bluuebunny/crossref_metadata_2025 huggingface dataset:
NF‐kappaB activation in the fetal lung promotes macrophage maturation (715.3)
Authors: Stouch Ashley , Zaynagetdinov Rinat , Barham Whitney , Stinnett Amanda , Yull Fiona , Hoffman Hal , Blackwell Timothy , Prince Lawrence
Published: June 2021

In preterm infants, exposure to inflammation increases the risk of chronic, developmental lung disease bronchopulmonary dysplasia. While macrophages are the key cells that initiate lung inflammation, little is known about lung macrophage phenotype and maturation. We hypothesized that fetal lung macrophages mature into distinct subpopulations during mouse development, and that activation could influence macrophage maturation. Expression of the fetal macrophage markers CD68, CD86, CD206, Ym1, fibrinogen‐like protein 2 (FGL2), and indolamine‐2, 3‐dioxygenase (Ido1) was developmentally regulated. Flow cytometry analysis measured a narrow spectrum of CD11b expression in fetal lung macrophages, with separation into CD11bloF4/80hi and CD11bhiF4/80lo subpopulations in newborn and adult lung. Similar to adult alveolar macrophages, fetal lung macrophages responded to the TLR4 agonist LPS and the alternative activation cytokines IL‐4 and IL‐13. Using a macrophage‐specific constitutively active IKKβeta transgenic model (IKFM), we demonstrated that macrophage activation increased proinflammatory gene expression and reduced the response of fetal lung macrophages to IL‐4 and IL‐13. Fetal IKFM lungs contained increased percentages of more mature, CD11bloF4/80hi cells that also expressed higher levels of the alternative activation markers CD204 and CD206. The maturational programming of fetal lung macrophages into mature alveolar macrophages therefore did not follow traditional proinflammatory and alternative activation paradigms.

Grant Funding Source: Supported by National Institutes of Health Grants HL097195, HL086324, HL116358, HL069765
Apical volume expansion results in an immediate ENaC activation but in delayed onset of water resorption (718.1)
Authors: Wittekindt Oliver , Schmidt Hanna , Michel Christiane , Neubauer Daniel , Frick Manfred , Mizaikoff Boris , Dietl Paul
Published: June 2021

Apical volume expansion results in an activation of the epithelial sodium channel ENaC. Its activation is proposed to facilitate water resorption and thus to counteract increases of apical surface liquid (ASL) volume e.g. in lung edema. In this study we investigated the adaptation of epithelial water resorption to apical volume expansion (AVE) in NCI‐H441 epithelia. Cells were cultivated at air‐liquid interface. AVE was introduced by adding isotonic NaCl solution onto the apical surface. Water resorption was measured by the recently established deuterium oxide dilution method (Neubauer et al. 2013, Anal Chem). ENaC channel activation was quantified in Ussing chamber experiments as amiloride sensitive short circuit currents. As expected, ENaC activity increases to its maximum within 15 min after AVE. Water resorption does not follow ENaC activation but starts 4 to 6 h after AVE. When ASL volume is reduced to 3.7 ±0.2 μl / 0.33 cm^2 water resorption stops and ENaC activity is reduced to basal levels. Our data demonstrate that AVE results in a rapid ENaC upregulation but ENaC activation is not the only trigger of onset of water resorption. However, off set of water resorption is associated with ENaC down regulation.

Supported by BIU D.5007

Grant Funding Source: Supported by BIU.5007
The ubiquitylated protein profile of rat cortical neurons is altered in a primary culture model of stroke (728.12)
Authors: Chua Michael Dominic , Pasqualotto Bryce , Rintoul Gordon , Guttman Julian
Published: June 2021

Ischemic stroke is initiated by the lack of oxygen and glucose that are normally involved in maintaining the ion gradients at the presynaptic terminals. Overstimulation of postsynaptic glutamate receptors induces a massive influx of calcium, mitochondrial depolarization, and accumulation of reactive oxygen species causing neuronal death. To begin to understand the cellular effects on neurons following a stroke, we hypothesized that protein ubiquitylation would be altered due to its crucial role in cell signalling and protein labeling for subsequent degradation. To test this hypothesis, we analyzed immunoblots of glutamate‐treated primary rat cortical neuron cultures. We found that several low‐molecular weight proteins in the glutamate‐treated neuron lysates showed increased ubiquitylation levels as compared to untreated neuron lysates. The altered ubiquitylation profile of only a subset of proteins suggests that select small proteins are tightly regulated during stroke. Through this study, we have identified proteins specifically targeted during stroke. Their identification opens the door to targets for therapeutic intervention to rescue neurons following ischemic brain injury.

Grant Funding Source: This study was funded through Departmental Funds
Anorexigenic effect of serotonin in an alternative vertebrate model (728.18)
Authors: Zhang Wei , Didehvar Dillon , Cline Mark , Gilbert Elizabeth
Published: June 2021

Serotonin (5HT) is a central anorexia factor that inhibits food intake in mammals. Here, we used an avian model to study the effect of 5HT on appetite regulation, which may provide insight on the evolutionary aspects of food intake. Chicks treated with 24 nmol 5HT had reduced food intake up to 30 min following injection. Those that received 48 nmol 5HT had reduced food intake following injection throughout the 60 min observation time. The dose of 12 nmol was not effective. No chick treated with 5HT reduced water intake. We designed experiment 2 to determine if the anorexigenic effect of 5HT was primary by conducting a comprehensive behavior analysis. Those chicks treated with 5HT had increased deep rest, decreased feeding pecks, locomotion, steps, defecations, and chirps, but time spent sitting, standing, and preening were not affected. Exploratory pecks, jump, and drinks were also not affected. In sum, 5HT is associated with sedation, however, the effects on food intake are likely not due to malice because 5HT chicks did not reduce water intake and the number of drinks. These data support that 5HT function as a direct satiety factor in chicks.
Effects of essential fatty acids associated with low‐power pulsed ultrasound on skin injury healing: experimental comparative analysis (734.3)
Authors: Crisci Ana Rosa , dos Santos Poliana , da Silva Fabiana , Jorge Maria Helena
Published: June 2021

