Background The effect of single food‐based dietary interventions on the American Heart Association's cardiovascular health score, Life's Essential 8 (LE8), is unclear. The aim of this study was to examine the effect of daily avocado intake for 26 weeks on LE8 in adults with abdominal obesity. Methods and Results An ancillary analysis including participants (n=969; aged 51±14 years) from the HAT (Habitual Diet and Avocado Trial), a randomized controlled trial, was conducted. The Avocado‐Supplemented Diet Group was provided with 1 avocado per day, and the Habitual Diet Group was instructed to maintain their usual diet. LE8 component scores (diet, physical activity, nicotine exposure, sleep health, body mass index, blood lipids, blood glucose, and blood pressure) were calculated using a modified American Heart Association algorithm. The LE8 score was calculated as the unweighted average of each component (range, 0–100 points). Between‐group differences in the 26‐week change in LE8 were assessed using general linear models. No significant between‐group difference in the 26‐week change in the LE8 score from baseline was observed (0.79 points [95% CI, −0.41 to 2.00]). However, avocado intake increased the LE8 component scores for diet (3.53 points [95% CI, 1.38–5.68]), sleep health (3.20 points [95% CI, 0.38–6.02]), and blood lipids (3.46 points [95% CI, 1.03–5.90]) compared with the Habitual Diet Group. Conclusions Intake of 1 avocado per day for 26 weeks did not significantly affect the total cardiovascular health score in US adults with abdominal obesity. However, diet quality, sleep health, and blood lipids improved with daily avocado intake. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT03528031.

Objectives: This study aimed to investigate short-term and long-term impact of avocado consumption without caloric restriction on the gut microbiota of free-living adults with abdominal obesity. Methods: The Habitual Diet and Avocado Trial (HAT) was a 26-week, multi-center, randomized, controlled trial involving 1008 individuals with abdominal obesity. Participants were randomly assigned to the Avocado Supplemented Diet Group (AVO), receiving one avocado per day, or the Habitual Diet group (HAB), maintaining their usual dietary habits. Fecal samples were collected at baseline, week 4 and week 26 from a subset of participants recruited at a University of California Los Angeles site (n = 230). Fecal microbiota was assessed with shotgun metagenomics sequencing. Alpha diversity was assessed using the Chao1 and Shannon indices; beta diversity was assessed using Bray-Curtis dissimilarity with significance determined by repeated measures permutational multivariat analysis of variance. Potential association of intervention at week 4 and 26 with alpha diversity, species and metabolic pathways was examined using linear mixed effect models. Results: Compared to the HAB group, the AVO group had higher alpha diversity by 4 weeks, which persisted through the 26-week study period. Exploratory analysis based on healthy eating index-2015 (HEI-2015) indicated that participants with a low HEI score at baseline (<= 52.7), had an increase in alpha diversity in the AVO group vs. HAB group. The AVO group had a significant change in beta diversity at week 26 compared to the HAB group. At the species level, the AVO group had significantly increased Faecalibacterium prausnitzii and Bacterium AF16_15 at week 26 compared to the HAB group. Functional analysis showed no significant difference in metabolic pathways between the HAB and AVO groups. Conclusions: Our findings document a potentially favorable effect of avocados on gut microbiota diversity. The prebiotic potential of avocados is more pronounced in individuals with a low diet quality score. This trial is registered at clinicaltrials.gov as NCT03528031 (https://clinicaltrials.gov/study/NCT03528031).

