Mg2+-dependent ATPase activity in Malpighian tubules of Triatoma infestans Klug

Mg2+-dependent ATPases were investigated in Malpighian tubules of the blood-sucking insect, Triatoma infestans, with cytochemical procedures for light and electron microscopy. The aim was to establish patterns of enzyme occurrence in the blood-sucking insect under control rearing conditions for further comparisons with animals subjected to the action of stress factors. Enzyme activity was found in laminated "concretions" present in distal cells, in edges of urate crystals at the lumen of the proximal region of tubules, in the basement membrane of proximal cells, and variously distributed in plasmalemma invaginations of both distal and proximal cells. Presence of ATPases in the "concretions" and urate crystals is presumed to be due to engulfment of other ATPase-containing components during formation of these structures. Cytochemical reactivity in the basement membrane and plasmalemma invaginations is assumed to be involved with active transport of waste molecules from and to hemolymph and differs as a function of the Malpighian tubule region. This paper provides a basic understanding of the enzyme occurrence in the blood sucking insects, and can be used as a pattern for comparative means of the staining patterns among Triatominae species. (C) 2011 Elsevier Ltd. All rights reserved.

Protease inhibition activity of extracts from Salicaceae species from Brazilian Cerrado and Atlantic Rain Forest and of an enriched fraction of clerodane diterpenes (casearins)

Considerando o uso popular de Casearia sylvestris Sw., Salicaceae, para o tratamento de problemas gástricos e resultados pré-clínicos que mostraram potencial atividade anti-ulcerogênica, foi realizado um screening farmacológico para avaliar a atividade biológica de outras espécies de Salicaceae. Para isso, foi utilizado um ensaio de inibição de proteases como um modelo farmacológico molecular para screening de extratos com atividade anti-ulcerogênica. Os extratos etanólico e aquoso dos galhos e folhas de C. gossypiosperma, C. decandra e C. rupestris mostraram inibição da atividade da pepsina em aproximadamente 50% com a concentração de 1 μg/mL. Curiosamente, C. obliquoa e Flacourtia ramontchi não apresentaram atividade sobre a pepsina, mas seus extratos mais apolares mostraram atividade inibitória sobre a subtilisina. A fração enriquecida de diterpenos clerodânicos mostrou atividade inibitória (42,75%) sobre a pepsina com a concentração de 1 μg/mL, mas não sobre a subtilisina (23,76%). Os resultados obtidos com os extratos e folhas das espécies testadas mostraram um padrão de atividade diferente sobre os dois tipos de proteases, a pepsina e a subtilisina, as quais estão relacionadas com diferentes tipos de atividades biológicas. Ainda mais, os resultados com a fração enriquecida de diterpenos clerodânicos sugerem que estas substâncias podem estar relacionadas com a atividade do extrato bruto de C. sylvestris.

A structure-activity relationship study of quinone compounds with trypanocidal activity

A set of 25 quinone compounds with anti-trypanocidal activity was studied by using the density functional theory (DFT) method in order to calculate atomic and molecular properties to be correlated with the biological activity. The chemometric methods principal component analysis (PCA), hierarchical cluster analysis (HCA), stepwise discriminant analysis (SDA), Kth nearest neighbor (KNN) and soft independent modeling of class analogy (SIMCA) were used to obtain possible relationships between the calculated descriptors and the biological activity studied and to predict the anti-trypanocidal activity of new quinone compounds from a prediction set. Four descriptors were responsible for the separation between the active and inactive compounds: T-5 (torsion angle), QTS1 (sum of absolute values of the atomic charges), VOLS2 (volume of the substituent at region B) and HOMO-1 (energy of the molecular orbital below HOMO). These descriptors give information on the kind of interaction that occurs between the compounds and the biological receptor. The prediction study was done with a set of three new compounds by using the PCA, HCA, SDA, KNN and SIMCA methods and two of them were predicted as active against the Trypanosoma cruzi. (c) 2005 Elsevier SAS. All rights reserved.

