bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitamin D status is linked with plasma glutathione and cysteine thiol/disulfide redox status in adults (260.4)
Authors: Alvarez Jessica , Chowdhury Ritam , Jones Dean , Martin Greg , Brigham Kenneth , Binongo Jose , Ziegler Thomas , Tangpricha Vin
Published: June 2021

Objective: Redox status and inflammation are important in the pathophysiology of numerous chronic diseases. Epidemiological studies have linked vitamin D nutriture to a number of chronic diseases. We examined the relationships between serum 25‐hydroxyvitamin D (25(OH)D) concentrations and circulating biomarkers of redox and inflammation.

Methods: We studied a community‐based cohort of working adults in Atlanta, GA (N=693). Plasma thiol/disulfide redox status was determined by HPLC [glutathione (GSH), glutathione disulfide (GSSG), cysteine (Cys), cystine (CySS), and their calculated redox potentials (Eh), calculated by the Nernst equation]. Pro‐inflammatory biomarkers included IL‐6, IL‐8, TNF‐α, and C‐reactive protein. Relationships were assessed by multiple linear regression.

Results: Serum 25(OH)D levels were positively associated with plasma GSH levels, a more reducing GSH/GSSG EhCys, and inversely with Cys levels (P<0.001). These associations were not influenced by subject age, gender, race, % body fat, and cardiovascular risk factors (P<0.02). Serum 25(OH)D was not independently associated with any inflammatory marker.

Conclusion: Serum 25(OH)D levels were independently associated with major plasma thiol/disulfide redox systems, suggesting that vitamin D status may be involved in redox‐mediated pathophysiology.

Grant Funding Source: Supported by NIH (T32DK007298, UL1TR000454, K23AR054334, and K24RR023356).
SIRT1 dependent IRS1 phosphorylation mediates the effect of vitamin D on GLUT4 translocation and glucose utilization via PI3K/PTEN/PTP1B/ PIP3/AKT/PKCζ/λ signaling cascade in high glucose treated 3T3L1 adipocytes (260.5)
Authors: Manna Prasenjit , Jain Sushil
Published: June 2021

This study examined the biochemical mechanism by which VD regulates glucose homeostasis in diabetes. 3T3L1 adipocytes were treated with VD (1,25(OH)2D3, 0‐50 nM, 2 h) and high glucose (HG, 25mM, 20 h). Results (n=4) showed that VD upregulated SIRT1 protein expression and IR phosphorylation, stimulated IRS1/PI3K/PTEN/PTP1B/PIP3/AKT/PKCζ/λ signaling cascade, and increased GLUT4 translocation (44%), and glucose utilization in HG treated cells. VD also upregulated its receptor (VDR) protein expression (45%) and decreased ROS (31%), NOX4 protein expression (71%), and NF‐κB phosphorylation (47%) in HG treated cells. VD treatment along with insulin enhanced GLUT4 translocation (29%) and glucose utilization (32%) compared to either insulin or VD alone in HG treated cells. Signal silencing studies showed that in IR siRNA transfected cells, VD can increase the IRS1 phosphorylation, GLUT4 translocation, and glucose utilization; however the effect of VD was significantly inhibited in the SIRT1 siRNA transfected cells. Studies with VDR siRNA demonstrated that the effect of VD on glucose metabolism in HG treated cells is mediated via its receptor. This study demonstrates a novel molecular mechanism by which VD upregulates SIRT1 protein expression and IRS1 phosphorylation which mediate the GLUT4 translocation and glucose utilization via PI3K/PTEN/PTP1B/PIP3/AKT/PKCζ/λ signaling cascade in adipocytes.

Grant Funding Source: Supported by NIH RO1 AT007442 and Malcolm Feist Chair in Diabetes
γ‐Tocopherol attenuates liver injury in diabetic (db/db) mice with nonalcoholic fatty liver disease (260.7)
Authors: Mah Eunice , Li Youyou , Guo Yi , Lim Yunsook , Jalili Thunder , Symons J , Bruno Richard
Published: June 2021

Oxidative stress and inflammation during diabetes contribute to nonalcoholic fatty liver disease (NAFLD). We hypothesized that antioxidant and anti‐inflammatory activities of γ‐tocopherol (γ‐T) would protect against liver injury associated with NAFLD. Diabetic (db/db) mice were fed a diet containing γ‐T at 0 or 500 mg/kg and wild‐type mice (WT) were fed no γ‐T for 8 wk prior to examining parameters of NAFLD, oxidative damage, and inflammation. Db/db mice fed γ‐T had higher hepatic γ‐T (7.2±0.9 vs 2.8±0.3 pmol/mg) and γ‐carboxyethyl‐hydroxychroman (313±54 vs 15±0 pmol/mg) relative to db/db controls. Db/db mice had greater liver and adipose mass, serum lipids, and glucose relative to WT; these were unaffected by γ‐T. Histological scoring indicated that γ‐T did not affect hepatic steatosis, hepatocyte ballooning, or inflammatory infiltration observed in db/db mice. Hepatic malondialdehyde (MDA) and serum alanine aminotransferase (ALT) were higher in db/db mice compared to WT, but were normalized in db/db mice fed γ‐T. Serum ALT was correlated (P<0.05) to hepatic MDA (r = 0.50) and inversely correlated to hepatic γ‐T (r = ‐0.60). γ‐T lowered hepatic iNOS protein expression that was otherwise increased in db/db mice. Collectively, greater hepatic γ‐T status protects against liver injury during NAFLD, likely by reducing lipid peroxidation and inflammation, but without affecting liver steatosis or hyperglycemia.
Prohibitin1 regulates H19 and insulin‐like growth factor 2 expression in hepatocytes and liver (261.1)
Authors: Ko Kwang Suk , Yang Heping , Li Tony , Lee Sang Hoon , Song Young Sun , Lu Shelly
Published: June 2021

