Dissertations / Theses on the topic 'Blood fatty acid'

Conversion of the essential n-3 (18:3n-3) and n-6 (18:2n-6) fatty acids to longer chain polyunsaturated fatty acids (PUFA) involves sequential desaturation and elongation reactions. Previous studies have reported gender differences in n-3 PUFA synthesis, whereas the effect of age is less clear. n-3 PUFAs are reported to have important effects on immune cell function. A previous study reported long chain PUFA synthesis in mitogen stimulated but not quiescent peripheral blood mononuclear cells (PBMCs). However, the underlying mechanism is not known. PUFA synthesis was investigated in PBMCs incubated with [1-13C]18:3n-3 for 48 h. Activation with the T-lymphocyte mitogen concanavalin A (Con A) increased PUFA synthesis. 22:6n-3 synthesis was not detected. [1-13C] incorporation was greatest for 20:3n-3 suggesting initial chain elongation is an important fate for 18:3n-3. Con A increased expression of three key genes (FADS2, FADS1 and ELOVL5) involved in PUFA synthesis, suggesting upregulation of the pathway is controlled at the transcriptional level. ELOVL2 expression was negligible, possibly explaining the lack of 22:6n-3 synthesis. Con A increased methylation of 12 CpGs in the FADS2 promoter contradicting the general view that DNA methylation represses transcription. Subsequent 5’RACE analysis verified that activated PBMCs were not using an alternative promoter for FADS2 transcription. Contrary to expectation, 18:3n-3 conversion in activated PBMCs was not affected by gender or menopausal status and there was no clear age effect. PUFA synthesis was constitutive in the Jurkat T-lymphocyte leukaemic cell-line and was higher than in PBMCs. FADS2, FADS1 and ELOVL5 mRNA expression was also higher in Jurkat cells and was associated with 50% lower methylation of 17 CpGs in the FADS2 promoter, suggesting transcriptional dysregulation of PUFA synthesis in Jurkat cells involves altered DNA methylation. These findings have provided novel insights into the regulation of PUFA biosynthesis in PBMCs and upregulation of the pathway in activated PBMCs suggests that newly synthesised PUFAs may be important for cell function.

Red blood cells (RBC) have been shown to mediate plaque development seen in coronary artery disease (CAD). This study determined whether differences in RBC fatty acid (FA) composition were related to CAD risk. FAs were extracted from RBCs of 38 individuals who have undergone cardiac catheterization, 9 of whom had obstructive CAD, and analyzed via gas chromatography. Ferric reducing ability of plasma (FRAP) assay was used to determine oxidative stress. Food frequency questionnaires were used to correlate RBC omega-3 FA to daily intake of omega-3 FA. No correlation was found between RBC content and intake of omega-3 FA. FRAP values and RBC FA composition did not differ between the 2 groups with exception of the saturated FA, palmitic acid (p=0.018). These results suggest that RBC FA composition may differ between individuals with or at risk for CAD. Additional research is needed to validate this biomarker as a predictor of CAD.

To evaluate the relative efficacy of plant sterols (PS) esterified with the fatty acids from fish oil (PS-FO), olive oil (PS-OO) and sunflower oil (PS-SO) on blood lipid and PS concentrations, 21 hyperlipidemic subjects were randomly assigned to each of five treatments for 28 days using a cross-over design. The results showed that: (1) in a comparison of olive oil (OO), fish oil (FO), PS-FO and PS-SO subgroup, PS-FO reduced triacylglycerols (TG) relative to PS-SO. Total cholesterol (T-C)/high-density lipoprotein cholesterol (HDL-C) ratio was reduced with PS-FO compared with FO. Plasma PS levels were increased with PS-SO and PS-FO. (2) in a comparison of OO, PS-SO and PS-OO subgroup, PS-OO had a larger decrease in T-C than OO, while PS-SO and OO reduced T-C equally. Both PS-SO and PS-OO elevated plasma PS levels. Overall, PS-FO and PS-OO have a higher potential for decreasing the risk of cardiovascular disease in hyperlipidemic subjects than PS-SO and OO supplementations.

Background: The traditional Omani diet of dates, milk, rice, brown bread, fish and vegetables has undergone considerable change, now resembling a Western diet that is high in calories, high glycaemic index carbohydrates, total fat and saturated, trans and omega 6 fatty acids, and low in omega-3 fatty acids and essential micronutrients. The available data on daily food intake and its impact on micro- and macro-nutrients are scanty in the Arab world. Obtaining these data is especially important in light of the growing prevalence of non-communicable diseases (NCDs) - cardiovascular diseases, diabetes, cancer and respiratory diseases - which now account for more than 60% of the global disease burden and mortality, and contribute to more than 50% of annual deaths in Arabian countries, including Oman. Moreover, obesity, an antecedent of NCDs, has reached epidemic proportions in the region. Since NCDs develop gradually during the course of the lifespan, various national and international committees on NCDs have recommended that children and young adults be the primary focus of any action plan that aims to prevent and control non-communicable diseases. Specific aims: The aims of the study are: a) To assess the nutrient intake, body composition, blood lipids, blood pressure, blood glucose and cognitive behaviour of Omani school children; b) To assess the level of Vitamin A, D, E, and Beta Carotene among the Omani school children c) To assess the level of red blood cell fatty acids of Omani school children d) To investigate the effect of fish consumption on red blood cell omega-3 fatty acids and plasma fat-soluble vitamins; e) To investigate the impact of omega-3 fatty acids obtained from fish oil capsule on the red blood cell omega-3 fatty acids and plasma fat-soluble vitamins. Methods: Children ages 9 and 10 years (n = 314) were recruited from three randomly selected schools in the Muscat Governorate. The schools were assigned to a fish, fish oil, or control group and the children were accordingly provided a lightly grilled oily fish, a re-esterified triacylglycerol fish oil capsule, or no fish for 12 weeks. Baseline body weight, height and body mass index were assessed and a non-fasting blood sample collected at baseline and after 12 weeks of intervention. Data on food intake were gathered using a 24-hour recall questionnaire; also collected were data on weight, height, blood pressure, triglycerides and fasting glucose. In addition, teachers completed the National Initiative for Children’s Health Quality Vanderbilt Assessment Scales-Teacher Assessment Scale to assess the prevalence and sub-types of ADHD. Results: At baseline, a significant number of the children had low levels of vitamin D, and omega-3 fatty acids. The low levels of vitamin D and omega-3 fatty acids were ameliorated significantly by fish consumption and fish oil supplementation. In terms of weight, the prevalence of overweight or obesity for the total sample was 28.2% (10%, 46%) and 22.6% (8%, 38%). That is, about one fourth of the study subjects were overweight or obese. With regards to ADHD, the prevalence rate of PIS, PHIS and ADHDCIH among Omani school children was 7.3%, 3% and 8.8%, respectively. After 12 weeks intervention, the children who received fish oil (54.1±17.5 nmol/L; p < 0.001) and fish (49.2±17.4 nmol/L; p < 0.05) had elevated levels of vitamin D than those who did not (42.3±17.5 nmol/L). The fish oil (1.2±0.70 μmol/L) and fish (1.20±0.7 μmol/L) groups also had higher concentrations of beta carotene than the placebo group (0.85±0.43 μmol/L; p < 0.0001). Systolic and diastolic blood pressure (p=0.0001) and plasma triglycerides (p < 0.05) but not HDL, LDL and total cholesterol (p > 0.05) were lower in the fish oil group compared with those who were fed oily fish. Conclusions: This study provides important and underreported data on nutrient intake levels by school children in Arab world. For the first time, these data will be available as a benchmark for future research and health programs in the region. A large number of boys and girls had diets that failed to provide the recommended levels of daily nutrients. This study also indicates that ADHD is relatively common among Omani school children; additional studies are needed to assess the generalisability of these findings. Vitamin D deficiency is prevalent in Omani school children, but can be mitigated with omega-3 fatty acid supplementation. This study also provides evidence that Omani pre-adolescents have a low level n-3 fatty acid index that can be ameliorated by fish oil supplementation or consumption of oily fish. Hence, there is a need for a child-focused program of food fortification, school feeding programmes, targeted intervention with n-3 fatty acid enriched food products, family nutrition education and outdoor activities to alleviate the problem.

