Study protocol for a 15-week randomised controlled trial assessing the independent effects of high-cholesterol and high-saturated fat diets on LDL cholesterol.

INTRODUCTION: Previous research has associated high dietary cholesterol intake with raised low-density lipoprotein cholesterol (LDL-C) and thus increased risk for cardiovascular disease (CVD). Emerging research suggests that it is saturated fat, not dietary cholesterol, associated with increased CVD risk. Despite being high in cholesterol, eggs, low in saturated fat, are not adversely associated with blood lipids or CVD risk. This paper describes a randomised controlled counter-balanced, cross-over trial assessing the effects of a high-cholesterol/low-saturated fat (egg) diet and a low-cholesterol/high-saturated fat diet (egg free) on blood lipids and lipoproteins, while accounting for physical activity levels which can also influence these parameters. The primary aim is to demonstrate that high cholesterol intake (from eggs) within a healthy, low-saturated fat diet does not adversely affect blood lipid levels and lipoprotein profiles. Instead, we propose that adverse effects on these parameters are mediated by saturated fat intake. The secondary aim is to explore relationships between changes in blood lutein and zeaxanthin concentrations and alterations in physical activity, examining whether changes in physical activity mediate effects on blood lipids and lipoproteins. METHODS AND ANALYSIS: Fifty-two adults aged 18-60 years with LDL-C less than 3.5 mmol/L will be randomly allocated to three isocaloric diets for 5 weeks each: a high-cholesterol (600 mg)/low-saturated fat (6%) (egg) diet, a low-cholesterol (300 mg)/high-saturated fat (12%) (egg free) diet and a control diet that is high in both cholesterol (600 mg) and saturated fat (12%). Lipid and lipoprotein levels, lipoprotein size and concentrations, blood pressure, blood glucose, physical activity levels, and plasma lutein and zeaxanthin concentrations will be measured. Treatment effects will be analysed using linear mixed effects models. ETHICS AND DISSEMINATION: Ethics approval was obtained from the University of South Australia Human Research Ethics Committee no. 204 327. Results will be disseminated through peer-reviewed journals and national and international presentations. TRIAL REGISTRATION NUMBER: NCT05267522.

Effects of weight loss through dietary intervention on pain characteristics, functional mobility, and inflammation in adults with elevated adiposity.

Background: The relationship between adiposity and pain is complex. Excess weight increases the risk for chronic musculoskeletal pain (CMP), driven by increased biomechanical load and low-grade systemic inflammation. Pain limits physical function, impacting energy balance contributing to weight gain. The primary aims of this study were to profile pain characteristics in participants with overweight or obesity and determine if weight loss through dietary-induced energy restriction, and presence of CMP, or magnitude of weight loss, was associated with changes in adiposity, pain, functional mobility, and inflammation. Methods: This was a secondary analysis of data from adults (25-65 years) with overweight or obesity (BMI 27.5-34.9 kg/m2) enrolled in a 3-month, 30% energy-restricted dietary intervention to induce weight loss (January 2019-March 2021). Anthropometric measures (weight, waist circumference and fat mass), pain prevalence, pain severity (McGill Pain Questionnaire, MPQ), pain intensity (Visual Analog Scale, VAS), functional mobility (timed up and go, TUG) and inflammation (high sensitivity C-Reactive Protein, hsCRP) were assessed at baseline and 3-months. Results: One hundred and ten participants completed the intervention and had weight and pain assessed at both baseline and 3-months. Participants lost 7.0 ± 0.3 kg, representing 7.9% ± 3.7% of body mass. At 3-months, functional mobility improved (TUG -0.2 ± 0.1 s, 95% CI -0.3, -0.1), but there was no change in hsCRP. Compared to baseline, fewer participants reported CMP at 3-months (n = 56, 51% to n = 27, 25%, p < 0.001) and presence of multisite pain decreased from 22.7% to 10.9% (p < 0.001). Improvements in anthropometric measures and functional mobility did not differ between those presenting with or without CMP at baseline. Improvements in pain were not related to the magnitude of weight loss. Conclusion: Weight loss was effective in reducing pain prevalence and improving functional mobility, emphasizing the importance of considering weight-loss as a key component of pain management. Clinical trial registration: identifier, ACTRN12618001861246.

Almonds vs. carbohydrate snacks in an energy-restricted diet: Weight and cardiometabolic outcomes from a randomized trial.