BACKGROUND&AIMS: Prohibitin1 (Phb1) is a ubiquitously expressed and highly conserved protein that acts as a mitochondrial chaperone and possibly as a tumor suppressor in liver. We reported liver‐specific Phb1 knockout (KO) mice develop multifocal hepatocellular carcinoma (HCC) and knockdown of Phb1 in AML12 cells (a normal hepatocyte cell line) accelerated growth. H19 and insulin‐like growth factor 2 (Igf2) are two imprinted genes often overexpressed in HCC that are induced in the Phb1 KO livers. Here we examined how Phb1 regulates their expression. METHODS: Real‐time PCR and Western blotting measured expression in Phb1 KO livers and AML12 cells treated with various siRNAs. RESULTS: H19 RNA and Igf2 mRNA and protein levels are tremendously upregulated with Phb1 deficiency in vivo (by several hundred folds) and in vitro (2‐fold after 24 hour Phb1 siRNA treatment). E2F1 and c‐Myc can activate H19 transcriptionally, but knockdown or either c‐Myc or E2F1 did not block H19 or Igf2 induction with Phb1 knockdown. Ctcf, an insulator protein that represses Igf2 expression when it is bound to the imprinting control region, is downregulated with Phb1 knockdown in AML12 cells. Double knockdown of Phb1 and Ctcf resulted in the same magnitude of H19 and Igf2 induction as single knockdown of either Phb1 or Ctcf, suggesting Phb1 may regulate the expression of H19 and Igf2 through Ctcf. Interestingly Ctcf expression is lower while H19 and IGF2 are induced in HCC samples as compared to normal liver. CONCLUSIONS: Loss of Phb1 lowers Ctcf expression, which in turn allows H19 and Igf2 gene induction to occur.

Grant Funding Source: Supported by NIH R01 (R01CA172086) and NRF Korea Fund (2012R1A1A1012261)
The role of sirtuin 3 in the differential pro‐oxidant effects of (‐)‐epigallocatechin‐3‐gallate in oral cells (261.4)
Authors: Tao Ling , Lambert Joshua
Published: June 2021

Dietary approaches may play an important role in reducing oral cancer burden. The green tea catechin, (‐)‐epigallocatechin‐3‐gallate (EGCG), has been reported to have cancer preventive effects. Previously, we found that EGCG induced mitochondrial dysfunction, oxidative stress and apoptosis in oral cancer cells but exerted antioxidant effects in normal oral cells. In this study, we aim to investigate the role of sirtuin 3 (SIRT3), an important mitochondrial redox balance regulator, in EGCG‐induced differential pro‐oxidant effects in oral cells. We found 100 µM EGCG significantly decreased SIRT3 mRNA (15%‐56%; 1‐12 h) and protein levels (17%‐34%; 6‐24 h), as well as, activity (58%; 12 h) in SCC‐25 oral squamous carcinoma cells. By comparison, there was no significant effect of EGCG on SIRT3 mRNA and protein levels, but a remarkable increase of SIRT3 activity (7.5‐fold; 12 h) in HGF‐1 human gingival fibroblast cells. The estrogen‐related receptor alpha (ERRα) regulates transcription of SIRT3. EGCG significantly decreased the mRNA (20%‐44%; 3‐6 h) and protein levels (31%‐50%; 6‐24 h) of ERRα in SCC‐25 but not HGF‐1 cells, indicating that EGCG regulates SIRT3 expression through modulation of ERRα. The effect of EGCG on transcription activity of ERRα is under investigation. This study identified SIRT3 as a novel target of EGCG, thus providing additional mechanistic information for future in vivo studies.

Grant Funding Source: Supported by a grant from the American Institute for Cancer Research (Grant #10A102 to JDL)
Needs assessment for childhood obesity intervention program (262.1)
Authors: Harper Kristin , Berenson Danielle , Calkin Katie , Buffalo Julie , Mullis Rebecca , Anderson Alex
Published: June 2021

The purpose of this study was to assess the needs of families of obese children to inform the development of a clinic‐based program to manage the weight and health of obese children. This cross‐sectional study assessed selected nutritional and behavioral habits and the family environment of obese children who were seen in a private pediatric practice. Primary caregivers of 20 obese children were interviewed via phone using a structured questionnaire containing closed‐ and open‐ended questions. Descriptive statistics were used for data analysis with results reported as percentages and means. The mean age of the children was 9.2±2.9 yrs. The majority of the children (55%) were males. Seventy‐five percent of the parents/guardians spoke Spanish as the primary language. Mean fruit and vegetable intake was 3.2±1.9 servings/d. Three‐quarters of children were reported to consume sugar‐sweetened beverages, with a mean intake of 1.9±1.6 cups/d. Although caregivers reported their children played actively for an average of 54.3 min/d, the majority of the children spent over 2 hrs on screen time. About 90% of parents/caregivers reported that they are currently working on changing their children’s dietary and health habits. Our preliminary results suggest a need for clinic‐based program with emphasis on improving nutritional and behavioral habits of obese children and their families while ensuring it is culturally relevant.

Grant Funding Source: Supported by: The William B. Mulherin Foundation for Health & Wellness
Maternal vitamin D status during pregnancy is associated with infant anthropometry during the first year of life (267.1)
Authors: Eckhardt Cara , Gernand Alison , Roth Daniel , Bodnar Lisa
Published: June 2021

Maternal vitamin D deficiency during pregnancy has been linked to fetal growth and may also impact child growth in infancy. We used data and blood samples from the 12‐site U.S. Collaborative Perinatal Project (1959‐65) to examine the association of maternal vitamin D status during pregnancy and infant anthropometry across the first year of life in singletons (n=2473 mother‐child pairs). Maternal serum 25‐hydroxyvitamin D (25(OH)D) was measured at 蠄26 weeks gestation using chromatography‐tandem mass spectrometry. Infant anthropometry was expressed as z‐scores for length (LAZ), head circumference (HC‐Z), weight (WAZ), and BMI (BMI‐Z) based on measures at birth and 4, 8 and 12 months of age. We controlled for study site, and important infant and maternal characteristics in linear mixed effects models. 25(OH)D <30 nmol/L (conventional definition of deficiency) vs. 蠅30 nmol/L was associated with LAZ and HC‐Z measures that were 0.13 (95% CI: 0.03‐0.23) and 0.20 (95% CI: 0.11‐0.28) lower, respectively, across the first year of life. Differences observed by 25(OH)D in WAZ and BMI‐Z at birth resolved by 4 months of age, potentially reflecting catch‐up growth. These findings suggest that maternal vitamin D deficiency has a sustained association with markers of linear or skeletal growth during the first year of life, while initial associations with measures of weight and body composition may resolve.

