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Agentes estressores em piscicultura intensiva afetam a condição fisiológica e o sistema imune dos peixes aumentando a susceptibilidade a infecções e causando prejuízos econômicos. Para contornar este problema, o uso de nutrientes moduladores do sistema imunológico na dieta, como o tocoferol, se tornam estratégia profilática para as pisciculturas assegurarem uma maior sobrevivência e consequente produtividade. O presente estudo visou avaliar a exigência nutricional do dourado, Salminus brasiliensis, por meio de um ensaio dose-resposta e aferir seu efeito imunomodulador. O delineamento utilizado foi o inteiramente casualizado, com sete tratamentos/dietas semi purificadas elaboradas com doses crescentes de vitamina E (0; 50; 100; 150; 200, 250 e 3500 mg kg-1α-tocoferol acetato) (n=4). Quinze juvenis de dourado foram alojados em caixas de polipropileno, em sistema de recirculação fechado com filtragem mecânica e biológica, trocador de calor com termostato e aeração forçada através de sopradores e pedras difusoras. Após serem alimentados por 30 dias com a dieta basal para depleção de vitamina E nas reservas corporais de vitamina, os animais foram pesados (20,04 ± 0,09 g), redistribuídos em 28 tanques (500 L; 15 peixes por tanque) e alimentados durante 78 dias até saciedade aparente (duas refeições diárias: 08h00m e 17h00m). Os peixes apresentaram diferença no ganho de peso, porém não de maneira dependente do tratamento de vitamina E. Após o período de alimentação os peixes foram desafiados com a bactéria Aeromonas hydrophila. Os parâmetros imunológicos foram aferidos antes e após o desafio e não houveram diferenças significativas para: atividade respiratória dos leucócitos, atividade da lisozima sérica, proteína e albumina total no soro e sobrevivência. Houve aumento da concentração das globulinas totais no soro após infecção em função do nível de vitamina E na dieta, e a dose ótima foi definida por meio da regressão segmentada: 151,07 mg kg-1 de vitamina E na ração. A exigência de vitamina E para o dourado foi determinada em 58,90 mg kg-1 pelas substâncias reativas ao ácido tiobarbiturico da fração mitocondrial do fígado por análise de regressão segmentada. / Stressors in intensive fish farming affect fish physiological conditions and immune system, promoting susceptibility to infections and causing economic losses. To overcome this issue, using immunomodulatory nutrients such as tocopherol on fish feeds may become a prophylactic strategy to fish farms to ensure a higher survival and productivity. The present study aimed at determining vitamin E requirement of dourado, Salminus brasiliensis, through a dose dependent trial and how the immune system responded to vitamin E crescent doses. The experiment design was randomly distributed in seven treatments (n=4) of semi purified diets (0; 50; 100; 150; 200; 250 and 3,500 mg kg-1). After exposing the fish during 30 days for tocopherol depletion, fifteen dourado juveniles (20.04 ± 0.09 g) were stocked in 500 L tanks in a closed loop system and fed twice a day until apparent satiation. Differences in weight gain were observed however not in a vitamin E dose dependent way. After feeding for 78 days fish were challenged with intraperitoneal bacterial infection (Aeromonas hydrophila). Immunological parameters were measured before and after infection and no significant differences were observed in: blood leukocyte burst respiratory activity, serum lysozyme activity, serum protein and albumin, and survival. However a higher total serum globulin was detected after infection and the best response determined through a broken-line regression was 151.07 mg kg-1 of vitamin E on fish feed. Dietary requirement of vitamin E was determined by broken-line regression from thiobarbituric reactive substances of liver mithocondrial fraction to be 58.90 mg/kg of α-tocopherol acetate.