Objectives: The aim of this study was to examine the adherence, changes in weight, and, waist circumference associated with the daily consumption of a culturally preferred food, namely an avocado, among Hispanic/Latina females in the Habitual Diet and Avocado Trial (HAT). Methods: HAT was a multisite, randomized controlled trial conducted between 2018 and 2020. Participants in the Avocado-Supplemented Diet Group were provided with and instructed to consume one avocado/day (~2.2 servings) for 6 months; participants in the Habitual Diet Group were instructed to follow their usual diet and limit intake to ≤2 avocados/month. Avocado consumption was assessed using three random 24 h dietary recalls administered by dietitians. This analysis focused on women who self-identified as Hispanic/Latina. Results: Within HAT, 158 females self-identified as Hispanic/Latina (median age: 42 years, IQR: 36–54). Across the dietary recalls, the Avocado-Supplemented Group (n = 80) consumed 1.9–2.1 avocado servings/day; the Habitual Diet Group (n = 78) consumed 0.04–0.09 servings/day (p < 0.001). The weight and waist circumference measurements were similar between groups. Hispanic/Latina females remained adherent to daily avocado consumption for the 6-month study period, without a significant change in their body weight or waist circumference measurements. Conclusions: Integrating a culturally preferred food into a dietary intervention enhanced adherence amongst Latina adults, with no impact significant impact on body composition.

Context Total fasting non-esterified fatty acid (NEFA) levels have been associated with mortality. The corresponding associations with NEFA levels following an oral glucose tolerance test (OGTT) and with individual fasting NEFA species are unclear.Objective We evaluated the associations of post-load NEFA, fasting subclasses and individual NEFA with mortality.Methods The Cardiovascular Health Study is a population-based cohort study of community-dwelling adults over 64 years of age from 4 US communities that began in 1989-1990. Participants had total NEFA measured enzymatically before and 2 hours after an OGTT from archived serum samples collected in 1996-1997. Fasting individual NEFA were also measured using gas chromatography. Cox proportional hazard models were used to evaluate adjusted hazard ratios (aHR) for mortality associated with fasting and post-load total NEFA, and fasting individual and fatty acid subclasses (saturated, monounsaturated, n-3 and n-6 polyunsaturated, and trans).Results The final population included 1996 participants (mean age 78 years; 60.5% female). Over a median 11-year follow-up period, 1678 participants died. Total fasting NEFA was associated with higher risk of all-cause mortality (aHR per SD: 1.17, 95% CI [1.10-1.23]). Total post-load NEFA was not associated with mortality. Among subclasses, only monounsaturated fatty acid (MUFA) was associated with total mortality (aHR 1.24, 95% CI [1.09-1.41]). For individual NEFAs, nervonic acid (aHR 1.06, 95% CI [1.01-1.12]), petroselaidic acid (aHR 1.21, 95% CI [1.03-1.42]), and eicosapentaenoic acid (aHR 0.90, 95% CI [0.82-0.99]) were associated with all-cause mortality.Conclusion Individual fasting NEFAs represent attractive candidates for medical and public health interventions aimed at improving survivorship in older adults and should be investigated further.

Diet is a potentially modifiable neurodegenerative disease risk factor. We studied the effects of a typical Western diet (WD; high in refined carbohydrates, cholesterol and saturated fat), relative to a heart-healthy diet (HHD; high in unrefined carbohydrates, polyunsaturated fat and fiber, and low in cholesterol) on brain microvessel transcriptomics and brain metabolomics of the temporal region in Ossabaw minipigs. Thirty-two pigs (16 male and 16 females) were fed a WD or HHD starting at the age of 4 months for a period of 6 months. The WD and HHD were isocaloric and had a similar macronutrient content but differed in macronutrient quality. Within each dietary group, half of the pigs also received atorvastatin. Relative to HHD-fed pigs, WD-fed pigs had 175 genes differentially expressed (fold change > 1.3, FDR < 0.05) by diet, 46 upregulated and 129 downregulated. Gene Set Enrichment Analysis identified 22 gene sets enriched in WD-fed pigs, comprising pathways related to inflammation, angiogenesis, and apoptosis, and 53 gene sets enriched in the HHD-fed pigs, including cell energetics, neurotransmission, and inflammation resolution pathways. Metabolite analysis showed enrichment in arginine, tyrosine, and lysine in WD-fed pigs, and ergothioneine and S-adenosyl methionine in HHD-fed pigs. Atorvastatin treatment did not affect gene expression. These results suggest a likely contribution of diet to brain pathologies characterized by neuroinflammation and neurodegeneration.