Grant Funding Source: Supported by R01 HD 056999
Diet patterns and clustering of cardiometabolic risk during adolescence (267.5)
Authors: Moore Lynn , Singer Martha , Bradlee M. , Daniels Stephen
Published: June 2021

Objective: To examine the relation between adolescent dietary patterns and cardiometabolic risk (CMR) clustering at end of adolescence.

Methods: Data for 1369 girls enrolled at 9‐10 years of age in the longitudinal NHLBI Growth and Health Study were used. Girls with complete data over 10 years on diet, and six CMR factors, and all confounders of interest were included. Mean dietary data were derived from 3‐day records. CMR factors analyzed included increased waist circumference, elevated blood pressure, insulin resistance, low HDL‐cholesterol and high LDL‐cholesterol and triglyceride levels. Multiple logistic regression analysis was used to estimate the relative risk of having multiple (蠅 2 or 蠅 3) risk factors in late adolescence associated with usual dietary patterns from 9‐17 years of age.

Results: Of 1369 subjects included, 252 (18.4%) had 3‐6 risk factors while 479 (35.0%) had two or more. Higher intakes of fruit and non‐starchy vegetables, dairy, and grains were independently associated with having fewer risk factors as were eating patterns characterized by higher combined intakes of these food groups. After adjusting for age, race, socio‐economic status, height, physical activity, and television watching, girls with high intakes of both dairy and fruits and non‐starchy vegetables (vs. lower intakes) were nearly 50% less likely to have 蠅 3 CMR factors while girls with higher intakes of grains and fruits and non‐starchy vegetables (vs. lower intakes) were nearly 60% less likely to have 蠅 3 CMR factors.

Conclusion: These data suggest that CMR clustering at the end of adolescence is associated with dietary patterns during adolescence.
Dietary cholesterol and heart health: a systematic review and meta‐analysis (267.6)
Authors: Berger Samantha , Raman Gowri , Vishwanathan Rohini , Jacques Paul , Johnson Elizabeth
Published: June 2021

Objectives: To conduct a systematic review and meta‐analysis evaluating the effects of dietary cholesterol on cardiovascular outcomes in healthy adults.

Methods: Medline, Cochrane Central, and CAB databases were searched through March 31, 2013 for prospective studies in English‐language that quantified dietary cholesterol and cardiovascular or lipid outcomes in adults. Random effect models meta‐analysis was used when at least 3 studies reported the same outcome.

Results: 28 studies (16 cohorts with 361,923 subjects and 9 randomized trials with 579 subjects) published between 1981 and 2013 were eligible. Dietary cholesterol was not significantly associated with ischemic heart disease deaths (4 cohorts, summary relative risk [RR] 1.35, 95% CI 0.91, 2.01) or ischemic stroke (4 cohorts, summary RR 1.13, 95% CI 0.99, 1.28). Dietary cholesterol intake 蠅300mg/day compared to <300 mg/day, had a small but statistically significant increase in plasma total cholesterol (9 trials, net change 6.76 mg/dL, 95% CI 0.71, 12.80, p=0.029) and LDL cholesterol (9 trials, net change 5.41 mg/dL, 95% CI 0.62, 10.20, p=0.027), but no significant change in HDL cholesterol.

Conclusions: Reviewed studies were heterogeneous and lacked methodological rigor. Carefully controlled efficacy studies and well‐conducted cohort studies may still be useful to pin down the relative effects of dietary cholesterol on cardiovascular outcomes.

Grant Funding Source: This study is supported by the Egg Nutrition Center
Food security and household food expenditure in Guatemala (268.4)
Authors: Puente‐Duran E. Violeta , Melgar‐Quinonez Hugo
Published: June 2021

In Guatemala malnutrition and food insecurity are highly prevalent. Distinguishing between food security levels can help improve understanding of dietary patterns among specific groups, thus allowing for more effective and specialized designs of food assistance programs. The Escala Latinoamericana y Caribena de Seguridad Alimentaria (ELCSA) is a validated tool that distinguishes food secure households from three food insecurity levels: mild, moderate and severe. The purpose of this study was to assess expenditure on food by food security level in 13,482 Guatemalan households in the 2011 Living Conditions Survey (ENCOVI), in order to identify potential nutritional gaps in the given groups. Information from the 115 items survey section “Expenses and Consumption of Food” and the food security section containing ELCSA was used. Analyses were made regarding food items purchased during the twelve months previous to the survey. One way ANOVA was used to compare mean differences among food security categories on expenditures made for various food groups. Bonferroni post‐hoc analyses were used for multiple comparisons between groups with a confidence level set at 95%. Food secure households had significantly higher expenditures on meats, dairy, fruits, and vegetables when compared to food insecure groups. Severely food insecure households had a higher expenditure on staple grains and sugar, and the proportion of expenditure was significantly higher for starches, sweets, and non‐meat proteins (eggs and beans). As food insecurity level increases, expenditure on boneless beef decreases, while expenditures on chicken interiors increase. Different food consumption by food insecurity level indicates the need for differentiated interventions and programs.
Longitudinal changes on nutritional disorders, nutrient intake and household food insecurity in children ongoing surgery to correct cleft lip and palate (268.8)
Authors: Wall‐Medrano Abraham , Martínez‐Ruiz Nina , Ramos‐Jiménez Arnulfo , Rodríguez‐Tadeo Alejandra , Astiazarán‐García Humberto
Published: June 2021

