Vanadium distribution, lipid peroxidation and oxidative stress markers upon decavanadate in vivo administration

The contribution of decameric vanadate species to vanadate toxic effects in cardiac muscle was studied following an intravenous administration of a decavanadate solution (1 mM total vanadium) in Sparus aurata. Although decameric vanadate is unstable in the assay medium, it decomposes with a half-life time of 16 allowing studying its effects not only in vitro but also in vivo. After 1, 6 and 12 h upon decavanadate administration the increase of vanadium in blood plasma, red blood cells and in cardiac mitochondria and cytosol is not affected in comparison to the administration of a metavanadate solution containing labile oxovanadates. Cardiac tissue lipid peroxidation increases up to 20%, 1, 6 and 12 h after metavanadate administration, whilst for decavanadate no effects were observed except 1 h after treatment (+20%). Metavanadate administration clearly differs from decavanadate by enhancing, 12 h after exposure, mitochondrial superoxide dismutase (SOD) activity (+115%) and not affecting catalase (CAT) activity whereas decavanadate increases SOD activity by 20% and decreases ( 55%) mitochondrial CAT activity. At early times of exposure, 1 and 6 h, the only effect observed upon decavanadate administration was the increase by 20% of SOD activity. In conclusion, decavanadate has a different response pattern of lipid peroxidation and oxidative stress markers, in spite of the same vanadium distribution in cardiac cells observed after decavanadate and metavanadate administration. It is suggested that once formed decameric vanadate species has a different reactivity than vanadate, thus, pointing out that the differential contribution of vanadium oligomers should be taken into account to rationalize in vivo vanadate toxicity

RISCOS FÍSICOS E QUÍMICOS EM LABORATÓRIO DE ANÁLISES CLÍNICAS DE UMA UNIVERSIDADE

Study design : exploratory, case study. Objectives: to identify the physical and chemical risks faced by workers of a University Clinical Analysis Laboratory; study the occurrence, types, and causes of possible accidents and assess the workers’ degree of knowledge and use of protection measures. Methodology: data were collected from with lab workers using a selfadministered questionnaire, after receiving instructions about the characterization of physical and chemical risks according to the Ministry of Labor. Results: although workers consider they were exposed to the referred risks, no occupational accidents were detected and the workers used preventive measures for their team’s safety as well as their own. The findings also led to reflections about the observed reality, such as regular and continuing education, law applicability, decrees and regulations emitted by competent departments and the importance of adopting preventive measures. Conclusions: the revealed diagnosis makes it possible to create a local intervention plan and draws attention to the importance of realizing that workers are fallible human beings who face difficulties and pressures that can affect their professional performance.Modelo do estudo: exploratório, do tipo estudo de caso. Objetivos: identificar os riscos físicos e químicos a que estão expostos os trabalhadores do Laboratório de Análises Clínicas de uma Universidade; investigar a ocorrência, os tipos e as causas de possíveis acidentes e avaliar o grau de conhecimento e o uso de medidas de proteção pelos mesmos. Metodologia: os dados foram obtidos junto aos oito trabalhadores do laboratório, através de questionário respondido pelos próprios participantes, após orientações prévias acerca da caracterização dos riscos físicos e químicos preconizados pelo Ministério do Trabalho. Resultados: os resultados demonstraram que, apesar de os trabalhadores considerarem que estavam expostos aos referidos riscos, não foi detectado nenhum acidente de trabalho e que os mesmos fazem uso de medidas preventivas para sua segurança e a da equipe de trabalho. Possibilitaram também reflexões acerca da realidade encontrada, como formação e educação continuada, aplicabilidade de leis, decretos e portarias emitidas por órgãos competentes e da importância da adoção de medidas preventivas. Conclusões: o diagnóstico revelado viabiliza a elaboração de um plano de intervenção local e chama a atenção para a importância de se atentar que o trabalhador é um ser humano falível e defronta-se com dificuldades e pressões que podem interferir no seu desempenho profissional

Application of an Ionic Liquid in the Microwave Assisted Extraction of Cytotoxic Metabolites from Fruits of Schinus terebinthifolius Raddi (Anacardiaceae)

