bluuebunny/crossref_metadata_2025 huggingface dataset:
Validation of Sun Exposure and Vitamin D Intake Recall Questionnaires for Estimating Vitamin D Status
Authors: Hall Laura Marie , Stephensen Charles B , Aronov Pavel A , Hammock Bruce D , Kimlin Michael G , Slusser James R
Published: June 2021

Individual sun exposure and dietary intake both contribute to vitamin D status but simple, validated recall methods to assess these exposures are lacking. The objectives of our study were to validate both a sun exposure recall questionnaire, relative to a validated daily exposure log, and a vitamin D‐specific food frequency questionnaire (FFQ), relative to food records. We then assessed the ability of these methods, in conjunction with skin reflectance, to predict serum 25‐hydroxy vitamin D (25OHD) measured by LC‐MS. The study involved 72 healthy young adults each followed for 7‐8 wks during each season in Davis, CA. Spearman's correlation coefficients and validity coefficients were used to validate the methods. Multiple linear regression was used to predict serum 25OHD using sun exposure, skin reflectance and diet. The recall methods were highly correlated to the reference methods (sun: r = 0.83‐0.95, diet: r = 0.69‐0.77, p < 0.0001) and moderately correlated to serum 25OHD (sun: r = 0.40‐0.51, diet: r = 0.36‐0.37, p < 0.005). The recall measurements were all significant predictors of serum 25OHD (p < 0.05) and were not significantly different than the reference measurements in the regression model. These results show that the contribution of sun and diet to vitamin D status can be assessed using simple recall methods which may be used by nutritionists to predict individual needs for vitamin D intake.

Grant Funding Source

NIH Grant P60 MD00222 and USDA Project 5306‐51530‐006‐00D
Atmospheric hypoxia constrains skeletal growth and alters bone microstructure in hatchlings of the American alligator (Alligator mississippiensis)
Authors: Owerkowicz Tomasz , Andrade Fernando , Elsey Ruth M. , Hicks James W.
Published: June 2021

Skeletal growth reflects whole‐body growth of the animal, and features of bone microstructure (vascular density, collagen arrangement) can be used to infer an animal's growth strategy. This relationship, however, may be altered by environmental change. Vertebrates are known to have endured large‐scale fluctuations in atmospheric %O2 during their evolutionary history, but no study to date has tested the effect of atmospheric %O2 on skeletal growth. We reared hatchlings of the American alligator under chronic hypoxia (12%O2), normoxia (21%O2) and hyperoxia (30%O2). Animals were fed ad libitum every other day, and received injections of fluorescent dyes to determine bone deposition rates. The femoral midshaft shows no significant difference in bone formation rates (BFR) between normoxic and hyperoxic alligators, and an identical correlation with body mass growth rate (GRMb). Hypoxic animals, however, show a significantly higher ratio of BFR:GRMb, which results in a relatively robust skeleton. This is surprising, since hypoxia is thought to impair collagen and mineral deposition in new bone. We argue that any inference of growth rates of extinct vertebrates based on their bone microstructure (e.g., on the amount of bone deposited between seasonal growth rings) should take into account the prevalent atmospheric %O2 contemporary for the species. Funded by NSF IOB 04445680 to JWH. TO supported by NIH 2T32AR047752.
Alterations in splenic HSP70 and IL‐6 associated protein expression following heat stroke in mice
Authors: Helwig Bryan Glen , Leon Lisa R
Published: June 2021

Plasma protein expression of HSP70 and IL‐6 are elevated following heat stroke (HS) in mice. The spleen is the largest secondary lymphoid organ in the body and we have shown this organ to be damaged following HS. Despite this, splenic expression of HSP70, IL‐6, and the IL‐6 receptor (IL‐6R) are not well understood. This study tested the hypothesis that HS would be accompanied by increased protein expression of IL‐6, IL‐6R, and HSP70 at HS collapse (Tc,Max=42.7°C), hypothermia depth (HD), and fever (24h) in splenic tissue. Spleens were harvested from control (n=4 per group) and HS mice at HS collapse (n=4), HD (n=4), and fever (n=4), protein isolated, and immunoblotting performed. Splenic HSP70 protein expression was only modestly increased at HS collapse but significantly elevated at HD through 24h of recovery. Splenic IL‐6 protein was not detected at any time point whereas IL‐6R was expressed, but not altered, at any point during or after HS. These data demonstrate that IL‐6 and IL‐6R protein expression are not significantly affected by HS or contribute to the development of splenic tissue damage. In contrast, splenic HSP70 protein expression is dynamic. Although HSP70 expression is significantly enhanced by HD and 24h the molecular chaperone is unable to protect the spleen against HS‐related tissue damage. Research supported by USAMRMC.

Author views not official US Army or DoD policy
Selected trace metals inhibit transepithelial non‐heme iron transport by inhibiting the basolateral iron exit in human intestinal cells.
Authors: Kim Eun‐Young , Ham Soo‐Kyung , Hao Sijie , Han Okhee
Published: June 2021

The relationship between iron and other trace metals has been recognized for many years. Intestinal absorption of other trace metals (e.g., Cd, Co, Zn and Mn) was significantly increased in iron deficient mice. On the other hand, other trace metals inhibit intestinal iron absorption. However, the mechanism by which these minerals interact in the absorption process is unknown. To investigate the interactions between iron and other trace metals in the process of intestinal absorption, we examined effects of selected trace minerals on the basolateral iron transport and apical iron uptake in fully differentiated Caco‐2 cells grown on microporouse membrane inserts. The addition of equimolar concentration of the selected trace metals (Cd, Co, Cu, Mn and Zn) significantly (P<0.05) inhibited the basolateral 55Fe transport. To examine if these metals decrease the basolateral iron transport by inhibiting the apical iron uptake, we next assessed effects of these minerals on the apical iron uptake. While the addition of Cu significantly (P<0.05) inhibited the apical 55Fe uptake, Cd, Mn and Zn did not influence the apical 55Fe import. Addition of equimolar concentration of Co significantly (P<0.05) increased the apical 55Fe uptake. These results suggest that some selected trace metals inhibit non‐heme iron absorption by decreasing the basolateral iron export rather than the apical iron uptake in intestinal cells. (Supported by NIH R21DK069946 and USDA: 2005‐35200‐16543.)
Role of Maternal/Fetal Iron Status on Placental Transferrin Receptor Expression
Authors: Young Melissa Fox , Foehr Marisa , McNanley Thomas , Cooper Elizabeth , Pressman Eva , Guillet Ronnie , Orlando Mark , McIntyre Allison , Lafond Julie , O'Brien Kimberly O
Published: June 2021

