bluuebunny/crossref_metadata_2025 huggingface dataset:
Gastrointestinal morphology, tight junction gene expression, and microbiota of the Ossabaw obese pig model (816.1)
Authors: Ahamed Kathryn , Holscher Hannah , Kil Dong Yong , Newell‐Fugate Anne , Krisher Rebecca , Swanson Kelly
Published: June 2021

Obesity is characterized by chronic low‐grade inflammation, with reduced gastrointestinal (GI) barrier function hypothesized to be a contributing factor. Animal models are invaluable for understanding disease pathophysiology, with porcine models offering several advantages over rodents, including anatomical similarities to humans. A pilot study was undertaken to characterize GI differences in obese Ossabaw pigs vs. lean controls. Ossabaw gilts (n=8) were fed a high‐fat, high‐sugar diet ad libitum until 12 months of age and compared to lean Yorkshire controls (n=9). Plasma inflammatory markers (TNF‐α, IL‐6, and LPS‐binding protein) and intestinal morphology, tight junction protein gene expression, and mucosal microbiota populations were assessed. Preliminary results indicate that LPS‐binding protein concentrations tended to be greater (P=0.08) in obese vs. lean pigs (6233 vs.3802 µg/L). Blood TNF‐α concentrations were numerically, but not significantly increased in obese vs. lean pigs (39 vs. 29 ng/L; P=0.24). Obese pigs had greater (P=0.03) cecal crypt depth (494 vs. 430 µm) and tended to have greater (P=0.09) ileal villus height (477 vs. 400 µm) compared to lean controls. Our findings suggest a correlation between GI architecture, systemic inflammation, and obesity in high‐fat, high‐sugar fed Ossabaw pigs, supporting its use as a model of human obesity and metabolic syndrome.

Grant Funding Source: Supported by USDA Hatch #ILLU‐538‐384
Effect of Actigen and threonine interaction on the development of gut immunity of nursery pigs (816.3)
Authors: Graugnard Daniel , Samuel Ryan , Smith Allison , Good Lindsay , Jolliff James , Mahan Donald , Brennan Kristen
Published: June 2021

Weaned piglets were allocated to six treatment groups (n = 6/treatment) in a 2 x 3 factorial design experiment: Actigen (0 or 400 g/T), true ileal digestible (TID) threonine (Thr):lysine (Lys) ratio (57 [low], 67[recommended], and 77 [high]). After 6 wk of feeding, duodenum, jejunum and ileum tissues were sampled for qPCR analysis. Expression of cytokines was reported relative to the control (no Actigen + 67 TID Thr:Lys). In jejunum, interleukin 6 (IL6) was down‐regulated (P < 0.05) with Actigen + 57 or 77 TID Thr:Lys, (‐5.89 and ‐4.14‐fold, respectively). In ileum, interleukin 1 beta (IL1B) was down‐regulated (P < 0.05) with Actigen + 57 and 77 TID Thr:Lys (‐1.35 and ‐1.75‐fold, respectively). In duodenum, IL1B was up‐regulated (P < 0.05) (1.89‐fold) with Actigen +57 TID Thr:Lys. In duodenum and jejunum, tumor necrosis factor (TNF) was up‐regulated (P < 0.05) with Actigen + 67 TID Thr:Lys (1.21‐fold and 1.33‐fold, respectively). In duodenum, IL6 (‐1.51‐fold) and TNF (1.32‐fold) differed (P < 0.05) in response to no Actigen +57 and 77 TID Thr:Lys, respectively. Relative to the control, Actigen and Thr, in combination, were associated with decreased mRNA levels of the inflammatory cytokines IL6, IL1B in the intestine of weaned pigs.
Estrogen plays an important role in intestinal calcium transport (816.5)
Authors: Tokuno Yuko , Zhao Yitong , Johnson Sarah , Pourafshar Shirin , Arjmandi Bahram
Published: June 2021

Objective: To determine the presence of estrogen receptor β (ERβ) in the small and large intestines between ovariectomized and sham‐operated rats via immunocytochemistry; and to investigate the role of estrogen in calcium uptake in the duodenum. Methods: Twelve 12‐week old female Sprague‐Dawley rats were either ovariectomized (Ovx; n=6) or sham‐operated (Sham; n=6) and sacrificed three weeks later. Segments of the small intestine (duodenum, jejunum, and ileum) and large intestine (cecum) were removed for immunocytochemistry analysis. Duodenal cells were harvested for measurement of calcium transport. Results: Both Ovx and Sham groups were found to have ERβ immunoreactivity in all segments of the intestine although the extent was greater in the Sham group when compared to the Ovx group. The presence of ERβ was more pronounced in the duodenum than the jejunum, ileum, or cecum. The rates of calcium uptake in the Sham group were significantly (P = 0.028) greater compared to the Ovx group. Conclusion: The findings of the present study suggest that estrogen is essential for the regulation of duodenal calcium absorption and that menopause is associated with a decline in calcium absorption which may play a role in postmenopausal bone loss.

Grant Funding Source: Supported by Jumonji University and Collage of Human Sciences, Florida State University
Genome‐wide DNA methylation in whole blood and CD16+ cells in response to chronic folic acid supplementation in women of child‐bearing age (817.1)
Authors: Shade Deanna , Park Hea Jin , Hausman Dorothy , Hohos Natalie , Meagher Richard , Kauwell Gail , Smith Alicia , Bailey Lynn
Published: June 2021

Folate is a key source of one‐carbon groups for DNA methylation; but studies on DNA methylation response to folic acid (FA) supplementation show inconsistent results. We hypothesize that measuring DNA methylation in whole blood (WB), which contains a mixture of distinct leukocyte cell types, may have confounded the results of previous studies. Therefore, the aim of this pilot study was to determine if a single cell type could serve as a more reliable epigenetic reporter. The study was performed in normal weight (BMI 18.5 – 24.9 kg/m2) women (18 – 35 y; n = 12), with blood samples taken before and after eight weeks of supplementation with 800 ug/day of FA. CD16+ cells (CD16) were isolated using antibody‐bound magnetic beads. DNA methylation patterns from WB and CD16 cells were measured across >485,000 CpG sites using the Infinium Human Methylation450 Bead Chip. For each CpG site, the proportion of DNA methylation that changed over the 8 week intervention was evaluated by fitting a separate linear model. Genome‐wide analysis indicated changes in 7887 vs. 8482 CpG sites in response to supplementation in WB and CD16, respectively; with DNA methylation decreased in 77.2% (WB) or 66.3% (CD16) of these sites. Results suggest that there are differences in methylation response in WB and CD16 after FA supplementation. The implication of these methylation differences are being explored in our on‐going gene enrichment analyses.