Effects of essential fatty acids associated with low‐power pulsed ultrasound on skin injury healing: Experimental comparative analysis Ana Rosa Crisci1, Poliana Branco dos Santos1, Fabiana Colombari da Silva1, Maria Helena Simões Jorge1 1Department of Morphology, Universitarian Center Barao de Maua, Ribeirao Preto, Sao Paulo This study aimed to assess the benefits of essential fatty acids (EFA) in low cost traditional topic treatment association with low‐power pulsed ultrasound (USP). The USP is a non invasive treatment and have been confirmed efficacy in acceleration of the healing process of several kinds of tissues. Fifteen male rats were divided into four groups: 1 was treaty with EFA, 2 was treaty with USP, 3 was treaty with EFA and USP and 4 had just the injury cleanse with saline solution 0,9%. The rats were treaty unlimited access to water and food before and after the surgical procedure. The animals were anaesthetized with ketamine chloridrate 100mg/Kg and xylazine chloridrate 6mg/Kg. Trichotomy follow by a subcutaneous stinging lesion (1cm2) was done in animals and the same received daily treatment without anesthesia. The animals were sacrificed in the fifth day after injury. For histology analysis, skin samples were collected and quantified in accordance with Myers modified. Statistical analysis was made 舃舃a nonparametric Kruskal ‐ Wallis test. The data obtained by histopathological analyzes showed that the healing process was more efficient with the use of EFA after 5 days of treatment. The average scores of the lesions treated with the three groups have not statistically significant difference ( p < 0.02 ). The results of this study show that the USP was more effective in the healing of skin wounds in rats in the first 5 days.
Regulation of the mammalian target of rapamycin signaling by grape polyphenols in breast cancer (766.6)
Authors: Rivera Rivera Amilcar , Castillo Pichardo Linnette , Dharmawardhane Suranganie
Published: June 2021

The Akt/adenosine monophosphate protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway has emerged as a critical signaling nexus for regulating cellular metabolism, energy homeostasis, and cell growth; and dysregulation of this pathway contributes to the development of metabolic disorders such as obesity, type 2 diabetes, and cancer. We previously reported that a combination of grape polyphenols, (resveratrol, quercetin and catechin: RQC) at equimolar concentrations, inhibits breast cancer (BC) growth and metastasis in nude mice. The current objective is to investigate a role for RQC in BC therapy via inhibition of the Akt/AMPK/mTOR pathway. We found that in metastatic BC cells, RQC at 5uM each inhibits Akt and mTOR activities and activates AMPK, an endogenous inhibitor of mTOR. To determine the contribution of individual polyphenols to the effect of combined RQC on mTOR signaling, metastatic BC cells were treated with RQC individually and in combination, at various concentrations. The activities (phosphorylation) of AMPK, Akt, and the mTOR downstream effectors p70S6 kinase (p70S6K) and 4E binding protein (4EBP1) were determined by Western blot. Results show that at 蠅5uM, individual compounds downregulated Akt activity to a level similar to combined RQC at 5uM each, indicating a single mechanism of action. Quercetin at 15uM was the most efficient compound at both AMPK activation and mTOR inhibition. Therefore, regulation of cancer cell metabolism by quercetin has the potential to block BC cell growth and migration, and thus, metastatic progression. In conclusion, quercetin is a viable candidate for future development as an anti BC therapeutic.

Grant Funding Source: Supported by US Army DoD/BCRP W81XWH‐07‐1‐0330 to SD, U54 CA096297 to UPRM‐MSC, MBRS‐RISE
Calcium Intake modulates obesity phenotypes in African American children possessing risk alleles for development of type 2 diabetes (767.11)
Authors: Devaney Joseph , Sprouse Courtney , Gordish‐Dressman Heather , Ryan Leticia , Tosi Laura
Published: June 2021

Objective: Explore whether total calcium consumption modulates obesity phenotypes in African American children (AA) possessing risk alleles for development of type 2 diabetes (T2D).

Methods: 142 AA children (5 to 9 yrs) were genotyped for alleles associated with increased risk of developing T2D. 67 single nucleotide polymorphisms (SNPs) were chosen from published GWAS to identify T2D risk loci. We evaluated three obesity phenotypes; body mass index percentile and, using DXA, percent fat, and total body fat/height. Assessments of nutritional intake (milk, dairy, and total calcium) were measured using a validated food frequency questionnaire, and then evaluated to assess their effect on the significant outcome/SNP relationships. All analyses used linear or quantile regression models.

Results: We found significant relationships between body fat outcomes and 5 SNPs (ADAMTS9‐AS2, CDC123, PRC1, ZMIZ1, and rs7560163). Assessment of the effect of nutritional intake showed that total calcium intake normalized for body weight was a highly significant predictor of body fat and a significant modifier of the outcome/SNP effect.

Conclusions: In AA children, calcium intake modulates obesity phenotypes in individuals possessing risk alleles associated with development of T2D. Expansion of this research may help identify individuals for whom increased calcium intake will have the most beneficial effect.