The objective of the study was to determine the effects of Aloe ferox inclusion in drinking water on growth performance, blood biochemistry, physico-chemical characteristics, fatty acid profile and oxidative stability of broiler meat. The importance of A. ferox as a medicinal plant and factors that influence its utilization by communal poultry farmers were also investigated by use of a questionnaire survey. The survey revealed that the majority of respondents (84.6%) faced health challenges in their chickens and many relied (96.2%) on A. ferox to treat diseases and control parasites. The study also revealed that the choice of medicine (traditional or conventional) was influenced (P<0.05) by level of education and income. In the second phase of the research, a total of 600 Ross 308 day-old broilers, were randomly put in 6 treatment groups with 4 replicates, each having 25 birds. Fresh aqueous A. ferox leaf juice (ALJ) was administered in drinking water at a dosage of 20ml/litre to T1, T2 and T3 from day one to day 35, day one to day 14 and day 15 to day 28, respectively. Birds in T4 and T5 (positive controls) were treated with terramycin at the recommended dosage of 14g/litre of drinking water from day one to day 6 and from day 15 to 20, respectively; and birds in T6 (negative control) received distilled water from day 1 to 35. Feed Intake (FI), average daily gain (ADG) and feed conversion ratio (FCR) were calculated for the 5 week trial. After slaughter, carcass characteristics particularly dressing percentage (DP) and relative organ weight (ROW) were calculated. Serum biochemistry was also determined. For meat quality, pH and color were recorded 45 minutes and 24 hours after slaughter from the breast muscle. Fatty acid profiling and oxidative stability were determined using meat samples from the breast and thigh muscles. The results for growth performance showed that thebirds which were given A. ferox for the first two weeks (T2) consumed significantly (P>0.05) more feed (189.4g) than those in the negative control (159.6g) at the beginning of the starter phase. Subsequently, their ADG recorded on day 7 (27.1g) and day 14 (43.1g) were significantly (P<0.05) higher than the negative control (22.8g and 36.2g, respectively). Significant treatment effects (P<0.05) for FCR were reported in the 4th week for the birds that received A. ferox throughout (T1: 3.5). Carcass characteristics were not significantly (P>0.05) affected by A. ferox inclusion in drinking water. The highest high density lipoprotein (HDL) values (2.78 mmol/L) were yielded in T2and T3 had the lowest values (0.61mmol/L) for low density lipoprotein (LDL). For physico-chemical properties, no significant effects (P>0.05) of treatment on pH, colour, cooking loss and tenderness were observed. However, the group treated with A.ferox throughout the production cycle, had the highest pH (6.2), lowest lightness (38.5), highest redness (4.1), highest tenderness (13.86N) and the lowest cooking loss (12.6%). Significant treatment effects (P<0.05) were observed on the composition of the PUFA eicosatrienoic acid (C20:3c8, 11, 14(n-6)) of the breast muscle which was significantly lower in the A. ferox treatment groups than the positive controls. For the thigh muscle, there were significant (P<0.05) treatment effects on composition of palmitoleic acid (C16:1c9) and g-linolenic acid (C18:3c6, 9, 12 (n-3)). No significant (P>0.05) effects were found on oxidative stability of both thigh and breast muscles. In conclusion, the wide use of A. ferox by communal chicken farmers showed its importance as a medicinal plant. Apart from it being an effective medicinal plant, A. ferox inclusion in drinking water results in improved FI, ADG, reduced in LDLC and better g-linolenic and palmitoleic acid composition in the meat.

