Thesis (DTech (Biomedical Technology))--Cape Peninsula University of Technology, 2009 / Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), which leads to neuronal demyelination, and eventually to oligodendrocyte and axon loss, with subsequent lesion formation. The wide distribution of lesions in the CNS results in a variety of clinical features, such as cognitive impairment, vertigo, spasticity, ataxia tremors, progressive quadriparesis, pain and depression. Currently no cure exists for CNS disorders, resulting in a decline in quality of life, and an economic burden on society. Metabolic disturbances, especially lipid metabolic abnormalities, have been implicated in the development of MS. Although the disease cannot be cured, disease-modifiers, such as interferon beta, glatiramer acetate and mitoxantrone, as well as fatty acid supplementatlon have been used to delay the progression of the disease. Membrane fatty acids are precursors for mediators of inflammation, the eicosanoids, which are produced soon after stimulation and which regulate a number of inflammatory effects, such as the induction of fever, vasodilation and production of macrophage- and Iymphocyte-derived cytokines. Eicosanoids, in contrast to their fatty acid precursors, have a short half-life and are therefore difficult to measure. The objective in the present study was to determine the role of fatty acids from South African MS patients, by measuring the fatty acid composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI) and sphingomyelin (SM) phospholipids in the plasma, red blood cell (RBC) and peripheral blood mononuclear cell (PBMC) membranes and correlate abnormalities with the neurological outcome as measured by the Kurtzke Expanded Disability Status Scale (EDSS) and inflammation assessed by C-reactive protein (CRP). A second objective was to establish whether possible changes in membrane lipids (phospholipids, fatty acids and cholesterol) would have an effect on membrane fluidity, and whether this would correlate with the EDSS and CRP. The plasma, RBC and PBMC membrane lipid composition from 31 white female patients with MS and 30 age- and gender-matched control subjects were assessed. Fatty acids were quanflfied by gas chromatography (GC), phospholipids by colorimetric and cholesterol by enzymatic assays. Membrane fluidity, as measured by the membrane lipid composition, was calculated, using previously established formulae, and includes the following: the saturated nature of the membrane was measured by the phospholipid PC+PS/PE+PS ratio, fluidity and permeability were measured by the cholesterol concentratlon and the cholesterol to total phospholipid ratio and membrane deformability was measured by the phospholipid PE to PS ratio. Membrane fluidity was also measured by the ordered-erystalline-phase to liquidcrystalline- phase lipid composition, which correlates with the phospholipid PE to PC ratio. The membrane saturated (SATS) to polyunsaturated fatty acid (PUFA) ratio was further used as an indication- of the fluidity status of the membranes. CRP was measured in all participants using a Beckman nephelometer. In MS, the n-6 fatty acids, particularly C18:2n-6, C20:4n-6 and C22:4n-6, were significantly decreased in plasma, RBC and/or PBMC membranes. In addition, the relationship between C20:3n-6 and C20:4n-6 showed a metabolic disturbance in both RBC and PBMC membranes from patients with MS, as compared to the control group. C20:4n-6 showed significant inverse correlations with the EDSS and CRP in MS patients, indicating that loss of these fatty acids from membranes correlated with higher disability as well as with increased inflammation. There were significant increases in free fatty acids C18:2n-6 and C20:4n-6 in plasma from MS patients. Saturated fatty acids, SM C14:0 and PI C22:0 were significantly increased in PBMC membranes from MS patients, and SM C14:0, C16:0 and C20:0 showed inverse correlations with the Functional System Scores. In contrast, the longer-ehain SATS, C22:0 and C24:0 showed positive correlations with the Functional System Scores. Red blood cell membrane fluidity as measured by the SATS to PUFA ratio was significantly higher in patients than in controls. In patients with CRP ~ 5.00 Ilglml the ratio showed significant inverse correlation with disease outcome. The saturated nature correlated positively, whilst the .ordered-erystalline-phase to liquid-crystalline-phase lipid ratio correlated inversely with the Functional System Scores. In this study it was consistently shown that C20:4n-6, or its precursor and elongation products, C18:2n-6 and C22:4n-6 respectively, was lower in plasma, RBC and/or PBMC membranes from MS patients. Red blood cells lack the desaturase enzymes and depend on fatty acids sourced from the plasma. Therefore, lower C20:4n-6 in the RBC membranes from MS patients may be due to depleted plasma stores, or an indication of an increased demand of this fatty acid elsewhere. Furthermore, this study has demonstrated that lower RBC C20:4n-6, with an increase in plasma FFA C20:4n6, resulted in worse disease outcome, perhaps due to the pro-inflammatory effect of eicosanoid production. This. study also characterized the specific SATS, that is, longer-ehain SATS that may increase the risk of developing MS, as higher shorter-ehain SATS, C14:0 and C16:0 reflected better disease outcome, demonstrated by the inverse correlation with the EDSS and FSS. Lastly, this study has shown that in the presence of uncontrolled inflammation such as in MS, the altered lipid composition indirectly compromised cell membrane, structure and fluidity, and thereby contributed to the disease progression in MS patients.

