Tuccinardi, D., O.M. Farr, J. Upadhyay, S.M. Oussaada, M.I. Klapa, M. Candela, S. Rampelli, S. Lehoux, I. Lázaro, A. Sala‐Vila, P. Brigidi, R.D. Cummings, C.S. Mantzoros, 2019. Mechanisms underlying the cardiometabolic protective effect of walnut consumption in obese subjects: a cross‐over, randomized, double‐blinded, controlled inpatient physiology study. Diabetes Obes Metab. 21(9):2086-2095.
Aims: To assess the effects of walnuts on cardiometabolic outcomes in obese subjects and to explore underlying mechanisms using novel methods including metabolomic, lipidomic, glycomic, and microbiome analysis integrated with lipid particle fractionation, appetite-regulating hormones and hemodynamic measurements. Materials and Methods: 10 obese subjects were enrolled in this cross-over, randomized, double-blind, placebo-controlled clinical trial. Patients participated in two 5-day inpatient stays during which they consumed a smoothie containing 48g walnuts or a macronutrient-matched placebo smoothie without nuts, with a one-month washout period between the two visits. Results: Walnut consumption improved aspects of the lipid profile, i.e. reduced fasting small and dense LDL particles (p<.02) and increased postprandial large HDL particles (p<.01). Lipoprotein Insulin Resistance Score, glucose and insulin AUC decreased significantly after walnut consumption (p<.01, p<.02, p<.04, respectively). Consuming walnuts significantly increased 10 N-glycans, with 8 of them carrying a fucose core. Lipidomic analysis showed a robust reduction in harmful ceramides, hexosylceramides and sphingomyelins, which have been shown to mediate effects on cardiometabolic risk. Peptide YY AUC significantly increased after walnut consumption (p<.03). No major significant changes in hemodynamic, metabolomic analysis or in host health-promoting bacteria such as Faecalibacterium were found. Conclusions: These data provide a more comprehensive mechanistic perspective of the effect of dietary walnut consumption on cardiometabolic parameters. Lipidomic and lipid nuclear magnetic resonance spectroscopy analysis showed an early but significant reduction in ceramides and other atherogenic lipids with walnut consumption that may explain the longer-term benefits of walnuts on insulin resistance, cardiovascular risk and mortality.
Tindall, A.M., K.S. Petersen, A.C. Skulas-Ray, C.K. Richter, D.N. Proctor, P.M. Kris-Etherton, 2019. Replacing saturated fat with walnuts or vegetable oils improves central blood pressure and serum lipids in adults at risk for cardiovascular disease: a randomized controlled-feeding trial. J Am Heart Assoc. 8(9):e011512. doi: 10.1161/JAHA.118.011512
Background: Walnuts have beneficial effects on cardiovascular risk factors, but it is unclear whether these effects are attributable to the fatty acid ( FA ) content, including α-linolenic acid ( ALA ), and/or bioactives. Methods and Results: A randomized, controlled, 3-period, crossover, feeding trial was conducted in individuals at risk for cardiovascular disease (n=45). Following a 2-week standard Western diet run-in (12% saturated FAs [ SFA ], 7% polyunsaturated FAs, 12% monounsaturated FAs), participants consumed 3 isocaloric weight-maintenance diets for 6 weeks each: a walnut diet ( WD ; 7% SFA , 16% polyunsaturated FAs, 3% ALA , 9% monounsaturated FAs); a walnut FA -matched diet; and an oleic acid-replaced- ALA diet (7% SFA , 14% polyunsaturated FAs, 0.5% ALA , 12% monounsaturated FAs), which substituted the amount of ALA from walnuts in the WD with oleic acid. This design enabled evaluation of the effects of whole walnuts versus constituent components. The primary end point, central systolic blood pressure, was unchanged, and there were no significant changes in arterial stiffness. There was a treatment effect ( P=0.04) for central diastolic blood pressure; there was a greater change following the WD versus the oleic acid-replaced-ALA diet (-1.78±1.0 versus 0.15±0.7 mm Hg, P=0.04). There were no differences between the WD and the walnut fatty acid-matched diet (-0.22±0.8 mm Hg, P=0.20) or the walnut FA-matched and oleic acid-replaced-ALA diets ( P=0.74). The WD significantly lowered brachial and central mean arterial pressure. All diets lowered total cholesterol, LDL (low-density lipoprotein) cholesterol, HDL (high-density lipoprotein) cholesterol, and non- HDL cholesterol. Conclusions: Cardiovascular benefits occurred with all moderate-fat, high-unsaturated-fat diets. As part of a low- SFA diet, the greater improvement in central diastolic blood pressure following the WD versus the oleic acid-replaced-ALA diet indicates benefits of walnuts as a whole-food replacement for SFA.