Introduction. In Mexico, public health care programs aimed to improve the quality of life of children with cleft lip (CL) and/or palate (CP) are scarce. Instead, humanitarian aid programs like “Thousand Smiles” are gaining recognition nationwide. Objective. To document the longitudinal changes on the prevalence of nutritional disorders, nutrient intake inadequacies and household food insecurity (HFI) among pediatric patients with CL&P participants of “Thousand Smiles” campaigns 2009‐2011. Methods. 123, 101 and 76 (25 being 100% program adherents) patients [CL (13%), CP (26%) and CL&P (61%), mean age 2.4, 68% male] were recruited at stage 1 (before 1st surgery), 2 (before 2nd surgery) and 3 (蠅three weeks after 2nd surgery). Participant´s height & weight, two 24h food recalls, and a socioeconomic & HFI questionnaire, were obtained from each participant per stage. Comparisons across stages were performed for overall stage participants and in those 100% adherents. Results. At stage 1, 20%, 24%, 5% and 5% of all patients showed low weight, stunting, wasting and overweight/obesity respectively, <10% were exclusively breastfed, 35%, 17%, 13% and 7% had iron, vitamin A, vitamin C and calcium inadequacies (蠄75 RDI) and 55% of all households experienced moderate‐to‐severe HFI (HFI‐MS). The prevalence of stunting (p=0.002), inadequacies for iron & vitamin A (p<0.01) and calcium & vitamin C (p<0.05) and HFI‐MS (p=0.002) fell from the 1st to the 3rd stage. Conclusion. “Thousand Smiles” campaigns aimed to correct CL&P of pediatric patients form northwestern Mexico; modify their prevalence of malnutrition, nutrient inadequacy and even household food insecurity.
Nicotinic acid synergizes with leucine to stimulate AMPK/Sirt1 signaling and regulates lipid accumulation and lifespan in C. elegans (269.7)
Authors: Bruckbauer Antje , Zemel Michael
Published: June 2021

Previous data demonstrate the BCAA leucine (Leu) to stimulate AMPK and Sirt1 signaling in vitro and in vivo. Nicotinic acid (NA) has been used as a highly effective treatment of dislipidemias; however, its poorly tolerated side effects (severe vasodilation and flushing at high doses) limits its use as a drug. Since it increases NAD+ and thereby activates Sirt1, we tested the synergistic effects of leucine (0.5 mM) and low dose NA (10 nM) on AMPK/Sirt1 signaling and downstream effects in cell culture and C. elegans. The NA‐Leucine combination increased P‐AMPK by 80%, P‐AMPK/AMPK ratio by 114% (p<0.001) and Sirt1 by 25% (p=0.04) in adipocytes and myotubes, while the individual components exerted no independent effect. In C. elegans, NA‐Leu increased P‐AMPK and DAF‐16 (FOXO) signaling by 66% (p=0.018) and 105% (p=0.004), respectively. This was associated with a reduction in lipid accumulation by 49% (p=0.017) and a 28% increase in median survival (p=0.0001) under mild oxidative stress conditions (0.2 mM paraquat). These data indicate that low concentrations of NA synergize with leucine to modulate sirtuin signaling and lipid metabolism, and may therefore be a potential therapeutic for treatment of metabolic diseases.

Grant Funding Source: NuSirt Sciences Inc.
Chronic flavan‐3‐ol exposure induces changes in metabolism and transport kinetics in differentiated Caco‐2 cell monolayers (270.1)
Authors: Redan Benjamin , Ferruzzi Mario
Published: June 2021

A limiting factor in the application of flavan‐3‐ols for chronic disease prevention is their low oral bioavailability. However, evidence suggests that chronic dosing paradigms can significantly increase their bioavailability. To explore fundamental alterations occurring from chronic gut exposure to flavan‐3‐ols, Caco‐2 monolayers were cultured and differentiated in 6‐well plates in addition to Transwell® inserts for 12 and 21 days, respectively. Culture media contained 0, 1, or 10 μM concentrations of epigallocatechin gallate (EGCG) or epicatechin (EC). Following differentiation, cellular uptake and transport of flavan‐3‐ols was assessed from 0‐6 h following a final 50 μM acute treatment of respective pretreated compound. Transport of flavan‐3‐ols, along with metabolism to methylated and sulfonated metabolites, was assessed by LC‐TOF‐MS. After 90 min, 1 μM EGCG pretreated monolayers showed significantly (P < 0.05) greater accumulation compared to control (mean ± SEM; 310. ± 12.4 vs. 208 ± 12.1 pmol EGCG/mg protein). 1 μM EC pretreatment showed similar significant (P < 0.05) results (477 ± 65.9 vs. 150. ± 35.0 pmol EC/mg protein). Further, Papp calculated from Transwell® experiments for 10 μM pretreated cell monolayer were 7.93 x 10‐7 cm/s compared to 5.09 x 10‐7 cm/s for control. These results suggest that intestinal transport and metabolism of flavan‐3‐ols is modulated by chronic exposure.

