HT biomass sonicated at ES inputs < 56KJ/gTS decreased floc size by 41% and enhanced its metabolic activity by 50-250% compared to control. ES inputs >118 KJ/gTS caused HT biomass solubilization and irreversible loss of its metabolic activity and reflocculation ability. During continuous activated sludge processing (ASP) of real primary effluent the observed yield (Yobs) decreased by 20% compared to control ASP at SF (stress factor) of 1 (biomass exchanged without USPT). At SF of 0.5, 1 and 1.5 (biomass exchanged with USPT) the Yobs further decreased by 33, 25 and 44% respectively as compared to control. This indicated that combining biomass anoxic exposure with USPT enhanced sludge reduction by increasing microbial maintenance metabolism likely in combination with microbial flora shift in the ASP depending on SF.

The thesis contains a theoretical overview, which has focused on classification of lipids, fatty acids and their metabolisms, with emphasis on polyunsaturated fatty acids inclusive of their conversion to eicosanoids, functions and effects of eicosanoids on human health. Furthermore, the theoretical part of the thesis has focused on a food’s enrichment of polysaturated fatty acids n-3 and several possibilities of analytical determination of fatty acids. The practical part of the thesis has aimed attention at the determination of fatty acids in the liver, fat and muscles’ tissue of the experimental rats. The animals were fed with a standard feed mixture, which contains 5 % of fish oil, 5 % of palm oil, 5 % of safflower oil and 5 % of Schizochytrium algae extract. The obtained fat was extracted from the lyophilized samples followed by derivatization and analytical determination of the fatty acid derivatives by gas chromatography. From the results of the experimental part could be determined, that the addition of 5 % of fish oil to the feed mixture of the experimental animals increased the deposition of EPA and DHA in all examined tissues (p < 0.05). The addition of 5 % of the Schizochytrium extract to the animal’s feed mixture had an impact on the effective deposition of DHA and DTA in the examined tissues of all used oils (p < 0.05). Both of the previously mentioned oils can be considered as the most effective nutraceuticals for reducing the ratio of PUFA n-6/n-3 in the food. Unlike the addition of 5 % of safflower or 5 % of palm oil (as a control), ratios of PUFA n-6/n-3 were many times higher. Therefore, from the healthy point of view, this can be concluded as inappropriate.

Ovarian cancer is the fifth most lethal cancer in women (1) and the most lethal gynecological malignancy. In 2018, there were approximately 22,240 new diagnosed cases of ovarian cancer and 14,070 deaths in the United States alone (2). The lifetime risk for developing ovarian cancer in the United States is 1.3% or approximately 1 in 78 women. The five-year survival rate for women with ovarian cancer is a grim 47.6% (2) while the average five year survival rate for all cancers is about 68%. This dismal prognosis for ovarian cancer patients indicates the critical need for improved treatment options, efficient early detection methods and effective preventative measures for ovarian cancer (1). The objective of this study was to determine if DHA causes a reduction in cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) by blocking the activation of NF-κB regulated transcription in the ovary. DHA is a 22 carbon long-chain omega-3 polyunsaturated fatty acid that is biologically derived from Alpha-linolenic acid (ALA) found in flaxseed. COX-2 is an enzyme that catalyzes the conversion of arachidonic acid to prostaglandins. Prostaglandin E2 (PGE2) is a key regulator of inflammation which has been shown to be highly associated with ovarian cancer development and progression. Our laboratory studies ovarian cancer in the laying hen because it is the only known animal model to naturally develop ovarian cancer that both pathologically and histologically matches that of the human form of the disease. Dietary flaxseed is one of the richest vegetable sources of omega-3 polyunsaturated fatty acids. Our previous studies have shown that in laying hens, a long-term flaxseed supplemented diet reduces the incidence and severity of ovarian cancer and decreases COX-2 and PGE2. It was hypothesized that DHA, derived from ALA found in flaxseed, decreases inflammation in the ovaries by suppressing the activation of COX-2 and the production of PGE2 through inhibition of the NF-κB pathway. For this study, an NF-κB reporter plasmid was transfected into HEK293 cells. The reporter plasmid (“met-luc”) produces a secreted luciferase allowing sequential