Lipid oxidation is the main factor which limits the shelf-life of meat when held under frozen storage. Research undertaken used pork phospholipid liposomes as a model for studying lipid oxidation in meat. Oxidation was followed by monitoring the decrease in the phospholipid unsaturated fatty acyl chains. It was found that the greater the level of unsaturation of the phospholipid fatty acyl chain the greater was their susceptibility to peroxidation. However, the results were not consistent and several reasons for the variation in rate are provided. At ambient temperatures copper (II) was found to be pro-oxidant in the peroxidation of liposomes. At temperatures below 0°C the prooxidant activity of copper (II) was significantly reduced. However copper again became highly pro-oxidant if sodium chloride was present. It is suggested that salt controls the copper ion concentration at sub-zero temperatures as the pro-oxidant activity of copper (II) is reduced on increasing the copper (II) concentration from 0.9 to 90 ppm. Other experiments found sodium nitrite and pholyphosphate to act as antioxidant and that liposome structure was an important factor in the rate of peroxidation. Four storage trials on pork burgers were undertaken to determine whether salt was also pro-oxidant in the stability of pork when held under frozen storage. The oxidative deterioration of the meat was followed by the following methods of analysis:- 1. The decrease in the unsaturated acyl chains of both total lipid and phospholipid. 2. The change in the colour parameters of the meat using reflectance spectroscopy. 3. The analysis of neutral lipid oxidation products by HPLC. 4. The organoleptic qualities of the pork using a trained panel of food assessors. The results from these storage trails showed that the deterioration of pork was minimised by storing the burgers at lower temperatures within the range 0 to -30°C. Salt was found to accelerate the oxidative deterioration of both uncooked and cooked pork when stored at -20°C. Nitrite was found to exhibit some antioxidant behaviour and reduce the pro-oxidant effect of salt.

A series of experiments was conducted to test the overall hypothesis that reducing the omega-6 (n-6) to omega-3 (n-3) fatty acid (FA) ratio in sow diets would improve sow reproductive performance (characterized by increases in numbers and body weight of piglets born alive and weaned) and would lessen the inflammatory responses of their offspring post weaning. Diets were wheat/barley based and consisted of a control (tallow based, similar to a standard production diet), 3 diets with plant oil based n-6:n-3 ratios (9:1P, 5:1P, and 1:1P) and a 5:1 fish oil diet (5:1F). The control diet had a ratio of 8:1, but contained approximately half the polyunsaturated FA content of the other diets. Sows were randomly assigned to a treatment diet on d 80 of gestation, and remained on that treatment for three consecutive reproductive cycles (gestation/lactation 1 = P1, gestation/lactation 2 = P2, gestation/lactation 3 = P3). Experiment 1 was designed to test the hypothesis that reducing the n-6:n-3 FA ratio in sow diets would increase circulating concentrations of n-3 FA’s in sows and in their offspring, and the passive immune status of piglets would be improved. Performance data was collected throughout P1 and P2 on 150 sows (n = 30/diet). Sow and piglet serum, colostrum and milk were analyzed for FA profiles, and colostrum and piglet serum were analyzed for immunoglobulin (Ig) A and IgG. In P1, birth weights were unaffected by diet (P > 0.05). Average piglet weaning weights (P = 0.02) and ADG (P = 0.01) however, were highest for piglets born to sows consuming the 9:1P and 5:1P diets. During P2, 5:1F sows consumed 10% less feed (P = 0.04), their piglets had reduced birth weights (P = 0.05), and average weaning weight was reduced by 0.8 kg (P = 0.04) relative to control or 5:1P sows. Colostral and piglet plasma IgA and IgG were