Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection

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

Molecular approaches for eco-epidemiological studies of Paracoccidioides brasiliensis

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

Inteins in pathogenic fungi: a phylogenetic tool and perspectives for therapeutic applications

Inteins or internal proteins are coding sequences that are transcribed and translated with flanking sequences (exteins). After translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. These parasitic genetic elements have been found in important, conserved proteins in all three domains of life. Most of the eukaryotic inteins are present in the fungi kingdom and the PRP8 intein is one of the most widespread inteins, occurring in important pathogens such as Cryptococcus neoformans (varieties grubii and neoformans), Cryptococcus gattii, Histoplasma capsulatum and Paracoccidioides brasiliensis. The knowledge of conserved and non-conserved domains in inteins have opened up new opportunities for the study of population variability in pathogenic fungi, including their phylogenetic relationships and recognition or diagnoses of species. Furthermore, inteins in pathogenic fungi should also be considered a promising therapeutic drug target, since once the autocatalytic splicing is inhibited, the host protein, which is typically vital, will not be able to perform its normal function and the fungal cell will not survive or reproduce.

Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping 'hot spot' areas of PCM.

PRP8 intein in Ajellomycetaceae family pathogens: Sequence analysis, splicing evaluation and homing endonuclease activity

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

Genus Paracoccidioides: Species Recognition and Biogeographic Aspects

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

Acidentes do trabalho em pescadores artesanais da região do Médio Rio Araguaia, Tocantins, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nigriventrine: A low molecular mass neuroactive compound from the venom of the spider Phoneutria nigriventer

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

Nodule ultrastructure and initial growth of Anadenanthera peregrina (L.) Speg. var. falcata (Benth.) Altschul plants infected with rhizobia

The anatomy and ultrastructure of root nodules of Anadenanthera peregrina var. falcata (Leguminosae-Mimosoideae) were analysed, as was plant growth. To ensure that nodules developed, seedlings were inoculated with a mixture of six strains of rhizobia. Nodules were produced that differed in appearance-and probably also effectiveness-but their structure was similar and they showed characteristics typical of indeterminate nodules, such as persistent meristematic tissue and a gradient of cells at different stages of development. Many starch grains were present in inner cortex cells and interstitial cells of infected tissue. Infected cells were densely packed with bacteroids, which contained many poly-beta-hydroxybutyrate granules. The high incidence of these granules, together with high levels of starch accumulation in interstitial cells, suggested low N-2-fixation efficiency of the rhizobia isolates used for inoculation. In the symbiosomes of early-senescent infected cells, reticulum-like structures, small vesicles and a fibrillar material were observed; these may be related to bacteroid degradation. In the cytoplasm of late-senescent infected cells, many vesicles and membrane-like structures were observed, probably associated with membrane degradation of bacteroids and peribacteroids. The total biomass of plants inoculated with rhizobia was low and their xylopodia and shoots had low levels of N compared with non-inoculated plants fertilized with ammonium nitrate. However, inoculated plants did not show N-deficiency symptoms and grew better than non-inoculated plants without N fertilization. These growth results, together with ultrastructural observations of nodules, suggest that nitrogen fixation of rhizobia isolates associated with Anadenanthera peregrina var. falcata roots is poor. (C) 2002 Annals of Botany Company.

Nitrogen-tixing and vesicular-arbuscular mycorrhizal symbioses in some tropical legume trees of tribe Mimoseae

