INGAP-PP up-regulates the expression of genes and proteins related to K-ATP(+) channels and ameliorates Ca2+ handling in cultured adult rat islets

Islet Neogenesis Associated Protein (INGAP) increases pancreatic beta-cell mass and potentiates glucose-induced insulin secretion. Here, we investigated the effects of the pentadecapeptide INGAP-PP in adult cultured rat islets upon the expression of proteins constitutive of the K-ATP(+) channel, Ca2+ handling, and insulin secretion. The islets were cultured in RPMI medium with or without INGAP-PP for four days. Thereafter, gene (RT-PCR) and protein expression (Western blotting) of Foxa2, SUR1 and Kir6.2, cytoplasmic Ca2+ ([Ca2+](i)), static and dynamic insulin secretion, and Rb-86 efflux were measured. INGAP-PP increased the expression levels of Kir6.2, SUR1 and Foxa2 genes, and SUR1 and Foxa2 proteins. INGAP-PP cultured islets released significantly more insulin in response to 40 mM KCl and 100 mu M tolbutamide. INGAP-PP shifted to the left the dose-response curve of insulin secretion to increasing concentrations of glucose (EC50 of 10.0 +/- 0.4 vs. 13.7 +/- 1.5 mM glucose of the controls). It also increased the first phase of insulin secretion elicited by either 22.2 mM glucose or 100 mu M tolbutamide and accelerated the velocity of glucose-induced reduction of Rb-86 efflux in perifused islets. These effects were accompanied by a significant increase in [Ca2+](i) and the maintenance of a considerable degree of [Ca2+](i) oscillations. These results confirm that the enhancing effect of INGAP-PP upon insulin release, elicited by different secretagogues, is due to an improvement of the secretory function in cultured islets. Such improvement is due, at least partly, to an increased K-ATP(+) channel protein expression and/or changing in the kinetic properties of these channels and augmented [Ca2+](i) response. Accordingly, INGAP-PP could potentially be used to maintain the functional integrity of cultured islets and eventually, for the prevention and treatment of diabetes. (C) 2008 Elsevier B.V. All rights reserved.

The Effects of N Helix Deletion and Mutant F29W on the Ca2+ Binding and Functional Properties of Chicken Skeletal Muscle Troponin C

To assess the structural and functional significance of the N helix (residues 3-13) of avian recombinant troponin C (rTnC), we have constructed NHdel, in which residues 1-11 have been deleted, both in rTnC and in the spectral probe mutant F29W (Pearlstone, J. R., Borgford, T., Chandra, M., Oikawa, K., Kay, C. M., Herzberg, O., Moult, J., Herklotz, A., Reinach, F. C., and Smillie, L.B. (1992) Biochemistry 31, 6545-6553). Comparison of the far- and near-UV CD spectra (±Ca2+) of F29W and F29W/ NHdel and titration of the Ca2+-induced ellipticity and fluorescence changes indicates that the deletion has little effect on the global fold of the molecule but reduces the Ca2+ affinity of the N domain, but not the C domain, by 1.6-1.8-fold. Comparisons of the mutants NHdel, F29W, and F29W/NHdel with rTnC have been made using several functional assays. In reconstituted troponin-tropomyosin actomyosin subfragment 1 and myofibrillar ATPase systems, both F29W and NHdel have significantly reduced Ca2+-activated enzymic activities. These effects are cumulative in the double mutant F29W/ NHdel. On the other hand, maximal isometric tension development in Ca2+-activated reconstituted skinned fibers is not affected with F29W and NHdel, although the Ca2+ sensitivity of NHdel in this system is markedly reduced. We conclude that both mutations, NHdel and F29W, are functionally deleterious, possibly affecting interactions of the N domain with troponin I and/or T.

Defining functional urban regions in Bahia, Brazil, using roadway coverage and population density variables

The concept of Functional Urban Regions (FURs), also called Metropolitan Regions (MRs), is not simple. It is clear, though, that they are not simply a combination of adjacent municipalities or areas. Different methods can be used for their definition. However, especially in developing countries, the application of some methods is not possible, due to the unavailability of detailed data. Alternative approaches have been developed based on spatial analysis methods and using variables extracted from available data. The objective of this study is to compare the results of two spatial analysis methods exploring two variables: population density and an indicator of transport infrastructure supply. The first method regards Exploratory Spatial Data Analyses tools, which define uniform regions based on specific variables. The second method used the same variables and the spatial analysis technique available in the computer program SKATER - Spatial 'K'luster Analysis by Tree Edge Removal. Assuming that those classifications of regions with similar characteristics can be used for identifying potential FURS, the results of all analyses were compared with one another and with the 'official' MR. A combined approach was also considered for comparison, but none of the results match the existing MR boundaries, what challenges the official definitions. (C) 2014 Elsevier Ltd. All rights reserved.

Enzymatic production of functional oligosaccharides - A review

Interest in oligosaccharide production and its general characteristics is growing. The physiological effects resulting from its ingestion make them more attractive than its sweetness, and the versatility of these carbohydrates allows their use for human and animal nutrition, pharmacology and the cosmetics industry, among others. Several microorganisms are involved in enzyme production to create oligomers with biological activity, including fructooligosaccharides, galactooligosaccharides and aminoglucanoligosaccharides. Some oligomers are currently on the market, but the search for new microorganisms producing enzymes, high-yield processes for obtaining oligosaccharides, different physiological effects, and a correlation between chemical structure and function continues.

Heterologous expression and biochemical and functional characterization of a recombinant alpha-type myotoxin inhibitor from Bothrops alternatus snake

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

Ecological strategies of Al-accumulating and non-accumulating functional groups from the cerrado sensu stricto

Coordenação de Aperfei çoamento de Pessoal de Nível Superior (CAPES)