unaffected by diet (P > 0.05). Colostrum FA profile patterns were similar to that of the sow diets. Serum n-3 FA’s were greatest in sows (P < 0.01) and piglets (P < 0.01) consuming 1:1P or 5:1F diets. Serum α-linolenic acid (ALA) was highest in the 1:1P sows and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were highest in the 5:1F sows. In piglet serum obtained prior to suckling, ALA and DHA did not differ among treatments (P > 0.05) but EPA was 2.5 times greater in the 1:1P group and 4 times greater in the fish group (P < 0.01) compared to those from the control diet. In post-suckle samples, ALA was highest in serum from 1:1P diet piglets (P < 0.01), and EPA and DHA were highest in piglet serum from the 5:1F sows (P < 0.01). Omega-3 FA’s can perturb lipid metabolism, specifically increasing the lipolytic activity of adipose tissue and thus the second experiment tested the hypothesis that high producing sows, consuming reduced n-6:n-3 ratios would have increased body fat mobilization. Twenty sows per diet, farrowing ≥ 11 piglets and nursing ≥ 10 piglets during P3, were used. Performance data on sows and piglets (such as weights, numbers, backfat changes) was collected throughout lactation and milk samples obtained on d 4 and d 16 of lactation. Jugular catheters were inserted into 8 sows from each of the 9:1P and 1:1P groups on d 5 of lactation and sows were challenged with a single injection of epinephrine followed by serial blood collections. Feed intake was highest for sows consuming the control (8.4 kg/d) and 5:1P (8.2 kg/d) diets and lowest for the sows fed the 1:1P (7.4 kg/d) and 5:1F (7.7 kg/d) diets (P = 0.05). Altering the n-6:n-3 FA ratio did not affect sow BW, piglet ADG, milk DM and N content or the total output of milk (P > 0.2). Sows consuming the 1:1P diet had greater backfat thickness (P < 0.05) and numerically higher plasma NEFA at baseline compared with the 9:1P sows (240 vs 93 uM; P = 0.16). When given epinephrine, 9:1P fed sows tended to have lower net incremental area under the curve (niAUC) glucose (P = 0.08) and numerically higher niAUC NEFA (P = 0.17) and glycerol (P = 0.15). A third experiment was conducted to test the hypothesis that piglets raised by sows consuming reduced n-6:n-3 ratios would have reduced inflammatory responses post-weaning. Piglets (n = 20/diet) raised by sows consuming the treatment diets described above for 2 gestation/lactation cycles (P2) were selected at weaning. Within diet group, pigs were randomized to either a challenge control group (saline injected) or to a lipopolysaccharide (LPS) injected group (n=10/challenge•diet-1). Piglets were fed a common starter diet for 6 days followed by saline or LPS injections on d 7. Rectal temperatures were recorded for 24 hrs and blood samples were collected at 0, 2, 6 and 12 hrs post injection for pro-inflammatory cytokine and blood urea nitrogen (BUN) analysis. Injecting LPS caused decreased feed intake and reduced ADG (P < 0.01), and increased temperature and cytokine production (P < 0.05). Piglets raised by sows consuming the 1:1P diet had elevated temperatures (P = 0.01; diet x challenge P > 0.05). Overall, circulating plasma ALA and EPA were increased in sows and piglets when sows were fed a 1:1 plant based ratio compared to the control or high n-6:n-3 ratio groups. Sows fed a ratio of 1:1 mobilized more body fat relative to those consuming the 9:1 ratio; there were no treatment effects on piglet growth. Reducing maternal n-6:n-3 FA ratios below 5:1 increased piglet body temperature prior to and during an LPS induced inflammatory challenge,. Reducing the sow dietary n-6:n-3 FA ratio below 5:1 may have detrimental effects on piglets due to over-stimulation of inflammatory responses.

Zugl.: Hamburg, Hochsch. für Angewandte Wissenschaften, Diplomarbeit, 2004.

Thesis (M.S.)--Oregon State University, 2009. / Printout. Includes bibliographical references (leaves 104-111). Also available on the World Wide Web.

