PCR with Paracoccidioides brasiliensis specific primers: potential use in ecological studies

The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis genes. This fungus as well as several other fungi, were grown and their DNA obtained by mechanical disruption and a phenol chloroform isoamylalcohol-based purification method. The DNA served for a PCR reaction that employed specific primers from two P. brasiliensis genes that codify for antigenic proteins, namely, the 27 kDa and the 43 kDa. The lowest detection range for the 27 kDa gene was 3 pg. The amplification for both genes was positive only with DNA from P. brasiliensis; additionally, the mRNA for the 27 kDa gene was present only in P. brasiliensis, as indicated by the Northern analysis. The standardization of PCR technology permitted the amplification of P. brasiliensis DNA in artificially contaminated soils and in tissues of armadillos naturally infected with the fungus. These results indicate that PCR technology could play an important role in the search for P. brasiliensis’ habitat and could also be used in other ecological studies.

NEW PRIMERS FOR DETECTION OF Leishmania infantum USING POLYMERASE CHAIN REACTION

SUMMARY Leishmania infantum causes visceral leishmaniasis (VL) in the New World. The diagnosis of VL is confirmed by parasitological and serological tests, which are not always sensitive or specific. Our aim was to design new primers to perform a Polymerase Chain Reaction (PCR) for detecting L. infantum. Sequences of the minicircle kinetoplast DNA (kDNA) were obtained from GenBank, and the FLC2/RLC2 primers were designed. Samples of DNA from L. infantum, Leishmania amazonensis, Leishmania braziliensis, Leishmania guyanensis, Leishmania naiffi, Leishmania lainsoni, Leishmania panamensis, Leishmania major and Trypanosoma cruzi were used to standardize the PCR. PCR with FLC2/RLC2 primers amplified a fragment of 230 bp and the detection limit was 0.2 fg of L. infantum DNA. Of the parasite species assayed, only L. infantum DNA was amplified. After sequencing, the fragment was aligned to GenBank sequences, and showed (99%) homology with L. infantum. In the analysis of blood samples and lesion biopsy from a dog clinically suspected to have VL, the PCR detected DNA from L. infantum. In biopsy lesions from humans and dogs with cutaneous leishmaniasis, the PCR was negative. The PCR with FLC2/RLC2 primers showed high sensitivity and specificity, and constitutes a promising technique for the diagnosis of VL.

PERFORMANCE OF CONVENTIONAL PCRs BASED ON PRIMERS DIRECTED TO NUCLEAR AND MITOCHONDRIAL GENES FOR THE DETECTION AND IDENTIFICATION OF Leishmania spp.

In visceral leishmaniasis, the detection of the agent is of paramount importance to identify reservoirs of infection. Here, we evaluated the diagnostic attributes of PCRs based on primers directed to cytochrome-B (cytB), cytochrome-oxidase-subunit II (coxII), cytochrome-C (cytC), and the minicircle-kDNA. Although PCRs directed to cytB, coxII, cytC were able to detect different species of Leishmania, and the nucleotide sequence of their amplicons allowed the unequivocal differentiation of species, the analytical and diagnostic sensitivity of these PCRs were much lower than the analytical and diagnostic sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs had spleen and bone marrow samples collected and tested; only two animals were positive by PCRs based on cytB, coxII, and cytC, whereas 18 were positive by the kDNA-PCR. Considering the kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs had positive bone marrow results but negative results in spleen samples and, in four dogs, the reverse situation occurred. We concluded that PCRs based on cytB, coxII, and cytC can be useful tools to identify Leishmania species when used in combination with automated sequencing. The discordance between the results of the kDNA-PCR in bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity for the detection of infected dogs. Thus, primers based on the kDNA should be preferred for the screening of infected dogs.

Textos e gêneros no currículo de português da Escola Estadual Paulista

O artigo aborda os textos de trabalho incluídos nos cadernos didáticos para a disciplina de Língua Portuguesa, no projeto São Paulo Faz Escola - SPFE -, iniciado em 2008 e transformado em proposta curricular da Secretaria de Estado da Educação de São Paulo - SEE-SP. Comparam-se os usos de textos verbais e não verbais na proposta paulista e na série Português: linguagens, de William Roberto Cereja e Thereza Cochar Magalhães (Editora Saraiva), coleção didática ajustada ao Programa Nacional do Livro Didático - PNLD. O principal instrumento para análise é o conceito de gênero textual, de acordo com definição amparada pelos Parâmetros Curriculares Nacionais e pela própria SEE-SP. Entre as constatações gerais, destaca-se a secundarização da noção de gênero por parte do SPFE, em razão do uso recorrente de textos desprovidos de história social consolidada e da pasteurização dos conteúdos presentes nessa obra.