1. INTRODUCTION The incidence of diseases whose pathogenesis chain includes changes in the functioning of the immune system is currently on the increase. The increasing prevalence of primary and secondary immunodeficiencies is a relevant issue in immunopathology. Secondary immunodeficiencies, caused by such factors as bacteria, viruses, helminths, chemical substances, medications, radiation, stress, etc., result in higher morbidity. Immunosuppression occurs in cases of oncological or allergic diseases, bronchial asthma, etc. immunosuppression is relevant in cases of malignant tumors whose incidence in Lithuania has been increasing with every year during the recent period. Oncological treatment involves the application of radiotherapy and chemotherapy with cytostatics that are strong immunosuppressants, and thus the restoration of the immune system with immunostimulants is becoming a relevant issue. Insufficient nutrition, disturbed nutrient resorption, negative nitrogen balance, chronic alcoholism, and drug abuse – all these factors result in the deceleration of cell proliferation and decreased T-lymphocyte counts and function. Immunodeficiency increases morbidity with infectious diseases, since the organism becomes incapable of the rapid destruction of the causative agents. Such conditions increase the probability of the spreading of the infection, and condition the increased activity of the opportunistic infections caused by adenoviruses, cytomegaloviruses, etc... [to full text]

Agentes estressores em piscicultura intensiva afetam a condição fisiológica e o sistema imune dos peixes aumentando a susceptibilidade a infecções e causando prejuízos econômicos. Para contornar este problema, o uso de nutrientes moduladores do sistema imunológico na dieta, como o tocoferol, se tornam estratégia profilática para as pisciculturas assegurarem uma maior sobrevivência e consequente produtividade. O presente estudo visou avaliar a exigência nutricional do dourado, Salminus brasiliensis, por meio de um ensaio dose-resposta e aferir seu efeito imunomodulador. O delineamento utilizado foi o inteiramente casualizado, com sete tratamentos/dietas semi purificadas elaboradas com doses crescentes de vitamina E (0; 50; 100; 150; 200, 250 e 3500 mg kg-1α-tocoferol acetato) (n=4). Quinze juvenis de dourado foram alojados em caixas de polipropileno, em sistema de recirculação fechado com filtragem mecânica e biológica, trocador de calor com termostato e aeração forçada através de sopradores e pedras difusoras. Após serem alimentados por 30 dias com a dieta basal para depleção de vitamina E nas reservas corporais de vitamina, os animais foram pesados (20,04 ± 0,09 g), redistribuídos em 28 tanques (500 L; 15 peixes por tanque) e alimentados durante 78 dias até saciedade aparente (duas refeições diárias: 08h00m e 17h00m). Os peixes apresentaram diferença no ganho de peso, porém não de maneira dependente do tratamento de vitamina E. Após o período de alimentação os peixes foram desafiados com a bactéria Aeromonas hydrophila. Os parâmetros imunológicos foram aferidos antes e após o desafio e não houveram diferenças significativas para: atividade respiratória dos leucócitos, atividade da lisozima sérica, proteína e albumina total no soro e sobrevivência. Houve aumento da concentração das globulinas totais no soro após infecção em função do nível de vitamina E na dieta, e a dose ótima foi definida por meio da regressão segmentada: 151,07 mg kg-1 de vitamina E na ração. A exigência de vitamina E para o dourado foi determinada em 58,90 mg kg-1 pelas substâncias reativas ao ácido tiobarbiturico da fração mitocondrial do fígado por análise de regressão segmentada. / Stressors in intensive fish farming affect fish physiological conditions and immune system, promoting susceptibility to infections and causing economic losses. To overcome this issue, using immunomodulatory nutrients such as tocopherol on fish feeds may become a prophylactic strategy to fish farms to ensure a higher survival and productivity. The present study aimed at determining vitamin E requirement of dourado, Salminus brasiliensis, through a dose dependent trial and how the immune system responded to vitamin E crescent doses. The experiment design was randomly distributed in seven treatments (n=4) of semi purified diets (0; 50; 100; 150; 200; 250 and 3,500 mg kg-1). After exposing the fish during 30 days for tocopherol depletion, fifteen dourado juveniles (20.04 ± 0.09 g) were stocked in 500 L tanks in a closed loop system and fed twice a day until apparent satiation. Differences in weight gain were observed however not in a vitamin E dose dependent way. After feeding for 78 days fish were challenged with intraperitoneal bacterial infection (Aeromonas hydrophila). Immunological parameters were measured before and after infection and no significant differences were observed in: blood leukocyte burst respiratory activity, serum lysozyme activity, serum protein and albumin, and survival. However a higher total serum globulin was detected after infection and the best response determined through a broken-line regression was 151.07 mg kg-1 of vitamin E on fish feed. Dietary requirement of vitamin E was determined by broken-line regression from thiobarbituric reactive substances of liver mithocondrial fraction to be 58.90 mg/kg of α-tocopherol acetate.