PUFAs have been shown to modulate ALD by several mechanisms, including free radical generation from hepatic lipid peroxidation. However, how they modulate lipid peroxidation and generation of bioactive metabolites in ALD is poorly understood and it is still not clear which PUFAs (ω-3 or ω-6) are beneficial or detrimental in ALD. Thus, our objective was to study the effect of ω-3/ω-6 PUFAs on lipid peroxidation and ethanol mediated steatosis and inflammation. Using standard liquid diet (LDC), LDC with fish oil (rich in ω-3) and safflower oil (rich in ω-6), we studied the generation of bioactive metabolites, such as eicosanoids and free radicals generated via lipid peroxidation. In addition, we determined the effect of PUFAs on several inflammatory and fibrotic factors, e.g. gene as well as protein expression, using western blot and RT-PCR, respectively. We also investigated the effect of PUFA diets on novel targets, such as hepatic membrane transporters with potential role in liver inflammation. Our results suggest that ω-3 diet prevented while ω-6 based diets promoted the development of fatty liver and inflammation. ω-3 PUFA reduced AA-peroxidation by lowering hepatic AA concentration and expression of peroxidation enzymes, COX-2 and 5-LOX, resulting in lower generation of pro-inflammatory AA-derived PGs (Series-2), HETEs and free radicals, along with increase in anti-inflammatory EPA and DHA-derived PGs (Series-3). ω-3 diet might also reduce liver inflammation by preventing activation of NF-кB and induction of TNF-α. Rats fed with ω-3 diet showed high protein expression of efflux transporters, MRP-2 and ABCA1, indicating elimination of peroxidation metabolites and triglycerides from the liver and decreased inflammation. In contrast, ω-6 diets led to increase in AA-peroxidation and generation of AA-derived pro-inflammatory metabolites. ω-6 based diets also promoted fatty liver and inflammation by activating NF-кB, inducing TNF-α and downregulation of efflux transporters, MRP-2 and ABCA1. This study not only provides new insights into the effects and possible mechanisms by which ω-3 and ω-6 PUFAs may alter hepatic steatosis and inflammation, but also put forward new targets of research, such as hepatic membrane transporters in relation to liver pathology in ALD.

Reproductive efficiency in beef cattle can be improved with reproductive technologies at the herd, individual cow, and embryonic levels. Decreasing the bull:cow ratio for natural service after fixed time artificial insemination (FTAI) can alleviate economic burden associated with FTAI. In experiment 1, the total number of cows exposed per bull was negatively correlated with pregnancy rate to natural service on first return to estrus after FTAI in fall herds. The number of open cows per bull in fall herds using one natural service sire was negatively correlated with pregnancy rate on first return to estrus. There was no correlation between number of cows exposed per bull and pregnancy rates in fall herds with multiple sires or in spring herds. However, bull:cow ratio accounted for only 5–11% of the variation in pregnancy rates, thus we conclude that a reduced bull:cow ratio did not affect natural service return to estrus pregnancy rate. Experiment 2 examined how supplementing calcium salts of soybean oil (CSSO) improves beef cow fertility. Non-pregnant cows received supplement with either saturated fat or omega-6 rich CSSO. There were no changes in dominant follicle diameter, corpus luteum volume, plasma progesterone, or endometrial gene expression (PTGES and AK1B1, PPARA, PPARA, PPARD) between treatments. Plasma and follicular fluid fatty acid compositions were altered between treatments. Experiment 3 examined if size parameters of zygotes have potential as a non-invasive, objective embryo selection method. The outer diameter, area of ooplasm, and thickness of zona pellucida (ZP) was digitally measured on individual artificially activated oocytes and in-vitro fertilized (IVF) zygotes. Larger outer diameter increased probability of development to the blastocyst stage by days 7 and 8 for activated oocytes and tended to by day 8 for IVF zygotes. Thinner ZP increased probability of development to blastocyst stage on days 7 and 8 for