OBJECTIVE: This study evaluated weight and cardiometabolic outcomes after a 3-month energy-restricted diet (-30%) containing almonds (almond-enriched diet [AED]) or containing carbohydrate-rich snacks (nut-free control diet [NFD]) (Phase 1), followed by 6 months of weight maintenance (Phase 2). METHODS: Participants (25-65 years old) with overweight or obesity (BMI 27.5-34.9 kg/m2 ) were randomly allocated to AED (n = 68) or NFD (n = 72). RESULTS: Both groups lost weight during Phase 1 (p < 0.001) (mean [SE], -7.0 [0.5] kg AED vs. -7.0 [0.5] kg NFD, p = 0.858) and Phase 2 (p = 0.009) (-1.1 [0.5] kg AED vs. -1.3 [0.6] NFD, p = 0.756), with improvements in percentage lean mass after Phase 2 (4.8% [0.3%], p < 0.001). Reductions occurred in fasting glucose (-0.2 [0.07] mmol/L, p = 0.003), insulin (-8.1 [4.0] pmol/L, p = 0.036), blood pressure (-4.9 [0.8] mm Hg systolic, -5.0 [0.5] mm Hg diastolic, p < 0.001), total cholesterol (-0.3 [0.1] mmol/L), low-density lipoprotein (LDL) (-0.2 [0.1] mmol/L), very low-density lipoprotein (-0.1 [0.03] mmol/L), and triglycerides (-0.3 [0.06] mmol/L) (all p < 0.001), and high-density lipoprotein increased (0.1 [0.02] mmol/L, p = 0.011) by the end of Phase 2 in both groups. There were group by time interactions for lipoprotein particle concentrations: very small triglyceride-rich (-31.0 [7.7] nmol/L AED vs. -4.8 [7.9] nmol/L NFD, p = 0.007), small LDL (-109.3 [40.5] nmol/L AED vs. -20.7 [41.6] nmol/L NFD, p = 0.017), and medium LDL (-24.4 [43.4] nmol/L AED vs. -130.5 [44.4] nmol/L NFD, p = 0.045). CONCLUSIONS: An energy-restricted AED resulted in weight loss and weight loss maintenance comparable to an energy-restricted NFD, and both diets supported cardiometabolic health. The AED resulted in greater improvements in some lipoprotein subfractions, which may enhance reductions in cardiovascular risk.

Effect of a 12-Week Almond-Enriched Diet on Biomarkers of Cognitive Performance, Mood, and Cardiometabolic Health in Older Overweight Adults.

Long term nut consumption is associated with reduced risk of coronary heart disease and better cognitive function. This study examined supplementing habitual diets with almonds or carbohydrate-rich snack foods (providing 15% energy) on biomarkers of cardiovascular and metabolic health, mood and cognitive performance. Participants (overweight/obese, 50-80 years) were randomised to an almond-enriched diet (AED) or isocaloric nut-free diet (NFD) for 12 weeks. Body weight, blood lipids, glucose, insulin, blood pressure (BP), arterial stiffness, cell adhesions molecules, C reactive protein (CRP), mood, and cognitive performance (working memory primary outcome), dietary profiles and energy intake/expenditure were measured at baseline and Week 12 in 128 participants (n = 63 AED, n = 65 NFD). Compared with NFD, AED was associated with altered macro and micronutrient profiles, but no differences in energy intake or expenditure. The AED significantly reduced triglycerides and SBP but there were no other changes in cardiometabolic biomarkers, mood, or cognitive performance. The inclusion of almonds in the diet improves aspects of cardiometabolic health without affecting cognitive performance or mood in overweight/obese adults.

Study protocol for a 9-month randomised controlled trial assessing the effects of almonds versus carbohydrate-rich snack foods on weight loss and weight maintenance.