Relationship between carbon source, production and pattern action of alpha-amilase from Rhizopus sp

This paper discusses the inducer effect of corn soluble starch and the individual components (amylose and amylopectin) from corn and potatoes starch for alpha-amylase production by a strain of Rhizopus sp. The following decreasing order in the enzyme production was obtained: corn amylose > potatoes amylose > corn amylopectin > potatoes amylopectin > starch > maltose, coinciding with the ability of the enzyme to release reducing units, except the soluble starch that was more softly hydrolysed. However, when the enzyme action was measured by the iodine binding method, an inverse order of enzyme activity was obtained, that is: amylopectins > starch > amylosis. The results suggest that: a) branched structures in substrate affect the enzyme production; b) corn amylose and corn amylopectin are better inducers than their respectives homologous from potatoes; c) cc-amylase from Rhizopus sp has different action patterns on substrates with straight or branched chains: from the former, it removes only reducing units with lower molecular weight (G1-G3); from the latter it also removes oligosaccharides with higher molecular weight (G5-G6).

Developing a research agenda for promoting physical activity in Brazil through environmental and policy change

Objective. To identify the highest priorities for research on environmental and policy changes for promoting physical activity (PA) in Brazil; to uncover any gaps between researchers' and practitioners' priorities; and to consider which tools, methods, collaborative strategies, and actions could be useful to moving a research agenda forward.Methods. This was a mixed-methods study (qualitative and quantitative) conducted by Project GUIA (Guide for Useful Interventions for Activity in Brazil and Latin America) in February 2010-January 2011. A total of 240 individuals in the PA field (186 practitioners and 54 researchers) were asked to generate research ideas; 82 participants provided 266 original statements from which 52 topics emerged. Participants rated topics by "importance" and "feasibility;" a separate convenience sample of 21 individuals categorized them. Cluster analysis and multidimensional scaling were used to create concept maps and pattern matches.Results. Five distinct clusters emerged from the concept mapping, of which " effectiveness and innovation in PA interventions" was rated most important by both practitioners and researchers. Pattern matching showed a divergence between the groups, especially regarding feasibility, where there was no consensus.Conclusions. The study results provided the basis for a research agenda to advance the understanding of environmental and policy influences on PA promotion in Brazil and Latin America. These results should stimulate future research and, ultimately, contribute to the evidence-base of successful PA strategies in Latin America.

Protease activity in extracellular products secreted in vitro by trophozoites of Giardia duodenalis

There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in pathogenesis of giardiasis. This report describes a preliminary characterization of the proteolytic activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). The protease activity of E/S products in conditioned medium by trophozoites of each strain was analyzed using substrate (gelatin and collagen) impregnated SDS-PAGE and hemoglobin assay. The protease characterization was based on inhibition assays including synthetic inhibitors. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted degradation of the substrates and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibition assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases although the presence of serine proteases was also indicated, mainly in the hydrolysis of hemoglobin.

Heterochromatic banding pattern in two Brazilian populations of Aedes aegypti

We analysed samples of Aedes aegypti from Sao Jose do Rio Preto and Franca (Brazil) by C-banding and Ag-banding staining techniques. C-banding pattern of Ae. aegypti from Sao Jose do Rio Preto examined in metaphase cells differed from Franca. The chromosomes 2, 3 and X showed centromeric C-bands in both populations, but a slightly stained centromeric band in the Y chromosome was observed only in Sao Jose do Rio Preto. In addition, the X chromosome in both populations and the Y chromosome of all individuals from Sao Jose do Rio Preto showed an intercalary band on one of the arms that was absent in Franca. An intercalary, new band, lying on the secondary constriction of chromosome 3 was also present in mosquitoes of both populations. The comparison of the present data with data in the literature for Ae. aegypti from other regions of the world showed that they differ as to the banding pattern of sex chromosomes and the now described intercalary band in chromosome 3. The observations suggested that the heterochromatic regions of all chromosomes are associated to constitute a single C-banded body in interphase cells. Ag-banding technique stained the centromeric regions of all chromosomes (including the Y) and the intercalary C-band region of the X chromosome in both populations. As Ae. aegypti populations are widespread in a great part of the world, the banding pattern variations indicate environmental interactions and may reveal both the chromosome evolutionary patterns in this species and the variations that may interfere with its vector activity.

