bluuebunny/crossref_metadata_2025 huggingface dataset:
Composition and stability of phytochemicals in five varieties of black soybeans (Glycine max)
Authors: Correa Camila Renata , Li Lei , Aldini Giancarlo , Carini Marina , Chen C‐Y Oliver , Chun Hye‐Kyung , Cho Soo‐Muk , Park Ki‐Moon , Russell Robert , Blumberg Jeffrey , Yeum Kyung‐Jin
Published: June 2021

The phytonutrient composition of 5 varieties of black soybeans (Glycine max) and their stability at 22, 4 and −80°C over 14 mo were determined by HPLC systems with electrochemical and UV detection. Polyphenol content was confirmed by an orbitrap high resolution mass spectrometer. The different varieties of black soybeans contained 249–405, 84–137, and 6.76–14.98 μg/g dry wt, respectively of α‐tocopherol, γ‐tocopherol, and lutein. The major polyphenols in black soybeans were daidzein and genistein at 193–288 and 145–223 μg/g dry wt, respectively. No significant decrease was found in total phenols across the temperatures and time tested. In contrast, lutein, α‐tocopherol and γ‐ tocopherol degraded significantly within 1 mo of storage at 22°C (p<0.01) but remained stable through 6 mo at 4°C and 14 mo at −80°C. Thus, black soybeans are rich source of α‐ and γ‐tocopherol, total phenols, daidzein and genistein with their content of these compounds varying dependent upon their variety. Storage of black soybeans at low temperature is recommended to reduce the loss of fat‐soluble nutrients over an extended period of time. [Supported by USDA ARS #1950‐51000‐065‐08S, USA and BioGreen 21 #20070301034009 RDA, Korea]
Dietary supplementation with green tea EGCG at high dose promotes inflammatory response
Authors: Pae Munkyong , Ren Zhihong , Meydani Mohsen , Shang Fu , Meydani Simin Nikbin , Wu Dayong
Published: June 2021

Epigallocatechin‐3‐gallate (EGCG) has been shown to have anti‐inflammatory activity. The majority of supporting evidence is from cell‐based in vitro studies in which EGCG is added at levels often unachievable by oral intake. The in vivo studies utilizing disease model animals have shown inconsistent findings. To determine the anti‐inflammation efficacy of and the body's tolerance to dietary EGCG, we fed C57BL/6 mice a diet containing 0, 0.15, 0.3, or 1% (w/w) EGCG for 6 wk. EGCG supplementation dose‐dependently reduced weight gain. Contrary to the assumption that EGCG would reduce the inflammatory response, mice fed 0.15 and 0.3% EGCG diet showed no change, whereas those fed 1% EGCG diet had even higher levels of inflammatory response. Splenocytes and macrophages (MΦ) showed a higher capacity to produce proinflammatory cytokines TNF‐α, IL‐6, and IL‐1β and lipid inflammatory mediator prostaglandin E2, and splenocytes showed a lower capacity to produce anti‐inflammatory cytokine IL‐4. Furthermore, spleens from mice fed 1% EGCG diet had higher proportions of regulatory T cells, MΦ, natural killer (NK) cells, and NKT cells compared to those from mice fed other diets. These results suggest that high intake of EGCG may induce an inflammatory response, and this change may be associated with a disturbed homeostasis of immune cells involving the change in both function and number of specific populations. While the mechanisms and clinical significance for this effect of EGCG require further investigation, our results suggest that taking high doses of EGCG may cause an inflammatory effect rather than inhibit it. Supported by USDA #58‐1950‐7‐707.
Diet Quality of Americans 65 Years and Older
Authors: Hiza Hazel A.B. , Guenther Patricia M. , O'Connell Kellie , Davis Carole A
Published: June 2021

Dietary improvement can lead to reduced disease risk and improved health in older adults. This study examined the diet quality, measured by the Healthy Eating Index‐2005 (HEI‐2005), of older Americans 65 years and older using 24‐hour dietary recall data from the 2003–2004 National Health and Nutrition Examination Survey. The HEI‐2005 is a dietary assessment tool that measures quality in terms of how well diets conform to the 2005 Dietary Guidelines for Americans. Data were analyzed using SAS 10.1 and SUDAAN 10.0 (PROC ratio). In 2003–2004, for older Americans, the average HEI‐2005 scores met or exceeded the diet quality standards for whole fruit, total grains, and meat and beans. However, scores fell short of standards for total fruit (86%); total vegetables (82%); dark green and orange vegetables and legumes (34%); whole grains (32%); milk (56%); and oils (76%). Average intakes of saturated fat, sodium, and calories from solid fats, alcoholic beverages, and added sugars were higher than desired, resulting in scores of 62%, 34%, and 35%, respectively. For these components higher intakes result in lower scores. The diet quality of older Americans could be improved by choosing more foods/beverages that are lower in sodium and have fewer calories from solid fats, added sugars, and alcohol, and eating patterns that include more fat‐free milk, vegetables, fruit, whole grains, and oils. Source of support: USDA.
Binding of ΔNp63 to the promoter of p21, a p53‐dependent cell cycle arrest gene, modifies the response of salivary glands to therapeutic radiation
Authors: Mitchell Geoffrey C , Fillinger Jamia L , Avila Jennifer L , Victory Kerton R , Limesand Kirsten H
Published: June 2021

