Search

Made available in DSpace on 2015-06-17T19:34:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-07-30. Added 1 bitstream(s) on 2015-06-18T12:47:14Z : No. of bitstreams: 1 000826959_20150730.pdf: 4030174 bytes, checksum: f38556cf39a6216769a4090df5ef6aaf (MD5) Bitstreams deleted on 2015-08-03T12:21:14Z: 000826959_20150730.pdf,. Added 1 bitstream(s) on 2015-08-03T12:22:26Z : No. of bitstreams: 1 000826959.pdf: 12698179 bytes, checksum: 8cb2268647197309feff08e38a458970 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) A artrite reumatoide (AR) é uma doença autoimune que compromete as articulações. A maioria das terapias utilizadas para o seu tratamento é baseada na inibição global da resposta imune, podendo aumentar a susceptibilidade a agentes infecciosos. O objetivo geral deste projeto foi definir estratégias tolerogênicas específicas para profilaxia ou terapia da AR. Para isso, em uma primeira etapa, estabelecemos um modelo de artrite induzida por proteoglicano (PG) bovino. Camundongos BALB/c fêmeas retired breeders imunizados com PG bovino associado ao adjuvante brometo de dimetil dioctadecil amônio apresentaram os sinais clínicos característicos da artrite, como eritema e edema decorrentes da inflamação articular de uma ou mais patas. A análise histopatológica mostrou a presença de hiperplasia sinovial, infiltrado inflamatório (formação de pannus), destruição da cartilagem e erosão óssea. A incidência da doença foi de 100% e os animais artríticos produziram níveis significativos de citocinas pró e anti-inflamatórias e anticorpos IgG1 e IgG2a anti-PG. Em uma segunda etapa, testamos o potencial profilático do PG. A inoculação de três doses de 50 μg de PG determinou um efeito profilático caracterizado pela diminuição significativa da incidência da artrite e do escore clínico dos animais. A diminuição da produção de IFN-g e IL-17, bem como o aumento da produção de IL-4 e IL-10 por células esplênicas estimuladas com PG, podem estar contribuindo para o efeito profilático observado neste grupo. Em uma terceira etapa, avaliamos o potencial terapêutico da associação da vitamina D ativa (VitD3) com o PG. A associação VitD3+PG determinou um efeito terapêutico na artrite experimental caracterizado por diminuição significativa do escore clínico. Este feito foi confirmado pela análise histopatológica que revelou que a maioria das patas do grupo tratado apresentou estrutura articular bem preservada, ... Rheumatoid arthritis (RA) is an autoimmune disease that affects the joints. Most of the therapies used for RA treatment are based on general suppression of immune response, which may increase the susceptibility to infectious agents. The main objective of this work was to establish specific tolerogenic strategies for prophylaxis or therapy of RA. For this purpose, we first established a model of arthritis induced by bovine proteoglycan (PG). Female BALB/c retired breeder mice immunized with bovine PG associated with dimethyl-dioctadecyl ammonium bromide adjuvant developed a typical arthritis characterized by erythema and edema resulting from joint inflammation of one or more paws. Histopathological analysis showed the presence of synovial membrane hyperplasia, inflammatory infiltrates (pannus formation), cartilage destruction and bone erosion. Disease incidence was 100% and the arthritic mice produced significant levels of pro and anti-inflammatory cytokines and IgG1 and IgG2a anti-PG antibodies. Further, we tested the prophylactic potential of PG. Three doses of 50 μg of PG determined a prophylactic effect characterized by a significant decrease in both, arthritis incidence and clinical scores. The decrease in IFN-g and IL-17, as well as the increase in IL-5 and IL-10 production by spleen cells stimulated with PG may be contributing to the prophylactic effect observed in this group. Lastly, we evaluated the therapeutic potential of the combination of active vitamin D (VitD3) with PG. The VitD3+PG association determined a therapeutic effect in experimental arthritis. There was a significant decrease in the clinical scores after VitD3+PG treatment that was confirmed by the histopathological analysis. Most mice paws from the treated group presented well preserved joint structures that were similar to the ones present in healthy animals. Both pro and anti-inflammatory cytokines were decreased after this treatment. No differences were ...