BACKGROUND:Avocado intake improves dietary fat quality, but the subsequent impact on red blood cell (RBC) saturated (SFA), monounsaturated (MUFA), polyunsaturated (PUFA), and trans-fatty acid (TFA) composition and association with cardiometabolic health, has not been elucidated. OBJECTIVES:To compare the effect of consuming 1 avocado/d relative to habitual diet (HAB) on RBC-FA profiles, and their association with visceral adiposity and cardiometabolic risk factors (CMRFs) in individuals with abdominal obesity. METHODS:RBC-FA profiling at baseline, 3- and 6 mo was conducted in participants (n = 994) from the Habitual Diet and Avocado Trial (HAT). HAT was a multisite, free-living, parallel-arm intervention study in which participants were randomly assigned to either the avocado-supplemented group (AVO, usual diet with 1 avocado/d) or the HAB group (usual diet with limited avocado intake) for 6 mo. Changes in RBC-FA profiles, a secondary outcome measure, were determined within and between groups using linear regression and mixed effect models, adjusting for age, sex, BMI, clinical site, smoking status, and percentage of energy intake from fat at baseline. The association between changes in RBC-FAs with visceral adiposity measures and CMRFs was assessed after covariate and False Discovery Rate (FDR <0.05) adjustment. RESULTS:No major differences in RBC-FA profiles were observed between groups, with the exception of MUFA cis-vaccenic [18:1n-7c], which was significantly higher in AVO (β: 0.11 [0.05, 0.17]) compared with the HAB (β: 0.03 [-0.03, 0.08]) participants. In the HAB but not AVO group, increases in MUFA cis (18:1n-7c, oleic [18;1n-9c], erucic [22:1n-9c]) and MUFA trans (palmitelaidic [16:1n-7t], vaccenic [18:1n-7t], elaidic [18:1n-9t], and petroselaidic [18;1n-10-12t), as well as PUFA γ-linolenic [18:3n-6], dihomo-γ-linolenic [20:3n-6], arachidonic [20:4n-6], and α-linolenic [18:3n-3] were associated with unfavorable changes in visceral adiposity measures, lipid profiles, glucose, insulin and high sensitivity C-reactive protein concentrations. CONCLUSIONS:Daily avocado intake over 6-mo modified RBC-MUFA composition, notably 18:1n-7c, and potentially mitigated some of the unfavorable individual RBC-FA-CMRF associations observed over time in the HAB group. This trial was registered at https://clinicaltrials.gov/study as NCT03528031.

Objective Inflammation worsens joint destruction in osteoarthritis (OA) and aggravates pain. Although n‐3 fatty acids reduce inflammation, different n‐3 fatty acids have different effects on inflammation and clinical outcomes, with eicosapentaenoic acid (EPA) having the strongest effect. We examined whether specific essential fatty acid levels affected the development of OA. Methods We studied participants from the Multicenter Osteoarthritis Study (MOST) at risk of developing knee OA. As part of MOST, participants were asked repeatedly about knee pain, and knee radiographs and magnetic resonance images (MRIs) were obtained. Using baseline fasting samples, we analyzed serum fatty acids with standard assays. After excluding participants with baseline OA, we defined two sets of cases based on their status through 60 months’ follow‐up: those developing incident radiographic OA and those developing incident symptomatic OA (knee pain and radiographic OA). Controls did not develop these outcomes. Additionally, we examined worsening of MRI cartilage damage and synovitis and worsening knee pain and evaluated the number of hand joints affected by nodules. In regression models, we tested the association of each OA outcome with levels of specific n‐3 and n‐6 fatty acids, adjusting for age, sex, body mass index, education, physical activity, race, baseline pain, smoking, statin use, and depressive symptoms. Results We studied 363 cases with incident symptomatic knee OA and 295 with incident radiographic knee OA. The mean age was 62 years (59% women). We found no associations of specific n‐3 fatty acid levels, including EPA, or of n‐6 fatty acid levels with incident OA (eg, for incident symptomatic knee OA, the odds ratio per SD increase in EPA was 1.0 [95% confidence interval 0.87–1.17]). Results for other OA outcomes also failed to suggest a protective effect of specific n‐3 fatty acids with OA outcomes. Conclusion We found no association of serum levels of EPA or of other specific n‐3 fatty acids or n‐6 fatty acids with risk of incident knee OA or other OA outcomes.

