A região amazônica apresenta grande quantidade de plantas perenes, com particular relevância para as espécies frutíferas. Os frutos amazônicos são conhecidos pelo seu grande potencial energético e são usados como fonte de alimento nas grandes regiões do país. Dentre esses frutos, destaca-se o açaí, fruto do açaizeiro (Euterpe oleracea Martius), a copaíba (Copaifera officinalis L.) e a castanha do Pará (Bertholletia excelsa). Esses frutos são importantes para o desenvolvimento agroindustrial da região amazônica. Os mesmos contêm na sua composição proteínas, fibras e ácidos graxos. Os frutos possuem ácidos graxos são usados na área farmacêutica, com finalidade clínica e dermatológica. O projeto tem como objetivo apresentar técnicas analíticas de caracterização e quantificação dos ácidos graxos presente na composição oleosa dos frutos e descrever a ação farmacológica. Esses ácidos são o oléico, linoleico e palmítico. As técnicas analíticas com características de quantificação deverão gerar informações confiáveis e interpretáveis sobre a amostra, sendo o critério de avaliação denominado validação. A validação foi estudada neste trabalho, visando ter confiabilidade e reprodutibilidade nos resultados. Os processos analíticos com características de identificação e quantificação aplicados nesse projeto são: a cromatografia a gás acoplada a espectrometria de massas (CG/MS), a espectrometria de massas Tanden (MS/MS) e cromatografia líquida de alta eficiência com detector ultra violeta (HPLC/UV). Os resultados demonstram que os três frutos amazônicos comtem ácidos oléico, linoleico e palmítico em proporções diferentes potencializando seu uso em aplicações farmacêuticas, específicamente em tratamentos dermatológicos. / The Amazon region presents a great amount of perennial plants, with particular relevance to the fruit species. The Amazonian fruits are known for their high energy potential and are used as a food source in large regions of the country. Among these fruits, there is acai, the fruit of the açai palm (Euterpe oleracea Martius), copaiba (Copaifera officinalis L.) and Brazil nut (Bertholletia excelsa). These fruits are important for agro-industrial development in the Amazon region. They contain in their composition of protein, fiber and fatty acids. The fruits are fatty acids are used in the pharmaceutical field, with clinical and dermatological purposes. The project aims to provide analytical techniques for characterization and quantification of fatty acids present in the composition of the oily fruits and describe the pharmacological action. These acids are oleic, linoleic and palmitic acids. Analytical techniques for quantifying characteristics should generate reliable and interpretable information about the sample, and the evaluation criteria called validation. The validation was studied in this work, aiming to have reliability and reproducibility of the results. The analytical procedures to identify and quantify features implemented in this project are: a gas chromatography-mass spectrometry (GC / MS), Tanden mass spectrometry (MS / MS) and high performance liquid chromatography with UV detector (HPLC / UV). The results show that the three Amazon fruits did contain oleic, linoleic and palmitic acids at different increasing its use in pharmaceutical applications, specifically in dermatological treatments.