Made available in DSpace on 2016-02-05T18:29:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-06-11. Added 1 bitstream(s) on 2016-02-05T18:33:59Z : No. of bitstreams: 1 000854046.pdf: 1970431 bytes, checksum: a29a47eeb44d6d6a90258f9d36ebbdf4 (MD5) Uma das aplicações da terapia imunossupressora é evitar que ocorra rejeição em situações de transplante de órgãos e auxiliar na sobrevida dos indivíduos. O ácido micofenólico (A.M.) é um imunossupressor de caráter anti-proliferativo, inibidor da inosina 5-monofosfato desidrogenase, porém, apresenta baixa biodisponibilidade oral e por isso na terapêutica utiliza-se o seu pró-fármaco: micofenolato de mofetila. Este trabalho teve como objetivo a síntese de pró-fármacos mútuos do ácido micofenólico ligados a derivados ftalimídicos a fim de garantir-lhe melhorias farmacocinéticas e farmacodinâmicas. Os derivados ftalimídicos encontrados na estrutura de compostos, por exemplo, na talidomida, utilizada em doenças auto-imunes, agem como imunossupressores por inibição de citocinas pró-inflamatórias. Os pró-fármacos foram obtidos com rendimentos que variaram entre 40-53%. As novas moléculas foram caracterizadas utilizando métodos analíticos como ressonância magnética nuclear (RMN), espectrometria na região de infravermelho e espectrometria de massas. Além disso, o coeficiente de partição (log P) foi determinado pelo método de HPLC e usando os programas de Chem Draw® Ultra e AlogPS®.O log P experimental dos derivados apresentou valores entre 2,29 e 4,09. Avaliou-se a citotoxicidade, liberação óxido nítrico (NO) e de citocinas (IL-1β e TNF-α) usando linhagens celulares de macrófagos murinos. A genotoxicidade in vivo foi avaliada usando o teste de micronúcleo. Todos os compostos apresentaram viabilidade celular superior a 70% nas concentrações usadas. O pró-fármaco (E)-2-(1,3-dioxoisoindolin-2-il) etil6-(4-hidroxi-6-metóxi-7-metil-3-oxo-1,3-dihidroisobenzofuran-5-il)-4-metilhex-4-enoato (3a) apresentou valores de IC50 de 200 μM. Na avaliação da inibição de TNF-α todos os pró-fármacos apresentam atividade nas concentrações utilizadas... One of the applications is immunosuppressive therapy to prevent rejection occurs in situations of organ transplantation and assist in the survival of individuals. Mycophenolic acid (MA) is an immunosuppressive anti -proliferative character inhibitor of inosine 5 -monophosphate dehydrogenase but has a low oral bioavailability and therefore therapeutic uses is the prodrug thereof: mycophenolate mofetil. This work aimed at the synthesis of mutual prodrugs of mycophenolic acid derivatives linked to ftalimidic to ensure you Pharmacokinetic and pharmacodynamic improvements. The ftalimdic derived from compounds found in the structure, for example in thalidomide used in autoimmune diseases, they act as immunosuppressants by inhibiting pro-inflammatory cytokines. The prodrugs were obtained with yields ranging from 40-53 %. The new molecules were characterized using analytical methods such as nuclear magnetic resonance (NMR) spectroscopy in the infrared region and mass spectrometry. In addition, the partition coefficient (log P) was determined by HPLC method using programs Chem Draw Ultra ® and AlogPS ®. Experimental log P of the derivatives showed values between 2.29 and 4.09. Cytotoxicity was assessed, the release nitric oxide (NO) and cytokines (IL- 1β and TNF- α) using murine macrophage cell lines. The in vivo genotoxicity was assessed using the micronucleus test. All compounds showed cell viability above 70 % in the concentrations used. The prodrug (E) -2 - (1,3- dioxoisoindolin -2- yl) etil 6 -(4 -hydroxy- 6-methoxy -7-methyl -3-oxo -1,3- dihydroisobenzofuran -5- yl) 4- methylhex -4- enoate ( 3a ) showed IC50 values of 200 mM . In evaluating the inhibition of TNF- α all prodrugs exhibit activity at the concentrations used were the most active, and 3a (E) - (1,3- dioxoisoindolin -2- yl) methyl 6 - (4 -hydroxy -6- methoxy -7-methyl -3-oxo -1,3- dihydroisobenzofuran -5-yl )-4 -methylhex- 4-enoate (3c) with IC50 values of 18.75 mM . In ...

Made available in DSpace on 2015-04-09T12:28:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-01-21Bitstream added on 2015-04-09T12:48:05Z : No. of bitstreams: 1 000814238.pdf: 438890 bytes, checksum: f7cc8219db858e87f195b5d08dbb8b04 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) itochondrial dysfunction is associated with the manifestation and origin of diseases and disorders. The new paradigm complements the current mitochondrial dogma, whereby molecules present on or inside the mitochondria may act as immune regulators in response to stress or pathogens. Canine distemper virus infection (CDV) is responsible to immunosuppressive stage when the virus replicates among immune cells. For this purpose, canine peripheral blood mononuclear cells (PBMC) collected from healthy dogs were cultured and infected by CDV vaccine strain (Onderstepoort) and after 24 h post-infection (p.i.) superoxide dismutase (SOD1), antioxidant like protein 1 (AOP-1) and heat shock protein 70 (Hsp-70) enzymes were search in PBMC by immunofluorescence. The expression of mRNA of respective genes was performed in infected and uninfected canine PBMC at 24 h post-infection by real time polymerase chain reaction. Mitochondrial dysfunction was evaluated by the use of MitoTracker™Green and JC-1 probes at the same post-infection time. The vaccine strain induced loss of PBMC viability in more than 80% of infected cells in comparison to control group (p