Precisão no processo de busca e recuperação da informação: uso da mineração de textos

Trata da comparação entre a indexação manual e a ferramenta de mineração de textos, por meio da análise do índice de precisão de resposta no processo de busca e recuperação da informação. O estudo de caso escolhido para o desenvolvimento da pesquisa foi o Centro de Referência e Informação em Habitação (Infohab), cuja base de dados sobre habitação, saneamento e urbanização foi indexada de forma manual por bibliotecários da Caixa Econômica Federal, com base em uma lista de palavras-chave. Houve o desenvolvimento de um protótipo cujos itens bibliográficos correspondem às teses e dissertações contidas no Infohab, o que permitiu a aplicação do software BR/Search para a execução da mineração de textos. As pesquisas no Infohab e no protótipo foram realizadas a partir da demanda de especialistas da Caixa nos assuntos contidos na base. Conclui que não há ganhos significativos na precisão ao se aplicar a ferramenta de mineração de textos em relação à indexação manual.

A pragmática no contexto da identificação de autoria de textos

Trata-se de artigo que estuda a viabilidade de identificar a autoria de textos a partir de elementos da linguística, mais precisamente da pragmática, considerando as inter-relações entre os elementos textuais e objetos externos, ligados a particularidades do "punho escritor", a exemplo dos fenômenos sociais em que o autor está inserido. A ideia é buscar a identidade textual a partir da análise do estilo, consideradas duas categorias discursivas: modalidade e avaliação.

Aplicação da descoberta de conhecimento em textos para apoio à construção de indicadores infométricos para a área de C&T

Relata resultados de pesquisa aplicando a descoberta de conhecimento em texto (DCT) em conteúdos textuais, importantes fontes de informação para tomada de decisão. O objetivo central da pesquisa foi verificar a eficácia da DCT na descoberta de informações para apoio à construção de indicadores e definição de políticas públicas. O estudo de caso foi o Serviço Brasileiro de Respostas Técnicas (SBRT) e a técnica aplicada a de agrupamento de documentos a partir dos termos minerados na base de dados. Comprovou-se a aplicabilidade da DCT na extração de informações ocultas em documentos textuais para subsidiar a tomada de decisão e a construção de indicadores, informações essas que não poderiam ser visualizadas utilizando-se recursos tradicionais de recuperação da informação. Observou-se a preocupação com o meio ambiente nas demandas feitas pelos usuários do SBRT e a aplicabilidade da DCT para orientação de políticas internas à rede SBRT.

Estabilidade relativa de alcenos: análise dos critérios encontrados nos livros textos de graduação e uma proposta de explicação operacional para alcenos dissubstituídos

Despite of being used as thermodynamic criterion to rank alkene stability in a number of undergraduate textbooks, the heat of hydrogenation does not describe adequately the relative stability of disubstituted alkenes. In this work, both the heat of formation and the heat of combustion were used as thermodynamic criteria to rank correctly the stability of alkenes according to the degree of alkyl substitution and also in the disubstituted series (geminal > trans > cis). An operational model based on molecular orbital and valence bond representations of hyperconjugation is proposed to show how this effect can explain the order of stability of this class of compounds.

Textos científicos de autoria de graduandos em química: análise dos professores

This study aimed to analyze the aspects of scientific language considered by chemistry professors in the evaluation of scientific texts authored by undergraduate chemistry students. Interviews were conducted with four professors who had evaluated reports and scientific articles written by students enrolled in undergraduate chemistry courses. The professors highlighted aspects pertaining to the structure and general organization of scientific texts typically adopted by the scientific community, and mentioned the importance of certain rhetoric elements in scientific language. This study can be an impetus for further investigation into the importance of language in chemistry education.

Telomere and microsatellite primers reveal diversity among Sclerotinia sclerotiorum isolates from Brazil

Sclerotinia sclerotiorum, the causal agent of white mold, is a problem of winter bean (Phaseolus vulgaris) production in Brazil under center-pivot irrigation. Isolates of S. sclerotiorum were obtained from a center-pivot-irrigated field near Guaíra-SP, Brazil. Mycelial compatibility group (MCG) studies revealed the presence of only two MCG. PCR/RFLP analysis of the ITS1-5.8S-ITS2 ribosomal subunit regions of these field isolates of S. sclerotiorum failed to show any genetic differences between these two MCGs. DNA amplification with a chromosomal telomere sequence-based primer and one microsatellite primer revealed genetic polymorphisms among isolates within the same MCG. Isolates taken from beans and two other crops from another region of Brazil showed the same two MCG and had identical banding patterns for the telomere and microsatellite primers. These findings support the use of telomere sequence-based primers for revealing genotypic differences among S. sclerotiorum isolates.