The placenta plays an important role in mediating iron transfer to the fetus. However, the responsiveness of the placenta to maternal and fetal demands remains unclear. Placental transferrin receptor (TfR) is a key placental iron transport protein on the apical membrane. Placentas were collected from twelve pregnant adolescents (38‐40 wks gestation) and protein expression of TfR was examined. Iron status indicators were assessed from maternal blood obtained during the second trimester and at delivery. Cord blood samples were also collected at birth. There was a significant inverse relationship between cord serum ferritin (SF) and placental TfR expression (P=0.042). A non‐significant inverse trend was detected between maternal SF in the second trimester and placental TfR expression (P=0.075). Relationships between maternal SF in the second trimester and cord SF approached significance (P=0.055). However, maternal SF at birth was not significantly related to placental TfR expression or cord SF. Hemoglobin and TfR in both the infant and mother were not significantly related to placental TfR expression. In conclusion both maternal and fetal iron status appear to impact placental expression of TfR. Studies are on‐going to assess the impact of maternal iron status on neonatal iron stores at birth in the planned cohort of 300 pregnant teens.

Grant Funding Source

Supported by USDA Grant no. 2008‐0857.
A diet high in meat protein and potential renal acid load increases absorption and urinary excretion of calcium, as well as serum IGF‐I in postmenopausal women
Authors: Cao Jay J , Johnson LuAnn K , Hunt Janet R
Published: June 2021

The objective was to determine the effect of increasing protein and potential renal acid load (PRAL) on Ca retention and markers of bone metabolism.

In a randomized crossover design, twenty postmenopausal women consumed two diets: one low protein, low PRAL (LPLP) and one high protein (mostly meat), high PRAL (HPHP) for 7 wk each, separated by a one‐week break. After 3 wk, the entire 2‐d menu of each diet was radio‐labeled with 47Ca and retention was measured by whole body scintillation counting for 4 weeks. Biomarkers of bone metabolism in blood and urine were measured.

Results

Compared with the LPLP diet, the HPHP diet increased urinary acidity (pH: 7.1 vs. 5.9, ±0.25*, p < 0.01), urinary Ca excretion (156 vs. 203, ±63 mg/d, p < 0.01), and blood IGF‐I levels** [137 (103 ‐ 184) vs. 174 (130 ‐ 232) ng/ml, p < 0.01) consistently from week 1 through 7. The fractional Ca absorption was lower in subjects with LPLP than with HPHP diet (25.0 vs. 30.4, ±5.4%, respectively, p < 0.02). The HPHP diet tended to increase the absolute amount of Ca absorbed compared with LPLP diet (227 vs. 258, ±47 mg/d, p < 0.08).

Conclusions

In postmenopausal women, a diet high in both meat protein and PRAL increased serum IGF‐I and Ca absorption, which compensated the increase in urinary excretion. A high meat diet does not appear detrimental for bone health.

*Pooled SD; **Geometric means (range).

Supported by USDA‐ARS and the National Cattlemen's Beef Association.
Cinnamon extract exhibits insulin‐like and independent effects on gene expression in adipocytes
Authors: Cao Heping , Anderson Richard A.
Published: June 2021

Cinnamon is beneficial to people with insulin resistance due in part to the insulin‐like activity of the cinnamon extract (CE). Molecular effects of CE are limited. This study tested the hypothesis that CE has insulin‐like and insulin‐independent effects at the molecular level. Quantitative real‐time PCR was used to evaluate CE effects on the expression of 43 genes coding for the glucose transporter (GLUT) family, insulin‐signaling components, adipokines, pro‐inflammatory cytokines and anti‐inflammatory tristetraprolin (TTP) family in mouse 3T3‐L1 adipocytes. CE (100 mg/L) increased GLUT1 mRNA levels by 2‐7 fold after 2‐16 h treatment. CE increased TTP mRNA levels by up to 10‐fold. Vascular endothelial growth factor (VEGF) mRNA levels were decreased 40‐50% by CE from 0.5‐4 h. CE decreased the expression of most genes encoding insulin signaling pathway proteins. Insulin‐like effects of CE included rapid induction of TTP mRNA, and reduction of mRNA for VEGF and components of the insulin‐signaling pathway. Insulin‐independent effects of CE included sustained increases in GLUT1 and TTP expression. These results demonstrated that CE regulated multiple gene expression in mouse adipocytes and suggested that the health benefits of cinnamon are due to both its insulin‐like and insulin‐independent effects (Supported in part by USDA‐ARS Human Nutrition Research Program).
SNPs at IL‐2 and IL‐10 genes are associated with respiratory infection in the elderly and may modulate the effect of vitamin E on lower respiratory infections in elderly women
Authors: Belisle Sarah E. , Hamer Davidson H. , Leka Lynette S. , Dallal Gerard E. , Delgado‐Lista Javier , Fine Basil C. , Jacques Paul F. , Ordovas Jose M. , Meydani Simin Nikbin
Published: June 2021