Ultraviolet radiation (UVR) in solar light is important for skin biology. It is involved in the development acute and chronic skin inflammation, aging and cancer, causing erythema, tanning and local or systemic immunosuppression. Omega-3 polyunsaturated fatty acids (n-3 PUFA) are considered anti- inflammatory and could reduce the damage caused by overexposure to UVR. Although, n-3 PUFA have been considered as photoprotective agents, their exact mechanisms of action is not completely understood. The aim of the work is to determine the effect of UVR and the n-3 PUFA eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) on human skin cells (in vitro study), specifically on: cell viability, apoptosis and their metabolism through the cyclooxygenase and lipoxygenase pathways. Also, to study the cellular incorporation and effect of n-3 PUFA on the fatty acid profile of skin cells. A clinical study was undertaken to assess the incorporation of n-3 PUFA supplements in human skin. A clinical study was performed in 40 healthy women (active group) supplemented with 4g/day of EPA (70%) and DHA (10%) and 40 healthy women (placebo group) supplemented with 4g/day of glyceryl tricoprylate coprate (GTCC). After 3 months, both blood samples and skin punch biopsies were collected and analysed for fatty acids by gas chromatography (GC). HaCaT keratinocytes and 46BR.1N fibroblasts were cultured and treated with 10 and 50μM of either EPA, or DHA or oleic acid (OA) for 72h and exposed to 15 and 50 mJ/cm2. Cell viability was measured by the MTT assay and cell apoptosis by a colorimetric method, at 24h post UVR. Cells and culture media were analysed by GC and liquid chromatography tandem mass spectrometry (LC/ESI-MS/MS) to assess cellular fatty acids and production of eicosanoids. The clinical a study showed that in RBC saturated fatty acids (SFA) (44.27±7.43%) were the main fatty acid group followed by n-6 PUFA (29.61±5.53%). While in dermal tissue monounsaturated fatty acids (MUFA) (58.90±9.80%) was the main fatty acid group followed by SFA (27.06±6.78%). A significant increase in EPA, DHA and docosapentaenoic acid (DPA) was observed in RBC but only EPA was significantly increased in the dermis post n-3 PUFA supplementation. . The viability of HaCaT keratinocytes and 46BR.1N fibroblasts decreased post UVR and this was further reduced post PUFA treatment. Cell apoptosis increased when cells were exposed to UVR and further increased when cells were treated with EPA and DHA. . In HaCaT keratinocytes MUFA (54.22±8.82%) was the main fatty acid group followed by FAS (37.11±.9.16%), while SFA (51.94±8.68%) was the main group followed by MUFA (27.07±4.79) in 46BR.1N. Treated both cells with EPA and DHA showed significant increased in cellular EPA, DPA and DHA. 46BR.1N fibroblasts produced higher levels of prostaglandins (PG) compared to HaCaT keratinocytes: PGE2 and PGD2 were the main PG in both HaCaT (7.96±3.18 and 1.48±1.19 pg/million cell; respectively) and 46BR.1N with (44.2±23.00 and 17.1±9.71 pg/million cell; respectively). Significant increase in PGE1 and PGE2 occurred when cells were exposed to 15mJ/cm2 UVR. Treatment with n-3 PUFA decreased the level of PGE1 and PGE2, and increase production PGE3 at the baseline and post UVR. Both cell lines produced hydroxy fatty acids and the concentration of these mediators was higher in 46BR.1N than HaCaT. The concentrations of these mediators were significant increased post UVR: treatment with n-3 PUFA decreased the level of HODE and HETE, and increase production of HEPE and HDHA at baseline and post UVR. Overall, n-3PUFA treatment led to increases in the content of EPA and DHA on RBC, dermal tissue and human skin cell lines. EPA and DHA in skin cell lines appear to offer protection by increasing cellular apoptosis, decreasing inflammatory mediators specifically PGE2 and 12-HETE, and increasing anti-inflammatory mediators such as PGE3, 15-HEPE and 17-HDHA.

Parallel increases in many inflammatory diseases including atopy over the last 40 years suggest that common environmental changes may be promoting inflammatory immune responses. Modern diets have become increasingly rich in n-6 polyunsaturated fatty acids (PUFAs) and relatively deficient in n-3 PUFAs. These dietary changes are believed to promote a pro-sensitisation, pro-allergic and pro-inflammatory environment. Exposure to such an environment during pregnancy and in the very early life period is considered to influence subsequent patterns of the immature and developing neonatal immune system, and this may contribute to the increase in allergic disease in early life. As allergic diseases often first manifest in infancy, prevention strategies need to be targeted early, even in utero. Epidemiologic and experimental data provide a plausible link between dietary changes and increased incidence of childhood atopic disease. Although there have been studies examining the potential benefits of giving n-3 PUFA-rich fish oil supplements during pregnancy, there are no studies examining the effects of increased consumption of oily fish in pregnancy on neonatal immune responses and subsequent clinical outcomes. The Salmon in Pregnancy Study (SIPS) is the first randomised controlled trial of oily fish intervention during pregnancy. The hypotheses being investigated in SIPS is that increased intake of salmon, a source of long chain (LC) n-3 PUFAs (eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)), in pregnancy will a) increase maternal LC n-3 PUFA intake, b) increase maternal and infant blood LC n-3 PUFA status, c) modulate fetal/neonatal immune responses and d) lower the risk of infant atopy determined at 6 months of age. The primary outcome measures of SIPS were the clinical signs of atopy in the offspring. Pregnant women (n=123) at high risk of having atopic offspring, and with low habitual intake of oily fish (≤ 2/month) were randomised at 20 weeks of pregnancy to either consuming 2 portions/week of farmed salmon (n=62) or continuing their habitual diet (n=61) until the end of pregnancy. The woman attended a clinic at 20 (n=123), 34 (n=110) and 38 (n=91) weeks of gestation at which fasting blood was collected and a food frequency questionnaire (FFQ) was administered (at 20 and 34 weeks). At delivery umbilical cord blood was collected (n=101) for fatty acid and immunological analysis. Infants attended a clinic at 6 months of age (n=86) for assessment of allergic sensitisation by skin prick testing (SPT) using various allergen extracts and of atopic dermatitis (SCORAD index). Maternal and cord plasma and cord blood mononuclear cell (CBMC) fatty acid compositions were determined by gas chromatography. Neonatal (cord) immune cell subsets were identified by flow cytometry. Ex-vivo cytokine production by CBMC in response to stimulants (allergen, mitogen, and toll-like receptor (TLR) ligands) was determined by cytometric bead array and flow cytometry. Ex-vivo prostaglandin E2 production by CBMC was determined by enzyme-linked immunosorbent assay. Immunoglobin E concentration was measured in cord blood plasma and in 6 month infant blood plasma. Eating oily fish twice a week during pregnancy resulted in a higher maternal intake of LC n-3 PUFAs (both EPA and DHA) and in higher maternal and cord blood plasma status of LC n-3 PUFAs (both EPA and DHA). LC n-3 PUFA content of CBMC was not significantly affected. CBMC production of interleukins-2, -4, -5, and -10 and tumour necrosis factor-α was lower in the salmon group. There was no effect of salmon on the atopic outcomes assessed at 6 months.

