Influence of moderate physical training on the GH/IGF-1 axis in diabetic rats

The influence of moderate physical training on serum growth hormone (GH), insulin-like growth factor -1 (IGF-1) and binding protein ( IGFBP-3) in experimental diabetic rats was investigated. Male Wistar rats were divided into 4 groups, sedentary control (SC), trained control (TC), sedentary diabetic (SD) and trained diabetic (TD). Experimental diabetes was induced of Alloxan (35mg/b.w.) the training program consisted by swimming 5 days/week, 1 h/day, supporting a load of 2.5% b.w., during 6 weeks. Then, the rats were sacrificed and blood was collected for determinations of serum glucose, insulin, GH, IGF-1 and IGFBP-3. Samples of liver were used to evaluate glycogen, protein and DNA contents. The results were analyzed by ANOVA, and Bonferroni test and the significance level was set at 2.5%. Diabetes decreased serum GH, IGF-1, IGFBP-3 and liver glycogen stores in SD group. Physical training promoted increase in serum IGF-1 in both TC and TD groups (SC=82 +/- 15; TC= 1 03 +/- 13; SD=77 +/- 16; TD= 112 +/- 29 ng/ml) and liver glycogen store in TD group when compared to SD (SC=5.2 +/- 1.2; TC= 6.2 +/- 1; SD=2 +/- 0.5; TD=5 +/- 1.8 mg/100mg). Therefore, physical training contributes to the increase in liver glycogen content and to rise of insulin-like growth factor level in diabetic rats. It was concluded that moderate physical training promotes important adaptations related to GH-IGF-1 axis in diabetic organisms.

Endurance training improves responsiveness to insulin and modulates insulin signal transduction through the phosphatidylinositol 3-kinase/Akt-1 pathway

Background: Endurance training increases insulin-stimulated muscle glucose transport and leads to improved metabolic control in diabetic patients.Objective: To analyze the effects of endurance training on the early steps of insulin action in muscle of rats. Design: Male rats submitted to daily swimming for 6 weeks were compared with sedentary controls. At the end of the training period, anesthetized animals received an intravenous (i.v.) injection of insulin and had a fragment of their gastrocnemius muscle excised for the experiments.Methods: Associations between insulin receptor, insulin receptor substrates (IRS)-1 and -2 and phosphatidylinositol 3-kinase (PI3-kinase) were analyzed by immunoprecipitation and immunoblotting. Akt-1 serine phosphorylation and specific protein quantification were detected by immunoblotting of total extracts, and IRS-1/IRS-2-associated PI3-kinase activity were determined by thin-layer chromatography.Results: Insulin-induced phosphorylation of IRS-1 and IRS-2 increased respectively by 1.8-fold (P < 0.05) and 1.5-fold (P < 0.05), whereas their association with PI3-kinase increased by 2.3-fold (P < 0.05) and 1.9-fold (P < 0.05) in trained rats as compared with sedentary controls, respectively. The activity of PI3-kinase associated with IRS-1 and IRS-2 increased by 1.8-fold (P < 0.05) and 1.7-fold (P < 0.05) respectively, in trained rats as compared with their untrained counterparts. Serine phosphorylation of Akt-1/PKB increased 1.7-fold (P < 0.05) in trained rats in response to insulin. These findings were accompanied by increased responsiveness to insulin as demonstrated by a reduced area under the curve for insulin during an i.v. glucose tolerance test, by increased glucose disappearance rate during an insulin tolerance test, and by increased expression of glucose transporter-4.Conclusions: the increased responsiveness to insulin induced by chronic exercise in rat skeletal muscle may result, at least in part, from the modulation of the insulin signaling pathway at different molecular levels.

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.

