Fiona Lewis

Effect of low doses of long-chain n−3 PUFAs on endothelial function and arterial stiffness: a randomized controlled trial

BACKGROUND The dietary intake of n-3 (omega-3) long-chain PUFAs (LC-PUFAs) from fish may improve endothelial function and arterial stiffness. OBJECTIVE The objective was to test the hypothesis that increasing intakes of n-3 LC-PUFAs-equivalent to the consumption of 1, 2, or 4 portions of oily fish per week-favorably affects endothelial function and arterial stiffness. DESIGN A parallel-design, randomized, double-blind study compared daily doses of 0.45, 0.9, and 1.8 g n-3 LC-PUFAs (EPA:DHA ratio of 1.51:1) with placebo (refined olive oil). The primary and secondary outcomes were changes in flow-mediated dilatation (FMD) of the brachial artery, arterial stiffness, and blood pressure. Nonsmoking men (n = 142) and women (n = 225) aged 45-70 y were randomly assigned to treatment for 12 mo; 312 subjects completed the intervention. RESULTS Compliance with the intervention was corroborated by significant dose-dependent increases in the proportions of EPA and DHA in erythrocyte lipids and a 16.5% reduction in serum triacylglycerol concentrations with 1.8 g n-3 LC-PUFAs/d. FMD was lower in men than in women (P < 0.0001) and decreased with age (ρ = 0.270, P < 0.001) but was not significantly (P = 0.781) related to n-3 LC-PUFA intake. The mean changes in FMD (95% CIs) compared with placebo were 0.1% (-0.9%, 1.1%), -0.3% (-1.3%, 0.6%), and -0.3% (-1.3%, 0.7%) with daily intakes of 0.45, 0.9, and 1.8 g n-3 LC-PUFAs, respectively. No significant treatment effects were noted for arterial stiffness and central mean or 24-h ambulatory blood pressure. CONCLUSION Intakes of n-3 LC-PUFAs ≤1.8 g/d do not improve endothelial function in healthy adults. The trial is registered at controlled-trials.com as ISRCTN66664610.

Genetic variation at the FADS1-FADS2 gene locus influences delta-5 desaturase activity and LC-PUFA proportions after fish oil supplement

Delta-5 and delta-6 desaturases (D5D and D6D) are key enzymes in endogenous synthesis of long-chain PUFAs. In this sample of healthy subjects (n = 310), genotypes of single nucleotide polymorphisms (SNPs) rs174537, rs174561, and rs3834458 in the FADS1-FADS2 gene cluster were strongly associated with proportions of LC-PUFAs and desaturase activities estimated in plasma and erythrocytes. Minor allele carriage associated with decreased activities of D5D (FADS1) (5.84 × 10−19 ≤ P ≤ 4.5 × 10−18) and D6D (FADS2) (6.05 × 10−8 ≤ P ≤ 4.20 × 10−7) was accompanied by increased substrate and decreased product proportions (0.05 ≤ P ≤ 2.49 × 10−16). The significance of haplotype association with D5D activity (P = 2.19 × 10−17) was comparable to that of single SNPs, but haplotype association with D6D activity (P = 3.39 × 10−28) was much stronger. In a randomized controlled dietary intervention, increasing eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) intake significantly increased D5D (P = 4.0 × 10−9) and decreased D6D activity (P = 9.16 × 10−6) after doses of 0.45, 0.9, and 1.8 g/day for six months. Interaction of rs174537 genotype with treatment was a determinant of D5D activity estimated in plasma (P = 0.05). In conclusion, different sites at the FADS1-FADS2 locus appear to influence D5D and D6D activity, and rs174537 genotype interacts with dietary EPA+DHA to modulate D5D.