Il rapido aumento dell’incidenza delle malattie cronico-degenerative è probabilmente uno dei maggiori problemi di sanità pubblica attuali. Sono numerosi gli studi che supportano il ruolo che determinati modelli alimentari ricchi in specifici alimenti o composti bioattivi hanno nella riduzione del rischio di sviluppare malattie. L’intervento di tipo nutrizionale è sempre più considerato una strategia di prevenzione primaria che risulta tra le più efficaci, convenienti e facilmente perseguibili. Recentemente, un interesse crescente si sta sempre più focalizzando su interventi dietetici personalizzati accuratamente studiati per soddisfare le specifiche esigenze nutrizionali di gruppi di popolazione target. Infatti, un approccio nutrizionale personalizzato sembra essere molto più efficace nell’ottenimento di uno specifico risultato, soprattutto in quei gruppi di popolazione target considerati più a rischio di sviluppare malattie croniche. Sulla base di queste premesse, lo scopo della presente tesi di dottorato è stato quello di valutare, attraverso approcci in vivo ed ex vivo, l'effetto del consumo di specifici alimenti e dei loro composti bioattivi nella modulazione di alcuni fattori di rischio per le malattie cronico-degenerative, in gruppi di popolazione “a rischio”. La prima parte della tesi (approccio in vivo) ha avuto l’obiettivo di valutare l’impatto di due diversi interventi dietetici in una popolazione di bambini ed adolescenti italiani affetti da iperlipidemia primaria, e quindi considerati a maggior rischio di possibili eventi cardiovascolari futuri. In primo luogo, la popolazione pediatrica in studio è stata caratterizzata per il profilo lipidico sierico e per la composizione degli acidi grassi dei fosfolipidi delle membrane eritrocitarie, mostrando delle differenze in base al sesso e al tipo specifico di iperlipidemia. Inoltre, in questi soggetti è stata osservato un basso indice omega 3 (acido eicosapentaenoico, EPA + acido docosaesaenoico, DHA: <4%), un marker che è stato suggerito essere associato a un elevato rischio di sviluppare malattie cardiovascolari. In secondo luogo, nella stessa popolazione di pazienti pediatrici iperlipidemici è stato studiato l’effetto del consumo regolare di uno specifico alimento (nocciole, HZN) o un supplemento alimentare (olio di semi di canapa, HSO), entrambi ricchi in acidi grassi insaturi e altri bioattivi, nella modulazione di diversi biomarker di rischio cardiovascolare collegati all’iperlipidemia. Entrambi gli interventi dietetici sono risultati efficaci nella gestione dell’iperlipidemia pediatrica. In particolare, abbiamo dimostrato che l’assunzione per 8 settimane di nocciole, consumate con la cuticola (HZN+S) oppure pelate (HZN-S), migliorava significativamente il profilo lipidico sierico e aveva un impatto favorevole anche sulla composizione degli acidi grassi delle membrane eritrocitarie. Inoltre, i trattamenti dietetici con le nocciole sono stati in grado di ridurre i livelli di danno al DNA, un marker di stress ossidativo. Tuttavia, le concentazioni plasmatiche di LDL ossidate (ox-LDL) non hanno subito alcun cambiamento in seguito all’intervento con le nocciole. L’effetto della supplementazione con olio di semi di canapa nella modulazione del profilo lipidico in bambini iperlipidemici è stato valutato attraverso uno studio pilota. Anche se preliminari, i risultati di questo studio hanno mostrato un miglioramento dell’indice omega 3 e della composizione degli acidi grassi delle membrane eritrocitarie in seguito all’assunzione dell’olio di semi di canapa per 8 settimane. La seconda parte della tesi di dottorato è stata svolta su modelli ex vivo presso il Dipartimento di Fisiologia e Biochimica della Nutrizione del Max Rubner-Institut (Karlsruhe, Germania). Considerando che le malattie cronico-degenerative sono spesso associate ad una risposta immunitaria compromessa, l’obiettivo di questa parte di ricerca è stato quello di studiare il potenziale effetto immunomodulatorio di diverse concentrazioni di acidi grassi polinsaturi a lunga catena (LC-PUFA), in combinazione con dosi standard di vitamina D3, in cellule immunitarie ex vivo prelevate da volontari sani. L’effetto di questi composti bioattivi è stato valutato su due meccanismi implicati nella risposta immunitaria innata, che agiscono specificatamente contro le cellule tumorali e gli agenti patogeni: l’attivita delle cellule natural killer (NK) e la fagocitosi. I risultati ottenuti hanno mostrato che gli LC-PUFA ad elevate dosi, indipendentemente dalla presenza o meno della vitamina D, interferiscono negativamente sulla capacità delle cellule NK di lisare e contrastare le cellule tumorali target, mostrando una risposta dose-dipendente. Al contrario, è stato osservato un miglioramento dell’attività fagocitaria dei monociti quando gli LC-PUFA erano coincubati con la vitamina D, suggerendo un potenziale effetto sinergico e immunomodulatorio della vitamina D. In conclusione, attraverso questa tesi di dottorato è stato possibile aumentare le conoscenze sul potenziale effetto di diete ricche in composti bioattivi nella modulazione di alcuni fattori di rischio per le malattie cronico-degenerative. Questo aspetto risulta particolarmente importante soprattutto nelle popolazioni target considerate più a rischio e per le quali un intervento nutrizionale appropriato e personalizzato viene considerato l’approccio di prevenzione primario. Studi futuri su gruppi di popolazione più ampi e volti a valutare i meccanismi d’azione specifici sono necessari al fine di fornire delle dimostrazioni aggiuntive che confermino i nostri risultati.