Grant Funding Source: Funded in part by HATCH #GEO00706, #GEO00707 and the Obesity Initiative at the University of Georgia
TETs are involved in adipogenic differentiation via Hif‐1a mediated regulation of Pparg (817.5)
Authors: Yoo Yeongran , Park Yoon Jung
Published: June 2021

TET proteins are a‐Ketoglutarate dependent hydroxylases, which convert 5‐methylcytosine to 5‐hydroxymethylcytosine in DNA. They have been described as key modulators for DNA methylation pattern in locus‐specific and genome‐wide levels. In this study, we showed that the role of TET during 3T3‐L1 cell differentiation in vitro. We found that the Tet1 and Tet2 were induced in 3T3‐L1 cells during adipocyte differentiation, and this pattern paralleled that of other adipogenic gene markers like Pparg and Glut4. RNA‐interference suppression of Tet1 in 3T3‐L1 cells interfered the adipocyte differentiation by downregulating Pparg gene expression, a master regulator for adipoctye specific genes. Furthermore, the depletion of Tet1 led to the elevation of Hif‐1a gene expression, which is a transcription factor for hypoxia regulated genes that has been shown to block adipogenesis by transcriptional inhibition of Pparg. Intriguingly, Hif‐1a has a binding site for both Tet1 and Tet2 in its promoter, suggesting that TETs are possible transcriptional regulators of Hif‐1a. The results demonstrated that TETs, a‐Ketoglutarate‐dependent epigenetic modulator, are involved in adipogenic differentiation via Hif1‐a‐mediated regulation of Pparg.

Grant Funding Source: Supported by Korean National Research Foundation of Korea(22A20130012143)
Evaluation of candidate reference genes for quantitative real‐time PCR analysis in a rat model of dietary iron deficiency (818.15)
Authors: Fiddler Joanna , Hill Jacob , Lucas Edralin , Smith Brenda , Clarke Stephen
Published: June 2021

Quantitative real‐time polymerase chain reaction (qPCR) is a commonly used, reliable, and efficient method for quantitation of gene expression. Due to the increased use of qPCR in examining nutrient‐gene interactions, it is important to develop standardized approaches to analyzing and presenting qPCR data. (Bustin SA, et al, Clinical Chemistry 2009, 55(4): 611‐622). One of the most common approaches to normalizing expression data involves the use of reference genes. Thus, the choice of reference genes (RGs) has a major influence on calculating qPCR results. Although RGs are commonly used, relatively little attention is given to the appropriateness of RG selection for an experimental condition or specific tissue/cell. Using qPCR, we examined the stability of gene expression between iron‐deficient and control rats in both liver and skeletal muscle to determine the suitability of twelve potential RGs. Using Normfinder and Bestkeeper software, we then compared the relative stability of RGs between ID and C animals for either liver or skeletal muscle. Next we compared the stability of RGs to examine suitability of RGs across tissues. Both methods were combined and used to rank the RGs according to overall suitability. We provide evidence that commonly selected RGs (β‐actin and Gapdh) may be more susceptible to variation than less commonly used RGs that exhibit increased stability regardless of treatment (e.g., Rpl19 and Rpl27). These results indicate that the selection and use of RGs should be empirically determined and may not be similar across experimental conditions or biological tissues.
An interaction effect between glucokinase gene variation and carbohydrate intakes modulate the plasma triglyceride response to a fish oil supplementation (818.3)
Authors: Bouchard‐Mercier Annie , Rudkowska Iwona , Lemieux Simone , Couture Patrick , Vohl Marie‐Claude
Published: June 2021

A large inter‐individual variability in the plasma triglyceride (TG) response to fish oil consumption has been observed. A single‐nucleotide polymorphism (SNP) within glucokinase (GCK) gene has been associated with TG concentrations. Objective: To investigate the gene‐diet interaction effects between SNPs within GCK gene and dietary carbohydrate intakes on the plasma TG response to a fish oil supplementation. Methods: Two hundred and eight participants were recruited in the greater Quebec City area and completed a 6‐week fish oil supplementation (5g fish oil/day: 1.9‐2.2g EPA and 1.1g DHA). Thirteen SNPs within GCK gene were genotyped using TAQMAN methodology. Results: A gene‐diet interaction effect on the plasma TG response was observed between rs741038 and carbohydrate intake when age, sex and BMI were included in the model (p=0.008). In order to compare the plasma TG response between genotypes, participants were divided according to median carbohydrate intakes. Homozygotes of the minor C allele of rs741038 with carbohydrate intakes >48.6% had a greater decrease in their plasma TG concentrations than homozygotes C/C with carbohydrate intakes 蠄48.6% (p=0.002) and also than the other genotypes either with high or low CARB. Conclusion: The plasma TG response to a fish oil supplementation may be modulated by gene‐diet interaction effects involving GCK gene and carbohydrate intakes.