SFAs do not impair endothelial function and arterial stiffness

Background: It is uncertain whether saturated fatty acids (SFAs) impair endothelial function and contribute to arterial stiffening. Objective: We tested the effects of replacing SFAs with monounsaturated fatty acids (MUFAs) or carbohydrates on endothelial function and arterial stiffness. Design: With the use of a parallel-designed randomized controlled trial in 121 insulin-resistant men and women, we measured vascular function after 1 mo of consumption of a high-SFA (HS) diet and after 24 wk after random assignment to the HS diet or diets that contained <10% SFAs and were high in either MUFAs or carbohydrates. The primary outcome was a change in flow-mediated dilation (FMD), and secondary outcomes were changes in carotid to femoral pulse wave velocity (PWV) and plasma 8-isoprostane F2α-III concentrations. Results: For 112 participants with data available for analysis on the specified outcomes, no significant differences were shown. FMD with the HS reference diet was 6.7 ± 2.2%, and changes (95% CIs) after 6 mo of intervention were +0.3 (−0.4, 1.1), −0.2 (−0.8, 0.5), and −0.1 (−0.6, 0.7) with HS, high-MUFA (HM), and high-carbohydrate (HC) diets, respectively. After consumption of the HS reference diet, the geometric mean (±SD) PWV was 7.67 ± 1.62 m/s, and mean percentages of changes (95% CIs) were −1.0 (−6.2, 4.3) with the HS diet, 2.7 (−1.4, 6.9) with the HM diet, and −1.0 (−5.5, 3.4) with the HC diet. With the HS reference diet, the geometric mean (±SD) plasma 8-isoprostane F2α-III concentration was 176 ± 85 pmol/L, and mean percentage of changes (95% CIs) were 1 (−12, 14) with the HS diet, 6 (−5, 16) with the HM diet, and 4 (−7, 16) with the HC diet. Conclusion: The replacement of SFAs with MUFAs or carbohydrates in healthy subjects does not affect vascular function. This trial was registered at Current Controlled Trials (http://www.controlled-trials.com/ISRCTN) as ISRCTN 29111298.

Successful Manipulation of the Quality and Quantity of Fat and Carbohydrate Consumed by Free-Living Individuals Using a Food Exchange Model

Our objective in this study was to develop and implement an effective intervention strategy to manipulate the amount and composition of dietary fat and carbohydrate (CHO) in free-living individuals in the RISCK study. The study was a randomized, controlled dietary intervention study that was conducted in 720 participants identified as higher risk for or with metabolic syndrome. All followed a 4-wk run-in reference diet [high saturated fatty acids (SF)/high glycemic index (GI)]. Volunteers were randomized to continue this diet for a further 24 wk or to 1 of 4 isoenergetic prescriptions [high monounsaturated fatty acids (MUFA)/high GI; high MUFA/low GI; low fat (LF)/high GI; and LF/low GI]. We developed a food exchange model to implement each diet. Dietary records and plasma phospholipid fatty acids were used to assess the effectiveness of the intervention strategy. Reported fat intake from the LF diets was significantly reduced to 28% of energy (%E) compared with 38%E from the HM and LF diets. SF intake was successfully decreased in the HM and LF diets to < or =10%E compared with 17%E in the reference diet (P = 0.001). Dietary MUFA in the HM diets was approximately 17%E, significantly higher than in the reference (12%E) and LF diets (10%E) (P = 0.001). Changes in plasma phospholipid fatty acids provided further evidence for the successful manipulation of fat intake. The GI of the HGI and LGI arms differed by approximately 9 points (P = 0.001). The food exchange model provided an effective dietary strategy for the design and implementation across multiple sites of 5 experimental diets with specific targets for the proportion of fat and CHO.

Adiponectin Gene Variant Interacts with Fish Oil Supplementation to Influence Serum Adiponectin in Older Individuals

Marine n3 polyunsaturated fatty acids (PUFAs) activate the transcription factor peroxisome proliferator-activated receptor (PPARγ), which modulates the expression of adiponectin. We investigated the interaction of dietary n3 PUFAs with adiponectin gene (ADIPOQ) single nucleotide polymorphism (SNP) genotypes as a determinant of serum adiponectin concentration. The Modulation of Atherosclerosis Risk by Increasing Doses of n3 Fatty Acids study is a parallel design, double-blind, controlled trial. Serum adiponectin was measured in 142 healthy men and 225 women aged 45-70 y randomized to treatment with doses of 0.45, 0.9, and 1.8 g/d 20:5n3 and 22:6n3 (1.51:1), or placebo for 12 mo. The 310 participants who completed the study were genotyped for 5 SNPs at the ADIPOQ locus: -11391 G/A (rs17300539), -11377 C/G (rs266729), -10066 G/A (rs182052), +45 T/G (rs2241766), and +276 G/T (rs1501299). The -11391 A-allele was associated with a higher serum adiponectin concentration at baseline (n = 290; P < 0.001). The interaction between treatment and age as a determinant of adiponectin was significant in participants aged >58 y after the highest dose (n = 92; P = 0.020). The interaction between +45 T/G and treatment and age was a nominally significant determinant of serum adiponectin after adjustment for BMI, gender, and ethnicity (P = 0.029). Individuals homozygous for the +45 T-allele aged >58 y had a 22% increase in serum adiponectin concentration compared with baseline after the highest dose (P-treatment effect = 0.008). If substantiated in a larger sample, a diet high in n3 PUFAs may be recommended for older individuals, especially those of the +45 TT genotype who have reported increased risk of hypoadiponectinemia, type 2 diabetes, and obesity.