The rapid increase in prevalence of chronic-degenerative diseases is probably the major global health problem nowadays. Several studies have emphasized the role that dietary patterns rich in specific foods or their bioactive compounds may play in the reduction of disease risk. In fact, nutritional intervention is considered one of the most significant, easily achieved and affordable primary prevention strategy. Recently, increasing interest has been focused on personalized dietary interventions accurately designed to meet specific nutritional needs of target groups of population. The personalized approach seems to be more effective to achieve a defined outcome, especially in targeted subgroups of population considered “at higher risk” to develop chronic diseases. Based on these premises, the aim of the present Ph.D. thesis was to evaluate the effect of specific foods and their bioactives in the modulation of risk factors for chronic-degenerative diseases in “at risk” groups of population, thorough both in vivo and ex vivo approaches. The first part (in vivo approach) was focused on the impact of two different dietary interventions in a population of Italian children and adolescents affected by primary hyperlipidemia and thus considered at “higher risk” to develop cardiovascular (CV) events later in life. Firstly, the study population was characterized for serum lipid profile and fatty acid (FA) composition of red blood cell (RBC) phospholipids, showing differences according to sex and diagnosis of hyperlipidemia. Moreover, in the hyperlipidemic pediatric patients we observed an overall low omega 3 index (eicosapentaenoic acid, EPA + docosahexaenoic acid, DHA: <4%), an emerging risk factor for cardiovascular diseases (CVD). Secondly, in the same population of hyperlipidemic pediatric patients, it was investigated the effect of regular intake of a specific food (hazelnuts, HZN) or food supplement (hempseed oil, HSO) rich in unsaturated fats and other bioactives, in the modulation of different CVD biomarkers related to hyperlipidemia. We documented that both dietary treatments were effective in the management of primary hyperlipidemia at pediatric age. In particular, we showed that 8-week of hazelnuts intake, consumed with skin (HZN+S) or without skin (peeled, HZN-S), significantly improve the serum lipid profile and had a favorable impact on FAs composition of RBC membranes. In addition, HZN treatments were able to reduce the levels of DNA damage, a marker of oxidative stress, even if the concentrations of plasma oxidized LDL (ox-LDL) did not change following the HZN intervention. The effect of HSO supplementation on lipid profile of hyperlipidemic children was evaluated through a pilot study. Even if preliminary, the results from this study showed an enhancement of the omega-3 index and the RBC phospholipid composition following the intake of HSO for 8 weeks. The second part of the Ph.D thesis was performed at the Department of Physiology and Biochemistry of Nutrition of Max Rubner-Institut (Karlsruhe, Germany) and devoted to ex vivo approaches. Considering that chronic diseases are often associated to a compromised immune response, the objective of this part of research was to investigate the potential immunomodulatory properties of different concentrations of long-chain polyunsaturated fatty acids (LC-PUFA) in conjunction with standard dose of vitamin D3 on ex vivo immune cells from healthy volunteers. The effect of these bioactives was evaluated on two mechanisms crucially involved in the innate immune response that specifically act against cancer cells as well as infectious agents: the natural killer (NK) cell activity and the phagocytosis. The findings showed that high dose of LC-PUFA, independently by the presence of the vitamin D, negatively affect the lytic activity of NK cell of target cancer cells, with a dose-dependent response. On the contrary, an enhancement of phagocytic activity of monocytes was observed only when LC-PUFA were combined with vitamin D, suggesting a potential synergic and immunomodulatory role of vitamin D. In conclusion, through this Ph.D thesis it was possible to increase knowledge on the potential benefits of bioactives rich-diet in the modulation of different risk factors for chronic-degenerative diseases. This aspect is particulary important in at risk target groups of population for whom appropriate and personalized nutritional interventions are considered the primary prevention approach. Future studies on largest population groups aimed to clarify the specific mechanisms of action involved are needed to provide additional demonstration that confirm our results.

Objetivou-se avaliar o uso do grão de soja cru integral na dieta de bovinos de corte sobre o consumo e digestibilidade da matéria seca e nutrientes, fermentação ruminal, desempenho produtivo, síntese de proteína microbiana, características de carcaça qualidade da carne, perfil de ácidos graxos da carne, concentrações de parâmetros sanguíneos, excreção e composição do grão de soja presente nas fezes. Foram realizados dois experimentos, sendo que no primeiro, foram utilizados 12 bovinos Nelore, castrados, canulados no rúmen, agrupados em três quadrados latinos 4x4. No segundo experimento foram utilizados 52 bovinos Nelore, inteiros, confinados por 84 dias, em delineamento inteiramente casualizado. A dieta empregada foi a mesma, para ambos os experimentos, constituída de quatro rações, com relação volumoso/concentrado de 60/40, a qual foi composta de milho moído, farelo de soja, grão de soja cru integral, núcleo e silagem de milho. As dietas foram: G0: dieta controle sem a inclusão do grão de soja; G8, G16 e G24, com 8, 16 e 24%, respectivamente de grão de soja cru integral na ração, na matéria seca. No Experimento 1, amostras de fezes e sobras foram coletadas nos 11o, 12o 13o dia experimento, amostras de sangue e urina foram colhidas e avaliado o peso vivo. A digestibilidade foi determinada por meio de indicador interno FDAi. No Experimento 2, a cada 28 dias, os animais foram pesados e amostras de sangue foram coletadas. Os animais foram abatidos ao 85o dia e foi avaliado o peso do fígado (PFi) e peso de carcaça quente (PCQ). Vinte e quatro horas após o abate, foram mensurados o pH, e rendimento de carcaça (RC). Na desossa foi avaliada a área de olho de lombo (AOL) e a espessura de gordura subcutânea (EGS) no músculo Longissimus. Foram retiradas amostras do músculo Longissimus, para determinação do perfil de ácidos graxos na carne (PAG) extrato etéreo (EE), maciez objetiva e sensorial da carne maturada por 14 dias. Houve redução no consumo de matéria seca, matéria orgânica, de carboidratos totais e carboidratos não fibrosos nos animais alimentados com a ração G24. Foi observado efeito linear crescente no consumo de EE e efeito quadrático no consumo de proteína bruta. Houve diferença na digestibilidade aparente total do EE e carboidratos totais com a adição do grão de soja na dieta. Na fermentação ruminal, houve queda linear no valor de pH ruminal e a concentração de nitrogênio amoniacal ruminal foi maior para os animais que receberam a dieta controle em relação às dietas com grão de soja, havendo um maior valor de ácidos graxos de cadeia curta para o grupo controle. O teor de colesterol sanguíneo aumentou linearmente com a inclusão do grão e no primeiro experimento também houve aumento do colesterol HDL. Para desempenho, atributos de carcaça e medidas de ultrassonografia não foi observado efeito de acordo com a dieta. Houve efeito quadrático para força de cisalhamento (FC), com maior valor para a carne dos animais que receberam 8% do grão de soja, Houve uma discreta melhora no perfil lipídico para animais recebendo o grão e na analise sensorial a inclusão do grão ao nível de 24% a MS da dieta melhorou a textura das carnes. A utilização de diferentes níveis de grão de soja nas rações alterou o consumo, digestibilidade, metabolismo e características de qualidade da carne.
