bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Fast Food Consumption on Dietary Intake and Likelihood of Meeting MyPyramid Recommendations in Adults: Results from What We Eat in America, NHANES, 2003–2004
Authors: Sebastian Rhonda , Enns Cecilia Wilkinson , Goldman Joseph D. , Moshfegh Alanna J
Published: June 2021

The purpose of this study was to determine the effect of fast food (FF) consumption on food and nutrient intakes and likelihood of meeting recommendations outlined in USDA's MyPyramid Food Guidance System. Adults 19–50 years of age (n=2,160) who completed two 24‐hour dietary recalls in the What We Eat in America, NHANES, 2003–2004 were classified as FF consumers or non‐consumers. Fast food consumers were divided into tertiles based upon the proportion of 2‐day energy intake derived from FF. Regression procedures adjusting for confounding factors were used to detect associations between FF consumption and food and nutrient intakes and to predict the odds of meeting MyPyramid recommendations by FF consumption level. Increasing tertiles of FF intake were associated with decreasing intake of most nutrients analyzed, and, for men only, decreasing intakes from the MyPyramid fruit, meat/beans, and oils groups (p<0.01). Compared to non‐consumers, FF consumers had higher intakes of total energy, fat, saturated fat, and discretionary energy. Except for the oils group for men and the grains group for women, FF consumption did not affect the likelihood of meeting recommendations for MyPyramid components. Overall, although fast food consumption was associated with higher intakes of energy, it did not improve food and nutrient intakes or enhance the likelihood of meeting most MyPyramid recommendations. Funding source for this study is USDA.
Vitamin B6 status of Ghanaian infants
Authors: Dewey K G , Yang Z , Miller J W , Adu‐Afarwuah S , Lartey A , Brown K H , Briend A , Zlotkin S
Published: June 2021

Vitamin B6 intake from complementary foods is often low, but data are lacking on prevalence of B6 deficiency. We measured erythrocyte pyridoxal 5′‐phosphate concentration (EPLP) by HPLC in Ghanaian infants enrolled in an efficacy trial of Sprinkles (SP), Nutritabs (NT) and Nutributter (NB) added to home‐prepared complementary foods. SP contained Fe, Zn, vitamins A and C and folic acid; NT and NB contained these and several other micronutrients including 0.3 mg of B6. Infants (n = 313) were randomly assigned to receive SP, NT or NB daily from 6 to 12 mo of age. Non‐Intervention (NI) infants (n=96) were assessed at 12 mo only. EPLP was <51.6 nmol/L (the 10th percentile for Finnish breastfed infants) in 57.5% at 6 mo and 56.5% at 12 mo. However, there was no significant impact of NT or NB on EPLP at 12 mo [mean (SD): SP 53.2 (36.7), NT 53.5 (21.3), NB 52.0 (22.1), NI 59.9 (46.4)]. EPLP was inversely related to number of siblings at 6 (r=−0.20, p<0.01) and 12 mo (r=−0.13, p<0.05) and to socioeconomic status at 12 mo (r=−0.13, p<0.05). EPLP at 6 mo was not associated with subsequent growth or motor development after controlling for intervention group and potential confounders. Although B6 deficiency appears to be common in this population, provision of one RDA of B6 daily for 6 mo did not alter EPLP at 12 mo. Further research is needed to evaluate potential functional consequences of low EPLP.

Supported by Nestle Foundation and USAID.
Effect of antioxidants on the oxidative stress in cataract patients
Authors: Yoshida Yasukazu , Li Lei , Duker Jay , Niki Etsuo , Russell Robert M , Yeum Kyung‐Jin
Published: June 2021

To determine the relationship between oxidative stress and eye disease, and the impact of antioxidant supplementation, forty subjects (50–70 y, F25, M15) were enrolled in a double blinded randomized study. Subjects were randomized to receive either 1) lutein (12 mg) or 2) lutein (12 mg) + green tea extract (200 mg) for 16 wks, and underwent eye examinations before (wk 0) and after supplementation (wk 16). Lutein is present in human lens and macula. Blood samples were collected at 0, 4, 8, 12 & 16 wks and analyzed for antioxidants by HPLC systems, total antioxidant performance by fluorometry, malondialdehyde by an HPLC with a fluorescent detector, 8‐iso‐prostaglandin‐F2α & total hydroxyoctadecadienoic acid (tHODE) by a GC‐MS. Twenty seven subjects (F17, M10) were determined to have cataract at wk 0. In these subjects, tHODE level was significantly higher than the healthy subjects (P<0.05) at wk 0. Lutein supplementation resulted in a significant increase in plasma lutein in cataract patients as early as wk 4 & throughout the study period without any significant change in other antioxidants or oxidative stress biomarkers. Our data suggest that cataract patients were under high oxidative stress and their plasma lutein concentration can be significantly increased by a daily dose of 12 mg lutein, which did not affect overall antioxidant capacity/oxidative status in plasma. (Supported by NEI R03EY015674 & USDA #581950‐7‐707)
Gastrointestinal Structural and Functional Responses to Rotavirus Infection in the Neonatal Piglet
Authors: Mathai Rose Ann , Andres Aline , Helregel Beth A. , Kuhlenschmidt Theresa B. , Kuhlenschmidt Mark S. , Donovan Sharon M.
Published: June 2021