analysis of media from DHA and TNF-α treated cells to assess changes in NF-κB transcriptional activation. Tumor necrosis factor alpha (TNF-α)-induced activation of NF-κB was used as a positive control. NF-κB activation was also assessed by measuring its nuclear translocation and cytoplasmic accumulation through immunocytochemistry (ICC) and western blot analysis. In a parallel study, immortalized ovarian surface epithelial (IOSE) cells were challenged with the same treatments of DHA and TNF-α. In these cells, COX-2 mRNA was assessed through RT-qPCR and COX-2 protein expression was analyzed through ICC and western blot.Our results indicate that DHA acts in a cell specific manner to reduce inflammation associated with cancer. We have found that in HEK293 cells DHA reduces TNFα induced NF-κB reporter activity. In contrast, ALA does not affect NF-κB reporter activity. HEK293 cells treated with TNFα alone indicated a dose-dependent increasing trend in nuclear translocation of the NF-κB p65 subunit and a decreasing trend in cytoplasmic p65, suggesting potential increased pathway activation. ICC suggests DHA treatment causes increased cytoplasmic sequestration of the NF-κB p65 subunits indicating inhibition of TNFα induced NF-κB activation. Western blot data also indicates a decreasing trend in nuclear NFκB p65 when cells are pretreated with DHA and subsequently challenged with TNF. The IOSE cells, were the only cells out of the cell lines tested (BG1, HEYC2, TOV112D, SKOV3, HEK293) to express COX-2. In these IOSE cells, TNFα alone showed a dose-dependent increasing trend in COX-2 protein (analyzed through ICC and western blot) and mRNA levels (analyzed through RT-qPCR). ICC analysis revealed that DHA reduces TNF induced COX-2 protein expression. However, the western blot did not further support this observation. Only a slight non-significant reduction with DHA treatment was observed. In addition, both DHA and TNFα, while also not significant, seemed to increase mRNA levels of COX-2 compared to control. This slight decreasing trend in COX-2 protein expression and increase in mRNA, could indicate a possible post-transcriptional mechanism of regulation of COX-2 by DHA independent of NF-κB in the IOSE cells. These data suggest that DHA could act via distinct mechanisms in a cell specific manner to potentially reduce COX-2 and subsequently PGE2 levels. DHA can act at the transcriptional level by reducing the nuclear translocation of NF-κB and transcriptional activation of NF-κB target genes such as COX-2 in some cell types. DHA also has the potential to work via a post-transcriptional mechanism to inhibit COX-2 and in turn reduce PGE2 levels. Both mechanisms ultimately have the potential to decrease the inflammation associated with ovarian cancer. This study describes the anti-inflammatory action of dietary flaxseed consumption, making flaxseed supplementation a promising preventive measure for reducing the risk of ovarian carcinogenesis.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第18343号 / 農博第2068号 / 新制||農||1024(附属図書館) / 学位論文||H26||N4850(農学部図書室) / 31201 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 植田 充美, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Inflammation is a complex physiological response normally initiated by the innate immune system, often as a response to exposure to otherwise harmful stimuli. While generally useful in humans as a protective response to foreign matter, chronically elevated quantities of associated inflammatory factors C-reactive protein, TNF-alpha, IL-6, and IL-1beta have been linked in literature with decreased overall lifespan and well-being in humans via inflammatory processes. It is possible that by lowering these associated factors, increased well-being and lifespan may be experienced by the general population. One common health supplement with such promise is fish oil, which, through compounds eicosapentaenoic acid and docosahexaenoic acid, has been observed to decrease levels of secreted inflammatory markers in cell culture. In addition, molecular pathways have since been discovered which demonstrate possible means for which this physiological response may occur. However, despite the promise of such health benefits, studies attempting to discern the impact EPA/DHA supplementation has on inflammatory markers within humans have since emerged with mixed results. The aim of this study is to provide a meta-analysis across a number of studies to determine whether or not an impact exists through EPA/DHA supplementation in healthy populations, and if one exists, to what degree the respective inflammatory factors may be lowered.

No description available.