Aggregation behavior of hydrophobically modified dextran in aqueous solution: a fluorescence probe study

Dextrans (M-W = 11.000 and M-w = 40.000) have been modified with 4-hexyl benzoyl chloride and their aggregation behavior was studied in aqueous solution employing the fluorescent probes pyrene and 1,8 anilinonaphtalene sulfonic acid sodium salt (1,8 ANS). The photophysical studies showed that above a critical concentration the derivatives tend to form aggregates having different properties, which depend on both the degree of substitution (alpha) and the molecular weight of the sample. The parameter alpha has a marked effect on the critical aggregation concentrations (CAC) and aggregate proper-ties. Hydrophobic microenvironments can be detected for substituted dextrans having alpha values varying from 0.01 to 0.19. CAC values decreased by two orders and magnitude when the molecular weight increased from 11 to 40 kDa, leading to formation of more apolar aggregates and diminishing by about 30% the polarity of the microenviromnents. Pre-aggregation was evidenced by pyrene excimer emission and intermolecular interactions were responsible by the formation of aggregates leading to solution behaviour similar to that of common surfactants. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Measurement of inclusive differential cross sections for Upsilon(1S) production in p(p)over-bar collisions at root s=1.96 TeV

We present measurements of the inclusive production cross sections of the Upsilon(1S) bottomonium state in p (p) over bar collisions at root s=1.96 TeV. Using the Upsilon(1S)->mu(+)mu(-) decay mode for a data sample of 159 +/- 10 pb(-1) collected by the D0 detector at the Fermilab Tevatron collider, we determine the differential cross sections as a function of the Upsilon(1S) transverse momentum for three ranges of the Upsilon(1S) rapidity: 0 <\y(Upsilon)\<= 0.6, 0.6 <\y(Upsilon)\<= 1.2, and 1.2 <\y(Upsilon)\<= 1.8.

Search for first-generation scalar leptoquarks in p(p)over-bar collisions at root s=1.96 TeV

We report on a search for pair production of first-generation scalar leptoquarks (LQ) in p (p) over bar collisions at root s=1.96 TeV using an integrated luminosity of 252 pb(-1) collected at the Fermilab Tevatron collider by the D0 detector. We observe no evidence for LQ production in the topologies arising from LQ(LQ) over bar -> eqeq and LQ(LQ) over bar -> eq nu q, and derive 95% C.L. lower limits on the LQ mass as a function of beta, where beta is the branching fraction for LQ -> eq. The limits are 241 and 218 GeV/c(2) for beta=1 and 0.5, respectively. These results are combined with those obtained by D0 at root s=1.8 TeV, which increases these LQ mass limits to 256 and 234 GeV/c(2).

Search for a heavy resonance decaying into a Z plus jet final state in p(p)over-bar collisions at root s=1.96 TeV using the D0 detector

We have searched for a heavy resonance decaying into a Z+jet final state in p (p) over bar collisions at a center of mass energy of 1.96 TeV at the Fermilab Tevatron collider using the D0 detector. No indication for such a resonance was found in a data sample corresponding to an integrated luminosity of 370 pb(-1). We set upper limits on the cross section times branching fraction for heavy resonance production at the 95% C.L. as a function of the resonance mass and width. The limits are interpreted within the framework of a specific model of excited quark production.

Search for scalar top quarks in the acoplanar charm jets and missing transverse energy final state in p(p)over-bar collisions at root s=1.96 TeV

We present a search for the pair production of scalar top quarks, (t) over tilde, using 995 pb(-1) of data collected in p (p) over bar collisions with the Dempty set detector at the Fermilab Tevatron Collider at root s = 1.96 TeV. Both scalar top quarks are assumed to decay into a charm quark and a neutralino ((chi) over tilde (0)(1)), where (chi) over tilde (0)(1) is the lightest supersymmetric particle. This leads to a final state with two acoplanar charm jets and missing transverse energy. We find the yield of such events to be consistent with the standard model expectation, and exclude sets of (t) over tilde and (chi) over tilde (0)(1) masses at the 95% C.L. that substantially extend the domain excluded by previous searches. Published by Elsevier B.V.