Rotavirus (RV) infection is a primary cause of diarrheal diseases worldwide. RV infects villus epithelial cells, causing villus blunting and malabsorptive diarrhea. To better understand the cellular mechanisms underlying RV‐diarrhea, the time‐course of RV replication, diarrhea and intestinal structural and enzyme responses were assessed in the preclinical piglet model. Formula‐fed, colostrum‐deprived piglets were infected on d4 postpartum with group A porcine RV strain OSU (P9[7],G5). Piglets were euthanized at 0, 4, 8, and 12 h, and 1, 2, 3, 4, 6 and 8 d post‐infection (PI). RV non‐structural protein 4 mRNA expression, a marker of RV replication, increased (p<0.001) at 2d and 4d PI, but returned to baseline by 6d in the jejunum. Diarrhea occurred at 2d, peaked at 3d, began to decrease by 4d, but persisted at 8d PI compared to 0h (p<0.05). Weight gain was positive throughout the study, except a transient loss on (−0.23 kg) 2d PI. Villus height was reduced 50% from 2–8d PI (p<0.05), but crypt depth was unaffected by RV infection. Jejunal lactase activity mirrored villus height and was reduced 3‐fold from 2 to 8d PI (p<0.05), whereas sucrase activity increased 2‐fold over 0h at 2 to 8d PI (p<0.05). Thus, diarrhea persisted beyond RV replication, supporting impaired villus morphology and impaired lactose digestive capacity as the underlying causes for persistent diarrhea following RV infection. Funded by USDA AG‐05‐34505.
The effect of high glucose upon the human plasmalemmal serotonin transporter (SERT)
Authors: Gonçalves Pedro , Araújo João R. , Martel Fátima
Published: June 2021

The aim of this work was to investigate the effect of short‐ and long‐term high glucose exposure on the plasmalemmal serotonin transporter (SERT)‐mediated uptake of 3H‐serotonin (5HT) by Caco‐2 cells. We verified that short‐term exposure of Caco‐2 cells to diabetic‐like high apical glucose levels (30 mM for 2 h or 40 mM for 1 h) decreased the uptake of 3H‐5HT by 20–30%. On the other hand, long‐term (21–24 weeks) exposure of the cells to diabetic‐like high (25 mM) glucose caused a 30% increase in the uptake of 3H‐5HT. Under these conditions, the affinity of the transporter for 5HT and noradrenaline was not significantly changed, and the inhibitory potencies of fluoxetine and desipramine were not changed as well. So, it appears that a long exposure to high glucose levels did not change the affinity of SERT for substrates and inhibitors. In conclusion, high glucose levels modulate SERT activity. A short‐term exposure of the cells to a high concentration of glucose decreases the activity of the transporter, whereas a longer exposure of the cells to a high concentration of glucose increases the activity of SERT, without interfering with the affinity of this transporter for agonists and antagonists. Supported by FCT (PTDC/SAU‐FCF/67805/2006).
Ventricular epicardial conduction slowing in the pressure loaded isolated murine heart
Authors: Wright Adam Thomas , Omens Jeffrey H , McCulloch Andrew D
Published: June 2021

The mechanisms by which mechanical stimuli alter the cardiac action potential and its propagation (mechanoelectric feedback, MEF) remain poorly understood. Previously, we reported that epicardial conduction velocity (CV) slows during ventricular filling in the isolated rabbit heart in the presence of 2,3‐butanedione monoxime (BDM), an electromechanical decoupler. This has not been reported in other species. Here we use optical mapping of the left ventricular (LV) epicardial surface of isolated beating murine hearts loaded with the voltage sensitive dye, di‐4‐ANNEPS to assess changes in action potential propagation at LV end diastolic pressures of 0 and 30 mmHg. In preliminary studies in the absence of BDM, pressure load resulted in a reversible 10–20% decrease in CV in the directions of fastest (CVmax) and slowest (CVmin) propagation during epicardial pacing (CVmax and CVmin dropped from 480 ± 2 and 208 ± 16 mm/s in the unloaded state to 432 ± 26 and 165 ± 5 mm/s in the loaded state). This drop in CV results in a phenotype promoting reentrant arrhythmias in the loaded state. This experimental design provides a means of studying MEF under physiological conditions and will be used to assess the contributions of stretch activated channels and passive electrical tissue properties to this observed slowing of conduction. This research has been funded by the National Science Foundation and the National Institutes of Health.
THE INSULAR HUB OF CEREBRAL CORTICAL SENSORY AND MOTOR REGIONS
Authors: Johnson John Irwin , Morris John Andrew , Fobbs Archibald J.
Published: June 2021

Insular cortex has strong connections with sensory pathways from, and motor pathways to, visceral structures. It adjoins ventral striatum, the site of processes related to behavioral motivation, rewards, punishments, and addictions. In addition to viscerally related inputs and outputs, insular cortex receives projections from all major sensory pathways, and is the site of interactions among multiple sensory modalities. There is close proximity between the insular projections from a given modality and other regions of cortex receiving projections from this same modality. Insular motor regions are somatotopically continuous with other motor regions. Insular cortex is thus a hub, from which other sensory and motor regions radiate outward on the brain surface. This is most evident in a cerebral cortex which has been unfolded and flattened into a two‐dimensional representation. The hub position of insular cortex, relative to visceral and somatic sensory and motor areas, supports current ideas about the role of the insula in the perception of bodily self image, and related behavioral governance. Supported by NSF grants IBN 0131267, 0131826, and 0131028.
Families and Schools for Health: Preliminary Findings from a Rural Obesity Intervention
Authors: Harrist Amanda W. , Laura Hubbs‐Tait , Topham Glade L. , Kennedy Tay S. , Page Melanie C.
Published: June 2021