Pacific bluefin tuna (PBT; Thunnus orientalis) has grown significantly in popularity in recent years due to the globalization of Japanese cuisine. PBT is highly sought after for sushi and sashimi products due to its great quality and taste. Wild populations of this species have been affected by their increasing popularity, pushing innovators in the food industry to create meat alternative versions of PBT. The muscle composition of PBT varies, leading to different types (cuts) of meat in a way that is analogous to various cuts of beef. This study evaluated the differentiation amongst the 6 distinct cuts, including otoro, ventral akami, dorsal akami, ventral chu-toro, dorsal chu-toro, and wakaremi conducting volatile analysis, proximate analysis, and fatty acid analysis. The results from these analyses can then be used as a base standard for companies seeking to create alternatives versions of PBT. Samples analyzed in this study were cultured PBT species that were caught as juveniles and raised in captivity on a PBT farm in Mexico. Volatile analysis was conducted using a SPME GC/MS method. Overall, 41 aroma compounds were identified in PBT that met the identification criteria, including 9 aldehydes, 7 alcohols, 14 alkanes, 2 ketones, 4 alkenes, 3 aromatic compounds, and 2 miscellaneous compounds. Proximate analyses were conducted using standard methods. Significant differences (p <0.05) were found between each cut for the proximate analysis. The fatty acid analysis determined that there were twenty-two identifiable fatty acids found in the different cuts. The omega-3 fatty acids eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) with DHA being present at a higher amount than EPA in each cut. Overall, there are similarities and differences among the different cuts of bluefin tuna that researchers would need to mimic to provide adequate nutritional and sensorial properties of PBT. / Master of Science in Life Sciences / Pacific bluefin tuna (PBT; Thunnus orientalis) is a fish that has gained tremendous popularity over the years due to the globalization of Japanese cuisine. This tuna species is synonymous with high quality and great taste, making it key for sushi and sashimi dishes. The increased demand for this species has caused wild populations to decrease; therefore, the food industry has sought to create meat alternatives for the species. The PBT has 6 distinct cuts that make up the composition of the fish, similar to the differences that can be found in beef cuts. These 6 distinct cuts are otoro, ventral akami, dorsal akami, ventral chu-toro, dorsal chu-toro, and wakaremi. The purpose of this study is to determine the aroma composition, fatty acid composition, and fat, moisture, ash, and protein contents of the different cuts. To measure the aroma composition, gas-chromatography mass spectroscopy (GC-MS) was used; it is a machine that can be used to identify and measure the aroma compounds of products. Standard procedures were used for the other analyses. Overall, 41 aroma compounds were identified in PBT that met the identification criteria, consisting of aldehydes, alcohols, alkanes, ketones, alkenes, aromatic compounds, and some miscellaneous compounds. The cuts displayed distinguishable differences in their fat, ash, moisture, and protein contents. The fatty acid analysis concluded that there were twenty-two identifiable fatty acids found in the different cuts. In this study, close attention was paid to omega-3 fatty acids due to their health benefits for consumers. The omega-3 fatty acids eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) were found in all cuts with DHA being present at a higher amount than EPA in each cut. From this study, researchers have the foundation for understanding the composition of PBT to create a meat alternative that meets consumer expectations.