Radiotherapy targeting head and neck tumors often results in persistent loss of function in the salivary glands, leading to severe malnutrition other debilitating side effects in these patients. Radiation‐induced salivary gland damage results from attrition of salivary cells to p53‐dependent apoptosis. In p53 knockout mice as well as in mice expressing a constitutively active form of Akt (myr‐Akt1), this apoptosis is suppressed, and slaivary gland function is maintained. Injections of IGF1, a potent activator of Akt in salivary glands, also results in preservation of irradiated glands. Induction of cell cycle arrest may be important for this protection by allowing cells time for extensive DNA repair. We have observed increased accumulation of cells in G2/M and a reduction in proliferation at acute time‐points following irradiation in salivary glands of mice receiving pre‐treatment with IGF1. IGF1 also leads to a decreased stress response in irradiated glands as indicated by reduced stress‐induced phosphorylation of p53 (Ser18) and ΔNp63 (Ser160/162). The ΔN isoforms of p63 are thought to act as dominant negatives to p53‐induced gene expression. Given that several p53 target genes, including p21, are involved in cell cycle regulation, this suggests a mechanism by which IGF1’s affects on p63 could modulate arrest. Interestingly, pre‐treatment with IGF1 reduces binding of ΔN to the p21 promoter following irradiation, which may allow p53 to transactivate that gene and initiate arrest and repair, resulting in long‐term preservation of salivary gland function. Support: NIDCR K22 (DE16096), CBIO T32 (CA09213), NSF GK‐12 (0338247).
The modulation of intestinal endotoxin transport by different dietary fats
Authors: Mani Venkatesh , Gabler Nicholas K
Published: June 2021

Intestinal derived endotoxin and the subsequent endotoxemia are considered to be major predisposing factors for diseases such as atherosclerosis, sepsis, obesity and diabetes. Therefore, the aim of this study was to assess the effects of dietary fat on intestinal endotoxin transport. We hypothesized that saturated fat (SF) would augment, while n‐3 fatty acids (FA) would attenuate endotoxin transport across the small intestine of pigs. Eleven pigs (three‐four weeks of age) were euthanized and jejunum segments were mounted in modified Ussing chambers. Segments were treated with either no oil (CON), 12.5% (v/v) vegetable oil (VO), DHA (DO), cod liver oil (CLO), coconut oil (CO) or olive oil (OO) in 20 mM bile acid. Ex vivo endotoxin transport was measured by the addition of fluoroscein isothiocyanate labeled lipopolysaccharide (FITC‐LPS, 20 μg/mL) and serial sampling of the mucosal and serosal chambers. Fluorescence was measured in a plate reader and an apparent permeability coefficient (Papp) was calculated using the surface area and transport rate. Compared to the CON, DO and CLO reduced the endotoxin Papp by 50% (P<0.01). However, versus the CON, CO increased the Papp by 60% (P=0.008). OO and VO Papp were not different from the CON. Overall, these data indicate that fat high in n‐3 FA can attenuate intestinal endotoxin transport, while SF augments endotoxin transport. This work was supported by the USDA and NWRC.
Endocannabinoid (EC) system expression in osteoblast like cells is influenced by differentiation and eicosapentaenoic acid treatment
Authors: Hutchins Heather Lea , Li Yong , Hannon Kevin , Watkins Bruce A
Published: June 2021

The cannabinoid receptor, CB2 is expressed at increasing levels with osteoblast differentiation. Anandamide (AEA) is an endogenous agonist for cannabinoid receptors. Previously, we reported that arachidonic acid (AA), the substrate for n‐acyl phosphatidylethanolamine phospholipase D (NAPE‐PLD) for the synthesis of AEA, increased NAPE‐PLD expression compared to n‐3 PUFA treatment (EPA+DHA) in osteoblast‐like cells. The aim of this study was to determine if fatty acid treatments that alter enzyme substrate concentrations modify EC system expression and osteoblast markers. MC3T3‐E1 cells were grown for 6, 10, or 20 d in osteogenic medium; 72 h prior to collection cells were treated with 10 μM of AA, EPA, DHA, oleic acid (OA), or vehicle control. Increased NAPE‐PLD, FAAH, and CB2 mRNA expression and greater alkaline phosphatase (ALP) activity were observed with longer culture time. While CB1 expression was barely detectable and did not increase with time in culture, AA and EPA treatment increased CB1 compared to the other groups. NAPE‐PLD expression was lower in osteoblast‐like cells treated with EPA compared to all other groups. CB2 also decreased with EPA treatment. These results reiterate the impact of osteoblast maturation on NAPE‐PLD and CB2 expression. EPA might hinder EC synthesis and CB2 receptor expression during osteoblast differentiation. Supported by USDA.
Contribution of Inflammatory Markers to Changes in Bone in Postmenopausal Women: A 1‐Year Investigation
Authors: Gertz Erik R , Silverman Natalie , Wise Kristine S , Stewart Jeanne W , Hanson Kathy B , Alekel D Lee , Van Loan Marta D
Published: June 2021

Bone formation and resorption can be influenced by inflammation as reflected by inflammatory markers. The objectives of our study were to examine the relationships among inflammatory markers and bone mineral content (BMC) and density (BMD) and determine the contribution of inflammatory markers to 1‐year changes in BMC and BMD in healthy postmenopausal women. Data from the Soy Isoflavones to Reduce Bone Loss study (n=235) were examined. BMC and BMD from the lumbar spine & total proximal femur (hip) were measured by dual energy x‐ray absorptiometry and 4% tibia by peripheral quantitative computed tomography. Inflammatory markers (C‐reactive protein (CRP), interleukin (IL)‐1β, IL‐6, TNF‐α) were measured at baseline, 6 & 12 mos. Significant negative associations among inflammatory markers (Il‐6, TNFα) were observed at 6 & 12 mo. for % change in hip, spine, and whole body BMC and BMD. There were no significant associations with inflammatory markers for 4% tibia BMC or BMD. Multiple regression analysis was used to model the contribution of inflammatory markers to %change in BMC and BMD, accounting for ~ 2–7% of the variance. Inflammatory markers made a small but significant contribution to the observed changes in BMC and BMD in healthy postmenopausal women.

