Atividades peptidásicas têm sido relacionadas à artrite reumatoide (AR). Os mecanismos de ação do tratamento usual, o metotrexato (MTX), e de terapias alternativos da AR, como a acupuntura com veneno de abelhas (BV), não são completamente compreendidos. Para contribuir com o conhecimento da AR, o presente estudo avaliou parâmetros consagrados para AR e atividades de aminopeptidases básica (APB), neutra (APN) e dipeptidil peptidase 4 (DPP4), e da endopeptidase prolil oligopeptidase (POP) em amostras de animais controle e submetidos à indução de artrite por colágeno (CIA) com ou sem tratamento sistêmico com BV e/ou MTX, e em animais resistentes, bem como efeitos diretos do BV ou do MTX sobre as atividades de aminopeptidases sob estudo. Em conclusão: atividades aminopeptidásicas estão envolvidas na CIA e podem ser potencialmente úteis na avaliação diagnóstica e prognóstica da AR; os tratamentos com BV e/ou MTX são capazes de amenizar ou normalizar essas alterações nas aminopeptidases; o BV não tem efeito analgésico, mas é capaz de diminuir o TNF-<font face=\"Symbol\">a; o BV pode conter algum inibidor da APN, enquanto o MTX parece agir como um inibidor não seletivo de APB, APN e DPP4. / Peptidase activities have been related to rheumatoid arthritis (RA). The mechanisms of action of the usual treatment, methotrexate (MTX), and alternative therapies for RA, such as bee venom (BV) acupuncture, are not completely known. To contribute to the understanding of RA, this study evaluated parameters of RA and activities of basic (APB), neutral (APN) and dipeptidyl peptidase 4 (DPP4) aminopeptidases, and oligopeptidase prolyl endopeptidase (POP) in samples from rats with collagen-induced arthritis (CIA) and in control rats with or without systemic treatment with BV and /or MTX and in resistant animals, as well as the direct effects of BV or MTX on the aminopeptidase activities under study. In conclusion: aminopeptidase activities are involved in CIA and may be potentially useful in the diagnostic and prognostic evaluation of RA; treatment with BV and / or MTX are able to reduce or normalize these aminopeptidase alterations; BV has no analgesic effect, but it is able to decrease TNF-<font face=\"Symbol\">a; BV may contain some inhibitor of APN, while the MTX seems to act as a nonselective inhibitor of APB, APN and DPP4.

O veneno do peixe peçonhento Thalassophryne nattereri induz a formação de uma resposta imune protetora de longa duração caracterizada por células B produtoras de anticorpos de memória (Bmem) e de longa vida (ASC antibody secreting cells). A partir de uma combinação de abordagens in vivo e in vitro investigamos a participação dos linfócitos T de memória e moléculas sinalizadoras CD4 e CD28 sobre a diferenciação e manutenção das ASC e analisamos a relação hierárquica entre os 2 tipos de células B de memória. Demonstramos que as ASC requerem mediante a sua localização (tecido inflamado, baço ou medula óssea) ou a expressão de B220 a integração de diferentes sinais cognatos (BCR, linfócitos TcM e TeM e a sinalização CD4 ou CD28) ou solúveis (IL-17A e IL-23) para a sobrevivência e a amplificação da produção de anticorpos de memória. / Thalassophryne nattereri fish venom induces the formation of a protective immune response characterized by memory B cells (Bmem) and the long-lived antibody-secreting cells (ASC). From a combined in vivo and in vitro approaches, we investigated the participation of memory T lymphocytes and CD4 or CD28 signaling molecules on the differentiation and maintenance of ASC survival and analyzed the hierarchical relationship between the two types of memory B cells. We demonstrated that ASC require dependent by localization (inflamed tissue, spleen or bone marrow) or B220 expression the integration of different cognate signals (BCR, TcM and TeM lymphocytes, CD4 or CD28 signaling) or soluble (IL-17A and IL-23) to survival and amplification of the memory antibodies production.

