Neonatal anoxia in rats: hippocampal cellular and subcellular changes related to cell death and spatial memory

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

Subcellular localization and kinetic characterization of a gill (Na +, K+)-ATPase from the giant freshwater prawn macrobrachium rosenbergii

The stimulation by Mg2+, Na+, K+, NH 4 +, and ATP of (Na+, K+)-ATPase activity in a gill microsomal fraction from the freshwater prawn Macrobrachium rosenbergii was examined. Immunofluorescence labeling revealed that the (Na +, K+)-ATPase α-subunit is distributed predominantly within the intralamellar septum, while Western blotting revealed a single α-subunit isoform of about 108 kDa M r. Under saturating Mg2+, Na+, and K+ concentrations, the enzyme hydrolyzed ATP, obeying cooperative kinetics with V M = 115.0 ± 2.3 U mg-1, K 0.5 = 0.10 ± 0.01 mmol L-1. Stimulation by Na+ (V M = 110.0 ± 3.3 U mg-1, K 0.5 = 1.30 ± 0.03 mmol L -1), Mg2+ (V M = 115.0 ± 4.6 U mg -1, K 0.5 = 0.96 ± 0.03 mmol L-1), NH4 + (V M = 141.0 ± 5.6 U mg -1, K 0.5 = 1.90 ± 0.04 mmol L-1), and K+ (V M = 120.0 ± 2.4 U mg-1, K M = 2.74 ± 0.08 mmol L-1) followed single saturation curves and, except for K+, exhibited site-site interaction kinetics. Ouabain inhibited ATPase activity by around 73 % with K I = 12.4 ± 1.3 mol L-1. Complementary inhibition studies suggest the presence of F0F1-, Na+-, or K +-ATPases, but not V(H+)- or Ca2+-ATPases, in the gill microsomal preparation. K+ and NH4 + synergistically stimulated enzyme activity (≈25 %), suggesting that these ions bind to different sites on the molecule. We propose a mechanism for the stimulation by both NH4 +, and K+ of the gill enzyme. © 2013 Springer Science+Business Media New York.

Efeito das isoflavonas da soja daidzeína e genisteína em células MCF-7, HB4 e OVCAR-3: estudo da citotoxicidade, indução de apoptose, cinética de proliferação celular e expressão gênica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência de genótipos de couve (Brassica oleracea L. var. acephala DC.) na biologia de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae)

Objetivou-se, com este trabalho, avaliar a influência de alguns genótipos de couve (Brassica oleracea L. var. acephala DC.) no desenvolvimento de Plutella xylostella (L., 1758) (Lepidoptera: Plutellidae). Os genótipos avaliados foram: Manteiga de Ribeirão Pires I-2620, Roxa I-919, Manteiga de São José, Manteiga de Monte Alegre, Pires 2 de Campinas, Couve Comum, Couve de Arthur Nogueira 2, Couve de Arthur Nogueira 1. Lagartas recém-eclodidas foram mantida em discos foliares de 8 cm de diâmetro para cada genótipo. Foram analisados os seguintes parâmetros: duração e viabilidade das fases larval e pupal, longevidade e fecundidade de adultos, utilizando análises paramétricas e de agrupamentos para interpretação dos dados. Observou-se um prolongamento em dias no ciclo de P. xylostella, aumento no peso de pupa e maiores valores de viabilidade e fecundidade, durante a segunda geração. O genótipo Couve de Arthur Nogueira 2 foi menos favorável ao desenvolvimento de P. xylostella nas duas gerações, e Couve Comum demonstrou maior influência negativa ao inseto na segunda geração. Manteiga de Ribeirão Pires I-2620 foi o mais suscetível nas duas gerações, agrupando com este na segunda geração Pires 2 de Campinas e Manteiga de São José.

