauna e flutuação populacional de Carabidae e Staphylinidae (Coleoptera) em sistemas de plantio direto e convencional

O objetivo deste estudo foi analisar a fauna de carabídeos e estafilinídeos por meio de vários índices e obter a flutuação populacional das espécies dominantes em fragmento florestal e cultura de soja/milho sob sistemas de plantio direto e convencional. As amostragens dos coleópteros foram realizadas no período de novembro/2004 a abril/2007 em Guaíra, São Paulo, sendo quinzenal durante o período de safra e mensal nas entressafras. Para a obtenção das amostras utilizou-se armadilhas de solo distribuídas em dois transectos de 200 m de comprimento, sendo 100 m na cultura e 100 m no fragmento. A fauna foi caracterizada pelos índices de diversidade e equitabilidade e pela abundância. No sistema de plantio direto capturou-se maior número de espécies de carabídeos e estafilinídeos e os índices de equitabilidade e diversidade indicaram que a comunidade desses besouros mostrou-se melhor estruturada quando comparado com a observada no sistema de plantio convencional. Entre as espécies de carabídeos destacou-se Abaris basistriatus por ter se caracterizado como dominante na cultura e fragmento florestal das duas áreas experimentais. As espécies Scarites sp. 4 e A. basistriatus geralmente apresentaram picos populacionais quando a cultura de soja contava menos de 30 dias da implantação, as demais espécies apresentaram picos populacionais que ocorreram em períodos variados das safras. A precipitação pluvial foi o fator meteorológico que obteve a maioria dos coeficiente positivos em seguida o fator temperatura mínima.

Survey of potential sharpshooter and spittlebug vectors of Xylella fastidiosa to grapevines at the São Francisco River Valley, Brazil

Survey of potential sharpshooter and spittlebug vectors of Xylella fastidiosa to grapevines at the São Francisco River Valley, Brazil. Pierce's disease of grapevines, caused by Xylella fastidiosa, is a serious problem in some regions of North America, not yet reported in Brazil. In this study, a survey of potential sharpshooter (Hemiptera, Cicadellidae, Cicadellinae) and spittlebug (Hemiptera, Cercopidae) vectors of X. fastidiosa was conducted in vineyards at the São Francisco River Valley, a major grape growing region in Brazil. Four vineyards of Vitis vinifera L. were sampled fortnightly from June/2005 to June/2007, using yellow sticky cards, each placed at two different heights (45 cm aboveground and 45 cm above the crop canopy) in 10 sampling localities. A total of 4,095 specimens of sharpshooters were collected, nearly all from 3 Proconiini species, Homalodisca spottii Takiya, Cavichioli & McKamey, 2006 (96.8% of the specimens), Tapajosa fulvopunctata (Signoret, 1854) (3.1%), and Tretogonia cribrata Melichar, 1926 (1 specimen). Hortensia similis (Walker, 1851) (2 specimens) was the only Cicadellini species. Only 1 cercopid specimen, belonging to Aeneolamia colon (Germar, 1821), was trapped. Even though they are not considered potential Xylella vectors, 2 Gyponini leafhoppers were collected: Curtara samera DeLong & Freytag, 1972 (11 specimens) and Curtara inflata DeLong & Freytag, 1976 (1 specimen). Homalodisca spottii was observed feeding and mating on green branches of grapevines, in addition to egg masses. Because of its prevalence on the crop canopy, occurrence throughout the year (with peaks from February to August), and ability to colonize grapevines, H. spottii could be an important vector if a X. fastidiosa strain pathogenic to grapevines becomes introduced at the São Francisco River Valley.

Viabilidade do uso da adubação verde nos agroecossistemas da região de São Carlos, SP

De modo geral, os sistemas de produção agrícola têm levado os agroecossistemas a uma grande instabilidade, acarretando forte degradação ambiental. Este trabalho visa estudar o potencial da utilização de adubação verde na região de São Carlos (SP) como uma técnica complementar aos sistemas de produção ora existentes, buscando alcançar um incremento qualitativo de seus agroecossistemas. A delimitação da região estudada baseou-se em características climáticas, geomorfológicas e administrativas, a partir de informações oriundas de mapas temáticos, manipulados por meio de um sistema de informações geográficas (SIG). A unidade de planejamento agroecológico para a região de São Carlos é constituída pelos municípios de Analândia, Itirapina, Santa Maria da Serra, São Carlos, São Pedro e Torrinha. Na região, descreveram-se o clima e os tipos de solo, tendo sido estes últimos agrupados de acordo com suas respectivas texturas e fertilidades. A partir disto, gerou-se um mapa com as zonas edáficas, predominantemente álicas e de textura arenosa. Paralelamente, foram listadas espécies de adubo verde com ecofisiologia compatível com as características edafoclimáticas encontradas, agrupando tais espécies em três classes: exigentes, rústicas e muito rústicas. Finalmente, elaborou-se um mapa com as zonas de cultivo de cada grupo de espécies. Concluiu-se que a adubação verde é uma técnica com potencial para melhorar a qualidade dos agroecossistemas da região de São Carlos.

Caracterização de podzólicos amarelos irrigados e não irrigados do submédio São Francisco

Investigações micromacromorfológicas, físicas, químicas e mineralógicas foram realizadas em solos Podzólicos Amarelos na região do submédio São Francisco, município de Petrolina, estado de Pernambuco. O objetivo do estudo foi caracterizar e identificar possíveis diferenciações nos solos, em função de seus posicionamentos no terreno, pela ação conjugada dos relevos de superfície e subsuperfície, subordinadas ou não a cinco anos de manejo com irrigação em cultura de manga. Foram utilizados dados de quatro perfis de solos, sendo dois em área irrigada e dois em outra contígua que apenas tinha sido recentemente desmatada. Observou-se que os solos não foram significantemente alterados em suas características morfológicas e mineralógicas pelo manejo com irrigação. Algumas características morfológicas, como presença de fragipãs e mosqueamentos, parecem estar relacionadas com a movimentação e acumulação de água subordinadas à conjugação dos relevos de superfície e subsuperfície. Modificações nas características químicas foram observadas, principalmente relativas à alta salinidade e sodicidade, especialmente nos horizontes mais profundos dos perfis da subárea não irrigada. Tais modificações também parecem relacionadas com a dinâmica da água subordinada ao relevo. Uma vez que alguns efeitos nocivos da irrigação foram observados na área contígua, que não estava sendo utilizada, ressalta-se a necessidade de sempre extrapolar a avaliação do impacto ambiental da irrigação além dos limites das áreas dos projetos. Também foi observado que a cimentação e a impermeabilidade nos horizontes mais profundos são, provavelmente, devidas à presença de micropãs formados por argilas iluviais.