oocytes, and to day 8 for IVF zygotes. Area did not affect development but was positively correlated with blastomere number on day 8. An interaction between diameter and ZP thickness was observed in zygotes, but not activated oocytes, suggesting oocyte activation is not always a suitable replacement for in-vitro fertilization. / Doctor of Philosophy / We need to improve reproductive efficiency in beef cows if we are to combat the challenges of producing more food while using less resources due to limited land availability and concerns with greenhouse gas emissions from agriculture. In cow-calf production systems, this means producing one healthy calf per cow per year. Cattlemen can implement a variety of assisted reproductive technologies to achieve this goal. Achieving maximal reproductive efficiency will require using technologies that are incorporated into herd management, individual animal care, and in vitro embryo production. Fixed time artificial insemination (FTAI) allows cattlemen to maximize the number of cows becoming pregnant and calving earlier in the season to increase efficiency. Unfortunately, use of FTAI is uncommon in cow-calf production systems because of labor and economic restraints. In order to improve economic feasibility of FTAI, bull-related costs need to be reduced, which can be done through increasing the number of cows serviced per bull (decreasing the bull:cow ratio). This study retrospectively examined correlations between the bull:cow ratio and pregnancy rate on first return to estrus after FTAI. There was little to no correlations between bull:cow ratio and pregnancy rates, and if they were significant, there was much variation in the data. With this we concluded that a reduced bull:cow ratio does not affect pregnancy rate on first return to estrus, allowing producers to increase the number of cows serviced by a single bull and reduce bull related costs. Supplementing calcium salts of soybean oil (CSSO) that are rich in omega-6 fatty acids can enhance beef cow fertility, but it is unclear why this happens. Using non-pregnant cows as a model, we studied the effects of either saturated fat or CSSO on reproductive parameters such as ovarian structures, hormone concentrations, and uterine gene expression. There were no changes in any of these parameters between treatments, but there were changes in the concentrations of certain plasma and follicular fluid fatty acids. There was also reduced activity of lipid metabolism enzymes. We were unable to pinpoint how CSSO supplementation improves reproduction, but the altered fatty acid content of tissues and altered enzyme activity likely plays a key role, thus ultimately impacting fatty acid utilization and growth of the embryo. In-vitro embryo production can increase the number of offspring produced from a single female and accelerate the incorporation of animals with high genetic merit into herds. To obtain optimal pregnancy rates with in vitro embryos, we should develop non-invasive, objective methods for identifying the most viable embryos. This study examined if size parameters of activated and fertilized oocytes are indicative of successful development. We discovered that oocytes with large diameters and those with thin zona pellucida were most likely to develop to the blastocyst stage, and that the area of the cell was positively correlated with blastocyst total cell number. An interaction between diameter and ZP thickness was observed in zygotes, but not activated oocytes, suggesting oocyte activation is not always a suitable replacement for in-vitro fertilization. This suggests that digital measurements of fertilized oocytes may have potential as objective selection criteria. Addressing issues of reproductive inefficiency in beef cows requires a comprehensive approach, as there is not one ideal solution. Management techniques can alleviate the cost of FTAI by reducing the number of bulls used without affecting pregnancy rates. Supplementing CSSO can alter tissue fatty acids to enhance fertility. Finally, the efficiency of in vitro embryo production can be improved by selecting better embryos for transfer without compromising the embryo. Combinations of all these techniques can create more reproductively efficient animals.

