Identification and characterization of a novel factor XIIa inhibitor in the hematophagous insect, Triatoma infestans (Hemiptera : Reduviidae)

Recently, we have cloned several Kazal-type serine protease inhibitors from the midgut of the Triatoma infestans bug. A single gene composed of multi Kazal-type domains, in tandem, encodes these inhibitors. in this work, we describe the purification and characterization of recombinant infestins 3-4 and 4, which are potent factor XIIa inhibitors (K-i = 67 pM and 128 pM, respectively). We also identified the first native factor XIIa inhibitor from a hematophagous insect. the factor XIIa inhibitory activity of infestin 4 demonstrates extremely efficient anticoagulant activity, prolonging activated partial thromboplastin time by approximately 3 times. Our results suggest that infestins perform a very important role in the T. infestans midgut during meal acquisition and digestion by controlling blood coagulation by means of inhibiting thrombin and factor XIIa. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.Universidade Federal de São Paulo, Dept Biochem, BR-04044020 São Paulo, BrazilInst Butantan, Physiol Lab, BR-05504900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biochem, BR-04044020 São Paulo, BrazilWeb of Scienc

Thrombin Inhibitors from Different Animals

Venous and arterial thromboembolic diseases are still the most frequent causes of death and disability in high-income countries. Clinical anticoagulants are inhibitors of enzymes involved in the coagulation pathway, such as thrombin and factor Xa. Thrombin is a key enzyme of blood coagulation system, activating the platelets, converting the fibrinogen to the fibrin net, and amplifying its self-generation by the activation of factors V, VIII, and XI. Thrombin has long been a target for the development of oral anticoagulants. Furthermore, selective inhibitors of thrombin represent a new class of antithrombotic agents. For these reasons, a number of specific thrombin inhibitors are under evaluation for possible use as antithrombotic drugs. This paper summarizes old and new interests of specific thrombin inhibitors described in different animals

Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity

Saliva of blood-sucking arthropods contains a complex mixture of anti-haemostatic, anti-inflammatory and immune-modulator compounds. Among anti-haemostatic factors, there are anticoagulants, vasodilators and platelet aggregation inhibitors. Previous analyses of the sialotranscriptome of Aedes aegypti showed the potential presence of a Kazal-type serine protease inhibitor in the female salivary glands, carcass and also in the whole male, which inhibitor we named AaTI (A. aegypti thrombin inhibitor). Recently, we expressed and characterized rAaTI as a trypsin inhibitor, and its anticoagulant activity [1]. in this work we characterized the thrombin inhibition mechanism of rAaTI. Recombinant AaTI was able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. in contrast, AaTI Delta (rAaTI truncated form) and C-terminal AaTI acidic tail prolong only thrombin time. in the competition assay, rAaTI, AaTI Delta or C-terminal AaTI acidic tail thrombin interactions seem to be affected by heparin but not by hirudin, suggesting that rAaTI binds to thrombin exosite 2. Finally, the thrombin inhibition assay of rAaTI showed an uncompetitive inhibition mechanism. in conclusion, rAaTI can probably inhibit thrombin by interacting with thrombin exosite 2, and the interaction is not mediated by the AaTI C-terminal region, since the truncated AaTI Delta form also prolongs thrombin time. (C) 2010 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilInst Butantan, Lab Fisiopatol, BR-05503900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilFAPESP: 05/03514-9FAPESP: 07/56614-6CNPq: 470070/2004-8CNPq: 575829/2008-7Web of Scienc

Differential transcript profile of inhibitors with potential anti-venom role in the liver of juvenile and adult Bothrops jararaca snake

