Bothrops moojeni myotoxin-II, a Lys49-phospholipase A(2) homologue: An example of function versatility of snake venom proteins

MjTX-II, a myotoxic phospholipase A(2) (PLA(2)) homologue from Bothrops moojeni venom, was functionally and structurally characterized. The MjTX-II characterization included: (i) functional characterization (antitumoral, antimicrobial and antiparasitic effects); (ii) effects of structural modifications by 4-bromophenacyl bromide (BPB), cyanogen bromide (CNBr), acetic anhydride and 2-nitrobenzenesulphonyl fluoride (NBSF); (iii) enzymatic characterization: inhibition by low molecular weight heparin and EDTA; and (iv) molecular characterization: cDNA sequence and molecular structure prediction. The results demonstrated that MjTX-II displayed antimicrobial activity by growth inhibition against Escherichia coli and Candida albicans, antitumoral activity against Erlich ascitic tumor (EAT), human breast adenocarcinoma (SK-BR-3) and human T leukemia cells (JURKAT) and antiparasitic effects against Schistosoma mansoni and Leishmania spp., which makes MjTX-II a promising molecular model for future therapeutic applications, as well as other multifunctional homologous Lys49-PLA(2)S or even derived peptides. This work provides useful insights into the structural determinants of the action of Lys49-PLA2 homologues and, together with additional strategies, supports the concept of the presence of others bioactive sites distinct from the catalytic site in snake venom myotoxic PLA(2)s. (c) 2005 Elsevier B.V. All rights reserved.

Cloning and Identification of a Complete cDNA Coding for a Bactericidal and Antitumoral Acidic Phospholipase A(2) from Bothrops jararacussu Venom

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

Presence of Staphylococcus spp. and Candida spp. in the human oral cavity

A presença de leveduras do gênero Candida e Staphylococcus na cavidade bucal humana é de extrema importância, pois podem atuar como microbiota suplementar e em determinadas situações causar doença bucal ou sistêmica. O objetivo do presente trabalho foi estudar a prevalência de Candida spp. e Staphylococcus spp. na cavidade bucal humana. Enxagüe bucal foi coletado de 68 indivíduos segundo a técnica proposta por Samaranayake e MacFarlane e a seguir semeados em ágar Sabouraud dextrose com cloranfenicol e ágar Baird-Parker. Após crescimento, os microrganismos foram isolados e identificados através de provas bioquímicas. Os dados foram analisados através de análise de variância (ANOVA). Leveduras do gênero Candida foram encontradas em 61,76% dos indivíduos examinados, sendo C. albicans a mais frequentemente isolada. Staphylococcus spp. foram isolados em 95,60% das cavidades bucais, sendo 41 cepas (63%) coagulase-negativas. Das cepas coagulase-positivas, nove eram S. aureus, 11 S. hyicus, e quatro S. schleiferi subespécie coagulans. Não foi observada correlação entre as contagens (UFC) de Candida spp. e Staphylococcus spp. encontradas nos enxagües bucais dos indivíduos examinados.

Testosterone stimulates growth and secretory activity of the female prostate in the adult gerbil (Meriones unguiculatus)

The prostate of the female gerbil (Meriones unguiculatus) is similar to the human female prostate (Skene gland) and, despite its reduced size, it is functional and shows secretory activity. However, virtually nothing is known about its physiological regulation. This study was thus undertaken to evaluate the behavior of the gerbil female prostate in a hyperandrogenic condition. Adult females received subcutaneous injections of testosterone cypionate (1 mg/kg body weight every 48 h) up to 21 days. Circulating levels of testosterone and estradiol were monitored, and the prostate and ovaries subjected to structural and immunocytochemical analyses. The treatment resulted in sustained high levels of circulating testosterone, and caused a transient increase in estradiol. There was an increase in epithelial cell proliferation accompanied by significant reorganization of the epithelium and an apparent reduction in secretory activity, followed by a progressive increase in luminal volume density and accumulation of secretory products. Immunocytochemistry identified the expression of androgen receptor and a prostate-specific antigen (PSA)-related antigen in prostatic epithelial cells. A circulating PSA-related antigen was also found, and its concentration showed strong negative correlation with circulating estrogen. Epithelial dysplasia was detected in the prostate of treated females. Analysis of the ovaries showed the occurrence of a polycystic condition and stromal cell hyperplasia. The results indicate that testosterone has a stimulatory effect on the female prostate, inducing epithelial cell proliferation, differentiation, secretory activity, and dysplasia. The results also suggest that prostatic growth and activity, polycystic ovaries, and ovarian stromal cell hyperplasia are related to a hyperandrogenic condition in females.