Grant Funding Source: Supported by National Science Foundation Graduate Research Fellowship Program Grant No. DGE‐1333468.
Pharmacokinetics and brain distribution of polyphenols from fruit extracts in young pigs (270.2)
Authors: Janle Elsa , Chen Tzu‐Ying , Kritchevsky Janice , Hargett Katherine , Feller Kathryn , Klobusnik Ryan , Tabaczyk Olivia , Ferruzzi Mario , Jouni Zeina
Published: June 2021

To assess the bioavailability and brain accretion of dietary polyphenols, pigs (14 ± .5 lb) were divided into 5 groups (n=8/group) of control (water), low (27.5 mg/kg) or high (82.5 mg/kg) concentration of bilberry extract or apple/grape seed extract (A/GSE). Polyphenol ingredients were delivered once a day. After 3 weeks, a plasma 24h pharmacokinetic study was conducted. Blood and brain regions (stem, cerebellum, cortex, hypothalamus, hippocampus and amygdala) were harvested 24 h later and analyzed for key polyphenol metabolites from (epi)catechin, quercetin and anthocyanins by HPLC‐MS/MS. Polyphenols were not detected in control plasma or brain samples. A significant (p <.05) plasma dose response was observed for epicatechin and catechin metabolites in the A/GSE groups. Most of the plasma anthocyanin metabolites in bilberry groups exhibited significant or trend (0.05 < p < 0.1) dose responses. Brain accumulation varied (in pmol/g tissue): (epi)catechin (7‐1430), quercetin (0.1‐1.2), anthocyanins (0‐3). Significant differences were observed between different brain regions and not all metabolites were detected in all brain regions. The highest concentration for most metabolites was in the cerebellum in both high concentration groups. These data demonstrate that polyphenol metabolites can cross the blood‐brain barrier and accumulate at different concentrations in different brain regions.

Grant Funding Source: Supported by Mead Johnson Nutrition
Ellagic acid attenuates adipocyte differentiation via histone arginine methylation‐associated epigenetic modification (271.2)
Authors: Kang Inhae , Okla Meshail , Chung Soonkyu
Published: June 2021

Previously, we have demonstrated that ellagic acid (EA), a naturally occurring polyphenol in numerous fruits and vegetables, represses adipogenic conversion of human adipose‐derived stem cells (hASCs) by altering epigenetic factors. However, the detailed mechanism by which EA attenuates adipocyte differentiation is largely unknown. In this study, we first hypothesized that upregulation of HDAC9 by EA is the key mechanism to inhibit adipogenesis. Unexpectedly, knockdown HDAC9 by siRNA had marginal impact on recovering adipogenesis. Intriguingly, EA treatment was associated with reduced histone 3 arginine 17 methylation levels (H3R17me2), implying the inhibitory role of EA in CARM1 (coactivator associated arginine methyltransferase 1) activity. To test the hypothesis that recovery of H3R17me2 will rescue EA‐mediated inhibition of adipogenesis, cell‐penetrating peptides of CARM1 (CPP‐CARM1) was delivered to compensate the loss of CARM1 activity by EA. Delivery of CPP‐CARM1 was able to restore H3R17me2, HDAC9 dissociation, histone acetylation levels, PPARγ expression, and triglyceride accumulation. Taken together, our data suggest that reduced CARM1 activity by EA results in a decrease of H3R17me2 levels, which may interrupt consecutive histone remodeling steps for adipocyte differentiation including histone acetylation and HDAC9 dissociation from chromatin.

Grant Funding Source: Supported by USDA‐HATCH
Folate in a high multivitamin gestational diet causes obesogenic phenotype and epigenetic alterations in hypothalamic feeding pathways in Wistar rat offspring (271.3)
Authors: Cho Clara E. , Pannia Emanuela , Huot Pedro S.P. , Sánchez‐Hernández Diana , Kubant Ruslan , Bazinet Richard P. , Anderson G. Harvey
Published: June 2021

High multivitamin (HV, 10‐fold AIN‐93G) gestational diets fed to Wistar rats produce offspring with higher food intake, obesity and characteristics of metabolic syndrome. The objective was to determine if folate within the HV diet contributes to obesogenic phenotype through epigenetic effects on hypothalamic regulatory mechanisms. Male offspring of dams fed HV diet with normal folate (NF) (HVNF) were compared with those from HV fed dams. Offspring were weaned to a high fat diet for 8 weeks. HVNF offspring had lower food intake (5%, P=0.0005), body weight (4%, P=0.03), fat preference (12%, P=0.02), adiposity index (10%, P=0.04), plasma leptin concentration (24%, P=0.03), glucose response to a glucose load (18%, P<0.05) and insulin resistance (12%, P=0.03). They had higher pro‐opiomelanocortin (POMC; 38%, P<0.05) and lower leptin receptor (Ob‐Rb; 10%, P=0.03), mammalian target of rapamycin (MTOR; 7%, P=0.01), serotonin receptor (5‐HTR) 1A (11%, P=0.03) and 5‐HTR2A (21%, P=0.03) gene expression. DNA methylation of 5‐HTR1A (P=0.0007) was lower but higher in 5‐HTR2A (P=0.01). Phosphoinositide 3‐kinase (PI3K) signaling protein expression was improved (42%, P=0.03). In conclusion, folate in the HV gestational diet contributes to the obesogenic phenotype in the offspring concurrent with alterations in gene expression, DNA methylation and energy sensing in the hypothalamus that favor increased food intake.

Grant Funding Source: Supported by Canadian Institutes of Health Research, Institute of Nutrition, Metabolism and Diabetes
Quantity of dietary protein, but not pattern of intake, affects net protein accretion in older adults (272.4)
Authors: Kim Il‐Young , Schutzler Scott , Wolfe Robert , Ferrando Arny
Published: June 2021

We examined 16‐h whole body protein turnover [protein synthesis (PS), breakdown (PB), and net balance (NB), expressed as delta changes from the fasted state] in four permutations: a level of protein intake of 0.8g/kg/day (1RDA) or 1.5g/kg/day (2RDA) with an uneven [U: 15/20/65%)] or even distribution patterns of intake (E: 33/33/33%) for breakfast, lunch, & dinner of intake over the day. Twenty healthy older subjects were randomly assigned into one of 4 groups: 1RDA‐U, 1RDA‐E, 2RDA‐U, or 2RDA‐E, and were studied with primed continuous infusions of ring‐[2H5]phenylalanine & ring‐[2H2]tyrosine on day 4 following 3 days of diet habituation. Positive NB was achieved at both protein levels, but NB was significantly greater in 2RDA vs. 1RDA [NB: 94.75 ± 6.01 vs. 58.86 ± 4.91 g protein/kg/750min; p = 0.0001], without effects of distribution on NB. The greater NB was due to the higher PS with 2RDA vs. 1RDA [PS: 15.40 ± 4.84 vs. ‐17.98 ± 8.44 g protein/kg/750min; p = 0.0018]. PS was significantly correlated with delta changes of mean plasma concentrations of leucine from the fasted state (R2 = 0.51, p = 0.0004). We conclude that in the context of mixed meals whole body net protein accretion is greater with protein intake above recommended dietary allowance (0.8 g protein/kg/day), regardless of protein intake pattern, primarily through increased protein synthesis.

