abstract: The omega-3 fatty acids in fatty fish and fish oil, eicosapentanoic acid (EPA) and docosahexanoic acid (DHA), have been associated with a reduction in risk for cardiovascular disease. Blood type is a known contributor to risk for cardiovascular events. This study evaluated the effect of fish oil supplements on cardiovascular risk markers in adults with blood types A or O. An 8-week parallel-arm, randomized, double-blind trial was conducted in healthy adult men and women with either blood type A (BTA) or blood type O (BTO). Participants were randomized to receive fish oil supplements (n=10 [3 BTA/7 BTO]; 2 g [containing 1.2 g EPA+DHA]/d) or a coconut oil supplement (n=7 [3 BTA/4 BTO]; 2 g/d). Markers that were examined included total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), high-sensitivity C-reactive protein (hsCRP), and hemoglobin A1C (HbA1C). Results indicated that the percent change in LDL cholesterol was significantly greater in the coconut oil group vs the fish oil group (-14.8±12.2% vs +2.8±18.9% respectively, p=0.048). There were no other significant differences between treatment groups, or between blood types A and O, for the other cardiovascular risk markers. Further research with a larger and more diverse sample may yield a more conclusive result. / Dissertation/Thesis / M.S. Nutrition 2014

Ultraviolet radiation (UVR) is a complete carcinogen, inducing skin cancer via DNA photodamage that can lead to mutagenesis, and promoting its growth via photoimmunosuppression (PI). The omega-3 polyunsaturated fatty acid (n-3 PUFA) eicosapentaenoic acid was shown in murine studies to protect against PI and UV-induced skin cancer although the mechanism is uncertain. The principal objectives of this thesis were to (i) examine whether n-3 PUFA can protect against a clinical model of PI in healthy humans and (ii) explore whether the underlying mechanism could be abrogation of UV-induced depletion of antigen-presenting Langerhans cells (LC) from the epidermis, and/or impact on immunomodulatory cytokines. Nickel (Ni) allergic females (n=79) were randomized to 3 months of daily supplementation with 5g n-3 PUFA (70% eicosapentaenoic acid, EPA; 10% docosahexaenoic acid, DHA) or the placebo medium chain triglyceride, GTCC. Local PI was clinically assessed post supplementation using the nickel contact hypersensitivity (Ni CHS) model. In each volunteer, Ni patches were applied to 3 skin sites that were irradiated for 3 consecutive days with UV-doses of 1.89, 3.82 & 7.59J/cm2 respectively. CHS responses were measured and compared to responses of control patches applied on unirradiated skin using a reflectance erythema meter. In the same subjects, assessments of cellular and biochemical mediators of PI were made pre and post supplementation. At 24hr post irradiation with an erythemal UV-dose (4 minimal erythemal doses) to upper buttock skin, half the subjects (n=39) had skin punch biopsies taken and the other half (n=40) had suction blisters raised on this irradiated skin and on unirradiated skin of the contralateral buttock. Epidermal sheets were prepared from the punch biopsies and immunohistochemically stained to assess UV-induced LC numbers. Levels of immunomodulatory cytokines were analysed in the suction blister fluid using Luminex multiplex assay kits. To evaluate compliance and bioavailability, blood samples were taken from all volunteers, pre and post supplementation and EPA% weight in red blood cell membranes was examined using gas chromatography. Post supplementation, EPA %wt was significantly higher in the active group compared to control: mean 3.61% ± 0.22% (SEM) vs. 0.93% ± 0.06% (p<0.001). 3 volunteers showed evidence of non-compliance and were excluded from further analysis. Compared to placebo, evidence for protection against local PI of Ni CHS was apparent post n-3 PUFA at all UV doses, reaching statistical significance at the UV-dose of 3.8J/cm2 (p<0.05). No significant difference in post-UV epidermal LC numbers after supplementation was seen between active and placebo groups, with a % fall following UV of 76.61 ± 3.39% (SEM) in the active group and 73.52 ± 5.24% (SEM) in the control group. When intragroup comparisons were made pre vs. post supplementation, a similar increase in UV-induced LC depletion from the epidermis was seen in both groups, reaching statistical significance following n-3 PUFA (p=0.018). Levels of interleukins IL-10 and IL-8, and of TNF-α, increased post-UVR in both active and control groups pre-supplementation, with no changes occurring following supplementation. In conclusion, supplemental EPA was bioavailable and evidence of protection against clinical PI of Ni CHS was seen in the actively treated group. However, no evidence was found that this abrogation of PI was mediated through a reduced effect of UV on migration of epidermal LC or the immunomodulatory cytokines examined. This original study gives the first evidence that dietary n-3 PUFA may protect against clinical PI, and potentially skin cancer promotion, in humans. Further research is needed to confirm this finding, and to examine the underlying mechanisms, which could involve other immunoregulatory cells of the skin, such as dermal dendritic cells and T regulatory cells and other mediators of UV-immunosuppression including the prostanoids, which may be modified by n-3 PUFA.