Penetration of 38% hydrogen peroxide into the pulp chamber in bovine and human teeth submitted to office bleach technique

This study evaluated the pulp chamber penetration of peroxide bleaching agent in human and bovine teeth after office bleach technique. All the teeth were sectioned 3 mm apical of the cement-enamel junction and were divided into 2 groups, A (70 third human molars) and B (70 bovine lateral incisors), that were subdivided into A1 and B1 restored by using composite resin, A2 and B2 by using glass ionomer cement, and A3 and B3 by using resin-modified glass ionomer cement; A4, A5, B4, and B5 were not restored. Acetate buffer was placed in the pulp chamber, and the bleaching agent was applied for 40 minutes as follows: A1-A4 and B1-B4, 38% hydrogen peroxide exposure and A5 and B5, immersion into distilled water. The buffer solution was transferred to a glass tube in which leuco crystal violet and horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined by spectrophotometer and converted into microgram equivalents of hydrogen peroxide. Data were submitted to analysis of variance and Dunnett, Kruskal-Wallis, and Tukey tests (5%). A higher level of hydrogen peroxide penetrated into the pulp chamber in resin-modified glass ionomer cements in bovine (0.79 +/- 0.61 mu g) and human (2.27 +/- 0.41 mu g) groups. The bleaching agent penetration into the pulp chamber was higher in human teeth for any experimental situation. The penetration of the hydrogen peroxide depends on restorative materials, and under the conditions of this study human teeth are more susceptible to penetration of bleaching agent into the pulp chamber than bovine teeth.

Desnaturação de oxihemoglobinas humanas A, A2 e S pelo metodo de tampão isopropanol

Human oxyhaemoglobin A and A2 from normal individuals and oxyhaemoglobin S from patients with sickle cell anaemia and sickle cell trait were studied using Isopropanol/buffer method at 37°C and 40°C. Hb S was less stable than Hb A, whereas Hb A2 was considerably more stable than either. Denaturation of Hb S was dependent on temperature and its concentration. Between the patients with sickle cell trait it was not possible to verify the influence of the concentration probably due to the small range used (from 38% to 44%).

Efeito α-tomatina na proliferação celular, apoptose e expressão de RNAm dos genes APC, Ciclina A2, Catenina, CASP9, BAK, BAX e BCL-XL em células HT29

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão em Escherichia coli de uma proteína recombinante do gene A2 de Leishmania chagasi e utilização desta proteína no diagnóstico da leishmaniose visceral canina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

An assessment of the economic impact of climate change on the Agriculture, Coastal and Human Settlements and Health Sectors in Guyana

This report analyses the agriculture, coastal and human settlements and health sectors in Guyana to assess the potential economic impacts of climate change. The fundamental aim of this report is to assist with the development of strategies to deal with the potential impact of climate change on Guyana. It also has the potential to provide essential input for identifying and preparing policies and strategies to help bring the Caribbean sub-region closer to solving problems associated with climate change and attaining national and regional sustainable development goals. Some of the key anticipated manifestations of climate change for the Caribbean include elevated air and sea-surface temperatures, sea-level rise, possible changes in extreme events and a reduction in freshwater resources. The economic impact of climate change on the three sectors was estimated for the A2 and B2 IPCC scenarios until 2050 (agriculture and health sectors) and 2100 (coastal and human settlements sector). An exploration of various adaptation strategies was also undertaken for each sector using standard evaluation techniques. The study of the impact of climate change on the agriculture sector focused on three leading sub-sectors namely: sugar-cane, rice-paddy and fisheries. In estimating costs, the sugar sub-sector is projected to experience losses under A2 between US$ 144 million (at 4% discount rate) and US$300 million (1% rate); comparative statistics for rice are US$795 million and US$1577 million, respectively; while for fisheries, the results show that losses range from US$15 million (4% rate) and US$34 million (1% rate). In general, under the B2 scenarios, there are gains for sugar up to 2030 under all three discount rates while for rice the performance is somewhat better with gains realized under all three discount rates up to 2040. For fisheries, gains are forecasted under all three rates up to 2050, following marginal losses to 2020. In terms of the benefit-cost analysis conducted on selected adaptation measures under the A2 scenario, there were net benefits for all three commodities under all three discount rates. For the sugar-cane sub-sector these are: drainage and irrigation upgrade, purchase of new machinery for planting and harvesting, developing and replanting climate tolerant sugar-cane. The rice-paddy sub-sector will benefit from adaptive strategies, which include maintenance of drainage and irrigation systems, research and development, as well as education and training. Adaptation in the fisheries sub-sector must include measures such as, mangrove development and restoration and public education. The analysis of the coastal and human settlements sector has shown that based upon exposed assets and population, SLR can be classified as having the potential to create catastrophic conditions in Guyana. The main contributing factor is the concentration of socioeconomic infrastructure along the coastline in vulnerable areas.