The objective of this study was to evaluate the use of whole raw soybean grain in the diet of beef cattle on consumption and digestibility of dry matter and nutrients, ruminal fermentation, growth performance, microbial protein synthesis, carcass traits, meat quality, profile of fatty acids in meat, concentrations of blood parameters, excretion and composition of soybean in the faeces. Two experiments were conducted and in the first one were used 12 Nellore, castrated, cannulated in the rumen, grouped in three 4x4 Latin squares. In the second experiment were used 52 Nellore, not castrated, confined for 84 days in a completely randomized delineation. The diet used was the same for both experiments, consisting of four rations, with the forage / concentrate ratio of 60/40, which was composed of ground corn, soybean meal, whole raw soybean grain, nucleos and corn silage. The diets were: G0: control diet without the inclusion of soybean, G8, G16 and G24, with 8, 16 and 24% respectively of whole raw soybean grain in ration, in the dry matter. In the first experiment, fecal samples and remains were collected on the 11th, 12th and 13th day of experiment, blood and urine samples were collected and the body weight was analyzed. The digestibility was determined by an internal indicator iADF. In Experiment 2, every 28 days, the animals were weighed and blood samples were collected. The animals were slaughtered at 85 days and were weighed the liver (PFi) and hot carcass weight (HCW). Twenty-four hours after the slaughter, the pH was measured, likewise the carcass yield (CY). Deboning was evaluated in rib eye area (REA) and subcutaneous fat thickness (SFT) in the Longissimus muscle. Samples were taken from the Longissimus for determining the fatty acid profile in meat (PAG) in ether extract (EE), objective tenderness and sensorial in aged beef for 14 days. There was a reduction in the dry matter intake, organic matter, total carbohydrates and non-fibrous carbohydrates in animals fed with feed with ration G24. It was observed an increasing linear effect on the EE intake and a quadratic effect on crude protein. Also difference in the apparent digestibility of total carbohydrates and total EE. On ruminal fermentation, there was a linear decline in ruminal pH and ruminal ammonia concentration was higher for animals that received the control diet compared to diets with soybean, with a value greater short-chain fatty acids for the group control. The level of blood cholesterol increased linearly with the addition of grain and in the first experiment there was also increase of HDL cholesterol. For performance, attributes of carcass and measures of ultrasound effect were not observed according to the diet. There was a quadratic effect for shear force (FC), with higher value for the meat of animals that received 8% of the soybean. There was a slight improvement in lipid profile for animals receiving the grain and the inclusion in the sensory analysis of the grain at 24% of diet DM improved the texture of the meat. The use of different levels of whole raw soybean grain in the beef cattle diet changed the intake, digestibility, metabolism and meat quality characteristics.

BACKGROUND AND AIM: Diet, in particular dietary fats, and cardiovascular disease (CVD) are closely related. Dietary fats might be captured by measuring blood fatty acid profiles. The role of diet as well as the role of blood fatty acid (FA) levels, in CVD aetiology is still uncertain. Aims of this thesis were to investigate in a large cohort of 60-year-old Swedish men and women: 1) The association between self-reported dietary intake, with a specific focus on foods rich in fat, and selected serum cholesterol ester FAs (Project I); 2) The relation between self-reported intake of specific types of dietary fats (primary aim) and fruit and vegetables (secondary aim) and incident of CVD and all-cause mortality (Project II); 3) The relation between serum cholesterol FAs, with a specific focus on polyunsaturated FAs (PUFA)eicosapentaenoic acid (EPA), docosaesaenoic acid (DHA), linoleic (LA) and linolenic (ALA) acid and incident CVD and all-cause mortality (Project III). METHODS: Data collected between 1997 and 1998 from 4,232 individuals (2,039 men and 2,139 women) aged 60, randomly selected from Stockholm County were used. The participants were followed regarding incident CVD up to 31st December 2012 using national registers yielding 359 incident CVD cases and 595 deaths. From nutritional data, collected by questionnaires, we created: 1) five diet scores reflecting intake of saturated fats in general, and fats from dairy, fish, processed meat and vegetable oils and margarines (Project I, II) 2) binary variables classifying study participants into exposed and unexposed and evaluating 16 specific dietary factors (Project II). Gas chromatography was used to assess 13 FAs in serum cholesterol esters (Project I, III). Association between each diet score and specific FAs was assessed by percentile differences (PD) with 95% confidence intervals (CI) at the 10th, 25th, 50th, 75th, and 90th percentile of each FA across levels of diet scores using quantile regression (Project I). Crude and adjusted Cox proportional hazards models were used to estimate hazard ratios (HR) with 95% CI in the association between specific self-reported dietary fats (diet scores and single dietary items), fruit and vegetables intake (Project II) and serum PUFA (Project III) and incidence of CVD and all-causes mortality. RESULTS: In men and women combined, fish intake was associated with high serum proportions of EPA (50thPD=31.41, 95% CI= 27.77; 35.05) and DHA (50thPD=10∙51, 95% CI= 9.40; 11.62). Vegetable fat intake was associated with high serum proportion of total PUFA (50thPD 36.34, 95% CI= 22.77;49.92) and low proportion of total SFA (50thPD=11.33, 95%CI= 14.92;7.73). (Project I) In women, an increased risk of CVD was related to high consumption of spread butter or margarine (≥10g/day vs <10g/day), HR=1.49, CI=1.02 ; 2.20, and oily potatoes (≥2 times/week vs <2time/week), HR=2.00, CI=1.11;3.60. In men, an increased risk of early death was related to the consumption of butter (vs margarine), HR=1.28, CI=1.01; 1.62, high consumption of spread butter or margarine, HR=1.57, CI=1.23; 2.02 and egg consumption ≥4 times/week (vs <4times/week), HR=1.53, CI=1.15;2.02. In men, daily intake of fruits (vs <1time/day) was inversely related to early death, HR=0.75, CI=0.60; 0.94. (Project II) High serum EPA and DHA proportions were inversely associated with CVD in women (for EPA HR= 0.79, 95% CI 0.64; 0.97; for DHA HR= 0.74 0.61; 0.89) but not in men. Inverse associations with early death were also noted in men for high serum EPA proportion, HR=0.82, 95% CI 0.71;0.95; and DHA proportion, HR= 0.82, 95%CI= 0.71;0.94, and in women for high serum EPA proportion, HR=0.79, 95%CI= 0.65;0.96, and DHA proportion, HR= 0.78, 95% CI= 0.66;0.93. High serum ALA proprotion was associated with moderately increased of CVD incidence, HR= 1.16, 95% CI=1.02;1.32 in women whereas high serum LA proportion was associated with reduced all-cause mortality in men, HR= 0.73 95% CI=0.64;0.83. (Project III). CONCLUSION: Based on our results, self-reported intake of fish and vegetable fats was clearly associated with serum PUFA. High intake of specific foods and not fats in general may have negative effects on CVD for women and all-causes mortality for men, whereas fruit may reduce mortality only in men. Similarly serum EPA, DHA and LA were protective for CVD and all-causes mortality with gender difference whereas serum ALA might be associated with increased of CVD in only women.

