Made available in DSpace on 2014-06-11T19:29:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-03-20Bitstream added on 2014-06-13T19:38:28Z : No. of bitstreams: 1 guardieiro_mm_me_botfmvz.pdf: 504146 bytes, checksum: 07979740dad67abc78d3a31a2e3dde43 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo do presente trabalho foi avaliar a resposta superovulatória, a produção embrionária e a resistência à criopreservação através de congelação ou vitrificação de embriões de novilhas Nelore suplementadas com gordura protegida ruminal (Megalac-E®). Foram utilizadas 40 novilhas da raça Nelore mantidas a pasto, divididas em dois grupos experimentais de acordo com o tipo de suplementação fornecido (G = concentrado com gordura protegida ruminal e C = suplementação controle, sem adição de gordura). Os suplementos foram formulados para serem isoenergéticos e isoprotéicos. O delineamento foi do tipo cross-over com 67 (primeira repetição) e 70 d (segunda repetição) de duração. Após 50 d de suplementação dietética, foi realizada a sincronização da emergência da onda para iniciar o protocolo de superovulação. Os embriões colhidos foram congelados ou vitrificados e avaliação de desenvolvimento embrionário in vitro foi realizada posteriormente. Não houve diferença (P > 0,10) entre os Grupos G e C em relação à resposta superestimulatória, ao número total de estruturas, embriões congeláveis, degenerados e ovócitos não fecundados colhidos. Apesar do grupo C ter mostrado maior reposta superovulatória que o G (15,7 ± 1,2 vs 18,0 ± 1,3 CL; P = 0,06), os embriões do Grupo C apresentaram maior taxa de eclosão, independentemente do método de criopreservação, às 48 h (33,1 ± 4,0%; n = 148 vs 17,3 ± 3,3%; n = 137; P = 0,009) e às 72 h (44,3 ± 4,2%; n = 148 vs 30,9 ± 4,0%; n = 137; P = 0,04) de cultivo do que os do grupo G. Além disso, os embriões vitrificados e congelados apresentaram taxas similares de eclosão (P > 0,10). Nas condições do presente estudo, o suplemento composto com gordura protegida não afetou a resposta superestimulatória e a produção de embriões, e os embriões do Grupo Controle foram mais tolerantes à criopreservação. / The aim of the current study was to evaluate the superovulatory response and embryo production, as well as embryo resistance to cryopreservation through freezing or vitrification of embryos from Nellore heifers supplemented with rumen-protected fat, Megalac-E®. Forty heifers kept in pasture were randomly divided into two experimental groups according to supplement source (G = supplement with rumen-protected fat and C = Control, without fat supplementation). The supplements were formulated to be isocaloric and isoproteic. Each female underwent both treatments in a cross-over design with approximately 67 d (first replicate) and 70 d (second replicate). After 50 d of feeding, the emergence of the wave was synchronized with the aid of hormones to iniciate the superovulation protocol. The embryos recovered were frozen or vitrified and subsequently in vitro embryo development evaluation was accomplished. There was no difference between G and C groups (P > 0.10) regarding superstimulatory response, number of total embryos/ova, viable embryos, degenerate embryos, or unfertilized oocytes recovered. However, group C had a greater superovulatory response than G (15.7 ± 1.2 vs 18.0 ± 1.3 CL; P = 0.06). Group C embryos presented greater hatching rate, independently of the cryopreservation method, at 48 h (33.1 ± 4.0%; n = 148 vs 17.3 ± 3.3%; n = 137; P = 0.009) and at 72 h (44.3 ± 4.2%; n = 148 vs 30.9 ± 4.0%; n = 137; P = 0.04) of in vitro culture. Moreover, vitrified and frozen embryos had similar hatching rate (P > 0.10). Under the conditions of the present study, supplementation with protected fat did not affect superstimulatory response, quantity or quality of embryos. However embryos from the Control group were more tolerant to cryopreservation.

No description available.

