Made available in DSpace on 2014-12-17T14:03:24Z (GMT). No. of bitstreams: 1 josaneRL.pdf: 1324406 bytes, checksum: 22ffd7cc30434eb0cb8d5f0e9616a218 (MD5) Previous issue date: 2001-03-08 / GB virus type C (GBV-C) appears to promote a Th1 response and is associated with prolonged survival in HIV-infected people. L. chagasi causes a spectrum of illness that varies from severe visceral leishmaniasis, a disease that in the majority of cases is fatal if not treated, to self resolution of infection and development of positive DTH response that is protective against symptomatic disease. To determine if GBV-C viremia might influence the outcome of Leishmania infection, we characterized GBV-C status in a cohort of subjects residing in a L. chagasi endemic area in Brazil. GBV-C viremia was more prevalent in blood donors from urban than in periurban regions of Natal, Brazil (16% and 7.5% respectively). Evidence of prior GBV-C (anti-E2 antibodies) was detected in 24% and 12%of these groups respectively. Anti-E2 increased with age (p= 0.0121). No difference in GBV-C viremia was found in the DTH+ and VL groups (p= 0.269); however, subjects with visceral leishmaniasis were more likely to have anti-E2 than DTH+ subjects (p=0.0012), and DTH induration was smaller in subjects with E2 antibodies (4.5 mm) compared those without (7.12 mm) (p= 0.002). Furthermore, the size of the Leishmania DTH response was greater in GBV-C viremica subjects (6.8 mm) compared to non-viremic subjects (3.3 mm; p= 0.0054). There findings suggest that GBV-C virus may promote a type 1 immune response that could influence the outcome of Leishmania infection / GBV-C ? um v?rus membro da fam?lia dos Flavivirus, que tem um aparente papel indutor da resposta Th1 e est? associado ao prolongamento da sobrevida, em pessoas infectadas com HIV. L. chagasi ? respons?vel por um amplo espectro que variam da forma assintom?tica, com resolu??o espont?nea da infec??o e, consecutivamente, desenvolvimento resposta positivo para o DTH; a forma severa da leishmaniose visceral (LV), que na maioria dos casos pode ser fatal se n?o tratada. Para determinar se o GBV-C pode influenciar na resolu??o da infec??o por Leishmania, n?s pesquisamos a preval?ncia de GBV-C em uma popula??o residente em ?rea end?mica para L. chagasi no Brasil. Foi observado que a viremia de GBV-C foi maior em indiv?duos residentes na regi?o urbana, que nos residentes em regi?o periurbana de Natal, Brasil (16% e 7,5%, respectivamente). Evidencia de previa infec??o de GBV-C (anticorpo anti-E2) foi detectada em 24% e 12% nesses grupos respectivamente, aumentando com a idade (p= 0.0121). N?o foi encontrada diferen?a da viremia de GBV-C entre os grupos DTH+ e LV (p= 0.269); Contudo, o grupo dos indiv?duos LV teve mais casos de anti-E2 (p= 0,0012), e a endura??o do DTH foi menor em indiv?duos com anti-E2 comparado aos sem anti-E2 (4.5 mm x 7.12 mm, respectivamente; p= 0,002). O inverso foi observado para os indiv?duos positivos para RNA de GBV-C (6.8 mm) comparados aos sem (3.3 mm; p= 0.0054). Esses achados sugerem que o GBV-C pode promover o tipo 1 de resposta imune e pode com isso influenciar na resolu??o da infec??o por Leishmania

Made available in DSpace on 2014-12-17T14:03:31Z (GMT). No. of bitstreams: 1 JailmaAL.pdf: 4054706 bytes, checksum: f46f1a4accf9845bb01d4688f7c0745c (MD5) Previous issue date: 2009-06-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. They are extracted mainly from the extracellular matrix of brown algae and echinoderms. The brown alga Spatoglossum schr?ederi (Dictyotaceae) has three heterofucans named A, B and C. Our research group have been extracted non anticoagulant heterofucan from S. schr?ederi which possess antithrombotic activity in vivo. However, their toxicity in vitro and in vivo has not yet been determined. For the results in toxicity in vitro, we observed that the fucan A at 20, 500 and 1000 μg/plate showed no mutagenic activity in Kado test (Microsuspension), when the bacterial strains TA97a, TA98, TA100 and TA102, with and without S9 were used. The comet assay showed that fucan A (from 20 to 1000 μg/mL) did not cause any genotoxic effect on CHO cells. There was no damage to the DNA of these cells, as evidenced by the tail length and tail moment, which were similar to that found for the negative control. The fucan A from S. schr?ederi was administered at 20 μg/g of rat (dose which it showed high antithrombotic activity) during two months. After that, the animals were killed and examined. The data showed that fucan A did not cause any change in biochemistry and hematological parameters, as well as, in the morphology and size of the rat s organs analyzed. In conclusion, this