Le cancer est l’une des principales causes de mort dans le pays développés ; bien que les opérations chirurgicales, la radiothérapie et la chimiothérapie représentent aujourd’hui les principales options de traitement des patients souffrants de tumeurs malignes, leurs effets secondaires sévères ont ouvert la voie au développement de l’immunothérapie antitumorale. A part l’immunothérapie passive, qui est basée sur les anticorps ou tout autre composant du système immunitaire synthétisés en dehors du corps dont la potentielle menace de réactions immunes a été prouvée, nous avons concentré nos efforts sur l’immunothérapie active, qui réside dans la stimulation du système immunitaire du patient pour éradiquer sélectivement les cellules malignes. L’identification d’antigènes carbohydrates associés aux tumeurs (TACAs), surexprimés à la surface des cellules cancéreuses, a permis le développement de vaccins spécifiques à cet antigène. Il est connu depuis plus de 40 ans que la majorité des cancers chez l’homme sont caractérisés par une glycosylation aberrante. Les cellules tumorales peuvent surexprimer des versions tronquées d’oligosaccharides, une séquence terminale inhabituelle ou une augmentation de la sialylation des glycolipides et des glycoprotéines de surface. Un oligosaccharide d’une glycoprotéine tronqué peut rendre une partie de la chaîne principale du peptide, d’habitude caché par le sucre, plus accessible au système immunitaire. Parmi les différents TACAs, nous avons concentré notre attention sur les antigènes Tn et Tf, qui peuvent être trouvés sur des glycoprotéines comme MUC-1, surexprimés sur plus de 90% des carcinomes du sein. Bien que la conception de ces immunomodulateurs repose toujours sur des règles empiriques, il est important de déclencher à la fois la réponse humorale et cellulaire, ainsi qu’un effet de mémoire. Ce défi peut être relevé en combinant, sur une seule molécule, l’antigène carbohydrate exprimés à la surface des tumeurs (épitope des cellules B), les peptides capables de stimuler les cellules CD4+ et CD8+ (épitopes des cellules T) et un adjuvant, pour recueillir tous les éléments du système immunitaire au niveau du site d’injection et renforcer l’absorption des antigènes. De précédentes études faites dans notre groupe de recherche ont publié pour la première fois la synthèse et l’évaluation immunologique d’un prototype de vaccin anticancéreux à quatre composant capable d’induire une réponse immunitaire de longue durée sur des modèles murins. Dans mon travail de thèse, nous avons voulu synthétiser des prototypes de vaccin anticancéreux basés sur les TACAs avec des propriétés immunologiques accrues. Notre stratégie de conception a été guidée par (i) l’importance d’une haute densité de carbohydrates pour promouvoir une capture d’antigène plus efficace et un traitement par les cellules présentatrices d’antigène, et (ii) l’expression hétérogène des TACAs au cours de la maladie et parmi différents patients. En respectant ces deux aspects, il sera possible de déclencher une réponse immunitaire plus forte et à plusieurs facettes. / Cancer is one on the leading causes of death in developed countries; although surgical resection, direct irradiation and cytotoxic chemotherapy represent nowadays the main treatment options for patients suffering with malignancies, their severe side effects paved the way for the rise in popularity of antitumoral immunotherapy. Apart from passive immunotherapy, which is comprised of antibodies or other immune system components that are made outside of the body and has been shown to be associated to potentially life threatening immune reactions, we focused our efforts towards active immunotherapy, which purpose is stimulate the patient immune system to selectively eradicate malignant cells. The identification of tumor-associated carbohydrate antigens (TACAs) on the surface of cancer cells has allowed the development of antigen-specific vaccines. It has been known for over four decades that the majority of human cancers are characterized by aberrant glycosylation. Tumor cells may over-express truncated versions of oligosaccharides, unusual terminal oligosaccharide sequences, or increase sialylation of cell-surface glycolipids and O- and N-linked glycoproteins. A truncated oligosaccharide of a glycoprotein may render a part of the peptide backbone, which is normally shielded by the glycan, more accessible to the immune system. Among the assortment of TACAs we focussed our attention on Tn and TF-antigens, which can be found in membrane-bound glycoproteins like MUC-1, over-expressed in more than 90% of breast carcinomas. Although the design of such immuno-modulators still relies on empiric rules, it is noteworthy important to trigger both humoral and cellular responses, and a memory effect. This challenge can be achieved by combining, within a single molecule, carbohydrate antigen expressed on the surface of tumors (B-cell epitope), peptides capable to stimulate both CD4+ and CD8+ T-cells (T-cell epitopes) and an adjuvant, to gather immune system elements in the injection site and boost the antigen uptake. Previous studies of our research group reported for the first time the synthesis and immunological evaluation of a four-component anticancer vaccine prototype capable of inducing a long-lasting immune response in mice models. In my PhD work we aimed to synthesize TACA-based anticancer vaccine prototypes with improved immunological properties. The principles which guided our design strategies rely on (i) the importance of a high density of carbohydrate epitopes to promote a more effective antigen capture and processing by antigen-presenting cells, and (ii) the evidence of heterogenic expression patterns of TACAs during the course of the disease and among different individuals. Addressing these two aspects would provide a stronger and multifaceted immune response.

