Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-04-28T12:59:06Z No. of bitstreams: 1 467697.pdf: 3365118 bytes, checksum: f6826fc894e0d18084119ccb17fc33a1 (MD5) / Made available in DSpace on 2015-04-28T12:59:06Z (GMT). No. of bitstreams: 1 467697.pdf: 3365118 bytes, checksum: f6826fc894e0d18084119ccb17fc33a1 (MD5) Previous issue date: 2015-02-27 / Conselho Nacional de Pesquisa e Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The first line of defense of organism is made by phagocytic cells such as neutrophils. Apoptosis and NETosis of neutrophils are two major mechanisms of programmed cell death that differ in their morphological characteristics and effects on the immune system. Apoptosis is characterized by nuclear chromatin packaging and nuclear fragments and this death can be delayed by the presence of pathogens or chemicals components such as lipopolysaccharide (LPS). Neutrophils have other antimicrobial strategy, called neutrophil extracellular traps (NETs), which contributes to the elimination and control of the pathogen. NETosis is induced by infection, inflammation or trauma and represents an innate immune activation mechanism. The objective of this study was to evaluate the effect of Gallic acid (GA) in the control of apoptosis and release NETs. The results show that GA decreased the anti-apoptotic effect of LPS, blocked the induction of NETs and prevented the formation of free radicals induced by LPS. These findings demonstrate that the GA is a novel therapeutic agent for decreasing the exacerbated response of the body against an infectious agent. / A primeira linha de defesa do organismo ? feita por c?lulas fagoc?ticas como os neutr?filos. Apoptose e NETose dos neutr?filos s?o os dois maiores mecanismos de morte celular programada, que diferem em suas caracter?scas morfol?gicas e em seus efeitos sobre o sistema imune. A apoptose ? caracterizada pelo empacotamento da cromatina e dos fragmentos nucleares, por?m esta morte por ser atrasada pela presen?a de pat?genos ou pela presen?a de componentes qu?micos, como o lipopolissacar?deo (LPS). Os neutr?filos possuem outra estrat?gia antimicrobiana, chamada armadilhas extracelulares (NETs), que contribuem para elimina??o e controle do pat?geno. A NETose ? induzida por infec??o, inflama??o ou trauma e, representa um mecanismo de ativa??o da resposta imune inata. O objetivo deste estudo foi avaliar o efeito do ?cido g?lico (AG) no controle da apoptose e forma??o de NETs de neutr?filos. Os reultados mostram que o AG diminuiu o efeito anti-apopt?tico do LPS, bloqueou a libera??o de NETs e preveniu a forma??o de radicais livres induzidos por LPS. Estes resultados demonstram que o AG pode ser um novo agente terap?utico no controle da resposta exacerbada do corpo contra um agente infeccioso.

Made available in DSpace on 2015-04-14T13:35:51Z (GMT). No. of bitstreams: 1 453321.pdf: 2208938 bytes, checksum: 36179d9e8ff3571a84f104fd9b7bd8ef (MD5) Previous issue date: 2013-12-18 / Fibrosis is a chronic liver disease that is a major cause of human mortality and is characterized by the accumulation of extracellular matrix in response to chronic liver injury. Important causes of chronic liver injury are: viral hepatitis, metabolic diseases, autoimmune diseases and exposure to chemicals, such as alcohol or drugs. The GRX cells are a representative line of hepatic stellate cells (HSC), which is associated with development of fibrosis, in the last stage is the cirrhosis. In healthy liver, these cells exhibit a phenotype or quiescent lipocyte characterized by its hability to store lipid droplets. Gallic acid is involved in several biological processes such as cell growth inhibition and apoptosis also has a variety of pharmacological actions, including antioxidant activity, anti-inflammatory, antimicrobial and antitumor. The aim of this study was to investigate the in vitro effects of gallic acid on the phenotype of HSC. The results showed that gallic acid is able to reduce cell proliferation, induce quiescent phenotype in HSCs by increasing lipid droplets, probably