Exposure to Mycobacterium avium Decreases the Protective Effect of the DNA Vaccine pVAXhsp65 Against Mycobacterium tuberculosis-Induced Inflammation of the Pulmonary Parenchyma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immunosuppression in paracoccidioidomycosis: T cell hyporesponsiveness to two Paracoccidioides brasiliensis glycoproteins that elicit strong humoral immune response

To assess human cellular immune response to paracoccidioidomycosis (PCM), lymphocyte proliferative responses to purified antigens from Paracoccidioides brasiliensis were determined in healthy persons previously infected by the fungus (positive donors), in healthy noninfected persons (controls), and in PCM patients. Affinity-purified gp70 and gp43, the two major antigens in humoral immune responses, were used, Both induced lymphocyte proliferation (gp43 species-specific) in positive donors but not in controls; healthy persons previously infected by Histoplasma capsulatum reacted to gp70 and not to gp43, A similar cross-reactivity in antibody response to gp70 was previously reported; however, antibody response to gp43 has been considered specific, Lymphocytes from PCM patients, who, unlike positive donors, have high levels of anti-gp43 and anti-gp70 antibodies, proliferated poorly with gp70 and gp43 but better with other stimuli, This dichotomy between humoral and cellular antigen-specific responses suggests a Th2 immune response in PCM, which may be related to failure to control the infection.

DELAYED-TYPE HYPERSENSITIVITY RESPONSE IN AN ISOGENIC MURINE MODEL OF PARACOCCIDIOIDOMYCOSIS

The specific delayed-type hypersensitivity (DTH) response was evaluated in resistant (A/SN) and susceptible (B10.A) mice intraperitoneally infected with yeasts from a virulent (Pb18) or from a non-virulent (Pb265) Paracoccidioides brasiliensis isolates. Both strains of mice were footpad challenged with homologous antigens. Pb18 infected A/SN mice developed an evident and persistent DTH response late in the course of the disease (90th day on) whereas B10.A animals mounted a discrete and ephemeral DTH response at the 14th day post-infection. A/SN mice infected with Pb265 developed cellular immune responses whereas B10.A mice were almost always anergic. Histological analysis of the footpads of infected mice at 48 hours after challenge showed a mixed infiltrate consisting of predominantly mononuclear cells. Previous infection of resistant and susceptible mice with Pb18 did not alter their DTH responses against heterologous unrelated antigens (sheep red blood cells and dinitrofluorobenzene) indicating that the observed cellular anergy was antigen-specific. When fungal related antigens (candidin and histoplasmin) were tested in resistant mice, absence of cross-reactivity was noted. Thus, specific DTH responses against P. brasiliensis depend on both the host's genetically determined resistance and the virulence of the fungal isolate.

NtWBC1, an ABC transporter gene specifically expressed in tobacco reproductive organs

To identify genes specifically or predominantly expressed in the stigmas/styles and to establish their possible function in the reproductive process of plants, a tobacco stigma/style cDNA library was constructed and differentially screened, resulting in the isolation of several cDNA clones. The molecular characterization of one of these clones is described here. After sequencing the cDNA and the isolated genomic clone, it was determined that the corresponding gene encodes a protein containing an ATP-binding cassette, characteristic of ABC transporters. This gene, designated as NtWBC1 (Nicotiana tabacum ABC transporter of the White-Brown Complex subfamily), encodes a protein that contains the typical structure of the 'half-transporters' of the White subfamily. To establish the spatial expression pattern of the NtWBC1 gene, northern blot and real-time RT-PCR analyses with total RNA from roots, stems, leaves, sepals, petals, stamens, stigmas/styles, ovaries, and seeds were performed. The result revealed a transcript of 2.5 kb present at high levels in stigmas and styles and a smaller transcript (2.3 kb) present at a lower level in stamens. NtWBC1 expression is developmentally regulated in stigmas/styles, with mRNA accumulation increasing toward anthesis. In situ hybridization experiments demonstrated that NtWBC1 is expressed in the stigmatic secretory zone and in anthers, at the stomium region and at the vascular bundle. NtWBC1 is the first ABC transporter gene with specific expression in plant reproductive organs to be identified and its expression pattern suggests important role(s) in the reproductive process.

IgG subclass profile of serum antibodies to Leishmania chagasi in naturally infected and vaccinated dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Differentially transcribed genes in skeletal muscle of lambs