Dissertação composta por 3 artigos. / Este estudo foi conduzido na Universidade Tecnológica Federal do Paraná, Câmpus Dois Vizinhos, com o objetivo de avaliar dois sistemas de terminação e diferentes pesos de abate através do desempenho, componentes do corpo vazio e perfil de ácidos graxos presentes na carne de bezerros Holandês para produção de carne. Utilizaram-se 43 animais machos, não castrados da raça Holandês, com peso inicial de 57 kg e com 58 dias de idade. Destes animais, 23 foram confinados recebendo uma dieta constituída de silagem de milho, como volumoso e concentrado comercial à base de farelo de milho e farelo de soja, na relação 40:60. Os demais, 20 animais, foram terminados em pastagem cultivada, variando conforme o período do ano: azevém (Lolium multiflorum) no primeiro período, e nos dois subsequentes milheto (Pennisetum americanum) com suplementação do mesmo concentrado do confinamento a 1% do peso corporal. Em cada sistema, os animais foram distribuídos ao acaso em quatro diferentes pesos, pré-estipulados: 140; 180; 220 e 260 kg de peso corporal. A terminação em confinamento possibilitou redução da idade de abate em comparação à pastagem, que também atendeu às exigências nutricionais. Para avaliação do consumo a pasto, foi utilizado como marcador externo o óxido de cromo. O cálculo do consumo de nutrientes foi baseado na MS da dieta, logo o consumo de matéria seca foi de (5,65 contra 4,33 kg e 4,36 contra 3,32% do peso vivo), de NDT (3,70 contra 2,58 kg e 2,88 e 2,01% do peso vivo) e de FDA (0,98 contra 0,66 kg e 0,76 contra 0,51% do peso vivo), além do ganho médio diário (0,949 e 0,694 kg-1), todas as variáveis superiores (P<0,05) para os animais em confinamento em relação aos da pastagem. O consumo de PB, FDN e CA não apresentaram diferença significativa. Os bezerros holandeses possuem potencial para a produção de carne, expressando o melhor desempenho quando terminados em confinamento. O PCVZ apresentou crescimento linear com o aumento do peso de abate, assim como o rendimento de carcaça quente e fria e não houve efeito na relação PCVZ/PAB. O crescimento dos componentes externos da carcaça foi proporcional ao aumento de peso, cresceram aceleradamente nas fases iniciais e acompanharam o aumento de peso. Em unidades relativas apenas a cabeça e as patas. O peso absoluto do coração, rins, pulmão, fígado, baço, vesícula biliar, traqueia, aparelho reprodutivo, TGI e o conteúdo do TGI elevaram-se (P<0,05) linearmente. Em unidades relativas, o comportamento individual de alguns órgãos internos inverteu-se, com decréscimos frente ao aumento de peso, apenas o fígado, esôfago, TGI e o conjunto de componentes internos cresceu linearmente. O PCVZ não foi influenciado pela alimentação, as aparas e couro apresentaram significância (P<0,05) em pesos absolutos e o couro e a cabeça em pesos relativos. As diferenças apresentadas pelo fígado, traqueia, baço e conteúdo do TGI, foram suficientes para alterar os pesos absolutos e relativos dos componentes internos (P<0,05) observados nos animais a pasto, devido principalmente à alta proteína presente nas pastagens. Para análise dos ácidos graxos, a dieta utilizada nos sistemas de terminação não alterou os ácidos graxos saturados. Os ácidos insaturados C16:1n7 (Ácido palmitoleico) apresentaram maior teor em confinamento 1,02 contra 0,75g/g de lipídeos na pastagem, assim como o teor do Ácido y-linolenico (C18:3n6) mais elevado em confinamento 0,07 contra 0,04 g/g de lipídeos na pastagem e o e Ácido 5,8, II, 14,17-eicosapentaenoico (EPA) (C20:5n3), que variou de forma contrária com valores mais elevados na pastagem 0,45 contra 0,17 g/g de lipídeos em confinamento. Na totalidade o conjunto de ácidos graxos saturados, insaturados, poliinsaturados não sofreram variações quanto ao sistema de terminação utilizado, assim com o ômega 6 isoladamente. Apenas o ômega 3 possuem os maiores teores de alfa-linolênico (ω3). Desta forma reduziu a relação ω‑6/ω‑3, demonstrando a maior participação do ω-3 nas pastagens e redução do ω‑6 na carne de animais em pastagens. Para diferentes pesos de abates, os AGS e AGI decresceram frente ao aumento de peso dos animais. Aos 260 kg que apresentou maior participação do CLA e a melhor relação ω-6/ ω-3. Caracterizando a maior qualidade de carne de animais criados em pastagem e o melhor peso de abate é a partir dos 194 kg para a produção de vitelos. / This study was conducted at the Federal Technological University of Paraná, Campus Two neighbors, in order to evaluate two different finishing systems and slaughter weights through the performance, empty body components and fatty acids in the meat of calves to Dutch production meat. We used 43 male animals, not castrated Holstein breed, with initial weight of 57 kg and 58 days old. Of these animals, 23 were confined getting a diet consisting of corn silage as forage and commercial concentrate with corn meal and soybean meal, in relation 40:60. The remaining 20 animals, were completed in cultivated pasture, varying according to the time of year: ryegrass (Lolium multiflorum) in the first period, and the two subsequent millet (Pennisetum americanum) supplementation with the same concentrated confinement to 1% of body