Host defense against infection and disease relies on the reciprocal communication between the immune and neuroendocrine systems where sex hormones exert negative and positive feedback actions on immune functions. Indeed, sex hormones have been implicated in gender dimorphic immune response and in the potentiation of immune-related disorders. The female hormone estrogen plays a role as an immunomodulator and may exert immunosuppressive and immunostimulatory effects. Though many studies focus on estrogen’s role in immunity within the female reproductive tract and autoimmunity, the modulatory effects of estrogen on vaccine responses are largely unexplored. The insufficient efficacy of some vaccines in certain target populations, as for example the elderly population, is well recognized. Hormones fluctuate throughout an individual’s life, and females in particular undergo several necessary reproductive (pregnancy and menopause) and lifestyle (oral contraceptive use) changes which involve sex hormones. Vaccine efficacy might be influenced by endogenous estrogen levels or by exogenous estrogen administration. Therefore, in the pursuit of improved vaccine efficacy, it is necessary to consider such hormonal factors and their contribution to immune status. We have studied estrogen’s role in modulation of vaccine responses using a mouse ovariectomy model where exogenous estrogen delivery can be controlled. Our studies included two different types of vaccines, a bacterial toxoid formulation and a bacterial secreted protein formulation. Results from these studies indicate that estrogen enhances vaccine-specific antibody production by likely supporting a general TH2 pathway and also modulates expression of genes encoding molecules critical in innate immune signaling and required for development of proper adaptive immune responses and antigen clearance through antibody-mediated mechanisms. The level at which estrogen modulates antibody responses appears to be dependent on the route of vaccine administration. The enhancement of specific humoral responses may involve mechanisms involving TLR2 and antibody Fc receptor expression on macrophages, cells that link innate and adaptive immune responses. Advances in our understanding of the relationship between sex hormones and the immune system may provide new insights into the mechanisms by which hormones act and thus may be exploited to guide the design of future vaccine strategies.

A Criptococose é uma enfermidade micótica sistêmica, subaguda ou crônica, que acomete a cavidade nasal, tecidos paranasais e pulmões do homem, animais domésticos e silvestres, podendo disseminar-se para o sistema nervoso central, olhos, pele e outros órgãos. Considerando as dificuldades terapêuticas no tratamento da micose em pequenos animais, incluindo toxicidade, desenvolvimento de resistência aos antifúngicos tradicionalmente utilizados e longos períodos de tratamento da enfermidade, o estudo objetivou avaliar a eficácia do imunomodulador ß (1-3) glucana isoladamente e em associação ao fluconazol no tratamento da criptococose experimental. Foram utilizados 100 camundongos (Mus musculus), cepa UFPel, albinos, os quais foram divididos em cinco grupos de 20 animais. O tratamento dos animais com criptococose experimental foi iniciado sete dias após a inoculação. O grupo Controle (G1) foi tratado com 0,1 ml de água destilada estéril, o grupo Fluconazol (G2) com 5 mg/kg de fluconazol, o grupo Fluconazol associado a ß (1-3) glucana (G3) com 5 mg/kg de fluconazol associado a 0,5 mg de ß (1-3) glucana, o grupo Glucana dose I (G4) com 0,5 mg de ß (1-3) glucana e o grupo Glucana dose II (G5) recebeu 0,25 mg de ß (1-3) glucana. Após acompanhamento clínico durante as seis semanas de tratamento os animais foram eutanasiados e necropsiados para avaliação anatomopatológica dos órgãos, quantificação das unidades formadoras de colônias fúngicas (UFCs), retroisolamento e avaliação histopatológica. Nas avaliações clínicas o G3 foi sempre superior ao G1 e G2, sendo que na última avaliação clínica individual, os animais pertencentes ao G3 não apresentavam nenhum sintoma clínico. O G2 