A novel biological activity for galectin-1: Inhibition of leukocyte-endothelial cell interactions in experimental inflammation

Galectin-1 (Gal-1), the prototype of a family of β -galactoside-binding proteins, has been shown to attenuate experimental acute and chronic inflammation. In view of the fact that endothelial cells (ECs), but not human polymorphonuclear leukocytes (PMNs), expressed Gal-1 we tested here the hypothesis that the protein could modulate leukocyte-EC interaction in inflammatory settings. In vitro, human recombinant (hr) Gal-1 inhibited PMN chemotaxis and trans-endothelial migration. These actions were specific as they were absent if Gal-1 was boiled or blocked by neutralizing antiserum. In vivo, hrGal-1 (optimum effect at 0.3 μg equivalent to 20 pmol) inhibited interleukin-1β-induced PMN recruitment into the mouse peritoneal cavity. Intravital microscopy analysis showed that leukocyte flux, but not their rolling velocity, was decreased by an anti-inflammatory dose of hrGal-1. Binding of biotinylated Gal-1 to resting and post-adherent human PMNs occurred at concentrations inhibitory in the chemotaxis and transmigration assays. In addition, the pattern of Gal-1 binding was differentially modulated by PMN or EC activation. In conclusion, these data suggest the existence of a previously unrecognized function of Gal-1, that is inhibition of leukocyte rolling and extravasation in experimental inflammation. It is possible that endogenous Gal-1 may be part of a novel anti-inflammatory loop in which the endothelium is the source of the protein and the migrating PMNs the target for its anti-inflammatory action.

Pattern of macrophage activation in yersinia-resistant and yersinia-susceptible strains of mice

Th1 cells, in cooperation with activated macrophages, are required to overcome Yersinia enterocolitica infection in mice. The pathway macrophages utilize to metabolize arginine can alter the outcome of inflammation in different ways. The objective of this study was to verify the pattern of macrophages activation in Y. enterocolitica infection of BALB/c (Yersinia-susceptible) and C57BL/6 (Yersinia-resistant) mice. Both strains of mice were infected with Y. enterocolitica O:8 WA 2707. Peritoneal macrophages and spleen cells were obtained on the 1st, 3rd and 5th day post-infection. The iNOS and the arginase activities were assayed in supernatants of macrophage cultures, by measuring their NO/citrulline and ornithine products, respectively. TGFβ-1 production was also assayed. The Th1 and Th2 responses were evaluated in supernatants of lymphocyte cultures, by IFN-γ and IL-4 production. Our results showed that in the early phase of Y. enterocolitica infection (1st and 3rd day), the macrophages from C57BL/6 mice produced higher levels of NO/citrulline and lower levels of ornithine than macrophages from BALB/c mice. The infection with Y. enterocolitica leads to an increase in the TGF-β1 and IL-4 production by BALB/c mice and to an increase in the IFN-γ levels produced by C57BL/6 mice. These results suggest that Y. enterocolitica infection leads to the modulation of M1 macrophages in C57Bl/6 mice, and M2 macrophages in BALB/c mice. The predominant macrophage population (M1 or M2) at the 1st and 3rd day of infection thus seems to be important in determining Y. enterocolitica susceptibility or resistance.

Leg muscles recruitment pattern in soccer players and active individuals during isometric contractions

This study aimed to compare the torque, torque ratio (Hamstrings:Quadriceps - H:Q), electromyographic (EMG) activity and EMG ratio (knee flexors:knee extensors EMG) in soccer players (SG, N=10) and active subjects (AG, N=10). Subjects performed three maximal voluntary isometric knee extensions and flexions at 45° and 90° to determine the peak torque and EMG activity. Torque and EMG activity of the knee flexor (biceps femoris [BF] and semitendinosus [ST]) were divided by the torque and EMG activity of the knee extensor (vastuls lateralis [VL] and rectus femoris [RF]) to calculate torque ratios (H:Q) and EMG ratios (BF:VL, BF:RF, ST:VL, ST:RF). The flexion torque was significantly higher for SG (p<0.05) in 45° and 90°. EMG activity for SG was significantly higher in agonist contractions for VL, RF and ST, and significantly lower in antagonist contractions for RF and ST when compared to AG Torque and EMG ratios were similar between groups and there were good correlations between torque ratio and BF:VL ratio (r=0.71, p=0.02) and BF:RF ratio (r=0.81, p=0.004) at 45. The EMG results could overestimate the joint balance calculated using torque ratios. Differences in recruitment pattern between soccer players and non-athletes can be related to the training routines and the EMG ratios presents applicable in trained populations.