Tindall, A.M., C.J. McLimans, K.S. Petersen, P.M. Kris-Etherton, R. Lamendella, 2019. Walnuts and vegetable oils containing oleic acid differentially affect the gut microbiota and associations with cardiovascular risk factors: Follow-up of a randomized, controlled, feeding trial in adults at risk for cardiovascular disease. J Nutr. 150(4):806-817.
Background: It is unclear whether the favorable effects of walnuts on the gut microbiota are attributable to the fatty acids, including α-linolenic acid (ALA), and/or the bioactive compounds and fiber. Objective: This study examined between-diet gut bacterial differences in individuals at increased cardiovascular risk following diets that replace SFAs with walnuts or vegetable oils.Methods: Forty-two adults at cardiovascular risk were included in a randomized, crossover, controlled-feeding trial that provided a 2-wk standard Western diet (SWD) run-in and three 6-wk isocaloric study diets: a diet containing whole walnuts (WD; 57-99 g/d walnuts; 2.7% ALA), a fatty acid-matched diet devoid of walnuts (walnut fatty acid-matched diet; WFMD; 2.6% ALA), and a diet replacing ALA with oleic acid without walnuts (oleic acid replaces ALA diet; ORAD; 0.4% ALA). Fecal samples were collected following the run-in and study diets to assess gut microbiota with 16S rRNA sequencing and Qiime2 for amplicon sequence variant picking. Results: Subjects had elevated BMI (30 ± 1 kg/m2), blood pressure (121 ± 2/77 ± 1 mmHg), and LDL cholesterol (120 ± 5 mg/dL). Following the WD, Roseburia [relative abundance (RA) = 4.2%, linear discriminant analysis (LDA) = 4], Eubacterium eligensgroup (RA = 1.4%, LDA = 4), LachnospiraceaeUCG001 (RA = 1.2%, LDA = 3.2), Lachnospiraceae UCG004 (RA = 1.0%, LDA = 3), and Leuconostocaceae (RA = 0.03%, LDA = 2.8) were most abundant relative to taxa in the SWD (P ≤ 0.05 for all). The WD was also enriched in Gordonibacter relative to the WFMD. Roseburia (3.6%, LDA = 4) and Eubacterium eligensgroup (RA = 1.5%, LDA = 3.4) were abundant following the WFMD, and Clostridialesvadin BB60group (RA = 0.3%, LDA = 2) and gutmetagenome (RA = 0.2%, LDA = 2) were most abundant following the ORAD relative to the SWD (P ≤ 0.05 for all). Lachnospiraceae were inversely correlated with blood pressure and lipid/lipoprotein measurements following the WD. Conclusions: The results indicate similar enrichment of Roseburia following the WD and WFMD, which could be explained by the fatty acid composition. Gordonibacter enrichment and the inverse association between Lachnospiraceae and cardiovascular risk factors following the WD suggest that the gut microbiota may contribute to the health benefits of walnut consumption in adults at cardiovascular risk. This trial was registered at clinicaltrials.gov as NCT02210767.
Shin, P.K., Y. Zoh, J. Choi, M.S. Kim,Y. Kim, S.W. Choi, 2019. Walnut phenolic extracts reduce telomere length and telomerase activity in a colon cancer stem cell model. Nutr Res Pract. 13(1):58-63.