Grant Funding Source: Dairy Research Institute
A new method for measurement of ACE2 activity in tissues using fluorescent angiotensin peptides (779.9)
Authors: Oliveira Marilene , Santos Robson
Published: June 2021

The carboxypeptidase ACE2 is a key enzyme of the renin‐angiotensin system, being primarily responsible for the formation of Ang ‐(1‐7 ) by cleavage of the carboxy‐terminal end of Ang II as well as the degradation of Angiotensin A to alamandine. The aim of this study was to develop a new method for measurement of ACE2 activity using the fluorecent Ang A and Ang II and HPLC. To evaluate the activity of ACE2, we used samples of heart, kidney and lung of FVBN wild type mice. Samples were homogenized in buffer for optimal activity of ACE2. Initially, we optimized the conditions of the assay using different concentrations of recombinant human enzyme rhACE2 at different incubation times with the substrate Ang II or Ang A. After the assay optimization, our results showed a linear formation of Ang‐(1‐7) or Alamandine linear with the concentration of RhACE2 and time of incubation. The homogenates of the tissues and the cocktail of inhibitors used for the incubation did not produce fluorescence. With the addition of the protease inhibitors Z ‐prolyl‐ prolinal , Captopril Chymostatin, Phosphoramidon , DL Thiorphan , Amastatin , Bestatin , Benzilsuccinaeo and PCMB, it was possible to inhibit the enzymes related to the SRA except ACE2 ,ensuring the reproducibility of the assay. Losartan also used to prevent the binding of the substrates to AT1 receptors present in homogenates. The ACE2 inhibitor MLN4760,abolished the formation of Ang‐(1‐7) or Alamandine by the homogenates. However, as described previously, DX600 was not able to inhibit ACE2 in mice tissue homogenate. The sensitivity of the assay was higher with the use of Ang A as substrate. This novel non‐isotopic, HPLC‐based method can be used for studies on the functional role of ACE2 in physiological and pathological conditions.

Grant Funding Source: INCT‐CNPq, CAPES, FAPEMIG
Epigenetic regulation of EGF‐induced CTEN gene expression (800.2)
Authors: Liao Yi‐chun , Chang Chia‐chun
Published: June 2021

CTEN induced by epidermal growth factor (EGF) mediates the EGF‐promoted cell migration and also prolongs EGF receptor (EGFR) signaling. However, the molecular mechanism underlying EGFR signaling upregulates CTEN expression is still incompletely understood. In this report, we showed that EGF‐induced expression of CTEN was mediated by ERK activation in a normal human prostatic epithelial cell line RWPE‐1 as well as in a cervical cancer cell line HeLa. Further dissection of the regulatory mechanism at transcriptional and posttranscriptional levels found that serial 5’‐deletions within 3‐kb CTEN promoter produced no significant induction in promoter activities after EGF stimulation. In addition, EGF has no effect on CTEN mRNA stability. Nonetheless, by chromatin immunoprecipitation assay, we revealed that EGF treatment enhanced the histone acetylation levels in the promoter region of CTEN. Furthermore, anacardic acid, a histone acetyltransferase p300 inhibitor, suppressed EGF‐induced CTEN expression, suggesting that the histone acetyltransferase p300 may be involved in the regulation of EGF‐induced CTEN expression.

Grant Funding Source: Supported by National Science Council, the Republic of China (Taiwan)
Preconception anemia and birth outcomes in Vietnam (804.6)
Authors: Gonzalez‐Casanova Ines , Nguyen Phuong , Hao Wei , Pham Hoa , Truong Truong , Nguyen Son , Martorell Reynaldo , Ramakrishnan Usha
Published: June 2021

The negative impact of anemia during pregnancy on birth outcomes is known, but few have examined the relative contribution of anemia prior to conception. We analyzed data on preconception anemia (PA) and birth outcomes collected prospectively from 946 mother‐infant pairs participating in a micronutrient supplementation trial in Vietnam. We used generalized linear models to estimate associations between PA, defined as a three‐level ordinal variable (coded 0,1,2) based on hemoglobin concentrations (Hb): >12 mg/dl (80% of sample), >11 to <12 mg/dl (13%), and <11 mg/dl (7%), and to adjust for potential confounders (BMI, age, education). We found negative associations between PA and birth weight (‐47 g, p=0.03), length (‐0.41 cm, p=0.02), head circumference (‐0.32 cm, p=0.03), and gestational age‐GA (‐0.29 weeks, p=0.01). After controlling for GA, the associations between PA and measures of birth size were attenuated (~0.00, p>0.10), suggesting that GA mediated the association between PA and birth size in our study population. These findings support the need for strategies and interventions to prevent and control anemia in women of reproductive age, both prior and during pregnancy, to improve birth outcomes.

Supported by The Micronutrient Initiative and The Mathile Institute for the Advancement of Human Nutrition
Food insecurity is associated with health related quality of life in HIV infected adults (805.13)
Authors: Hatsu Irene , Hade Erinn , Campa Adriana
Published: June 2021

Objective: HIV disease affects Quality of life (QOL), making it an important outcome in disease management and treatment. QOL can also be affected by food insecurity. We therefore investigated the relationship between food security status and health related quality of life of persons living with HIV (PLHIV).

Methods: After consenting, 176 individuals completed questionnaires that assessed sociodemographic variables, and health related quality of life (using SF‐36 V2). Analyses for associations were conducted by either Fisher’s exact test or general linear models.

Results: Participants were 41% food secure (FS), 34% low food security (LFS) and 25% very low food security (VLFS). LFS was associated with lower scores for vitality (p=0.01), social functioning (p=0.01), and mental health (p=0.02). VLFS was associated with lower scores for role‐physical (p=0.01), general health (p<0.001), vitality (p=0.001), social function (p=0.001), role‐ emotional (p=0.003), and mental health (p<0.001). Both LFS and VLFS were associated with higher bodily pain (p=0.03 and p=0.03 respectively). They were also significantly associated with reduced scores within the mental component summary scale (MCS); p=0.03 and p<0.001 respectively.