Grant Funding Source: Supported by CIHR ‐ (MOP229488)
Effects of chemoprotective diets on crypt adult stem cells û the cells of origin of colon cancer (819.1)
Authors: Kim Eunjoo , Davidson Laurie A. , Patil Bhimanagouda S. , Jayaprakasha Guddadarangavvanahally K. , Callaway Evelyn S. , Turner Nancy D. , Chapkin Robert S.
Published: June 2021

The majority of colon tumors are triggered by aberrant Wnt signaling in intestinal stem cells, which is an extremely efficient route towards initiating intestinal cancer. Curcumin, a component of turmeric, and n‐3 polyunsaturated fatty acids (PUFAs), mainly found in fish oil, exert chemoprotective and anti‐inflammatory properties in part by modulating apoptotic, Wnt, and/or NFκB related pathways. However, the effects of diet on colonic stem cells are unknown. Therefore, we determined whether the combination of fish oil and curcumin reduced colon cancer risk by promoting targeted apoptosis of damaged colonic stem cells at the initiation stage of cancer. Lgr5‐EGFP‐IRES‐creERT2 knock in mice (n=10 per treatment) were fed diets containing corn oil ± curcumin (CO±C) or fish oil ± curcumin (FO±C) for 3 weeks. Apoptosis, cell proliferation and DNA damage in Lgr5+ stem cells were quantified 12 h post carcinogen (azoxymethane) injection. Interestingly, mice fed FO diets exhibited 15% fewer stem cells per crypt and an enhanced resistance to DNA damage, with 52% fewer damaged stem cells. For those stem cells exhibiting DNA damage, FO feeding enhanced targeted apoptosis by 24% compared to CO diets. In contrast, curcumin had no significant effects on stem cell responses. These novel results demonstrate the chemoprotective ability of external dietary cues to impact intestinal stem cell responses following carcinogen exposure.
FOXO1 activates glutamine synthetase gene in mouse skeletal muscles (820.10)
Authors: Kamei Yasutomi , Hattori Maki , Hatazawa Yukino , Kasahara Tomomi , Kitamura Tadahiro , Ogawa Yoshihiro
Published: June 2021

Skeletal muscle is the largest organ in the human body, and is a reservoir of energy in the form of protein (amino acids). FOXO1 is a forkhead‐type transcription factor, whose expression increases during conditions such as protein degradation, starvation and exercise. We have previously generated transgenic mice overexpressing FOXO1 specifically in skeletal muscles (FOXO1‐Tg mice), which exhibited muscle atrophy. In this study, we found that glutamine levels and glutamine synthetase (GS) expression increased in skeletal muscles of FOXO1‐Tg mice. During fasting, FOXO1 and GS expression markedly increased in wild‐type mice, but the increase in GS expression was attenuated in mice with a skeletal muscle‐specific knockout of FOXO1 (FOXO1‐KO mice). In a transient transfection reporter assay, FOXO1 dose dependently activated the GS reporter construct. Moreover, FOXO1 was strongly recruited to the 3’ region of GS in C2C12 myoblasts, as analyzed by ChIP assay. These results suggest that FOXO1 directly upregulates GS expression. Meanwhile, blood ammonia levels were significantly increased in FOXO1‐KO mice and inversely correlated with suppressed GS expression, which is considered to be involved in ammonia clearance in skeletal muscles. In conclusion, FOXO1 in skeletal muscles may play a role in amino acid metabolism, particularly in glutamine metabolism.
Excess leucine intake effects on asparagine synthetase and 3‐phosphoglycerate dehydrogenase expression in the liver of rats fed a low‐protein diet (820.13)
Authors: Yoshimura Ryoji , Mizushige Takafumi , Ohinata Kousaku , Kamei Yasutomi , Kanamoto Ryuhei
Published: June 2021

De novo synthesis of dispensable amino acids such as asparagine and serine is crucial for cell growth and function. We found that asparagine synthetase (AS), 3‐phosphoglycerate dehydrogenase (PHGDH, key enzyme for serine biosynthesis), and serine dehydratase (SDH, serine degradative enzyme) expression in rat liver were tightly regulated in response to the protein requirement. Thus, AS and PHGDH were induced through a low‐protein diet, below rat’s protein requirement, whereas SDH was induced through a high‐protein diet, above their requirement. However, we observed that SDH was also induced by leucine ingestion. Excess leucine intake through a low‐protein diet causes severe growth retardation in rat. We here examined the effect of dietary leucine on AS, PHGDH, and SDH expression with regard to growth retardation. Male rats were divided into 12 groups and fed for one week a diet containing low, moderate or high protein. Different levels of leucine (0, 2, 4, and 8%) were added to the diets. The results demonstrated that AS and PHGDH expression was dose‐dependently suppressed in rat liver, whereas SDH expression increased when rats were maintained on a low‐protein diet. Body weight and food intake concomitantly decreased. Valine and isoleucine affected neither gene expression nor food intake and body weight. These results suggested that leucine suppressed AS and PHGDH expression and could cause growth retardation.
The effect of exercise intensity on sweat amino acid excretion (820.14)
Authors: Stone Michael , Buono Michael , Kern Mark
Published: June 2021

We examined the relationships between sweat gland amino acid (AA) secretion and exercise intensity as well as sweat and plasma AA concentrations as functions of AA molecular weight (MW). Ten subjects underwent 4 trials including a maximal graded exercise test and 3 submaximal bouts (low: 45% VO2max, moderate: 60% VO2max and high: 75% VO2max) for 30 min. Sweat was collected throughout exercise using macroducts placed on the forearm and blood was collected before and after exercise. Sweat AA excretion (pmol/cm2/min) was significantly greater (p<0.05) between the higher workloads and the 45% workload, but were not greater at the 75% versus 60% workload despite higher sweat rates, suggesting dermal conservation during high intensity exercise. With the exception of alanine, which was increased during the 75% workload, plasma concentrations of AA decreased during the 75% bout. Similar effects were not detected during the lower intensity bouts. Polynomial regression revealed a negative curvilinear relationship between plasma AA concentration and molecular weight (r = ‐0.71, p = 0.002) as well as sweat AA excretion rate and MW (r = ‐0.75, p = 0.004) implicating an influence of MW on AA handling.
Creatine supplementation to total parenteral nutrition increases muscle and organ creatine and lowers liver cholesterol in Yucatan miniature piglets (820.17)
Authors: Dinesh O , Bertolo Robert , Brunton Janet
Published: June 2021