The aim of the study was to determine the impact of diet (natural and formulated feed) on the chemical composition and fatty acids profile of the harvested fish. The content of dry matter in fish flesh resulting from the formulated diet was higher vs. the natural diet (23.94?1.24 % vs. 19.66?0.82 %) with nitrogenous compounds (60.24?2.82 % vs. 72.12?1.75 %), total fat content (24.81?4.51 % vs. 6.14?2.85 %) and ash (7.55?1.28 % vs. 10.54?1.53 %) respectively. The spectrum of fatty acids was determined by gas chromatography using Varian 3800 equipment. Tench fed on a formulated diet in the recirculating system had a significantly higher content (P< 0.05) of monounsaturated fatty acid (MUFA = 43.04?1.68 %) and n-6 polyunsaturated fatty acids (PUFA = 15.47?1.07 %) in their flesh compared to the flesh of fish reared in earth ponds on a natural diet - MUFA (32?5.29 %) and n-6 PUFA (13.6?1.66 %). Tench fed on a natural diet in earth ponds proved to have a significantly higher content (P< 0.05) of n-3 PUFA (16.8?4.38 %) and ? PUFA (30.3 ? 5.3 %) than tench reared in the recirculating system - PUFA n-3 (10.05?0.85 %) and ? PUFA (25.52?1.07%). The ratio n-3/n-6 for fish from earth ponds was 1.2; for fish from the recirculating system the ratio was 0.65. The results show a significantly higher composition of n-3 PUFA in flesh of tench from earth pond with natural food compared to fish on an intensive feeding diet in the recirculating system which showed a higher content of n-6 PUFA.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / Os Ãcidos graxos insaturados Ãmega-3 (&#969;-3) e Ãmega-9 (&#969;-9) possuem aÃÃo anti-inflamatÃria e antioxidante, enquanto os Ãmega-6 (&#969;-6) sÃo prÃ-inflamatÃrios. Este estudo verificou os efeitos do prÃ-condicionamento com misturas de Ãleos contendo baixa relaÃÃo &#969;-6/&#969;-3 e elevada relaÃÃo &#969;-9/&#969;-6, em modelo experimental de isquemia-reperfusÃo cerebral. Foram utilizados 42 ratos Wistar, divididos em dois grupos: Controle (n=24) e Teste (n=18). O grupo Controle foi subdividido em 4 grupos de 6 animais, cada: Simulado - Ãgua (Sim-Ãgua), Isquemia-ReperfusÃo - Ãgua (IR-Ãgua), Simulado - IsolipÃdico (Sim-IsolipÃdico) e Isquemia-ReperfusÃo - IsolipÃdico (IR-IsolipÃdico). Os animais receberam Ãgua ou uma mistura isolipÃdica com relaÃÃes &#969;-6/&#969;-3 = 8:1 e &#969;-9/&#969;-6 = 0,4:1 por via orogÃstrica, durante sete dias, conforme seus grupos. O grupo Teste foi subdividido em 3 grupos de 6 animais: IR-Mix1, IR-Mix2 e IR-Mix3. Os animais do grupo Teste receberam misturas oleosas com relaÃÃes &#969;-6/&#969;-3 = 1,4:1 e &#969;-9/&#969;-6 = 3,4:1 , diferindo apenas na fonte de &#969;-3: Mix1, contendo o Ãcido &#969;-3 &#945;-linolÃnico; Mix2, contendo os Ãcidos &#969;-3 &#945;-linolÃnico, eicosapentaenÃico e docosaexaenoico, e Mix 3, contendo os Ãcidos &#969;-3 &#945;-linolÃnico e docosaexaenÃico, administradas por via orogÃstrica, durante sete dias. No sÃtimo dia, os animais dos grupos IR-Ãgua, IR-IsolipÃdico, IR-Mix1, IR-Mix2 e IR-Mix3 foram submetidos à isquemia cerebral com oclusÃo bilateral das artÃrias carÃtidas comuns por 1 hora, seguida de reperfusÃo por 3 horas. Os animais dos grupos Sim-Ãgua e Sim-IsolipÃdico foram submetidos à operaÃÃo simulada. Ao final do experimento, todos os animais foram decapitados e seus cÃrebros fatiados para anÃlise histopatolÃgica da Ãrea CA3 do hipocampo. A morte neuronal foi quantificada pela contagem de neurÃnios vermelhos (NV). Constatou-se que a quantidade de NV no grupo IR-Ãgua (36,83  9,79) foi maior (P = 0,0046) que a observada do grupo Sim-Ãgua (17,67  8,48), bem como a quantidade de NV no grupo IR-IsolipÃdico (29,83  12,19) foi maior (P = 0,0459) que a observada no grupo Sim-IsolipÃdico (14,17  11,62). NÃo foi constatada diferenÃa na quantidade de NV entre os grupos Sim-Ãgua (17,67  8,48) e Sim-IsolipÃdico (14,17  11,62), ou entre os grupos IR-Ãgua (36,83  9,79) e IR-IsolipÃdico (29,83  12,19). A quantidade de NV no grupo IR-Mix1 (12,33  6,31) foi menor que a verificada nos grupos IR-Ãgua (36,83  9,79; P < 0,01) e IR-IsolipÃdico (29,83  12,19; P < 0,05). As quantidades de NV nos grupos IR-Mix2 (10,67  2,81) e IR-Mix3 (7,33  6,47) tambÃm foram menores que as verificadas nos grupos IR-Ãgua (36,83  9,79; P < 0,001) e IR-IsolipÃdico (29,83  12,19; P < 0,01). NÃo foram constatadas diferenÃas nas quantidades de NV entre os grupos IR-Mix1 (12,33  6,31), IR-Mix2 (10,67  2,81) e IR-Mix3 (7,33  6,47), entre si. Conclui-se que, independentemente da fonte de &#969;-3, o prÃ-condicionamento com misturas de Ãleos