O contato com as cerdas da lagarta de Premolis semirufa (Pararama) desperta sintomas da inflamação aguda e nos indivíduos poliacidentados pode causar deformidades comuns às sinovites crônicas (pararamose). No presente estudo foi mostrado que o extrato das cerdas da lagarta apresenta intensa atividade proteolítica, sendo capaz de ativar o Sistema Complemento, promover hidrólise de C3, C4 e C5 e a geração de anafilatoxinas. Análises cromatográficas do extrato permitiram o isolamento de uma serinoprotease de 82 kDa capaz de promover tais atividades. Em modelo murino, foi verificado que o extrato é capaz de induzir altos títulos de anticorpos, pronunciada reação inflamatória, ativação de linfócitos T e APCs, bem como produção de citocinas pró-inflamatórias. Os dados obtidos demonstram a existência, no extrato das cerdas da pararama, de várias enzimas que podem atuar em conjunto na geração e desenvolvimento das manifestações clínicas da pararamose. / The contact with the Premolis semirufas caterpillar bristles (Pararama) causes symptoms of the acute inflammation and, in the individuals after multiple accidents, joint deformities common to chronic synovitis (pararamose) can occur. In the current study it was shown that caterpillar bristles extract has intense proteolytic activity, being able to activate the Complement System, induce cleavage of C3, C4 and C5, and the generation of anaphylatoxins. Chromatographic analysis of the extract allowed the isolation of a serine protease with Mr of 82 kDa able to promote such activities. In murine model, it was demonstrated that the extract is able to induce high antibody titers, a pronounced inflammatory reaction, activation of T lymphocytes and APCs, as well as the generation of proinflammatory cytokines. The data obtained demonstrate the existence, in the pararama bristles extract, of numerous enzymes that can act together in the generation and development of clinical manifestations of pararamose.

As serpentes do gênero Crotalus contribuem com 7,7% dos acidentes ofídicos anuais no Brasil e apresentam a letalidade mais elevada. O veneno crotálico apresenta como componente majoritário a crotoxina, formada pelas sub-unidades fosfolipase A2 (PLA2) e crotapotina, responsável pelos efeitos neurotóxicos e miotóxicos observados nas vítimas de acidente. Nós resolvemos investigar novas estratégias de imunização no sentido de direcionar a produção de anticorpos contra as frações de maior toxicidade do veneno. Utilizamos cavalos como animais soroprodutores, divididos em três grupos experimentais, Animais foram imunizados com veneno de C. d. terrificus, crotoxina, ou PLA2. Os plasmas obtidos foram analisados por testes padrões contra diferentes antígenos crotálicos e comparados com antivenenos produzidos pelo Instituto Butantan. Os resultados obtidos mostram o reconhecimento cruzado do veneno das diferentes Crotalus por todos os antivenenos utilizados, e uma maior especificidade dos plasmas produzidos contra crotoxina e PLA2 pelos componentes de maior toxicidade. / The serpents from Crotalus genus contribute with 7.7% of the annual ophidic accidents in Brazil, and have the highest lethality. The crotalic venom shows crotoxin, formed by the sub-units phospholipase A2 (PLA2) and crotapotin, as a major component and responsible for the neurotoxic and myotoxic effects shown by the snakebite victims. We decided to investigate new immunizations strategies focusing in orientating the immune response against the most toxic fractions of the venom. We used horses as serum producers, divided in three groups. Animals were immunized with either C. d. terrificus venom, crotoxin or PLA2. The plasmas obtained were analyzed by standart immunochemical tests against different crotalic antigens and compared with antivenoms produced by Instituto Butantan. The results show cross-recognition of all Crotalus venoms by the antivenoms, and a higher specificity from the plasmas produced against crotoxin and PLA2 for the most toxic components.