Background. Snakes belonging to the Bothrops genus are vastly distributed in Central and South America and are responsible for most cases of reported snake bites in Latin America. The clinical manifestations of the envenomation caused by this genus are due to three major activities-proteolytic, hemorrhagic and coagulant-mediated by metalloproteinases, serine proteinases, phospholipases A(2) and other toxic compounds present in snake venom. Interestingly, it was observed that snakes are resistant to the toxic effects of its own and other snake's venoms. This natural immunity may occur due the absence of toxin target or the presence of molecules in the snake plasma able to neutralize such toxins. Methods. In order to identify anti-venom molecules, we construct a cDNA library from the liver of B. jararaca snakes. Moreover, we analyzed the expression profile of four molecules-the already known anti-hemorrhagic factor Bj46a, one gamma-phospholipase A(2) inhibitor, one inter-alpha inhibitor and one C1 plasma protease inhibitor-in the liver of juvenile and adult snakes by qPCR. Results. The results revealed a 30-fold increase of gamma-phospholipase A(2) inhibitor and a minor increase of the inter-alpha inhibitor (5-fold) and of the C1 inhibitor (3-fold) in adults. However, the Bj46a factor seems to be equally transcribed in adults and juveniles. Discussion. The results suggest the up-regulation of different inhibitors observed in the adult snakes might be a physiological adaptation to the recurrent contact with their own and even other snake's venoms throughout its lifespan. This is the first comparative analysis of ontogenetic variation of expression profiles of plasmatic proteins with potential anti-venom activities of the venomous snake B. jararaca. Furthermore, the present data contributes to the understanding of the natural resistance described in these snakes.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Tecnologico (CNPq)INCT - Entomologia MolecularInst Butantan, Lab Herpetol, Sao Paulo, BrazilUniv Sao Paulo, Interunidades Biotecnol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Bioquim, Sao Paulo, BrazilUniv Fed Rio de Janeiro, Dept Bioquim, Rio De Janeiro, BrazilInst Nacl Ciencia & Tecnol Entomol Mol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Bioquim, Escola Paulista Med, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Bioquim, Sao Paulo, BrazilFAPESP: 2010/10266-5FAPESP: 2012/03657-8FAPESP: 2013/05357-4FAPESP: 2014/11108-0CNPq: 308780/2013-2Web of Scienc

A DESTINAÇÃO FINAL DOS RESÍDUOS SÓLIDOS URBANOS: ALTERNATIVAS PARA A CIDADE DE SÃO PAULO ATRAVÉS DE CASOS DE SUCESSO

The article studies public sanitation services - in as much as solid domestic waste is concerned - within the municipality of São Paulo and employs as base year, that of 2010. Highlights include issues related to the current waste management model in the region and solutions are proposed to those deemed most critical, utilizing as benchmarks, adequate treatments conducted at other globalized cities. The bibliographical review seeks to elucidate concepts that are most aligned with the theme under study. Information concerning the management of solid domestic waste and reverse logistics in the private sector was gathered, whilst findings as to impairments observed in the municipality are listed. Local and international success cases concerning waste disposal were studied with views to encountering sources of potential improvements. It is understood that the adequate destination of residues, by means of recycling, composting or generation of fuels, calls for the population´s commitment in terms of developing separation and waste disposal habits, municipal administration involvement - in as much as supplying the required resources is concerned to ensure progress of alternatives - and finally, cannot do without reverse logistics actions undertaken by producing companies, in full compliance with existing federal rulings. In the quest to expand discussions concerning solid waste, the article attempts to include this matter - in a more effective manner - within strategic and investment decisions taken by both the public and private sectors.O artigo estuda o serviço de limpeza pública dos resíduos sólidos domiciliares do município de São Paulo, Brasil, tendo como base o ano de 2010. Destaca problemas associados ao atual modelo de gestão dos resíduos paulistanos, e propõe soluções para as questões mais críticas, tendo como referencial o tratamento adequado dado por outras cidades globalizadas. A revisão bibliográfica procura elucidar os conceitos mais relevantes ao estudo deste tema. São levantadas informações a respeito da gestão dos resíduos sólidos domiciliares e das iniciativas de logística reversa pelo setor privado, listando os entraves observados nesse município. Casos nacionais e internacionais de sucesso no que tange a destinação dos resíduos foram estudados, objetivando encontrar fontes de potenciais melhorias. Conclui-se que a destinação adequada dos resíduos, através da reciclagem, compostagem ou geração de energia, necessita de engajamento da população com hábitos de separação e disposição de resíduos, de envolvimento da administração municipal, provendo recursos necessários para a evolução destas alternativas, e por último, não pode prescindir de ações da logística reversa pelas empresas produtoras, respeitando a legislação federal. Ao procurar ampliar a discussão sobre os resíduos sólidos, o artigo busca inserir essa questão, de forma mais significativa, nas decisões de estratégia e investimento do poder público e privado

Purification and characterization of patagonfibrase, a metalloproteinase showing α-fibrinogenolytic and hemorrhagic activities, from Philodryas patagoniensis snake venom