Research was supported by NIH‐NIAMS (RO1 AR046922), USDA/ARS, WHNRC, NWRC, ISU; CTSC, CCRC, UCD (1M01RR19975‐01), National Center for Medical Research (UL1 RR024146).
Leucine Supplementation of a Low Protein Meal Increases Skeletal Muscle and Visceral Tissue Protein Synthesis in Neonatal Pigs by Stimulating mTOR‐Dependent Translation Initiation
Authors: Torrazza Roberto Murgas , Suryawan Agus , Gazzaneo Maria C. , Orellana Renan A. , Nguyen Hanh V. , Davis Teresa A.
Published: June 2021

Leucine appears to be the most effective single amino acid to trigger the activation of translation initiation factors that regulate protein synthesis in muscle. The effects of Leu in visceral tissues are not well understood. We hypothesized that enteral Leu supplementation of a low protein diet (LP) in neonatal pigs will acutely increase skeletal muscle and visceral tissue protein synthesis to a rate similar to that achieved by feeding a high protein diet (HP). Fasted 5‐d‐old piglets (n=21) were gavage fed at 0 and 60 min either: 1) LP diet, 2) low protein diet supplemented with Leu (LP+L) to equal HP diet, or 3) HP diet. Diets were isocaloric and lactose was equal. Fractional protein synthesis rates and translational control mechanisms were examined 90 min after feeding. Protein synthesis increased in longissimus dorsi, gastrocnemius, masseter, heart, jejunum, kidney and pancreas, but not liver, of the LP+L group compared to LP group alone and did not differ from HP group. LP+L and HP increased translational efficiency and phosphorylation of S6K1 and 4E‐BP1 in all tissues except liver. Our results suggest that low protein diets supplemented with leucine stimulates protein synthesis in muscle and most visceral tissues to a rate similar to that achieved by high protein diets and this stimulation involves activation of mTOR downstream effectors. (Supported by Ajinomoto Amino Acid Research Program and USDA 58‐6250‐6‐001).

Grant Funding Source: Ajinomoto Amino Acid Research Program and USDA 58‐6250‐6‐001
Atmospheric hypoxia increases bone robusticity in the American alligator
Authors: Owerkowicz Tomasz , Andrade Fernando , Elsey Ruth , Hicks James
Published: June 2021

During vertebrate evolution, atmospheric oxygen (O2) level may have varied from as low as 12% to as high as 30%, but no studies to date have considered the effect of ambient O2 on skeletal plasticity. We incubated eggs and subsequently grew alligator hatchlings under chronic hypoxia (12% O2), normoxia (21% O2) and hyperoxia (30% O2). Animals received monthly injections of fluorescent dyes to determine bone deposition rates. After three months, animals were sacrificed and their femora either sectioned at mid‐diaphysis, or ashed. We found femora of hypoxic alligators to have significantly greater mass‐specific cross‐sectional area (+15%), second moment of area (+20%) and polar moment of inertia (+23%) than those of either normoxic or hyperoxic hatchlings. Mineral content was also significantly higher (+6%) in bones of hypoxic animals. This suggests that exposure to chronic hypoxia, but not hyperoxia, resulted in increased resistance to compressive, bending and torsional stresses on the skeleton. Furthermore, the relationship between body mass growth and periosteal deposition rate was different between treatments groups, with hypoxic animals accruing more bone per unit body mass. We suggest that prevalent atmospheric O2 level need be considered when reconstructing size and growth curves of extinct vertebrates. Funded by the NSF IOB 04445680 and IOS 0922756 to JWH.
Cardiac shunting does not constrain aerobic capacity of the American alligator
Authors: Owerkowicz Tomasz , Eme John , Gwalthney June , Blank Jason M. , Hicks James W.
Published: June 2021

Despite having a similar four‐chambered heart, crocodilians and birds differ widely in their aerobic capacity. We investigated whether this is due to differences in outflow tract design, which allows for cardiac shunting in crocodilians, but not in birds. We hypothesized that removal of cardiac shunt will improve aerobic capacity (VO2max) by forcing the entire right ventricular ejection volume to the lungs. We surgically occluded the left aorta in alligator hatchlings to render them incapable of shunting. After 17 months of growth and exercise training, we subjected alligators to a graded treadmill exercise test and measured their VO2max. We found no significant difference in VO2max between surgically‐altered (7.7 mlO2/kg/min) and sham‐operated (8.0 mlO2/kg/min) alligators. To further test whether cardiac shunting occurs during exercise, we measured blood flow in the left and right aortae. Shunting (net forward flow in the left aorta) occurred at rest in undisturbed animals, disappeared at the onset of forced exercise, and returned slowly during undisturbed recovery. This suggests cardiac shunting ability does not limit aerobic performance in crocodilians, because the shunt is turned off during exercise. We propose that non‐avian theropods may have continued to enjoy benefits of cardiac shunting, even as they evolved a more superior aerobic capacity. Funded by NSF IOB 04445680 to JWH.
Clinical measures of heat stroke recovery do not reflect tissue injury in a conscious rat model
Authors: Helwig Bryan G. , Leon Lisa R.
Published: June 2021

