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Acknowledgements | Open Access funding was provided by the University of the Basque Country. | PMC9469834 |
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Author contribution | Arteagoitia Iciar conceived the idea, designed the protocol, and led the acquisition of funds; Barbier Luis conceived the idea, led the acquisition of funds, and reviewed the protocol. Santamaria conducted the trial and collected the data; Rodriguez-Sanchez and Rodriguez-Andrés collected and analysed the data. All authors have reviewed and approved the final manuscript. | PMC9469834 |
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Funding | Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Open Access funding provided by University of the Basque Country. This clinical trial has been funded by the Instituto de Salud Carlos III through the project “PI18/00809″ (Co-funded by European Regional Development Fund/European Social Fund; “A way to make Europe”/”Investing in your future”). | PMC9469834 |
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Data availability | The data that support the findings of this study are available from the corresponding author upon reasonable request. | PMC9469834 |
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Declarations | PMC9469834 |
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Ethics approval | All procedures performed in the studies involving human participants complied with the ethical standards of the institutional and/or national research committee, and with the 1964 Helsinki declaration, and its later amendments or comparable ethical standards. This study was approved by The Spanish Agency of Medicines and Medical Devices (AEMPS) and the Euskadi Research Ethics Committee under the protocol number N° 2018062. | PMC9469834 |
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Conflict of interest | The authors declare no competing interests. | PMC9469834 |
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References
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1. Introduction | SECONDARY | Background: The Dietary Guidelines for Americans (DGA) recommends consuming a variety of “Protein Foods” based on “ounce-equivalent” (oz-eq) portions. No study has assessed the same oz-eq portions of animal- vs. plant-based protein foods on essential amino acid (EAA) bioavailability for protein anabolism in young and older adults. Objectives: We assessed the effects of consuming two oz-eq portions of pork, eggs, black beans, and almonds on postprandial EAA bioavailability in young and older adults. Methods: We conducted two investigator-blinded, randomized crossover trials in young (The 2020–2025 Dietary Guidelines for Americans (DGA) recommend that “Protein Foods” should be consumed as part of a healthy dietary pattern [Research by Park et al. [Therefore, we conducted two sequentially randomized, investigator-blinded, crossover acute feeding trials with the same study design: the first in a cohort of young adults and the second in a cohort of older adults. The primary objective of this project was to assess the effect of consuming two oz-eq portions of animal-based (unprocessed lean pork or whole eggs) vs. plant-based (black beans or sliced almonds) protein foods as part of a mixed whole foods meal on plasma EAA bioavailability for protein anabolism. We hypothesized that consuming a meal with two oz-eq portions of animal-based protein foods would result in greater postprandial EAA bioavailability compared to plant-based protein foods (primary) and would not elicit differential EAA bioavailability responses between young and older adults (secondary). This research will serve as an important resource for future DGAs to reevaluate the appropriateness of equating different protein sources comprising the protein foods group on an oz-eq basis for young and older adults. | PMC10343739 |
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2. Materials and Methods | PMC10343739 |
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2.1. Ethics | RECRUITMENT | The study protocols and materials were approved by the Purdue University Biomedical Institutional Review Board (IRB protocol #1804020520 (young adults) and IRB-2019-354 (older adults)), and all participants provided written, informed consent, and received monetary compensation for their participation. Before participant recruitment, these studies were registered at clinicaltrials.gov as NCT03649568 (young adults) and NCT04243395 (older adults). The reporting of this research followed the CONSORT reporting guidelines [ | PMC10343739 |
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2.2. Participants | Participants were recruited from Greater Lafayette, IN, USA. Study inclusion criteria were: aged 22–39 or 55–75 years for young and older adults, respectively; BMI 20–35 kg/m | PMC10343739 |
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2.3. Study Design | Each participant completed four 300-min trials (randomized, investigator-blinded, crossover design), with each trial separated by a minimum of three days. Participant treatment order randomization was performed by a clinical laboratory manager, who did not participate in data analysis or interpretation, using online software (Randomization.com). While the participants, clinical laboratory manager, and dietitians were not blinded, the investigators were blinded until all participants finished the protocol and all sample analyses were completed. Before each testing day, participants were asked to refrain from performing physical activity for 48 h. Participants were provided a controlled meal to consume to satiation the evening before each testing day (1052 kcal: fat: 25.5 g; carbohydrate: 163.6 g; protein: 47.2 g) ( | PMC10343739 |