study indicates that fucan is a compound with potential pharmacological that has no toxicity / Fucana ? um termo usado para denominar uma fam?lia de polissacar?deos sulfatados ricos em L-fucose. S?o extra?dos principalmente da matriz extracelular de algas marrons e equinodermas. A alga marrom Spatoglossum schr?ederi (Dictyotaceae) possui tr?s heterofucanas nomeadas de fucanas A, B e C. Tem sido proposto o uso de fucanas como alternativas para anticoagulantes. Nosso grupo de pesquisa extraiu uma heterofucana n?o anticoagulante da alga S. schr?ederi que tem uma elevada atividade antitromb?tica in vivo. No entanto, a sua toxicidade in vitro e in vivo ainda n?o foi determinada. Para os resultados obtidos na toxicidade in vitro, observou-se que a fucana A nas concentra??es de 20, 500 e 1000 μg/placa n?o mostram atividade mutag?nica em teste Kado (Microsuspens?o) utilizando a cepas bacterianas TA97a, TA98, TA100 e TA102, com e sem S9. No ensaio do cometa a presen?a da fucana A n?o provocou nenhum efeito genot?xico nas concentra??es testadas de 20, 500 e 1000 μg/mL. N?o houve dano no DNA dessas c?lulas, como evidenciado pelo tail lenght e tail moment, sendo semelhantes ao encontrado para o controle negativo. A fucana A da alga Spatoglossum schr?ederi quando administrada nos animais durante o per?odo de dois meses, n?o provocou altera??o dos par?metros hematol?gicos, bioqu?micos, morfologia e tamanho dos ?rg?os analisados. Esse teste n?o demonstrou que a fucana, na dose que apresenta atividade antitromb?tica, apresenta toxicidade. Os dados do trabalho indicam que esta fucana ? um composto com potencial farmacol?gico que n?o apresenta toxidade, esse fato da seguran?a para que testes futuros com esse pol?mero sejam realizados, inclusive testes em humanos

Made available in DSpace on 2014-12-17T14:03:36Z (GMT). No. of bitstreams: 1 RuthMO_DISSERT_partes_autorizadas.pdf: 1326808 bytes, checksum: b857ffe38a32dadfb7993ebe2bf2b79c (MD5) Previous issue date: 2011-03-21 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The species of the genus Marsdenia, Apocynaceae, are widely used in folk medicine of several countries. In Brazil is found several species belonging to this genus. The in vitro antioxidant, anticoagulant and antiproliferative activities were evaluated to aqueous extracts of stalk, leaf and root of Marsdenia megalantha. In the total antioxidant capacity assay (expressed as ascorbic acid equivalents) the stalk extract showed 76.0 mg/g, while leaf and root extracts 141.3 mg/g and 57.0 mg/g, respectively. The stalk and leaf extracts showed chelating activity around 40% at 1.5 mg/mL, while root extract, at the same concentration showed, 17%. Only the leaf extract showed a significant ability in superoxide scavenging (80% at 0.8 mg/mL). Any extract was able in scavenge hydroxyl, as well anticoagulant activity. The antiproliferative activity of the extracts was evaluated against HeLa tumor cell line. The extracts inhibited in a dose-dependent manner the cell growth. However, the leaf extract showed 80% of inhibition at 1.0 mg/mL, while stalk and root extracts inhibited 63% and 30%, respectively. To assess the mechanism of cell death caused by the leaf extract in HeLa, was performed flow cytometry and western blot. The results show that leaf extract induces cell death by apoptosis through an activation caspase-independent pathway. These data indicate that stalk and leaf extracts obtained have potential to be used as antioxidants and anticancer drugs / As esp?cies do g?nero Marsdenia, Apocynaceae, s?o bastante utilizadas na medicina popular de v?rios pa?ses. No Brasil s?o encontradas v?rias esp?cies pertencentes a esse g?nero. As atividades antioxidante, anticoagulante e antiproliferativa foram avaliadas para os extratos de caule, folha e raiz de Marsdenia megalantha. Na capacidade antioxidante total (expressa como equivalente de ?cido asc?rbico), o extrato do caule apresentou 76,0 mg/g, enquanto os extratos da folha e raiz apresentaram, respectivamente, 14,3 mg/g e 57,0 mg/g. Os extratos do caule e da folha mostraram habilidade quelante em torno de 40% na concentra??o de 1,5 mg/mL, enquanto o extrato da raiz, na mesma concentra??o, apresentou 17%. Apenas o extrato da folha apresentou uma capacidade significante em sequestrar radicais super?xidos (80% em 0,8 mg/mL). Nenhum extrato mostrou capacidade em seq?estrar radicais hidroxila, bem como atividade anticoagulante. A atividade antiproliferativa dos extratos foi avaliada contra a linhagem tumoral HeLa. Os extratos inibiram, de forma dose-dependente, o crescimento celular. Entretanto, o extrato da folha foi capaz de inibir em 80% a prolifera??o celular na concentra??o de 1,0 mg/mL, enquanto os extratos de caule e raiz inibiram 63% e 30%, respectivamente. Para avaliar o mecanismo de morte celular causada pelo extrato da folha nas c?lulas HeLa, foi realizada citometria de fluxo e western blot. Os resultados mostraram que o extrato da folha induz morte celular por apoptose atrav?s de uma via de ativa??o independente de caspase. Estes resultados indicam que os extratos de caule e folha obtidos t?m potencial para serem futuramente utilizados como f?rmacos antioxidantes e antic?ncer