This work reports the application of an ionic liquid (1-butyl-3-methylimidazolium bromide, BMImBr) in the microwave assisted extraction (MAE) of metabolites from fruits of Schinus terebinthifolius. Dried fruits were individually extracted using BMImBr: H2O 1: 1, v/v (experiment 1) and pure H2O (experiment 2) by MAE (10 min at 60 degrees C). After partition using EtOAc, the yield to experiment 1 was about 23% while to experiment 2 was 0.1%. The EtOAc fraction obtained from experiment 1 was purified by chromatographic methods to afford 3-oxotirucalla-7,24Z-dien27- oic acid, 3a-hydroxytirucalla-7,24Z-dien-27-oic acid, 3 alpha-acetoxytirucalla-7,24Z-dien-27-oic acid, gallic acid, and ethyl gallate, being the first occurrence of the third compound as natural product. Cytotoxic activity was evaluated in vitro against cancer cell lines (A2058, HeLa, SiHa, HCT, SKBR-3, U87, and B16F2Nex2), being 3 alpha-acetoxytirucalla-7,24Z-dien-27-oic acid the more active metabolite with IC50 ranging from 10.9 +/- 1.3 to 17.3 +/- 1.4 mu g mL(-1), lower than that determined to positive control cisplatin.FAPESPCAPESCNPqInstituto de Ciências Ambientais, Químicas e Farmacêuticas, Universidade Federal de São Paulo (UNIFESP), 09972-270 Diadema-SP, BrazilDepartamento de Micro, Imuno e Parasitologia, Universidade Federal de São Paulo (UNIFESP), 04023-062 São Paulo-SP, BrazilInstituto de Ciências Ambientais, Químicas e Farmacêuticas, Universidade Federal de São Paulo (UNIFESP), 09972-270 Diadema-SP, BrazilDepartamento de Micro, Imuno e Parasitologia, Universidade Federal de São Paulo (UNIFESP), 04023-062 São Paulo-SP, BrazilFAPESP: BIOTA 2011/51739-6FAPESP: 2015/11936-2Web of Scienc

Evaluation of the effect of hypothermia by cold water immersion on blood neutrophils and lymphocytes of rats submitted to acute exercise

O estresse sistêmico induzido pelo exercício libera substâncias bioativas determinantes da mobilização neutrofílica. A crioterapia diminui a reação inflamatória e atenua a elevação da perfusão sanguínea induzida pelo exercício. O objetivo deste trabalho foi analisar a influência da hipotermia decorrente da crioimersão corporal (CIC) imediata ao esforço físico agudo nas concentrações neutrofílicas e linfocíticas no sangue. Os ratos do grupo controle (AI) foram mantidos em repouso enquanto os do grupo AII foram submetidos ao protocolo de CIC a 10ºC por 10 minutos. Enquanto os animais dos grupos BI, BII, BIII e BIV realizaram o esforço físico agudo (EFA) em água a 31ºC durante 100 minutos com sobrecarga corpórea de 5% do peso corporal, os dos grupos CI, CII, CIII e CIV foram submetidos ao EFA seguido imediatamente de CIC. Nos grupos B e C, os animais foram sacrificados nos períodos de 06 (I), 12 (II), 24 (III) e 48 (IV) horas posteriores ao EFA. Através da microscopia óptica realizou-se a contagem dos neutrófilos e linfócitos. Utilizou-se do Teste T Student para análise estatística considerando-se nível de significância p < 0,05. Observou-se uma significativa neutrofilia nos grupos AII, BI, BII, BIII, BIV, CI, CII e CIII em relação a AI, diferentemente do grupo CIV, que apresentou quantidade de neutrófilos igual ao grupo controle. Os valores de linfócitos nos grupos BII, BIII, BIV, CI e CII foram significativamente menores do que AI, e nos grupos AII, BI, CIII e CIV foram iguais a AI. A neutrofilia e a linfopenia posteriores ao intenso exercício agudo são mantidas por 48 horas ou mais, porém, mediante a aplicação da crioimersão corporal imediata ao exercício, são normalizadas em 24 horas.Systemic stress induced by exercise increases bioactive substances in plasma which leads to neutrophilic mobilization. Cryotherapy causes a decrease in the inflammatory reaction and attenuates high blood perfusion after exercise. The objective of this work was to analyze the influence of cold water immersion (CWI) after acute exercise on neutrophil and lymphocyte mobilization. A control group of rats (AI) was kept at rest and a second group (AII) was submitted to CWI at 10º C for 10 minutes. The animals of Groups BI, BII, BIII and BIV were submitted to acute exercise which consisted in swimming in water at 31º C for 100 minutes with a load equivalent to 5% of the body weight. Groups CI, CII, CIII and CIV were submitted to CWI immediately after acute exercise. The animals were sacrificed at 6 (I), 12 (II), 24 (III) and 48 (IV) hours after the exercise and neutrophil and lymphocyte cells were counted for all groups by optic microscopy. The Student t-test was used for statistical analysis with a significance level of p< 0.05. A significant increase in the number of neutrophils was observed in Groups AII, BI, BII, BIII, BIV, CI, CII and CIII compared to AI. The neutrophil count of the CIV Group was similar to the Control Group. There was a significant drop in the number of lymphocytes in Groups BII, BIII, BIV, CI and CII when compared to Group AI. The lymphocyte count of Groups AII, BI, CIII and CIV were similar to the Control Group. The changes in neutrophil and lymphocyte counts caused by acute exercise were reverted to normal at 24 hours by cold water immersion