weight. In each system, the animals were randomly assigned to four different weights, pre-stipulated: 140; 180; 220 and 260 kg body weight. The feedlot finished possible reduction of slaughter age compared to pasture, which also met the nutritional requirements. For evaluation of the pasture consumption was used as an external marker chromium oxide. The calculation of nutrient intake was based on the diet DM, so the dry matter intake was (5.65 against 4.33 kg and 4.36 against 3.32% of body weight), NDT (3.70 against 2.58 kg and 2.88 and 2.01% of body weight) and FDA (0.98 against 0.66 kg and 0.76 against 0.51% of body weight), and the average daily gain (0.949 and 0.694 kg-1), all variables higher (P <0.05) in animals in feedlot in relation to the pasture. The consumption of CP, NDF and CA showed no significant difference. The Holstein calves have potential for the production of meat, expressing the best performance when finished in feedlot. The EBW showed linear growth with increasing slaughter weight, and the yield of hot and cold carcass was not affected in relation EBW / PAB. The growth of external components of substrate was proportional to the increased weight, grew rapidly in the early stages and accompanying weight gain. In relative units just head and feet. The absolute weight of the heart, kidney, lung, liver, spleen, gall bladder, trachea, reproductive tract, TGI and content amounted TGI (P <0.05) linearly. In relative units, individual behavior of some internal organs reversed, with decreases due to increased weight, only the liver, esophagus, TGI and the set of internal components increased linearly. The EBW was not influenced by food, chips and leather were significant (P <0.05) in absolute weights and the leather and the head relative weights. The differences presented by the liver, trachea, spleen and contents of TGI, were sufficient to alter the absolute and relative weights of the internal components (P <0.05) observed in animals on pasture, mainly due to the high protein present in pastures. For the analysis of fatty acids, the diet used in the termination systems did not change the saturated fatty acids. The C16 unsaturated acids: 1n7 (palmitoleic acid) had higher content of confinement 1.02 against 0.75 g / g lipid in the pasture, and on the content of linolenic acid-y (C18: 3n6) higher feedlot 0.07 against 0.04 g / g lipid in grassland eoe acid 5.8, II, 14,17-eicosapentaenoic (EPA) (C20: 5n3), which varied in a manner contrary with higher values in the pasture 0.45 against 0, 17 g / g lipid in confinement. In total the set of saturated fatty acids, unsaturated, polyunsaturated did not vary as to the termination system used and with the omega 6 alone. Only omega 3 have the highest levels of alpha-linolenic (ω3). Thus reduced the ω-6 ratio / ω-3, demonstrating the increased participation of ω-3 in the pastures and reducing the ω-6 in the flesh of animals in pastures. For different weights of slaughter, the SFA and UFA decreased due to increased weight of the animals. To 260 kg greatest participation of CLA and the best ω-6 ratio / ω-3. Featuring the highest quality of meat from animals raised on pasture and the best slaughter weight is from 194 kg for the production of calves. / 5000

Thesis (MTech (BioMedical Technology))--Cape Peninsula University of Technology, 2006 / In multiple sclerosis (MS) the myelin sheaths surrounding the axons in the brain are mainly affected by the disease process. Myelin consists for the most part of lipids and proteins. An abnormality in essential fatty acid metabolism is known to be present in patients with MS (Horrobin, 1979), reflected in a high ratio of n-6 to n-3 fatty acids in cell membranes. It has also been established previously that the pathogenesis of inflammatory disorders is aggravated by excessive consumption of n-6 fatty acids relative to n-3 fatty acids (Guesnet et al., 2005),and it has been shown that ingesting a larger proportion of n-3 fatty acids could be crucial in the regulation of cellular physiology and in the prevention of pathologies such as autoimmune and inflammatory diseases. Modern Western medical treatment for autoimmune diseases, which includes MS, involves the administration of immunosuppressive drugs, such as beta interferon, cortisone (prednisone), methotrexate and cytoxan, which reduce the effectiveness of the entire Immune system, and can have serious, sometimes life threatening, side effec1s (Perlmutter, 2006, htlp:/Iwww.msfac1s.org). It would therefore be of interest to investigate other options for treatment Although there is an extensive literature on fatly acids in MS, the actual details of the mechanisms of fatly add imbalances in MS have not been established. It would therefore be advisable to Investigate the abnormality of the MS cell membrane fatly acid profile. Previous studies focused on individual fatty acids, but it would be more relevant to investigate the relationships within and between the n-6 and n-3 series, and their effect on outcome, and to establish any possible cumulative effects, because the metabolism of fatty adds within the two series does have an effect on one another.

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