O experimento foi realizado no Biotério Experimental da USP em Pirassununga - SP. Na primeira fase foram utilizados 42 coelhos machos em crescimento da raça Nova Zelândia mantidos individualmente durante todo o experimento. O ensaio biológico totalizou 150 dias. Na Fase 1 os animais receberam dietas com indução de hipercolesterolemia (0,5% de colesterol), com desequilíbrio na proporção de Omega-6/Omega-3 (n-6:n-3) de 15:1 e vice versa e balanço na proporção de n-6:n-3 de 4:1 (controle), totalizando 6 ensaios, os quais foram divididos aleatoriamente em 7 grupos de 6 coelhos cada. Óleos de girassol e de peixe foram utilizados como fontes de ácidos graxos poli-insaturados n-6 e n-3, respectivamente. Na Fase 2, os animais remanescentes continuaram recebendo as mesmas dietas, exceto no grupo B (hipercolesterolemia) que passou a receber o tratamento controle para verificar o efeito. No final da Fase 1, três animais de cada grupo foram eutanasiados com retirada da artéria aorta para determinação de placas lipídicas, histopatológica, colesterol total e imunohistoquímica para verificação da expressão da enzima LDL-receptor e determinação histopatológica do tecido hepático. Em soro foram realizadas determinações de perfil de ácidos graxos, colesterol total, LDL e HDL-colesterol e triacilgliceróis. Células endoteliais aórticas de coelhos foram isoladas para a realização do teste dose resposta com óleo de peixe para identificar a melhor relação do consumo destes ácidos graxos comparando com o experimento in vivo. O objetivo deste trabalho foi monitorar os efeitos do fornecimento de cada dieta sobre o teor de colesterol total, LDLcolesterol, HDL-colesterol e triacilgliceróis em sangue de coelhos. No geral, o perfil de ácidos graxos no soro correspondeu diretamente com a dieta consumida. Os principais efeitos dos ácidos graxos poli-insaturados Omega 3 foi a redução dos lipídios séricos quando os coelhos remanescentes da dieta hipercolesterolêmica (0,5% de colesterol e n-6:n-3 de 2:1) passaram a receber o equilíbrio da relação de ácidos graxos 4:1 de n-6:n-3. O teor de colesterol total no soro, artérias e o crescimento dos ateromas foram influenciados pelo elevado consumo de ácidos graxos n-6 e n-3 associados à adição de colesterol nas dietas. Por outro lado verificamos que o excesso de Omega 3 associado ou não ao colesterol contribuiu para o agravamento das placas ateroscleróticas inclusive deposição de cálcio nas mesmas e paredes endoteliais. Provavelmente, o excesso de n-3 tenha ocasionado efeito contrário às suas funções anti-inflamatória, antiagregatória e anti-trombótica. Dietas hipercolesterolêmicas provocaram esteatose hepática e o elevado consumo de Omega-6 em detrimento do baixo consumo de Omega -3 induziram hepatite crônica. Já o consumo elevado de Omega-3, sem colesterol, regrediu a esteatose hepática nos animais. O consumo equilibrado de n-6:n-3 reduziu a expressão da enzima LDL-receptor no grupo que anteriormente recebeu dieta hipercolesterolêmica. Ressaltando-se a importância do consumo equilibrado destes ácidos graxos já que esta enzima é controlada pelo colesterol livre circulante. No teste dose-resposta em células endoteliais da aorta de coelhos foi evidenciado que a melhor proporção de n-6:n-3 seria de 9 vezes mais do que a concentração de Omega 3 recomendada de 5:1. / The experiment was conducted at the Experimental Biotery by USP in Pirassununga - SP. In the first phase were used 42 male rabbits growing New Zealand breed individually maintained throughout the experiment. The biological assay amounted to 150 days. In phase 1, the animals received diets induced hypercholesterolemia (0.5% cholesterol) with imbalance in the ratio of Omega-6 / Omega-3 (n-6 / n-3) 15:1 and vice versa, and balance the ratio of n-6 / n-3 of 4:1 (control), totaling six tests, which were randomly divided into 7 groups of six rabbits each. Sunflower and fish oils were used as sources of fatty acids polyunsaturated n-6 and n-3, respectively. In Phase 2, the remaining animals continued to receive the same diets except in group B (hypercholesterolemia) who went on to receive the control treatment to check the effect. At the end of Phase 1, three animals from each group were sacrificed with the aorta artery removed for determination of lipid plaques, histopathology, immunohistochemistry and total cholesterol to verify the expression of LDL-receptor enzyme and determination of histopathological liver tissue. In serum profile were made determinations of fatty acids, total cholesterol, LDL and HDL cholesterol and triglycerides. Aortic endothelial cells of rabbits were isolated to perform the dose response test with fish oil to