Areas of natural occurrence of melipona scutellaris Latreille, 1811(Hymenoptera: Apidae) in the state of Bahia, Brazil.

The bee Melipona scutellaris is considered the reared meliponine species with the largest distribution in the North and Northeast regions of Brazil, with records from the state of Rio Grande do Norte down to the state of Bahia. Considering the importance of this species in the generation of income for family agriculture and in the preservation of areas with natural vegetation, this study aimed at providing knowledge on the distribution of natural colonies of M. scutellaris in the state of Bahia. Literature information, interviews with stinglessbee beekeepers, and expeditions were conducted to confirm the natural occurrence of the species. A total of 102 municipalities showed records for M. scutellaris, whose occurrence was observed in areas ranging from sea level up to 1,200-meter height. The occurrence of this species in the state of Bahia is considered to be restricted to municipalities on the coastal area and the Chapada Diamantina with its rainforests. Geographic coordinates, elevation, climate and vegetation data were obtained, which allowed a map to be prepared for the area of occurrence in order to support conservation and management policies for the species

The grapevine uncharacterized intrinsic protein 1 (VvXIP1) is regulated by drought stress and transports glycerol, hydrogen peroxide, heavy metals but not water

A MIP (Major Intrinsic Protein) subfamily called Uncharacterized Intrinsic Proteins (XIP) was recently described in several fungi and eudicot plants. In this work, we cloned a XIP from grapevine, VvXIP1, and agrobacterium-mediated transformation studies in Nicotiana benthamiana revealed that the encoded aquaporin shows a preferential localization at the endoplasmic reticulum membrane. Stopped-flow spectrometry in vesicles from the aqy-null yeast strain YSH1172 overexpressing VvXIP1 showed that VvXIP1 is unable to transport water but is permeable to glycerol. Functional studies with the ROS sensitive probe CM-H(2)DCFDA in intact transformed yeasts showed that VvXIP1 is also able to permeate hydrogen peroxide (H2O2). Drop test growth assays showed that besides glycerol and H2O2, VvXIP1 also transports boric acid, copper, arsenic and nickel. Furthermore, we found that VvXIP1 transcripts were abundant in grapevine leaves from field grown plants and strongly repressed after the imposition of severe water-deficit conditions in potted vines. The observed downregulation of VvXIP1 expression in cultured grape cells in response to ABA and salt, together with the increased sensitivity to osmotic stress displayed by the aqy-null yeast overexpressing VvXIP1, corroborates the role of VvXIP1 in osmotic regulation besides its involvement in H2O2 transport and metal homeostasis.This work was supported by European Union Funds (FEDER/COMPETE Operational Competitiveness Programme) and Portuguese national Funds (FCT-Portuguese Foundation for Science and Technology): KBBE-2012-6-3117 "Inovinne", FCOMP-01-0124-FEDER-022692 and PTDC/AGR-ALI/100636/2008. HN (SFRH/BD/74257/2010) and APM (SFRH/BD/65046/2009) were supported by PhD grants from FCT. The Interuniversity Attraction Poles Programme-Belgian Science Policy (IAP7/29) and the Belgian French community ARC11/16-036 project.info:eu-repo/semantics/publishedVersio