by activating peroxisome proliferator-activated receptor gama, decrease of transforming growth factor 1 signaling and decreased expression of collagen type I. These results demonstrate that the gallic acid may be a novel therapeutic agent for treating hepatic fibrosis. / A fibrose ? uma doen?a cr?nica do f?gado que representa uma das maiores causas de mortalidade humana e ? caracterizada pelo ac?mulo de matriz extracelular em resposta ? les?o hep?tica cr?nica. Importantes causas de les?es hep?ticas cr?nicas s?o: hepatites virais, doen?as metab?licas, doen?as autoimunes e exposi??o a subst?ncias qu?micas, como ?lcool ou drogas. As c?lulas GRX s?o uma linhagem representativa das c?lulas estreladas hep?ticas (HSC), que est? associada ao desenvolvimento da fibrose que, em ?ltimo est?gio, ? a cirrose. No f?gado saud?vel, estas c?lulas apresentam um fen?tipo quiescente ou lipoc?tico, caracterizado pela sua capacidade de armazenar got?culas lip?dicas. O ?cido g?lico est? envolvido em v?rios processos biol?gicos, tais como a inibi??o do crescimento celular e apoptose, al?m de possuir uma variedade de a??es farmacol?gicas, incluindo as atividades antioxidantes, anti-inflamat?rias, antimicrobiana e antitumoral. O objetivo deste estudo foi investigar os efeitos in vitro do ?cido g?lico sobre o fen?tipo das HSC. Os resultados obtidos mostraram que o ?cido g?lico ? capaz reduzir a prolifera??o celular, induzir o fen?tipo quiescente nas HSCs pelo aumento de got?culas lip?dicas, provavelmente pela ativa??o do receptor ativado por proliferador de peroxissomo gama, bloqueio da sinaliza??o de fator de transforma??o do crescimento beta 1 e diminui??o da express?o do col?geno tipo I. Estes resultados demonstram que o ?cido g?lico pode ser um novo agente terap?utico para o tratamento de fibrose hep?tica.

Made available in DSpace on 2015-04-14T14:51:33Z (GMT). No. of bitstreams: 1 459268.pdf: 1089012 bytes, checksum: 3e3bbefbe5c53d842c73d0dec81136ac (MD5) Previous issue date: 2014-02-18 / Hepatocellular Carcinoma is the most prevalent primary tiver tumor and is among the top ten cancers that affect the world population. lts development is related, in most cases, the existente of chronic tiver injury, such as occurs in cirrhosis. Current curative therapies are only surgical resection, transplantation and percutaneous ablation, however with the possibility of recurrence. In this context, the search for new therapies for the disease becomes an interesting field for research. The knowledge about the correlation between chronic inflammation and cancer has driven new researches with anti-inflammatory agents that have potential for the development of antitumor drugs. Gallic acid is a polyphenol found in many natural products and have shown anti-inflammatory activity, anti-tumor, antimutagenic and strong antioxidant action. The purpose of this study was to investigate the effect of gallic acid on cell proliferation and inflammatory parameters of tiver carcinoma cells (HepG2), as well as to investigated the mechanisms involved. Cell viability was evaluated through MTT colorimetric assay and Trypan blue exclusion. Results showed that the gallic acid decreased proliferation of HepG2 celis in a dose and time dependent manner, without causing necrosis (LDH assay). We observed significant induction of apoptosis by PE Annexin V and 7-AAD assay and no interferente with the cell cycle using the FITC BrdU Flow Kit. The leveis of inflammatory mediators were measured by Cytometric Bead Array Human Inflammation Assay and observed a significant reduction in the leveis of interleukin-8 (pro-inflammatory and related to angiogenesis, invasiveness and metastasis) and increased leveis of interleukin-10 (anti-inflammatory and related to the induction of programmed cell death) and interleukin-12 (antiangiogenic and antimetastatic). We also evaluated the leveis of TGF by ELISA (Enzyme-Linked lmmunosorbent Assay) and no significant differences. According to these results, we believe that gallic acid has a strong potential as an anti-tumor agent. / O