identify the best value for the consumption of these fatty acids compared to the in vivo experiment. The objective of this work was to monitor the effects of provision for each diet on total cholesterol, LDL-cholesterol, HDLcholesterol and triglycerides in the blood of rabbits. In general, the profile of fatty acids in serum corresponded directly with the diet consumed. The main effects of polyunsaturated Omega 3 fatty acids was the reduction of serum lipids when the remaining rabbits with hypercholesterolemic diet (0.5% cholesterol and n-6: n-3 2:1) have received the balance of the relationship fatty acid 4:1 n-6: n-3. The total cholesterol content in serum, the growth of arteries and atheroma were influenced by a high intake of n-6 fatty acids and n-3 associated with the addition of cholesterol in diets. On the other hand we verified that the excess of Omega 3 with or without cholesterol contributed to the aggravation of the atherosclerotic plaques including deposition of calcium in them and endothelial walls. Probably the excess of n-3 has caused the opposite effect to their anti-inflammatory, anti-thrombotic and anti-aggregatory functions. Hypercholesterolemic diets caused hepatic steatosis and high intake of Omega-6 to the detriment of low consumption of Omega -3 induced chronic hepatitis. High consumption of Omega-3, no cholesterol, decreased hepatic steatosis in animals. Highlighting the importance of the balanced intake of this fatty acids, since this enzyme is controlled by circulating free cholesterol. In dose-response test in endothelial cells of the rabbits aorta was evident that the best ratio of n-6: n-3, nine times more than the recommended concentration of Omega 3 to 5:1.

O experimento foi realizado no Biotério Experimental da USP em Pirassununga - SP. Na primeira fase foram utilizados 42 coelhos machos em crescimento da raça Nova Zelândia mantidos individualmente durante todo o experimento. O ensaio biológico totalizou 150 dias. Na Fase 1 os animais receberam dietas com indução de hipercolesterolemia (0,5% de colesterol), com desequilíbrio na proporção de Omega-6/Omega-3 (n-6:n-3) de 15:1 e vice versa e balanço na proporção de n-6:n-3 de 4:1 (controle), totalizando 6 ensaios, os quais foram divididos aleatoriamente em 7 grupos de 6 coelhos cada. Óleos de girassol e de peixe foram utilizados como fontes de ácidos graxos poli-insaturados n-6 e n-3, respectivamente. Na Fase 2, os animais remanescentes continuaram recebendo as mesmas dietas, exceto no grupo B (hipercolesterolemia) que passou a receber o tratamento controle para verificar o efeito. No final da Fase 1, três animais de cada grupo foram eutanasiados com retirada da artéria aorta para determinação de placas lipídicas, histopatológica, colesterol total e imunohistoquímica para verificação da expressão da enzima LDL-receptor e determinação histopatológica do tecido hepático. Em soro foram realizadas determinações de perfil de ácidos graxos, colesterol total, LDL e HDL-colesterol e triacilgliceróis. Células endoteliais aórticas de coelhos foram isoladas para a realização do teste dose resposta com óleo de peixe para identificar a melhor relação do consumo destes ácidos graxos comparando com o experimento in vivo. O objetivo deste trabalho foi monitorar os efeitos do fornecimento de cada dieta sobre o teor de colesterol total, LDLcolesterol, HDL-colesterol e triacilgliceróis em sangue de coelhos. No geral, o perfil de ácidos graxos no soro correspondeu diretamente com a dieta consumida. Os principais efeitos dos ácidos graxos poli-insaturados Omega 3 foi a redução dos lipídios séricos quando os coelhos remanescentes da dieta hipercolesterolêmica (0,5% de colesterol e n-6:n-3 de 2:1) passaram a receber o equilíbrio da relação de ácidos graxos 4:1 de n-6:n-3. O teor de colesterol total no soro, artérias e o crescimento dos ateromas foram influenciados pelo elevado consumo de ácidos graxos n-6 e n-3 associados à adição de colesterol nas dietas. Por outro lado verificamos que o excesso de Omega 3 associado ou não ao colesterol contribuiu para o agravamento das placas ateroscleróticas inclusive deposição de cálcio nas mesmas e paredes endoteliais. Provavelmente, o excesso de n-3 tenha ocasionado efeito contrário às suas funções anti-inflamatória, antiagregatória e anti-trombótica. Dietas hipercolesterolêmicas provocaram esteatose hepática e o elevado consumo de Omega-6 em detrimento do baixo consumo de Omega -3 induziram hepatite crônica. Já o consumo elevado de Omega-3, sem colesterol, regrediu a esteatose hepática nos animais. O consumo equilibrado de n-6:n-3 reduziu a expressão da enzima LDL-receptor no grupo que anteriormente