Made available in DSpace on 2014-12-17T14:03:39Z (GMT). No. of bitstreams: 1 CristinaIO_DISSERT.pdf: 1652503 bytes, checksum: 5827ebf97f52c6102acf9da751e47460 (MD5) Previous issue date: 2012-12-03 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / American visceral leishmaniasis (AVL), caused by Leishmania infantum chagasi (L.i.chagasi), stands as a public health problem in Brazil, with human and canine cases related in all states..Lipid metabolism can be modified in several status of infection. For example, experimental studies show that the cholesterol is necessary to internalization and replication of L.i.chagasi in macrophages through caveolar domains. Patients with AVL present low levels of cholesterol and a visible triglycerides increase. This work aimed to evaluate the lipid metabolism in several post-infection status by L.i.chagasi, including individuals with symptomatic infection (AVL), and asymptomatic. The levels of cholesterol, triglycerides, HDL and reactive C protein, were measured. Individuals with AVL were compared with individuals with assymptomatic infection and presented low levels of total cholesterol (128 ? 6.180 mg/dL vs. 158 ?5.733 mg/dL, p=0.0001), HDL (29 ? 1.746 mg/dL vs. 37 ? 1.647 mg/dL, p=0.0001), increased levels of triglycerides (149.5 mg/dL ? 12.72 vs. 78.00 ? 10.43 mg/dL, p=0.0095) and higher levels of reactive C protein (1.750? 0.4939 mg/dL vs. 0.40 ? 0.1707 mg/dL; p=0.0001). The expression of genes related to lipid metabolism, such as LXR-a, LXR-b, PPAR-a, PPAR-d, PPAR-g and APOE was evaluated by real time PCR. A reduction in the expression of those genes was found in the group of AVL patients corroborating the serum levels of the metabolites earlier quantified. Our findings suggest a modulation of metabolism of lipids, in the chronic phase of AVL, this could facilitate the survival of leishmania, due to the known reduction on the ability of macrophages in presenting antigens efficiently to the T cells due to the reduction in the cholesterol available, it results in a subversion of the host immunity. / A leishmaniose visceral americana (LVA), causada pela Leishmania infantum chagasi (L.i.chagasi) permanece como um problema de sa?de p?blica no Brasil. A LVA cursa com altera??es bioqu?micas e hematol?gicas que podem resultar em caquexia e predisposi??o a infec??es secund?rias. O metabolismo lip?dico pode ser alterado em resposta ? infec??o, sendo que alguns microorganismos podem induzir estas altera??es.. Estudos experimentais mostram que o colesterol ? necess?rio para a internaliza??o e replica??o de L.i.chagasi em macr?fagos. Este trabalho teve como objetivo avaliar se o metabolismo lip?dico sofre altera??es em diversos status p?s-infec??o por L.i.chagasi. Os n?veis de colesterol, triglicer?deos, HDL, prote?na C reativa foram quantificados. A express?o de genes relacionados ao metabolismo lip?dico, como LXR-a, LXR-b, PPAR-a, PPAR-d, PPAR-g e APOE foi determinada por PCR em tempo real. Indiv?duos com LVA quando comparados com indiv?duos com infec??o assintom?tica apresentaram baixos n?veis de coleste rol total (128 ? 6,180 mg/dL vs. 158 ?5,733 mg/dL, p=0,0001), HDL (29 ? 1,746 mg/dL vs. 37 ? 1,647 mg/dL, p=0,001); eleva??o nos n?veis de triglicer?deos (149,5 mg/dL ? 12,72 vs. 78,00 ? 10,43 mg/dL, p=0,0095) e de prote?na C reativa (1,750? 0,4939 mg/dL vs. 0,40 ? 0,1707 mg/dL; p=0,0001). A express?o de genes envolvidos no metabolismo de lip?dos apresentou diminui??es de 3,5 vezes em pacientes com LVA, no caso de PPAR-g. Para outros genes, como PPAR-a, esta diminui??o de express?o chegou a 27,70 vezes para pacientes com LVA em rela??o ao grupo controle. Estes achados podem explicar os n?veis s?ricos alterados de lip?dios para os indiv?duos com LVA. Estes resultados sugerem que a cronicidade da infec??o por Leishmania resulta em uma modula??o do metabolismo de lip?dios, com inibi??o da s?ntese de colesterol. Isso pode facilitar a sobreviv?ncia da leishmania, por potencialmente resultar em redu??o da habilidade dos macr?fagos em apresentar ant?genos eficientemente para c?lulas T frente ? redu??o de colesterol dispon?vel, e isso resulta em uma subvers?o da imunidade do hospedeiro.