The rate of cardiovascular disease (CVD) death is higher in men than women before age 50 y, but the gap between sexes significantly narrows after menopause. Lipid mediators derived from EPA, DHA and AA play a role in inflammation and CVD. The aim of our study was to assess whether plasma concentrations of these lipid mediators differ between postmenopausal women and men. Twelve postmenopausal women and 9 men with low-grade chronic inflammation completed a randomized, double-blind, crossover study consisting of a 4-week lead-in placebo phase (3 g/d high-oleic acid sunflower oil) followed by randomization to either 3 g/d DHA or 3 g/d EPA for 10 weeks and crossover for additional 10 weeks, separated by a washout phase. Plasma phospholipid content of EPA, DHA and AA and plasma concentrations of their derived lipid mediators were measured at the end of the placebo lead-in phase (baseline) and the DHA and EPA supplementation phases. There were no sex differences in plasma phospholipid EPA, DHA and AA at baseline and after DHA and EPA supplementation. However, plasma concentrations of lipid mediators derived from EPA, DHA and AA via 15-lipoxygenase were lower in postmenopausal women than men, especially after supplementation. Sex differences in EPA- and DHA-derived lipid mediators with anti-inflammatory and pro-resolving actions may partly explain the faster rise in CVD in postmenopausal women than age-matched men.

Background: Dietary stearic acid (18:0), a saturated fatty acid (SFA) commonly present in Western diets, has an LDL-C lowering effect compared to shorter chain SFAs such as palmitic acid (16:0), and a similar effect compared to oleic acid (18:1). However, the underlying mechanisms remain unclear. Hypothesis: We tested the hypothesis that the hypocholesterolemic effect of dietary 18:0 and 18:1 relative to 16:0 is modulated by alterations in cholesterol and bile acid (BA) metabolism. Methods: This secondary analysis used archived plasma and fecal samples from a randomized crossover feeding study (N=20 mildly hypercholesteremic postmenopausal women, 64±7 years, BMI 26.4±3.4kg/m 2 ). Participants consumed each of 3 isocaloric diets enriched in either 18:0, 16:0 or 18:1 for five weeks with a 2-week washout. Primary (P) and secondary (S) BAs, and their conjugates were measured in fecal, fasting and non-fasting (NF) plasma samples using the Biocrates MxP Quant 500 kit and Quadrupole Time-of-Flight mass spectrometry. Fasting and NF plasma cholesterol synthesis (lathosterol) and absorption (-sitosterol) markerswere quantified using gas chromatography. Mixed-effect and generalized linear mixed models were used to test the difference in outcome measures among diets, with Tukey-Kramer post hoc comparison. Spearman correlation coefficients with FDR adjustment was calculated between BA, cholesterol synthesis/absorption markers, and CVD risk factors. Results: Compared to the 16:0 diet, consumption of the 18:0 diet resulted in significantly lower fasting and NF plasma lathosterol (-22%); higher -sitosterol (19%); higher fecal PBAs (31%) and lower fecal SBAs (-17%) concentrations. Plasma PBAs were significantly lower in the fasted state (-34%), but higher in the NF state (21%; 18:0 vs. 16:0). Interestingly, conjugated PBA and SBA concentrations in the NF state were significantly higher after participants consumed the 18:0 compared to the 18:1 diet (all p < 0.05). Plasma NF PBAs were positively associated with -sitosterol (r=0.56, p < 0.05), while plasma fasting PBAs were negatively associated with lathosterol (r=-0.58, p < 0.05). Conjugated PBA and SBA concentrations were negatively associated with LDL-C, hsCRP, E-selectin and insulin concentrations in the fasted but not NF state. Conclusion: The favorable effects of 18:0 on CVD risk factors may be modulated, in part, by changes in BA and cholesterol metabolism, with distinct effects in the fasted and NF states.