Simultaneous Determination of Ba, Cr, Mo (Group 1), and Cu, Fe, Ni, and Pb (Group 2) in Commercial Fuel Ethanol by Graphite Furnace AAS

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

Thermal behaviour of intercalated 8-hydroxyquinoline (oxine) in montmorillonite clay

The composite montmorillonite-8-hydroxyquinoline (Swy-1-8-HQ) was prepared by two different processes and studied by using thermogravimetric analysis (TG/DTG and DSC), as well as helpful techniques as fluorescence in the UV-visible region and X-ray diffraction. The composites developed fluorescent appearance, however with quantum poor efficiency and they exhibited distinct TG and DSC thermal behavior. The fluorescence data of spectra associated to the TG/DT curves allowed to suggest that the 8-HQ was present in the composites in two different circumstances: 1 - intercalated in the interlayer spaces (Swy-1-8-HQ2), rigidly associated to the Substrate feasible as a monolayer with the aromatic rings parallel to the silica layer; and/or, 2 - adsorbed on the Surface (Swy-1-8-HQ1), either as a bilayer formation or tilting of the molecules to the silicate layer sheet. All results confirmed above are in agreement with X-ray diffraction patterns, once the interlayer space increases when 8-HQ is incorporated. The experimental results confirm the formation of the composites in agreement with the method used in the preparation.

Glucose- and insulin-induced phosphorylation of the insulin receptor and its primary substrates IRS-1 and IRS-2 in rat pancreatic islets

The presence of tyrosine-phosphorylated proteins was studied in cultured rat pancreatic islets, Immunoblotting performed with total extracts of islets cultured in the presence of 1.8 or 5.6 mM glucose revealed at least three distinct tyrosine-phosphorylated bands (25 kDa, 95 kDa and 165-185 kDa). After 12 h incubation in medium containing 1.8 mM glucose, a pulse exposition to 11 or 22 mM glucose or to 10(-7) M insulin led to a substantial increase in the phosphorylation of all three bands, with no appearance of novel bands. Immunoprecipitation with specific antibodies demonstrated that the signal detected at 95 kDa corresponds to the beta subunit of the insulin receptor (IR) while the band at 165-185 kDa corresponds to the early substrates of the insulin receptor, IRS-1 and IRS-2. Immunoprecipitation with IRS-I or IRS-2 antisera detected their association with the lipid metabolizing enzyme phosphatidylinositol 3-kinase (PI 3-kinase), Thus, this is the first demonstration that elements involved in the insulin-signalling pathway of traditional target tissues are also present in pancreatic islets and are potentially involved in auto- and paracrine-signalling in this organ.

Genetic relatedness of Plasmodium falciparum isolates and the origin of allelic diversity at the merozoite surface protein-1 (MSP-1) locus in Brazil and Vietnam

Background: Despite the extensive polymorphism at the merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum, that encodes a major repetitive malaria vaccine candidate antigen, identical and nearly identical alleles frequently occur in sympatric parasites. Here we used microsatellite haplotyping to estimate the genetic distance between isolates carrying identical and nearly identical MSP-1 alleles. Methods: We analyzed 28 isolates from hypoendemic areas in north-western Brazil, collected between 1985 and 1998, and 23 isolates obtained in mesoendemic southern Vietnam in 1996. MSP-1 alleles were characterized by combining PCR typing with allele-specific primers and partial DNA sequencing. The following single-copy microsatellite markers were typed: Polyα, TA42 (only for Brazilian samples), TA81, TA1, TA87, TA109 (only for Brazilian samples), 2490, ARAII, PfG377, PfPK2, and TA60. Results: The low pair-wise average genetic distance between microsatellite haplotypes of isolates sharing identical MSP-1 alleles indicates that epidemic propagation of discrete parasite clones originated most identical MSP-1 alleles in parasite populations from Brazil and Vietnam. At least one epidemic clone propagating in Brazil remained relatively unchanged over more than one decade. Moreover, we found no evidence that rearrangements of MSP-1 repeats, putatively created by mitotic recombination events, generated new alleles within clonal lineages of parasites in either country. Conclusion: Identical MSP-1 alleles originated from co-ancestry in both populations, whereas nearly identical MSP-1 alleles have probably appeared independently in unrelated parasite lineages.