Orientador: José Jurandir Fagliari / Coorientador: Daniela Gomes da Silva / Banca: Roberto Calderon Gonçalves / Banca: Julio Augusto Naylor Lisboa / Banca: Mario Roberto Hatayde / Banca: Fernando Antônio de Ávila / Resumo: O presente estudo teve como objetivo avaliar as alterações clínicas e laboratoriais de bezerros bubalinos infectados experimentalmente com Salmonella Dublin e verificar efeito do tratamento com antibiótico florfenicol. Também teve como objetivo avaliar a eficiência da reação em cadeia da polimerase (PCR) comparativamente ao exame bacteriológico no diagnóstico da salmonelose. Para isso, foram utilizados três grupos experimentais (n=6): controle (grupo 1), que receberam, por via oral, 10 ml de caldo BHI; sem tratamento (grupo 2) e tratados com florfenicol (grupo 3), que receberam, por via oral, 108 UFC de S. Dublin suspensas em 10 ml de BHI. Os bezerros foram submetidos ao exame físico antes da inoculação e a cada 24 horas até o sétimo dia após infecção. Diariamente, foram colhidas amostras de sangue para a realização de hemograma e testes bioquímicos (inclusive proteinograma e hemogasometria) e colhidos suabes retais para identificação de S. Dublin por exame bacteriológico e PCR. A infecção experimental induziu hipertermia e diarreia. A inoculação também provocou leucopenia e neutropenia, hipofosfatemia, aumento das concentrações das proteínas fibrinogênio, ceruloplasmina e haptoglobina e diminuição das concentrações de ferro. Os bezerros tratados não apresentaram cura clínica e bacteriológica concluindo-se que o florfenicol não foi efetivo no combate à S. Dublin. A PCR mostrou-se superior ao isolamento bacteriológico na detecção de S. Dublin. Adicionalmente, a eliminação da bactéria nas fezes dos animais infectados após o período de sintomatologia confirma o estado de portador desses animais / Abstract: The aim of the study was to evaluate clinical and laboratorial changes in experimentally Salmonella Dublin-infected buffalo calves and the effect of florfenicol antibiotic treatment. Also, the aim of the study was to compare the performance of standard microbiological techniques and polymerase chain reaction (PCR) for the diagnosis of S. Dublin. Three experimental groups, with six calves, were formed: control group (G1), that orraly received 10 ml of BHI broth; without any treatment group (G2) and treated with florfenicol group (G3), that orally received 108 CFU of S. Dublin suspended in 10 ml of BHI broth. All calves were submitted to physical examination before experimental inoculation and at every 24 hours up to the 7th day after infection. Daily blood samples were collected for hematological and biochemical analysis (including proteinogram and hemogasometry). Daily rectal swabs were collected for the isolation of S. Dublin by standard microbiological techniques and PCR. The experimental infection induced hyperthermia and diarrhea. The inoculation also caused leukopenia and neutropenia, hypophosphatemia, increase of the fibrinogen, ceruloplasmin and haptoglobin concentrations and decrease of the iron concentrations. Calves treated showed no clinical and bacteriological cure, concluding that florfenicol was not effective against S. Dublin. PCR had better performance in the isolation of S. Dublin when compared with standard microbiological techniques. Additionally, elimination of the bacteria in the feces of infected animals after a period of symptoms confirms the carrier status of buffalo calves / Doutor

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-03-03T19:55:15Z No. of bitstreams: 1 ErikaCristinaPinheiroDeCastro_DISSERT.pdf: 2706231 bytes, checksum: fd12403d1b13d2a828b00af3b56d6cc1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-03-07T21:59:52Z (GMT) No. of bitstreams: 1 ErikaCristinaPinheiroDeCastro_DISSERT.pdf: 2706231 bytes, checksum: fd12403d1b13d2a828b00af3b56d6cc1 (MD5) / Made available in DSpace on 2016-03-07T21:59:52Z (GMT). No. of bitstreams: 1 ErikaCristinaPinheiroDeCastro_DISSERT.pdf: 2706231 bytes, checksum: fd12403d1b13d2a828b00af3b56d6cc1 (MD5) Previous issue date: 2013-12-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / O consumo mundial de ?leos vegetais aumentou nos ?ltimos anos devido sua aplica??o na ind?stria aliment?cia, qu?mica, farmac?utica e, recentemente energ?tica. Todavia, as sementes oleaginosas das quais esses ?leos s?o extra?dos envelhecem rapidamente, comprometendo o cultivo e a produtividade destas. O objetivo deste trabalho foi avaliar o efeito do envelhecimento sobre a coordena??o dos sistemas antioxidantes da catalase (CAT) e da ascorbato peroxidase (APX) em c?rtamo e girassol. Para tanto, sementes foram submetidas a envelhecimento acelerado por 3, 6 e 9 dias e cultivadas em papel-toalha umedecido durante 72h. Adicionalmente, antes de serem submetidas ao envelhecimento acelerado, sementes de girassol foram pr?- tratadas por osmopriming com 10 mM de ascorbato (ASC) ou 3 amino 1,2,4 triazole(3-AT), inibidor espec?fico da CAT. O m?todo de envelhecimento artificial utilizado foi eficiente para ambas esp?cies, pois promoveu diminui??o na germina??o, no desenvolvimento de pl?ntulas e no crescimento destas, principalmente no c?rtamo. O envelhecimento provocou inibi??o da atividade CAT para ambas as esp?cies e para compensar tal inibi??o, o girassol aumentou a express?o do mRNA desta enzima, enquanto o c?rtamo mobilizou mais a atividade da APX. A an?lise da express?o da malato sintase e do conte?do de a?ucares demonstrou que o girassol consome suas reservas lip?dicas no estado quiescente, enquanto o c?rtamo ? mais dependente dos carboidratos. O pr?