Grant Funding Source: Supported by Dairy Research Institute and Award Number UL1TR000039 and KL2TR000063 (NCATS and NIH)
The addition of non‐fat dairy products to the regular diet reduces central blood pressure and improves vascular function (272.5)
Authors: Machin Daniel , Park Wonil , Alkatan Mohammed , Mouton Melissa , Tanaka Hirofumi
Published: June 2021

Elevated central systolic blood pressure and vascular dysfunction increase the risk of cardiovascular disease. Using a randomized, crossover study design, we tested the hypothesis that the addition of non‐fat dairy products to the regular diet would reduce carotid systolic blood pressure (cSBP) and improve vascular function. Forty‐four adults with elevated brachial blood pressure (52±2 yr, 133±1/80±1 mmHg) underwent a high dairy condition (+4 servings/day non‐fat dairy products) and isocaloric non‐dairy condition (+4 servings/day fruit products) in which all dairy products were removed. Both dietary conditions lasted four weeks with a 2‐week washout before crossing over. After the high dairy condition, cSBP decreased, brachial flow‐mediated dilation (FMD) increased, and carotid‐femoral pulse wave velocity (cfPWV) decreased (‐3±1mmHg, +1.1±0.5%, ‐66±18 cm/sec, respectively; p<0.05). After the non‐dairy condition, cSBP or cfPWV did not change, but FMD was reduced (‐1.0±0.5%, p<0.05). ∆cSBP was positively related to ∆cfPWV (r=0.59). In conclusion, the addition of non‐fat dairy products to the regular diet reduced central systolic blood pressure and improved vascular function. Additionally, the hypotensive effects of non‐fat dairy products may in part be mediated by reductions in central arterial stiffness.

Grant Funding Source: Supported by Dairy Research Institute
It takes a village: community collaboration to promote consumption of dry beans in school meals (273.2)
Authors: Riddle Lee Anne , Betz Drew , Atterberry Kelly , Rueda Janice , Miles Carol
Published: June 2021

Pulse crops, which include dry beans, dry peas, lentils and chickpeas, are nutrient dense foods rich in fiber, protein, iron, potassium and phytonutrients. Consumption of pulses is associated with increased satiety and reduced risk of chronic diseases, yet these low‐cost foods are not readily used in school food service operations. A unique collaboration of community partners aims to promote increased consumption of locally produced pulses among K‐12 students in Washington. In this pilot study, one elementary school (61% free & reduced eligible) and one high school (45% free & reduced eligible) were selected for the study, and 66 students were recruited to participate. Extension specialists from Agriculture and Nutrition developed four pulse‐based recipes. Food Service directors chose one pulse‐based recipe to prepare at their school and plate waste studies were conducted to assess students’ consumption. Elementary students had reduced pulse consumption while high school students had an increase in consumption. Results suggest development of recipes in closer collaboration with school food service directors could be an effective strategy to increase consumption of locally produced pulse crops in school food programs.
Potentiation of TNF‐induced inflammatory transcriptional regulation by SFK activation (278.3)
Authors: Adam Alejandro , Lu Yao Wei , Alsaffar Hiba , Martino Nina , Lowery Anthony , Schwarz John , Vincent Peter
Published: June 2021

TNF induces dramatic changes in the transcriptional profile of endothelial cells to promote a pro‐inflammatory phenotype that allows leukocyte transendothelial migration and loss of barrier function. We have previously shown that treatment with 50 pg/ml TNF increases endothelial permeability after 4‐6 h in monolayers of human dermal microvascular endothelial cells expressing DN‐Csk to activate endogenous Src Family Kinases (SFKs) but not in control (LacZ) monolayers. We thus reasoned that this delay in increasing permeability could be due to a requirement in transcriptional changes. Here, we demonstrate that activation of SFKs potentiates the transcriptional effects induced by TNF. After 6 h, TNF elevated ICAM1, E‐Selectin, HSPA1A/B and complement C3 mRNA levels 41‐, 76‐, 12‐ and 24‐fold respectively. In the presence of DN‐Csk, the same treatment induced these genes 149‐, 310‐, 38‐ and 202‐fold, respectively. Similar effects were observed in the mRNA of TNF itself (46 vs 193 fold) and lymphotoxin A (5 vs 21 fold), but not lymphotoxin B (86 vs 83 fold). ATF3 was induced 7 and 20‐fold respectively. EGR2 mRNA was induced 5‐fold by DN‐Csk and 9‐fold by the dual treatment, while Fos mRNA was induced 4‐fold in DN‐Csk‐expressing cells after 6h TNF and 9‐fold after 18 h. While these treatments did not lead to changes in the expression of many junctional genes, ZO‐1 and VE‐cadherin localization was drastically altered as determined by immunofluorescence. A pro‐inflammatory transcriptional profile, together with local posttranslational changes at the cell‐cell junctions, could explain the synergistic loss of barrier function observed with dual TNF signaling and endogenous SFK activation.