Venoms of Colubridae snakes are a rich source of novel compounds, which may have applications in medicine and biochemistry. In the present study, we describe the purification and characterization of a metalloproteinase (patagonfibrase), the first protein to be isolated fromPhilodryas patagoniensis (Colubridae) snake venom. Patagonfibrase is a single-chain protein, showing a molecular mass of 53,224 Da and an acidic isoelectric point (5.8). It hydrolyzed selectively the Aá-chain of fibrinogen and when incubated with fibrinogen or plasma, the thrombin clotting time was prolonged. Prominent hemorrhage developed in mouse skin after intradermal injection of patagonfibrase. When administered into mouse gastrocnemius muscle, it induced local hemorrhage and necrosis, and systemic bleeding in lungs. Patagonfibrase showed proteolytic activity toward azocasein, which was enhanced by Ca 2+ and inhibited by Zn2+, cysteine, dithiothreitol and Na2EDTA. Patagonfibrase impaired platelet aggregation induced by collagen and ADP. Thus, patagonfibrase may play a key role in the pathogenesis of disturbances that occur in P. patagoniensis envenomation, and may be used as a biological tool to explore many facets of hemostasis.Fil: Peichoto, María Elisa. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Teibler, Pamela. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Mackessy, S.. Univeristy Of Northem Colorado. School Of Biological Sciences; Estados UnidosFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas y Naturales y Agrimensura; ArgentinaFil: Acosta, Ofelia Cristina. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Gonçalves, L. R. C.. Governo Do Estado de Sao Paulo. Secretaria Da Saude. Instituto Butantan. Laboratorio de Fisiopatologia; BrasilFil: Tanaka Azevedo, A.. Governo Do Estado de Sao Paulo. Secretaria Da Saude. Instituto Butantan. Laboratorio de Fisiopatologia; BrasilFil: Santoro, M.. Governo Do Estado de Sao Paulo. Secretaria Da Saude. Instituto Butantan. Laboratorio de Fisiopatologia; Brasi

Resistance training with slow speed of movement is better for hypertrophy and muscle strength gains than fast speed of movement.

Repetition speed is an important variable during resistance training. However, the effects of different speeds on the muscular strength and hypertrophy in isotonic resistance training are not clear. The study compared fast speed with slow speed of isotonic resistance training on muscular strength and hypertrophy in well-trained adults. Twelve healthy adults were randomly assigned into two groups: fast speed (FS) and low speed (SS). Muscle hypertrophy was measured by an ultrasound examination of the cross-sectional area of the brachial biceps muscle. Muscular strength was verified by 1 RM test. To check the possible differences in strength and hypertrophy between pre and post training and between groups there were compared by two-way ANOVA for repeated measurements and the effect size (ES) was calculated. Improvement in the cross-sectional area (P=0.019) and muscular strength (P=0.021) in the SS group between pre and post training was verified. The SS group had bigger effect sizes than FS group for hypertrophy and strength from pre to post training. SS training was more effective to improve hypertrophy and muscle strength in well-trained adults.Univ Fed Sao Paulo, Grp Studies & Res Exercise Physiol, Santos, SP, BrazilPraia Grande Coll, Grp Studies Sci Phys Educ, Praia Grande, SP, BrazilUniv Fed Sao Paulo, Dept Human Movement Sci, Santos, SP, BrazilUniv Fed Sao Paulo, Grp Studies & Res Exercise Physiol, Santos, SP, BrazilUniv Fed Sao Paulo, Dept Human Movement Sci, Santos, SP, BrazilWeb of Scienc

What’s in a mass

This short essay pretends to make the reader reflect on the concept of biological mass and on the added value that the determination of this molecular property of a protein brings to the interpretation of evolutionary and translational snake venomics research. Starting from the premise that the amino acid sequence is the most distinctive primary molecular characteristics of any protein, the thesis underlying the first part of this essay is that the isotopic distribution of a protein's molecular mass serves to unambiguously differentiate it from any other of an organism's proteome. In the second part of the essay, we discuss examples of collaborative projects among our laboratories, where mass profiling of snake venom PLA2 across conspecific populations played a key role revealing dispersal routes that determined the current phylogeographic pattern of the species.Ministerio de Ciencia, Innovación y Universidades/[BFU2017-89103-P]/MICIU/EspañaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP

Horseradish and soybean peroxidases: comparable tools for alternative niches?