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.

Omega-3 Fortification of Marinara Sauce Hannah Collie, Mary Andreae, MS, RD, LDN, W. Andrew Clark, PhD, RD, LDN, Department of Rehabilitative Science, College of Clinical and Rehabilitative Health East Tennessee State University, Johnson City, Tennessee In westernized culture, there is a deficit of healthy fats in the average person’s diet. This is evidenced by many different conditions such as cardiovascular disease, cancer, and chronic inflammatory issues. The “Mediterranean Diet” has been shown as an ideal way to combat these health issues.The diet promotes fish as a protein source and a way to consume essential polyunsaturated fatty acids. Due to location and trends, fish is less often a main component of the western diet.This study investigated ways to fortify a more commonly consumed food in western culture, marinara sauce, with flaxseed oil, walnut oil, and anchovies. These fortified marinara sauces were compared to a commercial sauce, Paul Newman’s Marinara for nutritional content. We hypothesize that adding omega-3 rich ingredients to a base marinara sauce recipe will significantly vary the fatty acid profile and increase the amount of omega-3 fatty acids as well as decrease the omega-6 to omega-3 fatty acid ratio. Sauces were prepared and analyzed for nutrient content using proximate analysis methods. Preparation included cooking and freeze drying the variants. Nutrient content tests performed included: bomb calorimetry, Kjeldahl protein analysis, Soxhlet fat analysis, ash inorganic analysis, FRAP Assay for antioxidant content, and gas chromatography to characterize fatty acid composition. Each variant sauce had a fatty acid profile that was unique. Two of the three variants showed a better omega-6 to omega-3 fatty acid ratio than the (control) Paul Newman’s sauce. Samples with the highest to lowest omega-6 to omega-3 ratio were respectively, flaxseed oil, Paul Newman’s commercial sauce, walnut oil, and anchovy. From gas chromatography, omega three fatty acid composition as a percentage of total fatty acids were approximately in the flaxseed oil variant, 21% in the Paul Newman’s sauce, 30% in the walnut oil variant, and 81% in the anchovy variant. Two of three variant sauces, walnut and anchovy, when compared to the commercial Paul Newman's sauce, showed more favorable omega-3 fatty acid content and lower omega-6 to omega-3 fatty acid ratios. Unexpectedly, the flaxseed oil variant had less omega three fatty acids and a higher omega-6 to omega-3 fatty acid ratio than the commercial sauce. In conclusion, simple additions of omega-3 ingredients to marinara sauce can decrease the omega-6 to omega-3 fatty acid ratio in the diet.