Grant Funding Source: Supported by an AHA SDG grant 13SDG17100110 to APA.
The role of female pelvic width on walking economy and speed: Benefits with few tradeoffs (335.2)
Authors: Wall‐Scheffler Cara , Myers Marcie
Published: June 2021

Among human sexually dimorphic features, a few are particularly important for locomotion, namely: overall body size, the proportions and lengths of the lower limbs, and the shape of the pelvic girdle. Across populations, males are absolutely larger in terms of mass, stature and limb length, whereas females have wider pelves relative to size. Variation between the sexes in these traits has consequences for the energetic cost of transport (CoT) and the speed of travel and as such should result in differences including sex‐specific energetic costs and strategies of mobility. Here, data are presented on the energetic differences between males and females (N=39) walking with and without loads. Without loads, females have significantly slower optimal walking speeds (10.9%, p=0.019) but have lower costs at their optimum (23.4%, p=0.002), and a broader base to their optimal walking curve (thus more available walking speeds) (22.1%, p=0.276). Furthermore, people with relatively wider bi‐trochanteric breadths (for their mass; generally females) have even lower costs at their optimal speeds (p=0.036). During loaded walking, a relatively wider pelvis increases both stride length (p<0.001) and speed flexibility (p<0.001), providing a morphological offset for load‐related costs. These data suggest that females’ wide pelves provide energetic and speed‐related benefits during both loaded and unloaded walking.
Locomotor adaptations and the origin of the hominin pelvis (335.4)
Authors: Ward Carol
Published: June 2021

The bony pelvis is involved in many aspects of an animal’s biology, and what factors shaped the unique form of the human pelvis has received a great deal of attention in paleoanthropology. Not only does the bony pelvis surround the birth canal, but it anchors lower limb muscles and reflects the shape of the bony torso. Torso shape and lower limb function are integrally related to locomotor adaptation, and so either or both can affect obstetric adaptations. Because extant great apes share a similar pattern of pelvic morphology with broad, coronally oriented iliac blades and long lower pelvis, for many years it was reasonable to hypothesize that this represented the ancestral condition for hominins, and that the transition to habitual terrestrial bipedal locomotion involved substantial selection to reorient the ilia and shorten the pelvis, and thus restrict the bony birth canal. However, recent fossil evidence from Miocene and early Pliocene hominoids, notably Rudapithecus hungaricus (9 ma, Rudabanya, Hungary) and Ardipithecus ramidus (4.4 Ma, Aramis, Ethiopia) suggest that the pattern of selection that shaped the hominin pelvis may not have involved a drastic reduction in iliac length and reorientation, but that other factors such as spinal mobility and hip joint orientation played a much greater role than previously thought in influencing the size and shape of the birth canal in early hominin evolution.
Big anatomy: he new views from integrated anatomy, development, and genes (338.6)
Authors: Mabee Paula
Published: June 2021

Comparative anatomical data are increasingly sought after to guide computational genomics work, to develop evo‐devo hypotheses, and to inform interpretation of organismal evolution in light of molecular phylogenies. Anatomical data, however, are embedded in the literature and uncomputable, thus impairing the ability of investigators to find, aggregate, or link them to other data types. By using ontology‐based methods to represent anatomical features, the Phenoscape team has developed a knowledgebase where the morphological variation of vertebrates, with a specific focus on for fin and limb phenotypes, is linked to genes and phenotypes of zebrafish, Xenopus, mouse and human through their community databases. Using the reasoning enabled by the ontology structure, and assuming a high‐level of conservation of developmental processes, candidate genes can be automatically inferred for morphological features that characterize clades and species. Two such predictions were tested in the wet‐lab, and the results support the broad‐scale utility of this approach. Morphological data in this computable format can be browsed, sorted and aggregated in ways that present unprecedented possibilities for data mining and discovery. Current work by the team includes development of methods for rapid retrieval of similar phenotypes and a focus on the fin/limb skeletal transition.
Validation of a unilateral heating model to increase hindlimb length in growing mice (342.4)
Authors: Tamski Holly , Stanko Laura , Godby Justin , Efaw Morgan , Schlierf Thomas , Salisbury Travis , Ion Gabriella , Serrat Maria
Published: June 2021

Limb length inequality is prevalent in nearly 90% of the population. Differences of 20mm can cause gait asymmetry and chronic back pain. Surgical treatments are invasive and alternatives are needed. Exposure to warm ambient temperature accelerates limb elongation in mice, suggesting that targeted heat could be a noninvasive way to promote limb length equalization. We tested the hypothesis that daily heat exposure on one side of the body will unilaterally increase limb length. Female mice (N=6) were treated with 40C unilateral heat for 40 minutes per day for 14 days post‐weaning. Controls (N=6) were not treated. Femur length was increased 1.3% on the heat‐treated side. Tibial elongation rate, measured by oxytetracycline labeling, was increased >12%. Body mass was unaffected. Histological analysis revealed morphological differences in the growth plate. To validate a protein assay for the next step, we developed western blotting methods to asses IGF‐I activation in whole proximal tibias. We show that IGF‐I (100ng/ml) stimulates Akt phosphorylation after 90 minutes in culture, suggesting that this method will be effective for measuring IGF‐I signaling in heat‐treated bones. This model will be important for elucidating mechanisms of heat‐enhanced growth. Our findings are relevant for the development of non‐invasive heat‐based therapies to increase length of specific bones without surgery.