Conclusion: VLFS is strongly associated with physical and mental wellbeing of PLHIV. Strategies to improve food access and subsequent wellbeing in this population are needed.
Ethnic differences in pregnancy outcome and its predictors in Kuwait (806.9)
Authors: Rahman Abdur , Al‐Rashidi Haila , Akhtar Saeed
Published: June 2021

Background: Kuwait is a multi‐ethnic society with two‐third of the population comprises expatriates of various nationalities. In this research, we evaluated the major predictors of pregnancy outcome, and tested whether there are ethnic differences in maternal variables, and infant related birth attributes.

Methods: Women (n=369) admitted to the Al‐Sabah Maternity Hospital Kuwait for deliveries were recruited in this study. Data were collected on mother height (MH), hemoglobin (Hb), mid‐upper arm circumference, blood pressure, placental weight (PW), birth weight (BW), crown‐heal length (CHL), and head circumference (HC). PW/BW ratio was also calculated.

Results: Linear regression analysis showed PW as a significant predictor of BW, CHL and HC and MH as a significant predictor of BW and CHL. Both PW and MH were significantly lower in Kuwaiti and Indian women compared to Egyptian, Syrian or Lebanese women. Parallel to these, Kuwaiti and Indian women had babies with significantly lower HC and CHL values. Hb was significantly lower in Kuwaiti women compared to other ethnic groups, whereas, BW did not differ significantly.

Conclusion: Significant ethnic differences were observed in CHL and HC and these differences were predicted by PW and MH. Further research is needed to elucidate the significance of these differences in pregnancy outcome and factors that contribute to these differences.

Grant Funding Source: Supported by KU Graduate Student Grant
A family‐centered lifestyle intervention focused on milk and alternatives reduces adiposity in 6‐ to 8‐ year‐old overweight and obese children compared to control: results at 6 months from a RCT (808.24)
Authors: Cohen Tamara , Hazell Tom , Loiselle Sarah‐Eve , Kasvis Popi , Vanstone Catherine , Kim Nicolas , Plourde Hugues , Rodd Celia , Weiler Hope
Published: June 2021

The association between adiposity and milk and alternative (milk and alt.) intakes in prepubescent children is unclear. This study assessed the impact of a family‐based lifestyle intervention on adiposity in overweight (n= 14) and obese (n=64) children (age 7.5 ± 0.78 y). Children were randomized to control (CTRL) [no intervention], standard (SInt) [2 milk and alt./d] or intensive (IInt) [4 milk and alt./d] interventions focused on meeting the Canadian food and activity guides. Measurements were height (Ht; m) and weight (Wt; kg) to compute body mass index for‐age‐ z‐scores (BAZ) using WHOAntroPlus Software (WHO 2007), waist circumference (WC; cm) and percentage body fat (%BF) assessed using dual‐energy X‐ray absorptiometry. Using mixed model ANOVAs, no differences were seen at baseline among groups for BAZ or %BF. At 6‐months, Ht increased compared to baseline (p=0.898) and was not different among groups. Compared to baseline, %BF (‐2.4±0.92%; p=0.0015) and BAZ (‐0.5±0.08; p<0.001) was reduced only in IInt whereas WC increased only in CTRL (+2.5±1.61 cm; p=0.007). These preliminary results demonstrate the short‐term benefits of participating in a lifestyle intervention focused on increased milk and alt and activity guidelines in the context of the family environment. Study funded by the Dairy Farmers of Canada Research Cluster Initiative (Agriculture and Agri‐Food Canada, Dairy Farmers of Canada and the Canadian Dairy Commission).

Grant Funding Source: Dairy Farmers of Canada Research Cluster Initiative
Consumption of sugar‐sweetened and non‐sugar‐sweetened beverages of urban Mexican children (810.26)
Authors: Rodriguez‐Ramirez Sonia , Gaona‐Pineda Elsa , Cuevas‐Nasu Lucia , Shamah‐Levy Teresa
Published: June 2021

Objective: To estimate the consumption of sugar‐sweetened and non‐sugar‐sweetened beverages (SSBs and Non‐SSBs) of pre‐school and school‐age children in urban Mexico.

Methods: This analysis included pre‐school (n=211) and school‐age children (n=115), from the study “Food consumption in Mexican population”, which is a cross‐sectional, multi‐stage, stratified study, representative of the urban area in Mexico. Dietary data were collected using three multiple‐pass 24‐h recalls, in non‐consecutive days. The beverage consumption was categorized in: 1) SSBs with non‐benefit (regular soft drinks, sweetened coffee and tea, sweetened beverages), 2) SSBs with some benefit (milk‐based beverage), 3) Non‐SSBs (diet soft drinks, unsweetened coffee and tea, and unsweetened milk), and 4) Plain water. The average of the three days of volume (ml) and energy (kcal) was obtained. Results are weighted by design.

Results: The consumption in pre‐school‐age children was: 83 ±14.2 ml/d of regular soft drinks; 27±10.2 ml/d of sweetened coffee and tea; 216±21 ml/d of other sweetened beverages; 120±17 ml/d of sweetened milk; 19±5 ml/d diet soft drinks; 194±16 ml/d of unsweetened milk; and 335±35.4 ml/d of plaint water. In school‐age children the consumption was: 153±16.4 ml/d of regular soft drinks; 26±7.6 ml/d of sweetened coffee and tea; 215±24 ml/d of other sweetened beverages; 119±18 ml/d of sweetened milk; 22±6.6 ml/d diet soft drinks; 80±12 ml/d of unsweetened milk; and 460±43.3 ml/d of plaint water. The energy from SSB in both groups is approximately 15% of the total energy.

Conclusion: SSBs consumption is high in the children diet. It is important to identify the principal types of this beverage to establish intervention to improve the diet.