Objective was to test the impact of two interventions in a sample of 1st graders (20 schools) at risk for obesity. Sessions were 90‐min/wk over 12 weeks. One targeted Family Food & Lifestyle (FL) variables only and one added a Family Dynamics (FD) psycho‐educational component. FL treatment (n = 33 > 75% BMI in 6+ sessions) provided education about healthy eating/exercise and behavior modification for lifestyle. FD intervention (n = 24 > 75% BMI in 6+ sessions) also targeted psychosocial issues (e.g., parenting, communication). Mother report of food practices/concerns on CFQ were compared from baseline to post‐intervention, controlling for baseline differences. Analyses here were conducted for overweight children (BMI > 95%). Intervention effects were significant (p < .05, df (1, 14)) in half of analyses conducted and include: Concern about child dieting, FL>Control, F= 4.52; Need to guide or child eats too much junk food, FD>FL, F= 5.22; Careful that child eats enough, FD>Control, F= 6.15, FL>Control, F= 4.39; Tracking child's high‐fat food, FD>Control, F= 5.60, FL>Control, F= 17.79; Tracking fruit/vegetables, FD>Control, F= 9.09, FD>FL, F= 10.08; Tracking dairy, FD>Control, F= 4.48; and Tracking nutritious foods child eats, FL>Control, F= 4.85. Analysis for children > 85% and > 75% BMI yielded similar results but more effects were evident among those > 95%. Results suggest impact of both treatments. USDA Grant 05545
Effect of running and swimming exercise training on skeletal growth and bone microstructure of the American alligator (Alligator mississippiensis) with and without the cardiac shunt
Authors: Owerkowicz Tomasz , Tsai Henry P. , Sanchez Linda D. , Gwalthney June C. , Eme John , Blank Jason M. , Hicks James W.
Published: June 2021

Exercise affects the vertebrate skeleton via biomechanical and biochemical stress. Exercise‐induced strains can elicit increased primary bone formation and/or secondary remodelling of limb bones. Exhaustive exercise results in high lactic acid loads, which may require buffering by bone mineral and thus influence bone remodelling. Cardiac shunting in crocodylians is thought to divert protons to the enteric circulation and reduce acidity of systemic arterial blood in exercise‐stressed animals. We subjected juvenile alligators to exhaustive exercise (running or swimming) every other day for 17 months, and compared their skeletons with those of sedentary controls. Prior to exercise training, half the animals in each group had their left aorta surgically occluded, thus rendering them incapable of cardiac shunting. Alligators were fed twice weekly, and given injections of fluorescent dyes to determine bone deposition rates. Preliminary data show that bone formation rate at the femoral midshaft correlates with whole‐body growth rate in all groups, but is significantly higher in treadmill runners. This suggests that terrestrial, but not aquatic, exercise stimulates bone growth. Loss of cardiac shunting did not affect bone formation or remodelling, which suggests cardiac shunting does not protect skeletal integrity in alligators. Funded by NSF IOB 04445680 to JWH. TO and JMB supported by NIH 2T32AR047752.
Wnt signaling and the evolution of pattern formation
Authors: Wikramanayake Athula H , Kumburegama Shalika , Peng Jeff , Wijesena Naveen
Published: June 2021

Evolution of gastrulation was crucial for metazoan diversification, but the molecular origins of developmental polarity that led to the onset of gastrulation mechanisms are unknown. Nuclear ß‐catenin marks the site of gastrulation in cnidarians and several bilaterians, raising the possibility that evolution of mechanisms to asymmetrically activate Wnt signaling in embryos may have been causal in evolution of gastrulation. We have shown that localized activation of Dishevelled (Dsh), a key regulator of Wnt/ß ‐catenin signaling, plays a crucial role in germ layer segregation and gastrulation in sea urchins and in the cnidarian Nematostella vectensis. Dsh also regulates the Wnt/Planar Cell Polarity (PCP) pathway, which mediates vertebrate gastrulation movements. In Nematostella, blocking Dsh signaling in the PCP pathway, or blocking Strabismus function using morpholinos blocks gastrulation, suggesting that Wnt/PCP signaling mediated gastrulation movements in the last common ancestor to cnidarians and bilaterians. Because Dsh can regulate multiple Wnt pathways, it may have been a critical link between evolution of cell fate specification and evolution of gastrulation morphogenesis. Our results also support the hypothesis that one ancient function of PCP signaling was to mediate the developmental asymmetry that led to the evolution of gastrulation. (Supported by NSF grants IOS 0720365 and IOS 0754323)
Embryonic derivation and segmentation of the bony skull are not conserved among vertebrates: data from amphibians
Authors: Hanken James , Gross Joshua B.
Published: June 2021

Fundamental aspects of cranial anatomy in vertebrates were resolved initially well before the delineation and broad acceptance of Darwinian evolution, let alone modern studies of molecular genetics and cell‐lineage analysis and broad recognition of the prominent role of the embryonic neural crest. Contemporary work has validated many classical observations, yet others mandate reassessment and reevaluation of several underlying assumptions and conventions. Head segmentation is perhaps the most classical topic of all. Compelling accounts regarding the nature and extent of head segmentation abound, but the empirical base on which such accounts are grounded is surprisingly thin. We have utilized a novel transgenic line of the clawed frog, Xenopus laevis, to map neural‐crest derivation of the bony adult skull. This method circumvents technical limitations that have hindered previous studies of the embryonic origin of adult‐specific features in frogs, which form first at metamorphosis. The bony skull of Xenopus is segmented, but in important ways that differ from both those predicted by conventional theories of cranial organization and those reported for other vertebrates. Patterns of cranial segmentation are evolutionarily labile and susceptible to modification associated with the origin of specialized life‐history and developmental modes. Supported by NSF (EF‐0334846; AmphibiaTree).
Macrophage Killing of Intracelluar Pathogen Fransicella tularensis: Functional Genomic Analysis
Authors: Zhou Hongwei , Browning Erica , Tan Fengxiao , Kobzik Lester
Published: June 2021