recebeu dieta hipercolesterolêmica. Ressaltando-se a importância do consumo equilibrado destes ácidos graxos já que esta enzima é controlada pelo colesterol livre circulante. No teste dose-resposta em células endoteliais da aorta de coelhos foi evidenciado que a melhor proporção de n-6:n-3 seria de 9 vezes mais do que a concentração de Omega 3 recomendada de 5:1. / The experiment was conducted at the Experimental Biotery by USP in Pirassununga - SP. In the first phase were used 42 male rabbits growing New Zealand breed individually maintained throughout the experiment. The biological assay amounted to 150 days. In phase 1, the animals received diets induced hypercholesterolemia (0.5% cholesterol) with imbalance in the ratio of Omega-6 / Omega-3 (n-6 / n-3) 15:1 and vice versa, and balance the ratio of n-6 / n-3 of 4:1 (control), totaling six tests, which were randomly divided into 7 groups of six rabbits each. Sunflower and fish oils were used as sources of fatty acids polyunsaturated n-6 and n-3, respectively. In Phase 2, the remaining animals continued to receive the same diets except in group B (hypercholesterolemia) who went on to receive the control treatment to check the effect. At the end of Phase 1, three animals from each group were sacrificed with the aorta artery removed for determination of lipid plaques, histopathology, immunohistochemistry and total cholesterol to verify the expression of LDL-receptor enzyme and determination of histopathological liver tissue. In serum profile were made determinations of fatty acids, total cholesterol, LDL and HDL cholesterol and triglycerides. Aortic endothelial cells of rabbits were isolated to perform the dose response test with fish oil to identify the best value for the consumption of these fatty acids compared to the in vivo experiment. The objective of this work was to monitor the effects of provision for each diet on total cholesterol, LDL-cholesterol, HDLcholesterol and triglycerides in the blood of rabbits. In general, the profile of fatty acids in serum corresponded directly with the diet consumed. The main effects of polyunsaturated Omega 3 fatty acids was the reduction of serum lipids when the remaining rabbits with hypercholesterolemic diet (0.5% cholesterol and n-6: n-3 2:1) have received the balance of the relationship fatty acid 4:1 n-6: n-3. The total cholesterol content in serum, the growth of arteries and atheroma were influenced by a high intake of n-6 fatty acids and n-3 associated with the addition of cholesterol in diets. On the other hand we verified that the excess of Omega 3 with or without cholesterol contributed to the aggravation of the atherosclerotic plaques including deposition of calcium in them and endothelial walls. Probably the excess of n-3 has caused the opposite effect to their anti-inflammatory, anti-thrombotic and anti-aggregatory functions. Hypercholesterolemic diets caused hepatic steatosis and high intake of Omega-6 to the detriment of low consumption of Omega -3 induced chronic hepatitis. High consumption of Omega-3, no cholesterol, decreased hepatic steatosis in animals. Highlighting the importance of the balanced intake of this fatty acids, since this enzyme is controlled by circulating free cholesterol. In dose-response test in endothelial cells of the rabbits aorta was evident that the best ratio of n-6: n-3, nine times more than the recommended concentration of Omega 3 to 5:1.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Monika Novotná Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Title of Diploma Thesis: The determination of fatty acid levels in the tissues of healthy, suddenly deceased persons and polymorbid patients The aim of the thesis was to determine the levels of 14 fatty acids in tissues of suddenly deceased, otherwise healthy individuals and in the group of polymorbid, chronic patients. It was a comparison of fatty acid levels in seven tissues of the human body: subendocardial left ventricular tissue, liver parenchyma tissue, kidney cortex, adrenal tissue, skeletal muscle, abdominal subcutaneous adipose tissue, and brain tissue. Each group included 10 deceased patients. The theoretical part incudes fatty acids as the main component of lipids. It deals mainly with the group of polyunsaturated fatty acids and their relation to pathologies in the human body. Gas chromatography, thanks to which we analyzed the fatty acid spectrum, is also described. The experimental part consists of the basic characteristics of the research groups, the description of the workflow and the results. Statistically processed data are divided into categories by individual fatty acids and other monitored...