Background: Total fasting non-esterified fatty acid (NEFA) levels have been associated with total and cause-specific mortality in older adults, but the corresponding associations with NEFA levels following the metabolic challenge of an oral glucose tolerance test (OGTT) and with individual fasting NEFA species have not been quantified Methods: We studied 1996 participants from the Cardiovascular Health Study with a mean age of 78 years who had total serum NEFA measured enzymatically before and two hours after an OGTT and fasting individual NEFA measured using gas chromatography in samples collected in 1996-1997. 1678 participants died over a median follow-up of 11 years. Multivariable Cox proportional hazard models were used to evaluate adjusted hazard ratios (aHR) for mortality associated with standard deviation increments of fasting and post-load total NEFA and fasting individual and subclasses (saturated, monounsaturated, n-3 and n-6 polyunsaturated, and trans) Results: Fasting NEFA were associated with an increased risk of all-cause mortality (aHR 1.17, 95% CI 1.10 - 1.23). In contrast, levels of post-load NEFA were not associated with mortality (aHR 0.98, 95% CI 0.93 - 1.03). Among NEFA subclasses, only monounsaturated fatty acid (MUFA) were associated with total mortality (aHR 1.24, 95% CI 1.09 - 1.41). In cause-specific analyses, MUFA were associated with higher risk of death from dementia, cardiovascular, respiratory, infectious and kidney disease. For individual NEFAs, nervonic acid (aHR 1.06, 95% CI 1.01 - 1.12) and petroselaidic acid (aHR 1.21, 95% CI 1.03 - 1.42) were associated with higher risk of all-cause mortality whereas eicosapentaenoic acid (aHR 0.90, 95% CI 0.82 - 0.99) was associated with lower risk Conclusion: Among community-dwelling older adults, total fasting but not post-load NEFA were associated with higher risk of death. Among subclasses and individual species, MUFA, nervonic, and petroselaidic acids were associated with higher risk, and eicosapentanoic acid with lower risk, of mortality.

Background Abdominal obesity is associated with endothelial dysfunction and poorer vascular health. Avocado consumption improves postprandial endothelial function; however, the longer‐term effects remain unclear. It was hypothesized that the daily addition of 1 avocado to a habitual diet for 6 months would improve flow‐mediated dilation (FMD) and carotid–femoral pulse wave velocity in individuals with abdominal obesity (waist circumference ≥35 in for women, ≥40 in for men), compared with a habitual diet low in avocados. Methods and Results HAT (Habitual Diet and Avocado Trial) was a multicenter, randomized, controlled, parallel‐arm study that investigated the health effects of adding 1 avocado per day to a habitual diet in individuals with abdominal obesity. At the Pennsylvania State University, University Park study center (n=134; age, 50 ± 13 years; women, 78%; body mass index, 32.6 ± 4.8 kg/m2), markers of vascular function were measured, including endothelial function, assessed via brachial artery flow‐mediated dilation, and arterial stiffness, assessed via carotid–femoral pulse wave velocity. Between‐group differences in 6‐month change in flow‐mediated dilation and carotid–femoral pulse wave velocity were assessed using independent t tests. Prespecified subgroup analyses were conducted using linear regression. No significant between‐group differences in flow‐mediated dilation (mean difference=−0.62% [95% CI, −1.70 to 0.46]) or carotid–femoral pulse wave velocity (0.25 m/s [95% CI, −0.13 to 0.63]) were observed. Results of the subgroup analyses were consistent with the primary analyses. Conclusions Longer‐term consumption of 1 avocado per day as part of a habitual diet did not improve measures of vascular function compared with a habitual diet low in avocados in individuals with abdominal obesity. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT03528031.

When subject to damage or stress, cells develop responses in order to maintain tissue homeostasis. Two such decisions are ferroptosis and cellular senescence, but how cells decide between these outcomes remains unclear. Here we show that senescent cells increase levels of multiple membrane bound polyunsaturated fatty acids (PUFAs), but a specific PUFA, dihomo−gamma−linolenic acid (DGLA, 20:3−ω3) is reduced. Exogenous repletion of DGLA or inhibition of the enzyme that metabolizes DGLA, delta−5−desaturase, instead results in cell death by ferroptosis. Senescent cells also show elevated levels of other ferroptosis sensitizers, including labile iron and expression of lipoxygenases − but also increase Gpx4 levels. Oral administration of exogenous DGLA lowers senescent cell burden in aged mice and improves age−related functional outcomes. Finally, obese humans with lowered DGLA desaturation rates showed lower markers of adipose tissue senescence. Together, our data implicate DGLA and its desaturation as a major driver of decisions between senescence and ferroptosis. ### Competing Interest Statement CW is an inventor on patents related to the elimination of senescent cells using DGLA, D5D inhibitors, and other ferroptosis inducers. The content is the sole responsibility of the authors and does not necessarily represent the official views of the USDA.