Grant Funding Source: Nestlé Mexico
Moderate alcohol consumption and hemoglobin A1C in women with previous gestational diabetes (811.6)
Authors: Ferranti Erin , Dunbar Sandra , McCullough Marjorie , Ziegler Thomas , Narayan K. M. Venkat , Reilly Carolyn , Foster Jennifer
Published: June 2021

Objective: We examined alcohol consumption in relation to hemoglobin A1C levels in women within five years of a pregnancy affected by gestational diabetes (GDM).

Methods: This study was a cross‐sectional design. Habitual alcohol consumption was assessed with the Block 2005 food frequency questionnaire. Elevated hemoglobin A1C levels were defined as 蠅 5.7% and measured with the Bayer A1C Now+ meter. Body mass index (BMI) was calculated from measured height and weight. Data analysis included descriptive, bivariate statistics, and multiple linear regression.

Results: Participants (n = 74, 54% non‐white); mean age 35.6 (SD = 5.5) years and were 2.5 (SD = 1.6) years since their GDM delivery. Nearly half (47.3%) had an elevated A1C. Mean alcohol intake was 0.44 (SD = .64) servings/day. Greater consumption was associated with lower A1C (rho = ‐.45, p = <.0001). Alcohol intake was higher in those without diabetes risk (M = .59, SD = .66 servings per day) compared to those with diabetes risk (M = .27, SD = .58 servings/day; t = 2.2, p = .03). In multiple regression analysis, adjusted for age/race and BMI, alcohol intake was not associated with A1C (β = ‐.14, p = .21), but BMI was (β = .48, p = <.0001).

Conclusions: Women within five years of a GDM pregnancy are at high risk for T2DM development. Although the unadjusted analysis revealed that alcohol had a protective effect on A1C, BMI was the only significant independent predictor of glycemia in this small sample. Clinical efforts focused on weight management should be a high priority to reduce T2DM risk.

Grant Funding Source: Supported by the American Heart Association, National Institute for Nursing Research, NIH,
Effect of olive leaf extract on the gene expression for lipid metabolism in ovariectomized rats (814.6)
Authors: Yoon Leena , Liu Yanan , Kim Hyun‐Sook
Published: June 2021

There have not been enough studies regarding estrogen deficiency and lipid metabolism. Even though many postmenopausal women gain excess adipose tissues, the actual mechanism has not been yet fully understood. Thus, we measured mRNA expression after oral administration of olive leaf extract, which has been reported as an antioxidant, hepatoprotective, and hypolipidemic agent, in ovariectomized rats. Ovariectomized rats were administered either 200 or 400 mg/kg body weight of olive leaf extract (n=7 for each group) every day for 10 weeks. Expression of peroxisome proliferator‐activated receptor alpha and acyl‐CoA oxidase mRNA in the liver were significantly higher in the rats administered lower and higher dose of olive leaf extract compared with ovariectomized control. However, mRNA expression of liver carnitine palmitoyltransferase1a was not different among the groups. Expression of peroxisome proliferator activated receptor gamma mRNA in adipose tissue was decreased in the rats with 400 mg/kg body weight of olive leaf extract than ovariectomized control but it was not significantly different among the groups. Adipose triglyceride lipase expression and lipoprotein lipase mRNA did not change compared to ovariectomized control. These results suggest that genes involved in lipid metabolism may be regulated by olive leaf extract administration in ovariectomized rats. Transcription factors for further analyses to expand current results might include sterol regulatory element binding protein 1c and acetyl CoA carboxylase.
Meta‐analysis of genome‐wide association studies for circulating phylloquinone (818.9)
Authors: Dashti Hassan , Shea M , Smith Caren , Tanaka Toshiko , Nalls Mike , Kritchevsky Stephen , Liu Yongmei , Ordovás José , Booth Sarah
Published: June 2021

Phylloquinone (PK) is the primary dietary and circulating form of vitamin K. Low circulating PK has been associated with abnormal calcification of soft tissue. Because few studies have assessed the genetic component influencing circulating PK, we sought to identify common genetic variants associated with circulating PK concentrations by conducting a genome‐wide association (GWA) meta‐analysis of circulating PK in two populations of European descent from the Cohorts of Heart and Aging Research in Genomic Epidemiology (CHARGE) Consortium. Results from adjusted cohort‐specific GWA analyses were meta‐analyzed with inverse variance weights. No genome‐wide significant associations were observed in the meta‐analysis (n = 2,138). We found nominal associations (P<1×10−6) on five independent loci. These results suggest that the APOA1C3/A4/A5 gene cluster, involved in triglyceride transport, CYP4F2, involved in vitamin metabolism, COL22A1, a collagen gene, CTNAA2, a catenin gene, and CDO1, a cysteine dioxygenase gene, are candidate genes that may be associated with serum PK. Given the multiple determinants of circulating PK and small sample size of the current study, further investigation with a larger sample is warranted.
Higher sodium and saturated fat intake is associated with lower muscle protein synthesis in elders (820.16)
Authors: Markofski Melissa , Timmerman Kyle , Dickinson Jared , Jennings Kristofer , Reidy Paul , Borack Michael , Rasmussen Blake , Volpi Elena
Published: June 2021

INTRODUCTION: The relationship between muscle protein synthesis (MPS) and habitual diet in older adults is not known.

OBJECTIVE: We examined which nutrients were the best predictors of basal MPS.

METHODS: We analyzed pre‐treatment data are from a larger, ongoing clinical trial of interventions for sarcopenia. Older males and females (n=42; 71.5±1.8y) recorded habitual food intake for three days preceding an afternoon admission to the CRC. Subjects were fed a standardized dinner and 10PM snack, after which were only allowed water ad libitum. The next morning, MPS was measured by infusing 13C6Phenylalanine and collecting two muscle biopsies from the vastus lateralis, two hours apart. A stepwise linear regression was fitted with measures of dietary composition, aerobic fitness, and muscle strength and function.