O veneno da subespécie de cascavel Crotalus durissus terrificus (C.d.terrificus) altera a coagulação, tem ação neurotóxica, miotóxica e efeito imunossupressivo. O veneno de C.d.terrificus e a sua fração majoritária, crotoxina (CTX), inibem a resposta celular e humoral, sendo esse efeito independente da indução de morte celular. O objetivo foi investigar os mecanismos envolvidos na imunossupressão exercida pela CTX. A CTX induz a produção de IL-10, TGF-<font face=\"Symbol\">&#946 e prostaglandina E2 (PGE2) pelas células dos camundongos que a receberam. Menor secreção de IFN-<font face=\"Symbol\">&#947 e IL-12 foi observada nas culturas de células de camundongos imunizados e que receberam CTX. A CTX aumenta a expressão da enzima indoleamina 2,3 dioxigenase e a geração de células T reguladoras em camundongos imunizados com OVA e que receberam CTX. A CTX foi capaz de inibir a expressão de CD40, CD80, CD86 e de MHC de classe II em células de camundongos imunizados e nas células dendríticas (DCs) purificadas destes animais. Em culturas de DCs, a CTX induz alta secreção de IL-10, TGF-<font face=\"Symbol\">&#946 e PGE2 e menor de IL-12. / The venom of the rattlesnake Crotalus durissus terrificus (C.d.terrificus) changes the coagulation system and presents neurotoxic, myotoxic and immunosuppressive effect. The venom and its main fraction, crotoxin (CTX) inhibited both the cellular and humoral response. This effect is not due to induction of cell death. The objective was to investigate the immunosuppressive mechanisms of CTX. CTX induces production of IL-10, TGF-<font face=\"Symbol\">&#946 and prostaglandin E2 (PGE2) by cells of mice that received it. Lower secretion of IFN-<font face=\"Symbol\">&#947 and IL-12 was observed in the cultured cells from mice immunized that received CTX. CTX promotes increased expression of the enzyme indoleamine 2,3 dioxygenase and generation of regulatory T cells in mice immunized with OVA and receiving the toxin. CTX was able to inhibit the expression of CD40, CD80, CD86 and MHC II in mice immunized and in dendritic cells (DCs) purified from these animals. In cultures of DCs, CTX increased secretion of IL-10, TGF-<font face=\"Symbol\">&#946 and PGE 2 and lower IL-12.

Crotalfina é um peptídeo sintetizado a partir da sequência de um composto analgésico purificado do veneno de serpentes Crotalus durissus terrificus. Apesar da atividade opióide, ensaios moleculares indicam que esse peptídeo não se liga diretamente aos receptores opióides, sugerindo que a liberação de opióides endógenos sejam os responsáveis pela atividade analgésica. Baseado em dados da literatura que demonstram a estreita relação entre o sistema canabinóide e o sistema opióide, o objetivo deste projeto foi avaliar o possível efeito da crotalfina sobre o sistema canabinóide. Os resultados indicam que receptores canabinóides CB2 estão envolvidos na antinocicepção acarretada pela crotalfina, na vigência de hiperalgesia induzida por PGE2. Ainda, este efeito é dependente de liberação de opióides endógenos, particularmente dinorfina A, agonista endógeno de receptores opióides do tipo kappa, sendo essa liberação dependente da ativação de receptores CB2. / Crotalphine is a peptide synthesized based on the sequence of the analgesic compound purified from the Crotalus durissus terrificus snake venom. Despite the opioid activity, molecular assays indicate that this peptide does not directly bind to opioid receptors, suggesting that the endogenous opioid release could be responsible for analgesic activity. Based on data from literature demonstrating the close relationship between the cannabinoid and the opioid systems, the goal of this project was to evaluate the possible effect of crotalphine on the cannabinoid system. Results demonstrated that CB2 cannabinoid receptors are involved in the antinociception induced by crotalphine during PGE2-induced hyperalgesia. In agreement, this effect is dependent of the release of endogenous opioids, particularly dynorphin A, the endogenous agonist of kappa opioid receptors. This release is dependent of the CB2 receptor activation.