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2.4. Test Meals | On each of the four testing days, participants consumed a carefully portioned test meal ( | PMC10343739 |
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2.5. Sample Collection and Biochemical Analyses | BLOOD | Blood samples were collected into serum- and plasma-separator tubes at the specified time points, centrifuged for 15 min at 4000 rpm and 4 °C, aliquoted into 1 mL microcentrifuge tubes, and stored at −80 °C as previously described [ | PMC10343739 |
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2.6. Statistical Analysis | Previous research from our group with a similar design and with EAA positive incremental area under the curve (iAUCpos) as the primary outcome showed a between-trial, within-subject variability of ±2000 μg/mL/h [ | PMC10343739 |
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3. Results | PMC10343739 |
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3.1. Randomization and Participant Characteristics | Details of the study enrollment and conduct for this project are described in | PMC10343739 |
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3.1.1. Young Adults | Of the 39 potential participants who completed an initial in-person screening, 37 were enrolled and randomized, and 30 (15 females and 15 males) completed the study. Seven participants withdrew from the study: two due to starting a new job and relocating; two due to personal reasons; two chose to discontinue after the first trial; and one due to challenges placing intravenous catheter lines. The mean (±SD) age of participants was 26.0 (±4.9) years, BMI was 26.4 (±4.5) kg/m | PMC10343739 |
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3.1.2. Older Adults | ADVERSE EVENTS | Of the 30 potential participants who completed an initial in-person screening, 27 were enrolled and randomized, and 25 (15 females and 10 males) completed the study. Two participants chose to discontinue after the first trial because of the time commitment. The mean (±SD) age of participants was 64.2 (±6.6) years, BMI was 26.1 (±3.7) kg/mFor this project, there were no adverse events reportable to the Purdue University Institutional Review Board. | PMC10343739 |
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3.2. Baseline Fasting Blood Concentrations | TAA | Among the four trials, baseline fasting TAA, EAA, BCAA, leucine, glucose, and insulin were not different for young adults, older adults, or combined. Among the four trials, baseline fasting EAA, BCAA, and leucine were greater for young than older adults, with no age-related differences in TAA, glucose, or insulin. | PMC10343739 |
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3.3. Essential Amino Acids | PMC10343739 |
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3.3.1. Young Adults | pork | A main effect of protein oz-eq food sources was observed for EAA iAUCpos. Essential amino acid iAUCpos were greater for pork or eggs than for black beans or almonds; greater for pork than eggs; and not different between black beans and almonds ( | PMC10343739 |
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3.3.2. Older Adults | pork | A main effect of protein oz-eq food sources was observed for EAA iAUCpos. Essential amino acid iAUCpos was greater for pork or eggs than for black beans or almonds; greater for pork than for eggs; and not different between black beans and almonds ( | PMC10343739 |
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3.3.3. Young vs. Older Adults | Essential amino acid iAUCpos was not different between young and older adults overall or among trials ( | PMC10343739 |
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3.3.4. Young and Older Adults Combined | pork | A main effect of protein oz-eq food sources was observed for EAA iAUCpos. Essential amino acid iAUCpos was greater for pork or eggs than black beans or almonds, greater for pork than eggs, and not different between black beans and almonds ( | PMC10343739 |
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3.4. TAA, BCAA, and Leucine | TAA | The responses and statistical results (i.e., comparisons among trials and each time point) for TAA, BCAA, and leucine were comparable to EAA for young adults, older adults, young vs. older adults, and combined ( | PMC10343739 |
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3.5. Serum Glucose and Insulin | PMC10343739 |
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3.5.1. Young Adults | No differences were observed for glucose or insulin iAUCpos overall or among trials ( | PMC10343739 |
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3.5.2. Older Adults | No differences were observed for glucose iAUCpos overall or among trials (Insulin iAUC trended toward being greater for black beans than almonds ( | PMC10343739 |
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3.5.3. Young vs. Older Adults | A main effect of age was observed for glucose iAUCpos. Glucose iAUCpos was lower for young than older adults for black beans but not for pork, eggs, or almonds (Insulin iAUCpos was not different between young and older adults overall or among trials ( | PMC10343739 |
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3.5.4. Young and Older Adults Combined | There was no main effect of protein oz-eq food source on glucose iAUCpos, although it trended toward significance (A main effect of protein oz-eq food sources was observed for insulin. Insulin iAUCpos was not different among trials, although it trended toward being greater for black beans than for almonds ( | PMC10343739 |
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3.6. Females vs. Males | TAA | The TAA, EAA, BCAA, leucine, glucose, and insulin concentrations for each time point and iAUCpos are reported in | PMC10343739 |