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Orientador: Silvia Helena Venturoli Perri / Coorientador: Gisele Zoccal Mingoti / Banca: Felipe Perecin / Banca: Flávia Lombardi Lopes / Banca: Caliê Castilho / Banca: Guilherme de Paula Nogueira / Resumo: O objetivo deste estudo foi avaliar os efeitos da suplementação do meio de cultivo com ácido linoleico e/ou catalase, em meio sem a adição de SFB, cultivados em atmosfera de baixa tensão de oxigênio sobre o desenvolvimento, qualidade e criotolerância de embriões bovinos. Em um grupo foi feita a suplementação com 100 μM de ácido linoleico (LA); 100UI de catalase (CAT) em um segundo, no terceiro grupo foram feitas as 2 suplementações (CAT+LA) e um quarto grupo sem suplementação (CONTROLE), durante todo o período de cultivo em atmosfera de 5%CO2, 7%O2 e 88%N2 em BAGS. As taxas de clivagem não diferiram (P>0,05) entre os grupos. Porém as taxas de produção de embriões em D7 e D8 diferiram (P<0,05), sendo que o controle obteve melhor resultado perante os 3 tratamentos e o grupo CAT+LA gerou a menor taxa de produção (30,7; 22,4; 19,8; 12,4%). Observou-se um atraso no desenvolvimento, sendo os blastocistos expandidos somente encontrados no D8. Na avaliação do conteúdo total de lipídios, houve uma redução significativa (P<0,05) nos 3 grupos tratados em relação ao controle. A medida dos níveis intracelulares de ROS não foi afetada nos 4 grupos (P>0,05). A quantidade de células totais foi significativamente menor (P<0,05) no grupo CAT+LA, e o grupo CAT foi o que apresentou maior porcentagem de células apoptóticas. Em relação à taxa de re-expansão às 24 horas (P<0,05), o grupo controle (50%) e o CAT (67,2%) apresentaram menores taxas que os grupos LA (71,7%) e o CAT+LA (76,7%). Às 48 horas, o grupo controle apresentou a menor taxa (35,7%), seguido pelo grupo CAT (47,5%). Já o grupo LA (56,5%) diferiu estatisticamente do controle mas não dos grupos CAT (47,5%) e CAT+LA (76,7%). O grupo CAT+LA diferiu estatisticamente dos grupos controle e CAT. Com base nestes resultados, embriões tratados com LA ou CAT+LA no cultivo in vitro, na ausêncio... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The most commonly recommended way to obtain more resistant in vitro embryos during the cryopreservation process is to change these embryos, whether through removal of fetal bovine serum (FBS), addition of different supplements to the culture medium or changes in oxygen tension in the production process. The objective of this study was to assess the effects of supplementation of the culture medium with linoleic acid and/or catalase in medium without the addition of FBS, in low tension oxygen atmosphere on development, quality and cryotolerance of bovine embryos. In one group, the medium was supplemented with 100 μm of linoleic acid (LA); the second was supplemented with 100UI of catalase (CAT), a third group was supplemented with both, CAT+LA and a fourth group received no supplementation (control), during the entire culture period in an atmosphere of 5% CO2, 7% O2 and N2 88% in BAGS. Cleavage rates did not differ (P>0.05) between groups (72.7; 74.1; 72.1; and 72.0%, respectively). However, embryo production rates in D7 and D8 were different (P<0.05), where the control had the highest blastocyst rate of the treatments and the CAT+LA group presented the lowest rate of production (30.7; 22.4; 19.8; 12.4%). Expanded blastocysts were only foundon D8, indicating a delay in development. There was a significant reduction (P<0.05) of total lipids in the 3 treated groups when compared to the control group. There was no difference in the intracellular levels of ROS between the 4 groups (P>0.05).Total cell number was significantly lower (P<0.05) in the CAT+ LA and the CAT group showed the highest percentage of apoptotic cells. Considering the re-expansion rates in 24 hours (P<0.05), the control group (50%) and the CAT group (67.2%) presented lower rates than the LA group (71.7%) and CAT+LA group (76.7%). At 48 hours, the control group showed the worst rate (35.7%) followed by the CAT group (47.5%)...(Complete abstract eletronic access below) / Doutor

Linoleic acid, with a DRI of 12-17g/d, is the most highly consumed polyunsaturated fatty acid in the Western diet and is found in virtually all commonly consumed foods. The concern with dietary linoleic acid, being the metabolic precursor of arachidonic acid, is its consumption may enrich tissues with arachidonic acid and contribute to chronic and overproduction of bioactive eicosanoids. However, no systematic review of human trials regarding linoleic acid consumption and subsequent changes in tissue levels of arachidonic acid has been undertaken. In this study, we reviewed the human literature that reported changes in dietary linoleic acid and its subsequent impact on changing tissue arachidonic acid in erythrocytes and plasma/serum phospholipids. We identified, reviewed, and evaluated all peer-reviewed published literature presenting data outlining changes in dietary linoleic acid in adult human clinical trials that reported changes in phospholipid fatty acid composition (specifically arachidonic acid) in plasma/serum and erythrocytes within the parameters of our inclusion/exclusion criteria. Decreasing dietary linoleic acid up to 90% was not significantly correlated with changes in tissue arachidonic acid levels (p=0.39). Similarly, when dietary linoleic acid levels were increased six fold, no significant correlations with tissue arachidonic acid levels were observed (p=0.72). However, there was a positive relationship between dietary gamma-linolenic acid and arachidonic acid on changes in tissue arachidonic levels. Our results do not support the concept that modifying current intakes of dietary linoleic acid has an effect on changing tissue levels of arachidonic acid in adults consuming Western-type diets.

The purpose of this thesis was to determine the combined effects of protein, creatine, and conjugated linoleic acid (CLA) supplementation during resistance training. These nutritional supplements are popular during resistance training and we wanted to determine if they would have additive effects for improving body composition and strength. Forty-four participants (32 males, 12 females, mean age 20y) were randomized into three groups to receive: 1) 36 g/d protein (PRO), 2) protein and 9 g/d creatine (PRO/CR), or 3) protein, creatine and 6 g/d CLA (PRO/CR/CLA) for 5 weeks while resistance training on a four-day cycle (three days of resistance training, followed by one day of rest). Measurements at pre- and post-testing included body composition, muscle thickness of the elbow and knee flexors and extensors, and bench and leg press strength. There were time main effects (p<0.01) for strength, and muscle thickness. The PRO/CR/CLA group had significant increases in knee extensor muscle thickness over time compared to the other groups (p<0.05). There were no other differences between groups over time. The combinations of creatine and protein, or creatine, protein and CLA had no effects on body composition. It is concluded that combining protein, creatine, and CLA has minimal effects on muscular strength, muscle thickness, and body composition.