RESULTS: The best model (R2=0.654, P=0.000197) predicting MPS included lean body mass and palmitoleic acid as positive predictors, and VO2peak, sodium, eicosapentaenoic acid, and stearic acid as inverse predictors. Protein intake was not a model component. Muscle strength and function were not a component of the model.

CONCLUSION: Sodium, saturated fat and aerobic fitness may negatively impact basal MPS in older adults. The lack of an effect of protein intake and the negative effect of VO2 peak will require further investigations.

Grant Funding Source: Supported by NIH/NIA R01 AG030070 and NIH/NIA R01 AG030070S1
Characterization of muscle‐specific branched‐chain alpha‐ketoacid dehydrogenase kinase (BDK)‐deficient mice (820.6)
Authors: Ishikawa Takuya , Kitaura Yasuyuki , Itami Yuya , Kadota Yoshihiro , Kato Rina , Inukai Ayami , Shimomura Yoshiharu
Published: June 2021

The catabolism of branched‐chain amino acids (BCAAs: leucine, isoleucine, and valine) is regulated by branched‐chain alpha‐ketoacid dehydrogenase complex (BCKDC), which is located at the second step of the catabolic pathway. The BCKDC is regulated by a covalent modification; the specific protein kinase BDK is responsible for inactivation of the complex by phosphorylation. In order to clarify the physiological significance of the BCAA catabolism in muscle tissues, we produced muscle‐specific BDK knockout (mBDK‐KO) mice. The mBDK‐KO mice showed the same growth rate at least until 10 weeks of age and no difference in weights of heart and skeletal muscles (soleus, gastrocnemius, and plantaris) obtained at 10 weeks of age, compared to those of normal mice. The BDK was not detected in skeletal and heart muscles but was present in other tissues of the mBDK‐KO mice. The E1 alpha component of the BCKDC was over‐expressed and was fully dephosphorylated in muscles of the mBDK‐KO mice, suggesting promotion of the BCAA catabolism. The mBDK‐KO mice did not show abnormal behavior observed in the mice deficient in BDK in whole body, suggesting no neurological abnormalities in the mice. The mBDK‐KO mice may be a useful animal model for the study of BCAA functions and metabolism in muscles.
Carbohydrate energy‐replacement attenuates protein turnover and promotes nitrogen retention during acute periods of increased metabolic demand (820.8)
Authors: McClung Holly , Lin Greg , Smith Tracey , Bukhari Asma , Lieberman Harris , Pasiakos Stefan
Published: June 2021

Acute periods of increased metabolic demand and subsequent energy deficit increase protein loss. To examine whether consuming an energy‐replacement based primarily on carbohydrate (85%) with negligible protein (2%) attenuates protein loss during highly metabolic energy flux, whole‐body protein turnover (15N‐glycine) was measured in 23 adults at baseline and after 2‐days of increased metabolic energy expenditure ([mean ± SD] 1800 ± 350 kcal·d‐1) once in energy balance (BAL) and on a separate occasion during energy deficit (DEF, 90%). Turnover was the same at baseline for both trials. After DEF, protein flux, synthesis, and breakdown rates were higher and net balance was lower than after BAL (time‐by‐energy, P < 0.05). Within trials, protein flux decreased for BAL but remained constant for DEF (time‐by‐energy, P < 0.05). Synthesis and breakdown decreased over time during BAL, whereas synthesis remained steady and breakdown increased 36% during DEF (time‐by‐energy, P < 0.05). Thus, net protein balance was maintained during BAL but was negative at the end of DEF (‐0.7 ± 0.4 g·kg‐1·d‐1; time‐by‐energy, P < 0.05). These data confirm the detrimental effects of energy deficit on protein balance and indicate that, by attenuating protein turnover, a carbohydrate‐based energy replacement can promote nitrogen retention during acute periods of increased metabolic demand.

Grant Funding Source: USAMRMC and DMRDP
Docosahexaenoic acid synthesis rates exceed brain uptake rates in the rat (821.13)
Authors: Bazinet Richard , Domenichiello Anthony , Chen Chuck , Trepanier Marc , Stavro Mark
Published: June 2021

Docosahexaenoic acid (DHA) is important for brain function, however, the exact amount needed for the brain is not agreed upon. While it is believed that the synthesis rate of DHA from α‐linolenic acid (ALA) is low, how this synthesis rate compares with the amount of DHA required to maintain brain DHA levels is unknown. The objective of this work was to assess whether DHA synthesis from ALA is sufficient for the brain. To test this, rats consumed a diet low in n‐3 PUFA, or a diet containing ALA or DHA for 15 weeks. Over the 15 weeks, whole body and brain DHA accretion were measured, while at the end of the study, whole body DHA synthesis rates, brain gene expression and DHA uptake rates were measured. Despite large differences in body DHA accretion there was no difference in brain DHA accretion between rats fed ALA and DHA. In rats fed ALA, DHA synthesis and accretion was 100 fold higher than brain DHA accretion of rats fed DHA. Also, ALA fed rats synthesized approximately 3 fold more DHA than the DHA uptake rate into the brain. This work indicates that DHA synthesis from ALA may be sufficient to supply the brain.