Vitamin E (E) supplementation has been suggested as a preventative measure against respiratory infections (RI) in the elderly. Previously, we showed that E reduced RI in some but not all nursing home residents. The efficacy of E supplementation may depend on individual factors, including specific single nucleotide polymorphisms (SNPs) at immunoregulatory genes. We examined whether a E‐gene interaction might explain individual responses to E for protection from RI using data from a previous E intervention study in elderly nursing home residents. The genotype of common SNPs at IL‐1β, IL‐2, IL‐6, IL‐10, TNF‐α, and IFN‐γ genes were determined. The effect of E on lower respiratory infections (LRIs) depended on gender and IL‐10 ‐819G>A (P=0.03 for interaction). Further, IL‐2 ‐330A>C (P=0.02 for upper respiratory infection (URI)), IL‐10 ‐819G>A (P=0.08 for URI) and IL‐10 ‐1082C>T (P<0.001 for LRI in men) were associated with RI independent of E. Genetics and gender should be considered in future studies of E and LRI. SNPs at cytokine genes may be linked to individual risk of RI in the elderly. Supported by NIA NIH grant 5R01‐AG013975, USDA agreement 58‐1950‐7‐707, and a DSM Nutritional Products, Inc. scholarship. CIBEROBN is an initiative of ISCIII.
Green tea EGCG suppresses T cell proliferation by impairing IL‐2/IL‐2R signaling leading to inhibition of cell cycle
Authors: Pae Munkyong , Meydani Simin Nikbin , Meydani Mohsen , Shang Fu , Wu Dayong
Published: June 2021

Epigallocatechin‐3‐gallate (EGCG), an active ingredient of green tea, has been indicated to have various health benefits. However, little is known about the effect of EGCG on immune function. In this study, we investigated the effect of in vitro EGCG supplementation at physiologically relevant levels on T cell proliferation and its underlying mechanism. Purified T cells were isolated from the spleens of male C57BL/6 mice and stimulated with anti‐CD3/CD28 in the presence of EGCG (2.5‐15 μM). EGCG dose‐dependently inhibited T cell proliferation and cell division, which was more pronounced in CD4+ than in CD8+ T cells. The EGCG‐induced inhibition of T cell division was due to cell cycle arrest as fewer cells entered S‐G2‐M phase. Since IL‐2 is a key factor for T cell proliferation, we determined the effect of EGCG on IL‐2 and IL‐2 receptor (IL‐2R, CD25) levels. Surprisingly, EGCG increased IL‐2 secretion in total T cells. Intracellular staining for IL‐2 indicates that EGCG increased percentage of IL‐2+ cells in CD8+ but not CD4+ population. Furthermore, EGCG reduced the mean intensity of IL‐2 in both CD8+ and CD4+ cells. EGCG did not significantly affect the percentage of CD25+ (IL‐2R) cells but reduced the IL‐2R expression per cell. This suggests that EGCG may inhibit T cell proliferation by inducing cell cycle arrest, which is in turn due to impaired IL‐2/IL‐2R signaling. Increased extracellular IL‐2 might result from decreased internalization and utilization due to less expression of IL‐2R. T cell‐suppressive effect of EGCG might be utilized to ameliorate T cell‐mediated autoimmune diseases.

Grant Funding Source

USDA #58‐1950‐7‐707
Dietary curcumin and limonin suppress murine CD4+ T‐cell activation
Authors: Kim Wooki , Fan Yang‐Yi , Smith Roger , Patil Bhimanagouda S. , Jayaprakasha G. K. , McMurray David N. , Chapkin Robert S.
Published: June 2021

Phytochemicals may reduce chronic inflammation and cancer risk, in part, by modulating T‐cell nuclear factor kappa B (NF‐κB) activation. Therefore, we examined the effects of curcumin (C) and limonin (L) feeding on the NF‐κB pathway in CD4+ T‐cells. DO11.10 transgenic mice were fed diets containing 1% curcumin or 0.02% limonin, in the presence of n‐6 PUFA (corn oil) or with n‐3 PUFA (fish oil) for two weeks, followed by splenic CD4+ T‐cell isolation and stimulation with OVA323‐339 peptide and antigen presenting cells from mice fed the conventional rodent chow. Both C and L diets suppressed (P<0.05) NF‐κB p65 nuclear translocation in activated CD4+ T‐cells. In contrast, AP‐1 c‐Jun and NFATc1 were not affected relative to control (corn oil, no C or L). CD4+ T‐cell proliferation in response to either mitogenic anti‐CD3/28 mAbs or antigenic stimulation by OVA peptide was also suppressed (P<0.05) by C as assessed by CFSE staining. In contrast, IL‐2 production was not directly associated with NF‐κB status. Interestingly, dietary combination with fish oil enhanced the suppressive effects (P<0.05) of C or L with respect to CD4+ T‐cell proliferation in response to anti‐CD3/28 mAbs. These results suggest that combination chemotherapy (fish oil + curcumin or limonin) may favorably modulate CD4+ T‐cell mediated inflammation. (Supported by NIH DK71707, CA59034, CA129444, P30ES09106, and USDA 2006‐34402‐17121)
Vitamin D3 content of fortified yogurt and milk as determined for the USDA National Food and Nutrient Analysis Program (NFNAP)
Authors: Holden Joanne M. , Patterson Kristine Y , Exler Jacob , Horst Ronald L , Lemar Linda E , Phillips Katherine M.
Published: June 2021

In response to recent interest in vitamin D composition of foods, USDA‐NDL is updating and expanding data in the National Nutrient Database for Standard Reference. In 2007, the USDA sampled vitamin D3 fortified yogurt and milk from 12 and 24 supermarkets, respectively, selected from a nationwide statistical sampling. The four types of yogurt collected were from two national manufacturers while the milk samples were processed in dairies in the vicinity of the supermarkets. The samples along with QC material were analyzed for vitamin D3 by HPLC with UV detection. All of the yogurt samples met the label claim (15% or 20%DV). Three of the four types of yogurt had vitamin D3 values at 100 ‐ 150% of label claim, with the fourth type above 150% but not exceeding 175%. Each serving of yogurt had 1.6 to 3.0 µg vitamin D3. Five types of fortified milk were collected; skim, 1%, 2%, whole and 1% chocolate. All the products should provide 25%DV of vitamin D3 / 8 oz serving (2.6 µg). Actual vitamin content varied from zero to 47%DV, nearly double the label claim. Overall, about 77% of the products fell within or very close to the expected 100 ‐ 150% of label, however, about 17% of the values were lower than 100%. The vitamin D content of milk was more variable than yogurt. Accurate, current data for vitamin D are important to the assessment of intake by the US population.