<p>Mild-moderate equine asthma (EA) is a commonly encountered disease of racehorses that affects performance. Decreasing dust exposure is crucial in the treatment of equine asthma. Dry hay, because of its high dust content, is known to increase the risk of airway inflammation. Feeding haylage, steamed hay, or hay pellets instead can help to decrease dust exposure. Haylage may also contribute to the resolution of airway inflammation by providing higher levels of omega-3 polyunsaturated fatty acids (Ω-3). Higher levels of Ω-3 are associated with an increase in pro-resolving lipid mediators (PRLM), essential molecules in the resolution of airway inflammation. </p> <p> The studies presented in this dissertation were designed to test the hypothesis that feeding low-dust forages would decrease airway inflammation in racehorses, and that haylage would provide superior resolution of airway inflammation compared to other low dust forages due to changes in systemic Ω-3 and PRLM concentrations associated with increased dietary Ω-3 intake.</p> <p>Three clinical trials were conducted to determine the effect of low-dust forages on airway inflammation. Environmental exposures were measured at the breathing zone, and bronchoalveolar lavage (BAL), and differential cytology counts were performed as measure of airway inflammation. </p> <p>The first clinical trial was a pilot study performed with 7 Standardbred racehorses. Horses were randomly assigned to eat hay (n=3) or haylage (n=4) for 6 weeks while in training. Measurements were performed at baseline and after 2, 4, and 6 weeks. Results showed a decrease in respirable dust, and β-glucan exposure in the horses fed haylage when compared to those fed hay. BAL neutrophil proportion was significantly lower at weeks 2, 4, and 6 when compared to baseline and at week 6 when compared to horses fed hay. </p> <p> The second clinical trial was performed on 19 mild asthmatic horses from the teaching herd. The diet of these horses was changed from dry hay to haylage (n=9) or hay pellets (n=10) for 6 weeks. Measurements were performed at baseline, week 3, and week 6. Results indicated that horses eating haylage and hay pellets were exposed to similar dust levels that were significantly lower than when they were eating dry hay. BAL neutrophil proportion was significantly lower in horses eating haylage when compared to baseline and to horses eating hay pellets at week 6. Horses eating haylage exhibited a significant decrease in stearic acid concentration at week 6. Pro-resolving lipid mediators (Resolvin D1, Resolvin E1, and Lipoxin A₄) did not affect neutrophil apoptosis or efferocytosis <i>in vitro</i>.</p> <p>The third clinical trial was performed on 73 thoroughbred racehorses actively racing and training. Horses were randomly assigned to eat dry hay or to change the forage to steamed hay or haylage. No other change in the management of the horses was allowed. Measurements were performed at baseline (n=73), week 3 (n=69), and week 6 (n=53). Results indicated that respirable dust exposure was significantly reduced when racehorses were fed steamed hay or haylage in place of dry hay. Respirable dust exposure was positively associated with BAL neutrophil proportions. Feeding haylage also significantly decreased exposure to respirable endotoxins. Horses eating haylage for 3 weeks had significantly lower BALF neutrophil proportion when compared with baseline. Also, at week 3, horses eating haylage showed a significant decrease in mast cell proportion, and horses eating steamed hay had a significant decrease in eosinophil proportion. At week 6, horses eating haylage maintained significantly lower BALF neutrophil proportions compared to baseline, and horses eating hay for 6 weeks. Concentration of Ω-3 and PRLM were not increased in horses eating haylage when compared to horses fed hay. Eicosapentaenoic acid was significantly higher on the horses eating haylage when compared to horses eating steamed hay. </p> <p>In conclusion, feeding low-dust forages is sufficient to decrease breathing zone exposure of horses to respirable dust. Despite similar dust exposure, haylage was the only low-dust forage that resulted in resolution of neutrophilic airway inflammation; however, the mechanism remains unclear.</p>

Thesis (D.Sc.N.S.)--Boston University, Henry M. Goldman School of Graduate Dentistry, 1993 (Nutritional Sciences). / Includes bibliography (leaves 176-187) / This dissertation has been presented in two related studies: A. Fish Oil Reduces Proteinuria and Interstitial Injury but not GIomerulosclerosis in the Milan Nomotensive Rat Rats of the Milan Normotensive strain (MNS) spontaneously develop severe Proteinuria and excessive glomemlar thromboxane (Tx)A2 PrOduction at a young age. These are accompanied by podocyte alterations and progressive focal glomerulosclerosis (FGS) and interstitial fibrosis. Since previous studies showed that pharmacologic... [TRUNCATED]