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3.6.1. Baseline Fasting Blood Concentrations | TAA | Baseline fasting EAA, BCAA, and leucine were lower for females than males, with no sex-specific differences in TAA, glucose, or insulin. | PMC10343739 |
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3.6.2. Plasma Amino Acids | A main effect of sex and treatment × sex was observed for EAA iAUCpos. Essential amino acid iAUCpos were greater for females than males overall and for pork, but not for eggs, black beans, or almonds ( | PMC10343739 |
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3.6.3. Serum Glucose and Insulin | Glucose and insulin iAUCpos were not different between females and males overall or among trials ( | PMC10343739 |
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4. Discussion | SECONDARY | To the best of the author’s knowledge, this is the first project to assess the effect of consuming the same oz-eq portions of protein foods, namely pork, eggs, black beans, and almonds, as defined by the DGA, in the context of a mixed meal on EAA substrate bioavailability for protein anabolism in young and older adults. Consistent with our hypotheses, consuming a meal with two oz-eq of unprocessed lean pork or whole eggs resulted in greater postprandial EAA bioavailability compared to a meal with two oz-eq of black beans or raw sliced almonds in (1) young adults; (2) older adults; and (3) young and older adults combined. No differences were observed between young and older adults. In addition, lean pork resulted in greater EAA bioavailability than eggs in young adults and older adults, separately and combined. These findings show that, on the same oz-eq basis, consuming these animal- vs. plant-based protein foods more effectively provides bioavailable EAA for protein anabolism.These findings are consistent with research by Park et al. [Taken together, the findings from our project and others [Consistent with our secondary hypothesis, we did not observe differential EAA iAUCpos responses between young and older adults overall or among protein foods. There is consistent evidence that basal (fasting) protein synthesis does not differ between young and older adults [Our observation that postprandial EAA iAUCpos were greater for females than for males for pork may be due to the females consuming more protein relative to body mass and plasma volume. However, limited evidence suggests the anabolic response to the same protein/AA ingestion or infusion is not different between young [Regarding project strengths and limitations, this is the first project to assess the effects of consuming the same oz-eq portions of protein foods as defined by the DGA on postprandial EAA responses in the context of mixed whole food meals in both young and older adults. We chose to test a two oz-eq portion of each Protein Food to theoretically obtain measurable postprandial EAA responses. We are mindful that this specific portion likely does not reflect the amounts of these protein foods consumed on a meal-to-meal or weekly basis by young and older adults. Importantly, with consideration of protein quantity, EAA content and bioavailability, and disparate metabolizable energy contents, these results highlight the shortcomings of using oz-eq to achieve the DGA recommendations for protein foods. Our combined sample size of 55 (30 females, 25 males) participants exceeded our a priori estimated need based on effect size, and we used conservative Bonferroni-adjusted | PMC10343739 |
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5. Conclusions | In conclusion, based on the oz-eq concept used in the DGAs, the animal- and plant-based protein foods included in this project do not equivalently provide bioavailable EAA for protein anabolism in young and older adults. Whole eggs and especially unprocessed lean pork consumed with a standard meal resulted in greater postprandial EAA bioavailability than black beans or raw sliced almonds. This research demonstrates that the word “equivalent” in the oz-eq unit of measure for protein foods does not apply to the protein content and postprandial essential amino acid bioavailability of these foods. | PMC10343739 |
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Supplementary Materials | The following supporting information can be downloaded at: Click here for additional data file. | PMC10343739 |
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Author Contributions | A.S.H. | Conceptualization, J.L.H., R.E.B. and W.W.C.; Data curation, G.C., E.M.D., A.S.H. and W.Z.; Formal analysis, G.C., A.S.H. and W.Z.; Funding acquisition, J.L.H., R.E.B. and W.W.C.; Investigation, G.C. and E.M.D.; Methodology, G.C., J.L.H., R.E.B., C.C.C. and W.W.C.; Project administration, G.C. and W.W.C.; Resources, C.C.C. and W.W.C.; Supervision, W.W.C.; Writing—original draft, G.C.; Writing—review and editing, G.C., J.L.H., R.E.B., E.M.D., A.S.H., W.Z., C.C.C. and W.W.C.; W.W.C. had the primary responsibility for the final content. All authors have read and agreed to the published version of the manuscript. | PMC10343739 |
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Institutional Review Board Statement | This project was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Board of Purdue University (IRB protocol #1804020520 (young adults) and IRB-2019-354 (older adults)). | PMC10343739 |
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Informed Consent Statement | Informed consent was obtained from all subjects involved in the study. | PMC10343739 |
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Data Availability Statement | The data supporting this project’s findings are available from the corresponding author, W.W.C., upon reasonable request. | PMC10343739 |