Support by USDA, NIH #Y1CN5010 and the Beverage Institute for Health & Wellness (Coca‐Cola)
Adherence to 2005 Dietary Guidelines for Americans was associated with reduced progression of coronary‐artery atherosclerosis in women with established coronary artery disease
Authors: Imamura Fumiaki , Jacques Paul F , Herrington David M , Dallal Gerard E , Lichtenstein Alice H
Published: June 2021

Criterion‐based dietary indices generally have an untested assumption that all index components have equal health contributions. This assumption was tested for the association between adherence to 2005 Dietary Guidelines for Americans (DGA) and atherosclerosis progression, using data from 225 postmenopausal women with established heart disease in the Estrogen and Atherosclerosis Progression Trial conducted between 1996 and 2000. At the baseline and endpoint, diameters of coronary arteries were measured by quantitative angiography. Adherence to the key DGA recommendations was measured by the DGA Adherence Index (DGAI), where each component was weighted equally, and the modified DGAI (wDGAI), where each component was weighted by its relation to atherosclerosis progression. By mixed model regression analyses, the DGAI was found not associated with atherosclerosis progression (p=0.44). In contrast, wDGAI was inversely associated; one standard deviation difference of wDGAI was related to 0.049 mm less narrowing of coronary arteries (p=0.004). Adherence to the DGA recommendations assigned differential weights based on their relationships with the outcome was significantly associated with reduced atherosclerosis progression. Assuming equity of associations of all DGA recommendations with health outcomes may limit our ability to identify relationships of the DGA with chronic disease prevention.

Grant Funding Source

U.S. Department of Agriculture, Agricultural Research Service (No. 58‐1950‐7‐707)
Zinc status affects DNA damage and oxidative stress in healthy adult men
Authors: Song Yang , Chung Carolyn S , Bruno Richard S , Traber Maret G , Brown Kenneth H , King Janet C
Published: June 2021

Zinc plays an important role in the maintenance of DNA integrity and antioxidant defense in cells. Our lab has shown in rats that zinc depletion increases DNA damage and perturbs oxidative stress. In the current human dietary intervention study, we aimed to evaluate the effects of zinc depletion and repletion on DNA damage, oxidative stress and antioxidant defenses in humans. Total 9 healthy men with daily zinc intake >11mg/d were recruited. Subjects underwent three consecutive dietary periods: baseline (2 wks, 11mg Zn/d), zinc‐depletion (0.6 mg Zn/d for 1 wk and 4 mg Zn/d for 5 wks), and zinc‐repletion (4 wks, 11 mg Zn/d). DNA damage in peripheral blood cells, plasma oxidative stress and antioxidant defenses were assessed at different time points. Human dietary zinc depletion caused a significant increase in DNA strand breaks in peripheral blood cells (P<0.05), but did not alter lipid peroxidation, as indicated by plasma F2‐isoprostanes. The increased DNA strand breaks were reduced to baseline levels following zinc repletion (P<0.05). Although plasma tocopherol levels significantly increased after zinc depletion, plasma total antioxidant capacity and erythrocyte SOD were not changed. This is first study that demonstrates increases in DNA with human zinc depletion and suggests that zinc may be essential for maintaining DNA integrity in humans. This work is supported by the NCBA (KHB) and USDA 2005‐35200‐15439 (EH).

Grant Funding Source

NCBA (KHB) and USDA 2005‐35200‐15439
The Abundance and Activation of mTORC1 Regulators in Skeletal Muscle of Neonatal Pigs are Modulated by Insulin, Amino Acids, and Age
Authors: Suryawan Agus , Orellana Renán , Nguyen Hanh , Davis Teresa
Published: June 2021

Previously we demonstrated that the insulin (INS) and amino acid (AA) ‐induced activation of the mammalian target of rapamycin complex 1 (mTORC1) is developmentally regulated in neonatal pigs. This study aimed to determine the effects of the post‐prandial rise in INS and AA on the activation and abundance of mTORC1 regulators and how this is modified by development. Overnight fasted 6‐ and 26‐day‐old pigs were infused for 2 h with saline (Control) or with INS or AA to achieve fed levels. Phosphorylation of Proline‐rich Akt Substrate of 40 kDa (PRAS40) by protein kinase B alleviates the inhibitory effect of PRAS40 on mTORC1 activation. INS, but not AA, enhanced PRAS40 phosphorylation and this effect was higher in younger pigs. AMP‐activated protein kinase inactivates mTORC1 by phosphorylating raptor. Neither INS, AA, or age altered the phosphorylation of raptor. Rheb, vacuolar protein sorting mutant 34 (Vps34), and ras‐related GTP‐binding protein B (Rag B) are activators while FK506‐binding protein 38 (FKBP38) is an inhibitor of mTORC1. Neither INS, AA, or age altered the abundance of rheb, Vps34, or FKBP38. Although INS and AA had no effect, the abundance of Rag B was greater in young than in older pigs. Our results suggest that the higher mTORC1 activation in young than in older pigs is in part due to regulation by PRAS40 and the Rag GTPases. (Supported by NIH grants AR‐44474 and USDA/ARS 6250510000‐33).
The Association between Alcohol Consumption and Diabetes Risk is Independent of Dietary Pattern
Authors: Imamura Fumiaki , Lichtenstein Alice H , Meigs James B , Dallal Gerard E , Jacques Paul F
Published: June 2021

