Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Ação Sacietogênica de um inibidor de tripsina da semente de tamarindo (Tamarindus indica L.)

The seeds are excellent sources of proteinase inhibitors and have been highlighted owing to various applications. Among these applications are those in effect on food intake and weight gain that stand out because of the increasing number of obese individuals. This study evaluated the effects of trypsin inhibitor present in the seed of tamarind (Tamarindus indica L.) reduction in weight gain, biochemical and morphological alterations in Wistar rats. For this, we partially purified a trypsin inhibitor tamarind seed. This inhibitor, ITT2 at a concentration of 25 mg / kg body weight, over a period of 14 days was able to reduce food intake in rats (n = 6) by approximately 47%, causing a reduction in weight gain approximately 70% when compared with the control group. With the evaluation of the in vivo digestibility was demonstrated that the animals lost weight due to satiety, presented by the reduction of food intake, since there were significant differences between true digestibility for the control group (90.7%) and the group treated with inhibitor (89.88%). Additionally, we checked the deeds of ITT2 on biochemical parameters (glucose, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, gamma glutamyl transferase albumin, globulin, total protein and C-reactive protein) and these, when assessed in the study groups showed no statistically significant variations. We also evaluate the histology of some organs, liver, stomach, intestine, and pancreas, and showed no changes. And to evaluate the effect of trypsin inhibitor on food intake due to the satiety is regulated by cholecystokinin (CCK) were measured plasma levels, and it was observed that the levels of CCK in animals receiving ITT2 were significantly higher ( 20 + 1.22) than in animals receiving only solution with casein (10.14 + 2.9) or water (5.92 + 1.15). Thus, the results indicate that the effect caused ITT2 satiety, reducing food intake, which in turn caused a reduction in weight gain in animals without causing morphological and biochemical changes, this effect caused by the elevation of plasma levels CCK

Parâmetros citogenéticos de espécies comerciais de carangidae (perciformes), com vistas a sua empregabilidade na conservação biológica e piscicultura marinha

Worldwide, families Carangidae and Rachycentridae represent one of the groups most important commercial fish, used for food, and great potential for marine aquaculture. However, the genetic bases that can underpin the future cultivation of these species, cytogenetic between these aspects are very weak. The chromosomal patterns have provided basic data for the exploration of biotechnological processes aimed at handling chromosomal genetic improvement, such as induction of polyploidy, androgenesis and ginogenesis, as well as obtaining monosex stocks and interspecific hybridizations. This paper presents a comprehensive cytogenetic survey in 10 species, seven of the family Carangidae and the monotypic family Rachycentridae. Classical cytogenetic analysis and in situ mapping of multigene sequences were employed, and additionally for the genus Selene and morphotypes of Caranx lugubris, comparisons were made using geometric morphometrics. In general, conservative species exhibit a marked chromosome number (2n=48). Although present in large part, different karyotypic form, retain many characteristics typical of chromosomal Order Perciformes, the high number of elements monobrachyal, Ag-NORs/18S rDNA sites and heterochromatin simply reduced, preferably centromeric. The main mechanisms involved in karyotypic diversification are the pericentric inversions, with secondary action of centric fusions. In addition to physical mapping and chromosome detail for the species are presented and discussed patterns of intra-and interspecific diversity, cytotaxonomic markers. This data set provides a better understanding of these patterns caryoevolutyonary groups and conditions for the development of protocols based on Biotechnology for chromosomal manipulation Atlantic these species

Diversidade de invertebrados em cavernas calcárias do oeste potiguar : subsídios para a determinação de áreas prioritárias para conservação.

It is estimated that the Brazilian karst areas sum about 200.000 km2. The caves, one of the main components of karst, are important windows into the biological studies on hypogean environments. In Rio Grande do Norte are known 563 caves, and 476 of them are in the municipalities of Baraúna, Felipe Guerra, Governador Dix-Sept Rosado, Apodi and Mossoró, the Western region of the State. However, like in the rest of the country, the cave fauna of the State is still poorly understood. This study used data from invertebrates harvested in 47 caves and aimed to analyze the effect of environmental change between the dry and rainy seasons in the communities of cave invertebrates, characterize these communities and evaluate the relationships between morphological and biotic variables of the caves and surroundings, and to define priority areas for conservation of cave environments of the study area from biotic parameters. Strong effects were found in the community structure of cave invertebrates due environmental changes between seasons, with values of total richness, abundance, diversity and ecological complexity significantly higher in the rainy season. It was possible to assess how the morphology of the cave and the external environment variables affect the biotic system, so that the variety of resources, forest cover in the vicinity, the area of the cave and its entrance were the variables that best explained the structure communities of cave invertebrates in the region. High values of total richness of invertebrates (36,62 ± 14,04 spp / cave) and troglomorphic species (61 species, mean 1,77 ± 2,34 spp / cave) were found and, given the biological relevance in the context of the area national and the imminent anthropogenic pressures existing, we defined four priority areas for actions aiming cave biodiversity conservation in the region.

Conservação de filés de tilápia-do-nilo (Oreochromis niloticus) em salga seca e salga úmida

Acompanhou-se o desenvolvimento dos processos da salga em salmoura saturada (salga úmida) e salga seca de filés de tilápia-do-nilo (Oreochromis niloticus) e avaliaram-se algumas características indicativas de qualidade do produto durante a estocagem. Os processos foram acompanhados por 156 horas na salga úmida e por 96 horas na salga seca, e os filés salgados foram estocados, respectivamente, por 60 e 45 dias à temperatura ambiente. Os teores máximos de cloreto nos filés (14%) foram atingidos com 72 horas na salga úmida e com 36 horas na salga seca. Os filés de tilápia salgados em salmoura mantiveram as características próprias do produto por um período de 45 dias, e os submetidos à salga seca apresentaram baixo teor de umidade (6%) e alta concentração de extrato etéreo (4,6%). Recomenda-se somente o processo de salga em salmoura saturada como forma de conservação dos filés de tilápia-do-nilo.