contendo baixa relaÃÃo &#969;-6/&#969;-3 e elevada relaÃÃo &#969;-9/&#969;-6, protege os neurÃnios contra as lesÃes de isquemia-reperfusÃo cerebral em modelo experimental. / Omega-3 (&#969;-3) and omega-9 (&#969;-9) unsaturated fatty acids are anti-inflammatory and antioxidant, while omega-6 (&#969;-6) fatty acids are pro-inflammatory. This study investigated the preconditioning effects of fatty acids mixes with low ratio &#969;-6/&#969;-3 and high ratio &#969;-9/&#969;-6, in a brain ischemia-reperfusion experimental model. Forty-two Wistar rats were aleatory assigned to two groups: Control (n=24) and Test (n=18). Control group was divided into 4 groups, each with 6 animals: Water-Simulated (Water-Sim), Water - Ischemia-Reperfusion (Water-IR), Isolipid-Simulated (Isolipid-Sim) and Isolipid - Ischemia-Reperfusion (Isolipid-IR). The animals received water or a isolipid mix with &#969;-6/&#969;-3 ratio of 8:1 and &#969;-9/&#969;-6 ratio of 0,4:1 by gavage, for 7 days, according to their groups. Test group was divided into 3 groups of 6 animals: Mix1-IR, Mix2-IR, and Mix3-IR. All animals from Test group received oil mixes with &#969;-6/&#969;-3 ratio of 1,4:1 and &#969;-9/&#969;-6 ratio of 3,4:1 , differing only on the &#969;-3 source: Mix1, with &#969;-3 linolenic acid; Mix2, with &#969;-3 linolenic, eicosapentaenoic and docosahexaenoic acids, and Mix 3, with &#969;-3 linolenic and docosahexaenoic acids, by gavage, for 7 days. At the 7th day, animals from Water-IR, Isolipid-IR, Mix1-IR, Mix2-IR, and Mix3-IR groups were subjected to 1-hour brain ischemia by occlusion of both common carotid arteries, followed by a 3-hour reperfusion. Animals from Water-Sim and Isolipid-Sim groups were submitted to a simulated operation. At the end of the experiment, all animals were decapitated and their brains were sliced and sent to histological analysis of the CA3 hippocampal region. Neuronal death was quantified by the red neurons (RN) count. It was found that the number of RN in Water-IR group (36.83  9.79) was higher (P = 0.0046) than the number observed in Water-Sim group (17.67  8.48), and similarly, the number of RN in Isolipid-IR group (29.83  12.19) was higher (P = 0.0459) than the number observed in Isolipid-Sim group (14.17  11.62). There was no difference between the amount of RN from Water-Sim (17.67  8.48) and Isolipid-Sim (14.17  11.62) groups, nor between Water-IR (36.83  9.79) and Isolipid-IR (29.83  12.19) groups. The number of RN in Mix1-IR group (12.33  6.31) was lower than the number seen in Water-IR (36.83  9.79; P < 0.01) and Isolipid-IR (29.83  12.19; P < 0.05) groups. The amounts of RN in Mix2-IR (10.67  2.81) and Mix3-IR (7.33  6.47) groups were also lower than the amounts observed in IR-Water (36.83  9.79; P < 0.001) and IR-Isolipid (29.83  12.19; P < 0.01) groups. There were no differences between the Mix1-IR (12.33  6.31), Mix2-IR (10.67  2.81) and Mix3-IR (7.33  6.47) groups. In conclusion, regardless of the source of &#969;-3, preconditioning with fatty acids mixes with low ratio &#969;-6/&#969;-3 and high ratio &#969;-9/&#969;-6, protects the neurons against brain ischemia-reperfusion injuries in this experimental model.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / The study was conducted to evaluate the influence of supplementation of different lipid sources on performance and quantitative characteristics of housing and not housing, fatty acid profile and muscle proteome Longissimus dorsi of Santa Ines lambs fed different additional sources of lipids. 35 lambs bulls were used, with initial body weight (13  1.80 kg) and approximately two months in a randomized block design with five treatments and seven repetitions. The treatments consisted of five diets, one free additional lipids (control) and the other, added cottonseed (CA), bran cashew (FCC), cottonseed with cashew nut meal (CALFCC) and calcium salts of long chain fatty acids (LCFA-Ca). The duration of the experiment was determined by the time required for the average body weight of all animals in each treatment were 28 kg, and this selected group for slaughter. No significant effect was observed with the use of supplementary sources of lipids on the loss of fasting, body weight at slaughter, empty body weight, hot carcass weight, cold carcass weight loss by cooling and biological yield. The non-carcass components, intestines and organs showed no differences across lipid additional sources. Regarding the fatty acid profile were observed 13 fatty acids, four of which are saturated, monounsaturated five-four polyunsaturated (PUFA). In proteome analysis, the proteins were expressed in greater quantity were located primarily at pH less than 5.5. Comparing the expression intensity spots between treatments can observe significant difference in the intensity of the spots 23 for the diet revealed fifteen proteins and of this total, seven were differentiated with respect to time. The proteins identified in this experiment can be mainly classified as structural cellular organization and protection to stress, as well as specific metabolic functions and other functions. The addition of various sources of lipid influenced the productive performance and features: hot carcass dressing and cold, yield and weight of the rack the weight and performance of the gastrointestinal tract filled, small intestine and liver income in Santa Ines sheep growing. Supplementation with different lipid sources influence the profile of fatty acids of Longissimus dorsi, so that diets containing FCC and Lcfa-Ca increased the amount of CLA in the muscle. The proteome analysis may be a method to identify marker proteins meat quality prediction. In this study, the identified proteins are highly relevant to the meat tenderness process, wherein the meat maturation process is complex and involves protein in different biological processes. The type of power had greater influence on the proportions of bodies responsible for digestion and absorption of nutrients. / O estudo foi realizado com objetivo de avaliar a influÃncia da suplementaÃÃo de diferentes fontes de lipÃdeos sobre o desempenho produtivo e as caracterÃsticas quantitativas de carcaÃa e nÃo carcaÃa, perfil de Ãcidos graxos e proteoma do mÃsculo Longissimus dorsi de cordeiros Santa InÃs alimentados com diferentes fontes suplementares de lipÃdeos. Foram utilizados 35 cordeiros nÃo-castrados, com peso corporal mÃdio inicial (13  1,80 kg) e aproximadamente, dois meses de idade em delineamento de blocos ao acaso com cinco tratamentos e sete repetiÃÃes. Os tratamentos experimentais consistiram de cinco raÃÃes, sendo uma isenta de lipÃdeos suplementares (controle) e as demais, adicionadas de caroÃo de algodÃo (CA), farelo da castanha de caju (FCC), caroÃo de algodÃo com farelo de castanha de caju (CALFCC) e sais de cÃlcio de Ãcidos graxos de cadeia longa (Ca-Agcl). A duraÃÃo do experimento foi determinada pelo tempo necessÃrio para que a mÃdia do peso corporal de todos os animais de cada tratamento atingisse 28 kg, sendo este grupo selecionado para o abate. NÃo foi observado efeito significativo com a utilizaÃÃo de fontes suplementares de lipÃdeos sobre a perda do jejum, peso corporal ao abate, peso de corpo vazio, peso de carcaÃa quente, peso de carcaÃa fria, perda por resfriamento e rendimento biolÃgico. Os componentes nÃo-carcaÃa, vÃsceras e ÃrgÃos, nÃo apresentaram diferenÃas em funÃÃo das fontes suplementares lipÃdicas. No tocante ao perfil de Ãcidos graxos foram observados 13 Ãcidos graxos, dos quais quatro sÃo saturados, cinco monoinsaturados e quatro poliinsaturados(AGPI). Na anÃlise do proteÃma as proteÃnas que foram expressas em maior quantidade se localizaram principalmente em pH inferior a 5,5. Comparando a expressÃo da intensidade de spots entre os tratamentos pode-se observar diferenÃa significativa na intensidade de 23 spots em funÃÃo da dieta revelando quinze proteÃnas sendo que desse total, sete foram diferenciadas em funÃÃo do tempo. As proteÃnas identificadas neste experimento podem ser classificadas principalmente como estruturais de organizaÃÃo celular e proteÃÃo ao stresse, bem como com especÃficas funÃÃes metabÃlicas e outras funÃÃes. A adiÃÃo de diferentes fontes de lipÃdeo suplementar influenciou o desempenho produtivo e as caracterÃsticas: rendimento da carcaÃa quente e fria, rendimento e peso da costela o peso e rendimento do trato gastrointestinal cheio, intestino delgado e rendimento do fÃgado em ovinos Santa InÃs em crescimento. A suplementaÃÃo com diferentes fontes lipÃdicas influenciou o perfil de Ãcidos graxos do Longissimus dorsi, de modo que as dietas contendo FCC e Ca-Agcl aumentou a quantidade de CLA neste mÃsculo. A anÃlise do proteoma pode ser um mÃtodo para identificar as proteÃnas marcadoras de prediÃÃo da qualidade da carne. Nesse estudo, as proteÃnas identificadas sÃo altamente relevantes para o processo de maciez da carne, sendo que o processo de maturaÃÃo da carne à complexo e envolve proteÃnas em diferentes processos biolÃgicos. O tipo de alimentaÃÃo teve maior influÃncia sobre as proporÃÃes dos ÃrgÃos responsÃveis pela digestÃo e absorÃÃo de nutrientes.