The ability to interpret epidemiologic relations of diet and disease risk is limited due to potential residual confounding by correlated dietary components. Assessing dietary patterns by factor analysis or partial least squares may overcome this limitation. To examine confounding by dietary pattern as well as standard risk factors and selected nutrients, the longitudinal association of alcohol consumption with 7‐year risk of type 2 diabetes mellitus (T2DM) was analyzed by Cox‐proportional hazard model in 2,879 healthy adults enrolled in Framingham Offspring Study. After adjusting for standard risk factors, consumers of ≥9.0 drink/week had a significantly lower risk of T2DM compared to abstainers (hazard ratio [HR]: 0.47; 95% confidence interval [CI]: 0.27, 0.81). Adjusting for selected nutrients little impacted the HR, whereas adjusting for dietary pattern variables by factor analysis significantly shifted the HR away from null (HR: 0.33, 95% CI: 0.17, 0.64) by 40.0% (95% CI: 16.8, 36.4%, P=0.002). Dietary pattern variables by partial least squares showed the similar results. Therefore, the observed inverse association, consistent with past studies, was confounded by dietary patterns, which was not corrected by adjusting for individual nutrients. The data suggest alcohol intake, and not dietary patterns associated with alcohol intake, is responsible for the observed inverse association with T2DM risk.

Grant Funding Source

U.S. Department of Agriculture, Agricultural Research Service (No. 58‐1950‐7‐707)
Long‐chain polyunsaturated fatty acids inhibit infectivity of Apicomplexan parasites
Authors: Lee Jennifer Hanbyul , McAllister Milton M. , Kuhlenschmidt Theresa B. , Kuhlenschmidt Mark S.
Published: June 2021

Cryptosporidium parvum, Toxoplasma gondii, and Plasmodium gallinaceum are parasitic pathogens of the phylum Apicomplexa. These parasites commonly infect a variety of vertebrate species and can produce life‐threatening diseases, especially in the immunocompromised. Previously, we have shown that certain, colostrum‐derived, long‐chain polyunsaturated fatty acids (L‐PUFA) inhibit C. parvum sporozoite‐host‐cell invasion. In the studies reported here, we show linolenate or oleate but not elaidic acid, the trans isomer of oleate, inhibits both C. parvum sporozoite and T. gondii tachyzoite infectivity in vitro. Similarly, linolenate also blocked in vivo infectivity of T. gondii tachyzoites and P. gallinaceum sporozoites. Preliminary mechanistic studies suggest linolenate or oleate but not elaidic acid, block gliding motility of T. gondii tachyzoites and C. parvum sporozoites. While these data suggest that certain L‐PUFAs represent possible therapeutic nutriceuticals, they clearly serve as useful reagents to probe what may be a common mechanism of host‐cell invasion by Apicomplexan parasites.

Grant Funding Source

USDA
Wild Blueberry (V. angustifolium) Consumption Alters the Composition and Structure of Glycosaminoglycans (GAGs) in Spontaneously Hypertensive Rats (SHR)
Authors: Kristo Aleksandra S. , Malavaki Christina J. , Lamari Fotini N. , Karamanos Nikos K. , Klimis‐Zacas Dorothy J.
Published: June 2021

The effect of wild blueberries on the composition and structure of GAGs was examined in twenty, 12 week‐old SHRs, fed a control (C) or blueberry‐enriched diet (B) for nine weeks. Thoracic aortae were defatted with methanol/chloroform solution (1:1 v/v), digested with pronase and assayed for total sulfated GAGs and uronic acid content. Aortic specimens from each diet group were pooled together and fractionated on a DEAE‐Sephacel column with a NaCl linear gradient. Identification and characterization of pooled uronic acid‐positive fractions was determined with cellulose acetate electrophoresis, treatment with specific GAG lyases and high performance capillary electrophoresis. A significant decrease in total sulfated GAGs was documented in the SHR‐B, 12.58 ± 0.20 vs. 13.72 ± 0.20 µg/mg aorta in the SHR‐C, p=0.05. Three different GAG populations were identified; hyaluronic acid (HA), heparan sulfate (HS) and galactosaminoglycans (GalAGs). There was a 25% and 10% increase in HA and HS, respectively, and a 13% decrease in GalAGs in the SHR‐B as compared to SHR‐C group. The changes in the SHR aortic GAG profile produced by wild blueberry consumption may alter cellular signal transduction pathways and subsequently affect the biomechanical milieu of the vascular endothelium.

This work was supported by a USDA/CSREES (WBANA) Grant and a Greek State Scholarship Foundation Fellowship (IKY).

Grant Funding Source

WBANA, IKY
Effects of diets naturally high or low in conjugated linoleic acid on measures of health in women
Authors: Brown Andrew William , Trenkle Allen H , Beitz Donald C
Published: June 2021

Conjugated linoleic acid (CLA) consumption purportedly alters body composition, glucose tolerance, lipoprotein distributions, and other health markers, though reports are often contradictory or inconclusive. The present study aimed to determine if CLA in a natural food matrix alters selected markers of health by presenting isocaloric, nutritionally complete diets containing 31% energy from lipid, 13% energy from protein, and 54% energy from carbohydrates that contained either products naturally high in CLA (1.17 g/d) from pasture‐fed cattle or naturally low in CLA (0.35 g/d) from grain‐fed cattle to 18 generally healthy women age 20‐39 y in a parallel arm study. Incorporation of products of pasture‐fed origin enriched with CLA resulted in no differences in insulin sensitivity, body composition, circulating lipids, nor other measured disease risk factors as compared with the commercial, low CLA diet. We conclude that a diet naturally enriched with a 3.5 fold increase in CLA by incorporation of products from pasture‐fed cattle added no measured health benefits when compared to the low CLA diet.