High precision astrometry mission for the detection and characterization of nearby habitable planetary systems with the Nearby Earth Astrometric Telescope (NEAT)

(abridged) A complete census of planetary systems around a volume-limited sample of solar-type stars (FGK dwarfs) in the Solar neighborhood with uniform sensitivity down to Earth-mass planets within their Habitable Zones out to several AUs would be a major milestone in extrasolar planets astrophysics. This fundamental goal can be achieved with a mission concept such as NEAT - the Nearby Earth Astrometric Telescope. NEAT is designed to carry out space-borne extremely-high-precision astrometric measurements sufficient to detect dynamical effects due to orbiting planets of mass even lower than Earth's around the nearest stars. Such a survey mission would provide the actual planetary masses and the full orbital geometry for all the components of the detected planetary systems down to the Earth-mass limit. The NEAT performance limits can be achieved by carrying out differential astrometry between the targets and a set of suitable reference stars in the field. The NEAT instrument design consists of an off-axis parabola single-mirror telescope, a detector with a large field of view made of small movable CCDs located around a fixed central CCD, and an interferometric calibration system originating from metrology fibers located at the primary mirror. The proposed mission architecture relies on the use of two satellites operating at L2 for 5 years, flying in formation and offering a capability of more than 20,000 reconfigurations (alternative option uses deployable boom). The NEAT primary science program will encompass an astrometric survey of our 200 closest F-, G- and K-type stellar neighbors, with an average of 50 visits. The remaining time might be allocated to improve the characterization of the architecture of selected planetary systems around nearby targets of specific interest (low-mass stars, young stars, etc.) discovered by Gaia, ground-based high-precision radial-velocity surveys.Comment: Accepted for publication in Experimental Astronomy. The full member list of the NEAT proposal and the news about the project are available at http://neat.obs.ujf-grenoble.fr. The final publication is available at http://www.springerlink.co

Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology.

Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. The acquisition of the sialyl residue allows the parasite to avoid lysis by serum factors and to interact with the host cell. A major drawback to studying the sialylation kinetics and turnover of the trypomastigote glycoconjugates is the difficulty to identify and follow the recently acquired sialyl residues. To tackle this issue, we followed an unnatural sugar approach as bioorthogonal chemical reporters, where the use of azidosialyl residues allowed identifying the acquired sugar. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. Main sialyl acceptors were identified as mucins by biochemical procedures and protein markers. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. Notably, labeling for α(1,3)Galactosyl residues only partially colocalize with sialylated mucins, indicating that two species of glycosylated mucins do exist, which are segregated at the parasite surface. Moreover, sialylated mucins were included in lipid-raft-domains, whereas TS molecules are not. The location of the surface-anchored TS resulted too far off as to be capable to sialylate mucins, a role played by the shed TS instead. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Therefore, shedding of TS occurs via microvesicles instead of as a fully soluble form

Essential oil composition and antioxidant and antimicrobial properties of Campomanesia pubescens O. Berg, Native of Brazilian Cerrado

In this study, the essential oil composition, total contents of phenolics and proanthocyanidins, antioxidant and antimicrobial activities from different plant parts (root, stem, leaf and fruit) of Campomanesia pubescens (DC) O. Berg from Brazilian Cerrado are reported. The root essential oil is distinguished from others by having only one representative of monoterpenes (alpha-terpenylacetate). The aerial parts of C. pubescens are rich in volatile terpenes, as expected, especially in fruits whose essential oil contained approximately 80 % of monoterpenes. The essential oils showed antimicrobial activity against oral pathogens. The root essential oil showed the strongest inhibition against Fusobacterium nucleatum (ATCC 25586). The leaf extract presented the highest concentration of phenolic and proanthocyanidins compounds. The lowest concentration necessary for inhibition of DPPH to 50 % ranged between 6.6 ± 1.6 and 56.6 ± 2.3 μg/mL. The leaf extract exhibited the highest inhibition, close to BHT.Colegio de Farmacéuticos de la Provincia de Buenos Aire