teve um menor número de órgãos afetados e de alterações macroscópicas, seguido pelo G3, sendo que no primeiro não foram observadas lesões em órgãos-alvo, como pulmão e cérebro. O grupo com maior número de lesões e órgãos afetados foi o G1. Nos parâmetros retroisolamento e UFCs, o G3 foi superior aos outros tratamentos, seguido pelo G2, sendo que o pior grupo, ou seja, aquele que teve um maior número de isolamentos e maior quantificação de UFCs foi o G1. Conforme os resultados obtidos os tratamentos G2 e G3 foram os mais eficazes para a remissão da criptococose experimental, no entanto, G3 apresentou uma superioridade na maioria dos parâmetros avaliados. Portanto, de acordo com os resultados obtidos, a associação de fluconazol ao imunomodulador ß (1-3) glucana pode ser uma alternativa benéfica em considerável quantidade de pacientes com criptococose. / Cryptococcosis is a systematic sub acute or chronic mycotic condition that affects the nasal cavity, paranasal tissues and the lungs of humans, as well as domestic and wild animals, which can spread to the central nervous system, skin, and other organs. Considering the therapeutic difficulties in treating this mycosis in small animals, which include toxicity, resistance towards traditionally used antifungals, and the extended treatment of this condition, this study aimed to evaluate the ß (1-3) glucan immunomodulator efficacy when used both alone and in association with fluconazole in the treatment of experimental cryptococcosis. One Hundred albino UFPel strain mice (Mus musculus), divided into five groups of 20 animals each, were used. The treatment of these animals with experimental cryptococcosis was started seven days following inoculation. The control group (G1) was treated with 0,1 ml sterile distilled water. The fluconazole group (G2) was treated with 5 mg/kg fluconazole; the fluconazole associated with ß glucan (1-3) group (G3) was treated with 5 mg/kg fluconazole associated with 0,5 mg ß (1-3) glucan ; the group glucan dose I (G4) was treated with 0,5 mg ß (1-3) glucan, and the group glucan dose II (G5) was administered 0,25 mg ß (1-3) glucan. After the six-week treatment clinical follow-up, the aimals were euthanized and necropsied for anatomopathological evaluation of organs, quantification of fungal colony-forming units (FCUs), retro isolation and histopathological evaluation. In clinical evaluations, G3 was always superior to G1 and G2, and in the last individual clinical evaluation, G3 animals did not show any clinical symptoms. G2 had a smaller number of affected organs and microscopic alterations, followed by G3. In G2, target organ lesions, such as those in the lungs and brain, were not found. Considering retro isolation and FCU parameters, G3 was superior to other treatments, followed by G2; the worst group, that is, the one with the highest isolation occurrence and greatest FCU quantification, was G1. According to the results obtained, treatments G2 and G3 were the most efficient in experimental cryptococcosis remission; however, G3 was superior in most parameters evaluated. Therefore, according to the results obtained, the association of fluconazole with the ß (1-3) glucan immunomodulator can be a beneficial alternative to a considerable number of cryptococcosis-bearing patients.

A Criptococose é uma enfermidade micótica sistêmica, subaguda ou crônica, que acomete a cavidade nasal, tecidos paranasais e pulmões do homem, animais domésticos e silvestres, podendo disseminar-se para o sistema nervoso central, olhos, pele e outros órgãos. Considerando as dificuldades terapêuticas no tratamento da micose em pequenos animais, incluindo toxicidade, desenvolvimento de resistência aos antifúngicos tradicionalmente utilizados e longos períodos de tratamento da enfermidade, o estudo objetivou avaliar a eficácia do imunomodulador ß (1-3) glucana isoladamente e em associação ao fluconazol no tratamento da criptococose experimental. Foram utilizados 100 camundongos (Mus musculus), cepa UFPel, albinos, os quais foram divididos em cinco grupos de 20 animais. O tratamento dos animais com criptococose experimental foi iniciado