Grant Funding Source: Bunge Ltd
Does production of methane in the gut affect glucose tolerance? (822.10)
Authors: Rust Bret , Horn William , Cooper Danielle , Welch Lucas , Martin Roy , Pelkman Christine , Kim Eun Bae , Marco Maria , Keim Nancy
Published: June 2021

Intake of dietary fiber, which influences the composition of the gut microbiota, falls well below recommended levels in Americans. Gut microbiota show tremendous heterogeneity across populations and some profiles may have the ability to influence glucose tolerance. Methanogenic bacteria can be abundant in high fermenting individuals which may affect glucose homeostasis. To investigate this possibility, 22 individuals were provided all food for 2 wk at a dietary fiber level of 4.9 ±0.4 g/1000 Kcal/d. On day 5, 6 or 7 of the 2nd week, a test day protocol was conducted with low fiber breakfast and lunch meals. Breath hydrogen and methane were measured hourly for 9 h. Twelve blood draws were taken over the nine hours to assess postprandial plasma glucose and insulin to both meals. The composition of the intestinal microbiota was also determined by sequencing 16S rRNA gene amplicons from fecal samples. Individuals with high levels of methanogenic bacteria (MP) were compared to those with low levels of methanogenic bacteria (NM). Glucose AUC was significantly lower in MP than in NM in the 4‐h post‐breakfast period but insulin was not. These data suggest that increased methane production under conditions of a low fiber diet may improve glucose tolerance due to higher fermentation of available energy and not through increased secretion of insulin.

Grant Funding Source: Funded by Ingredion, Inc. and USDA intramural funds.
Multimechanistic combination enhances selenium’s antiproliferative effect in prostate cancer cells (826.2)
Authors: Li Xiuqi , Manwaring Eliza , Hopkins Tyler , Worden Joseph , Schille Colton , Eggett Dennis , Lawson John , Christensen Merrill
Published: June 2021

The anti‐cancer activity of selenium (Se) has been demonstrated by multiple in vitro and in vivo studies. However, none have sought to combine different forms of Se, with their varied mechanisms, to enhance Se’s chemopreventive efficacy. In this study, we hypothesized that systematically blending multiple forms of Se will produce a pleiotropic, multimechanistic combination that optimizes the in vitro growth inhibition of PC‐3 cells. In order to accomplish this, we first determined the IC50s of methylseleninic acid (MSA), sodium selenite, and Se nanoparticles (nano Se) via Alamar Blue proliferation assay. They were 5.0 μM, 14.0 μM, and 14.6 μM, respectively. We then utilized mixture designs and response surface methodology to direct our combination experiments: 10 combinations of MSA, sodium selenite, and nano Se were selected, each equaling 10 μM Se total. Experimental data of these 10 combinations were used to construct a polynomial model. This mathematical model predicted the optimum combination of the these Se compounds in inhibiting PC‐3 cell growth to be 4.1 μM of MSA, 5.1 μM of sodium selenite, and 0.8 μM of nano Se. These data provide proof of principle that combinations of different Se compounds exhibit greater chemopreventive efficacy than individual Se agents.

Grant Funding Source: Supported by NIH CA122235 to MJC and BYU Cancer Research Center Fellowship to XL.
Gestational effects of maternal folic acid supplementation on DNA methylation in newborn rat pups (827.11)
Authors: Ly Anna , Ishiguro Lisa , Kim Denise , Im David , Kim Sung‐Eun , Sohn Kyoung‐Jin , Kim Young‐In
Published: June 2021

Gestational effects of maternal folic acid supplementation on DNA methylation in newborn rat pups Anna Ly, Lisa Ishiguro, Denise Kim, David Im, Sung‐Eun Kim, Kyoung‐Jin Sohn, Young‐In Kim University of Toronto & St. Michael’s Hospital DNA methylation is reprogrammed in utero and maternal folic acid (FA) supplementation has been shown to modulate DNA methylation in the offspring. We determined which gestation period is most sensitive to the effect of maternal FA supplementation on DNA methylation. Pregnant rats were randomized to 5 FA interventions: a control diet throughout pregnancy, FA supplementation at 2.5x the control in 1st, 2nd, or 3rd week gestation only or throughout pregnancy. Pups were killed at birth and blood, brain, liver, kidney and colon processed. Pups from dams fed the control diet throughout pregnancy had significantly lower plasma folate levels compared to the FA supplemented groups (p<0.001). Pups from dams supplemented with FA during the 2nd or 3rd week gestation or throughout pregnancy had significantly higher brain folate levels compared to other groups (p<0.001). Pups from dams supplemented with FA throughout pregnancy had significantly higher liver folate levels compared to other groups (p=0.005). Brain global DNA methylation incrementally decreased from early to late gestational FA supplementation and was the lowest in pups from dams supplemented with FA throughout pregnancy (p=0.026). Liver global DNA methylation was lower in pups from dams supplemented with FA throughout pregnancy compared with other groups (p=0.07). No significant difference in folate levels and DNA methylation was observed in the kidney or colon among the 5 groups. Pups from dams supplemented with FA in late gestation or throughout pregnancy had lower Er‐, Gr and Ppar‐ gene expression in the liver compared to other groups (p<0.05). Our data suggest that maternal FA supplementation affects tissue folate levels, DNA methylation and gene expression in an organ and gestation‐specific manner.
Influence of folate on lipid accumulation in human primary subcutaneous adipocytes (827.8)
Authors: Hausman Dorothy , Alvis Courtney , Bailey Lynn , Park Hea Jin
Published: June 2021

Epidemiological studies report inverse associations between serum folate concentrations and body mass index, however the potential influence of changes in folate status on adipose cell development and metabolism has only been examined in animal models or a murine cell line. The objective of this study was to determine the influence of alterations in folate availability on differentiation of human primary preadipocytes. Human primary preadipocytes, derived from subcutaneous adipose tissue of normal weight female donors (ZenBio Inc), were exposed to 0.06, 6 or 60 uM folic acid (FA; Sigma), 5‐methyltetrahydrofolate (MTHF; Merck) or tetrahydrofolate (THF; Merck) during the differentiation period. The impact of treatment on lipid accumulation was determined using AdipoRedTM Adipogenesis assay reagent (Lonza). Results indicate that FA, MTHF and THF decreased lipid accumulation in mature adipocytes, after adjusting for cell number. Interestingly, MTHF and THF, reduced forms of folate, exhibited a greater inhibitory effect on lipid accumulation. These results indicate that biologically active reduced forms of folate may have a greater effect on adipocyte differentiation in human adipocytes. This study also supports preliminary findings in murine cell lines demonstrating an effect of changes in folate availability on adipose cell development.