-tratamento com 3-AT inibiu a atividade CAT e estimulou a da APX, enquanto o ASC atuou de forma inversa nesses sistemas. Nenhum dos tratamentos recuperou o decl?nio fisiol?gico decorrente do envelhecimento. Conclui-se que o envelhecimento alterou significativamente o metabolismo antioxidante das oleaginosas e, apesar das varia??es interespec?ficas na resposta a esse processo, a deple??o do sistema antioxidante da CAT foi comum, deforma que a mensura??o da atividade desta enzima pode ser utilizada para identifica??o de lotes de sementes envelhecidas. / World consumption of vegetable oils has increased in recent years because of its application in food, chemical, pharmaceutical and, more recently, energy industry. However, oilseeds, which these oils are extracted, have low viability, affecting the cultivation and productivity of these species. The aim of this study was to analyze the effect of aging on the coordination of catalase (CAT) and ascorbate peroxidase (APX) antioxidant systems in safflower and sunflower. . Therefore, seeds were subjected to accelerated aging for 3, 6 and 9 days and grown in moistened paper towel for 72 hours. Additionally, before accelerated aging, sunflower seeds were pretreated by osmopriming with 10 mM ascorbate (ASC) or 3 amino 1,2,4 triazol (3-AT), a specific inhibitor of CAT activitie. The method of artificial aging used was efficient in both species, because it caused a decrease in germination, seedling development and growth, especially in safflower. The aging caused inhibition of CAT activity for both species and to compensate for such inhibition , sunflower increased mRNA expression of this enzyme , while safflower mobilized over the activity of APX. Analysis of the expression of malate synthase and sugar content demonstrated that sunflower seeds consumes lipid reserves in quiescent state, while the safflower is more dependent on carbohydrate. Pretreatment with 3-AT inhibited CAT activity and stimulated the APX, though with ASC acted reverse on these systems. None of the treatments recovered the physiological decline aging. It is concluded that aging change the oilseeds antioxidant metabolism, despite interspecies variations in response to this process, the depletion of the CAT antioxidant system was common. Because of this we propose that the measurement of CAT activity can be used to identify aging seed lots.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-03-03T19:55:15Z No. of bitstreams: 1 RayssaKarlaDeMedeirosOliveira_DISSERT.pdf: 5161047 bytes, checksum: b9cdb0f408571e0784106130b004445a (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-03-07T22:14:58Z (GMT) No. of bitstreams: 1 RayssaKarlaDeMedeirosOliveira_DISSERT.pdf: 5161047 bytes, checksum: b9cdb0f408571e0784106130b004445a (MD5) / Made available in DSpace on 2016-03-07T22:14:58Z (GMT). No. of bitstreams: 1 RayssaKarlaDeMedeirosOliveira_DISSERT.pdf: 5161047 bytes, checksum: b9cdb0f408571e0784106130b004445a (MD5) Previous issue date: 2014-08-21 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Prote?nas do reparo por excis?o de bases (BER) t?m sido associadas a fun??es al?m do reparo e DNA. A apur?nica/apirimidinica endonuclease 1 (APE1) ? uma prote?na multifuncional envolvida em diversas atividades celulares como ativa??o redox de fatores de transcri??o, processamento de RNA e reparo de DNA. Alguns trabalhos t?m descrito a a??o da prote?na 8-oxoguanina (OGG1) na corre??o de les?es oxidadas no promotor como passo para a transcri??o de citocinas pro-inflamat?rias. Apesar de ser notadamente importante na ativa??o redox de fatores de transcri??o, como o fator nuclear ?B (NF- ?B) e AP-1, a atividade de reparo de APE1 ainda n?o foi associada ? resposta inflamat?ria. Neste trabalho, foram utilizadas an?lises bioinform?ticas e abordagens experimentais para investigar a rela??o entre a inibi??o do reparo de s?tios ab?sicos no DNA pela MX, mol?cula sint?tica inibidora indireta da atividade de reparo de APE1, e a modula??o de resposta inflamat?ria. Os resultados demonstraram que o tratamento de mon?citos com lipopolissacar?deo (LPS) e MX reduziu a express?o de citocinas, quimiocinas e receptores toll-like, e regulou negativamente processos biol?gicos da imunidade, como ativa??o de macr?fagos, e as vias ativadas pelo (NF-?B), fator de necrose tumoral (TNF-?) e interferon, sem induzir morte celular. A an?lise transcript?mica sugere que o tratamento LPS/MX induz disfun??es mitocondriais, estresse de ret?culo endoplasm?tico e ativa??o de vias de autofagia, provavelmente ativadas pelo comprometimento da energ?tica celular e/ou pelo ac?mulo de danos ao DNA, nuclear e mitocondrial. Adicionalmente, prop?e-se que a atividade de reparo de APE1 ? requerida para a transcri??o de genes inflamat?rios pela intera??o com s?tios ab?sicos no promotores espec?ficos e recrutamento de complexos transcricionais durante a sinaliza??o inflamat?ria. Este trabalho apresenta uma nova perspectiva acerca das intera??es entre a atividade do BER e a modula??o de resposta inflamat?ria, e sugere uma nova atividade para a prote?na APE1 como modular da resposta imune de maneira redox-independente. / Base excision repair (BER) proteins has been associated with functions beyond DNA repair. Apurynic/apyrimidinic endonuclease 1 (APE1) is a multifunctional protein involved in a plethora of cellular activities, such as redox activation of transcription factors, RNA processing and DNA repair. Some studies have described the action of the protein 8-oxoguanine (OGG1) in correcting oxidized lesions