Carcinoma Hepatocelular ? o tumor hep?tico prim?rio mais prevalente e est? entre as dez principais neoplasias que afetam a popula??o mundial. O seu desenvolvimento est? relacionado, na maioria das vezes, a uma les?o hep?tica cr?nica, como ocorre na cirrose. As terapias consideradas curativas atualmente s?o somente a ressec??o cir?rgica, o transplante e a abla??o percut?nea, ainda com possibilidade de recidiva. Nesse contexto, a busca por novas alternativas terap?uticas para a doen?a torna-se um campo de pesquisa em expans?o. O conhecimento sobre a correla??o entre a inflama??o cr?nica e c?ncer tem impulsionado novas pesquisas com agentes anti-inflamat?rios que apresentem potencial para o desenvolvimento de drogas antitumorais. O ?cido g?lico ? um polifenol encontrado em v?rios produtos naturais, o qual tem apresentado atividade anti-inflamat?ria, antitumoral, antimutag?nica e forte a??o antioxidante. O objetivo desse estudo foi avaliar o efeito do ?cido g?lico sobre a prolifera??o celular e par?metros inflamat?rios das c?lulas de carcinoma hep?tico (HepG2), assim como investigar os mecanismos envolvidos. A viabilidade celular foi avaliada atrav?s do ensaio colorim?trico MTT e contagem celular por exclus?o com Azul de Trypan. Os resultados mostraram que o ?cido g?lico reduziu a prolifera??o celular de maneira dose e tempo dependente, sem causar necrose (ensaio LDH). Observamos indu??o significante da apoptose atrav?s do ensaio PE Annexin V and 7-AAD e nenhuma interfer?ncia no ciclo celular utilizando o kit FITC BrdU Flow. Os n?veis de mediadores inflamat?rios foram medidos utilizando o kit Cytometric Bead Array Human Inflammation. Observamos redu??o significante nos n?veis da interleucina 8 (pr?-inflamat?ria e relacionada ? angiog?nese, invasividade e met?stases), aumento dos n?veis de interleucina 10 (anti-inflamat?ria e relacionada ? morte celular programada) e aumento dos n?veis de interleucina 12 (antiangiog?nica e antimetast?tica). N?s tamb?m avaliamos os n?veis de TGF(3 por ELISA (Enzyme?Linked lmmunosorbent Assay) e n?o observamos diferen?as significativas. A partir desses resultados, acreditamos que o ?cido g?lico tem forte potencial como agente antitumoral.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-01-14T20:02:01Z No. of bitstreams: 1 MoacirFernandesDeQueirozNeto_DISSERT.pdf: 2451986 bytes, checksum: 86a4ed116bac73f635a0765a8cdd98c9 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-01-18T21:50:30Z (GMT) No. of bitstreams: 1 MoacirFernandesDeQueirozNeto_DISSERT.pdf: 2451986 bytes, checksum: 86a4ed116bac73f635a0765a8cdd98c9 (MD5) / Made available in DSpace on 2016-01-18T21:50:30Z (GMT). No. of bitstreams: 1 MoacirFernandesDeQueirozNeto_DISSERT.pdf: 2451986 bytes, checksum: 86a4ed116bac73f635a0765a8cdd98c9 (MD5) Previous issue date: 2014-12-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Quitina ? segundo polissacar?deo mais abundante na natureza e seu derivado quitosana tem sido amplamento estudado, devido a suas propriedades qu?micas e farmacol?gicas singulares. Contudo, estudos mostram que essa mol?cula quando no organismo, tende a se acumular no tecido renal e tamb?m promove um aumento na excre??o de c?lcio. Apesar disso, o efeito da quitosana sobre a forma??o de cristais de oxalato de c?lcio (OxCa) nunca foi avaliado. A forma??o de c?lculos renais (urolit?ase) ? a enfermidade que mais comumente afeta os rins e o sistema urin?rio, al?m de ser uma doen?a que possui altas preval?ncia e recorr?ncia. Muitas mol?culas com capacidade antioxidante tem demonstrado potencial para diminuir a forma??o de cristais de OxCa in vitro. Diante do exposto o objetivo desse trabalho foi avaliar o potencial antioxidante e de interfer?ncia na forma??o de cristais de uma quitosana de baixo peso molecular e seus derivados conjugados com ?cido g?lico (AG). As analises f?sico-qu?micas confirmaram a identidade da quitosana. Esta foi submetida a 5 testes antioxidantes e apresentou uma excelente atividade quelante de cobre, por?m nenhuma outra atividade antioxidante expressiva foi detectada. J? quando submetida