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Conflicts of Interest | When this research was conducted, W.W.C. received research funding from the following organizations: American Egg Board’s Egg Nutrition Center, Beef Checkoff, Pork Checkoff, North Dakota Beef Commission, Barilla Group, Mushroom Council, and the National Chicken Council. C.C.C. received funding from the Beef Checkoff. R.E.B. is currently employed by Archer-Daniels-Midland (ADM); the research presented in this article was conducted in a former role and has no connection with ADM. G.C., J.L.H., E.M.D., A.S.H. and W.Z. declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. | PMC10343739 |
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1. Introduction | non-alcoholic fatty liver disease, cardiometabolic abnormalities, Hypertension, hypertension, T2DM | OBESE, NON-ALCOHOLIC FATTY LIVER DISEASE, HYPERGLYCEMIA, METABOLIC DISEASES, INSULIN RESISTANCE, ADIPOSITY, HYPERTENSION, METABOLIC SYNDROME, HYPERTENSION, DYSLIPIDEMIA | This feeding trial evaluated the impact of the Dietary Approaches to Stop Hypertension diet on changes in plasma choline, choline metabolites, and ceramides in obese older adults; 28 adults consumed 3oz (n = 15) or 6oz (n = 13) of beef within a standardized DASH diet for 12 weeks. Plasma choline, betaine, methionine, dimethylglycine (DMG), phosphatidylcholine (PC), lysophosphotidylcholine (LPC), sphingomyelin, trimethylamine-N-oxide (TMAO), L-carnitine, ceramide, and triglycerides were measured in fasted blood samples. Plasma LPC, sphingomyelin, and ceramide species were also quantified. In response to the study diet, with beef intake groups combined, plasma choline decreased by 9.6% (Aging presents with body composition changes that include declines in muscle health with simultaneous increases in body fat. These changes increase the risk for cardiometabolic abnormalities such as insulin resistance, hyperglycemia, visceral adiposity, dyslipidemia, and hypertension [Choline, a micronutrient grouped with the family of B-complex vitamins, has received considerable attention due to its role in one-carbon metabolism, neurotransmission, membrane synthesis, and lipid transport [Ceramide is a sphingolipid that can be synthesized from the hydrolysis of sphingomyelin [The Dietary Approaches to Stop Hypertension (DASH) dietary pattern is a high-quality therapeutic diet known to improve health in vulnerable populations, resulting in improved cardiovascular and cognitive health and reductions in metabolic diseases, including T2DM, metabolic syndrome, and non-alcoholic fatty liver disease [ | PMC10489641 |
2. Materials and Methods | PMC10489641 |
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2.1. Study Participants | RECRUITMENT | Characteristics and recruitment of participants have been previously reported [ | PMC10489641 |
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2.2. Study Design, Body Composition Measures, and Blood Sample Collection | CLOT, BLOOD, HEART, LUNG | A human controlled feeding trial with a parallel design was performed in which females (n = 17) and males (n = 11) aged 65+ years were randomized to consume either 3 oz (85 g; n = 15) or 6oz (170.1 g; n = 13) of lean fresh beef within a standardized DASH diet. As previously described, Nutritionist Pro™ software was used to create the study diet. The nutrient composition was based upon the 2015–2020 dietary guidelines for daily caloric intake for older sedentary adults and the DASH eating plan using the National Heart, Lung, and Blood Institute, National Institutes of Health [Body composition and cardiometabolic methods have been previously reported [Participants provided fasted blood samples at weeks 0, 3, 6, 9, and 12. Due to limited resources for analysis, for this study measures were performed at weeks 0, 6, and 12. The fasted blood samples were collected into 10 mL serum separator clot activator tubes (SST Vacutainer; Pulmolab, Northridge, CA, USA) and EDTA-coated tubes (Pulmolab) by a trained phlebotomist. Blood for serum collection was kept at room temperature, allowed to clot, and centrifuged at 650× | PMC10489641 |
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2.3. Choline, Betaine, Trimethylamine N-Oxide, and L-Carnitine Quantification | We utilized LC-MS/MS to quantify choline and betaine as previously described [ | PMC10489641 |
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2.4. Ceramide Quantification | dryness | Plasma samples (700 µL) were frozen over liquid nitrogen and lyophilized to dryness. HPLC-grade methanol (1.4 mL) was added to the dried material and the mixture was sonicated for 3 min (on/off pulse cycles of 3 s on, 2 s off, at power 100%) using a Qsonica Ultrasonic Processor (Model Q700; Qsonica, Newtown, CT, USA) with a water bath cup horn adaptor (Model 431C2) and the water bath maintained at room temperature. Samples were placed on an end-over-end rotator, and extraction continued for 24 h, then centrifuged at 18,000× Ceramide analyses were performed using a Thermo Scientific Vanquish Horizon UHPLC System coupled with a Thermo Scientific TSQ Quantis Triple Quadrupole mass spectrometer equipped with a HESI ion source. Mobile phase A was 78.6% water, 20% acetonitrile, and 0.4% formic acid ( | PMC10489641 |