MMP-2 and MMP-9 localization and activity in the female prostate during estrous cycle

The gerbil female prostate undergoes morphological and physiological changes resulting from hormonal fluctuations that occur during the reproductive cycle. These repetitive cycles of glandular growth and regression are followed by an extensive reconstruction and remodeling of prostate stroma throughout the reproductive life of the female gerbil. The objective of this study was to evaluate the effect that the hormonal fluctuations of the reproductive cycle have on the stromal remodeling and the expression and activity of matrix metalloproteinases MMP-2 and -9 in the adult female gerbil prostate. For this, serological, ultrastructural, immunohistochemical and biochemical methods were employed. The results showed that the major stromal alteration coincide with the peak of estradiol, which occurs in estrus, and with the peak of progesterone, occurring during diestrus II. MMP-2 and -9 presented a similar pattern of expression and activity during estrous cycle. The estrus was the phase of greater expression and activity of MMP-2 and -9. On the other hand, in DI and DII, the tissue expression and activity of MMP-2 and -9 was very weak. These results are important since they suggest the involvement of estradiol and progesterone in regulating the expression and activity of MMP-2 and -9 in adult gerbil female prostate. © 2011 Elsevier Inc.

Do Atopobium vaginae, Megasphaera sp. and Leptotrichia sp. change the local innate immune response and sialidase activity in bacterial vaginosis?

Objectives: To investigate if the participation of Atopobium vaginae, Megasphaera sp. and Leptotrichia sp. in the bacterial community of bacterial vaginosis (BV) is associated with distinct patterns of this condition. Methods: In this cross-sectional controlled study, 205 women with BV and 205 women with normal flora were included. Vaginal rinsing samples were obtained for measuring the levels of pro-inflammatory cytokines and bacterial sialidases. Real-time PCR was used to quantify the BV-associated bacteria and to estimate the total bacterial load using the 16S rRNA. Principal component analysis (PCA) using the measured parameters was performed to compare the BV samples with lower and higher loads of the species of interest. Results: Higher bacterial load (p<0.001), levels of interleukin 1-β (p<0.001) and sialidase activity (p<0.001) were associated with BV. Women with BV and higher relative loads of A vaginae, Megasphaera sp. and Leptotrichia sp. presented increased sialidase activity, but unchanged cytokine levels. PCA analysis did not indicate a different pattern of BV according to the loads of A vaginae, Megasphaera sp. and Leptotrichia sp. Conclusions: Greater participation of A vaginae, Megasphaera sp. and Leptotrichia sp. in vaginal bacterial community did not indicate a less severe form of BV; moreover, it was associated with increased sialidase activity.

Structure-activity relationship of compounds which are anti-schistosomiasis active

O método do orbital molecular AM1 foi empregado para calcular um conjunto de descritores moleculares para vinte neolignanas sintéticas com atividade anti-esquistossomose. O método de reconhecimento de padrão (análise de componentes principais ACP, análise de conglomerados AC e análise de discriminante) foi utilizado para obter a relação entre a estrutura molecular e a atividade biológica. O conjunto de moléculas foi classificado em dois grupos de acordo com seus graus de atividade biológica. Estes resultados permitem que, projete-se racionalmente novos compostos, potenciais candidatos à síntese e à avaliação biológica.

Increment of Antimycobaterial Activity on Lichexanthone Derivatives

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Transcriptional Activity, Chromosomal Distribution and Expression Effects of Transposable Elements in Coffea Genomes

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