Grant Funding Source: Supported by NASA WV Space Grant Consortium and UK CCTS (NIH UL1TR000117)
Therapeutic delivery of placental stem cells to modulate vasculature and promote fracture repair (342.6)
Authors: Bahney Chelsea , Taylor Aaron , Sadat Ali , Tormos Kathryn , Miclau Ted , Maltepe Emin , Marcucio Ralph
Published: June 2021

Blood supply to a bone fracture is a critical determinant of the rate and extent of healing. Therapies designed promote bone healing by stimulating angiogenesis have been proposed for a long time, yet, to date no effective treatments are available. In this work, we capitalize on a transient process that modifies the vasculature during pregnancy to support the developing fetus. Placental progenitors, trophoblast stem cells (TSCs), promote vasculogenesis in response to fetal hypoxia by physically remodeling the maternal arterioles and secreting angiogenic factors to generate a high volume, low pressure fluid exchange. We hypothesize that the therapeutic application of TSCs will promote vascular remodeling and enhance fracture healing. Our data shows that TSCs injected to non‐stabilized murine fractures engraft into the vasculature and enhance the local blood supply. Furthermore, injection of TSCs increased the volume of the cartilage callus 7‐days post fracture, leading to more bone after 14‐days of healing. To our knowledge, this is the first study evaluating the therapeutic potential of trophoblasts. Our results have the potential to enhance clinical outcomes in skeletal trauma, where there is often poor vascular perfusion. Importantly, this work may also have a significant impact on the broader field of regenerative medicine, since loss of organ function is often intimately tied to compromised vascularity.

Grant Funding Source: NIH ‐ NRSA F32
Ultrasound imaging in anatomy: a supplement to dissection (343.3)
Authors: Lee Vaughan , Schneider Brandt
Published: June 2021

Advances in ultrasound technology have made ultrasound imaging an important skill for physicians. To prepare our students for this clinical skill, TTUHSC/SOM is incorporating ultrasound in the undergraduate medical curriculum. Ultrasound imaging naturally integrates with anatomical dissections, providing a method to view anatomy in a clinical manner. Ultrasound sessions were designed for first year students and integrated with dissection sessions of the wrist, neck and liver. Wrist and neck sessions were conducted in a classroom with 15 ultrasound machines and 30 students in pairs scanning each other. The liver session was conducted in a simulation center with simulated patients and small groups of five students. The exercises were assessed with pre‐ and post‐test for each session. Average grades increased from pre‐ to post‐test; wrist (57.0/85.8), neck (75.8/92.8), and liver (71.4/95.4). Student opinions were assessed by survey on a five‐point Likert scale. Improved understanding of anatomy received scores of 3.43, 4.13 and 4.19, respectively, for the three sessions. Students indicated a desire for more in‐depth ultrasound sessions, including such statements as “LOVE the ultrasound sessions!” Supplementing anatomy sessions with ultrasound imaging enhanced dissections and introduced students to clinical skills.

Grant Funding Source: Supported by the TTUHSC/SOM
A comparative examination of odontogenic markers in both toothed and toothless amniotes (344.3)
Authors: Lainoff Alexis , Moustakas Jacqueline , Kallonen Aki , Hämälainen Keijo , Smith Kathleen , Hu Diane , Marcucio Ralph , Hlusko Leslea
Published: June 2021

It is a well‐known tenet of murine tooth development that BMP4 and FGF8 antagonistically initiate odontogenesis, but it is largely unexplored whether this tenet is conserved across amniotes. Additionally, changes in BMP4‐signaling have previously been implicated in evolutionary tooth loss in Aves. Here we demonstrate that Bmp4, Msx1, and Msx2 expression is limited proximally in the red‐eared slider turtle (Trachemys scripta) mandible, at stages equivalent to those at which tooth development is initiated in mice, similar to previously reported results in chicks. To address the question of whether the limited domains shared between the turtle and the chicken are indicative of an evolutionary molecular parallelism that resulted in the loss of the dentition, or whether the domains simply constitute an ancestral phenotype, we assessed expression of Bmp4, Msx1, Msx2, and Fgf8 expression in a toothed reptile (the American alligator, Alligator mississippiensis) and a toothed non‐placental mammal (the Gray short‐tailed opossum, Monodelphis domestica). We demonstrate that the Bmp4 domain is limited proximally in M. domestica and that the Fgf8 domain is limited distally in A. mississippiensis just preceding odontogenesis. Additionally, we show that Msx1 and Msx2 expression patterns in these species differ from those found in mice. Our data suggest that a limited Bmp4 domain does not necessarily correlate with edentulism, and reveal that the initiation of odontogenesis in non‐murine amniotes may be more complex than previously imagined. Our data also suggest a partially conserved odontogenic program in T. scripta, as indicated by the conserved mandibular expression domains of Pitx2, Pax9, and Barx1 and by the presence of Shh‐expressing palatal epithelium in some Yntema stage 17 (Y17) T. scripta specimens, which we hypothesize may represent potential dental rudiments based on the fossil record of Testudinata.

Grant Funding Source: Supported by NIH‐NIDCR‐R01DE018234 and NSF‐BSC‐0616308 with REU supplement.
Activation of musculoskeletal development and repair mechanisms in the regenerating lizard tail (344.7)
Authors: Hutchins Elizabeth , Tokuyama Minami , Eckalbar Walter , Fisher Rebecca , Rawls Alan , Wilson‐Rawls Jeanne , Kusumi Kenro
Published: June 2021

Uniquely among amniote vertebrates, lizards can lose their tails and regrow a functional replacement. These regenerated tails, which have an organization distinct from the original tail, contain newly formed hyaline cartilage, muscle, vasculature, spinal cord, and skin. Using Illumina RNA‐Seq technology, we have identified over 300 differentially expressed (DE) genes along the proximal/distal axis of the regenerating tail at a mid‐growth stage (n=5). Histological and immunohistological analysis reflect these transcriptomic data. Specifically, there are proliferative cells throughout the proximal/distal axis and proliferative markers are expressed throughout the proximal/distal axis. However, cells progressively show differentiation towards the proximal region of the regenerating tail, and genes with high expression in this region include myogenic regulators and factors for muscle structure and differentiation. In contrast, genes with high expression in the regenerating tail tip include factors for wound and inflammatory response, hormonal and homeostatic regulation, and skeletal system development, including regulation of the Wnt pathway. Thus, these studies have identified components of a genetic program for regeneration in the lizard that includes both developmental and adult repair mechanisms, indicating value in the translation of these findings to future regenerative therapies.

Grant Funding Source: Supported by NIH NCRR/ORIP Grant R21RR031305 and Arizona Biomedical Research Commission