Foram utilizadas 168 galinhas poedeiras Hisex White em delineamento experimental inteiramente casualizado, com duração de cinco semanas. As aves foram distribuídas em sete tratamentos com três repetições cada, com o objetivo de verificar a influência da suplementação de três fontes marinhas de PUFAs n-3 - óleos de salmão (SA) e de atum e sardinha (A/S) e mistura de algas marinhas (AL), bem como sua combinações (SA+A/S, AL+SA, AL+A/S) - em relação ao grupo controle alimentado com ração basal de milho e soja (CON), sobre a qualidade do ovo e a composição lipídica da gema. A qualidade externa e interna do ovo não foi alterada significativamente pela adição de fontes de PUFAs n-3 na dieta das aves. A relação entre os lípides saturados : monoinsturados : poliinsaturados da gema (3,5 : 4,5 : 2,0) manteve-se constante com a adição dos suplementos de PUFAs n-3 na ração. Os óleos de salmão (SA) e de atum e sardinha (A/S) revelaram-se mais efetivos no enriquecimento da gema do ovo em PUFAs n-3, em especial o DHA. A relação PUFAs n-6/ PUFAs n-3 sofreu redução significativa com a adição das fontes marinhas de ômega-3 à dieta das galinhas. O EPA, embora em teores mais baixos que o DHA na gema do ovo, apresentou incremento significativo com o uso dos óleos de peixes (SA e A/S) adicionado à dieta em relação ao CON. O consumo de dois ovos enriquecidos com DHA do presente estudo, por dia, provavelmente atenderia os requerimentos diários deste PUFA n-3 para o ser humano / To investigate the influence of three PUFAs n-3 marine sources - salmon oil (SA), tuna and sardine oil (A/S), marine algae (AL) and combinations (SA+A/S, AL+SA, AL+A/S) - on egg quality and egg lipid composition, compared to a corn/soy control group (CON), 168 Hisex White laying hens were assigned into seven treatments with three repetitions in a randomized design during a experimental period of five weeks. The external and internal egg quality were not significantly affected by the addition of PUFAs n-3 marine sources into the hen diets. The saturated : monounsaturated : polyunsaturated ratio (3.5:4.5:2.0) remained constant by the supplementation of the marine sources into the diets. The salmon oil and the tuna and sardine oil showed the best results of PUFAs n-3 egg yolk enrichment, mainly the DHA. The PUFAs n-6/PUFAs n-3 ratio was significantly reduced by the inclusion of PUFAs n-3 sources into the hen diets. The EPA, although with lower levels than DHA into the egg yolk, showed a significant increase by the use of salmon oil and tuna and sardine oil added to the diet as compared to the control group (CON). The consumption of two DHA enriched eggs from this study per day probably would reach the daily requirements of this PUFA n-3 for the human been

INTRODUÇÃO: Apesar de classicamente a lipoproteína de alta densidade (HDL) ser descrita como um fator de risco independente para as doenças cardiovasculares (DCV), devido ao seu papel no transporte reverso de colesterol (TRC), atualmente sabe-se que esta partícula exerce múltiplos mecanismos biológicos que podem contribuir para esta cardioproteção, com estudos indo além do conteúdo de colesterol associada à HDL (HDL-C). OBJETIVO: Avaliar o impacto da suplementação de ?-3 no perfil de ácidos graxos, tamanho e capacidade antioxidante da HDL. MÉTODOS: Estudo clínico, aleatorizado, controlado, duplo cego e paralelo, onde foram incluídos 147 indivíduos adultos, com fatores de risco cardiovascular, que foram alocados no grupo ?-3 (n=77 - 3,0 g/dia contendo 1,8 g de EPA+DHA) ou no grupo ?-6 (n=70 - 3,0 g/dia de óleo de girassol contendo 1,95 g de ácido linoleico). No início (T0) e após 8 semanas (T8) amostras de sangue foram coletadas e, a partir do plasma ou soro, foram analisados o perfil lipídico (CT, HDL-C, LDL-C, TG), as apolipoproteínas AI, CII e CIII, as subfrações da HDL (Lipoprint®), a atividade da paraoxonase (PON1) e da proteína transportadora de éster de colesterol (CETP), a capacidade antioxidante da HDL (método experimental) e o perfil de ácidos graxos e conteúdo de AGNEs da HDL. Os resultados do efeito do tempo, da intervenção e das interações entre os parâmetros monitorados e os desfechos foram realizados com o auxílio do programa SPSS® versão 20.0. O valor de significância considerado foi de p<0,05. RESULTADOS: Os grupos ?