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2.5. Lysophosphatidylcholine Quantification | SEPARATION, RESOLUTION | Stock lipid standards were prepared by dissolving them in dichloromethane (DCM)/methanol (MeOH; 2:1 Thirty microliters of internal standard mixture were added to 30 μL of sample followed by 190 μL of MeOH. Samples were then vortexed for 20 s. Next, 380 μL of DCM was added, the sample was vortexed for 20 s, and 120 μL of water was added to induce phase separation. The samples were then vortexed for 10 s and allowed to equilibrate at room temperature for 10 min before centrifugation at 8000× Chromatographic separation was performed on a Vanquish UHPLC system with an Accucore C30, 2.6 μm column (2.1 mm id × 150 mm) coupled to a Q Exactive™ Hybrid Quadrupole-Orbitrap High Resolution Mass Spectrometer (Thermo Scientific). The mobile phase consisted of (solution A) 60% ACN, 40% water, 10 mM ammonium formate with 0.1% formic acid and (solution B) 90% IPA, 10% ACN, 10 mM ammonium formate with 0.1% formic acid. The gradient was as follows: 0 to 1.5 min, 32% solvent B; 1.5 to 4 min, 32 to 45% solvent B; 4 to 5 min, 45 to 52% solvent B; 5 to 8 min 52 to 58% solvent B; 8 to 11 min, 58 to 66% solvent B; 11 to 14 min, 66 to 70% solvent B; 14 to 18 min, 70 to 75% solvent B; 18 to 21 min, 75 to 97% solvent B; 21 to 25 min, isocratic 97% solvent B; 25 to 25.1 min, 97 to 32% solvent B; followed by 4 min of re-equilibration of the column before the next run. The flow rate was 260 μL/min. All of the samples were analyzed by electrospray ionization in data-dependent MS/MS mode.Nitrogen was used as the nebulizing gas, with sheath, auxiliary, and sweep gas flows set at 50, 5, and 1 arbitrary units, respectively. Other conditions included the following: resolution, 120,000 full width at half maximum; automatic gain control target, 3 × 10Acquired data were processed using LipidSearch™ software version 4.3 (Thermo Scientific) with the following workflow: first, the individual data files were searched for product ion MS/MS spectra of lipid precursor ions. MS/MS fragment ions were predicted for all precursor adduct ions measured within ±5 ppm. The product ions that matched the predicted fragment ions within a ±5 ppm mass tolerance were used to calculate a match-score, and those candidates providing the highest-quality match were determined. Next, the search results from the individual positive or negative ion files from each sample group were aligned within a retention time window (±0.1 min) and the data were merged for each annotated lipid. The annotated lipids were then filtered to reduce false positives. | PMC10489641 |
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2.6. Statistical Analyses | Data were tested for assumptions of normality and spread using the Shapiro–Wilk test and normal distribution curves. Data that did not meet assumptions of normality were analyzed with non-parametric tests and reported as the median with interquartile range. Data were pooled by sex and by beef intake group and also displayed separately. Independent samples | PMC10489641 |
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3. Results | PMC10489641 |
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3.1. Study Diet | Estimated choline, choline forms, L-carnitine, and methionine content of the study diet separated by beef group are shown in Given that the 2022 USDA database does not include choline forms, glycerophosphocholine (GPC), phosphocholine, PC, and sphingomyelin were calculated using the 2008 USDA Database for the Choline Content of Common Foods [L-carnitine is not included in the USDA database, thus the National Institutes of Health Office of Dietary Supplements Carnitine Fact Sheet for Health Professionals was used to calculate dietary L-carnitine in the beef provided by the study diet [Nutritionist Pro™ software was used to estimate dietary methionine content of the diet. Each food item from the study was put into the software and Nutritionist Pro™ calculated the amount of methionine consumed. Participants in the 3 oz group received 1350 mg of methionine and the 6 oz group received 1957 mg of methionine. | PMC10489641 |
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3.2. Baseline (Week 0) Characteristics of Study Participants | Participants (n = 28) 65–84 years of age completed the 12-week diet intervention and were included in the final analysis. As previously reported, all participants presented with the following at baseline: total body weight: 91.2 kg; BMI: 32 kg/mBaseline characteristics separated by beef group are presented in Baseline characteristics separated by sex are presented in | PMC10489641 |
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3.3. Week 12 Outcomes in Response to the Intervention | Week 12 results separated by beef group are presented in Week 12 outcomes separated by sex are presented in | PMC10489641 |
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3.4. Changes in Plasma Choline, Choline Metabolites, Total Ceramide, and Triglycerides in Response to the Study Diet | Plasma choline, betaine, methionine, DMG, PC, total LPC, total sphingomyelin, TMAO, L-carnitine, total ceramide, and triglyceride responses to the diet intervention are shown in | PMC10489641 |
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3.5. Changes in LPC, Sphingomyelin, and Ceramide Species in Response to the Diet Intervention | Baseline characteristics of LPC, sphingomyelin, and ceramide species separated by beef group are presented in Week 12 results of LPC, sphingomyelin, and ceramide species separated by beef group are presented in Changes in plasma LPC, sphingomyelin, and ceramide species in response to the diet intervention are presented in | PMC10489641 |
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3.6. Changes in Choline, Betaine, L-Carnitine, TMAO, Sphingomyelins, Ceramides, and LPCs in Correlation with Changes in Body Composition, Cardiometabolic, and Inflammatory Markers | inflammation | INFLAMMATION | Changes in choline, betaine, LPCs, sphingomyelins, TMAO, L-carnitine, and ceramides in association with body composition, cardiometabolic, and inflammatory markers are shown in The change in sphingomeyelin 24:0 was correlated with total cholesterol (r = −0.42; Several LPC species were correlated with anthropometric and biomarkers of cardiometabolic health and inflammation (Numerous LPC species were correlated with the inflammatory markers CRP and IL-8. The change in LPC 16:1, 18:0, 18:1 18:1e, 18:2, 18:3, 20:0 20:1, 20:3, 20:4, and 22:5 was correlated with CRP ( | PMC10489641 |