The objective of this study was to compare gene transcription profiles in Longissimus dorsi muscle of the following four hair sheep genetic groups, Morada Nova (MO), Brazilian Somali (SO), Santa Inĉs (SI) and 1/2 Dorper×1/2 Morada Nova (F1). These groups all display different postnatal muscle growth. The transcriptomes of the skeletal muscle of the lambs (at 200 days of age) were profiled by using oligonucleotide microarrays and reverse transcription-quantitative real-time PCR (RT-qPCR). The microarray experiment identified 262 transcripts that were differentially expressed when transcription levels were compared between the different breeds. A total of 23 transcripts among which those involved in skeletal muscle development (MyoD1 and IGFBP4), lipogenesis and adipogenesis (C/EBPδ, PPARγ and PGDS) were differentially expressed in at least in one comparison. Clustering analysis showed that there is greater similarity in gene expression between the MO and SI breeds and between F1 and SO genetic groups. The SO breed has the most distinct expression pattern. The RT-qPCR results confirmed the findings from the microarray study. A positive correlation was observed between the expression of MyoD1 and the cold carcass yield. The negative correlations between the weight and yield of cold carcass with the expression of C/EBPδ mean that the selection for adipogenesis could lead to a lower carcass weight. The GLUT3 and PYGL gene transcripts were negatively correlated with fat thickness, but ATP5G1 was positively correlated with this trait. Interestingly, many genes negatively correlated with PUFA were positively correlated with cold carcass yield. In conclusion, the present work demonstrated that there are breed-specific expression patterns in Brazilian hair sheep genetic groups. The differences in gene expression among genetic groups were consistent with their phenotypic differences. The positive correlation of the MyoD1 expression with the cold carcass yield suggests that this gene is important for tissue growth in sheep. The positive correlation of the C/EBPδ expression with PUFA provides an opportunity to select for lipid deposition in meat animals. © 2012 Elsevier B.V.

Identification of phospholipase C zeta in normospermic and teratospermic domestic cat sperm

In mammalian species, oocyte activation is initiated by oscillations in the intracellular concentration of free calcium ([Ca2+]i), which are also essential to allow embryonic development. To date, evidence supporting the hypothesis that a sperm factor is responsible for initiating oocyte activation has been presented in various mammalian species. Among the possible candidates to be the active sperm factor is the novel sperm-specific phospholipase C ζ (PLCζ), which besides its testis-specific expression is capable of initiating [Ca2+]i oscillations. In this study, we investigated the presence of PLCζ in the sperm of the domestic cat and whether normospermic and teratospermic cats differ in their PLCζ expression. Immunoblotting with anti-PLCζ antibodies confirmed the presence of an immunoreactive band of ~70 kDa in whole sperm lysates of domestic cat as well as in both soluble and insoluble fractions from this sperm. Additional immunoreactive bands, probably C- and N-terminal truncated versions of PLCζ, were also visualized in the soluble sperm fractions. Interestingly, immunoreactivity of PLCζ was detectable in teratospermic sperm, although with slightly less intensity than in normospermic sperm. In conclusion, domestic cat sperm express PLCζ in both cytosolic and high-pH fractions, which is consistent with data in other mammals. Sperm from teratospermic cats also express PLCζ, albeit at reduced concentrations, which may affect the fertility of these males. © 2013 Elsevier Inc..

Identificação e caracterização de promotores de genes de café (coffea arabica)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização de promotores de Eucalipto com expressão tecido-específica: raiz e folha

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolamento e caracterização de promotores órgão-específicos a partir de informações do Banco FORESTs (Eucalyptus Genome Sequencing Project Consortium)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos da clonagem na inativação do cromossomo X e da regulação nutricional do metabolismo da metionina na qualidade ovocitária em bovinos

Pós-graduação em Medicina Veterinária - FMVZ

Identicação e caracterização de dois promotores de eucalipto: com padrão de expressão ubíquo e específico de câmbio

Pós-graduação em Ciências Biológicas (Genética) - IBB

Mycetoma caused by Nocardia brasiliensis in an immunocompetent patient

Dendritic cells play a central role in the regulation of immunological reactivity. The existence of functionally specialized populations of dendritic cells in the skin is a consequence of qualitatively different attacks on our organism. slanDCs are human inflammatory dendritic cells that are characterized by the specific expression of the carbohydrate 6-sulfo LacNAc (slan). After phenotypic maturation, slanDCs are capable of producing very high amounts of proinflammatory mediators such as IL-12, TNF-, IL-1 and IL-23. Recent data describe a potential role of slanDCs in a number of different diseases like psoriasis, lupus erythematosus, and tumors, thus opening up new areas of research on their respective pathogenesis. Furthermore, a slanDC-specific targeting system has been developed as a basis for direct therapeutic manipulation. Future challenges of slanDC research include deepening our understanding of the significance of slanDCs in the regulation of adaptive and innate immune responses, as well as translating this knowledge into therapeutic options.

Amplificação, clonagem e sequenciamento de promotores de café (Coffea arabica cv Mundo Novo) específicos de fruto e ubíquos

The Coffee Genome Project made available to the scientific community relevant information that made practical the identification and cloning of important genes, as well as the identification of the major sequences involved on their regulation. The aim of the present study was to amplify, clone and sequence coffee promoters with specific expression patterns. For that, coffee ESTs which known expression profiles were employed. First, the promoter regions of coffee genes showing, respectively, fruitspecific and ubiquitous expression were amplified using the Genome Walking strategy. Amplified sequences were then inserted in the pGEM-Teasy vector (Promega) and sequenced. Once completed the sequencing, an expression cassette was constructed using the binary vector pCAMBIA-1381z (Cambia). These expression cassettes were cloned into Agrobacterium tumefaciens, and transgenic tobacco plants were generated aiming the functional characterization of these promoters

Investigação de elementos de transposição em populações de Drosophila mojavensis e D. arizonae e seus híbridos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)