-3 e ?-6 eram semelhantes entre si em relação a idade, sexo, tabagismo e uso de medicamentos, ambos apresentando alta prevalência de hipertensão e dislipidemia. Nos dois grupos, houve redução em todos os marcadores lipídicos, exceto a concentração de HDL-C, que aumentou. Observou-se o efeito do tempo no conteúdo de HDLAGNEs (?=-16,2%), no percentual de HDLGRANDE (?=20,1%) e de HDLPEQUENA (?=-5,0%). O aumento de EPA na HDL se associou à menor chance de ter elevação da atividade da PON1 (OR=0,446; IC=0,200-0,994), da concentração de HDLAGNEs (OR=0,275; IC=0,113-0,660) e do percentual de HDLPEQUENA (OR=0,337; IC=0,146-0,782). Observou-se ainda que o EPA se associou a, aproximadamente, 3,5 mais chances de aumento no percentual da HDLGRANDE (OR=3,522; IC=1,652-7,507). Quanto ao aumento de DHA na HDL, este esteve associado de maneira significativa à diminuição da concentração de Apo AI (OR=0,351; IC=0,150-0,821), além da atividade da PON1 (OR=0,226; IC=0,110-0,639) e da concentração de HDLAGNEs (OR=0,275; IC=0,113-0,668). Resultados similares aos obtidos com o EPA foram observados para o DHA e o tamanho da HDL. Não foi constatado efeito da incorporação de EPA e DHA na resistência à oxidação. CONCLUSÃO: A intervenção com ?-3 promoveu mudanças na composição da partícula de HDL, aumentando o percentual das subfrações maiores, sem, contudo, modificar sua capacidade antioxidante. / BACKGROUND: Although high-density lipoprotein (HDL) is classically described as an independent risk factor for cardiovascular disease (CVD), because of its role in reverse cholesterol transport (RCT), currently, it is known that this particle exerts multiple biological mechanisms that may contribute to this cardioprotection, with studies going beyond HDL cholesterol content (HDL-C). OBJECTIVE: To evaluate the impact of ?-3 supplementation on the fatty acid profile, size and antioxidant capacity of HDL. METHODS: A randomized, controlled, double-blind, parallel clinical study involving 147 adult subjects with cardiovascular risk factors, who were allocated into the ?-3 group (n=77 -3.0 g/day containing 1.8 g EPA + DHA) or the ?-6 group (n=70 -3.0 g/day of sunflower oil containing 1.95 g of linoleic acid). In the beginning (T0) and after 8 weeks (T8) blood samples were collected and, from plasma or serum, were analyzed lipid profile (TC, HDL-C, LDL-C, TG), apolipoproteins AI, CII and CIII, HDL subfractions (Lipoprint®), paraoxonase (PON1) and cholesterol ester carrier protein (CETP) activities, antioxidant capacity of HDL (experimental method), the fatty acid profile and NEFAs content HDL. Results of the effect of time, intervention and interactions between monitored parameters and outcomes were performed with the aid of SPSS® software version 20.0. The significance level considered was p<0.05. RESULTS: The ?-3 and ?-6 groups were similar in relation to age, sex, smoking and medication use, both presenting high prevalence of hypertension and dyslipidemia. In both groups, there was a reduction in all lipid markers except HDL-C concentration, which increased. The effect of time on HDLNEFAs content (?=-16.2%), and on the percentage of HDLLARGE (?=20.1%) and HDLSMALL (?=-5.0%) were observed. Increasing EPA in HDL was associated with a lower chance of enhance PON1 activity (OR=0.446, CI=0.200-0.994), HDLNEFAs concentration (OR=0.275, CI=0.113-0.660), and HDLSMALL (OR=0.337, CI=0.146- 0.782). It was also observed that EPA was associated with approximately 3.5 more chances of raising the percentage of HDLLARGE (OR=3.522, CI=1.652-7.507). Increasing DHA in HDL was significantly associated with a decrease in Apo AI concentration (OR=0.351, CI=0.150- 0.821), and also PON1 activity (OR=0.226, CI=0.110-0.639) and concentration of HDLNEFAs (OR=0.275, CI=0.113-0.668). Similar results obtained with EPA were observed for DHA and HDL size. No effect of the incorporation of EPA and DHA on oxidation resistance was observed. CONCLUSION: The ?-3 intervention promoted changes in the composition of the HDL particle, increasing the percentage of the larger subfractions, without, however, modifying its antioxidant capacity.