Grant Funding Source

USDA‐CSREES no. 2001‐38640‐10270
An integrated economic and education intervention (the ENAM project) decreased household food insecurity in rural Ghana
Authors: Harding Kimberly B , Marquis Grace S , Colecraft Esi K , Lartey Anna , Sakyi‐Dawson Owuraku , Ahunu B K , Reddy Manju B , Jensen Helen H , Butler Lorna M , Lonergan Elisabeth
Published: June 2021

Interventions that enhance women's incomes and nutrition knowledge are likely to improve household food security through increased economic contribution to food expenditures and better nutrition practices. From May 2006 to September 2007, the ENAM project provided nutrition education and enterprise development services for 180 caregivers of young children in three ecological zones of Ghana, with the goal of improving household food security and child nutrition. A control sample of 287 households were also recruited. At baseline, almost three‐quarters of households expressed concern about their inability to meet their food needs and about half of adults and children were reported to consume less food than desired because of poverty. Locale, household size and type, and income were associated independently with food security in the past month (p<0.05). The logistic regression model demonstrated that, compared to control households, there was about a 50% decrease in the risk of food insecurity among intervention households at the final intervention time point (p<0.05). An integrated intervention that addresses barriers to availability, accessibility, and utilization of ASF can reduce household food insecurity. This is a collaborative research effort of ENAM researchers with support through the GL‐CRSP, funded in part by USAID, Grant # PCE‐G‐00‐98‐00036‐00.
Mitochondrially targeted vitamin E decreases diet‐induced fat deposition in C57BL/6 mice through normalizing mitochondrial oxidative stress and function
Authors: Mao Gaowei , Kraus George , Spurlock Michael , Beitz Donald
Published: June 2021

The objective of this study was to evaluate the effect of a novel modified vitamin E (MitoVit E) that can penetrate mitochondria for minimization of reactive oxygen species on mitochondrial function, animal growth and obesity. Sixty‐four male 9 weeks old C57BL/6J mice, after being fed a high fat diet (60 kcal% from fat) for 5 weeks, were assigned randomly to 4 groups. Group I and II were fed a low fat diet (10 kcal% from fat) and a high sucrose diet (20 kcal% from sucrose), respectively, with each group gavaged with drug vehicle (0.9% NaCl solution); Group III and IV were fed the same diet of Group I and II, respectively, with each group gavaged 40 mg MitoVit E/kg body weight. Gavage was conducted every other day for 5 weeks. We showed that high sucrose diet significantly resulted in increased inguinal (p=0.0225) and epididymal (p=0.0004) fat deposition and increased urinary isoprostane (p=0.0006), all of which were normalized by MitoVit E. MitoVit E significantly decreased H2O2 production (p<0.05) and increased ATP production (p<0.05) from the isolated mitochondria in the liver but not in the muscle. Superoxide dismutase in the plasma was also improved by the MitoVit E (p=0.0143). These data indicated that oxidative stress resulting from high fat/high sucrose diets contributed to mitochondrial dysfunction and fat deposition, which, however, can be improved by mitochondrially targeted MitoVit E.

Funding: USDA

Grant Funding Source

USDA
Deoxycholic acid and selenium metabolite methylselenol exert common and distinct effects on cell cycle, apoptosis, and MAP kinase pathway in HCT116 human colon cancer cells
Authors: Zeng Huawei , Botnen James H , Briske‐Anderson Mary
Published: June 2021

The cell growth inhibition induced by bile acid deoxycholic acid (DCA) may cause compensatory hyperproliferation of colonic epithelial cells, and consequently increase colon cancer risk. On the other hand, there is increasing evidence for the efficacy of certain forms of selenium (Se) as anticancer nutrients. Methylselenol has been hypothesized to be a critical Se metabolite for anticancer activity in vivo. In this study, we demonstrated that, both DCA (75 ‐ 300 μmol/L) and submicromolar methylselenol, inhibited colon cancer cell proliferation by up to 64% and 63%, respectively. In addition, DCA and methylselenol each increased colon cancer cell apoptosis rate by up to 2‐fold. Cell cycle analyses revealed that DCA induced an increase in only the G1 fraction with a concomitant drop in G2 and S‐phase; in contrast, methylselenol led to an increase in the G1 and G2 fractions with a concomitant drop only in the S‐phase. Although both DCA and methylselenol significantly promoted apoptosis and inhibited cell growth, examination of mitogen‐activated protein kinase (MAPK) pathway activation showed that DCA, but not methylselenol, induced SAPK/JNK1/2, p38 MAPK, ERK1/2 activation. Thus, our data provide, for the first time, the molecular basis for opposite effects of methylselenol and DCA on colon tumorigenesis. Research supported by USDA/ARS.
Will lutein supplementation of lactating women affect the lutein content of milk and the lutein content of maternal and preterm infant plasma?
Authors: Connor Sonja L. , Bezzerides Eugenia , Pillers De‐Ann , Connor William E.
Published: June 2021