In TPN‐fed pigs, we previously demonstrated that tissue creatine concentrations were proportionate to the IV arginine intake, suggesting that arginine availability limited creatine synthesis. TPN is devoid of creatine, so the requirement must be met entirely by de novo synthesis requiring arginine, glycine and methionine as a methyl donor. L‐arginine:glycine amidinotransferase (AGAT) converts arginine and glycine into guanidinoacetic acid (GAA); GAA is methylated via Guanidinoacetate methyltransferase (GAMT) to form creatine. The addition of creatine to TPN may spare arginine and methionine for protein synthesis and other metabolic processes. Piglets (3‐5 d old, N = 14) were fed complete TPN with (CRE, 0.1 g/kg/d) or without creatine (CON) for 14 d. The CRE group had lower kidney and pancreas AGAT activity and lower plasma GAA concentration (P < 0.01) demonstrating a down‐regulation of creatine biosynthesis. CRE pigs also had greater creatine in plasma, liver, kidney and pancreas (P < 0.01). Total creatine in muscle was also higher in CRE (P = 0.05); however, brain creatine was not affected by treatment. There was no difference in liver weight or triglyceride concentration, but total liver cholesterol was ~50% lower in the CRE piglets compared to CON (P < 0.01). Altered lipid metabolism suggests that creatine may have a role in ameliorating TPN‐induced liver damage. The neonatal brain appears to have priority for creatine when supply is limited, but the addition of creatine to TPN may be necessary to support total body creatine accretion in rapidly growing neonates.

Grant Funding Source: Supported by Janeway
Evaluating the effect of 20‐hydroxyecdysone on leucine signaling in the skeletal muscle and liver of male rats (820.7)
Authors: Anthony Tracy , Mirek Emily , DeOliveira Christopher , Bargoud Albert , Marella Dean , Phillipson‐Weiner Lindsey , Graf Brittany , She Pengxiang , Kuhn Peter , Wilson Gabriel , Raskin Ilya
Published: June 2021

Phytoecdysteroids such as 20HE are ingredients in various nutritional supplements marketed to enhance physical performance and lean body mass. Recent in vitro studies suggest 20HE may activate protein synthesis via a phosphoinositide‐3‐kinase‐Akt signaling mechanism. To confirm these findings in vivo and evaluate downstream activation of mammalian target of rapamycin complex 1 (mTORC1) signaling, overnight fasted 4‐6 week old male rats (n=5‐6 per group) were randomized to one of three study designs: 1) gavaged with 0, 10, 50, 200 mg/kg 20HE and euthanized 30 min post‐gavage; 2) gavaged with 200 mg/kg 20HE or excipient and euthanized at 30 min, 60 min, 120 min, 240 min post‐gavage, and 3) administered excipient or 200 mg/kg 20HE alone or in combination with 1.35 g/kg L‐leucine and euthanized 30 min post‐gavage. Independent evaluation of 20HE confirmed purity as reported by Sigma before solubilization in 3% DMSO or 70% Labrasol® as excipient. Phosphorylation of Akt at Thr308 and Ser473, mTOR at Ser2448, ribosomal S6 kinase (S6K1) at Thr389, and eukaryotic initiation factor 4E binding protein 1 (4E‐BP1) at Thr 37/46 were evaluated by immunoblot. Leucine administered in 3% DMSO or Labrasol robustly increased phosphorylation of mTOR, S6K1 and 4E‐BP1 but not Akt in muscle and liver. In contrast, at all doses and times, 20HE did not significantly increase phosphorylation of Akt, mTOR, S6K1 or 4E‐BP1 as compared to excipient controls. Furthermore, 20HE dampened mTORC1 signaling by leucine in both muscle and liver. These data do not support the idea that anabolic effects of 20HE are acutely mediated by mTORC1.

Grant Funding Source: Supported by NJAES and USDA Multistate NC1184
Effects of a 7‐day military training exercise on whole‐body protein turnover: an observation of military‐specific protein requirements (820.9)
Authors: Pasiakos Stefan , Murphy Nancy , Margolis Lee , Gundersen Yngvar , Martini Svein , Montain Scott , Young Andrew
Published: June 2021

Physically intense military training may diminish whole‐body protein retention and increase dietary protein requirements. This study characterized the effects of military training on whole‐body protein turnover in 21 Soldiers participating in a 7‐day winter training exercise that culminated in a 3‐day, 54 km ski march. Energy expenditure was assessed during training using doubly labeled water (D218O), and protein turnover, synthesis, breakdown, and net balance were measured at baseline (BL), day 4 (PRE, prior to the ski march), and day 7 (POST, following the ski march) using 15N‐glycine. Energy (kcal·d‐1) intake increased (P < 0.05) PRE ([mean ± SD] 3115 ± 437) to POST (3415 ± 977), while protein (g·kg‐1·d‐1) intake remained constant (PRE, 1.6 ± 0.4; and POST, 1.7 ± 0.5). Although energy expenditure (kcal·d‐1) increased (P < 0.05) PRE (5171 ± 485) to POST (7190 ± 600), turnover remained steady during training (group mean ± SD, 1.4 ± 0.3 g·N‐1·kg‐1·d‐1). Synthesis and breakdown were not influenced (P > 0.05) by training. Net balance (g·kg‐1·d‐1) was maintained, as levels were similar (P > 0.05) between BL (0.04 ± 0.3), PRE (0.01 ± 0.5), and POST (‐0.4 ± 1.0). Despite high energy expenditure and resultant energy deficit, whole‐body protein was conserved. These data suggest that protein requirements during short‐term, metabolically demanding military training are likely twice the recommended dietary allowance.

Grant Funding Source: Supported by USAMRMC and FFI
Brain docosahexaenoic acid accretion is greater when supplied as phosphatidylcholine than as triacylglycerol in piglets (821.11)
Authors: Liu Lei , Bartke Nana , Van Daele Hans , Lawrence Peter , Qin Xia , Park Hui , Kothapalli Kumar , Windust Anthony , Wang Zhe , Brenna J.
Published: June 2021

Introduction. Docosahexaenoic acid (DHA) is the most abundant n‐3 PUFA in the brain. Preformed DHA is more efficacious for brain DHA accretion than precursors, but the relative efficacy of DHA bound to phospholipid (PC) and/or triacylglycerol (TAG) has not been measured yet. Our aim was to determine the relative efficacy of DHA when provided in formula to the growing piglet as a dose of 13C‐DHA bound to the sn‐2 positions of either PC or TAG.