The time course of heat stroke (HS) thermoregulatory, circulatory and tissue injury responses during long term recovery are not well characterized. Using a conscious rat (Sprague‐Dawley, N=7/group) HS model, core temperature (Tc, ±0.1°C; radiotelemetry), kidney function (biliary urea nitrogen [BUN]; iStat), renal and hepatic injury (H&E staining) were examined through 10 days of recovery. We hypothesized that clinical biomarkers of HS (Tc and BUN) would not accurately reflect the extent of tissue injury during long‐term recovery. Compared to control animals, HS rats developed fever on day 1, 2, 3, 6, and 8 of recovery (37.6°C ± 0.0 vs. 38.1°C ± 0.0; p<0.01). Although the incidence of renal lesions increased through 10 days of recovery in HS rats, BUN levels were similar (12 mg/dL) in control and HS rats. Hepatic lesions were most pronounced on day 2, but resolved by day 10 of recovery. In conclusion, these data indicate that typical clinical biomarkers of HS recovery, such as Tc and BUN are not accurate indicators of recovery or tissue injury following HS. Research supported by USAMRMC. Author views not official US Army or DoD policy.
Weight Change among Mexican American Students Involved in an Intensive Intervention to Prevent and Treat Obesity
Authors: Palcic Jennette Lea , Johnston Craig A , Stansberry Sandra , El‐Mubasher Abeer , Ly Nguyenvu D , Reeves Rebecca , Foreyt John P.
Published: June 2021

The current study examined the effects of an intensive weight management intervention for Mexican American adolescents. A total of 228 adolescents were randomized to an environmental health promotion program (EHPP) or EHPP plus intensive intervention (EHPP+II). The EHPP consisted of a school‐wide intervention to promote healthy behaviors and included a physical education class. Children in the EHPP+II condition were exposed to the school‐wide intervention, but their PE class was replaced with an intensive multi‐component program comprised of nutrition and physical activity education, snacking intervention, and behavior modification. Overall, there was a significant reduction in zBMI among all students at 1 year (F= 40.75, p<.001) and there was a significant interaction (F= 3.31, p<.05). RANOVAs indicated no significant difference in change in zBMI between the EHPP+II and EHPP groups for healthy weight children (F=.967, ns). However, there was a significant interaction effect for overweight and obese children (F=3.45, p<.05). Both groups significantly decreased their zBMI at 6 months; however, at 12 months the students in the EHPP+II maintained their original losses, while students in the EHPP group increased their zBMI. This study suggests that a school‐wide health promotion program can reduce zBMI in overweight and obese children and may prevent increase in zBMI for normal weight children. Supported by the USDA.
Pregnancy and choline intake alter the metabolic use of orally consumed choline in women consuming deuterium labeled choline
Authors: Yan Jian , West Allyson , Perry Cydne , Jiang Xinyin , Yousef Leyla , Malysheva Olga , Devapatla Srisatish , Pressman Eva , Caudill Marie A.
Published: June 2021

A 12‐week choline intervention study was conducted to investigate the effects of pregnancy and choline intake on the fate of orally consumed choline. Healthy non‐pregnant (n = 21) and pregnant women (n=27, 27wk gestation) were randomized to controlled choline intakes of either 450 (choline AI) or 900 mg/day; 350 mg/day dietary choline and either 100 or 550 mg/d supplemental choline chloride. During the last six weeks of the study, ~20% of the total choline intake was provided as deuterium labeled choline (d9‐choline). The labeling of choline and its metabolites was examined at the end of the study (wk 12) in plasma and urine. Pregnancy affected the use of dietary choline with higher concentrations of plasma d9‐choline (P = 0.008), lower concentrations of plasma d6‐dimethylglycine (d6‐DMG) (P < 0.001), and higher urinary d9‐choline excretion (P = 0.001) in pregnant women. A higher choline intake (900 vs 450 mg/d) yielded higher plasma concentrations of d9‐choline (P = 0.003), d6‐DMG (P = 0.02), and d‐9 betaine (especially in pregnant women; P= 0.002 for choline x pregnancy) as well as greater urinary d6‐DMG excretion (P = 0.006). These data show that (i) pregnancy alters the use of orally consumed choline and (ii) a choline intake level exceeding the choline AI increases the availability of labile methyl groups for one‐carbon metabolic reactions especially in pregnant women.

Supported by grants from the USDA, ENC, and NCBA.
Analysis of a 25kD RNA Binding Protein from Leishmania tarentolae
Authors: Addy Nicole , Chen Zhiliang , Jagus Rosemary , Pitula Joseph S
Published: June 2021

Leishmania species are parasites belonging to the Trypanosomatidae family of protozoa. Currently, there is no effective vaccine against leishmaniasis. Thus, insight of underlying genetic regulation in these parasites will most likely aid in the development of effective chemotherapeutic interventions. Using the non‐pathogenic species L. tarentolae as a model organism, we cloned an RNA binding protein, p25. Characterization of p25 was undertaken to determine whether its specific binding targets were similar to those of the closely related trypanosomatid orthologues, Trypanosoma brucei p34 and p37. These proteins are known 5S rRNA binding proteins that play a central role in maintaining correct ribosomal concentrations in the cell. Using both recombinant protein and native extracts, we demonstrate that p25 binds to single stranded DNA with high affinity, consistent with other known RNA binding proteins. Direct gel shift analysis of p25 with labeled L. tarentolae 5S rRNA demonstrates that there is not a direct interaction between these molecules; however, the possibility remains that p25 interacts with 5S rRNA indirectly through a ribonucleoprotein processing particle. We maintain the hypothesis that p25 is likely to be involved in ribosomal biogenesis. Supported by NIH R25GM063775 and NSF 0614882
Snacking is Associated with Better Diet Quality and Reduced Risk of Overweight and Abdominal Obesity in Children
Authors: Nicklas Theresa A. , O'Neil Carol E. , Fulgoni Victor L
Published: June 2021