Background/Objectives: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. Materials/Methods: CD133+CD44+ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and 40 µg/mL for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA (ddCt). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA (dCt). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. Results: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner (5.16 ± 0.13 at 0 µg/mL, 4.79 ± 0.12 at 10 µg/mL 3.24 ± 0.08 at 20 µg/mL and 3.99 ± 0.09 at 40 µg/mL; P = 0.0276). Telomerase activities concurrently decreased with telomere length (1.47 ± 0.04, 1.09 ± 0.01, 0.76 ± 0.08, and 0.88 ± 0.06; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. Conclusions: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.
Senevirathne, A., E. Neale, G. Peoples, L. Tapsell, 2019. Relationship between long‐chain omega‐3 polyunsaturated fatty acid intake and ankle brachial index, pulse wave velocity and resting heart rate in a sample of overweight adults: A secondary analysis of baseline data in the HealthTrack study. Nutr Diet. 76(1):95-103.
Aim: The present study aimed to explore the association between dietary long-chain omega-3 polyunsaturated fatty acid (LCn3PUFA) intake and cardiovascular risk indicators (ankle brachial index, resting heart rate and brachial-ankle pulse wave velocity) in a clinical sample of overweight and obese participants volunteering for a weight loss trial. Methods: This was a secondary analysis of baseline data from the HealthTrack study (n = 351). LCn3PUFA intake was calculated via a diet history and the association with ankle brachial index, resting heart rate and brachio-ankle pulse wave velocity was explored using linear regression after controlling for covariates. Results: LCn3PUFA intake was inversely associated with ankle brachial index (R(2) change = 0.021, F change (1, 339) = 8.864, P < 0.05) and resting heart rate (R(2) change = 0.014, F change (1, 342) = 5.337, P < 0.05) but not with brachio-ankle pulse wave velocity (R(2) change = 0.001, F change (1, 339) = 0.725, P > 0.05). Conclusions: In this clinical sample of overweight adults, LCn3PUFA consumption was significantly associated with a lower resting heart rate, adding to the current evidence on the potential benefits of LCn3PUFA consumption. It also supports the value of targeting a diet rich in this nutrient when planning future dietetic approaches. Relationships with ankle brachial index and pulse wave velocity require further investigation. Future research should assess the effect of changes in dietary LCn3PUFA intake on novel cardiovascular risk indicators.
Njike, V.Y., V.C. Costales, P. Petraro, A. Annam, N. Yarandi, D.L. Katz, 2019. The resulting variation in nutrient intake with the inclusion of walnuts in the diets of adults at risk for type 2 diabetes: A randomized, controlled, crossover trial. Am J Health Promot. 33(3):430-438.
Purpose: We previously demonstrated that including walnuts in the diets of adults at risk for type 2 diabetes mellitus (T2DM) led to improved overall diet quality. This report examines the specific changes in their nutrient intake. Design: This was a randomized, controlled, modified Latin square parallel design trial with 2 treatment arms. Participants were randomized to walnut intake with, or without, dietary advice to regulate caloric intake. Within each treatment arm, they were further randomized to one of 2 sequence permutations (walnut-included/walnut-excluded or walnut-excluded/walnut-included diet), with a 3-month washout between treatment phases. Setting: Community hospital in Lower Naugatuck Valley in Connecticut. Participants: Cohort of 112 participants (31 men and 81 women) at risk for T2DM. Intervention: Participants included 56 g (366 kcal) of walnuts in their daily diets for 6 months. Measures: Nutrient intake was assessed using web-based Automated Self-Administered 24-Hour Dietary Assessment. Analysis: Data were analyzed using generalized linear models. Results: Walnut inclusion led to increased intake of total fat, calcium, magnesium, thiamin, total saturated fatty acids, and monounsaturated and polyunsaturated fatty acids (379.0 ± 90.3 g vs -136.5 ± 92.7 g, P < .01; 230.7 ± 114.2 mg vs -95.2 ± 117.4 mg, P = .05; 111.0 ± 33.9 mg vs -32.3 ± 34.9 mg, P < .01; 0.28 ± 0.2 mg vs -0.47 ± 0.2 mg, P = .02; 8.6 ± 3.4 g vs -1.1 ± 3.5 g, P =.05; 6.3 ± 3.9 g vs -6.3 ± 4.0 g, P = .03; and 25.4 ± 4.0 vs -6.6 ± 4.2 g, P < .01, respectively). Vitamin C intake decreased (-65.3 ± 55.3 mg vs 98.9 ± 56.8 mg, P = .04). Protein intake increased from baseline with the inclusion of walnuts (20.0 ± 8.8 g, P < .05). Walnut inclusion led to an increase in total calories consumed when caloric intake is not regulated. Conclusion: Including walnuts in the diets of these adults led to increased dietary intake of some nutrients associated with lower risk of developing T2DM and other cardiometabolic risk factors.