Response to mineral fertilization and inoculation with rhizobia and/or arbuscular mycorrhiza fungi (AMF) of the Anadenanthera colubrina, Mimosa bimucronata and Parapiptadenia rigida (Leguminosae-Mimosoideae) native trees from Brazilian riparian forests, were studied in nursery conditions. Each species was submitted to seven treatments, varying nitrogen and phosphorous fertilization and inoculation with rhizobia (r), mycorrhiza (m) or both (rm): NP, P, P + r, P + rm, N, N + m and N + rm. Results showed that AMF inoculations did not enhance the mycorrhizal colonization, and P uptake was not sufficient to sustain good growth of plants. The level of P mineral added affected negatively the AMF colonization in A. colubrina and M. bimucronata, but not in P. rigida. Native fungi infected the three legume hosts. The absence of mineral N limited growth of A. colubrina and P. rigida, but in M. bimucronata the lack of N was corrected by biological nitrogen fixation. N mineral added inhibited the nodulation, although spontaneous nodulation had occurred in A. colubrina and M. bimucronata. Rhizobia inoculation enhanced the number of nodules, nitrogenase activity and leghemoglobin content of these two species. Thus, the extent of rhizobial and mycorrhizal symbiosis in these species under nursery conditions can affect growth and consequently the post-planting success. (C) 2004 Elsevier B.V. All rights reserved.

Nodulação e micorrização em Anadenanthera peregrina var. falcata em solo de cerrado autoclavado e não autoclavado

Anadenanthera peregrina var. falcata (angico-do-cerrado), uma leguminosa arbórea, forma associações simbióticas com bactérias fixadoras de nitrogênio (rizóbios) e com fungos micorrízicos arbusculares. Com o objetivo de avaliar a eficiência da inoculação de fungos micorrízicos e rizóbios no crescimento inicial de plantas de angico-do-cerrado, crescidas em solo autoclavado e em solo não autoclavado com e sem inoculação, foi desenvolvido um experimento em casa de vegetação, utilizando raízes micorrizadas de milho e uma mistura de isolados de rizóbios como inoculantes. O crescimento das plantas foi influenciado positivamente pela concomitante inoculação do fungo micorrízico e do rizóbio, tendo as plantas desse tratamento apresentado biomassa cerca de 60 % maior do que o controle no décimo mês. A inoculação de apenas um dos microssimbiontes, entretanto, não provocou diferença na produção de biomassa das plantas. A percentagem de colonização micorrízica foi significativamente mais alta e o número de nódulos maior nas raízes das plantas crescidas no solo não autoclavado, ocasionados pela população de fungos e rizóbios nativos. Nesse tratamento, houve pequeno acúmulo de matéria no xilopódio, provavelmente em virtude do dreno fotossintético por parte dos microssimbiontes, e a concentração de P na parte aérea e xilopódio dessas plantas foi cerca de 1,2 e 8 vezes maior, respectivamente, por causa da colonização micorrízica.

Morphology, development and homologies of the perianth and floral nectaries in Croton and Astraea (Euphorbiaceae-Malpighiales)

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

Capacidade funcional, desempenho e solicitação metabólica em futebolistas profissionais durante situação real de jogo monitorados por análise cinematográfica