Comparative transcriptome analysis of Paracoccidioides brasiliensis during in vitro adhesion to type I collagen and fibronectin: identification of potential adhesins

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular characterization and phylogenetic analysis of JussuMP-I: A RGD-P-III class hemorrhagic metalloprotease from Bothrops jararacussu snake venom

Snake venom metalloproteases (SVMPs) embody zinc-dependent multidomain enzymes responsible for a relevant pathophysiology in envenomation. including local and systemic hemorrhage. The molecular features responsible for hemorrhagic potency of SVMPs have been associated with their multidomains structures which can target these proteins them to several receptors of different tissues and cellular types. BjussuMP-I. a SVMP isolated from the Bothrops jararacussu venom, has been characterized as a P-III hemorrhagic metalloprotease. The complete cDNA sequence of BjussuMP-I with 1641bp encodes open reading frames of 547 amino acid residues, which conserve the common domains of P-III high molecular weight hemorrhagic metalloproteases: (i) pre-pro-peptide, (ii) metalloprotease, (iii) disintegrin-like and (iv) rich cysteine domain. BjussuMP-I induced lyses in fibrin clots and inhibited collagen- and ADP-induced platelet aggregation. We are reporting, for the first time, the primary structure of an RGD-P-III class snake venom metalloprotease. A phylogenetic analysis of the BjussuMP-1 metalloprotease/catalytic domain was performed to get new insights into the molecular evolution of the metalloproteases. A theoretical molecular model of this domain was built through folding recognition (threading) techniques and refined by molecular dynamics simulation. Then, the final BjussuMP-I catalytic domain model was compared to other SVMPs and Reprolysin family proteins in order to identify eventual structural differences, which could help to understand the biochemical activities of these enzymes. The presence of large hydrophobic areas and some conserved surface charge-positive residues were identified as important features of the SVMPs and other metalloproteases. (C) 2006 Elsevier B.V. All rights reserved.

Comparison between apo and complexed structures of bothropstoxin-I reveals the role of Lys122 and Ca2+-binding loop region for the catalytically inactive Lys49-PLA(2)s

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

Towards the prediction of essential genes by integration of network topology, cellular localization and biological process information

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

A machine learning approach for genome-wide prediction of morbid and druggable human genes based on systems-level data

Background: The genome-wide identification of both morbid genes, i.e., those genes whose mutations cause hereditary human diseases, and druggable genes, i.e., genes coding for proteins whose modulation by small molecules elicits phenotypic effects, requires experimental approaches that are time-consuming and laborious. Thus, a computational approach which could accurately predict such genes on a genome-wide scale would be invaluable for accelerating the pace of discovery of causal relationships between genes and diseases as well as the determination of druggability of gene products.Results: In this paper we propose a machine learning-based computational approach to predict morbid and druggable genes on a genome-wide scale. For this purpose, we constructed a decision tree-based meta-classifier and trained it on datasets containing, for each morbid and druggable gene, network topological features, tissue expression profile and subcellular localization data as learning attributes. This meta-classifier correctly recovered 65% of known morbid genes with a precision of 66% and correctly recovered 78% of known druggable genes with a precision of 75%. It was than used to assign morbidity and druggability scores to genes not known to be morbid and druggable and we showed a good match between these scores and literature data. Finally, we generated decision trees by training the J48 algorithm on the morbidity and druggability datasets to discover cellular rules for morbidity and druggability and, among the rules, we found that the number of regulating transcription factors and plasma membrane localization are the most important factors to morbidity and druggability, respectively.Conclusions: We were able to demonstrate that network topological features along with tissue expression profile and subcellular localization can reliably predict human morbid and druggable genes on a genome-wide scale. Moreover, by constructing decision trees based on these data, we could discover cellular rules governing morbidity and druggability.