Francisella tularensis (FT) is a highly virulent, facultative intracellular bacterium which causes tularemia, and is a potential bioweapon. Interferon‐γ has been reported to play a crucial role in inhibition of intracellular replication of FT in human monocyte‐derived macrophages and in mice, but the mechanisms for this protective effect are poorly characterized. Our hypotheses were that IFN‐g limits intracellular pathogen FT by activation of specific genes, and functional genomic screening can identify these genes. We transduced human macrophage THP‐1 cells with a genome‐wide lentiviral siRNA library targeting ~47,000 human genes and ESTs. We screened for RNA interference resulting in a phenotype that failed to show the beneficial effects of IFN‐g on intracellular FT. We sorted macrophages with highest GFP‐FT load. The siRNA inserts were amplified by PCR from genomic DNA in these cells, and siRNA targets were identified by cloning and DNA sequencing. From 5 sorting experiments, a total 4078 clones were sequenced, and 3386 genes/transcripts/ESTs were identified. We found 212 genes of interest, with 61 found in > 3 sorts, and 2 found in 4 sorts. Validation is underway using PCR arrays to confirm expression in IFN‐g‐activated primary human macrophages challenged with FT. This approach may prove useful to identify mechanisms in macrophage innate immunity against intracellular pathogens.

Supported by DTRA grant (Defense Threat Reduction Agency)
Maternal immune stimulation influences behavior associated with congenital toxoplasmosis
Authors: Hrubec Terry C , Goodwin David , Lindsay David S , Holladay Steven D
Published: June 2021

The protozoan parasite, Toxoplasma gondii, can have devastating effects on the developing fetus if infection occurs during pregnancy. Congenital toxoplasmosis results in stillbirths, hydrocephalus, blindness, mental retardation, and neurological and behavioral disorders. There is mounting evidence for a link between toxoplasmosis and mental syndromes such as schizophrenia, bipolar disorder, major depression and possibly autism. In mice, stimulating the maternal immune system during the periconceptual period dramatically reduces teratogen induced birth defects. Previously we demonstrated that maternal immune stimulation prevents fetal mortality from congenital toxoplasmosis. We wished to see if we could prevent toxoplasma induced behavioral changes as well. Pregnant mice were immune stimulated with IFNγ and were then infected with T. gondii tissue cysts. Congenitally infected and control pups were assessed for 31 standard FOB neurobehavioral assessments at 5 and 8 weeks of age. Significant differences in activity level, visual scores and click responses between treatment and gender were observed. These results may lead to development of a treatment or prevention for congenital toxoplasmosis and associated neurobehavioral disorders. Supported by USDA AD‐421 137196.
Alternate protein splicing mechanisms: A directed evolution approach
Authors: York Daniel J. , Nadelson Adam C. , Reitter Julie N. , Mills Kenneth V.
Published: June 2021

Inteins are intervening polypeptides that are able to direct their excision from the flanking polypeptides, called exteins, as well as facilitate the ligation of the exteins. This process, called protein splicing, produces the functional protein. We want to determine if inteins that splice by the standard mechanism also can splice via alternate mechanisms. First, we have inactivated splicing of the RecA intein from M. tuberculosis by mutating the N‐terminal Cys to Ala. Using saturation mutagenesis and error prone PCR, we generated random mutants that we subjected to a genetic screen to select for complementary mutations that may allow the modified intein to facilitate splicing by bypassing the first step. To select for complementary mutants, we used E. coli harboring a plasmid with the kanamycin resistance gene, aminoglycoside phosphotransferase, interrupted by the RecA intein, such that bacterial growth should depend on efficient splicing. We have been able to mutate the RecA intein and select inteins that allow for growth on kanamycin. However, these inteins do not promote splicing in our biochemical assay, which suggests that the intein may allow for a functional aminoglycoside phosphotransferase in the absence of splicing. This material is based upon work supported by the National Science Foundation under Grant No. 0320824 and CAREER grant No. 0447647.
Both Subunits of Thermotoga maritima ADPGlucose Pyrophosphorylase are Required for Optimal Acitvity and Allosteric Regulation
Authors: Matsui Mikiko , Lim Shannon , Harake Tala , Guandique Erick , Tran Margaret , Reyes David , Orry Andrew , Meyer Christopher R
Published: June 2021

The T. maritima genome harbors two ADP‐glucose pyrophosphorylases (ADPG PPases), glgC and glgD, which share only 36.9% identity. Activity assays revealed negligible glgD activity alone in accord with molecular modeling predictions. However, glgD showed 20‐fold stimulation when combined with glgC. The kinetic parameters for glgC alone and the glgD/C complex were determined. The glgC protein was insensitive to a screen of 15 known ADPG PPase effector molecules, as predicted from modeling, but glgD/C exhibited 2.2‐fold activation by FBP. The following mutations were introduced to restore glgD activity: K12R, S13G, D14S, S25K, A26P, F189S, N240D. This altered protein was inactive alone but in complex with glgC displayed a 2.7‐fold increase over wild‐type glgD/C, indicating that the pyramided mutations effectively resurrected glgD activity in the presence of glgC. The glgD (R23A,D344N) and glgC (I24R,A347D,E348R,N349G) proteins were generated based on the molecular modeling of a hypothetical salt bridge between the N and C‐terminal domain involved in allostery. The glgD AN protein was not as responsive to FBP and the glgC RDRG enzyme exhibited partial FBP activation. Supported in part by NSF Grant 0448676.
IMAGEtags for imaging gene expression in living cells in real‐time
Authors: Ilgu Muslum , Shin Ilchung , Haarberg Hans Eirik , Gupta Vinayak , Kraus George A , Nilsen‐Hamilton Marit
Published: June 2021