El objetivo de esta tesis es: (1) determinar si el consumidor español asocia la grasa intramuscular (GIM) a la aceptabilidad de la carne de cerdo; (2) incrementar la GIM a través de estrategias nutricionales (adición de acido linoleico conjugado, reducción de vitamina A, reducción de proteína, lisina, suplementación con arginina y leucina). Los resultados mostraron que desde el punto de vista gustativo el consumidor prefiere la carne con un mayor contenido de GIM. Se obtuvo un incremento de la GIM al reducir el nivel de proteína (sin modificar la lisina) o al reducir el nivel de lisina (sin modificar la proteína) y al utilizar una línea genética grasa y no en una línea genética más magra. Se puede concluir que la modificación de la GIM a través de la dieta depende del genotipo, y que las modificaciones de los niveles de proteína y lisina son las más eficaces. / The objective of this thesis was: (1) to test if Spanish consumers associate intramuscular fat (IMF) content with acceptability of pork meat; (2) to increase IMF through nutritional strategies (supplementation with conjugated linoleic acid, reduction of vitamin A, reduction of protein, lysine, supplementation with arginine and leucine). Results showed that from the point of view of taste, consumers prefer the meat with a high IMF content. An increase of IMF was observed when dietary protein or lysine were reduced (without modifying lysine or protein content, respectively) in a fatter but not in a leaner genotype. It can be concluded that modification of IMF content through the diet depends on the genotype, and that changes in dietary protein and lysine levels elicit the greatest response.

The purpose of this thesis was to determine the combined effects of protein, creatine, and conjugated linoleic acid (CLA) supplementation during resistance training. These nutritional supplements are popular during resistance training and we wanted to determine if they would have additive effects for improving body composition and strength. Forty-four participants (32 males, 12 females, mean age 20y) were randomized into three groups to receive: 1) 36 g/d protein (PRO), 2) protein and 9 g/d creatine (PRO/CR), or 3) protein, creatine and 6 g/d CLA (PRO/CR/CLA) for 5 weeks while resistance training on a four-day cycle (three days of resistance training, followed by one day of rest). Measurements at pre- and post-testing included body composition, muscle thickness of the elbow and knee flexors and extensors, and bench and leg press strength. There were time main effects (p<0.01) for strength, and muscle thickness. The PRO/CR/CLA group had significant increases in knee extensor muscle thickness over time compared to the other groups (p<0.05). There were no other differences between groups over time. The combinations of creatine and protein, or creatine, protein and CLA had no effects on body composition. It is concluded that combining protein, creatine, and CLA has minimal effects on muscular strength, muscle thickness, and body composition.

Two different studies were conducted to reduce the overall amount of omega-6 fatty acids in broiler chickens. The first experiment was performed to determine the effects of dietary conjugated linoleic acid (CLA) and omega-3 fatty acid combination on the omega-6 fatty acid accumulation in broiler chicken breast and thigh meat. Eight broilers from each treatment were processed at 4 and 6 weeks of age, respectively. Regarding the diets containing five different fat sources, broiler chickens fed CLA and fish oil diet had a lower C20:4 (arachidonic acid, AA, n-6) deposition but showed a higher n-3/n-6 ratio in breast and thigh meat than those fed a flaxseed oil diet and CLA and flaxseed oil diet (P < 0.05). The C20:4 and n-3/n-6 ratio of breast and thigh samples from fish oil diet was similar to those of the conjugated linoleic acid and fish oil combination diet (P > 0.05). However, the addition of CLA and fish oil to the diet resulted in a increase of polyunsaturated fatty acid (PUFA) concentration in broiler chicken breast and thigh meat when compared to that of fish oil diet (P<0.05). The second experiment was conducted based on six different combination of n-3 and n-9 fatty acids. One bird per pen was processed, and each bird was weighed, and blood, liver, breast and thigh samples from the bird were collected. Although the generation of prostaglandin E2 (PGE2) was not affected due to combination of n-3 and n-9 fatty acids in our diets, the deposition of n-6 fatty acids including C18:2 and C20:4 was decreased in broiler chicken breast and/or thigh muscles as n-3 fatty acids were supplied to broiler chickens for 9 weeks. Eicosapentaenoic acid (C20:5, EPA, n-3) addition to poultry diet (FEO) did not reduce the deposition of C18:2 and/or C20:4 as much as C22:6 (FDO) did. When C20:5 and C22:6 were blended to poultry diet (FHO) and fed to broiler chickens for 9 weeks, synergistic effects were observed. Reduction of C20:4 was obtained when FHO diet was fed to broiler chickens, and it may be induced due to decreased expression of delta-6 desaturase mRNA.