Snacking is very common among Americans; the impact of variety of snacking patterns on nutrient intakes and weight status is unclear. This study examined the associations of snacking patterns on nutrient intake and weight in children 2–18 years (n=14,220). Cluster analysis generated 12 distinct snacking patterns, explaining 57% of variance in total calories consumed. Only 8% of the children did not consume snacks. Cakes, cookies and pastries were the most common snacking pattern (16%) followed by miscellaneous snacks (e.g. whole milk, orange juice and meat/fish/poultry; 13%) and crackers and salty snacks (10%). Most snacking patterns resulted in higher total energy intake than no snack pattern. With higher energy consumption there were higher intakes of potassium; calcium; folate; fiber; riboflavin; zinc; vitamins A, C, B12 and K; and magnesium. However, most of the snacking patterns resulted in higher total intake of saturated fat, solid fat, added sugar and sodium (nutrients to limit). The majority of snacking patterns were associated with a reduced risk of overweight and abdominal obesity. Overall, several snacking patterns compared with non‐snackers had better diet quality and were less likely to be overweight or obese and less likely to have abdominal obesity. Education is needed to improve snacking patterns in terms of nutrients to limit in the diet. Funded by USDA and Kellogg's Corporate Citizenship Fund.
The effect of dietary fat and omega‐3 fatty acids on whole body lipid oxidation
Authors: Ainslie-Waldman Cheryl , Young Lindsay R. , Kurzer Mindy S. , Raatz Susan K. , Csallany A. Saari
Published: June 2021

Lipid peroxidation of polyunsaturated fatty acids yields several electrophilic, reactive carbonyl metabolites. We hypothesized that an increased intake of omega‐3 fatty acids (n‐3) would lead to increased lipid peroxidation metabolites compared to a diet low in n‐3. As part of a randomized crossover dietary intervention trial, urinary markers of whole body lipid peroxidation were analyzed. Study subjects (n=15) included healthy, postmenopausal women who were fed a low‐fat diet (20% of energy from fat, LF) or low‐fat diet supplemented with 3% of energy from n‐3 fatty acids (LFn3) for 8 weeks. Urine was collected during two consecutive 24‐hour collections in the last two days of each diet. Whole body lipid peroxidation within each subject was measured via complementary methods. Analysis via urinary TBARS was followed by a sensitive HPLC separation and quantification of 2,4‐ dinitrophenylhydrazine‐reactive lipophilic aldehydes, including 4‐ hydroxynonenal, 4‐hydroxyhexenal, and other nonpolar aldehydes. The HPLC technique demonstrated that LFn3 resulted in increased levels of lipid peroxidation products by two‐fold relative to LF. These results show that increased n‐3 intake leads to increased production of in vivo secondary lipid peroxidation products. The impact of these results requires further investigation. This work was supported by the US DOD, NCI, NCRR, and Salmon of the Americas, Inc.

Grant Funding Source: National Cancer Institute
Mathematical model gives insights into vitamin B6 and kynurenine metabolism
Authors: Rios-Avila Luisa , Nijhout H. Frederik , Reed Michael C. , Sitren Harry S. , Gregory Jesse F.
Published: June 2021

Vitamin B6 deficiency and oral contraceptive (OC) use have been associated with impaired tryptophan (trp) metabolism. To investigate the underlying mechanism, we developed a mathematical model of trp metabolism via the kynurenine pathway. The model includes kinetics from gut to liver, muscle and brain as well as trp concentrations in portal and peripheral circulation. Regulatory mechanisms and inhibition of relevant enzymes were included. Changes in urinary excretion of trp catabolites were observed as a function of vitamin B6 status using our model. Excretions of kynurenine, 3‐hydroxykynurenine and xanthurenic acid were increased in vitamin B6 deficiency as reported in the literature. Predicted effects of a trp load alone or concurrent with vitamin B6 deficiency also were consistent with experimental data. Dynamic variations of trp and trp metabolites were observed after simulation of trp input due to meals. Changes in trp metabolites concentrations were seen after induction of tryptophan 2,3‐dioxygenase alone or in combination with impaired B6 status as reported in OC users. Our findings support the use of this mathematical model as a tool to test trp metabolism in different conditions to complement, clarify and provide new insights into the associations of altered trp metabolism with OC use and vitamin B6 deficiency. Supported by NIH grant DK072398, and NSF grants DMS‐061670 (MR, HFN), EF‐1038593 (HFN. MR).
Characterization of circulating phosphatidylcholine fatty acids in non‐pregnant women with supplemental DHA and variable choline intake
Authors: West Allyson A , Jiang Xinyin , Perry Cydne , Yan Jian , Caudill Marie A
Published: June 2021

Phosphatidylcholine (PtdCho) synthesized via the phosphatidylethanolamine N‐methyltransferase (PEMT) pathway is enriched with polyunsaturated fatty acids (FAs) including docosahexaenoic acid (DHA). Methyl groups from the choline derivative betaine may be used by PEMT thus choline intake may influence the FA makeup of circulating PtdCho by modulating activity of biosynthetic pathways. DHA intake is associated with concentrations in blood, but it is unknown how varied choline intake may interact with supplemental DHA to impact PtdCho FAs. This study characterized plasma and RBC PtdCho FA species in non‐pregnant women consuming a DHA supplement and two levels of choline. The proportion of plasma and RBC PtdCho DHA increased from 3.4% and 2.6% to 4.9% and 3.4%, respectively. The ratio of other PtdCho FAs decreased concurrently and will be discussed. By study end, alterations of PtdCho FAs were independent of choline level; however, from baseline to week 6 women consuming more choline experienced a spike of plasma PtdCho DHA. Preliminary results indicate a higher choline intake combined with supplementary DHA may increase PEMT activity in the short term. Future studies elucidating the impact of choline intake and supplemental DHA on PtdCho synthesis via the PEMT pathway will include deuterium labeled PtdCho data. Funded in part by USDA, ENC and the Beef Checkoff; AAW is supported by an ENC Dissertation Fellowship.
Rapid screening for zinc deficiency using portable x‐ray fluorescence in fingernails
Authors: Fu Kristin A , Kehayias Joseph J
Published: June 2021