The use of green fluorescent protein (GFP) and luciferase in cellular imaging has enabled significant advances in cellular imaging in vitro and in vivo. However, these imaging systems have the disadvantages of very slow response times and a high energy costs for synthesizing and maintaining reporter protein levels. For example, they are not amendable to imaging changes in gene expression in real time. Also, the energy requirements of protein reporter systems are potentially debilitating in vivo when cells such as stem cells need to proliferate and populate a niche in a damaged tissue where the oxygen and nutrient concentrations are limiting. Therefore, our goal is to establish an alternative reporter system that can be used for real‐time imaging and for imaging stem cells in vivo with minimal metabolic perturbation. This new reporter system involves the use of IMAGEtags (Intracellular Multiaptamer Genetic tags), which are strings of RNA aptamers expressed from a promoter of choice. To establish this RNA‐based reporter system, we utilized available aptamers that recognize small target molecules such as neomycin, tobramycin and theophylline and constructed vectors that express multiple repeats of these aptamers. The target molecules were labeled with fluorescent moieties for intracellular imaging. To increase signal/noise the aptamer targets were separately labeled with fluorescent FRET pairs. IMAGEtags are currently being developed for reporting gene expression changes in yeast and cultured mammalian cells. Once optimized for imaging, this new reporter system will have imaging applications such as real‐time detection of changes in gene expression and mRNA processing, and tracking cell movements in vivo.

Supported by grants from the NIH (R01HL078659) and the DOE (OBER).
Evaluation of His‐tag and immunoprecipitation procedures for recombinant protein purification
Authors: Lin Rui , Cao Heping
Published: June 2021

Histidine (His)‐tag is widely used for affinity purification of recombinant proteins, but the yield and purity of expressed proteins are quite different. Little information is available about quantitative evaluation of this procedure. The objective of current study was to evaluate His‐tag procedure quantitatively and to compare it with immunoprecipitation using radiolabeled tristetraprolin (TTP), a zinc finger protein with anti‐inflammatory property. Human embryonic kidney 293 cells were transfected with wild‐type and nine mutant plasmids with single or multiple phosphorylation site mutation(s) in His‐TTP. These proteins were expressed and mainly localized in the cytosol of transfected cells by immunocytochemistry and confocal microscopy. His‐TTP proteins were purified by Ni‐NTA beads with imidazole elution or precipitated by TTP antibodies from transfected cells after being labeled with [32P]‐orthophosphate. The results showed that 1) His‐tag purification was more effective than immunoprecipitation for TTP purification; 2) mutations in TTP increased the yield of His‐TTP by both purification procedures; and 3) mutations in TTP increased the binding affinity of mutant proteins for Ni‐NTA beads. These findings suggest that bioengineering phosphorylation sites in proteins can increase the production of recombinant proteins (Supported by NIH‐NIEHS and USDA‐ARS Intramural Research Programs).
Regulation of High Mobility Group A1 expression by the Adenomatous polyposis coli (Apc) tumor suppressor gene
Authors: Bush Bethany Marie , Hurlbert Jason , Sumter Takita Felder
Published: June 2021

High Mobility Group A1 (HMGA1) is disregulated in neoplasias and is a prominent biomarker for colon cancer. However, the molecular networks involved in HMGA1‐mediated transformation are only partially understood. Mutations in the adenomatus polyposis coli (Apc) gene are associated with the earliest stages of colon carcinogenesis. These mutations activate the Wnt signaling pathway, resulting in stabilization of beta‐catenin which then binds to T‐cell factor‐4 (Tcf‐4) causing subsequent increases in target gene expression. We explored the role of the Wnt pathway in HMGA1 regulation. In Apc(Min/ +) mice expressing truncated Apc, we observed 9‐fold greater HMGA1 mRNA levels relative to mice bearing wild‐type Apc. HMGA1 protein levels were also significantly elevated in Apc (Min/+) intestinal tumors. To assess the effects of wild type Apc recovery, we used HT‐29 cells (human colorectal carcinoma cell line with truncated Apc) in which wild type Apc has been introduced under the control of a zinc inducible promoter. Induction of full length Apc caused down‐regulation of HMGA1 at the translational level suggesting a novel role for HMGA1 in Wnt signaling. Current studies are aimed at understanding the mechanism by which HMGA1 expression is regulated using MALDI‐TOF mass spectrometry. Our data implicates Wnt signal transduction as a regulator of HMGA1 in colon cancer. Work funded by NIH P20RR16461, NSF 0542242; 0722722.
3,4‐Disubstituted benzamides protect CD4+ T‐cells from acute apoptotic stimuli
Authors: Nguyen Jeffrey , Walton Andrew , Ellenberger Tom , McDunn Jonathan
Published: June 2021