We hypothesized that supplementation of dietary conjugated linoleic acid (CLA) and arginine singly or in combination would increase animal performance and meat quality by decreasing adiposity and increasing lean mass in growing-finishing pigs. Sixteen pigs (80 kg) were assigned to four treatments in a 2 x 2 factorial design, differing in dietary fatty acid and amino acid composition [control: 2.05% alanine (isonitrogenous control) plus 1% canola oil (lipid control); CLA: 2.05% alanine + 1% CLA; arginine: 1% arginine + 1% canola oil; arginine + CLA: 1% arginine + 1 CLA]. Preliminary tests indicated that up to 2% arginine was acceptable without interfering with lysine absorption. Pigs were allowed to feed free choice until reaching 110 kg. There were no significant differences across treatments in feed intake, weight gain, or feed efficiency. CLA tended to decrease carcass length (P = 0.06), whereas backfat thickness tended to be greater in pigs supplemented with arginine (P = 0.08). Arginine decreased muscle pH at 45 min postmortem (P = 0.001) and tended to increase lightness of muscle at 24 h postmortem (P = 0.07). CLA supplementation increased the concentrations of trans-isomers of 18:1 (P = 0.001) and SFA (P = 0.01) in s.c. and r.p. adipose tissue. CLA supplementation increased palmitate incorporation into total lipids in longissimus muscle (P = 0.01). Glucose oxidation to CO₂ in r.p. and s.c. adipose tissue were greater in pigs supplemented with CLA in the absence or presence of arginine (P = 0.03 and P = 0.04, respectively). The volume of s.c. adipocytes in s.c. and r.p. adipose tissues was greater in pigs supplemented with CLA, arginine, or CLA plus arginine than in control pigs (P = 0.001). Neither CLA nor arginine affected the expression of PGC-1[alpha],AMPK, mTOR, CPT-1A, FAS, or SCD (P > 0.05) in any tissues. We conclude that there was no significant interaction between arginine and CLA. Supplementary CLA or arginine to finishing-growing pigs did not modulate growth performance and did not reduce adiposity. CLA increased intramuscular fat content without deteriorating meat quality traits and increased saturated fatty acids and substrate oxidation in adipose tissues. In the presence of 1% of canola oil or CLA in the diet, arginine has the potential to deteriorate meat quality by reducing early postmortem pH and by increasing carcass fatness.

A study was performed to investigate the effect of diet modifications on skin and hair coat condition in canines. The study included 24 normal adult dogs fed a baseline diet (Ol'Roy[trademark]), during an acclimation period of 12 wk (Phase I). Nine female Beagles and 15 male Hound mix-breed dogs were used. For the next 12 wk (Phase II) the dogs were divided into three groups and fed one of three specially formulated diets. They contained similar ingredients and had similar nutrient profiles except for the following differences: Diet A contained lower but adequate amounts of dietary zinc and linoleic acid than diet B. Diet C was similar to B with respect to zinc and linoleic acid but contained more α-linolenic acid. An evaluation panel conducted skin and hair coat condition scoring on wk 0, 4, 7, and 12 (Phase I) and wk 14, 16, 19, and 24 (Phase II). The panel evaluated the dogs for glossiness, softness, scale, greasiness, and overall condition. Transepidermal water loss (TEWL) and skin hydration (HYDR) assessments were determined on wk 3, 7, and 11 (Phase I) and wk 11, 12, 15, 19, and 23 (Phase II) using a Tewameter[trademark] and Corneometer[trademark] respectively. Blood samples were collected on d 0, 5, 8, 16, 28, 56, and 84. Profiles of plasma phospholipid fatty acids were determined at each collection period. Serum zinc concentrations were analyzed on wk 12, 14, and 24. The hypothesis was that a diet containing increased LA, ALA, and zinc concentrations (diet C) would show improvements of skin and hair coat condition in dogs compared to the other diets. All three test diets caused significant improvements compared to Ol'Roy[trademark]. Diet B caused more improvement than diet A in both subjective and objective assessments of skin and hair coat. Based on mean values diet B is better to be fed to dogs that need to improve skin hydration and diet C should be fed to dogs that need to decrease TEWL. Diet C not only led to improvements in skin and hair coat condition, but also provided additional benefit by producing less pro-inflammatory conditions in the skin.