in promoters as a step in the transcription of pro-inflammatory cytokines. Despite being especially important in redox activation of transcription factors such as nuclear factor ?B (NF-?B) and AP- 1, the repair activity of APE1 has not yet been associated with the inflammatory response. In this study, experimental and bioinformatic analysis approaches have been used to investigate the relationship between inhibition of the repair of abasic sites in DNA by MX, a synthetic molecule designed to inhibt the repair activity of APE1, and the modulation of the inflammatory response. The results showed that treatment of monocytes with lipopolysaccharide (LPS) and MX reduced the expression of cytokines, chemokines and toll-like receptors, and negatively regulated biological immune processes, as macrophages activation, and NF-?B and tumor necrosis factor (TNF-?) and interferon pathways, without inducing cell death. The transcriptomic analysis suggests that LPS/MX treatment induces mitochondrial dysfunction, endoplasmic reticulum stress and activation of autophagy pathways, probably activated by impairment of cellular energy and/or the accumulation of nuclear and mitochondria DNA damage. Additionally, it is proposed that the repair activity of APE1 is required for transcription of inflammatory genes by interaction with abasic sites at specific promoters and recruitment of transcriptional complexes during inflammatory signaling. This work presents a new perspective on the interactions between the BER activity and the modulation of inflammatory response, and suggests a new activity for APE1 protein as modulator of the immune response in a redox-independent manner.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:12Z No. of bitstreams: 1 EvellynCamaraGrilo_DISSERT.pdf: 1083379 bytes, checksum: 5c4378ae79f8da0ab5335caca60a8c5d (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T21:55:39Z (GMT) No. of bitstreams: 1 EvellynCamaraGrilo_DISSERT.pdf: 1083379 bytes, checksum: 5c4378ae79f8da0ab5335caca60a8c5d (MD5) / Made available in DSpace on 2016-04-28T21:55:39Z (GMT). No. of bitstreams: 1 EvellynCamaraGrilo_DISSERT.pdf: 1083379 bytes, checksum: 5c4378ae79f8da0ab5335caca60a8c5d (MD5) Previous issue date: 2015-07-14 / As vitaminas A e E s?o nutrientes que possuem natureza lipof?lica e atuam em v?rios processos biol?gicos importantes, como a imunidade, reprodu??o, crescimento e desenvolvimento. Estas vitaminas s?o essenciais na fase inicial da vida e devem ser transferidas adequadamente da m?e para o filho durante a gesta??o e a lacta??o. A suplementa??o materna com vitamina A ? uma das estrat?gias de controle de sua defici?ncia no grupo materno-infantil, entretanto, estudos com animais evidenciaram que a suplementa??o com altas doses de vitamina A reduziu os n?veis de alfa-tocoferol (vitamina E) no soro e no leite. Assim, o objetivo deste trabalho foi avaliar a influ?ncia da suplementa??o materna com vitamina A sobre a concentra??o de retinol e alfa-tocoferol nos leites colostro e maduro de lactantes. Pu?rperas a termo e saud?veis foram aleatoriamente distribu?das nos grupos controle (n = 44) e suplementado (n = 44). Amostras de sangue e leite colostro foram coletadas no p?s-parto imediato e uma amostra de leite maduro foi coletada ap?s 30 dias. O grupo suplementado recebeu uma suplementa??o com palmitato de retinila (200.000 UI), imediatamente ap?s a primeira coleta de colostro. O retinol e o alfa-tocoferol das amostras biol?gicas foram analisados por Cromatografia L?quida de Alta Efici?ncia (CLAE). Valores s?ricos abaixo de 20 ?g/dL para a vitamina A e 516 ?g/dL para a vitamina E foram indicativos de defici?ncia. As concentra??es de retinol e alfa-tocoferol no soro das lactantes foram 46,4 ? 15,9 ?g/ dL e 1.023,6 ? 380,4 ?g/ dL, respectivamente, sendo consideradas adequadas. No grupo suplementado, verificou-se um aumento significativo dos n?veis de retinol no leite colostro, 24 horas ap?s interven??o (p<0,001), entretanto, n?o foi observada diferen?a estat?stica entre a concentra??o de retinol no leite maduro dos grupos avaliados (p>0,05). Al?m disso, ap?s a suplementa??o materna com vitamina A, houve uma redu??o significativa na concentra??o de alfa-tocoferol no leite colostro (p<0,05), que correspondeu a um decl?nio de 16,4% dos n?veis de vitamina E. Por outro lado, a administra??o do suplemento n?o influenciou os n?veis de alfa-tocoferol no leite maduro (p>0,05). Diante disso, conclui-se que a suplementa??o materna com altas doses de vitamina A aumentou os n?veis desse micronutriente no leite colostro, por?m, reduziu a biodisponibilidade do alfa-tocoferol, o que pode trazer preju?zos ? sa?de do neonato, que possui reservas limitadas de vitamina E ao nascimento. / Vitamins A and E are lipophilic nutrients that act in many important biological processes, such as immunity, reproduction, growth, and development. These vitamins are essential during the initial phase of life and should be properly transferred from mother to child during pregnancy and lactation. Maternal supplementation with vitamin A is one of the strategies for controlling its deficiency in the mother-child dyad, but studies with animals showed that supplementation with high doses of vitamin A reduces the levels of alpha-tocopherol (vitamin E) in the mother?s serum and milk. Hence, the objective of this study was to assess the influence of maternal supplementation with vitamin A on the concentration of retinol and alpha-tocopherol in colostrum and mature milk. Healthy puerperal women with term deliveries were randomly distributed into a control group (n=44) and a supplemented group (n=44). Blood and colostrum samples were collected immediately after delivery, and mature blood samples were collected 30 days later. The supplemented group received 200,000 IU of retinyl palmitate immediately after the first colostrum collection. The retinol and alpha-tocopherol levels in the biological samples were determined by high-performance liquid chromatography (HPLC). Serum vitamin A levels below 20 ?g/dL and serum vitamin E levels below 516 ?g/dL indicated deficiency. The retinol and alpha-tocopherol levels in the maternal serum were considered adequate at 46.4 ? 