Une soixantaine d'espèces de poisons démersaux fréquentent les eaux sub-littorales de la baie de Douarnenez, de la rade de Brest, de la baie d'Audierne et de la proche Iroise. Elles s'y répartissent surtout en fonction de la nature du fond. L'étude de l'évolution saisonnière des captures a, cependant, montré une influence des conditions hydro-climatiques, en particulier en baie de Douarnenez. L'utilisation de l'analyse des correspondances sur les fréquences d'apparition des espèces permet de différentier les peuplements hivernaux de ceux d'été. Cette alternance est également en rapport avec les périodes de reproduction de leurs espèces caractéristiques. Sur les nourriceries intertidales, l'étude des caractères physico-chimiques de l'eau montre l'extrême dépendance de la répartition des espèces, en particulier des juvéniles de Pleuronectiformes, vis-à-vis des arrivées d'eau douce continentale et de l'exposition de ces estrans par rapport au large. L'évolution mensuelle des quantités capturées montre le même schéma, quel que soit le lieu ; un accroissement des prises lors de l'élévation de la température de l'eau à partir de mai, suivi d'une diminution traduisant une fuite en profondeur des jeunes poissons en novembre. L'analyse des structures démographiques tendrait à montrer une double origine géographique des jeunes plies au Nord de la zone étudiée. Par contre, chez Solea lascaris, le bi modalisme du groupe 0 pourrait être attribué à la confusion de deux espèces dont les périodes de ponte seraient distinctes. Enfin, l'étude des contenus stomacaux des principaux prédateurs montre la grande variété des proies consommées ainsi qu'une variabilité des régimes, particulièrement en fonction de la taille des poissons. Ces observations, complétées par une analyse éthologique des régimes, permettent la mise en évidence des aspects primordiaux de la relation prédateur-proie. Plusieurs indices montrent que la compétition interspécifique entre les prédateurs est faible.

Dans le cadre de ce travail, nous nous sommes intéressés à un concept innovant d'empoisonnement homogène des neutrons par insertion de terres rare (erbium et gadolinium) dans les gaines en alliage de zirconium pour les réacteurs à eau pressurisée. L'étude des équilibres entre phases des alliages erbium-zirconium et gadolinium-zirconium est indispensable comme préalable à la mise en oeuvre industrielle de ce procédé prometteur d'empoisonnement. Ce travail a consisté à déterminer expérimentalement le diagramme de phases du système erbium-zirconium. Nous avons, par le biais de différentes caractérisations, obtenu des données diagrammatiques. Avec celles-ci, nous proposons un nouveau tracé du diagramme de phases. Celui-ci est radicalement différent de celui disponible dans la littérature. Nous avons modélisé le système par l'approche CALPHAD. Nous avons également déterminé les limites de solubilité des solutions solides terminales du système gadolinium-zirconium. Les données obtenues expérimentalement sont en accord avec le tracé expérimental de la littérature et avec le modèle thermodynamique disponible. Afin de prendre en compte l'oxydation des gaines en service, nous nous sommes également intéressés aux systèmes erbium-oxygène-zirconium et gadolinium-oxygène-zirconium. Le premier système a fait l'objet d'une étude expérimentale. Nous avons mis en place un procédé de synthèse par métallurgie des poudres, incluant la synthèse de celles-ci à partir de métaux massifs. La caractérisation des échantillons ternaires nous a permis de proposer deux coupes isothermes (800°C et 1100°C). Pour le système gadolinium-oxygène-zirconium, nous avons prédit les équilibres entre phases à différentes températures à l'aide de calculs effectués à partir d'une base de données que nous avons construite avec les modèles thermodynamiques de la littérature des systèmes oxygène-zirconium, gadolinium-zirconium et sesquioxyde de gadolinium-zircone. Enfin, nous avons travaillé avec des alliages erbium-zirconium fabriqués en milieu industriel. Nous nous sommes intéressés à leurs propriétés mécaniques en traction, en lien avec leur microstructure. Nous avons mis en évidence l'effet durcissant de l'erbium, notamment à 325?°C