Sepsis is a pathologic condition with extensive lymphocyte apoptosis. We screened 32,000 compounds and identified 15 that prevent CD4+ T‐cell apoptosis. Most hits also augmented Th1 cytokine production. A 3,4‐disubstituted benzamide was selected for structure‐activity studies. Derivatives were synthesized in four steps, installing different functional groups at the 1 and 4 positions. Compounds were evaluated in apoptosis, cytokine secretion and protein phosphorylation assays. Apoptosis was induced by lyophilized E. coli and caspase‐3 activity was measured. Cytokine secretion was stimulated using agonistic antibodies to CD3 and CD28. Protein phosphorylation was determined after 15 minute incubation with compound. The average yield of 3,4‐disubstituted benzamides was 43%. The first generation library yielded 2 compounds that: protected cells from apoptosis, promoted Th1 cytokine release, and led to ~30% increase in pAkt and ~21% increase in pIκBα without altering MAP kinase phosphorylation. These compounds modulate CD4+ T‐cell activity by removing the need for a costimulatory signal and may represent a new approach to the treatment of sepsis. Supported by DTRA and WUSM.
Genetic and environmental influences on eating behavior – a study of twins reared apart
Authors: Elder Sonya J. , Neale Michael C. , Fuss Paul J. , McCrory Megan A. , Bouchard Thomas J. , Saltzman Edward , Roberts Susan B.
Published: June 2021

Little information exists on the influence of genetic versus environmental factors on eating behavior. As part of the Tufts Twin Study, we assessed genetic (G), common environmental (C) and unique environmental (E) influences on eating behavior constructs and subscales by using the Eating Inventory (EI) in monozygotic twins reared apart (29 pairs) or together (49 pairs) (mean of all subjects±SD: age 36.2±13.6, BMI 24.5±4.5), and determined the extent to which G, C and E influences are shared by restraint (RES), disinhibition (DIS), and hunger (HUN) using model‐fitting analyses. The variance of most EI variables was primarily explained by E, which most strongly influenced external locus for HUN and strategic dieting behavior of RES (explaining 71% and 69% of variance, respectively). However, G had a significant influence on RES, internal locus for HUN, and emotional and situational susceptibility to DIS (with heritabilities of 52%, 50%, 55% and 38%, respectively). Multivariate analyses showed that DIS and HUN share a common influence, while RES appears to be a distinct construct. These results provide insight into the etiology of eating behavior by identifying specific constructs of eating behavior that are heritable and others that are primarily influenced by E. Funding: NIH grants R01 DK046124, 5T32 HL069772, MH‐65322, DA‐18673, and USDA grant 58‐1950‐4‐401.
Diet quality is related to eating competence in low‐income females in Pennsylvania
Authors: Bailey Regan Lucas , Stotts Jodi L , Wall Denise E , Lohse Barbara A
Published: June 2021

Eating competence (EC) denotes an intrapersonal approach to eating behavior and has been positively associated with diet quality and health. Little is known about the relationship between EC and dietary patterns (DP) among low‐income (LI) females. EC and DP were examined in LI females (n=149, age range 18‐60) in Pennsylvania. EC was measured by the ecSatter Inventory (ecSI); 44.3% (n=66) were defined as eating competent (mean ecSI score was 29.54±7.7). Three, 24‐hour dietary recalls were collected. A principal components analysis with varimax rotation revealed two DPs. DP1 consisted of refined grains, added fats, sweetened beverages, and whole fat dairy; DP2 was characterized by fruits, vegetables, and whole grains. DP1 was also associated with lower intakes of most micronutrients and fiber; DP2 related to higher EC, Healthy Eating Index and micronutrients and lower percent energy from fat. EC was associated with higher intakes of fiber, vitamin C, most B vitamins, vitamin A and many minerals including iron. Recommendations for 5 or more nutrients were not met by 79.5% of non‐eating competent females and by 56.1% of eating competent females. Mean number of nutrients with an at‐risk intake level was lower for eating competent participants (P=.02). This association provided a rationale for focusing nutrition education on EC as a means to improve the diet quality of LI individuals. Funding provided by USDA FNS SNAP‐ Ed.
A comparison of nutrient density scores for orange vegetables: A call for inter‐ and intra‐group variety when recommending vegetable intake
Authors: ZANOVEC MICHAEL , O'Neil Carol E. , Nicklas Theresa A.
Published: June 2021

The objective of this study was to compare inter‐ and intra‐group nutrient density (ND) scores of orange vegetables based on type and preparation. ND scores of carrots, sweet potatoes, pumpkin, and 3 varieties of winter squash were calculated using 6 methods. Nutrient profiles/100 g of food were calculated for each vegetable for different preparations. Nutrient analyses were from the Nutrition Data System. ND scores were calculated using: 1) Naturally Nutrient Rich, 2) Calorie for nutrient, 3) Ratio of Recommended to Restricted nutrient, 4) Fulgoni nutrient for calorie, and two ad hoc nutrient for calorie (5 & 6) methods. Inter‐group comparisons between vegetables were based on the mean ND scores for each of the six methods, whereas intra‐group differences were based on the ND scores by preparation. Sweet potato ranked highest with 1, 5, and 6. Carrots ranked highest with 2 and 4; pumpkin ranked highest with 3. When comparing the ND scores of each vegetable by preparation, cooked carrots ranked consistently higher when compared with raw and canned. Baked sweet potatoes ranked highest and canned lowest. Butternut squash ranked highest among the squashes for all 6 methods used. Canned pumpkin ranked higher than cooked. Results illustrate the complexity of calculating ND scores and highlight the need to consider inter‐ and intra‐group differences when recommending vegetable intake. Supported by USDA.
Morning nutrition and executive function processes in preadolescents: gender variations in phasic modulation of frontal EEG theta activity during a go/ no‐go task
Authors: Pivik R. T. , Dykman R. A.
Published: June 2021