15.9 ?g/ dL and 1,023.6 ? 380.4 ?g/ dL, respectively. The colostrum retinol levels of the supplemented group increased significantly 24 hours after the intervention (p<0.001). However, the retinol levels in the mature milk of both groups did not differ (p>0.05). Moreover, after maternal supplementation with vitamin A, colostrum alpha-tocopherol level decreased by 16.4%, a significant reduction (p<0.05). However, vitamin A supplementation did not affect the alpha-tocopherol level of mature milk (p>0.05). In conclusion, maternal supplementation with high doses of vitamin A increased the colostrum level of this nutrient but reduced the bioavailability of alpha-tocopherol, which may harm the newborn?s health since newborns have limited vitamin E reserves.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:13Z No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T22:23:31Z (GMT) No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) / Made available in DSpace on 2016-04-28T22:23:31Z (GMT). No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) Previous issue date: 2015-08-07 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / A produ??o de nanoparticulas de prata com extratos naturais tem sido apontada como uma excelente alternativa para potencializar ou fornecer novas aplicabilidades aos extratos. Extratos de polissacar?deos sulfatados de algas (ASP) apresentam propriedades farmacol?gicas, por?m h? poucos relatos da produ??o de nanopart?culas de prata com extratos ricos em polissacar?deos sulfatados (SPN). Assim, neste trabalho sintetizou-se SPN de algas encontradas no Brasil: Spatoglossum schr?ederi, Dictyopteris justii, Sargassum filipendula e Dictyota mertensii. A obten??o dos extratos ricos em polissacar?deos ocorreu por prote?lise seguida por precipita??o com metanol. A s?ntese das diferentes nanopart?culas ocorreu com a adi??o de solu??es de prata 1 mM em solu??es dos diferentes polissacar?deos e mantidos em repouso. Posteriormente as amostras foram centrifugadas e liofilizadas. A forma??o SPN foi confirmada por espectroscopia UV/vis?vel, microscopia eletr?nica de varredura e microscopia de for?a at?mica. O tamanho das SPN foi de 108 ? 2 nm; 82 ? 1nm; 288 ? 52 nm; 104 ? 2 nm para S. schr?ederi; D. justii; S. filipendula; D. mertensii, respectivamente e se manteve est?vel por catorze meses, os potenciais zeta foram negativos e as formas das SPN foram esf?ricas. Os resultados de diversos testes in vitro mostraram que as SPN potencializam as atividades antioxidantes de ASP. As SPN tamb?m foram biocompat?veis com c?lulas normais 3T3 (fibroblastos murinicos). Por outro lado, SPN de S schr?ederi e de D. mertensii tiveram atividade citot?xica (~60%) frente as c?lulas de melanoma mur?nico (B16F10) e acredita-se que a maior atividade citot?xica destas SPN ocorram devido aos seus pequenos tamanhos. SPN tamb?m tiveram grande capacidade antibacteriana. Nanopart?culas de D. justii e S. filipendula tiveram os melhores resultados, sendo necess?rio somente 50 ?g/mL para a morte da bact?ria E. coli e 100 ?g/mL para a morte de S. aureus. Hipotet?za-se que esta atividade ocorra por libera??o de prata pelas SPN. SPN tamb?m foram capazes de induzir a produ??o de ?xido n?trico e citocinas com perfil semelhante a os seus respectivos ASP, com exce??o para SPN de S. schr?ederi. Em geral, os resultados revelaram que a s?ntese de SPN a partir das ASP potencializa efeitos antioxidantes, citot?xico e antibacteriano dos ASP, al?m de apresentar o mesmo efeito imunomodulador de seus respectivos ASP. Os dados obtidos levam a propor que a s?ntese de SPN constituiu-se como um poss?vel mecanismo potencializador de atividades biol?gicas. / The production of silver nanoparticle containing natural extracts has been identified as an excellent alternative to enhance or provide new applicability for these extracts. Sulfated polysaccharides-rich extracts (ASP) from seaweeds showed several pharmacological properties, but there are few reports of the production of silver nanoparticle with sulfated polysaccharides (SPN). Therefore, this paper we synthesized SPN from seaweeds founded in Brazil: Spatoglossum schr?ederi, Dictyopteris justii, Sargassum filipendula e Dictyota mertensii. The obtainment of polysaccharides rich extracts it occurred by proteolysis followed by methanol precipitation. The synthesis of various nanoparticles occurred with the addition of silver solution 1 mM on different polysaccharides solutions and kept at rest. Subsequently the samples were centrifuge and lyophilized. The SPN formation was confirmed by UV/ visible spectroscopy, scanning electron microscopy and atomic force microscopy. The size of SPN was 108 ? 2 nm; 82 ? 1nm; 288 ? 52 nm; 104 ? 