sete dias após a inoculação. O grupo Controle (G1) foi tratado com 0,1 ml de água destilada estéril, o grupo Fluconazol (G2) com 5 mg/kg de fluconazol, o grupo Fluconazol associado a ß (1-3) glucana (G3) com 5 mg/kg de fluconazol associado a 0,5 mg de ß (1-3) glucana, o grupo Glucana dose I (G4) com 0,5 mg de ß (1-3) glucana e o grupo Glucana dose II (G5) recebeu 0,25 mg de ß (1-3) glucana. Após acompanhamento clínico durante as seis semanas de tratamento os animais foram eutanasiados e necropsiados para avaliação anatomopatológica dos órgãos, quantificação das unidades formadoras de colônias fúngicas (UFCs), retroisolamento e avaliação histopatológica. Nas avaliações clínicas o G3 foi sempre superior ao G1 e G2, sendo que na última avaliação clínica individual, os animais pertencentes ao G3 não apresentavam nenhum sintoma clínico. O G2 teve um menor número de órgãos afetados e de alterações macroscópicas, seguido pelo G3, sendo que no primeiro não foram observadas lesões em órgãos-alvo, como pulmão e cérebro. O grupo com maior número de lesões e órgãos afetados foi o G1. Nos parâmetros retroisolamento e UFCs, o G3 foi superior aos outros tratamentos, seguido pelo G2, sendo que o pior grupo, ou seja, aquele que teve um maior número de isolamentos e maior quantificação de UFCs foi o G1. Conforme os resultados obtidos os tratamentos G2 e G3 foram os mais eficazes para a remissão da criptococose experimental, no entanto, G3 apresentou uma superioridade na maioria dos parâmetros avaliados. Portanto, de acordo com os resultados obtidos, a associação de fluconazol ao imunomodulador ß (1-3) glucana pode ser uma alternativa benéfica em considerável quantidade de pacientes com criptococose. / Cryptococcosis is a systematic sub acute or chronic mycotic condition that affects the nasal cavity, paranasal tissues and the lungs of humans, as well as domestic and wild animals, which can spread to the central nervous system, skin, and other organs. Considering the therapeutic difficulties in treating this mycosis in small animals, which include toxicity, resistance towards traditionally used antifungals, and the extended treatment of this condition, this study aimed to evaluate the ß (1-3) glucan immunomodulator efficacy when used both alone and in association with fluconazole in the treatment of experimental cryptococcosis. One Hundred albino UFPel strain mice (Mus musculus), divided into five groups of 20 animals each, were used. The treatment of these animals with experimental cryptococcosis was started seven days following inoculation. The control group (G1) was treated with 0,1 ml sterile distilled water. The fluconazole group (G2) was treated with 5 mg/kg fluconazole; the fluconazole associated with ß glucan (1-3) group (G3) was treated with 5 mg/kg fluconazole associated with 0,5 mg ß (1-3) glucan ; the group glucan dose I (G4) was treated with 0,5 mg ß (1-3) glucan, and the group glucan dose II (G5) was administered 0,25 mg ß (1-3) glucan. After the six-week treatment clinical follow-up, the aimals were euthanized and necropsied for anatomopathological evaluation of organs, quantification of fungal colony-forming units (FCUs), retro isolation and histopathological evaluation. In clinical evaluations, G3 was always superior to G1 and G2, and in the last individual clinical evaluation, G3 animals did not show any clinical symptoms. G2 had a smaller number of affected organs and microscopic alterations, followed by G3. In G2, target organ lesions, such as those in the lungs and brain, were not found. Considering retro isolation and FCU parameters, G3 was superior to other treatments, followed by G2; the worst group, that is, the one with the highest isolation occurrence and greatest FCU quantification, was G1. According to the results obtained, treatments G2 and G3 were the most efficient in experimental cryptococcosis remission; however, G3 was superior in most parameters evaluated. Therefore, according to the results obtained, the association of fluconazole with the ß (1-3) glucan immunomodulator can be a beneficial alternative to a considerable number of cryptococcosis-bearing patients.