aos testes de forma??o de cristais in vitro, a quitosana aumentou o n?mero de cristais de OxCa monohidratados formados, modificou a morfologia desses cristais, modificou as propor??es entre popula??es de cristais em solu??o e aumentou o potencial zeta desses cristais formados. Foram obtidas quatro mol?culas de quitosana conjugadas com AG. As an?lises f?sico-qu?micas confirmaram que houve liga??o covalente entre quitosana e AG, por?m, n?o se observou uma conjuga??o de AG de forma dose-dependente. Quando estes derivados foram submetidas a testes antioxidantes, todas as quitosanas conjugadas apresentaram potencial antioxidante maior que seus percussores. Contudo, houve diferen?a de atividade entre as diferentes quitosanas conjugadas, indicando que a posi??o onde o AG est? conjugado ? um fator importante para determina??o da atividade. Quando as quitosanas conjugadas foram submetidas aos testes de forma??o de cristais in vitro, houve uma redu??o na quantidade de cristais observados quando comparados com aqueles formados na presen?a da quitosana n?o-conjugada. A quitosana possui uma forte capacidade indutora de forma??o de cristais de OxCa monohidratados, bem como modifica sua morfologia e potencial zeta. O processo de conjuga??o de AG ? quitosana levou a um aumento no potencial antioxidante dessa mol?cula e tamb?m foi capaz de diminuir sua capacidade de induzir ? forma??o de cristais in vitro / Chitin is the second most abundant polysaccharide in nature and its derivative chitosan has been widely studied due to its unique chemical and pharmacological properties. However, studies show that when this molecule is used as food, drug, etc. it tends to accumulate in renal tissue and promotes an increase in calcium excretion. Nevertheless, the effect of chitosan on the formation of calcium oxalate (OxCa) crystals has never been evaluated. The formation of kidney stones (urolithiasis) is the disease that most often affects the kidneys and the urinary system. In addition, this is a disease with high prevalence and recurrence. Many molecules with antioxidant activity have been shown to decrease the potential for in vitro OxCa crystals formation. Thus, the aim of this study was to evaluate the effect of low molecular weight chitosan and its derivatives conjugated to gallic acid (AG) as antioxidant and inhibitor of OxCa crystals formation. The physico-chemical analysis confirmed the identity of chitosan. This molecule was subjected to five antioxidant tests and showed an excellent copper chelating activity. However, chitosan did not show other significant antioxidant activity. When chitosan was subjected to in vitro crystal formation tests, it increased the number of OxCa monohydrate crystals, modified the morphology of the crystals, modified the proportions between populations of crystals in solution and increased the zeta potential of these crystals formed. Four molecules of chitosan conjugated with GA were obtained. The physico-chemical analysis confirmed that chitosan and AG were covalently bonded. However, the amount of GA liked to chitosan did not increase even when 10 times more GA was used in experiment. When these derivatives were subjected to antioxidant tests, all chitosan conjugates showed higher antioxidant potential than their precursors. However, they showed different activity between them, which indicating that the position where AG is conjugated is an important factor for chitosan-GA activity. When conjugated chitosans were submitted to in vitro crystal formation tests, a reduction in the crystals number was observed when compared with those formed in the presence of unconjugated chitosan. Chitosan has a strong capacity for inducing OxCa monohydrate crystal formation, as well as modify their morphology and zeta potential. Over all, the process of conjugating AG to chitosan led to an increase in antioxidant potential of this molecule and was also able to decrease its capacity of inducing in vitro crystal formation