Foi objetivo deste estudo caracterizar a relação entre o nível de aptidão física, desempenho e solicitação metabólica em futebolistas durante situação real de jogo. Seis jogadores de futebol profissional com média de idade de 20,8 ± 2,6 anos (17-25), peso 70,4 ± 7,5kg (63-81,3) e altura 173,3 ± 9,7cm (166-188), foram submetidos a testes de aptidão física em campo e análise cinematográfica durante a partida. Os testes de aptidão física foram realizados em campo, com medições de lactato sanguíneo. A via metabólica alática foi avaliada por meio de cinco corridas na distância de 30m, em velocidade máxima, com pausa passiva de um minuto entre cada corrida. As concentrações de lactato foram medidas no 1º, 3º e 5º minuto após o término das cinco corridas. Para detecção do limiar anaeróbio foram realizadas 3 corridas de 1.200m nas intensidades de 80, 85 e 90% da velocidade máxima para essa distância, com intervalo passivo de 15 minutos entre cada corrida. As dosagens de lactato sanguíneo foram feitas no 1º, 3º e 5º minuto de repouso passivo após cada corrida. Os futebolistas foram submetidos à filmagem individual durante o transcorrer do jogo e as concentrações de lactato foram medidas antes, no intervalo e no final da partida para análise da solicitação energética e metabólica, respectivamente. Os seguintes resultados foram verificados: 1) o limiar anaeróbio em velocidade de corrida, correspondente à concentração de lactato sanguíneo de 4mmol.L_1 foi encontrado aos 268 ± 28m.min_1 ou 16,1 ± 1,6km.h_1; 2) a velocidade média e a concentração de lactato máximo nas corridas de 30m foram de 6,9 ± 0,2m.s_1 e 4,5 ± 1,0mmol.L_1, respectivamente; 3) a distância total percorrida foi de 10.392 ± 849m, sendo 5.446 ± 550m para o primeiro e 4.945 ± 366m para o segundo tempo, respectivamente; 4) os valores médios encontrados nas concentrações de lactato sanguíneo foram de 1,58 ± 0,37; 4,5 ± 0,42 e 3,46 ± 1,54mmol.L_1 antes, no intervalo do primeiro para o segundo tempo e ao final da a,respectivamente; e 5) a distância média total atingida ao final das partidas pelos jogadores de meio-campo (10.910 ± 121m) foi ligeiramente maior que a percorrida pelos atacantes (10.377 ± 224m) e defensores (9.889 ± 102m), mas não significativa. Houve correlação negativa (r =- 0,84; p < 0,05) entre o limiar anaeróbio (268 ± 28m.min_1 ou 16,1 ± 1,6km.h_1) e a concentração de lactato sanguíneo (4,5 ± 0,4 mmol.L_1) no primeiro tempo do jogo. Portanto, os resultados sugerem que a capacidade aeróbia é um determinante importante para suportar a longa duração da partida e recuperar mais rapidamente os futebolistas dos esforços realizados em alta intensidade, com o desenvolvimento de concentrações de lactato sanguíneo menores ao final do primeiro e segundo tempo das partidas.

Methylation profile of genes CDKN2A (p14 and p16), DAPK1, CDH1, and ADAM23 in head and neck cancer

Hypermethylation in the promoter region has been associated with a loss of gene function that may give a selective advantage to neoplastic cells. In this study, the methylation pattern of genes CDKN2A (alias p14, p14(ARF), p16, p16(INK4a)), DAPK1, CDH1, and ADAM23 was analyzed in 43 samples of head and neck tumors using methylation-specific polymerase chain reaction. In the oropharynx, there was a statistically significant association between hypermethylation of the DAPK1 gene and the occurrence of lymph node metastases, and in the larynx there was statistically significant evidence of an association between hypermethylation of the ADAM23 gene and advanced stages of the tumors. Thus, a correlation was observed between hypermethylation of the promoter region of genes DAPK1 and ADAM23 and the progression of head and neck cancer. (c) 2007 Elsevier B.V. All rights reserved.

Tissue alterations in the Guinea pig lateral prostate following antiandrogen flutamide therapy

The flutamide antiandrogenic effects oil the Guinea pig male prostate morphology in puberal, post-puberal and adult ages were evaluated in the present study. Daily-treated group animals received flutamide Subcutaneous injection at a dose of 10 mg/Kg body weight for 10 days. The control group animals received a pharmacological vehicle under the same conditions. The lateral prostate was removed, fixed and processed for light and transmission electron microscopy. The results revealed all increase of the acinus diameter in the treated puberal animals and straitness in the stromal compartment around the acini. The epithelial cells exhibited cubic phenotype. In the post-puberal and adult animals, a decrease of the acinus diameter was observed, as well as an increase of the smooth muscle layer and presence of the folds at epithelium. The ultrastructural evaluation of the secretory cells in the treated group demonstrated endomembrane enlargement, mainly in the rough endoplasmic reticulum and Golgi apparatus. In addition, a decrease of the microvilli and alterations in the distribution patterns and density of the stromal fibrillar components were observed. In Conclusion, the flutamide treatment exerts tissue effects oil the lateral prostate, promoting stroma/epithelium alterations.