4. Discussion | OBESE | This highly controlled diet intervention study sought to examine the impact of a DASH-style diet on changes in choline, choline metabolites, ceramides, and triglycerides in a cohort of obese adults aged 65–84 years. In response to the 12-week intervention, changes in plasma choline, DMG, total PC, total LPC, TMAO, total ceramide, and triglycerides resulted. Furthermore, the study diet influenced changes in individual LPC, sphingomyelin, and ceramide species. | PMC10489641 |
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4.1. In Response to the Dietary Choline and Betaine Provided by the DASH Diet, Plasma Choline Decreased and in Males Plasma Betaine Increased | The study diet provided an average of ~300 mg of dietary choline per day, which was delivered by food only, supplemental choline was not given. The amount of choline supplied by the study diet was below the adequate intake (AI) for adults which is 550 mg per day for males and 425 mg for females [In response to the study diet, plasma choline decreased by 9.6% from baseline to study end (Estimated dietary betaine provided by the study diet had an average of 31.5 mg provided by food only, supplemental betaine was not given. The amount of betaine in the study diet was below dietary intakes reported by other studies. A 2017 systematic review and meta-analysis of six prospective cohort studies using FFQs reported a dietary betaine intake range of 41–478 mg in adults [In the present study, males had higher plasma betaine compared to females at week 12 ( | PMC10489641 |
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4.2. Plasma Dimethylglycine Decreased in Response to the DASH Diet, and Males Had Higher DMG Compared to Females | In the present study, the average estimated amount of dietary methionine was 1654 mg. In response to the study diet, plasma DMG decreased by 10% (At baseline, males had higher plasma DMG ( | PMC10489641 |
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4.3. Plasma Phosphatidylcholine Decreased in Response to the Dietary Phosphatidylcholine Provided by the DASH Diet | obese, overweight, prediabetes and/or metabolic syndrome | OBESE | An estimated average of 81.5 mg of dietary phosphatidylcholine (PC) was provided by the study diet. The primary food sources of dietary PC were beef and eggs and in the present study beef was the main PC food item. Conflicting outcomes on metabolic health related to beef or dietary PC have been reported. A 28-day randomized crossover controlled feeding trial in overweight and obese adults aged 18–74 years with prediabetes and/or metabolic syndrome showed that beef within the Healthy US-Style Eating Pattern had no effects on cardiometabolic health [In response to the 12-week diet intervention, plasma total PC decreased by 51% ( | PMC10489641 |
4.4. In Response to the DASH Diet Total Lysophosphatidylcholine Increased | The physiological functions and specific mechanisms by which lysophosphatidylcholine (LPC) acts have not been fully elucidated and human clinical lipidomic trials report conflicting outcomes [ | PMC10489641 |
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4.5. Trimethylamine N-Oxide Increased in Response to a Higher Beef Intake | Trimethylamine | PMC10489641 |
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4.6. Total Ceramide Decreased in Response to the DASH Diet | T2DM | INSULIN RESISTANCE, OBESE | The bioactive sphingolipid ceramide receives attention due to its effects on insulin signaling and glucose utilization. Indeed, circulating ceramide concentrations are elevated in obese adults with T2DM and correlate with the severity of insulin resistance [ | PMC10489641 |
4.7. In Response to the DASH Diet Plasma Triglycerides Decreased and Males Had Higher Triglycerides Compared to Females | metabolic syndrome, middle-aged overweight | METABOLIC SYNDROME | Studies show that the DASH diet significantly reduces plasma triglycerides in middle-aged overweight adults and individuals with metabolic syndrome [At baseline, participants randomized to the 3 oz beef group had higher ( | PMC10489641 |
4.8. Lysophosphatidylcholine Species Increased in Association with Biomarkers of Inflammatory and Muscle Health | inflammation, atherosclerosis | INFLAMMATION, ATHEROSCLEROSIS | Lysophosphatidylcholines (LPCs) are bioactive lipids investigated in the development of atherosclerosis and inflammation [LPCs are widely considered to be potent pro-inflammatory lipids, however, recent discoveries suggest that LPCs may possess anti-inflammatory properties as well [ | PMC10489641 |
4.9. Sphingomyelin Species Respond Differentially in Association with Body Composition and Cardiometabolic Outcomes | metabolic diseases | METABOLIC DISEASES | Estimated dietary sphingomyelin provided by the study had an average of 17.5 mg. Dietary sphingomyelins are shown to be beneficial in lipid metabolism, cholesterol regulation, and in the prevention and treatment of metabolic diseases [At baseline, females had higher plasma sphingomyelin compared to males ( | PMC10489641 |
4.10. In Response to the DASH Diet Ceramide Species Respond Differentially | The impact of diet on the profile of individual ceramide species is unexplored. In the present study, plasma concentrations of five ceramide species were measured. Although in response to the study diet total ceramide decreased ( | PMC10489641 |