Background: Few clinical trials have evaluated diet quality change as a predictor of intervention effectiveness. Objectives: The aim of this study was to examine changes in the Healthy Eating Index (HEI)-2015 after a food -based intervention, and assess the associations between HEI-2015 change and intervention effects on cardiometabolic risk-related outcomes. Methods: The Habitual Diet and Avocado Trial was a 26 -wk, multicenter, randomized, controlled parallel -arm study. Participants were 1008 individuals aged >= 25 y with abdominal obesity (females >= 35 inches; males >= 40 inches). The avocado -supplemented diet group was provided 1 avocado per day, and the habitual diet group maintained their usual diet. Change in diet quality was assessed using the HEI-2015 from a single 24-h recall conducted at 4 time points. Mixed models were used for analysis. Results: The avocado -supplemented diet group had a greater increase in the HEI-2015 (4.74 points; 95% CI: 2.93, 6.55) at 26 wk than the habitual diet group. Compared with the habitual diet group, the avocado -supplemented diet group had greater increases in the following HEI-2015 components from baseline: total vegetables (0.99 points; 95% CI: 0.77, 1.21), fatty acid ratio (2.25 points; 95% CI: 1.74, 2.77), sodium (1.03 points; 95% CI: 0.52, 1.55), refined grains (0.82 points; 95% CI: 0.32, 1.31), and added sugars (0.84 points; 95% CI: 0.49, 1.19). No differences in HEI-2015 improvements were observed by race, ethnicity, study site, body mass index, or age category. In the avocado -supplemented diet compared with the habitual diet group, the HEI-2015 increased in females (6.50 points; 95% CI: 4.39, 8.62) but not in males (0.02 points; 95% CI: -3.44, 3.48). Median HEI-2015 change was not associated with intervention -related changes in cardiometabolic disease risk factors. Conclusions: Intake of 1 avocado per day for 26 wk in adults with abdominal obesity increased adherence to the Dietary Guidelines for Americans. Changes in diet quality did not predict changes in risk factors for cardiometabolic disease. This trial was registered at clinicaltrials.gov as NCT03528031 (https://clinicaltrials.gov/study/NCT03528031).

BACKGROUND:Recent data indicate considerable variability in response to very long chain omega-3 fatty acid supplementation on cardiovascular disease risk. This inconsistency may be due to differential effects of EPA vs DHA and/or sex-specific responses. METHODS:Sixteen subjects (eight men and eight women) 50-75 y and with low-grade chronic inflammation participated in a randomized controlled crossover trial comparing 3 g/d EPA, 3 g/d DHA, and placebo (3 g/d high oleic acid sunflower oil). Blood monocytes were isolated at the end of each phase for RNA-sequencing. RESULTS:Sex dimorphism in monocyte gene expression was observed, therefore, data for men and women were analyzed separately. 1088 genes were differentially expressed in men and 997 in women (p < 0.05). In both men and women, EPA and DHA repressed genes involved in protein turnover and mitochondrial energy metabolism, relative to placebo. In men only, EPA and DHA upregulated genes related to wound healing and PPARα activation. In women only, EPA and DHA activated genes related to ER stress response. Relative to DHA, EPA resulted in lower expression of genes involved in inflammatory processes in men, and lower expression of genes involved in ER stress response in women. CONCLUSIONS:EPA and DHA supplementation elicited both similar and differential effects on monocyte transcriptome, some of which were sex specific. The observed variability in response to EPA and DHA in men and women could in part explain the conflicting results from previous cardiovascular clinical trials using omega-3 fatty acids.