Orientador: Edgard Graner / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-17T08:53:18Z (GMT). No. of bitstreams: 1 Carvalho_MarcoAntonio_D.pdf: 4182553 bytes, checksum: 178a34f020339db79ce9dcd7b7ce717c (MD5) Previous issue date: 2010 / Resumo: O melanoma é, dentre as neoplasias malignas de pele, a de pior prognóstico devido ao seu alto potencial metastático e resistência aos agentes quimioterápicos existentes. A enzima metabólica ácido graxo sintase (FASN, EC2.3.1.85) é a responsável por catalisar a síntese de ácidos graxos saturados de cadeia longa. FASN é um homodímero com massa molecular de 250 kDa responsável pela produção do palmitato endógeno. Em tecidos normais, a atividade metabólica de FASN é mínima, uma vez que a maior parte dos ácidos graxos usados pelas células provém da dieta, com exceção dos tecidos lipogênicos. Por outro lado, tem sido demonstrado que nas células malignas a maior parte dos ácidos graxos provém da biossíntese endógena conseqüente ao aumento da atividade de FASN e que existe uma associação positiva entre a expressão desta enzima metabólica e o comportamento agressivo de tumores malignos, pois sua alta expressão ocorre principalmente em casos com prognóstico ruim. Inibidores específicos da atividade FASN bloqueiam a síntese de DNA e causam apoptose em linhagens celulares derivadas de neoplasias malignas de próstata, mama, cólon, estômago, intestino, endométrio, cavidade bucal, ovário e melanoma. A droga orlistat (Xenical®), aprovada pela FDA e utilizada para o tratamento de obesidade, foi descrita como tendo propriedades anti-neoplásicas em câncer de próstata, mama, cólon, estômago e melanoma, devido a sua capacidade de bloquear especificamente a atividade de FASN. Este trabalho teve como objetivo principal estudar o efeito do tratamento de camundongos C57BL6 com a droga orlistat sobre as atividades de MMPs, expressão de integrinas por células B16F10, adesão destas últimas à macromoléculas da MEC e formação de colônias pulmonares a partir da inoculação na veia caudal de camundongos C57BL6. Através de ensaios zimográficos, não observamos alterações das atividades de MMP-2 e -9 em células B16F10 tratadas com orlistat. No entanto, a inibição de FASN aumentou a adesão das células B16F10 aos componentes de matrix extracelular laminina e fibronectina. Através de ensaios de imunofluorescência observamos uma redução das regiões de adesão focal das integrinas ?v?3 nas células B16F10 tratadas com orlistat. Finalmente, o tratamento com orlistat reduziu em 53,6% o número de colônias metastáticas pulmonares, em comparação aos grupos controle. Em conjunto, os resultados aqui descritos sugerem que esta FASN é um alvo terapêutico em potencial para estes tumores. / Abstract: Malignant melanoma has poor prognosis due to its high metastatic potential and resistance to the existing chemotherapeutic agents. Fatty acid synthase (FASN, EC2.3.1.58) is a metabolic enzyme with molecular mass of 250 kDa responsible for the endogenous biosynthesis of saturated long chain fatty acids. FASN activity is relatively low in most normal human tissues, since most of the fatty acids used by the cells come from the diet, except in liver, adipose tissue, fetal lung and lacting breast. On the other hand, it has been demonstrated that in several cultured malignant cells fatty acids are mostly produced by FASN. A similar phenomenon is also observed in melanoma cells and overexpression of FASN has been associated with a poor prognosis for patients with this malignancy. Specific inhibitors of FASN activity block DNA synthesis and cause apoptosis in prostate, breast, colon, stomach, endometrial, oral cavity, ovary and melanoma cancer cells lines. Orlistat (Xenical®), approved for FDA and used for the treatment of obesity, has antitumor properties in prostate, breast, colon, gastric cancers and melanoma, due to its capacity to block the FASN activity. This work had as main objectives to study the effect of orlistat on the expression and activity of MMPs and expression of integrins in B16F10 cells, as well as on the adhesion of these cells to ECM macromolecules fibronectin and laminin. Moreover, we sought to verify the effect of this drug on the lung colonization by B16F10 innoculated in the tail vein of C57BL6 mice. The treatment with orlistat did not change MMP-2 and -9 gelatinolytic activities in B16F10 cells and enhanced the adhesion of these cells on laminin or fibronectin. Interestingly, treatment of B16F10 cells with orlistat promoted a reduction on the number of integrin ?v?3 focal adhesion plates observed in the immunofluorescence assay. Finally, orlistat promoted an inhibition of 53,6% in the number of lung metastatic foci, in comparison with the control groups, further confirming the anticancer potential of FASN inhibitors. Finally, the results here described suggest that FASN is a therapeutical target in potential for these tumors. / Doutorado / Patologia / Doutor em Estomatopatologia

Orientador: Edgard Graner / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-09T18:30:48Z (GMT). No. of bitstreams: 1 Carvalho_MarcoAntonio_M.pdf: 17374966 bytes, checksum: 8afc90916c0a8c3ff41261e0bd5b620d (MD5) Previous issue date: 2007 / Resumo: O melanoma é, dentre as neoplasias de pele, a de pior prognóstico devido ao seu alto índice de metástases. A ácido graxo sintase (FAS) é uma enzima metabólica formada por um homodímero com massa molecular aproximada de 250 kDa responsável pela síntese de ácidos graxos saturados de cadeia longa. Sua expressão é baixa em tecidos normais e alta em muitos tumores malignos, nos quais parece estar associada com um pior prognóstico para o paciente, como ocorre nos melanomas. Existem evidências que, nas células malignas, a atividade de FAS é necessária para a produção de fosfolipídios que compõem as membranas celulares. Inibidores específicos da atividade de FAS bloqueiam a síntese de DNA e causam apoptose em linhagens celulares derivadas de neoplasias malignas de próstata, mama e cólon. A droga Orlistat (Xenical®), usada para tratamento de obesidade, foi recentemente descrita como tendo propriedades anti-neoplásicas em câncer de próstata e mama, devido à sua capacidade de bloquear especificamente a atividade de FAS. Este trabalho teve como objetivo principal estudar o efeito da inibição da enzima FAS em células B16F10 e em um modelo experimental de metástases espontâneas de melanoma em camundongos. O tratamento de camundongos portadores de melanomas intraperitoneais causados pela injeção de células B16F10 com Orlistat reduziu em 50% o número de metástases espontâneas para os linfonodos mediastínicos. Experimentos de citometria de fluxo mostraram que o tratamento da linhagem celular B16F10 com