Methods. Piglets (n=8 per group) were assigned to two identical formula diets from day 3 of life and provided with a single oral dose of TAG‐13C‐DHA or PC‐13C‐DHA at 16 days of life. At day 23, selected piglet organs were analyzed for 13C‐DHA and other fatty acid metabolites.

Results. The PC‐13C‐DHA was 1.9‐fold greater in brain gray matter than TAG‐13C‐DHA, and was similarly more efficacious in synaptosomes, retina, liver, and RBC. Liver labeling was by far the greatest implying initial processing in that organ followed by export to other organs, and in turn that transfer from gut to bloodstream to liver in part drove the relative efficacy for tissue accretion. Apparent retroconversion to 22:5n‐3 was more than 2‐fold greater for PC and was much more prominent in neural tissue than in liver or RBC.

Conclusions. These data directly support greater efficacy for PC as a carrier for LCPUFA compared to TAG, consistent with previous studies of arachidonic acid and DHA measured in other species.

Grant Funding Source: Supported by NIH R01 AT007003
Association between glucose fructose oxido‐reductase domain containing 2 (GFOD2, rs12449157) polymorphism and blood lipids in hypercholesterolemic Mexican subjects (821.4)
Authors: Flores Adriana , Guevara Martha , Castaño‐Jameson Marisol , Vazquez Natalia , Ayala Brenda , Medina Isabel , Orozco Ximena , Jimenez Edith , Jimenez Edith , Aguilar Miriam , Estrada Isabela , Tovar Armando , Torres Nimbe
Published: June 2021

It has been demonstrated that almost 28% of the Mexican population presents the GFOD2 SNP rs12449157 that is associated with lipids. Thus, the aim of this study was to assess the presence of this SNP and biochemical parameters in subjects with hypercholesterolemia. We studied 696 Mexican subjects (139 men, 557 women). The number of participants with hypercholesterolemia was 271, from these, 94 subjects (33%) had the GFOD2 SNP. Interestingly, subjects with hypercholesterolemia and the G allele showed lower serum HDL‐C concentration than those with the common allele (p<0.008), higher total cholesterol and higher LDL‐C than the common allele group (p<0.0001). Also, increase in dietary intake of lipids in subjects with the G allele shows higher levels of LDL cholesterol. Thus, the presence of this polymorphism increase the risk to present cardiovascular disease.

Grant Funding Source: Supported by Medix (to NT) and , CONACYT 181685 (to MG)
Effects of dairy ingredients on gut microbiome biogeography in mice (822.11)
Authors: Ananda Baskaran Sangeetha , Price Tara , Wu Xiuzhi , Walzem Rosemary
Published: June 2021

We tested the effects of purified diets (PD) containing isolated soy protein (ISP), dried whole milk powder (DWMP), milk fat globule membrane (MFGM) or milk protein concentrate (MPC) on gut microbiome biogeography of C57BL/6J mice. Isocaloric, isonitrogenous PDs provided 20 energy % (EN%) protein, 50 EN% carbohydrate and 30 EN% fat. Mice were fed a chow diet for 1 wk then assigned to a test diet for 13 wk, n = 10 per group. Mice had similar growth, feed intake and intestinal transit times. Feces were collected before and at the end of test diet feeding. Samples of lumenal (L) and adherent (A) microbiota were collected from the distal small intestine (DSI) and descending colon (DCS). Ion Torrent sequencing technology determined microbiota composition (MrDNA, Sweetwater, TX). Paired t‐tests for Chao1 & Shannon indexes showed diversity decreased from DSC‐L > feces > DSI‐A & DSC‐A > DSI‐L (p<0.02); fecal diversity was not altered by switching mice from non‐purified chow to PDs. Firmicutes dominated in all gut regions being 蠅 60% of bacterial reads (BR). Bacteriodetes was ~20% BR in feces & DSC‐L, ~6.5% BR in DSI‐L, DSC‐A or DSI‐A (p < 0.01). Proteobacteria was ~21% BR in DSC‐A, and ~1.4% BR in all other regions (p < 0.001). Verrucomicrobia rose from ~0.64% BR to ~10.4% BR (p < 2.7 x 10‐8) with PD, as did Actinobacteria (0.4% BR vs. 7.0% BR, p <0.03), while Tenericutes fell from ~ 6.3% to 1.0% of BR (p < 1x 10‐7). Diet alters microbiome biogeography.

Grant Funding Source: National Dairy Council project #1120 administered by Dairy Research Institute
Is the omega‐3 index a valid marker of intestinal membrane phospholipid EPA+DHA content? (822.2)
Authors: Gurzell Eric , Wiesinger Jason , Harris William , Fenton Jenifer
Published: June 2021

Despite numerous studies investigating n‐3 fatty acids (FA) and inflammatory bowel diseases, the extent to which dietary n‐3 FAs incorporate in gastrointestinal (GI) tissues is unknown. In this study, mice were fed three diets with increasing percent of energy (%en) derived from EPA+DHA, reflecting recommended intakes of fish/fish oil. We analyzed the FA composition of phospholipids extracted from RBCs, plasma, and GI tissues. We observed that the 0.001%en EPA+DHA diet was sufficient to significantly increase the omega‐3 index (RBC EPA+DHA) after 5 week feeding. The baseline EPA levels were 0.2‐0.6% across all tissues increasing to 1.6‐4.3% in the highest EPA+DHA diet; these changes resulted in absolute increases of 1.4‐3.9% EPA across tissues. The baseline DHA levels were 2.2‐5.9% across all tissues increasing to 5.8‐10.5% in the highest EPA+DHA diet; these changes resulted in absolute increases of 3.2‐5.7% DHA across tissues. We identified strong (r > 0.91) and significant (p < 0.001) linear correlations between the omega‐3 index and plasma/GI tissue EPA+DHA content, suggesting that the omega‐3 index reflects the relative amounts of EPA+DHA in GI tissues. These data demonstrate that the GI tissues are highly responsive to dietary LCPUFA supplementation and that the omega‐3 index can serve as a valid biomarker for assessing dietary EPA+DHA incorporation in GI tissues.