An assessment of the economic impact of climate change on the coastal and human settlements, tourism and transport sectors in Barbados

This report analyses the coastal and human settlements, tourism and transport sectors in Barbados to assess the potential economic impact of climate change on the sectors. The fundamental aim of this report is to assist with the development of strategies to deal with the potential impact of climate change on Barbados. Some of the key anticipated manifestations of climate change for the Caribbean include elevated air and sea-surface temperatures, sea-level rise, possible changes in extreme events and a reduction in freshwater resources. The economic impact of climate change on the three sectors was estimated for the A2 and B2 IPCC scenarios until 2050 (tourism and transport sectors) and 2100 (coastal and human settlements sector). An exploration of various adaptation strategies was also undertaken for each sector using standard evaluation techniques. The analysis has shown that based upon exposed assets and population, SLR can be classified as having the potential to create potential catastrophe in Barbados. The main contributing factor is the concentration of socioeconomic infrastructure along the coastline in vulnerable areas. The A2 and B2 projections have indicated that the number of catastrophes that can be classified as great is likely to be increased for the country. This is based upon the possible effects of the projected unscheduled impacts to the economy both in terms of loss of life and economic infrastructure. These results arise from the A2 and B2 projections, thereby indicating that growth in numbers and losses are largely due to socioeconomic changes over the projection period and hence the need for increased adaptation strategies. A key adaptation measure recommended is for the government of Barbados to begin reducing the infrastructure deficit by continuously investing in protective infrastructure to decrease the country’s vulnerability to changes in the climate. With regard to the tourism sector, it was found that by combining the impacts due to a reduction in tourist arrivals, coral reef loss and SLR, estimated total economic impact of climate change is US $7,648 million (A2 scenario) and US $5,127 million (B2 scenario). An economic analysis of the benefits and costs of several adaptation options was undertaken to determine the cost effectiveness of each one and it was found that four (4) out of nine (9) options had high cost-benefit ratios. It is therefore recommended that the strategies that were most attractive in terms of the cost-benefit ratios be pursued first and these were: (1) enhanced reef monitoring systems to provide early warning alerts of bleaching events; (2) artificial reefs or fish-aggregating devices; (3) development of national adaptation plans (levee, sea wall and boardwalk); (4) revision of policies related to financing carbon neutral tourism; and (5) increasing recommended design wind speeds for new tourism-related structures. The total cost of climate change on international transportation in Barbados aggregated the impacts of changes in temperature and precipitation, new climate policies and SLR. The impact for air transportation ranges from US$10,727 million (B2 scenario) to US$12,279 million (A2 scenario) and for maritime transportation impact estimates range from US$1,992 million (B2 scenario) to US$2,606 million (A2 scenario). For international transportation as a whole, the impact of climate change varies from US$12,719 million under the B2 scenario to US$14,885 million under the A2 scenario. Barbados has the institutions set up to implement adaptive strategies to strengthen the resilience of the existing international transportation system to climate change impacts. Air and sea terminals and facilities can be made more robust, raised, or even relocated as need be, and where critical to safety and mobility, expanded redundant systems may be considered.