INTRODUCTION: Epidemiological studies indicate an inverse association between nut consumption and body mass index (BMI). However, clinical trials evaluating the effects of nut consumption compared with a nut-free diet on adiposity have reported mixed findings with some studies reporting greater weight loss and others reporting no weight change. This paper describes the rationale and detailed protocol for a randomised controlled trial assessing whether the inclusion of almonds or carbohydrate-rich snacks in an otherwise nut-free energy-restricted diet will promote weight loss during 3 months of energy restriction and limit weight regain during 6 months of weight maintenance. METHODS AND ANALYSIS: One hundred and thirty-four adults aged 25-65 years with a BMI of 27.5-34.9 kg/m2 will be recruited and randomly allocated to either the almond-enriched diet (AED) (15% energy from almonds) or a nut-free control diet (NFD) (15% energy from carbohydrate-rich snack foods). Study snack foods will be provided. Weight loss will be achieved through a 30% energy restriction over 3 months, and weight maintenance will be encouraged for 6 months by increasing overall energy intake by ~120-180 kcal/day (~500-750kJ/day) as required. Food will be self-selected, based on recommendations from the study dietitian. Body composition, resting energy expenditure, total daily energy expenditure (via doubly labelled water), physical activity, appetite regulation, cardiometabolic health, gut microbiome, liver health, inflammatory factors, eating behaviours, mood and personality, functional mobility and pain, quality of life and sleep patterns will be measured throughout the 9-month trial. The effects of intervention on the outcome measures over time will be analysed using random effects mixed models, with treatment (AED or NFD) and time (baseline, 3 months and 9 months) being the between and within factors, respectively in the analysis. ETHICS AND DISSEMINATION: Ethics approval was obtained from the University of South Australia Human Research Ethics Committee (201436). Results from this trial will be disseminated through publication in peer-reviewed journals, national and international presentations. TRIAL REGISTRATION NUMBER: Australian New Zealand Clinical Trials Registry (ACTRN12618001861246).

Effects of almond, dried grape and dried cranberry consumption on endurance exercise performance, recovery and psychomotor speed: protocol of a randomised controlled trial.

BACKGROUND: Foods rich in nutrients, such as nitrate, nitrite, L-arginine and polyphenols, can promote the synthesis of nitric oxide (NO), which may induce ergogenic effects on endurance exercise performance. Thus, consuming foods rich in these components, such as almonds, dried grapes and dried cranberries (AGC), may improve athletic performance. Additionally, the antioxidant properties of these foods may reduce oxidative damage induced by intense exercise, thus improving recovery and reducing fatigue from strenuous physical training. Improvements in NO synthesis may also promote cerebral blood flow, which may improve cognitive function. METHODS AND ANALYSIS: Ninety-six trained male cyclists or triathletes will be randomised to consume ~2550 kJ of either a mixture of AGC or a comparator snack food (oat bar) for 4 weeks during an overreaching endurance training protocol comprised of a 2-week heavy training phase, followed by a 2-week taper. The primary outcome is endurance exercise performance (5 min time-trial performance) and secondary outcomes include markers of NO synthesis (plasma and urinary nitrites and nitrates), muscle damage (serum creatine kinase and lactate dehydrogenase), oxidative stress (F2-isoprostanes), endurance exercise function (exercise efficiency, submaximal oxygen consumption and substrate utilisation), markers of internal training load (subjective well-being, rating of perceived exertion, maximal rate of heart rate increase and peak heart rate) and psychomotor speed (choice reaction time). CONCLUSION: This study will evaluate whether consuming AGC improves endurance exercise performance, recovery and psychomotor speed across an endurance training programme, and evaluate the mechanisms responsible for any improvement. TRIAL REGISTRATION NUMBER: ACTRN12618000360213.

LONG R3IGF-I as a more potent alternative to insulin in serum-free culture of HEK293 cells.

LONG R3IGF-I, an analogue of insulin-like growth factor (IGF)-I, was specifically engineered for use in biopharmaceutical protein production in mammalian cells. LONG R3IGF-I is capable of supporting the growth and survival of Chinese hamster ovary cells in serum-free media at concentrations at least 200-fold lower than required for insulin. LONG R3IGF-I also acts as a more potent growth and survival factor than either insulin or native IGF-I in SF culture of human embryonic kidney (HEK293) cells. To investigate the basis of the enhanced potency of LONG R3IGF-I we have examined the mechanism of action of these mitogens in HEK293 cells. All mitogens tested were found to activate the Type I IGF receptor (IGF-IR) and insulin receptor (IR) in a dose-responsive manner. However, the level of activation of both receptors after stimulation with LONG R3IGF-I, at lower concentrations, was greater than with either insulin or IGF-I. The greater potency of LONG R3IGF-I in activating the IR, despite having a low affinity for IRs, suggests the presence of heterotetrameric IGF-IR/IR dimers. Interestingly, the decrease in IGF-IR activation at higher concentrations of LONG R3IGF-I suggests that the dose-response curve may be bell-shaped.