Frequency of multiple endocrine neoplasia type 1 in a group of patients with pituitary adenoma: genetic study and familial screening

The purpose of this study it was to evaluate the frequency of Multiple Endocrine Neoplasia type 1 (MEN1) in patients with pituitary adenoma and to perform genetic analysis and familial screening of those individuals afflicted with MEN1. 144 patients with pituitary adenoma at Botucatu Medical School, UNESP-Univ Estadual Paulista, were assessed retrospectively for MEN1 during the years of 2005-2011. The patients were evaluated for the presence of primary hyperparathyroidism (PHP) and enteropancreatic tumors. Genetic analysis was performed for the individuals with clinically diagnosed MEN1. Thirteen patients met the diagnostic criteria for MEN1, but three individuals belong to the same family and they were considered as a single MEN1 event, revealing 7.7 % frequency of MEN1 in this patient group. Genetic analysis showed MEN1 mutations in four index cases: IVS4+1 G>A, IVS3-6 C>T, c.1547insC and a new D180A mutation. One patient did not agree to participate in the genetic study and another one was referred for follow up in other hospital. Only polymorphisms were found in the other individuals, one of which was novel. We identified a high frequency of MEN1 in pituitary adenoma patients. Since PHP is one of the most common MEN1 tumor and patients are mostly asymptomatic, we suggest that all pituitary adenoma patients have their calcium profile analyzed. © 2013 Springer Science+Business Media New York.

Characterization of resveratrol content in ten wild species of section Arachis, genus Arachis

Peanut is one of the few plants that synthesizes resveratrol, a phenolic compound of the stilbene class, which has been associated with reduced risk of developing chronic diseases, such as cancer, cardiovascular diseases, skin diseases, pulmonary diseases, diabetes and neurological diseases. Resveratrol was detected in different parts of the peanut plant, including roots, leaves, seeds and their derivatives. The wild species of the Arachis section are also strong candidates to synthesize resveratrol because they are phylogenetically closely related to cultivated peanut. Our objective was to characterize the resveratrol content in ten wild species of Arachis with three different genomes (A, B and K). The plant material was composed of leaves of the ten species treated (test) and not treated (control) with ultraviolet (UV) radiation. The test and control samples were extracted and the identification and quantification of resveratrol was performed using high performance liquid chromatography (HPLC). All species studied synthesized resveratrol and the concentrations ranged from 299.5 μg/g in A. kempff-mercadoi to 819.9 μg/g in A. cardenasii. DPPH antioxidant activity varied between 18.7 % for A. duranensis and 48.2 % in A. simpsonii. The results showed that wild Arachis species are a potential source of alleles for improvement of cultivated peanut, with the aim of achieving higher resveratrol content in leaves. © 2013 Springer Science+Business Media Dordrecht.

Nitensidine A, a guanidine alkaloid from Pterogyne nitens, induces osteoclastic cell death

Nitensidine A is a guanidine alkaloid isolated from Pterogyne nitens, a common plant in South America. To gain insight into the biological activity of P. nitens-produced compounds, we examined herein their biological effects on osteoclasts, multinucleated giant cells that regulate bone metabolism by resorbing bone. Among four guanidine alkaloids (i.e., galegine, nitensidine A, pterogynidine, and pterogynine), nitensidine A and pterogynine exhibited anti-osteoclastic effects at 10 μM by reducing the number of osteoclasts on the culture plate whereas galegine and pterogynidine did not. The anti-osteoclastic activities of nitensidine A and pterogynine were exerted in a concentration-dependent manner, whereas nitensidine A exhibited an approximate threefold stronger effect than pterogynine (IC50 values: nitensidine A, 0.93 ± 0.024 μM; pterogynine, 2.7 ± 0.40 μM). In the present study, the anti-osteoclastic effects of two synthetic nitensidine A derivatives (nitensidine AT and AU) were also examined to gain insight into the structural features of nitensidine A that exert an anti-osteoclastic effect. The anti-osteoclastic effect of nitensidine A was greatly reduced by substituting the imino nitrogen atom in nitensidine A with sulfur or oxygen. According to the differences in chemical structures and anti-osteoclastic effects of the four guanidine alkaloids and the two synthetic nitensidine A derivatives, it is suggested that the number, binding site, and polymerization degree of isoprenyl moiety in the guanidine alkaloids and the imino nitrogen atom cooperatively contribute to their anti-osteoclastic effects. © 2013 Springer Science+Business Media Dordrecht.