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4.11. Limitations | Limitations of the present study include: (i) although two different amounts of beef were compared, this study lacked a non-beef comparison group; (ii) all participants self-identified as white American which is representative of the main racial group in South Dakota; (iii) no participants required support for daily living activities and lived independently in their own homes; (iv) South Dakota is a rural state. These limitations should be considered when generalizing outcomes from this study to diverse populations of older adults including those with differing demographic and ethnic backgrounds and living environments. | PMC10489641 |
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5. Conclusions | OBESE | Results from this controlled feeding diet intervention trial in obese older adults show that in response to the DASH diet that provided dietary choline below the AI, plasma choline, PC, and total ceramide decreased, while plasma total LPC and TMAO increased. Outcomes also demonstrated distinct patterns of change in plasma LPC, sphingomyelin, and ceramide species, suggesting that the DASH diet may influence the determination and modification of individual lipid species. Given that the effect of diet on changes in specific phospholipid and sphingolipid molecules in relation to metabolic health outcomes in humans remains largely unexplored, additional studies are required to determine the long-term clinical effects and relevance of these relationships, particularly in vulnerable populations such as older adults. | PMC10489641 |
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Supplementary Materials | The following supporting information can be downloaded at: Click here for additional data file. | PMC10489641 |
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Author Contributions | Conceptualization, C.A.P., B.N.T. and J.W.M.; methodology, C.A.P. and J.W.M.; statistical analysis, G.P.V.G.; data curation, B.N.T., M.A., L.A.S., M.E.D.-R., H.H.L. and E.L.J.; writing—original draft preparation, C.A.P., B.N.T., G.P.V.G. and M.A.; writing—review and editing, C.A.P., G.P.V.G., M.A., L.A.S., B.N.T. and J.W.M.; supervision, C.A.P. and J.W.M.; project administration, C.A.P. and J.W.M.; funding acquisition, C.A.P. All authors have read and agreed to the published version of the manuscript. | PMC10489641 |
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Institutional Review Board Statement | The study was conducted in accordance with the Declaration of Helsinki. The protocol was reviewed and approved by the Institutional Review Board for Human Study Participant Use at South Dakota State University (Approval #: IRB-1,712,006-EXP). | PMC10489641 |
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Informed Consent Statement | Informed consent was obtained from all participants involved in this study prior to enrollment. | PMC10489641 |
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Data Availability Statement | Data presented are contained within this article and | PMC10489641 |
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Conflicts of Interest | The authors declare no conflict of interest. | PMC10489641 |
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Abbreviations | PMC10489641 |
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References | Estimated dietary choline, betaine, choline forms, L-carnitine, and methionine provided by the study diet.* Estimated using 2022 USDA FoodCentral database [Baseline and week 12 variable characteristics separated by beef group.* Baseline data are presented as means and standard deviations except for TMAO and L-carnitine which are reported as median (interquartile range). Independent samples Baseline and week 12 variable characteristics separated by sex.* Baseline data are presented as means and standard deviations except for TMAO and L-carnitine which are reported as median (interquartile range). Independent samples Choline, choline metabolites, ceramide, and triglycerides in response to the 12-week diet intervention.Data are presented as means and standard deviations, except for betaine, methionine, DMG, and L-carnitine which are reported as median (interquartile range). For normally distributed data, a mixed design ANOVA was used to determine changes in the primary outcome variables across the intervention. For betaine, methionine, DMG, and L-carnitine, the Friedman’s test was used to determine changes from baseline to week 12. DMG, dimethylglycine; PC, phosphatidylcholine; LPC, lysophosphatidylcholine; TMAO, trimethylamine N-oxide. † Changes in LPC, sphingomyelin, and ceramide species in response to the diet intervention.Data are presented as means and standard deviations, except for LPC (weeks of intervention) and ceramide species which are reported as median (interquartile range). For normally distributed data, independent samples | PMC10489641 |
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Background | POSITIVE | Positive psychology interventions are known to have an impact on mental health as well as on a number of beneficial characteristics like optimism, gratitude and self-efficacy. The Positive Emotions Training (PoET) is one of the first holistic training programs covering eleven positive psychology constructs. The goal of this study was to test PoET’s feasibility in the general population and to assess possible effects on positive and negative mental health factors. Additionally, possible effects on optimism, gratitude, happiness, resilience, and self-efficacy were examined.
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Methods | The sample ( | PMC10426081 |
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Results | depression, anxiety | The results showed that the contents were comprehensible and that the conduction of the training was feasible overall. In addition, a significant decrease of depression and anxiety symptoms as well as a significant increase of optimism were found in the PoET group. No significant changes were found in the control group.