Elemental composition of fingernails is a useful indicator of micronutrient status and may reflect an individual's intake over time. Our objective was to determine if portable X‐ray fluorescence (XRF) is a viable method to assess zinc content in fingernails in the field. Human fingernail samples ranging from 3 to 85 mg were collected with zinc‐free steel clippers, cleaned and analyzed by non‐destructive XRF without any further preparation. Some specimens were also analyzed by indirect current plasma emission spectrometry to provide absolute calibration. Reproducibility was found to be better than 2% for samples >30 mg. Zinc was detected in human fingernails and animal hooves. XRF zinc values were compared against chemical and natural phantoms. Material from hooves served as high (swine, 135μg/g) and low (bovine, 52μg/g) standards. We concluded that portable, non‐destructive XRF is a feasible method to determine zinc content in fingernails and an alternative to atomic absorption/emission spectrometry allowing for simple sample preparation and fast, automated analysis. Applications include field screening for zinc deficiency at the population level or to measure the efficacy of an intervention. Work supported by the USDA, agreement No. 58‐1950‐7‐707.
Dietary modulation of liver lipogenic gene expression in obese female Zucker rats
Authors: Al-Dwairi Ahmed , Simmen Frank , Fuchs George , Korourian Soheila , Hakkak Reza
Published: June 2021

Obesity is a risk factor for breast cancer. Liver is a major lipogenic organ whose metabolism contributes to body fat mass. We reported earlier that obese female Zucker rats fed soy protein‐based diet gained more weight and developed more mammary tumors than did casein (CAS)‐fed rats after DMBA treatment. We investigated the state of hepatic lipogenic gene expression to test if tumor incidence/pathology was correlated with induced hepatic lipogenesis. Lean and obese female Zucker rats were fed soy protein or casein‐containing diets beginning at five weeks of age and were euthanized 147 days after DMBA treatment. Obese rats had increased liver weights compared to lean rats; the soy protein diet reduced liver weights in both the lean and obese groups. Liver transcripts encoding FASN, ME1 and SREBP‐1c were significantly induced (20–40%; P <0.05) in the obese soy‐fed rats when compared to obese CAS‐fed or the lean rats. The 6PGD gene was down regulated in the soy‐fed lean compared to CAS‐fed lean groups, but was induced with soy consumption in the obese group. Western blots of ME1, a major obesity associated gene, revealed a ~3‐fold induction in the obese soy‐fed group and which paralleled tumor incidence. Data highlight the importance of hepatic lipogenesis in obesity and tumor development, and suggest that intervention within this pathway may provide a way to reduce tumor incidence. (Supported by ABI, USDA and NIH)
Effects of whole eggs combined with other cooked breakfast foods on postprandial metabolism in older, overweight adults
Authors: McKay Diane L. , Chen C-Y. Oliver , Rasmussen Helen , Blumberg Jeffrey B.
Published: June 2021

Adverse postprandial metabolic changes may contribute to the pathogenesis of chronic diseases. Eggs provide highly digestible protein, unsaturated fatty acids, carotenoids and other antioxidants that may act synergistically to antagonize these effects when combined with unhealthy breakfast foods. To determine the postprandial metabolic effects of a typical cooked American breakfast high in glycemic load either with no eggs (N), 2 eggs (E), or a nutrient‐poor protein + cholesterol egg substitute (ES), we conducted a 3‐way crossover pilot study in 7 subjects (age 50–70 y, BMI 28–34) with a 2‐d low antioxidant diet run‐in and 1 wk washout for each intervention. Blood and urine were collected before and for 4 h after each isocaloric breakfast. Compared to baseline, GLP‐1 increased 135, 78 and 61% at 2 h with E, ES, and N, respectively (E vs. ES, P<0.05), while ghrelin decreased 6% for N (P>0.05 vs. E and ES, 16% for each). Plasma total thiols also increased at 2 h by 7.4, 5.5 and 0.5% with E, N, and ES, respectively (E vs. ES, P<0.05). At 2 and 4 h the increase in urinary isoprostanes was similar with each breakfast. Glucoregulation was better controlled with E compared to ES and N (HOMA‐IR: E<ES<N; AUCins/glu: E<ES0.05). These data suggest the positive postprandial effects of eggs on satiety and endogenous antioxidant defenses may be attributed to the whole food. (Supported by USDA and the Egg Nutrition Center)
RGD‐peptide lunasin inhibits PI3‐kinase/Akt‐mediated NF‐κB activation in human and murine macrophages through interaction with αvβ3 integrins
Authors: Cam Anthony , de Mejia Elvira Gonzalez
Published: June 2021

Regulation of aberrant macrophage activity under inflammatory conditions is critical in the prevention of cardiovascular disease (CVD). Integrin receptor αvβ3 is highly expressed on macrophages in atherosclerotic lesions and binds Arg‐Gly‐Asp (RGD) ligands. Lunasin is a food derived peptide that contains a RGD cell adhesion motif. The objective was to comparatively determine the effect of lunasin on pro‐inflammatory markers from LPS‐induced RAW 264.7 murine and THP‐1 human macrophages and its role on αvβ3 integrins, important mediators of PI3K/Akt activation of NF‐κB. Lunasin significantly inhibited NO (IC50 = 41 μM; <10 μM), iNOS (IC35 = 36 μM; IC50 <10 μM), COX‐2 (IC50 = 31 μM; <10 μM) and p‐Akt (IC35 = 29 μM; IC50 = 11 μM) in murine and human macrophages, respectively. In RAW 264.7, lunasin significantly inhibited PGE2 (IC50 = 48 μM) and reduced nuclear NF‐κB subunits p65 and p50 by 26% and 43% at 50 μM, respectively. Lunasin significantly reduced p‐p65 (IC50 <10 μM) and αvβ3 integrin expression (IC50 = 10 μM) in THP‐1. Vitronectin, a ligand of αvβ3 integrin, internalizes and increases expression of pro‐inflammatory markers, whereas a PI3K inhibitor attenuated these effects. After treatment of both cell lines with lunasin, co‐immunoprecipitation confirmed direct interaction between lunasin and αvβ3 integrin. Lunasin reduced integrin‐mediated inflammatory responses and has potential in the prevention of CVD.