2 nm for S. schr?ederi; D. justii; S. filipendula; D. mertensii, respectively. The nanoparticules remained stable for fourteen months, the Zeta potential were negative and forms of SPN were rounded. Data from various antioxidant in vitro tests showed that the SPN were more effective than ASP. The SPN were also biocompatible with the normal cells 3T3 (murine fibroblast). Moreover, SPN of S. schr?ederi and of D. mertensii had cytotoxic activity (~60%) against melanoma cells (B16F10). We believe that the increased of cytotoxic activity of these SPN occur due to their small size. SPN also had great antibacterial capacity. D. justii and S. filipendula nanoparticles had the best results, requiring only 50 mg/mL and 100 mg/mL for induces E. coli and S. aureus death. We hypothesized that this activity occurs by release of SPN?s silver. SPN also increase the nitric oxide and cytokines release from macrophages as similar profile to the respective ASP, except for S. schr?ederi SPN. In general, the results showed that SPN were more effective as antioxidant, antibacterial and cytotoxic agents than their respective ASP, as well as, SPN have shown the same immunomodulation effect of the respective ASP. The results lead to propose that the SPN synthesis constituted as a possible potentiating mechanism of biological activities of ASPs.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:13Z No. of bitstreams: 1 MarinaSampaioDeMenezesCruz_DISSERT.pdf: 1274699 bytes, checksum: f39afb06729aedccb850680b592f9def (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T22:46:18Z (GMT) No. of bitstreams: 1 MarinaSampaioDeMenezesCruz_DISSERT.pdf: 1274699 bytes, checksum: f39afb06729aedccb850680b592f9def (MD5) / Made available in DSpace on 2016-04-28T22:46:18Z (GMT). No. of bitstreams: 1 MarinaSampaioDeMenezesCruz_DISSERT.pdf: 1274699 bytes, checksum: f39afb06729aedccb850680b592f9def (MD5) Previous issue date: 2015-07-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Os produtos naturais derivados de plantas servem de mat?ria-prima para a s?ntese de fitoter?picos, sendo amplamente utilizados por v?rias popula??es em todo o mundo. Dentre estes compostos, destacam-se as saponinas e seus metab?litos. Seu intenso uso ? decorrente de suas propriedades bioativas, somado a sua similaridade estrutural a horm?nios. As saponinas derivam-se em sapogeninas, como, por exemplo, a Diosgenina e a Hecogenina. Al?m de serem utilizadas na ind?stria farmac?utica para a produ??o de horm?nios e anti-inflamat?rios esteroidais, estas duas sapogeninas tamb?m s?o empregadas como fitoter?picos, estando presentes tamb?m em vegetais que comp?em a dieta de muitas popula??es. Apesar da vasta utiliza??o destes compostos, existe uma falta de informa??es sobre a toxicidade gen?tica, o que torna o consumo destas sapogeninas um risco para as popula??es. Diante deste contexto, o presente trabalho apresentou como objetivo investigar as potencialidades t?xicas, em n?vel celular e de DNA, da Diosgenina e da Hecogenina. Ensaios em c?lulas HepG2 para a avalia??o da citotoxicidade, genotoxicidade, e mutagenicidade foram realizados. Considerando os resultados obtidos com a Diosgenina, esta sapogenina revelou efeito citot?xico em concentra??es acima de 30 ?M, assim como induziu significativamente incrementos na frequ?ncia de danos no DNA e de micron?cleos. Por outro lado, a Hecogenina somente se revelou citot?xica acima de 150 ?M, por?m em concentra??es abaixo deste valor, foi capaz de induzir danos ao material gen?tico sem promover um aumento na frequ?ncia de micron?cleos. A an?lise do conjunto destes dados sugere que o consumo de alimentos, fitoter?picos ou de medicamentos que contenham a Diosgenina na sua composi??o apresenta um risco maior de toxicidade, quando comparados com as formula??es a base de Hecogenina, uma vez que a Diosgenina apresenta um potencial mutag?nico demonstrado por meio de um aumento significativo na frequ?ncia de micron?cleos que s?o biomarcadores de instabilidade gen?mica. / Natural products derived from plants are used as raw material for the synthesis of phytotherapeutics, and are widely used by many populations around the world. Among these compounds, there are the saponins and its metabolites. Its intense use is due to its bioactive properties, added to its structural similarity to hormones. Saponins derived in sapogenins, as Diosgenin and Hecogenin. Besides its use by pharmaceutical industries to synthesis of steroidal hormones and anti-inflammatories, these two sapogenins are also used as phytotherapeutics, being also present in vegetables from diets of many populations. Despite the widespread use of these compounds, there are a lack of information about their genetic toxicity, which makes the use of these sapogenins a risk. Given this context, the present work aimed to investigate the toxic potential of Diosgenin and Hecogenin, at cellular and DNA level. Assays with HepG2 cells was performed to evaluate the cytotoxicity, genotoxicity and mutagenicity. Considering the results obtained with Diosgenin, that sapogenin showed cytotoxic effects at concentrations higher than 30 ?M, as well as significantly induced increases in frequency of DNA damage and micronucleus. On the other hand, Hecogenin only revealed cytotoxicity above 150 ?M, however, at concentrations below this value, it was able to induce DNA damage without promote an increase in micronucleus frequency. The analysis of these results together suggests that the consumption of foods, phyotheraputics or medicines containing Diosgenin on its composition, presents a higher risk of toxicity when compared with formulations with Hecogenin, since Diosgenin shows a mutagenic potential demonstrated by a significant increase in micronucleus frequency, which are biomarkers of genome instability.