Breast milk and blood were obtained from 38 lactating women. Twenty one of these women took a lutein supplement of 12 mg per day weeks five through eight of lactation. Blood was also obtained from five preterm infants whose mothers took the supplement. Breast milk and plasma carotenoids were analyzed by high‐pressure liquid chromatography. Maternal plasma lutein increased 241 percent after four weeks of the lutein supplement (10.6 ± 7.3 to 36.1 ± 13.1 mcg/dL (p=0.000). By four weeks post supplement, plasma lutein returned to baseline (10.4 ± 3.3 mcg/dL). Non‐supplemented women had no change in plasma lutein (10.1 ± 5.4 mcg/dL [4 wks], 9.8 ± 5.1 mcg/dL [8 wks], 10.3 ± 5.6 mcg/dL [12 wks]). Breast milk lutein content in non‐supplemented women was 4.3 ± 3.4 mcg/dL (4 wks), 3.8 ± 3.6 mcg/dL (8 wks) and 4.5 ± 5.1 mcg/dL (12 wks). Lutein‐supplemented women had breast milk lutein increase from 3.1 ± 2.7 to 11.0 ± 7.7, after only one week, to 14.5 ± 16.4 mcg/dL (p=0.011) after four weeks and returned to baseline four weeks post supplement (3.8 ± 4.3 mcg/dL). Plasma lutein increased 535 percent (1.7 ± 1.1 to 10.8 ± 1.8 mcg/dL) in the five breast‐fed preterm infants after supplementation of their mothers with lutein for four weeks. Giving a lutein supplement to lactating women produced a dramatic increase in the lutein content of their plasma and breast milk and in the lutein content of preterm infant plasma. USDA grant and Wyeth Pharmaceuticals
BLUEBERRY JUICE AND ANTHOCYANINS MODULATE OBESITY, LEPTIN AND BETA CELL FUNCTION IN MICE FED A HIGH FAT DIET
Authors: Prior Ronald L. , Wilkes S. , Rogers T. , Khanal R. C. , Wu X. , Howard L. R.
Published: June 2021

Consumption of purified blueberry (BB) anthocyanins (ACNs) but not whole BB in the diet has been shown to prevent the development of obesity in mice fed high fat diets (JAFC 56:647, 2008). Our objective was to determine whether BB juice (BBJ) retained the factors responsible for the obesigenic effects of whole BB. Male C57BL/6 mice were assigned to 1 of 6 diet groups: 1) 10% of kcal from fat (LF); 2) a high fat diet [45% kcal from fat (HF45)]; 3) LF diet plus purified ACNs from BBs (0.2 mg total ACNs/mL) in drinking water; 4) HF45 diet plus BB ACNs; 5) LF diet plus BBJ in place of drinking water; 6) HF45 diet plus BBJ. Mice were fed for a total of 72 days. In mice fed the HF45 diet, fat mass as determined by MRI and weights of epididymal and retroperitoneal fat were reduced by 26‐28% (p<0.01) with consumption of purified ACNs and 8‐12% in those consuming BBJ. Serum leptin was increased in mice fed HF45 diet relative to LF control and was decreased to control levels in mice fed blueberry ACNs or BBJ (p<0.003). Pancreatic beta cell function (beta cell Homeostasis Assessment Score) was decreased and serum glucose was increased with HF45 feeding and both returned to normal with HF45 + BB ACNs in the drinking water but not with BBJ. Purified BB ACNs in the drinking water were more effective in preventing obesity in HF45 fed mice compared to BBJ containing an equivalent quantity of ACNs. Supported by: USDA‐NRI Grant 2005‐35503‐15409.
Integrating economic and educational intervention activities in the ENAM project leads to improved child nutritional status in rural Ghana
Authors: Marquis Grace S , Harding Kimberly B , Colecraft Esi K , Lartey Anna , Sakyi‐Dawson Owuraku , Ahunu B K , Reddy Manju B , Jensen Helen H , Butler Lorna M , Lonergan Elisabeth
Published: June 2021

Animal source foods (ASF) are excellent sources of micronutrients that are essential for growth and development of young children. In sub‐Saharan Africa, children's typical diets are based on starchy staples and little ASF, contributing to high rates of child malnutrition. The Enhancing Child Nutrition through Animal Source Food Management (ENAM) project identified constraints to the availability, accessibility, and utilization of ASF in the diets of young children (2‐5 y) in Ghana. Based on the identified constraints, an intervention involving micro‐credit and nutrition and entrepreneurial education with 180 caregivers of young children was implemented in six communities in three agro‐ecological zones; 287 households in matched communities were chosen as controls. Child height and weight were measured at baseline and every 4 mo. Children from intervention households showed a greater increase in height‐for‐age Z‐score over the course of the 16‐month study than children from control households (0.3 ± 0.6 vs. 0.1 ± 0.4 Z‐score; p<0.05). There were no between‐group differences in weight‐for‐height or weight‐for‐age Z‐scores. An integrated intervention that addresses barriers to availability, accessibility, and utilization of ASF can improve child nutrition. Funding: GL‐CRSP, funded in part by USAID, Grant # PCE‐G‐00‐98‐00036‐00.
Increases in caregivers' contributions to household food and non‐food expenditures did not affect child outcomes in the ENAM project
Authors: Colecraft Esi K , Marquis Grace S , Lartey Anna , Sakyi‐Dawson Owuraku , Ahunu B K , Butler Lorna M , Reddy Manju B , Jensen Helen H , Lonergan Elisabeth
Published: June 2021

Interventions that enhance women's incomes are likely to improve child nutrition outcomes. This association may be reduced if women increase their economic contribution primarily to household non‐food rather than food expenditures. The ENAM project is a nutrition education and enterprise development intervention aimed at improving caregiver incomes and children's animal source food (ASF) intakes. We assessed caregivers' contributions to household food and non‐food expenditures and their differential effects on children's ASF diversity and growth. 179 intervention and 287 control caregivers were interviewed about their household expenditures at 4 quarterly time points. Children's ASF intakes and anthropometry were also recorded. There were no between‐group differences at baseline. At the final time point, compared to control households, the intervention households had higher percentage increases in caregiver contribution to food (52.5% ± 40.4% vs 66.2% ± 33.1%; P<0.001) and non‐food (51.8% ± 34.1% vs. 63.9% ± 29.2%; p<0.05) expenditures. However, neither increased caregivers' contributions to food nor non‐food expenditures affected children's ASF diversity or anthropometry. An increase in the proportion of household expenditures coming from caregivers did not affect child nutritional outcomes. Funding: GL‐CRSP, funded in part by USAID, Grant # PCE‐G‐00‐98‐00036‐00.
ISOLATION AND QUANTITATION OF ANTI‐INFLAMMATORY COMPOUNDS IN GARLIC (ALLIUM SATIVUM).
Authors: Park Jae B
Published: June 2021