Grant Funding Source: Supported by NIH R03CA162427 (to JIF)
The source of dietary fat modulates the intestinal microbiota and liver metabolite pattern (822.6)
Authors: Solano‐Aguilar Gloria , Conchas Andrew , Molokin Aleksey , Jang Saebyeol , Lakshman Sukla , Santin‐Duran Monica , Sikaroodi Masoumeh , Gillevet Patrick , Urban Joseph
Published: June 2021

The source of dietary fat modulates the intestinal microbiota and liver metabolite pattern Gloria Solano‐Aguilar1, Andrew Conchas1, Aleksey Molokin1, Saebyeol Jang1, Sukla Lakshman1, Monica Santin‐Duran1, Masoumeh Sikaroodi2, Patrick Gillevet2, Joseph Urban1.

1USDA, ARS, Beltsville Human Nutrition Research Center, Diet, Genomics, and Immunology Laboratory, Beltsville, MD 20705; 2Microbiome Analysis Center, George Mason University, Manassas, Virginia 20110 The goal of this study was to characterize the intestinal microbiome and host metabolism consequent to feeding an obesogenic diet with different fat sources. We examined the effect of anhydrous milk fat (AMF) and palm oil (PO) on the liver metabolite profile and composition of the intestinal microbiota in juvenile pigs fed either a 10.8% low fat (LF) or 39.5% high fat (HF) diet. Thirty two pigs were randomly allocated to dietary treatments: 1) LF‐PO, 2) LF‐AMF, 3) HF‐PO and 4) HF‐AMF for 18 weeks. Fecal samples were collected every three weeks to determine changes in total coliforms, and commensal Bifidobacterium, Lactobacillus and Bacteroides species by RT‐PCR. Proximal colon contents and liver samples were also collected at necropsy and used, respectively, for metagenomic analysis (Multitag Pyrosequencing) and global metabolite profile analysis (GC/MS and LC/MS/MS platforms) and compared across dietary treatment groups. The only affect on the microbiome was an increase in total coliforms in pig fed HF‐PO. Different metabolite clusters were identified between pigs fed the HF and LF diets , and pigs fed HF‐PO and HF‐AMF diets suggesting significant differences in metabolites induced by the amount and type of fat in the diet. Our data suggested a fat source dependent effect on microbiota and liver metabolite composition. The use of AMF may constitute a better source of dietary fat to prevent the increase in pro‐inflammatory responses positively associated with increases in intestinal coliforms.

Grant Funding Source: Supported by DRI #1101
Intestinal inflammation in response to omega 3 and 6 intake (822.8)
Authors: Campbell Sara , Wisniewski Paul , Joseph Laurie , Lightfoot Stan
Published: June 2021

The consumption of omega‐3 and omega‐6 essential fatty acids in Western diets is thought to have changed markedly during the 20th century which may contribute to rising obesity rates. We aimed to understand the impact n‐3 and n‐6 rich diets have on intestinal morphology. For this study, 36 male C57 mice were randomly divided into 4 groups (n of 9) and after acclimation were fed diets for 3‐months as follows: Group 1 (10% fat 1:1 omega 6:omega 3 ratio); Group 2 (10% fat 16:1 omega 6:omega 3 ratio); Group 3 (45% fat 1:1 omega 6:omega 3 ratio); and Group 4 (45% fat 16:1 omega 6:omega 3 ratio). Food intake was monitored every other day and body weight every week. Upon sacrifice duodenal sections were harvested for IHC analysis including H&E, COX‐2 and trichrome with Gomorra blue. Histological analysis revealed that diets high in both omega‐3s and 6s manifested short, fat and inflamed villi that had reduced cytoplasm in the columnar cells. COX‐2 expression supported these findings and showed that as diets increased in percent fat staining in small cells which are PMN's, lymphs or small macrophages, increased from positive cells every 3‐4 high power fields (10% 1:1) to a positive cell every high power field (45% 16:1). Finally, excessive mucus and collagen presence was noted in both 45% fat diets in the crypts of Leiberkuhns. Increased intake of omega 3s and/or 6s in the diet, regardless of percentage total fat, significantly disrupts intestinal morphology and increases inflammation.
The effects of pork vs. chicken/fish in a DASH diet on blood pressure control (823.4)
Authors: Sayer Richard , Chen Ningning , Wright Amy , Campbell Wayne
Published: June 2021

Hypertension is a major, modifiable risk factor for cardiovascular and kidney disease and mortality that is improved by the Dietary Approaches to Stop Hypertension (DASH) diet. The DASH diet emphasizes increased consumption of fruits/vegetables, whole‐grains, lowfat dairy, and poultry/fish and reduced intakes of sodium and red meats (including pork). In a randomized crossover study, 13 women and 6 men, aged 61±7 y (mean±SEM), BMI 31.2±1.7 kg/m2, with high blood pressure (BP) consumed the DASH diet for 2, 6‐wk periods (with a 4‐wk diet washout) with either chicken/fish (DASH‐CF, control diet) or lean pork (DASH‐P) as the major protein source (55% of protein intake). Systolic (S) and diastolic (D) BP were assessed manually (fasting, morning, seated) and with a 24h BP monitoring system on 3 days prior to and at the end of each diet period. After DASH‐CF, manual SBP and DBP decreased 6±2 and 6±1 mmHg, respectively, and 24h SBP and DBP decreased 9±2 and 5±1mm Hg. After DASH‐P, manual SBP and DBP decreased 8±2 and 5±1 mmHg, and 24h SBP and DBP decreased 8±2 and 4±1 mmHg. All changes in BP were statistically confirmed at p<0.05 and were not different for DASH‐P vs. DASH‐CF (p>0.05). These results indicate that substitution of lean pork for chicken and fish as the predominant source of protein does not influence the effectiveness of a DASH diet to improve blood pressure control.