A Criptococose é uma enfermidade micótica sistêmica, subaguda ou crônica, que acomete a cavidade nasal, tecidos paranasais e pulmões do homem, animais domésticos e silvestres, podendo disseminar-se para o sistema nervoso central, olhos, pele e outros órgãos. Considerando as dificuldades terapêuticas no tratamento da micose em pequenos animais, incluindo toxicidade, desenvolvimento de resistência aos antifúngicos tradicionalmente utilizados e longos períodos de tratamento da enfermidade, o estudo objetivou avaliar a eficácia do imunomodulador ß (1-3) glucana isoladamente e em associação ao fluconazol no tratamento da criptococose experimental. Foram utilizados 100 camundongos (Mus musculus), cepa UFPel, albinos, os quais foram divididos em cinco grupos de 20 animais. O tratamento dos animais com criptococose experimental foi iniciado sete dias após a inoculação. O grupo Controle (G1) foi tratado com 0,1 ml de água destilada estéril, o grupo Fluconazol (G2) com 5 mg/kg de fluconazol, o grupo Fluconazol associado a ß (1-3) glucana (G3) com 5 mg/kg de fluconazol associado a 0,5 mg de ß (1-3) glucana, o grupo Glucana dose I (G4) com 0,5 mg de ß (1-3) glucana e o grupo Glucana dose II (G5) recebeu 0,25 mg de ß (1-3) glucana. Após acompanhamento clínico durante as seis semanas de tratamento os animais foram eutanasiados e necropsiados para avaliação anatomopatológica dos órgãos, quantificação das unidades formadoras de colônias fúngicas (UFCs), retroisolamento e avaliação histopatológica. Nas avaliações clínicas o G3 foi sempre superior ao G1 e G2, sendo que na última avaliação clínica individual, os animais pertencentes ao G3 não apresentavam nenhum sintoma clínico. O G2 teve um menor número de órgãos afetados e de alterações macroscópicas, seguido pelo G3, sendo que no primeiro não foram observadas lesões em órgãos-alvo, como pulmão e cérebro. O grupo com maior número de lesões e órgãos afetados foi o G1. Nos parâmetros retroisolamento e UFCs, o G3 foi superior aos outros tratamentos, seguido pelo G2, sendo que o pior grupo, ou seja, aquele que teve um maior número de isolamentos e maior quantificação de UFCs foi o G1. Conforme os resultados obtidos os tratamentos G2 e G3 foram os mais eficazes para a remissão da criptococose experimental, no entanto, G3 apresentou uma superioridade na maioria dos parâmetros avaliados. Portanto, de acordo com os resultados obtidos, a associação de fluconazol ao imunomodulador ß (1-3) glucana pode ser uma alternativa benéfica em considerável quantidade de pacientes com criptococose. / Cryptococcosis is a systematic sub acute or chronic mycotic condition that affects the nasal cavity, paranasal tissues and the lungs of humans, as well as domestic and wild animals, which can spread to the central nervous system, skin, and other organs. Considering the therapeutic difficulties in treating this mycosis in small animals, which include toxicity, resistance towards traditionally used antifungals, and the extended treatment of this condition, this study aimed to evaluate the ß (1-3) glucan immunomodulator efficacy when used both alone and in association with fluconazole in the treatment of experimental cryptococcosis. One Hundred albino UFPel strain mice (Mus musculus), divided into five groups of 20 animals each, were used. The treatment of these animals with experimental cryptococcosis was started seven days following inoculation. The control group (G1) was treated with 0,1 ml sterile distilled water. The fluconazole group (G2) was treated with 5 mg/kg fluconazole; the fluconazole associated with ß glucan (1-3) group (G3) was treated with 5 mg/kg fluconazole associated with 0,5 mg ß (1-3) glucan ; the group glucan dose I (G4) was treated with 0,5 mg ß (1-3) glucan, and the group glucan dose II (G5) was administered 0,25 mg ß (1-3) glucan. After the six-week treatment clinical follow-up, the aimals were euthanized and necropsied for anatomopathological evaluation of organs, quantification of fungal colony-forming units (FCUs), retro isolation and histopathological evaluation. In clinical evaluations, G3 was always superior to G1 and G2, and in the last individual clinical evaluation, G3 animals did not show any clinical symptoms. G2 had a smaller number of affected organs and microscopic alterations, followed by G3. In G2, target organ lesions, such as those in the lungs and brain, were not found. Considering retro isolation and FCU parameters, G3 was superior to other treatments, followed by G2; the worst group, that is, the one with the highest isolation occurrence and greatest FCU quantification, was G1. According to the results obtained, treatments G2 and G3 were the most efficient in experimental cryptococcosis remission; however, G3 was superior in most parameters evaluated. Therefore, according to the results obtained, the association of fluconazole with the ß (1-3) glucan immunomodulator can be a beneficial alternative to a considerable number of cryptococcosis-bearing patients.