Orlistat inibe a proliferação devido ao acúmulo de células em G0/G1 e bloqueio da passagem para a fase S, que foi confirmado por um aumento na quantidade da proteína p27Kip1 e concomitante redução de Skp2. Através de ensaios de marcação com Anexina-V, observamos que Orlistat provoca um aumento na taxa de apoptose destas mesmas células. O presente modelo animal de metástases de melanoma mostrou-se útil para o estudo de drogas com potencial efeito anti-tumoral, como o Orlistat. A inibição da atividade de FAS tem um importante papel na disseminação metastática de melanomas de camundongos, possivelmente por inibir a proliferação e provocar apoptose nas células B16F10 / Abstract: Malignant melanoma is a cancer with poor prognosis due to its high metastatic potential and resistance to the existing chemotherapeutic agents. Fatty acid synthase is a metabolic enzyme with molecular mass of 250 kDa responsible for the endogenous biosynthesis of fatty acids. FAS converts acetyl-CoA and malonyl-CoA in the long chain fatty acid palmitate. FAS activity is downregulated in most normal human tissues except liver, adipose tissue, fetal lung and lacting breast. On the other hand, FAS is overexpressed in several malignant neoplasms, including malignant melanoma. For some tumors, FAS overexpression has been associated with a poor prognosis. Recently, the new inhibitor of FAS activity Orlistat was hown to inhibit the proliferation of prostate and breast cancer cell lines and the growth of their xenograft tumors. In this work we studied the effects of FAS inhibition in the B16F10 melanoma cell line and in a animal model for melanoma metastasis. B16F10 murine melanoma cells were intraperitoneally injected in 40 mice and an inhibition of 50% in the number of mediastinal lymph node metastasis was observed in the Orlistat treated group. The inhibition of FAS was confirmed by ¿Oil Red O¿ staining in frozen tissue sections. We also found that Orlistat promoted apoptosis and inhibited the proliferation of B16F10 cells by blocking cell cycle progression, as demonstrated by flow cytometry experiments. Western blotting analysis of protein extracts obtained from Orlistat-treated cells revealed overexpression of p27Kip1 and downregulation of Skp2. In conclusion, the present animal model is useful for the study of drugs with antitumoral effect like Orlistat. The inhibition of FAS has an important role on the metastatic spread of melanoma in C57BL6 mice by blocking the proliferation and inducing apoptosis in B16F10 cells. In addition, the inhibition of FAS by Orlistat represents a new approach in order to develop drugs for melanoma chemotherapy / Mestrado / Patologia / Mestre em Estomatopatologia

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Este estudo teve como objetivo determinar a composiÃÃo centesimal, o perfil de Ãcidos graxos e o valor calÃrico de trÃs espÃcies de crustÃceos do Estado do CearÃ. As espÃcies estudadas sÃo as mais comercializadas: lagosta espinhosa (Panulirus argus), camarÃo branco (Litopenaeus vannamei), caranguejo uÃÃ, (Ucides cordatus).Os resultados obtidos demonstraram elevados teores de umidade na vÃscera de caranguejo. As carnes de lagosta e de caranguejo apresentaram os maiores percentuais de proteÃna. Os teores de lipÃdios totais foram maiores no hepatopÃncreas de lagosta.Quanto Ãs cinzas, os maiores valores foram detectados na cabeÃa de camarÃo e na carne de caranguejo. O maior valor calÃrico foi encontrado no hepatopÃncreas de lagosta. Na carne e hepatopÃncreas de lagosta destacaram-se os Ãcidos graxos: palmÃtico, esteÃrico, olÃico. Na fraÃÃo de lipÃdios neutros de cabeÃa de camarÃo, os quatros Ãcidos graxos majoritÃrios foram linolÃico, palmÃtico, olÃico e DHA. Nos fosfolipÃdios dessa amostra, os Ãcidos palmÃtico, linolÃico, olÃico e esteÃrico foram os majoritÃrios. / This study aimed to determine the chemical composition, the profile fatty acids and caloric value of three species of crustaceans of the State of CearÃ. The species are the most commercialized, spiny lobster (Panulirus argus), white shrimp (Litopenaeus vannamei), crab uÃÃ (Ucides cordatus). The results showed high levels of moisture in Crab viscera. The meat of lobster and crab presented higher percentages of protein. The levels of total lipids were higher in lagosta.Quanto hepatopancreas of the ash, the highest values &#8203;&#8203;were detected the head of shrimp and crab meat. The caloric value was greater found in the hepatopancreas of lobster. In meat and lobster hepatopancreas highlights are the fatty acids: palmitic, stearic, oleic. In the fraction of lipids neutral shrimp head, the four major fatty acids were linoleic, palmitic, oleic and DHA. Phospholipids in this sample, the fatty palmitic, linoleic, oleic and stearic acids were the major ones.

O objetivo deste estudo foi avaliar o efeito da competição de triathlon na função e morte de neutrófilos de atletas de elite e investigar o possível envolvimento dos ácidos graxos livres (AGs) neste processo. Os neutrófilos foram obtidos do sangue coletado de 11 sedentários e de 12 triatletas em repouso e após competição de triathlon (Half Ironman, 2 Km de natação, 80 Km de ciclismo e 20 Km de corrida). A competição de triathlon aumentou a capacidade dos neutrófilos de migrar e de realizar burst oxidativo, porém inibiu a fagociose realizada por estas células. Além disso, induziu aumento da fragmentação de DNA e externalização de fosfatidilserina. A competição de triathlon aumentou a concentração de AGs no plasma dos triatletas e esta foi correlacionada positivamente com a proporção de neutrófilos com DNA fragmentado e fosfatidilserina externalizada. A elevação da concentração plasmática dos ácidos oléico, linoléico e esteárico induzida pela competição parece estar envolvida nas alterações funcionais e na apoptose verificadas após a competição de triathlon. / The objective of this study was to evaluate the effect of triathlon competition on function and death of neutrophils from elite athletes and to investigate the involvement of fatty acids in this process. Neutrophils were obtained from blood collected from eleven sedentary volunteers and twelve triathletes under rest and after a Half Ironman triathlon competition (2 Km swimming, 80 Km cycling and 20 Km running). The triathlon competition increased the migration and reactive oxygen species production in neutrophils, however it reduced the phagocytosis activity. Moreover, it induced neutrophils death possibly by apoptosis as indicated by DNA fragmentation and phosphatidylserine externalization. The increase in plasma levels of oleic, linoleic and stearic acids induced by the competition may be involved in modulation on neutrophil function and death.