Interaction between a CSK Gene Variant and Fish Oil Intake Influences Blood Pressure in Healthy Adults

Blood pressure is a heritable determinant of cardiovascular disease (CVD) risk. Recent genome-wide association studies have identified several single-nucleotide polymorphisms (SNPs) associated with blood pressure, including rs1378942 in the c-Src tyrosine kinase (CSK) gene. Fish oil supplementation provides inconsistent protection from CVD, which may reflect genetic variation. We investigated the effect of rs1378942 genotype interaction with fish oil dosage on blood pressure measurements in the MARINA (Modulation of Atherosclerosis Risk by Increasing doses of N-3 fatty Acids) study, a parallel, double-blind, controlled trial in 367 participants randomly assigned to receive treatment with 0.45, 0.9, and 1.8 g/d eicosapentaenoic acid [EPA (20:5n-3)] and docosahexaenoic acid [DHA (22:6n-3)] (1.51:1) or an olive oil placebo for 12 mo. A total of 310 participants were genotyped. There were no significant associations with blood pressure measures at baseline; however, the interaction between genotype and treatment was a significant determinant of systolic blood pressure (SBP) (P = 0.010), diastolic blood pressure (DBP) (P = 0.037), and mean arterial blood pressure (MABP) (P = 0.014). After the 1.8 g/d dose, noncarriers of the rs1378942 variant allele showed significantly lower SBP (P = 0.010), DBP (P = 0.016), and MABP (P = 0.032) at follow-up, adjusted for baseline values, than did carriers. We found no evidence of SNP genotype association with endothelial function (brachial artery diameter and flow-mediated dilatation), arterial stiffness (carotid-femoral pulse wave velocity and digital volume pulse), and resting heart rate. A high intake of EPA and DHA could help protect noncarriers but not carriers of the risk allele. Dietary recommendations to reduce blood pressure in the general population may not necessarily benefit those most at risk. This trial was registered at controlled-trials.com as ISRCTN66664610.

Impact of the amount and type of fat and carbohydrate on vascular function in the RISCK study

Change - 0.2 - 1.4, 0.9 0.3 - 0.8, 1.4 - 0.3 - 1.7, 1.1 PWV (m/s) (n 27) (n 42) (n 36) 0.6 - 0.2, 1.3 - 0.1 - 0.7, 0.5 - 0.6 * - 1.2, 0.1 Value was significantly different from that for the SFA treatment (Bonferronis multiple comparison test): *P < 0.05.

A persistent vulval ulcer

An 84 year old woman presented with a seven month history of a gradually enlarging, tender, shallow ulcer on the outer aspect of the right labium majus (fig 1⇓). She reported no other active or previous skin or mucosal problems and had a background of ischaemic heart disease, hypertension, hypercholesterolaemia, and type II diabetes. She was taking lisinopril, indapamide, atenolol, atorvastatin, aspirin, isosorbide mononitrate, nicorandil, metformin, and omeprazole. She was a non-smoker and there was no family history of skin disease. ### 1. What should you cover in the history and examination of a patient with vulval ulceration? #### Short answer Take a detailed history. A thorough examination of the vulva, vagina, and perianal area is needed, followed by a general skin examination, including other mucosal sites. #### Discussion Ask about the onset, triggers, and duration of the ulcer as well as the presence of fever or systemic symptoms. Has the patient had a previous episode? Does the patient have any other immune mediated problems? What personal care products does the patient use? Have smear test results been normal? Does the patient have a new partner? A full drug, sexual, and travel history could give important clues. Examine the vulva, noting the number and size of ulcers. Is there associated excoriation, fissuring, or oedema? In addition, examine the vagina, perineum, perianal skin, and inguinal lymph nodes. A general examination of the skin, eyes, and mouth may also provide vital clues to the diagnosis. For example, eczema at other sites may raise the possibility of contact dermatitis. ### 2. What differential diagnoses should you consider in a patient with vulval ulceration? #### Short answer Differential diagnoses include infection (such as herpes simplex), inflammation, cancer, trauma, and iatrogenic (drug induced) causes. Infection is a more …