Grant Funding Source: Funded in part by HATCH #GEO00707. MTHF and THF were a kind gift from Merck & Cie, Switzerland.
Cinnamon polyphenols increase oxygen‐glucose deprivation of decreased PGE2 production by upregulation of SIRT1 and alleviation of the anti‐inflammatory effects (830.11)
Authors: Qin Bolin , Anderson Richard
Published: June 2021

Prostaglandin E2 (PGE2) is an important mediator of many biological functions, including vasodilation, both anti‐ and proinflammatory actions, and modulation of sleep/wake cycles. It is well known that cinnamon polyphenols have anti‐oxidant properties and that oxidative stress damages endothelial cells. We investigated the mechanisms underlying protective effects of an aqueous extract of cinnamon, high in polyphenols, in 3 hr oxygen‐glucose deprivation (OGD) treated mouse brain microvascular endothelial (bEnd.3) cells. Immunoprecipitation/Western blotting and immunofluorescent analyses showed that OGD/reperfusion down regulated PGE2 secretion into the media and cellular PGE2 expression. Cinnamon polyphenols reversed these damages and also induced sirtuin (SIRT)1 expression. Further studies showed that the effects were reversed by a SIRT1 inhibitor IV. Cinnamon polyphenols also inhibited the expression of the anti‐apoptotic protein, Bcl‐xl, and induced the expression of the pro‐apoptotic protein, Bax, in OGD treated cells. The OGD‐induced inflammatory factors, TNF‐α and phospho‐Nf‐kB p65, were suppressed by cinnamon polyphenols. Taken together, these results suggest protective roles of cinnamon on PGE2 production and that cinnamon polyphenols may be neuroprotectants for ischemia‐related injury.

Grant Funding Source: This work was supported in part by a USDA Cooperative Research and Development Agreement (CRADA No.5
The metabolic challenge induced by hypercaloric diet leads to increased activity of the HPA axis and cardiac autonomic dysfunction in rats with low glial angiotensinogen (831.3)
Authors: de Lima Daniel , Nascimento kenia , Shaltout Hossam , Coimbra Candido , Diz Debra
Published: June 2021

We assessed the effect of metabolic challenge induced by hypercaloric diet on neuroendocrine parameters, renin‐angiotensin system components and heart rate variability (HRV) in transgenic rats with low glia‐derived angiotensinogen [TGR(ASrAogen)]. ASrAogen rats were fed with chow diet (AO, n=12) or hypercaloric diet (AO‐HD, n=14) from 4 to 13 wks of age. Sprague Dawley rats served as controls (SD, n=15; SD‐HD, n=13). AO‐HD displayed two fold increases in circulating angiotensin II and ACTH compared to AO (p<0.05). Both AO groups had higher levels of circulating corticosterone (200%, p<0.05). In addition, AO‐HD animals exhibit elevated resting mean arterial pressure (AO‐HD, 93 + 2 mmHg; AO, 74 + 3 mmHg; p<0.05), associated with impaired baroreflex sensitivity (42%, p<0.05). Furthermore, AO‐HD had lower vagal (Seq up, 48%; HF alpha, 41%; p<0.05) and sympathetic (Seq Down, 35%; LF alpha, 55%; p<0.05) indices for control of HR. To summarize, ASrAogen rats under metabolic challenge exhibit elevated arterial pressure and cardiac autonomic imbalance, accompanied by increased circulating angiotensin II and increased activity of the HPA axis. We concluded that the glial renin‐angiotensin system is important for regulation of neuroendocrine and cardiac function in response to metabolic challenge induced by hypercaloric diet. Support: CNPq; NIH HL51952

Grant Funding Source: CNPq; NIH HL51952
Correlation of lipoprotein epoxide content to microvascular function after short‐term walnut intake (831.5)
Authors: Holt Roberta , Jung Yim Sun , Shearer Gregory , Keen Carl , Djurica Dragana , Newman John , Shindel Alan , Hackman Robert
Published: June 2021

We examined the relationship of walnut intake and walnut derived fatty acids on outcome measures of vascular health. Hypercholesterolemic postmenopausal women (n=36) were randomized to 4 weeks of 5 or 40g of daily walnut intake. Outcomes were measured after an overnight fast and 4 h after walnut intake, prior to and after the 4 week intervention. Forty g of walnut intake increased polyunsaturated fat intake, including n‐3 and n‐6 fatty acids. After 4 weeks of 40g of walnut intake, microvascular function as assessed by reactive hyperemia index (RHI) was significantly increased, while the augmentation index at a heart rate of 75, a measure of pulse wave reflection and arterial stiffness, was significantly decreased. No significant changes in vascular function were observed with 5g of walnut intake. Four weeks of 40g of walnut intake significantly reduced LDL levels, while increasing the lipoprotein content of alpha‐linolenic acid (ALA) and linoleic acid (LA). A number of ALA and LA oxylipins were increased within lipoproteins, particularly HDL. The changes in 12(13)‐epoxyoctadecamonoenoic acid (ρ = 0.78) and 15(16)‐epoxyoctadecadienoic acid (ρ=0.70) epoxides of LA and ALA, respectively, within HDL were significantly correlated to improvements in RHI. In conclusion, short‐term walnut intake increased microvascular function, possibly through changes to specific LA and ALA oxylipins within lipoproteins.