Frontal EEG theta activity has been related to executive functions (i.e., goal‐directed behavior such as inhibition and flexibility of action). We studied the effects of morning nutritional status on frontal theta‐executive function relationships using stimulus‐locked responses [event‐related increases (ERS) and decreases (ERD) in synchronization] of 4‐6 Hz activity in a cued visual go/no‐go task in healthy, right‐handed children (8‐11 yrs. old; IQ > 80). Children were tested after overnight fasting (Phase 1), and again (Phase 2) after eating a standardized breakfast (n = 30; 15 males) or while continuing to fast (n = 30; 15 males). Performance measures‐accuracy and reaction time (RT)‐were obtained. Data were analyzed using ANOVA procedures with post‐hoc t‐tests. Response accuracy was similar across groups. Relative to Phase 1, RT slowed in Phase 2 for both groups, but the decrease was only significant for fasting females (p < .01). Consistent with increased executive function demands associated with response inhibition, ERS was greater to no‐go than go stimuli (p < .05) for both groups. Nutritional status did not significantly influence ERS responses in females, but Phase 2 responses to no‐go stimuli were decreased in fasting, but not fed, males (p < .05). These data suggest a greater sensitivity of frontal executive function processes to morning nutritional status in preadolescent males than females. (Supported by USDA ARS CRIS 6251‐51000‐002‐03S)
Muscle atrophy and long bone osteopenia are attenuated with n‐3 PUFA in a mouse model of hindlimb suspension
Authors: Watkins Bruce A , Hutchins Heather L , Torregrosa‐Allen Sandra E , Hannon Kevin M , Li Yong
Published: June 2021

Muscle atrophy and osteopenia are common issues associated with aging and disease. This study was conducted to evaluate the effects of long chain n‐3 PUFA [eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids] on muscle and bone loss in a mouse model of induced hindlimb atrophy. Mouse hindlimb suspension (HS) is used as a disuse model to induce muscle atrophy and bone loss compared to weight bearing (WB). Six‐wk‐old male mice (ND4 Swiss Webster) were fed a control (n‐6 PUFA, n= 26), a moderate n‐3 PUFA (EPA + DHA 12 g/kg, n = 20), or a high n‐3 PUFA (19.4 g/kg, n= 21) diet for 2 wks. After pre‐feeding, mice were subjected to a baseline DEXA scan and then assigned to the WB or HS groups. Upon 14 days of HS or continued WB, mice were anesthetized for a repeated DEXA scan after which the mice were euthanized, and tissues were collected for analyses. Mice fed n‐3 PUFA demonstrated an elevation of EPA and DHA and reduced arachidonate in serum and tissues. HS resulted in fat, muscle mass and bone mineral content (BMC) losses. Bone ash was conserved in the HS n‐3 PUFA fed groups compared to control HS mice, which supports the protection of BMC/body wt also found in the n‐3 fed groups. There was no additional protection of bone in the high n‐3 diet group compared to the moderate group. Based on this study in mice and other investigations with rodents, the n‐3 PUFA appear to attenuate the physiological changes that occur with HS. Supported by USDA funds.
K12‐biotinylated histone H4 is enriched in human telomeric repeats
Authors: Wijeratne Subhashinee Samudra Kumari , Zempleni Janos
Published: June 2021

Covalent modifications of histones play a role in regulating telomere attrition and cellular senescence. Biotinylation of lysine (K) residues in histones, mediated by holocarboxylase synthetase (HCS), is a novel diet‐dependent mechanism to regulate chromatin structure and gene expression. We have previously shown that biotinylation of K12 in histone H4 (H4K12bio) is a marker for heterochromatin and is enriched in pericentromeric alpha satellite repeats. Here we hypothesized that H4K12bio is also enriched in telomere repeats. We used human IMR‐90 lung fibroblasts and immortalized IMR‐90 cells overexpressing human telomerase (hTERT) to quantify H4K12bio in telomeres from young and old cells. About 30% of the histone H4 molecules in telomeres were biotinylated in position K12. The abundance of H4K12bio in telomeres decreased by 42% during telomere attrition in senescent IMR‐90 cells; overexpression of telomerase prevented the loss of H4K12bio. Possible confounders such as decreased expression of HCS and biotin transporters were formally excluded. These preliminary data suggest that H4K12bio might play a role in telomere structures. (Supported by UNL ARD Hatch Act funds, NIH DK063945, DK077816 and ES015206, USDA CSREES 2006‐35200‐17138, and NSF EPSCoR EPS‐0701892.)
Green tea extract (GTE) reduces hepatic injury by inhibiting lipid accumulation and peroxidation in steatotic livers of obese mice
Authors: Chung Min‐Yu , O'Connor Meeghan , Park Hea Jin , DiNatale Dana , Koo Sung I. , Manautou Jose E , Bruno Richard S
Published: June 2021

Steatotic livers are vulnerable to oxidative insults that lead to hepatic lipid peroxidation and injury. Based on our finding that green tea extract (GTE) protected against hepatic steatosis and injury, we hypothesized that GTE would decrease hepatic lipid peroxidation and inflammation. Obese (ob/ob) mice (n = 38) were fed a diet containing GTE at 0, 0.5, or 1%, and lean mice (n = 12) were fed 0% GTE for 6‐wk. Then, hepatic lipid and malondialdehyde (MDA), serum alanine aminotransferase (ALT), and inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF)‐α were measured. Body weight was 33% greater (p<0.05) in obese compared to lean controls and was 18% lower in obese mice fed GTE at 1%. Hepatic lipid was 2.3‐times higher in obese mice and GTE at 1% reduced it by 13%. Hepatic MDA was 40% higher in obese compared to lean mice and GTE reduced it 16‐22%. Serum ALT was 7.3‐fold higher in obese controls and GTE at 1% decreased it 25%. MDA was correlated (p < 0.05) to ALT (r = 0.358) and total lipid (r = 0.383). Immunohistochemical evidence suggested that obese mice had greater hepatic iNOS expression and may be decreased with GTE at 1%. TNF‐α expression was unaffected by GTE. These data suggest that GTE protects against hepatic injury by decreasing hepatic lipid accumulation, peroxidation, and potentially obesity‐induced inflammation. Supported by a USDA‐NRI grant (2007‐02303) to RSB.