An assessment of the economic impact of climate change on the coastal and human settlements Sector in Barbados

This study assesses the potential economic impact of climate change on coastal human settlements in the Caribbean, with specific reference to Barbados, and evaluates the costs and benefits of undertaking various adaptation strategies. The aim is to assist Caribbean territories in developing the strategies and capacity needed to deal with the potential impact of severe weather events that are anticipated to occur with increased frequency and intensity as a result of climate change. Some of the key anticipated manifestations of climate change for the Caribbean include elevated air and sea-surface temperatures, sea-level rise, possible changes in extreme events and a reduction in freshwater resources.This research focuses on how human settlements distributed along the coast of Guyana, especially those in low elevation coastal zones (LECZ)are affected by these impacts. Focusing on three potential transmission sources - sea-level rise, stronger storm hazards and increased precipitation – the study considers the vulnerability of populations in the LECZ areas and estimates the overall threat posed by climate change to coastal populations and infrastructure. Vulnerability to climate change (measured as exposed assets) was estimated for four emission scenarios as outlined by the Special Report on Emissions Scenarios (SRES), namely the A1, A2, B1 and B2 scenarios for the period 2010 to 2100 and as detailed by the Intergovernmental Panel on Climate Change (IPCC), using global circulation models (GCM) and storm surge hazard maps.

Metal ions bound to the human milk immunoglobulin A: Metalloproteomic approach

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

Pseudechis guttatus venom proteome: Insights into evolution and toxin clustering

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Carboxypeptidases A1 and A2 from the perfusate of rat mesenteric arterial bed differentially process angiotensin peptides

Here we report the isolation of carboxypeptidases A1 and A2 (CPA1 and CPA2) from the rat mesenteric arterial bed perfusate, which were found to be identical with their pancreatic counterparts. Angiotensin (Ang) I, Ang II, Ang-(1-9) and Ang-(1-12) were differentially processed by these enzymes, worthy mentioning the peculiar CPA1-catalyzed conversion of Ang II to Ang-(1-7) and the CPA2-mediated formation of Ang I from Ang-(1-12). We detected gene transcripts for CPA1 and CPA2 in mesentery and other extrapancreatic tissues, indicating that these CPAs might play a role in the renin-angiotensin system in addition to their functions as digestive enzymes. (C) 2011 Elsevier Inc. All rights reserved.

Anti-snake venom activities of extracts and fractions from callus cultures of Sapindus saponaria

Context: Sapindus saponaria L. (Sapindaceae) bark, root, and fruits are used as sedatives and to treat gastric ulcer and also demonstrate diuretic and expectorant effects. Objective: The anti-snake venom properties of callus of S. saponaria are investigated here for the first time. Materials and methods: In vitro cultivated callus of Sapindus saponaria were lyophilized, and the extracts were prepared with different solvents, before submitting to phytochemical studies and evaluation of the anti-ophidian activity. Crude extracts were fractionated by liquid-liquid partition and the fractions were monitored by thin layer chromatography (TLC). Subsequently, anti-ophidian activities were analyzed toward Bothrops jararacussu Lacerda (Viperidae), B. moojeni Hoge (Viperidae), B. alternates Dumeril (Viperidea) and Crotalus durissus terrificus Lineu (Viperidae) venoms and isolated myotoxins and phospholipase A(2) (PLA(2)). Results: Fractions A1, A2 and the extract in MeOH:H2O (9:1) significantly inhibited the toxic and pharmacological activities induced by snake venoms and toxins, when compared to other extracts and fractions. The lethal, clotting, phospholipase, edema-inducing, hemorrhagic and myotoxic activities were partially inhibited by the different extracts and fractions. TLC profiles of the crude extracts (B and C) and fractions (A1 and A2) showed beta-sitosterol and stigmasterol as their main compounds. Stigmasterol exhibited inhibitory effects on enzymatic and myotoxic activities of PLA(2). Discussion and conclusion: Sapindus saponaria extracts and fractions presented anti-ophidian activity and could be used as an adjuvant to serum therapy or for its supplementation, and in addition, as a rich source of potential inhibitors of enzymes involved in several pathophysiological human and animal diseases.