Grant Funding Source: Supported by National Pork Board and NIH UL1 TR000006
Anti‐obese effect of ramalin in high‐fat diet‐induced obese mice (824.7)
Authors: Park Bongkyun , Lee Chunggi , Jang Yeonjeong , Pyo Suhkneung
Published: June 2021

The recent rise in obesity is tightly associated with an increase in diseases such as type 2 diabetes, cardiovascular disease and hepatic steatosis. Ramalin, isolated from the Antarctic lichen Ramalin terebrata, has been known to have various biological activities including anti‐inflammatory activity. However, other biological activities of ramalin remain unknown. We examined the anti‐obesity effect of ramalin in five week‐old mice that were fed either a control high‐fat diet (HFD), or HFD with oral administration 50 mg/kg or 100 mg/kg of ramalin for 8 weeks. Mice fed with ramalin showed 25% reduced body weight gain and ramalin treatment significantly inhibited epididymal fat‐pad weights compared with mice fed with HFD. Ramalin also reduced the levels of glucose, hepatic triglyceride, serum total, high‐density lipoprotein (HDL) and Low‐density lipoprotein (LDL) cholesterol. ELISA assay revealed that ramalin remarkably inhibited HFD‐induced leptin cytokine level. Furthermore, Real time PCR assay showed that HFD‐induced adipose tissue genes, known as adipogenesis markers, were significantly suppressed by ramalin. Overall, these results suggest that ramalin might be a plausible candidate to control obesity and obesity‐related disorders.
Anti‐inflammatory effects of JUVO (American style Saengshik) on high fat diet and dextran sulfate sodium‐induced colitis in C57BL/6J mice (824.8)
Authors: Park Kun‐Young , Song Jia‐Le , Shin Sung Ho , Park Myoung Gyu , Hwang Sung Joo
Published: June 2021

The aim of this study was to investigate the anti‐inflammatory effect of JUVO (J; an American style saengshik) on high fat diet(HFD, 45%) and dextran sulfate sodium (DSS, 2% wt/wt) induced ulcerative colitis (UC) in C57BL/6J mice. Saengshik (S) was used as a positive control. Body weight changes, colon length and ratio of colon length/weight were used to evaluate the preventive effects of JUVO in the colitis mice. Histological changes were observed by hematoxylin and eosin (H&E) staining. JUVO significantly suppressed HFD/DSS‐induced body weight losses and shortening of colon length, and decreased the ratio of colon length/weight. Histological observation suggested that JUVO suppressed edema, mucosal damage, and the loss of crypts induced by DSS. In addition, JUVO also decreased the serum and mRNA levels in the colon tissues of pro‐inflammatory cytokines including tumor necrosis factor‐α (TNF‐α), interleukin(IL)‐1β, IL‐6 and IL‐12. These results suggest that JUVO have an anti‐inflammatory effect on HFD/DSS‐induced ulcerative colitis in the mice, which through reduced pro‐inflammatory cytokine levels in serum and the colon tissue of the mice.
Soy germ for the prevention of prostate carcinogenesis in TRAMP mice (824.9)
Authors: Conlon Lauren , Erdman John W.
Published: June 2021

Prostate cancer (PCa) is the most commonly diagnosed cancer and second leading cause of cancer deaths in United States men. Soy germ has higher levels of daidzein and glycitein, and lower amounts of genistein compared to typical soy products. Genistein has been well studied, but less is known about daidzein and its microbially‐produced metabolite, (‐) equol, in PCa development. The objective of this research is to investigate the effect of feeding 2% soy germ, daidzein, or (‐) equol on the progression of PCa in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model. 3‐week old male C57BL/6 X FVB TRAMP mice were weaned from our breeding colony and immediately acclimated to a custom AIN‐93G control diet for one week. At 4 weeks of age, mice (n=30 per diet group) were randomized to one of four study diets, AIN‐93G control, AIN‐93G + 2% soy germ, AIN‐93G + 92 ppm daidzein, or AIN‐93G + 88 ppm (‐) equol until 18 weeks of age. Levels of daidzein and (‐) equol were matched to daidzein equivalents found in a 2% soy germ diet. Preliminary data shows no differences in food intake or body weight between the groups. Further analyses to be completed include prostate pathology scores, serum inflammatory markers, and epigenetic changes in prostate tissue. At the conclusion of this study, we will determine if a previously observed protective effect of soy germ is associated with the daidzein component or the daidzein metabolite, (‐) equol.

Grant Funding Source: Supported by NIH P50 AT006268
Correlation between dietary intake and plasma levels of choline and betaine in children with autism (827.1)
Authors: Hamlin JoAnna , Crook Tina , James Jill , Gonzales Dana , Hakkak Reza
Published: June 2021

Alterations in nutrient metabolism may exist in children with autism spectrum disorders (ASD’s), especially the pathways involving folate‐dependent one‐carbon metabolism. Few studies have investigated intake and plasma levels of choline and betaine in children with ASD, although they play a role in these metabolic pathways. This study examined 1) differences in plasma levels of choline and betaine in children with and without ASD and 2) relationships between dietary intake and plasma levels of choline and betaine in children with ASD. Secondary data analyses on 63 children were conducted. Significantly lower choline levels were found in children with ASD (M=8.00, SD=2.42) compared to children without ASD, M=10.03, SD=2.84; t (61) =‐3.04, p= 0.004, (two‐tailed). Similarly, betaine levels were significantly lower in children with ASD (M=24.1, SD=6.49) compared to children without ASD, M=36.70, SD= 6.73; t (60)= ‐7.5, p < 0.001, (two‐tailed). Strong, positive correlations were found between dietary intake and plasma levels of choline (r= 0.657, n=33, p<0.0005) and betaine (r=0.688, n=33, p<0.0005) in children with ASD. These results suggest that children with ASD may have diets inadequate in choline.