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This study aimed at using different types of food wastes as major sources of protein to replace the fish meal used in fish feeds to produce quality fish. The major objectives were to (1) investigate the variations of metalloid/metals, polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCPs) in the fish ponds (pond mud and water), and food wastes used as fish feeds; (2) analyze bioaccumulation and biomagnification of pollutants in the food chains; and (3) evaluate the potential health risks of exposure (to these pollutants) via dietary intake of fish fed with food waste feeds. The traditional fish farming model was used to culture low trophic level fish: a filter feeder (bighead, Aristichthys nobilis), a herbivore (grass carp, Ctenopharyngodon idellus) and a bottom feeder (mud carp, Cirrhina molitorella), which are more environmental friendly as they can utilize more solar energy. Furthermore, low-trophic level fish are less susceptible to the accumulation of toxic chemicals. Two types of food wastes (mainly containing cereal (Food Waste A) and meat waste meal (Food Waste B)) were used as the major source of protein to replace the fish meal in fish feed to culture fish. The concentrations of metalloid (arsenic (As)), metals (mercury (Hg), cadmium (Cd), chromium (Cr), lead (Pb), zinc (Zn), copper (Cu) and nickel (Ni)) in water, suspended particulate matter and sediment of the 3 experimental fish ponds located in Sha Tau Kok Organic Farm were monitored (bi-monthly during the first half year and tri-monthly during the second half year) and the results were similar to or lower than those in the commercial fish ponds around the Pearl River Delta (PRD) region. Results of the health risk assessments indicated that human consumption of grass carp (a herbivore) which fed food waste feed pellets would be safer than other fish species (mud carp, bighead carp and largemouth bass). There were no or lower magnifications, and low concentrations of metalloid/metals contained in the ponds indicated that the practice of traditional pond management by draining pond water regularly can provide a better fish pond habitat for birds and other wildlife. Furthermore, the use of food waste instead of fish meal (mainly consisted of contaminated trash fish) further reduced Hg accumulation in the cultured fish. During October 2011 - December 2012, the concentrations of PAHs and OCPs in three experimental fish ponds were monitored (bi-monthly during the first half year and tri-monthly during the second half year). The results were similar to or lower than those obtained in commercial fish ponds around the PRD region. The mean concentrations of .PAHs and .OCPs in sediment and fish collected from the experimental fish ponds during the 2nd half year (May 2012 to December 2012) were significantly higher (p<0.05) than those in the 1st half year (October 2011 to April, 2012). .PAHs and .DDTs in the two species of fish (grass carp and bighead carp) were significantly increased (p<0.05) with time, and PAHs and DDTs in grass carp and bighead carp fed with commercial fish feed pellets (control group) were significantly higher (p<0.05) than the fish fed with food waste pellets (Food Waste A and Food Waste B). Fruit, vegetables, bone meal and meat products were the major sources of PAHs and OCPs contamination for producing Food Waste A and Food Waste B. No significant increases in PAHs and DDTs concentrations with trophic levels were observed in the experimental ponds, showing that PAHs were not biomagnifed in the omnivorous food chains (plankton, grass carp, bighead carp and mud carp). DDTs were lower magnifications than those predatory food chains (plankton, trash fish, and largemouth bass) in farmed ponds. There was a very low cancer risk for PAHs and DDTs exerted on humans via consumption of bighead carp, grass carp and mud carp (fed with food waste and commercial pellets). Furthermore, the use of food waste instead of fish meal (mainly consisted of contaminated trash fish) further reduced accumulation of PAHs and DDTs in the cultured fish.PAHs and DDTs exerted on humans via consumption of bighead carp, grass carp and mud carp (fed with food waste and commercial pellets). Furthermore, the use of food waste instead of fish meal (mainly consisted of contaminated trash fish) further reduced accumulation of PAHs and DDTs in the cultured fish. The present results revealed that recycling of food waste for cultivating low trophic level fish (mainly bighead carp and grass carp) is feasible, which will also ease the disposal pressure of the large volume of food waste, a common problem encountered in densely populated cites such as Hong Kong.