Grant Funding Source : USDA
Iron status and variations in electroencephalography (EEG) during five cognitive tasks in Indian adolescents
Authors: Hammons Julie , Wenger Michael , Scott Samuel , Murray-Kolb Laura , Ghugre Padmini , Udipi Shobha , Boy Erick , Haas Jere
Published: June 2021

Iron deficiency (ID) is the most common micronutrient deficiency globally, and is associated with cognitive and perceptual deficits. Previous work has been limited in the combination of specific measures of cognition with simultaneous measurement of brain state in the context of human ID. The subjects were part of a randomized, double‐blind trial testing the efficacy of iron‐biofortified pearl millet as a mechanism to improve iron status. A cross‐sectional analysis of baseline data is presented here. Subjects (n= 75; 12–16 y) were from two residential schools in Maharashtra, India. Iron status was assessed by hemoglobin, serum ferritin, and serum transferrin receptor. Brain functioning was assessed via electroencephalography (EEG) while subjects completed 5 measures of perceptual and cognitive performance: simple reaction time, low‐ and high‐level attentional control, semantic memory, and recognition memory. These measures were chosen to analyze perceptual and cognitive components of schoolwork and for comparability to previous work in adults. We examined the relation of iron status to EEG for each task. Sub‐analyses by age and gender were also considered. Supported by Harvest Plus and NSF.
Initial Adipose Tissue Responses to High Milk‐fat Diet
Authors: Camell Christina , Smith Wayne
Published: June 2021

Obesigenic diets cause inflammation in adipose tissue (AT) including infiltration of lymphocytes and macrophages. Changes in macrophage polarization are observed, which apparently reflect the tissue microenvironment including available cytokines and fatty‐acids. The project goal is to analyze very early responses to a diet high in milk‐fat (MF) that is rich in palmitic (16:0) and oleic (18:1) acid. We have examined AT molecular and cellular changes after 3 days of MF feeding, and utilized genetically‐deficient mice to test if TLR4 is necessary for diet‐induced effects. The earliest responses in wildtype (WT) mice were limited to the mesenteric AT (MAT) and peritoneal leukocytes. There were increases in gene expression in MAT including monocyte chemotactic factor (MCP)‐1, chemokine (C‐X‐C motif) ligand (CXCL)‐1, secreted phosphoprotein (SPP)‐1 and the macrophage marker: F4/80. Additionally, we identified a decrease in the number of low and high expressing F4/80 positive peritoneal macrophages. TLR4−/− mice failed to exhibit diet‐induced increases in MCP‐1 in the MAT, and they failed to exhibit the decrease in peritoneal macrophages seen in WT mice. They also exhibited hyper‐expression of anti‐inflammatory macrophage markers in the MAT, suggesting a biased response to the MF diet. To test for fatty‐acid specific action in macrophages an in‐vitro model of fatty‐acid treatment was utilized. RAW 264.7 cells upregulated pro‐inflammatory macrophage markers (eg. TNF‐α) upon treatment with palmitic acid while oleic acid treatment upregulated anti‐inflammatory markers (eg. Arginase‐1). Treatment with a mixture of palmitic and oleic acid inhibited genes upregulated by treatment with a single fatty acid. These results suggest the early changes in response to a MF diet are localized in the mesentery and involve peritoneal cells, ATMs and their response to fatty‐acids.

Grant Funding Source : F31DK084841; USDA 6250‐51000‐055
Magnesium Oxide Nanoparticles Induce Cytotoxic and Proinflammatory Effects in Schwann cells and DRG Neurons
Authors: Idikuda Vinaykumar , Jaiswal Ashvin , wong Yin Yin wendy , leung solomon , Daniels christopher , lai james
Published: June 2021

Applications of metal oxide nanoparticles in diverse fields have been accelerating in the last decade. For example, magnesium oxide (MgO) nanoparticles have been employed in ceramics, and as adhesive and additive in chemical raw materials, leading to their increased human exposure, thereby posing potential health hazards. We have therefore investigated the hypothesis that MgO nanoparticles exert differential cytotoxic effects on neural cells in the peripheral nervous system employing cell models consisting of Schwann cells and dorsal root ganglion neurons (DRG) in vitro. Results of our ongoing studies revealed MgO nanoparticles exerted differential effects on lowering the survival of Schwann cells and DRG neurons. Along with their morphological changes, we also found the nanoparticles induced differential dose‐related increases in expression of markers of inflammatory responses such as Interleukin‐6 (IL‐6) and Peroxisome Proliferated Activated Receptor PPAR‐? in the two neural cell types. Thus, our results suggest that MgO nanoparticles can induce differential cytotoxic effects and elicit inflammatory responses in Schwann cells and DRG neurons and as such may have implications in health hazard of human exposure to the nanoparticles.