Grant Funding Source: HRSA; NICHD and Rochester University
Dietary sources of vitamin B12 intake among participants of the Adventist Health Study‐2 calibration study (827.14)
Authors: Damayanti Didit , Haddad Ella , Jaceldo‐Siegl Karen , Beeson Larry , Oda Keiji
Published: June 2021

Objective: To describe the contribution of dietary supplements, fortified foods, whole foods, and nutritional yeast to vitamin B12 intake among participants of the AHS‐2 calibration study.

Methods: We used cross‐sectional data from the AHS‐2 calibration study whose participants are representative sample of the AHS‐2 cohort. It included 66.7% females, 54.4% Whites, and have a wide variation of dietary intake (8.0% vegans, 28.2% lacto‐ovo‐vegetarians, 9.9% pesco‐vegetarians, 5.6% semi‐vegetarians to 48.3% non‐vegetarians). Dietary intake was assessed from six 24‐hour dietary recalls and entered into Nutrition Data System for Research (University of Minnesota). Dietary sources of vitamin B12 were classified as fortified foods, supplements, whole foods, or nutritional yeast. We identified food group ID 127 as supplements, foods containing vitamin B12 supplement with ID 21018 in ingredient level as fortified foods, foods containing yeast alone with ID 1913 as yeast and foods without fortification as whole foods.

Results: Sources of vitamin B12 intake among 922 participants of AHS‐2 calibration study.

Sources of vitamin B12
N
Mean ± SD
5th
Median
95th
Minimum
Maximum

Fortified foods (mcg)
922
3.24 ± 14.75
.00
1.50
7.54
.00
380.82

Supplements (mcg)
905
48.78 ± 177.57
.00
1.60
296.00
.00
2,142.86

Whole foods (mcg)
922
1.99 ±1.99
.09
1.41
6.14
.00
16.79

Yeast (mcg)
922
.01 ± .07
.00
.00
.01
.00
1.93

Conclusions: The highest sources of vitamin B12 intake came from supplements, followed by fortified foods and whole foods.
Folate receptor alpha in human plasma: relationship to folate status and effect of supplementation in healthy reproductive aged females (827.17)
Authors: Stamm Rosemary , Gray Andrew , Green Timothy , Houghton Lisa
Published: June 2021

Little research has been conducted to determine the effect of chronic folic acid (FA) intake on folate receptors (FR). We aimed to determine if long‐term supplementation with FA (140 or 400 μg/d) changes expression of FRα in blood. Samples from a 40‐wk randomized placebo controlled trial in 144 reproductive aged females were used. Plasma FRα was determined by ELISA, plasma folate vitamer concentrations by LC‐MS/MS, and red blood cell (RBC) folate by microbiologic assay. The geometric mean (SD) FRα concentration was 12.4 (1.7) nmol/L at baseline and 13.2 (1.6) nmol/L at 40‐wks. There were no significant correlations between FRα and any blood folate indices at baseline, however, after 40‐wks, FRα was significantly correlated with RBC folate (Spearman’s r = 0.217, P = 0.021). In a multivariable model, FRα at baseline significantly predicted FRα at 40‐wks (Ratio 2.41, 95% CI 2.23 – 2.60, P < 0.001). While there was no overall effect of treatment (P = 0.124), exploratory subgroup analysis revealed a borderline association of 400 μg/d FA with FRα at 40‐wks (Ratio 1.10, 95% CI 1.00‐1.20, P = 0.043), yet no effect of 140 μg/d FA (Ratio 1.04, 95% CI 0.95‐1.13, P = 0.414). This suggests that FRα in plasma, which may reflect FRα from decomposed RBCs and neutrophils, may be slightly increased by long‐term FA supplementation. Larger studies are needed to determine effects of chronic FA exposure on FR expression, including in other tissues.

Grant Funding Source: Supported by University of Otago Research Grant
Vitamin D supplementation effects on the IGF system in men after acute exercise (828.15)
Authors: Darr Rachel , Baker Mark , Wilding Gregory , Savage Kathy , Raslawsky Amy , Rideout Todd , Browne Richard , Burton Harold , Horvath Peter
Published: June 2021

Conflicting data between the IGF system and exercise may be due to IGF binding proteins. Vit D insufficiency is related to muscle weakness and IGFBP3. To determine if a combination of Vit D and acute exercise can alter the IGF system, Vit D insufficient and deficient men (39.0±8.6yo with 25OH D 20.0±7.7ng/mL) did one hour of stretching (ST), aerobic (AB), and resistive (RT) exercises, before and after 28 d of 4000IU/d Vit D3 (D, n=6) or Placebo (P, n=7). ST, an attention/time control visit, alternated passive movements. AB was moderate intensity treadmill walking. RT rotated moderate strength 50% 1‐RM repetitions (15, 10) of squat, bench press, leg press, and lat pull down. Serum total IGF1 (TIGF1), IGFBP1 (BP1), IGFBP3 (BP3) were measured before (T1, fasting), immediately after (T2), and 2 h post (T3) exercise. After ST, BP3 was higher in the D group at T2 (2948, 2130 ng/mL; p<0.03) and T3 (3087, 2212; p<0.02). During RT, TIGF1 decreased in the P group from T1 to T3 (151.4, 107.3 ng/mL; p<0.05), while BP1 increased in the D group from T1 to T3 (26.5, 96.2 ng/mL; p<0.05). RT BP3 was higher at T1, T2, and T3 in the D group (2932.5, 2110.7; p<0.03), (3163.9, 2392.5; p<0.04), and (3355.3, 2353.1; p<0.01). In AB, BP3 was higher in the D group at T2 (3128.6, 2226.3.0; p<0.04) and T3 (2949.7, 2135.1; p<0.05). D supplementation increased BP3 after low or moderate activity which may increase the delivery of IGF1 to tissues. Resistance exercise with Vit D not only heightened BP3 and BP1 levels but also maintained TIGF1 levels, potentially altering the IGF system for enhanced muscle health.