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Conclusions | Results indicate that PoET is an applicable intervention for improving mental health in the general population. | PMC10426081 |
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Trial registration | Retrospectively registered at ClinicalTrials.gov on 21/02/2023 (Identifier/Trial registration number: NCT05737251). | PMC10426081 |
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Keywords | Open Access funding enabled and organized by Projekt DEAL. | PMC10426081 |
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Background | depression, anxiety | POSITIVE, DISORDERS | Mental disorders affect millions of people worldwide, causing suffering and various negative consequences. According to the World Health Organization (WHO), the lifetime prevalence ranges from 18.1 to 36.1% [Several studies highlight the psychological burden of the COVID-19 pandemic and related restrictions [For a long time, psychology focused on curing existing psychopathologies. Positive psychology has shifted the focus from dealing only with the negative aspects of life to further improving positive qualities [However, most studies so far have examined individual exercises, but rarely entire holistic training programs. Mostly, such training programs were designed for specific target groups. The first training program of this kind was developed to increase happiness and life satisfaction in college students. It extended over six weeks and was able to significantly increase happiness of the participants [Overall, the literature supports the effectiveness of positive psychology training programs. However, as described above, research so far has focused on testing the effectiveness of such trainings in specific target groups or individual exercises, but not as a holistic training program in the general population. To contribute to closing this research gap, we developed the Positive Emotions Training (PoET) which refers to the promotion of several positive psychology constructs and the improvement of mental health. The development of PoET was based on existing literature on positive psychology treatments as well as long-term discussions among the research group about how to promote respective constructs. For the training, an online format was chosen so that it could be conducted in larger groups despite the prevailing COVID-19 pandemic.We decided to test PoET as an intervention and the methodical approach in a feasibility study. The main goal of this study was to evaluate the practicability of PoET and examine whether it might be an effective intervention to promote mental health. First, we hypothesized that participants receiving PoET would show a significant increase in positive mental health factors one month after the second training day (Hypothesis 1). Second, we expected to find a significant decrease in depression, anxiety, and stress symptoms one month after the second training day (Hypothesis 2). Third, we hypothesized participants in the control group not to show any significant changes in this regard (Hypothesis 3). In addition, we examined whether there might be a significant increase in optimism, gratitude, happiness, resilience, and self-efficacy for participants of PoET in comparison to the control group. | PMC10426081 |
Methods | PMC10426081 |
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Recruitment | mental illness | To advertise the PoET, a flyer was distributed on social media and two articles were published in newspapers. Those interested could contact an email address to receive further information about the training and to apply. Participants for the control group were also recruited via social media. As PoET is a low-threshold intervention, there were no exclusion criteria for participation and no screening for mental illness before the start. Any person older than 18 years could participate. Psychology students could get VPN hours as compensation. Overall, nearly 600 people were interested in participating. Because the number of psychologists to conduct training sessions was limited, not everyone got the possibility to participate. Places in the intervention study were not randomly allocated, but according to the principle “first-come, first-served”. Participants of the control group did not receive any treatment initially, but were offered to participate in one of the next PoETs. | PMC10426081 |
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Participants | A total of | PMC10426081 |
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PoET | Since we were interested in the specific outcome effects of the training, only participants who took part at both training days and completed all data sets were considered in the analyses. | PMC10426081 |
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Control | The final sample consisted of The two groups differed significantly in their age ( | PMC10426081 |
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Development and description of PoET | humor | POSITIVE | For the development of PoET, a systematic literature analysis was first carried out within the context of the project “On the Role of Positive Emotions in the Promotion of Mental Health - An Experimental Study” to collect various positive psychology constructs and exercises for the promotion of positive emotion experience. Over a period of three years, these results were discussed in context of the Annual Congress for Psychotherapy in Bochum and a list of about 20 important positive psychological constructs was developed. A further literature analysis of the existing possibilities for promoting these constructs was conducted. Based on their potential regarding feasibility and effects on mental health, eleven constructs were selected by the team and included in the training. Exercises from already existing literature were compiled and own exercises were developed afterwards. After that, the exercises and formal aspects of the training were tested in a pilot phase in January 2022. In the context of this pilot phase, an exemplary training session was conducted with a group of volunteers and feedback was collected as well as integrated into the general PoET concept.The PoET conducted in spring 2022 consisted of two training days of 3.5 h each, one week apart. The training was carried out online via Zoom in groups of about 30 to 35 people and was led by two psychologists who were also part of the research team. A total of eleven lessons on positive psychology constructs were conducted: happiness, hope, optimism, humor, self-efficacy, gratitude, flow, meaningfulness, forgiveness, spirituality, and resilience. At the beginning of each lesson, the participants received a short theoretical input on the respective positive psychology construct. After that, exercises were carried out either in the large group, in groups of three to six people in breakout rooms or alone. Afterwards, there was a short discussion on how the participants experienced each respective exercise. Finally, participants were given at-home-exercises. In addition, the participants received a booklet with descriptions of the exercises and space for notes. During the training days, a PowerPoint presentation was used as visual support. Between the two training days, there was a so-called 7-day challenge which should motivate the participants to complete the exercises at home. Table
PoET lessons and respective exercises
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Measures | At all three assessment points, participants in the PoET and control group completed a battery of questionnaires. Socio-demographic data, questions about the experience of flow, current mood and stress, and exercise practice at home (in the PoET group) were assessed. In addition, the battery contained the German versions of ten validated questionnaires. The following seven questionnaires were relevant for this study. | PMC10426081 |
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DASS-21 | Depression, Anxiety | The Depression Anxiety Stress Scale [ | PMC10426081 |
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PMH | POSITIVE | The Positive Mental Health Scale [ | PMC10426081 |
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LOT-r | Orientation Test-revised [ | The Life Orientation Test-revised [ | PMC10426081 |
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GQ-5 | The Gratitude Questionnaire [ | PMC10426081 |
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SWLS | The Satisfaction with Life Scale [ | PMC10426081 |
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NGSE | The New General Self-Efficacy Scale [ | PMC10426081 |
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