Biologia populacional de Cyphocarax modestus (Osteichthyes, Curimatidae) no córrego Ribeirão Claro, município de Rio Claro (SP)

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

Biologia populacional de Cyphocarax modestus no córrego Ribeirão Claro, município de Rio Claro (SP)

This work aimed a better understanding of the annual cycle of Cyphocarax modestus in the reservoir of water captation of Ribeirao Claro stream. The growth parameters were estimated through the analysis of length distribution and the relationships among time of smaller growth, alimentary activity, fat accumulation and reproductive period were considered. Besides, the instantaneous rate of natural mortality was calculated. Monthly samplings were accomplished in the Ribeirao Claro stream, in the reservoir of water captation of Rio Claro city. For that, 50 m of wait net was used, with meshes of 1.5, 2.0, 2.5, 3.0 and 3.5 cm measured between adjacent knots. The ELEFAN I program was used to estimate the growth parameters, and its application was done using the FISAT program. It was also used the seasonal version of von Bertalanffy's growth curve. It was considered that the reproduction of C. modestus is annual and concentrated from December to February, allowing the identification of different modas in the distributions, an essential condition for the conduction of that analysis type. The estimated parameters were: K = 0.34/year, L [infinity] = 15.40 cm, C = 0.2, Wp = 0.6 and M = 0.949/year, with the identification of four cohorts. The physiologic sequence in the annual cycle of the specie could be noted when data of accumulated fat in the visceral cavity, alimentary activity, reproduction time and time of smaller growth were analyzed together. It was noted that with the beginning of the maturation of gonads, the energy resources stopped being invested in the growth and passed to be used for reproduction.

Validation of a mutant of the pore-forming toxin sticholysin-I for the construction of proteinase-activated immunotoxins

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

Biologia populacional de Astyanax altiparanae Garutti & Britski (Teleostei, Characidae) do médio Rio Paranapanema, Paraná, Brasil

O objetivo desse trabalho foi a determinação da estrutura populacional da espécie. Foram escolhidos quatro locais de amostragem com características ambientais distintas na bacia do rio Tibagi. As amostragens foram realizadas mensalmente no período de janeiro de 1997 a fevereiro de 1998. Um total de 1553 indivíduos foram capturados, apresentando constância em todos os trechos, com as maiores abundâncias nos trechos do rio Congonhas (76 %). Variações ocorreram entre os trechos por fase de desenvolvimento e classes de comprimento e na distribuição espacial, demonstrando que a espécie apresenta estrutura populacional diferenciada. A espécie apresentou grande capacidade adaptativa exploratória, utilizando estratégias diferenciadas na estrutura da população. A espécie apresentou uma forte tendência ao comportamento r estrategista. O melhor desenvolvimento foi identificado em Congonhas I, talvez devido a existência de condições ambientais favoráveis a manutenção da espécie.

Histological characterization of cellular types during Scinax fuscovarius oogenesis (Lutz) (Anura, Hylidae)

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

Biologia de Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae) em limão siciliano (Citrus limon Burm)

No estudo da biologia de Polyphagotarsonemus latus em limão Siciliano, foram utilizados potes plásticos circulares com capacidade de 250 ml, contendo areia esterilizada como suporte para dois frutos novos com aproximadamente 2,0 cm de diâmetro. O ensaio foi conduzido a 27,1 ± 0,5°C, umidade relativa de 67,6 ± 1,3% e fotofase contínua. O período de ovo a adulto durou 3,7 ± 0,1 dias para fêmeas e 3,6 ± 0,1 dias para machos, com sobrevivência de 100%. Após um período de pré-oviposição de 1,0 ± 0,2 dias, as fêmeas depositaram 5,6 ± 0,5 ovos por dia durante 10,5 ± 0,9 dias, totalizando 58,9 ± 6,7 ovos por fêmea. A longevidade foi de 13,4 ± 1,0 dias para fêmeas e 12,0 ± 2,4 dias para machos. A razão intrínseca de aumento (rm) foi de 0,359, a razão finita de aumento (l) de 1,43 indivíduos por fêmea por dia, o tempo médio de uma geração (T) de 10,34 dias e a taxa líquida de reprodução (Ro) de 41,0.