Horseradish and soybean peroxidases (HRP and SBP, respectively) are useful biotechnological tools. HRP is often termed the classical plant heme peroxidase and although it has been studied for decades, our understanding has deepened since its cloning and subsequent expression, enabling numerous mutational and protein engineering studies. SBP, however, has been neglected until recently, despite offering a real alternative to HRP: SBP actually outperforms HRP in terms of stability and is now used in numerous biotechnological applications, including biosensors. Review of both is timely. This article summarizes and discusses the main insights into the structure and mechanism of HRP, with special emphasis on HRP mutagenesis, and outlines its use in a variety of applications. It also reviews the current knowledge and applications to date of SBP, particularly biosensors. The final paragraphs speculate on the future of plant heme-based peroxidases, with probable trends outlined and explored

FAMÍLIA E RECÉM-NASCIDO: DIRETRIZES PARA UMA NOVA PRÁTICA DE ENFERMAGEM

The purpose of this research is to identify the nursing team’s contributions to the construction of family bonding with the newborn for the development of a new care guideline. It is a descriptive qualitative study, based in the Problem-based Methodology with the participation of 20 nursing professionals who belonged to the Woman and Newborn Health-care Unit. Data collection, by means of Education Practice Workshops, was carried out between October and November 2012. Three thematic categories emerged from the analysis and interpretation of the qualitative data obtained in the workshops: a) Nursing care contributions to the construction of the family bonding with the neonate; b) Nursing care limitations for the construction of the family bonding with the neonate; c) Nursing care guidelines for the construction of the family bonding with the neonate. It was evidenced that the reflection held in the practice workshops enabled the team participation in the development of guidelines in order to build the family bonding with the neonate. However, nursing care needs to integrate with the other sectors to strengthen the network building of institutional humanized care.El objetivo de la investigación fue identificar las contribuciones del equipo de Enfermería en la construcción del vínculo familiar con el recién nacido para el desarrollo de una nueva directriz de cuidado. Es un estudio cualitativo descriptivo, cuya base fue la Metodología de la Problematización, en que participaron 20 profesionales del equipo de Enfermería de la Unidad de la Mujer y del Recién nacido. Los datos fueron obtenidos por medio de Prácticas Educativas realizadas en los meses de octubre y noviembre de 2012. Del análisis e interpretación del substrato cualitativo obtenido en las prácticas, resultaron tres categorías temáticas: a) Contribuciones del cuidado de enfermería para construcción del vínculo familiar con el recién nacido; b) Limitaciones del cuidado de Enfermería para la construcción del vínculo familiar con el recién nacido;  c) Directrices para el cuidado de enfermería en la construcción del vínculo familiar con el recién nacido. Se evidenció que la reflexión realizada en las prácticas ha posibilitado la participación colectiva en el desarrollo de directrices para el cuidado de enfermería en la construcción del vínculo familiar con el recién nacido. Sin embargo, el cuidado de enfermería necesita integrarse a los demás sectores para potencializar la construcción en red de un cuidado institucional humanizado.O objetivo desta pesquisa foi identificar as contribuições da equipe de enfermagem na construção do vínculo familiar com o recém-nascido para o desenvolvimento de uma nova diretriz de cuidado. Estudo qualitativo descrito, ancorado na Metodologia da Problematização, do qual participaram 20 profissionais da equipe de enfermagem pertencentes à Unidade da Mulher e do Recém-nascido. A coleta de dados ocorreu por meio de Oficinas de Práticas Educativas realizadas nos meses de outubro e novembro de 2012. Da análise e interpretação do substrato qualitativo obtido nas oficinas, emergiram três categorias temáticas: a) Contribuições do cuidado de enfermagem para construção do vínculo familiar com o recém-nascido; b) Limitações do cuidado de Enfermagem para a construção do vínculo familiar com o recém-nascido e c) Diretrizes para o cuidado de enfermagem na construção do vínculo familiar com o recém-nascido. Evidenciou-se que a reflexão realizada nas oficinas práticas, possibilitou a participação coletiva no desenvolvimento de diretrizes para o cuidado de enfermagem na construção do vínculo familiar com o recém-nascido. Entretanto, o cuidado de enfermagem necessita integrar-se aos demais setores para potencializar a construção em rede de um cuidado institucional humanizado