Introdução: Evidências experimentais, epidemiológicas e clínicas mostram o papel da inflamação na patogênese de desordens metabólicas, sendo a modulação da resposta inflamatória associada a quantidade e a qualidade dos ácidos graxos (AG) da dieta. Polimorfismos de nucleotídeo único (SNP) podem influenciar a relação entre AG e concentração plasmática de biomarcadores inflamatórios. Objetivo: Verificar a associação de SNP relacionados aos genes da adiponectina, Receptor do tipo Toll (TLR)-4, interleucina (IL)-1 e IL-6 e ingestão de lipídios com um padrão inflamatório sistêmico, baseado na concentração plasmática de onze biomarcadores inflamatórios em estudo de base populacional ISA-Capital. Metodologia: O presente estudo compreende adultos (20 a 59 anos) do estudo de base populacional, ISA-capital 2008-2010 (n=302). A coleta dos dados dietéticos foi realizada por meio de recordatório de 24 horas, aplicado em duplicata. A partir do plasma, foram determinadas as concentrações plasmáticas de adiponectina, proteína C reativa (PCR), IL-1, IL-6, IL-8, IL-10, fator de necrose tumoral-alfa, IL- 12p70, Quimiocina C-C motif ligante (CCL) 2, molécula de adesão intercelular solúvel (sICAM)-1 e molécula de adesão celular vascular solúvel (sVCAM)-1, por meio da técnica multiplex de imunoensaio, e o perfil de ácidos graxos (AG) do plasma por cromatografia gasosa. A partir do DNA genômico foi realizada a genotipagem dos SNP relacionados aos genes da adiponectina (rs266729, rs17300539, rs16861209, rs1501299 e rs2241766), TLR4 (rs4986790, rs4686791, rs5030728, rs11536889), IL-1 (rs1143623, rs16944, rs1143627, rs1143634 e rs1143643) e da IL-6 (rs1800795, rs1800796, rs1800797) pelo sistema Taqman Open Array. Uma análise multivariada de Cluster (k-means) foi realizada para separar os indivíduos legíveis entre grupo inflamado (INF), n=93, e grupo não inflamado (NINF),n=169, segundo a concentração plasmática dos onze 8 biomarcadores inflamatórios avaliados. Resultados: Todos os SNP estavam em equilíbrio de Hardy-Wienberg (n=301). O INF apresentou idade, circunferência da cintura, pressão arterial e concentrações de triacilgliceróis sanguíneo estatisticamente maiores que aqueles observados para o NINF. O INF apresentou concentração plasmática de AG palmítico (C16:0), razões AG saturados (AGS)/AG ômega-6 (n-6) e AGS/ AG poli-insaturados (AGPI) e atividade estimada da enzima estearoil CoA desaturase aumentadas e concentrações plasmática de AGPI, n-6 e AG araquidônico (AA) e atividade estimada da enzima delta-5-dessaturase (D5D) reduzidas em comparação com o NINF. Interações SNP-AG plasmáticos estatisticamente significantes para predisposição ao padrão inflamatório sistêmico foram detectadas entre o SNP +6054 G>A (rs1143643) do gene da IL-1 e os AG esteárico, AA e AGPI e atividade estimada da enzima delta-6-desaturase (D6D); entre o SNP +3725 G>C (rs11536889) do gene do TLR-4 e a razão AA/AG eicosapentaenoico (EPA); entre o SNP +45 T>G (rs2241766) do gene da adiponectina e o AG ômega-3 (n-3); entre o SNP -7734 C>A (rs16861209) do gene da adiponectina e AA; e entre o SNP -11391 G>A (rs17300539) do gene da adiponectina e AGS. Conclui-se que algumas frações dos AG do plasma podem modular a inflamação e que SNP localizados nos genes da adiponectina, TLR-4, IL- 1 e IL-6 podem interagir com as frações de AG do plasma influenciando a chance de desenvolver uma inflamação sistêmica. / Introduction: Experimental, epidemiological and clinical evidences point to a pathogenic role of inflammation on metabolic disorders development, and to the relationship between this inflammatory response and the quantity and quality of dietary fatty acids. Single Nucleotide Polymorphisms (SNP) can modulate the relationship between fatty acids and plasma inflammatory biomarkers levels. Objective: To verify the association between SNP in the genes of adiponectin, TLR- 4, IL-1 and IL-6 and dietary fatty acids and their effects to a systemic inflammatory pattern at a population-based study ISA-Capital. Methods: This study sample was composed by adults (20 to 59 years), participants of the population-based study ISAcapital 2008-2010 (n=302). Dietary data was collected using two 24 hours dietary recall. Plasma concentration of adiponectin, C reactive protein, IL-1, IL-6, IL-8, IL- 10, tumor necrosis factor-alfa, IL-12p70, Chemokine C-C motif ligand (CCL) 2, soluble intercellular adhesion molecule (sICAM)-1 and soluble vascular cell adhesion molecule (sVCAM)-1 was determined by multiplex immunoassay. Plasma FA profile was determined by gas chromatography. SNP from adiponectin (rs266729, rs17300539, rs16861209, rs1501299 e rs2241766), TLR4 (rs4986790, rs4686791, rs5030728, rs11536889), IL-1 (rs1143623, rs16944, rs1143627, rs1143634 e rs1143643) and IL-6 (rs1800795, rs1800796, rs1800797) gene were genotyped by Taqman Open Array system. A Cluster multivariate analysis (k-means) was conducted to separate individual into inflammatory group (INF), n=93, and noninflammatory group (NINF), n=169, according to eleven inflammatory biomarkers plasma levels. Results: All the SNP were in Hardy-Weinberg equilibrium (n=301). INF had statistically higher age, waist circumference, blood pressure and plasma tryglicerides concentration than NINF. INF presented statistically higher plasma palmitic acid (C16:0) levels, saturated fatty acid (SFA)/omega-6 fatty acid (n-6) ratio and SFA/polyunsaturated fatty acid (PUFA) ratio and estimated stearoil CoA 10 desaturase activity, and statistically lower plasma PUFA, n-6 and arachidonic acid e (AA) and estimate delta-5-desaturase (D5D) activity in comparison to NINF. Statistically significant SNP-plasma fatty acid interactions were found between SNP +6054 G>A (rs1143643) of IL-1 gene and stearic acid, AA and PUFA and estimate delta-6-desaturase (D6D) activity; between SNP +3725 G>C (rs11536889) of TLR-4 gene and AA/eicosapentaenoic acid (EPA) ratio; between SNP +45 T>G (rs2241766) of adiponectin gene and omega-3 fatty acid (n-3); between SNP -7734 C>A (rs16861209) of adiponectin gene and AA; and between SNP -11391 G>A (rs17300539) of adiponectin gene and SFA. In conclusion, some plasma fatty acid subfractions can modulate inflammation and SNP of adiponectin, TLR-4, IL-1 and IL-6 genes can interact with plasma fatty acids to modulate the chance to develop systemic inflammation.

Orientadores: Maria Regina Wolf Maciel, Cesar Benedito Batistella / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-04T03:08:23Z (GMT). No. of bitstreams: 1 Martins_PatriciaFazzio_M.pdf: 4266626 bytes, checksum: a048f065c12ba7c99e232ca5dd0eb892 (MD5) Previous issue date: 2005 / Resumo: Os destiladores desodorizados de óleos vegetais (DDOV) são constituídos por diversas substâncias, entre elas, os ácidos graxos livres, fitoesteróis, tocoferóis, ésteres de fitoesteróis, hidrocarbonetos, glicerídeos e produtos da decomposição destas moléculas. O teor de ácidos graxos livres (AGL) nos DDOV varia entre 24 e 85%. Devido ao seu alto teor, a separação dos AGL dos destilados desodorizados dos óleos vegetais é uma etapa importante para proceder à concentração de tocoferóis e fitoesteróis em alta pureza. Os tocoferóis são substâncias naturais valiosas usadas em alimentos, cosméticos e fármacos devido às suas propriedades vitamínicas e antioxidantes. Os fitoesteróis têm recebido grande atenção ultimamente, devido à sua atividade anticolerolêmica e potencialidade de aplicação em nutracêuticos. Neste trabalho, foram realizados ensaios para estudar a remoção dos AGL presentes nos destilados desodorizados de óleos vegetais provenientes do refino de óleo de girassol (DDOG) e de canola (DDOC), utilizando o processo de destilação molecular. Para isso, diversas condições operacionais de temperatura do evaporador e da vazão de alimentação foram utilizadas. O objetivo é avaliar o comportamento do sistema e determinar as melhores condições operacionais para realizar a eliminação de ácidos graxos, minimizando as perdas de tocoferóis durante o processo... Observação: O resumo, na íntegra, poderá ser visualizado no texto completo da tese digital / Abstract: Distilates of the vegetable oil deodorization (DVOD) are composed by free fatty acids, phytosterols, tocopherols, phytosterol esters, hydrocarbons, acylglycerols and products of decomposition of these molecules. The content of free fatty acids (FFA) in the deodorizer distillates varies between 24% and 85%. Due to its high content, free fatty acid separation from deodorizer distillate is an important step to concentrate tocopherols and phytosterols in high purity. Tocopherols are valuable natural substances used in food, cosmetic and pharmaceutical industries due to its vitaminic and antioxidant properties. Phytosterols have received grat attention because its hipocholesterolemic properties and potential to be used in nutraceutics. In this work, separation of free fatty acids from sunflower (SuODD) and canola oil (CODD) deodorizer distillates was investigated through molecular distillation. For this, different operating conditions of evaporator temperature and feed flow rate were used. The intention is to evaluate the behavior of the system and to determine the best operating conditions to eliminate FFA, minimizing tocopherol losses during the process. Furthermore, in this work, it was necessary to establish analytical techniques to determine the contents of tocopherols, phytosterols, triacylglycerols, diacylglycerols, monoacylglycerols and free fatty acids in DVOD. Besides, of DDOC and DDOG, the soybean oil deodorizer distillate (SODD) was also characterized... Note: The complete abstract is available with the full electronic digital thesis or dissertations / Mestrado / Desenvolvimento de Processos Químicos / Mestre em Engenharia Química