Supported by: USAMRMC project grant (contract#W81xWH‐07‐2‐0078) and NIH grant #P20 RR016454 from Idaho INBRE program of the National Center for Research Resources.
Overexpression of mitochondrial GSH transporters in renal proximal tubular cells from control and diabetic rats
Authors: Lash Lawrence Harold , Benipal Bavneet , Putt David Alan
Published: June 2021

We hypothesize that overexpression of mitochondrial glutathione (mtGSH) transporters in renal proximal tubular (PT) cells from diabetic rats will reverse the oxidative stress and diminish susceptibility to chemically induced cytotoxicity by producing sustained increases in mtGSH content. cDNAs encoding either the dicarboxylate (DIC; Slc25a10) or 2‐oxoglutarate (OGC; Slc25a11) carrier were expressed in renal PT cells from either control or streptozotocin‐induced diabetic Sprague‐Dawley rats using Lipofectamine. Transfection with either DIC or OGC cDNA produced 7,800‐ or 5,000‐fold increases in DIC or OGC mRNA, respectively, in cells from control rats and 600‐ and 1900‐fold increases in DIC or OGC mRNA, respectively, in cells from diabetic rats. Carrier overexpression provided significant protection, as judged by LDH release and MTT fluorescence, from injury due to the mitochondrial toxicant antimycin A. These results suggest that mtGSH carrier overexpression may be a viable approach to improving redox status of renal cells in chronic disease states.

(Supported by DOD Grant PR64340)
Post‐Exposure Therapy Administered 30 Minutes After an LD50 Dose of Soman in Guinea Pigs without Carbamate Pretreatment
Authors: DeBus Sandra J. , Spriggs David , Krempel Ryan , Chang Fat-Chun T.
Published: June 2021

Exposure to soman (GD), a chemical warfare agent (CWA), causes cholinesterase inhibition, seizure, convulsions, cardiorespiratory dysfunction, and death. Current US military management of CWA poisoning involves carbamate pretreatment and post‐exposure oxime/cholinolytic therapy. However, CWAs pose a potential threat to civilian populations without access to pretreatment programs. We used unanesthetized guinea pigs, instrumented to record CNS and cardiorespiratory activities, to determine a post‐exposure therapeutic window within which effects of GD poisoning could be mitigated without carbamate pretreatment. An LD99 dose of GD was followed by a therapy delay of up to 30 min. The therapy mix included im scopolamine (0.25 mg/kg), methylatropine (2 mg/kg), the oxime MMB4 (26.1 mg/kg), physostigmine (0.075 mg/kg), and ip Phenobarbital (25 mg/kg). After GD, all animals showed periods of hypersecretion, dystonia/tremor, seizure, bradycardia, and bradypnea. After therapy, cardiorespiratory parameters returned to control levels and seizures ceased within 10 min with no recurrence. 88% of animals survived >24 hrs. Of those, one was asymptomatic at 24 hrs, and the rest showed only mild dystonia. In conclusion, our therapy mix, even following 30 min of poisoning without pretreatment, effectively eliminated the most life‐threatening signs of GD intoxication. Funded by Defense Threat Reduction Agency.
Effects of age and lifetime behavioral patterns on locomotion in Drosophila
Authors: Roberts Stephen , Lane Steven , Mancinelli Georgina , Martinez Emily , Sandoe Laura , Kopke Danielle , Elekonich Michelle
Published: June 2021

The effects of behavior on physiology and senescence may be profound in variably active species such as flying insects, which attain among the highest metabolic rates ever measured. Previous studies have shown that limiting flight behavior in insects extends lifespan and slows the age‐related loss of antioxidant capacity and accumulation of oxidative damage in flight muscles. In this study, we manipulated the lifetime flight behavior of Drosophila melanogaster. Specifically, 5‐day old adult flies were separated into three life‐long treatments: (A) those not allowed to fly (no flight), (B) those allowed – but not induced – to fly (voluntary flight), and (C) those mechanically stimulated to perform over 100 flight bouts per day (induced flight). Consistent with the oxidative stress model of aging, Drosophila prevented from flying lived longer than those allowed or stimulated to fly. Flight capacity, measured as the ability to fly in a hypodense gas mixture (21% O2, 39.5% N2, 39.5% He; 0.81 g l−1), decreased with age in all treatment groups, but decreased earliest in flies from the no‐flight treatment. Extended longevity in flies from the no‐flight treatment suggests that oxidative damage accrued more slowly in this treatment group. However, early loss of flight capacity in the no‐flight group indicates that disuse effects were the primary determinant of the age‐dependent muscle impairment in this group. Additional experiments are ongoing to compare age‐dependent oxidative damage, flight muscle ultra‐structure, flight kinematics, immune function, reproduction and metabolic capacity among the treatment groups. Supported by NSF IOS‐0725030.
The quantitative genetics of a complex trait under continuous directional selection
Authors: Careau Vincent , Wolak Matthew , Carter Patrick A , GARLAND THEODORE
Published: June 2021

We analyzed data from a long‐term artificial selection experiment that includes 4 lines of mice bred for high voluntary wheel running (HR) and 4 non‐selected control (C) lines. The HR lines reached a selection limit at generation ~16, running ~3‐fold more revolutions/day than C lines. In addition, wheel running varied across generations in an apparently cyclical fashion in both HR and C. We used the first 25 generations to estimate quantitative genetic parameters before, during, and after the selection limit was reached. We used ASReml‐R to apply the “animal model”, a linear mixed‐model that uses all the information on the coefficients of co‐ancestry among individuals in a pedigree. Our preliminary results indicate additive genetic variance (VA) was not eliminated in HR lines after the limit was reached. However, the selection regime led to a negative covariance between VA and maternal genetic variance (VM), which could maintain VA in the selected trait and potentially explain the presence of a cycle. We also found that the genetic correlation between mean running speed and duration of wheel running tended to be lower in females than in males, which may explain why the response to selection was achieved differently in females (mainly speed) and males (both speed and duration). Supported by NSF grants IOS‐1121273 to TG and EF0328594 to PAC, and a NSERC postdoctoral fellowship to VC.