Alimentos e suplementos adicionados de ácidos graxos poliinsaturados ômega 3, tais como os ácidos eicosapentaenóico (EPA) e docosaexaenóico (DHA), tem sido comercializados com a alegação funcional de reduzir as concentrações de triacilgliceróis (TG) e, assim, reduzir o risco para doenças cardiovasculares. Entretanto, pelo fato desses ácidos graxos apresentarem uma cadeia carbônica altamente insaturada, o consumo crônico em doses elevadas poderia levar a um aumento da susceptibilidade do organismo ao estresse oxidativo. O desequilíbrio nos processos oxidativos pode ser um agravante em indivíduos dislipidêmicos, uma vez que essa população normalmente controla as altas concentrações de lipoproteína de baixa densidade (LDL) com medicamentos como as estatinas, e a oxidação da LDL é um dos fatores mais importantes para o desenvolvimento da aterosclerose. Uma vez que a proposta dos alimentos funcionais e suplementos nutracêuticos é de agir como coadjuvantes à prescrição terapêutica, indivíduos sob tratamento a base de estatinas seriam um público-alvo para produtos adicionados de ácidos graxos ômega 3. Desta forma, a proposta deste estudo foi de avaliar o efeito de uma suplementação contendo EPA e DHA sobre alguns biomarcadores do estresse oxidativo em mulheres com dislipidemia controlada por estatinas. Seguindo delineamento crossover duplo cego, 45 indivíduos foram distribuídos em 4 grupos e receberam a seguinte suplementação: cápsulas de óleo de milho/soja, sem estatina (Placebo); cápsulas de milho/soja, com estatina (Estatina); cápsulas de óleo de peixe, sem estatina (Omega 3); cápsulas de óleo de peixe, com estatina (Combinado). Os indivíduos foram orientados a consumir 4 cápsulas (2,3 g de EPA e DHA) por dia durante 42 dias. Foram realizadas coletas de sangue antes e após o período de intervenção para analise das concentrações séricas de glicose, lipoproteínas, perfil de ácidos graxos no plasma e marcadores do estresse oxidativo. A suplementação com EPA e DHA na dosagem de 2,3 g/dia foi efetiva na redução das concentrações séricas de LDL e TG apenas na presença de estatinas, confirmando o uso combinado desses compostos para melhoria do perfil lipoprotéico. Por outro lado, essa suplementação parece induzir um aumento do estresse oxidativo, evidenciado pelo aumento da concentração plasmática de malondialdeído, provavelmente decorrente da redução da expressão da catalase promovida pelos AGPI n-3. / Food and supplements that contain n-3 fatty acids, such as eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, have been commercialized using functional claim to reduce triacylglycerol (TG) concentration and, hence, reduce the risk of cardiovascular diseases. However, as these fatty acids are highly unsaturated, the chronic consumption in high doses could raise the susceptibility of the organism to oxidative stress. Oxidative processes imbalance can be a problem in dyslipidemic individuals, once this population usually controls the high concentration of low density lipoproteins (LDL) with drugs such as statins, and oxidation of LDL is one of the most important factors to the atherosclerosis development. Since functional foods and nutraceutical supplements are supposed to contribute to the therapeutic prescription, dyslipidemic individual under statins treatment could be the target to products containing n-3 fatty acids. Thus, the objective of this study was to evaluate the effect of a supplementation containing EPA and DHA on some biomarkers of oxidative stress in women with dyslipidemia controlled by statins. A randomized double-blind placebo-controlled crossover study was carried out with 45 women who were distributed into 4 groups that received the following supplementation: corn/soy oil capsules, no statin (Placebo); corn/soy oil capsules, with statin (Statin); fish oil capsules, no statin (Omega 3); fish oil capsules, with statin (Combined). The individuals were instructed to consume 4 capsules (2.3 g of EPA and DHA) every day during 42 days. Blood samples were collected before and after the supplementation to evaluate serum glucose and lipoprotein concentrations, plasma fatty acid profile and biomarkers of oxidative stress. N-3 PUFA supplementation (2.3 g/day) was effective in reducing serum LDL and TG concentrations only when combined with statins, confirming the combined use of these compounds for improvement of lipoprotein profile. On the other hand, this supplementation seemed to induce an increase of the oxidative stress, evidenced by the higher malondialdehyde plasma concentrations, probably due to the reduction in catalase expression promoted by n-3 PUFA.