Made available in DSpace on 2014-12-17T14:16:30Z (GMT). No. of bitstreams: 1 BeatrizPB_DISSERT.pdf: 1734874 bytes, checksum: 6c9183abd4605927f73899354ef306e2 (MD5) Previous issue date: 2012-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The herbal medicine Sanativo? is produced by the Pernambucano Laboratory since 1888 with indications of healing and hemostasis. It is composed of a fluid extract about Piptadenia colubrina, Schinus terebinthifolius, Cereus peruvianus and Physalis angulata. Among the plants in their composition, S. terebinthifolius and P. colubrina have in common phenolic compounds which are assigned most of its pharmacological effects. The tannins, gallic acid and catechin were selected as markers for quality control. The aim of this study was the development and validation of analytical method by HPLC/UV/DAD for the separation and simultaneous quantification of gallic acid (GAC) and catechin (CTQ) in Sanativo?. The chromatographic system was to stationary phase, C-18 RP column, 4,6 x 150 mm (5 mm) under a temperature of 35 ? C, detection at 270 and 210 nm. The mobile phase consisted of 0.05% trifluoroacetic acid and methanol in the proportions 88:12 (v/v), a flow rate of 1 ml/min. The analytical method presented a retention factor of 0.30 and 1.36, tail factor of 1.8 and 1.63 for gallic acid and catechin, respectively, resolution of 18.2, and theoretical plates above 2000. The method validation parameters met the requirements of Resolution n ? 899 of May 29, 2003, ANVISA. The correlation coefficient of linear regression analysis for GAC and CTQ from the standard solution was 0.9958 and 0.9973 and when performed from the Sanativo? 0.9973 and 0.9936, the matrix does not interfere in the range 70 to 110 %. The limits of detection and quantification for GAC and CQT were 3.25 and 0.863, and 9.57 and 2.55 mg/mL, respectively. The markers, GAC and CQT, showed repetibility (coefficient of variation of 0.94 % and 2.36 %) and satisfactory recovery (100.02 ? 1.11 % and 101.32 ? 1.36 %). The method has been characterized selective and robust quantification of GAC and CTQ in the Sanativo? and was considered validated / O medicamento fitoter?pico Sanativo? ? produzido pelo Laborat?rio Pernambucano desde 1888 com indica??es de cicatrizante e hemost?tico. Trata-se de um extrato fluido composto por Piptadenia colubrina, Schinus terebinthifolius, Cereus peruvianus e Physalis angulata. Dentre as plantas de sua composi??o, S. terebinthifolius e P. colubrina possuem em comum compostos fen?licos aos quais ? atribu?da a maior parte de seus efeitos farmacol?gicos. Os taninos, ?cido g?lico e catequina foram selecionados como marcadores para controle de qualidade desse medicamento. O objetivo do presente trabalho foi o desenvolvimento e valida??o de m?todo anal?tico por CLAE/UV/DAD para separa??o e quantifica??o simult?nea de ?cido g?lico (ACG) e catequina (CQT) no fitoter?pico Sanativo?. O sistema cromatogr?fico teve como fase estacion?ria, coluna RP C-18, 4,6 x 150 mm (5 μm), sob a temperatura de 35 ?C, detec??o em 270 e 210 nm. A fase m?vel foi composta por ?cido trifluoroac?tico 0,05 % e metanol nas propor??es 88:12 (v/v), sob vaz?o de 1 ml/min. O m?todo anal?tico desenvolvido apresentou um fator de reten??o de 0,30 e 1,36, fator de cauda de 1,8 e 1,63, para ?cido g?lico e catequina, respectivamente, resolu??o de 18,2, al?m de pratos te?ricos acima de 2000. O m?todo atendeu os par?metros de valida??o exigidos na Resolu??o RE n? 899, de 29 de maio de 2003, da ANVISA. O coeficiente de correla??o da an?lise de regress?o linear para ACG e CQT a partir da solu??o padr?o foi de 0,9958 e 0,9973 e a partir do Sanativo? foi de 0,9973 e 0,9936, a matriz do fitoter?pico n?o interfere no intervalo de 70 a 110%. Os limites de detec??o e de quantifica??o para ACG e CTQ foram de 3,25 e 0,863, e 9,57 e 2,55 μg/mL, respectivamente. Os marcadores, ACG e CTQ, apresentaram repetibilidade (coeficientes de varia??o de 0,94 % e 2,36 %) e recupera??o satisfat?ria (100,02 ? 1,11 % e 101,32 ? 1,36 %). O m?todo ainda se caracterizou seletivo e robusto para quantifica??o de ACG e CQT no Sanativo? e foi considerado validado