Allium sativum (garlic) is a well‐known medicinal herb containing significant anti‐hypertensive, anti‐cholesterol, anti‐oxidant and anti‐inflammatory activities. In this study, anti‐inflammatory phytochemicals were isolated from crude garlic by sequential extraction with methanol and hexane, and the fractionation using silica‐gel chromatography. The fractions were tested for the inhibition of COX enzymes (I and II) and the production of thromboxane B2 (thromboxane A2 derivative) because these enzymes and their metabolites are important in inflammatory processes. Those fractions with the highest COX‐inhibitory activity were further purified using HPLC, and again assayed for COX‐inhibitory activity. The identity of the compounds was analyzed using LC‐MS and MS‐MS. In addition, the isolated compounds were quantified using a HPLC method, and their effects on inflammatory bio‐markers (e.g., COX enzymes, thromboxane B2, others) were determined in vitro and in vivo. These compounds are likely to have potential long‐term health effects on inflammation and other chronic diseases. This study was funded by USDA project 1235‐52530‐051‐00D.
Potential predictive markers for breast cancer aggressiveness and invasiveness
Authors: Wang Wan , Wang Qiuyue , Gao Ling , Gone Pengtao , Zhang Xichen , Man Yan‐gao
Published: June 2021

Clinical follow‐up studies have shown that only 10‐30% of untreated in situ breast carcinoma progress to invasion during patients' lifetime. None of the current approaches, however, could predict tumor invasiveness or identify the specific individuals at greater risk for invasive lesions. As myoepithelial (ME) cells are the sole source of several tumor suppressors and the disruption of ME cell layer is a pre‐requisite for tumor invasion, our recent studies compared the expression profile of tumor suppressors and the physical integrity of ME cell layers in pregnancy‐associated breast cancer (PABC) and inflammatory breast cancers (IBC), which have the highest degree of aggressiveness and invasiveness, with those of clinically idle breast cancers. Compared to their counterparts, the ME cell layers in pre‐invasive PABC and IBC have several unique properties: [1] significantly lower expression of tumor suppressors, especially WT‐1, [2] a significantly higher frequency of focal disruptions, and [3] a significantly higher infiltration of immunoreactive cells. Our studies suggest that the expression of tumor suppressors and the physical integrity in ME cell layers may have significant predictive value for breast tumor aggressiveness and invasiveness ((Supported in part by grant 2006CB910505 from the Ministry of Chinese Science and Technology Department, and by grants DAMD17‐01‐1‐0129, DAMD1701‐01‐1‐0130, PC051308 from Congressionally Directed Medical Research Programs, and grant BCTR0706983 from The Susan G. Komen Breast Cancer Foundation).
Undergraduate Bioinformatics: Evaluating the effectiveness of a short lab module compared to a semester long course
Authors: Matejka Stephanie L , Soundararajan Madhavan , Bailey Cheryl
Published: June 2021

At large universities and some smaller institutions, not all undergraduate biochemistry majors are involved in a research project. This is unfortunate, since the scientific method and research process provide excellent learning opportunities. The Joint Genome Institute (JGI) has launched a pilot project where students annotate microbial genomes and fundamental metabolic pathways. The JGI integrated microbial genome platform, IMG/ACT, provides an interface for students to analyze genes and pathways and report their findings. This project emulates the research experience and results in new knowledge available for other scientists. This year is the second offering of a specialized semester‐long bioinformatics course that focuses on this initiative. We have also tested the effectiveness of broadening the experience by introducing a short annotation module into an existing biochemistry lab course. Both implementation types can be evaluated through surveys and confidence of project results. This compares the effectiveness of both the semester long course and the shorter lab module.

Funded by the Department of Energy.
Use of chemical genetics to study zebrafish gut development
Authors: Devlin Ed , Nascone‐Yoder Nanette , Dush Mike , Bloom Stephanie
Published: June 2021

The signaling pathway associated with the formation and looping of the early Zebrafish gut is poorly understood. The present study uses a chemical genetics approach to examine aspects of the signaling pathway thought to be involved in the process of gut looping in the Zebrafish. A chemical genetics approach is one that uses small molecules known to modulate the activities of specific proteins in a signaling pathway in order to study their role tissue or organ morphogenseis. Compounds thought to be involved in gut looping used in the present study included rho kinase III inhibitor (rockout), blebbistatin which acts as a reversible inhibitor of non‐muscle myosin II, retinoic acid (RA) and DEAB. Effective maximum working concentrations that allowed for hatching of Zebrafish larvae were determined to be 5 uM DEAB, 0.5 uM RA, 1‐2 uM Blebbistatin and 10 uM Rockout. Gut looping was visualized in 30‐32 hpf embryos utilizing WISH for Fox A3 that recognizes early endoderm tissue of the gut tube and with Zebrafish gut‐GFP mutants (Tg(XlEef1a1:GFP). In addition, IHC staining for laminin, pan‐cadherin, B‐catinin, PKC and smooth muscle actin was utilized for visualization of the polarity, orientation and movement of the endodermal and mesodermal cell layers of the early and later stage Zebrafish gut tube. WISH and gut‐GFP mutant experiments demonstrated that 10 uM rockout but not 2 uM blebbistatin resulted in an inhibition of gut looping in 30‐32 hour old Zebrafish embryos. This work was performed in the lab of Dr. Nannette Nascone‐Yoder and supported by NSF award # 0642012.