Introduktion: Torra ögon (DES) och Sjögrens syndrom (SjS) är inflammatoriska sjukdomar som drabbar ögonen. DES drabbar tårkörtelns funktionella del vilket ger ögonsmärta och suddig syn. SjS är en kronisk autoimmun sjukdom som ger skada på tår- och salivproducerande körtlar. Det leder till kliniska symtom i form av mun- och ögontorrhet. Essentiella fettsyror bildar proinflammatoriska och antiinflammatoriska cytokiner som kan hjälpa vid behandling av dessa sjukdomar. Metod och syfte: En litteraturstudie genomfördes för att ta reda på om fleromättade fetter kan lindra symtomen vid SjS och DES. Resultat: Resultaten för studierna visade resultat på OSDI, Schirmers test, BUT, IL-17, tårosmolaritet, tårvolym, ostimulerat salivflöde, stimulerat salivflöde, sonderingsdjup, PGE1, van Bijstervelds poäng och flourscein infärgning. Diskussion: Resultatet visar signifikanta skillnader för vissa doser av n-3 och n-6. Det som skiljer resultaten åt är de olika doserna av fettsyror samt vad dess placebokontroller intog. OSDI visade främst skillnader på de patienter med DES men endast på en grupp med SjS som intog n-3 för behandling av torr mun. Schirmers test och BUT visade en ökning hos patienterna med mild och måttlig DES men inte på patienterna med SjS. Patienterna med SjS visade förbättringar på stimulerat salivflöde och OSDI för torr mun samt PGE1 nivåer och flourscein infärgning samtidigt som en del studier inte visade några signifikanta förbättringar på några mätvariabler. Det kan grunda sig i intaget av fettsyror, sjukdomstillstånd eller längden på behandling. Slutsats: En del personer kan få förbättrade symtom av att inta n-3 eller n-6 tillskott men skillnaderna är inte alltid statistiskt signifikanta för studierna. / Introduction: Dry eye syndrome (DES) and Sjögren ́s syndrome (SjS) are inflammatory diseases that affect the eyes. DES affects the lacrimal glands functional unit, causing eye pain and blurred vision. SjS is a chronical autoimmune disease that causes damage to tear and salivary glands. It leads to clinical symptoms in the form of mouth and eye irritation. Essential fatty acids form pro-inflammatory and anti-inflammatory cytokines that can help in the treatment of these diseases. Method and purpose: A literature study was performed to study if essential fatty acids can relieve the symptoms of DES and SjS. Results: The results for the studies showed results on OSDI, Schirmer ́s test, BUT, IL- 17, tear osmolarity, tear volume, unstimulated salivary flow, stimulated salivary flow, depth of probing, van Bijsterveld ́s score and flourscein staining. Discussion: The result shows significant differences for certain doses of n-3 and n-6. What separates the results are the different doses of fatty acids and what their placebo controls took. OSDI showed major differences in patients with DES but only in one group of patients with SjS who took n-3 for treating dry mouth. Schirmer ́s test and BUT showed an increase in patients with mild and moderate DES but not in patients with SjS. Patients with SjS showed improvements in stimulated salivary flow, dry mouth OSDI, PGE1 levels and flourscein staining, while some studies did not show any significant improvements in any measurement variables. It may be due to the intake of fatty acids, disease states or the length of treatment. Conclusion: Some people may get improved symptoms of taking n- 3 or n-6 supplements, but the differences are not always statistically significant for the studies.