Grant Funding Source

USDA
Cognitive status and vitamin K status in older men and women
Authors: Truong Jennifer T , Scott Tammy , Yoshida Makiko , Shea M. Kyla , Booth Sarah L
Published: June 2021

Vitamin K has been implicated in the regulation of sphingolipid metabolism. In rats, vitamin K deficiency decreases sphingolipid concentrations, resulting in changes in behavior, but not short‐term memory. The association between vitamin K intake and cognition in humans is not well studied. In 379 men and women (60‐80 y, 59% F) participating in a randomized controlled trial to assess the effect of 3‐year vitamin K supplementation (500 µg phylloquinone/day) on bone mineral density, we compared cognitive function by Trail Making Tests A and B and the Digit Symbol‐Coding Test (DSCT) between those receiving vitamin K vs. control. There were no significant differences in medication use known to alter cognitive function (i.e. statins and psychotropic medications) between groups. There was a non‐significant trend for a group*baseline vitamin K status interaction (p=0.054) (= 0.5 mmol/L vs. > 0.5 mmol/L) for results on the DSCT, so stratified analyses were conducted. There was no significant difference in all three cognitive function tests between the control and the vitamin K group, even after adjustment for age, sex, education, physical activity and alcohol intake. In this cohort of older, relatively healthy adults, vitamin K supplementation in amounts achieved in diet did not confer additional benefits on cognitive function. Research support: NIA A914759 & USDA ARS (cooperative agreement 58‐1950‐7‐707)
Factors associated with academic success in the sciences
Authors: McGonagle Alyssa , Winston Jeffrey , Barnes‐Farrell Janet , Freake Hedley C
Published: June 2021

This study investigated social and psychological factors relevant to students' academic performance in the sciences, as part of an evaluation of STRONG‐CT, a program striving to increase representation of minority and first‐generation students in the sciences. College students from a large university in the northeastern U.S. volunteered to complete an online survey of social and psychological factors that were theorized to relate to academic performance in the sciences; in addition, science course grades and SAT scores were obtained from a university database for each participant. Hierarchical multiple regression was used to analyze the data; Math SAT scores were entered in the first step to control for their impact on grades and all study variables were entered in the second step. Levels of self‐efficacy for tasks vital to learning were positively related to science GPA; levels of college‐related stress were negatively related to science GPA. Additionally, students who reported going to professors for extra help were found to have higher science GPAs than those who did not. STRONG‐CT students demonstrated higher levels of self‐efficacy, were more likely to meet with professors, and reported studying for more hours per week than other students. STRONG‐CT students also had higher science GPAs than a control group. Supported by NSF‐0525379.
Does Yck1/2 phosphorylation affect septin‐dependent compartmentalization in yeast cells?
Authors: Saulsbury Spencer , Brame Cynthia J. , Robinson Lucy C.
Published: June 2021

Yeast casein kinase 1 (CK1) proteins Yck1 and 2 are redundant and essential plasma membrane‐associated protein kinases in budding yeast that are required for cellular morphogenesis and cytokinesis. We identified Ser548 in Shs1 as a potential Yck1/2 phosphorylation site using a comparative phosphoproteomic approach. Shs1 is one of five septin proteins that are essential for several aspects of cell division, including mother‐bud membrane compartmentalization. Septin activity is regulated by phosphorylation, and septin ring formation does not occur properly in cells expressing the low activity yck2ts mutant protein. We generated cells expressing mutant Shs1 proteins altered at the putative Yck2 phosphorylation site and tested the ability of these cells to maintain membrane compartmentalization. Cells expressing S548A or S548E Shs1 maintained appropriate endoplasmic reticulum membrane separation during bud formation, as assessed by low fluorescence recovery after photobleaching of GFP‐tagged Sec61. Similar results were observed in cells expressing the yckts protein. We conclude that phosphorylation of S548 on Shs1 is not required for this function of the septin ring. Supported by NSF grant MCB‐0517204.
Inhibition of NOD2 Signaling and Target Gene Expression by Curcumin
Authors: Huang Shurong , Zhao Ling , Kim Kihoon , Lee Dong Seok , Hwang Daniel
Published: June 2021

NOD2 is an intracellular pattern recognition receptor that recognizes a conserved moiety of bacterial peptidoglycan and subsequently activates proinflammatory signaling pathways, including NFκB pathways. Mutations in NOD2 have been implicated in the development of inflammatory granulomatous disorders, such as Crohn's disease and Blau syndrome. Many phytochemicals possess anti‐inflammatory properties. We carried out a series of molecular and biochemical studies to determine whether any of these phytochemicals might modulate NOD2‐mediated immune responses. We have demonstrated that curcumin, a polyphenol found in the plant Curcuma longa, and parthenolide, a sesquiterpene lactone, suppress both ligand‐induced and lauric acid‐induced NOD2 signaling, leading to the suppression of NFκB activation and target gene IL‐8 expression. The suppression is mediated through the inhibition of NOD2 oligomerization and subsequent inhibition of the phosphorylation of IκBα and p65 proteins. These results show that curcumin and parthenolide can directly inhibit NOD‐mediated signaling pathways at the receptor level and suggest that the activation of NOD2 and subsequent immune and inflammatory responses can be inhibited by these phytochemicals. Research support: NIH grants DK064007, DK41868, and CA75613, USDA grant 2001‐35200‐10721, and American Institutes for Cancer Research grant 01A095Rev.