Nakanishi, M., A. Matz, C. Klemashevich, D.W. Rosenberg, 2019. Dietary walnut supplementation alters mucosal metabolite profiles during DSS-induced colonic ulceration. Nutrients. 11(5). pii: 1118. doi: 10.3390/nu11051118
Walnuts contain a complex array of natural compounds and phytochemicals that exhibit a wide range of health benefits, including protection against inflammation and colon cancer. In this study, we assess the effects of dietary supplementation with walnuts on colonic mucosal injury induced in mice by the ulcerogenic agent, dextran sodium sulfate (DSS). C57Bl/6J mice were started on the Total Western Diet supplemented with freshly-ground whole walnuts (0, 3.5, 7 and 14% g/kg) 2 weeks prior to a 5-day DSS treatment and walnut diets were continued throughout the entire experimental period. Mice were examined at 2 days or 10 days after withdrawal of DSS. In a separate study, a discovery-based metabolite profiling analysis using liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed on fecal samples and colonic mucosa following two weeks of walnut supplementation. Dietary walnut supplementation showed significant effects in the 10-day post-DSS recovery-phase study, in which the extent of ulceration was significantly reduced (7.5% vs. 0.3%, p < 0.05) with 14% walnuts. In the metabolite-profiling analysis, walnuts caused a significant increase in several polyunsaturated fatty acids (PUFAs), including docosahexaenoic acid (DHA) and 9-oxo-10(E),12(E)-octadecadienoic acid (9-oxoODA), as well as kynurenic acid. In colon tissue samples, walnuts caused a significant increase in the levels of S-adenosylhomocysteine (SAH) and betaine, important components of fatty acid β-oxidation. These metabolite changes may contribute in part to the observed protection against DSS-induced inflammatory tissue injury.
McArthur, B., R. Mattes, 2019. Energy extraction from nuts: walnuts, almonds, pistachios. Br J Nutr. 17:1-21.
The bioaccessibility of fat has implications for satiety and postprandial lipidemia. The prevailing view holds that the integrity of plant cell wall structure is the primary determinant of energy and nutrient extraction from plant cells as they pass through the gastrointestinal tract. However, comparisons across nuts (walnuts, almonds, pistachios) with varying physical properties do not support this view. In this study, masticated samples of three nuts from healthy adults were exposed to a static model of gastric digestion followed by simulated intestinal digestion. Primary outcomes were particle size and lipid release at each phase of digestion. Walnuts produced a significantly larger particle size post-mastication compared to almonds. Under gastric and intestinal conditions, the particle size was larger for walnuts compared to pistachios and almonds (P<0.05). However, the masticated and digesta particle sizes were not related to the integrity of cell walls nor lipid release. The total lipid release was comparable between nuts after the in vitro intestinal phase (P>0.05). Microstructural examination showed ruptured and fissured cell walls that would allow digestion of cellular contents and this may be governed by internal cellular properties such as oil body state. Furthermore, the cell walls of walnuts tend to rupture rather than separate and as walnut tissue passes through the gastrointestinal track, lipids tend to coalesce reducing digestion efficiency.