Submitted by Adriana Martinez (amartinez@unoeste.br) on 2017-08-15T19:12:52Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Bianca.pdf: 748434 bytes, checksum: 25858a86b136011052140d31f626de30 (MD5) / Made available in DSpace on 2017-08-15T19:12:52Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Bianca.pdf: 748434 bytes, checksum: 25858a86b136011052140d31f626de30 (MD5) Previous issue date: 2016-11-29 / The purpose of this study was to evaluate the effect of supplementation with L-arginine, in different doses, on body weight, hemogram, production of IgA, IgG and IgM immunoglobulins and in the small bowel inflammatory process of Wistar rats subjected to chemotherapy. A total of 32 Rattus novergicus Wistar rats were randomly distributed in 4 groups (8 rats / group) with similar body weight: control group (CG), the rats were given ration and water; G5-FU was given ration, water and a dose of 5-FU; Garg295 + 5-FU and Garg458 + 5-FU were given, respectively, 295 mg and 458 mg of L-arginine, which was added to the water and a dose of 5-FU. 5-FU was applied after the adaptation period (day 7) in a single dose of 200 mg of 5-FU / kg body weight, intraperitoneally. The rats were weighed individually, always in the morning and without previous fasting, to determine the weight gain before and after the application of 5-FU. Four days after application of 5-FU, the rats were anesthetized with thionembutal, blood and small intestine samples were collected. Supplementation with 295 mg of L-arginine was shown to be beneficial in reducing body weight loss during treatment with the 5-FU chemotherapeutic. The concentration of platelets was lower (P <0.05) in the G5-FU and GArg458 + 5-FU groups, but similar between the GC and GArg295 + 5-FU groups. The total leukocyte concentration showed a significant reduction in the G5-FU, GArg295 + 5-FU and GArg458 + 5-FU groups when compared to the GC group, however, there was a significant reduction of neutrophils only in the G5-FU group. There was an increase (P <0.05) in fibrinogen concentration in the groups treated with L-arginine (GArg295 + 5-FU and GArg458 + 5-FU). In the GArg295 + 5-FU and GArg458 + 5FU groups, the serum IgA concentration increased significantly in relation to the G5-FU group. Rats supplemented with L-arginine had a minor inflammatory process in the small intestine. It was concluded that supplementation with 295 mg of L-arginine / day attenuated the immediate body weight loss of Wistar rats subjected to 5-FU chemotherapy and that daily supplementation with both 295 mg and 458 mg of L-arginine may ease some side effects of 5-FU such as thrombocytopenia, neutropenia and immunomodular production of IgA, in addition to reducing intestinal inflammatory processes. / Este estudo teve o objetivo deste estudo foi avaliar o efeito da suplementação com L-arginina, em diferentes doses, no peso corporal, no hemograma, na produção das imunoglobulinas IgA, IgG e IgM e no processo inflamatório do intestino delgado de ratos Wistar submetidos à quimioterapia. Foram utilizados 32 Rattus novergicus da linhagem Wistar, distribuídos aleatoriamente em 4 grupos (8 ratos/grupo) com peso corporal semelhante: grupo controle (GC), os ratos receberam ração e água; G5-FU receberam ração, água e uma dose de 5-FU; Garg295+5-FU e Garg458+5-FU que receberam ração, 295 mg e 458 mg de L-arginina, respectivamente, adicionada na água e uma dose de 5-FU. A 5-FU foi aplicada após o período de adaptação (dia 7) na dose única de 200 mg de 5-FU/Kg de peso corporal, por via intraperitoneal. Os ratos foram pesados individualmente, sempre no período da manhã e sem jejum prévio, para determinação do ganho de peso antes e depois da aplicação da 5-FU. Quatro dias após da aplicação da 5-FU, os ratos foram anestesiados com tionembutal e colheu-se amostras de sangue e do intestino delgado. A suplementação com 295 mg de L-arginina mostrou-se benéfica na redução de perda de peso corporal durante o tratamento com o quimioterápico 5-FU. A concentração de plaquetas foi menor (P < 0,05) nos grupos G5-FU e GArg458+5-FU, porém, semelhantes entre os grupos GC e GArg295+5-FU. A concentração de leucócitos totais apresentou redução significativa nos grupos G5-FU, GArg295+5-FU e GArg458+5-FU quando comparados ao grupo GC, entretanto, houve redução significativa de neutrófilos somente no grupo G5-FU. Ocorreu um aumento (P < 0,05) na concentração de fibrinogênio nos grupos tratados com L-arginina (GArg295+5-FU e GArg458+5-FU). Nos grupos GArg295+5-FU e GArg458+5FU, a concentração sérica de IgA aumentou significativamente em relação ao grupo G5-FU. Os ratos suplementados com L-arginina apresentaram menor processo inflamatório no intestino delgado. Conclui-se que a suplementação com 295 mg de L-arginina/dia atenuou a perda de peso corporal imediata dos ratos Wistar submetidos à quimioterapia com 5-FU e que a suplementação diária tanto com 295 mg como 458 mg de L-arginina pode amenizar alguns efeitos colaterais da 5-FU como a plaquetopenia, a neutropenia e imunomodular a produção de IgA, além de reduzir processos inflamatórios intestinais.