Soybean oil (SBO), menhaden fish oil (FO), chicken fat (CF) or hydrogenated soybean oil (HSBO) were incorporated at 5% of the total diet to study changes in the immunological status of both Japanese quail (JAP) and Bobwhite quail (BOB). The SBO diet, in which 66% of the total fatty acids were polyunsaturated fatty acids (PUFA), was rich in linoleic acid (LA 18:2 n-6), alpha-linolenic acid (ALA 18:3 n-3) and low in saturated fatty acid (SFA). The FO diet which contained about 50% PUFA, had only 40% n-6 fatty acids and 8% n-3 PUFA. The trans fatty acid isomers and other monounsaturated fatty acids (MUFA) were high in the HSBO diet. The diet containing CF provided a relatively balanced fatty acid composition with 18% SFA, 31% MUFA and 50% PUFA. Plasma fatty acid and hepatic fatty acid profiles consistently reflected their respective dietary lipid treatments. There were no differences in the fatty acid profile between blood and liver within respective dietary treatments in the two species. Dietary fatty acids had no effect on antibody titers against sheep red blood cells (SRBC) at 1, 2 and 8 months following the start of dietary lipid treatment in JAP. However, female JAP fed FO had a significantly (p< 0.05) higher antibody production compared to the other dietary lipid treatments at 4 months following the start of fatty acids supplementation. BOB fed either FO or SBO diets had a higher immunoglobulin G production compared to birds fed the CF diet. The total antibody titer was significantly higher in BOB fed SBO compared to CF. Dietary fatty acids had a significant effect on cell-mediated immunity (CMI) as accessed by toe web thickness 24 hours post intradermal injection of phytohemagglutinin-P (PHA) in both JAP and BOB. In general, birds fed a FO diet had a significantly higher CMI response than those fed HSBO. A diet high in n-3 PUFA increased the index of cutaneous basophil hypersensitivity (CBH), while the high trans fatty acid isomers suppressed the CBH response. By observing a CBH response over a 72-hour period in JAP, it was concluded that quail fed CF or SBO had a different peak response time (12 hours post PHA challenge) and amplitude compared with those fed FO or HSBO (24 hours post PHA challenge). Phagocytic ability was not affected by dietary lipid treatments in BOB while the quail fed FO diet had a faster carbon clearance rate. The FO fed JAP had a significantly higher response (p< 0.05) to concanavalin A ensiformis (CONA) compared to HSBO fed birds. There was no difference in B lymphocyte proliferation stimulated by lipopolysacchride (LPS) in female JAP, whereas it was significantly higher in male JAP fed SBO compared to those fed FO and HSBO. Phorbol 12-myristate 13-acetate/ionomycin calcium salt (PMA/ION) was used to nonspecifically stimulate cell proliferation by increasing chromosome mitosis. Dietary FO or HSBO suppressed cell proliferation stimulated by PMA/ION. However, JAP fed SBO or CF had a significantly higher PMA/ION stimulated lymphocyte proliferation compared those fed FO or HSBO. In male BOB, the FO fed birds had the highest response to all mitogens. In contrast, female BOB did not show any dietary effects by lymphocyte proliferation. Consistent with JAP, BOB fed HSBO had depressed lymphocytes proliferation in response to various mitogens stimulation. In general, female birds had a higher plasma total protein (PTP) and lower pack cell volume (PCV) compared to their males counterparts in both BOB and JAP. In summary, in in vivo experiments, feeding a diet high in menhaden fish oil that is rich in n-3 PUFA enhanced the CMI. There was a minimal effect on antibody production caused by feeding n-3 PUFA in JAP since a significant treatment effect was only found at one sampling period, while BOB were more sensitive to dietary lipid manipulation and had a higher antibody production with SBO or FO treatments. Dietary lipids exerted different effects in the two species in in vitro experiments. While both BOB and JAP fed FO had higher lymphocyte proliferation to CON A mitogen compared to those fed HSBO, only male BOB showed a higher proliferation to LPS. Feeding HSBO that contained a higher content of trans fatty acid isomers, MUFA, but lower PUFA content resulted in the lowest lymphocyte proliferation to various mitogens in both BOB and JAP.
Ph. D.

Hypertension is a widespread condition which may cause cardiovascular events when left untreated. If high blood pressure (BP) is noticed at all, it is mostly only sub-optimally controlled making nutritional interventions a cost-effective and safe preventive measure and an alternative to medical treatment. Previous studies have shown that increased fibre consumption reduces BP which was particularly effective in hypertensive subjects. Fibres are indigestible and hence are available for fermentation by the colonic microbiota which produces the short chain fatty acids (SCFAs) acetate, propionate and butyrate. Intriguingly, recent studies carried out in mice showed that SCFAs can reduce BP. Therefore, we hypothesised that gut microbiota-derived SCFAs can (1) reduce BP in middle-aged male volunteers and (2) influence the protein expression of BP regulatory systems in a cellular model. As the development of a cellular angiotensin II-induced hypertension model was unsuccessful, the effects of SCFAs on a molecular level were assessed in unstimulated human aortic endothelial cells (HAECs). The expression of proteins involved in the BP regulating renin angiotensin system (RAS) was assessed by western blotting. Additionally, a human supplementation trial is being carried out looking at the acute consumption of a low (0.16 g) and high (2.35 g) propionate dose on BP and other cardiovascular markers in middle-aged male volunteers. In vitro work showed that SCFAs did not affect RAS expression in HAECs. However, acute propionate supplementation influenced BP and its regulation. Preliminary data show, that while a high propionate dose led to increases in plasma propionate by on average 4 µM and acetate levels with concurrent increases in BP, arterial stiffness and plasma renin concentration, a low propionate dose resulted in plasma propionate increases of about 0.5 µM with simultaneous reductions in systolic BP. Taken together, these results suggest that SCFAs play a regulatory role in the homoeostasis of BP.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro
Introdução: o óxido nítrico (NO) é um gás inorgânico com uma meia-vida curta e tem um papel crítico na manutenção da homeostase vascular e fluidez sanguínea. O NO é sintetizado a partir do aminoácido L-arginina por uma família de enzimas NO sintases (NOS). Estudos têm mostrado que eritrócitos expressam NOS endotelial (eNOS) funcional, que serve como uma fonte de NO intraluminal. Além disso, eritrócitos participam da defesa antioxidante removendo os radicais livres e prevenindo o dano oxidativo às membranas biológicas e a destruição do NO. Dietas hiperlípidicas estão associadas a um risco aumentado de doença cardiovacular e síndrome metabólica, mas os exatos mecanismos não estão completamente esclarecidos. O objetivo deste estudo foi investigar os efeitos de diferentes dietas hiperlípidicas na via L-arginina-NO e o estresse oxidativo em eritrócitos de camundongos. Metodologia: camundongos machos C57BL/6 de três meses de idade receberam diferentes dietas por 10 semanas: dieta normolipídica ou dieta hiperlipídica contendo banha de porco (HB), óleo de oliva (HO), óleo de girassol (HG) ou óleo de canola (HC). Foram analisados o transporte de L-arginina mediado pelos transportadores catiônicos y+ e y+L, a atividade da NOS, a expressão da eNOS e da NOS induzível (iNOS), a formação de substâncias reativas ao ácido tiobarbitúrico (TBARS) e a atividade das enzimas antioxidantes catalase (CAT) e superóxido dismutase (SOD). Resultados: o transporte total de L-arginina estava aumentado no grupo HO em comparação aos controles e aos outros grupos com dieta hiperlipídica. Quando o transporte foi fracionado, o sistema y+ estava mais ativado no grupo HO em relação aos controles e outros grupos que receberam dieta hiperlipídica. O transporte de L-arginina via sistema y+L estava maior nos grupos HO, HG e HC comparados aos grupos controle e HB. Adicionalmente, a atividade basal da NOS e a expressão de eNOS estavam aumentadas em eritrócitos independente do tipo de dieta hiperlípidica insaturada. Observou-se uma maior expressão da iNOS no grupo HO comparado ao controle. Em contraste, o grupo HB apresentou uma inibição da via L-arginina-NO. A análise da peroxidação lipídica, através da formação de TBARS, e da atividade da enzima antioxidante CAT não revelou diferenças entre os grupos, ao contrário do grupo HO, que induziu uma ativação de outra enzima antioxidante, a SOD. Conclusões: o presente estudo proporciona a primeira evidência de que os sistemas y+ e y+L regulam o transporte aumentado de L-arginina em eritrócitos de camundongos do grupo HO. Além disso, todas as dietas hiperlipídicas insaturadas induzem um aumento da atividade basal da NOS associada a uma expressão elevada da eNOS. É possível que diferentes mudanças na composição lipídica da membrana plasmática induzidas pelas dietas possam afetar transportadores e enzimas nos eritrócitos. Além disso, a inibição da via L-arginina-NO no grupo HB pode contribuir para o desenvolvimento da aterosclerose, enquanto dietas hiperlipídicas insaturadas podem ter um efeito protetor via aumento da geração de NO.
Introduction: nitric oxide (NO) is an inorganic gas with a short half life that plays a critical role in maintaining vascular homeostasis and blood fluidity in physiological conditions. NO is synthesized from the cationic amino acid L-arginine by a family of enzymes: nitric oxide synthase (NOS). Studies have shown that red blood cells (RBCs) express functional endothelial NOS (eNOS), which potentially serves as an intraluminal NO source. Moreover, circulating RBCs participate in antioxidant defence, scavenging oxygen free radicals and preventing oxidative damage to biological membranes and NO destruction. High fat diets are associated with an increased risk of cardiovascular disease and metabolic syndrome, but the exact mechanisms are not completely clear. The objective of this study was to investigate the effects of different high fat (HF) diets in the RBC L-arginine-NO pathway and in oxidative stress in C57BL/6 mice. Methods: three-month-old male C57BL/6 mice were fed different diets for a 10-week period: a standard diet or high-fat (HF) diet containing lard oil (HF-L), olive oil (HF-O), sunflower oil (HF-S) or canola oil (HF-C). Studies of L-arginine transport, mediated by cationic transport systems y+ and y+L, basal activity of NOS, expression of eNOS and inducible NOS (iNOS), thiobarbituric acid reactive substances (TBARS) formation, and antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD) activities in RBCs were analysed in these groups. Results: total L-arginine influx into RBCs was upregulated in the HF-O group compared to controls and other HF diet groups. When transport systems were fractionated, there was a higher activation of system y+ in the HF-O group in relation to controls and other HF diet groups. L-arginine transport via system y+L in RBCs was increased in the HF-O, HF-S and HF-C groups compared to controls and the HF-L group. In addition, NOS activity and eNOS expression were enhanced in RBCs, independent of unsaturated HF diets. An overexpression of iNOS was observed in the HF-O group compared with controls. In contrast, the HF-L group showed an inhibition of the RBC L-arginine-NO pathway. The analysis of lipid peroxidation and antioxidant enzyme catalase activity revealed no differences among the groups studied. On the other hand, HF-O induced activation of another antioxidant enzyme, superoxide dismutase (SOD). Conclusions: this study provides the first evidence that systems y+ and y+L mediate increased L-arginine transport into mice RBCs from the HF-O group. Moreover, all unsaturated high-fat diets can induce an increase in basal NOS activity associated with an overexpression of eNOS. It is possible that changes in the lipid composition of the plasmatic membrane induced differently by HF diets could affect transporters and enzymes in RBCs. An inhibition of the L-arginine-NO pathway in HF-L group could contribute to the development of atherosclerosis, while HF unsaturated diets may have a protector effect via enhanced generation of NO.

Cardiovascular disease rates (CVD) are high in the UK, particularly in men. National surveys report a dietary imbalance of fats and antioxidant micronutrients, which may adversely influence blood fatty acid profile and risk for CVD. We investigated relationships between saturated and unsaturated dietary fat, and antioxidant (retinol, alpha-tocopherol, beta-carotene, iron, zinc, selenium, magnesium and copper) intakes, and blood glucose, lipoproteins and cell membrane fatty acids in a homogenous group of educated, healthy Caucasian men and women. Volunteers (20 to 50 years, men 52, women 52) were recruited from staff and students of London Metropolitan University. Following correction for under-reporting 40 women and 31 men were included in the dietary analysis. Dietary intake, body composition, blood pressure, fasting blood glucose and lipids, and fatty acid composition of plasma and blood cell membranes (red blood cells, and platelets) were analysed. The men consumed more saturated fat (11.0 vs 9.8 %energy intake, p<0.05), and exhibited elevated glucose (5.41 vs 4.81 mmol/I, p