Koh, S.J., Y.I. Choi, Y. Kim, Y.S. Kim, S.W. Choi, J.W. Kim, B.G. Kim, K.L. Lee, 2019. Walnut phenolic extract inhibits nuclear factor kappaB signaling in intestinal epithelial cells, and ameliorates experimental colitis and colitis-associated colon cancer in mice.
Eur J Nutr. 58(4):1603-1613.
Purpose: Walnuts (Juglans regia) are known to have anti-cancer and immunomodulatory effects. However, little information is available on the effects of walnut phenolic extract (WPE) on intestinal inflammation and colitis-associated colon cancer. Methods: COLO205 cells were pretreated with WPE and then stimulated with tumor necrosis factor (TNF)-α. In the acute colitis model, wild type mice (C57BL/6) were administered 4% dextran sulfate sodium (DSS) for 5 days. In the chronic colitis model, interleukin (IL)-10-/- mice were administered with either the vehicle or WPE (20 mg/kg) by oral gavage daily for 2 weeks. In an inflammation-associated tumor model, wild type mice were administered a single intraperitoneal injection of azoxymethane followed by three cycles of 2% DSS for 5 days and 2 weeks of free water consumption. Results: WPE significantly inhibited IL-8 and IL-1α expression in COLO205 cells. WPE attenuated both the TNF-α-induced IκB phosphorylation/degradation and NF-κB DNA binding activity. The administration of oral WPE significantly reduced the severity of colitis in both acute and chronic colitis models, including the IL-10-/- mice. In immunohistochemical staining, WPE attenuated NF-κB signaling in the colons of both colitis models. Finally, WPE also significantly reduced tumor development in a murine model of colitis-associated colon cancer (CAC). Conclusions: WPE ameliorates acute and chronic colitis and CAC in mice, suggesting that WPE may have potentials for the treatment of inflammatory bowel disease.
Hwang, H.J., Y. Liu, H.S. Kim, H. Lee, Y. Lim, H. Park, 2019. Daily walnut intake improves metabolic syndrome status and increases circulating adiponectin levels: randomized controlled crossover trial. Nutr Res Pract. 13(2):105–114.
Background/Objectives: Several previous studies have investigated whether regular walnut consumption positively changes heart-health-related parameters. The aim of this study was to investigate the effects of daily walnut intake on metabolic syndrome (MetS) status and other metabolic parameters among subjects with MetS. Materials/Methods: This study was a two-arm, randomized, controlled crossover study with 16 weeks of each intervention (45 g of walnuts or iso-caloric white bread) with a 6-week washout period between interventions. Korean adults with MetS (n = 119) were randomly assigned to one of two sequences; 84 subjects completed the trial. At each clinic visit (at 0, 16, 22, and 38 weeks), MetS components, metabolic parameters including lipid profile, hemoglobin A1c (HbA1c), adiponectin, leptin, and apolipoprotein B, as well as anthropometric and bioimpedance data were obtained. Results: Daily walnut consumption for 16 weeks improved MetS status, resulting in 28.6%-52.8% reversion rates for individual MetS components and 51.2% of participants with MetS at baseline reverted to a normal status after the walnut intervention. Significant improvements after walnut intake, compared to control intervention, in high-density lipoprotein cholesterol (HDL-C) (P = 0.028), fasting glucose (P = 0.013), HbA1c (P = 0.021), and adiponectin (P = 0.019) were observed after adjustment for gender, age, body mass index, and sequence using a linear mixed model. Conclusion: A dietary supplement of 45 g of walnuts for 16 weeks favorably changed MetS status by increasing the concentration of HDL-C and decreasing fasting glucose level. Furthermore, consuming walnuts on a daily basis changed HbA1c and circulating adiponectin levels among the subjects with MetS.