In westernized culture, there is a deficit of healthy fats in the average person’s diet. Decreased intake of omega-3 fatty acids has been correlated to many different conditions such as cardiovascular disease, cancer, and chronic inflammatory issues. The “Mediterranean Diet” has been proposed as an ideal way to combat these issues. This diet promotes fish as a protein source and as a way to increase intake of essential polyunsaturated fatty acids. Due to location and dietary trends, fish is less often the main component of Northeast Tennessee's diet. This study investigated ways to fortify a more commonly consumed food in western culture, marinara sauce, with flaxseed oil, walnut oil, or anchovies. These fortified marinara sauces were compared to a commercial sauce, Paul Newman’s Marinara. We hypothesized that adding omega-3 rich ingredients to a base marinara sauce recipe will significantly vary the fatty acid profile and increase the amount of omega-3 fatty acids and decrease the omega-6 to omega-3 fatty acid ratio. Marinara sauces were prepared and analyzed using proximate analysis methods to evaluate each variant sauce for macronutrient distribution. Preparation included cooking, freeze-drying, and grinding the variants into a fine powder. The tests that we performed included: bomb calorimetry, Kjeldahl protein analysis, Soxhlet fat analysis, ash/mineral analysis, FRAP Assay of antioxidant content, and gas chromatography to characterize fatty acid profiles. The focus of our proximate analysis was investigating the fatty acid composition to answer our main research question. Other tests conducted allowed us to understand better the nutrient composition of the 3 variants for possible future studies. The three variants showed a better omega-6 to omega-3 fatty acid ratio than the Paul Newman’s sauce. Samples with the highest to lowest omega-6 to omega-3 ratio were Paul Newman's Marinara, Anchovies, Walnut Oil, Flaxseed Oil, respectively. Gas chromatography showed different concentrations of omega-3 fatty acids as (% area under the curve): flaxseed oil (32.42%), walnut oil (7.637%), anchovies (3.018%), and Paul Newman’s Marinara (1.599%). All three variant sauces, flaxseed oil, walnut oil, and anchovy, compared to the commercial Paul Newman's sauce, had better omega-3 fatty acid content and lower omega-6 to omega-3 ratio. In conclusion, simple additions of omega-3 ingredients to marinara sauce could decrease the omega-6 to omega-3 ratio in the diet.

The use of dietary supplements occurs in the majority of American adults >20 years old and has been reported to be on the rise by the National Health and Nutrition Examination Survey. People are not only taking more dietary supplements, but taking them without the advice of a health care provider. With the lack of education time and focus on this topic, physicians need to know some evidence regarding the most common supplements used, how to understand dietary supplement labels, and where to find reputable information about dietary supplements. Vitamin D, Omega-3, and Glucosamine/ Chondroitin are reviewed here. Also, a description of the various components of a dietary supplement label is explained. Finally, a brief description of independent companies (Consumerlabs.com®, US Pharmacopedial Convention, Natural Medicine Comprehensive Database, and Natural Standard) as well as the federally run Office of Dietary Supplements are presented.

Background: Research suggests that a low omega-3 index may contribute to the low heart rate variability and the increased risk of cardiovascular morbidity and mortality in bipolar disorders. However, so far, no intervention trial with EPA and DHA has been conducted in bipolar patients attempting to increase their heart rate variability. - Methods: 119 patients with bipolar disorder according to DSM-IV were screened, with 55 euthymic bipolar patients—owing to inclusion criteria (e.g. low omega-3 index (< 6%), SDNN < 60 ms.)—being enrolled in a randomized, double-blind, 12-week parallel study design with omega-3 fatty acids (4 capsules of 530 mg EPA, 150 mg DHA) or corn oil as a placebo, in addition to usual treatment. Heart rate variability as well as the omega-3 index were measured at baseline and at the endpoint of the study. - Results: A total of 42 patients (omega-3: n = 23, corn oil: n = 19) successfully completed the study after 12 weeks. There was a significant increase in the omega-3 index (value at endpoint minus value at baseline) in the omega-3 group compared to the corn oil group (p < 0.0001). However, there was no significant difference in the change of the SDNN (value at endpoint minus value at baseline) between the treatment groups (p = 0.22). In addition, no correlation between changes in SDNN and change in the omega-3 index could be detected in the omega-3 group (correlation coefficient = 0.02, p = 0.94) or the corn oil group (correlation coefficient = − 0.11, p = 0.91). Similarly, no significant differences between corn oil and omega-3 group regarding the change of LF (p = 0.19), HF (p = 0.34) and LF/HF ratio (p = 0.84) could be demonstrated. - Conclusions: In our randomized, controlled intervention trial in euthymic bipolar patients with a low omega-3 index and reduced heart rate variability no significant effect of omega-3 fatty acids on SDNN or frequency-domain measures HF, LF and LF/HF ratio could be detected. Possible reasons include, among others, the effect of psychotropic medication present in our trial and/or the genetics of bipolar disorder itself. Further research is needed to test these hypotheses.

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