Impact of the amount and type of fat and carbohydrate on serum lipids in the RISCK study

RISCK is a multi-centre randomised controlled dietary intervention in subjects at increased risk of metabolic syndrome that compares the effects of diets with a reduced SFA content that vary according to total fat, MUFA content and glycaemia index (GI). The results are reported for changes in serum lipids among the participants who completed the 6-month dietary intervention Baseline serum lipid measurements were undertaken for all subjects, subsequently all subjects followed a high-SFA (HS) diet for 1 month, after which period a further set of lipid measurements were completed. The mean serum cholesterol concentration (mmol/l) was 5.59 (SD 0.94) at screening and 5.61 (SD 0.97) at the end of the run-in period. The subjects were then randomized to one of five dietary interventions for a 6-month period: an HS–high-GI (HGI) similar to the run-in diet or one of four isoenergetic interventions (high-MUFA (HM)–HGI; HM–low-GI (LGI); low-fat (LF)–HGI; LF–LGI). The target intake for total fat was 38% energy (%E) in the HS and HM diets and 28%E in the LF diets. The HM and LF diets were designed to reduce dietary SFA to 10%E with a planned MUFA intake of 18%E in the HM diets. The target GI differential between the HGI and LGI groups was approximately 12 points. Serum lipid concentrations were determined in a single laboratory following run-in and at the end of the intervention.

APOE4 Genotype Exerts Greater Benefit in Lowering Plasma Cholesterol and Apolipoprotein B than Wild Type (E3/E3), after Replacement of Dietary Saturated Fats with Low Glycaemic Index Carbohydrates

We examined the impact of APOE genotype on plasma lipids and glucose in a secondary analysis of data from a five-arm, randomised controlled, parallel dietary intervention trial (‘RISCK’ study), to investigate the impact of replacing saturated fatty acids (SFA) with either monounsaturated fat (MUFA) or carbohydrate of high or low glycaemic index (GI) on CVD risk factors and insulin sensitivity. We tested the impact of APOE genotype (carriage of E2 and E4 alleles versus E3/E3), determined retrospectively, on plasma lipids, lipoproteins and glucose homeostasis at baseline (n = 469), and on the change in these variables after 24 weeks of dietary intervention (n = 389). At baseline, carriers of E2 (n = 70), E4 (n = 125) and E3/E3 (n = 274) expressed marked differences in total plasma cholesterol (TC, p = 0.001), low density lipoprotein cholesterol (LDL-C, p < 0.0001), apolipoprotein B (apo B, p < 0.0001) and total to high density lipoprotein cholesterol ratio (TC:HDL-C, p = 0.002), with plasma concentrations decreasing in the order E4 > E3/E3 > E2. Following intervention, there was evidence of a significant diet x genotype interaction with significantly greater decreases in TC (p = 0.02) and apo B (p = 0.006) among carriers of E4 when SFA was replaced with low GI carbohydrate on a lower fat diet (TC −0.28 mmol/L p = 0.03; apo B −0.1 g/L p = 0.02), and a relative increase in TC (in comparison to E3/E3) when SFA was replaced with MUFA and high GI carbohydrates (TC 0.3 mmol/L, p = 0.03). Among carriers of E2 (compared with E3/E3) there was an increase in triacylglycerol (TAG) when SFA was replaced with MUFA and low GI carbohydrates 0.46 mmol/L p = 0.001). There were no significant interactions between APOE genotype and diet for changes in indices of glucose homeostasis. In conclusion, variations in APOE genotype led to differential effects on the lipid response to the replacement of SFA with MUFA and low GI carbohydrates.