bluuebunny/crossref_metadata_2025 huggingface dataset:
Macrophage migration inhibitory factor is required in concanavalin A‐induced autophagic cell death of human hepatoma (151.9)
Authors: Lai Yen‐Chung , Yeh Trai Ming , Chuang Yung‐Chun
Published: June 2021

Concanavalin A (Con A) is a lectin, which can induce autophagic cell death of hepatoma cells and is considered as an anti‐hepatoma agent. Macrophage migration inhibitory factor (MIF) is a pro‐inflammatory cytokine, which can induce autophagy of hepatoma cells. However, it is unclear whether MIF is required in Con A‐induced hepatoma cell death. In this study, we demonstrated that MIF expression and secretion were increased in Con A‐stimulated human hepatoma cell lines, HepG2 and HuH7. In addition, Con A‐induced autophagy of hepatoma cells was blocked in the presence of MIF inhibitor (ISO‐1). Knockdown of endougenous MIF by small hairpin RNA (shRNA) further confirmed MIF is required in Con A‐induced autophagy of hepatoma cells. These results may explain why lack of MIF can protect mice against Con A‐induced liver injury and suggest the combination of Con A with MIF may represent a better therapy approach against hepatic cancer.
Exogenous expression of the Entamoeba histolytica adhesion protein EhADH112 in MDCK cells facilitates parasite invasion by altering tight junctions (152.1)
Authors: Betanzos Abigail , Cuellar Patricia , Azuara‐Liceaga Elisa , Bañuelos Cecilia , González‐Mariscal Lorenza , Schnoor Michael , Orozco Esther
Published: June 2021

Entamoeba histolytica is the causative agent of human amoebiasis. Infection is initiated by interaction of the pathogen with intestinal epithelial cells. This interaction leads to disruption of intercellular structures such as tight junctions (TJ). We recently showed that the E. histolytica complex EhCPADH112, formed by the EhADH112 adhesin and the EhCP112 cysteine protease, participates during epithelial invasion. To elucidate the particular role of EhADH112 in this process, we overexpressed this adhesion molecule in MDCK epithelial cells. Expression of EhADH112 was proven by RT‐PCR and western blot assays. EhADH112 localized at the plasma membrane of epithelial cells, similar to its location in trophozoites. EhADH112 improved adhesion between epithelial cells induced by an increase in TER and expression of TJ proteins. Importantly, infection of MDCK cells expressing EhADH112 with trophozoites caused a faster drop in TER and more epithelial damage compared to control cells; suggesting that EHADH112 in epithelial cells serves as a ligand for the parasite, facilitating trophozoite entrance into the epithelium. Combined with our previous results that EhADH112 localizes to TJ after secretion by trophozoites, EhADH112 may be a promising new target to prevent amebiasis.

Grant Funding Source: Supported by Institute of Science and Technology of the Federal District (64/2012)
Cell and molecular events comprising collateral vessel development and remodeling (19.1)
Authors: Tomanek Robert
Published: June 2021

Although the relevance of coronary collaterals was debated for many years, more recent studies utilizing more acurate methods for measuring collateral flow have shown that collateral flow can minimize infarct size. The development of coronary collateral vessels by pressure gradients (fluid shear stress), involving the recruitment of multiple genes, including VEGF and FGF, is well documented. However, there is also more recent evidence that ischemia, independent of elevated shear stress can also stimulate collateral growth under certain conditions. Collateral growth is a form of arteriogenesis that involves: 1) endothelial deformation, 2) increases in NOS and NO, 3) VEGF activation of MCP‐1, 4) monocyte and T‐cell recruitment and migration and 5) remodeling of the vessel. The remodelinig process requires proliferation of endothelial cells and vascular smooth muscle, which results in an increase in vessel diamter. Important stimuli for collateral growth include brief, repeated, as well as gradual coronary artery occlusions.
Genetic determinants of the native collateral circulation (19.3)
Authors: Faber James , Zhang Hua , Lucitti Jennifer , Sealock Robert , Moore Scott
Published: June 2021

Native (pre‐existing) collaterals connect adjacent arterial trees and limit ischemic tissue injury. I will review recent findings that collaterals have distinct features, including a unique process of formation and specialized endothelial and smooth muscle cell phenotypes. Moreover, the number and diameter of collaterals (“extent”) in tissues varies widely among different mouse strains and healthy humans, associated with wide differences in the severity of ischemic damage. A single locus, Dce1, is responsible for ~80% of the variation in mouse. Ongoing work is seeking to identify the underlying gene(s) and pathway, and whether this locus links to variation in collateral extent in humans. If so, Dce1 and its up‐ and down‐stream partners could provide markers for collateral abundance to help stratify patients for treatment options, to assess risk‐severity before disease occurs, and to provide targets for therapies to stimulate new collateral formation. Recent studies in mice have found that such “neo‐collateral” formation can be induced to occur rapidly after acute MI, resulting in increased collateral‐dependent flow and reduced infarct volume. Besides genetic factors, “environmental “ factors, ie, aging and other cardiovascular risk factors, cause reduced collateral extent, resulting in more severe ischemic injury. Studies are under way to identify the underlying mechanism and strategies to prevent it.
What makes connections effective? An introduction to model‐based measures of brain connectivity (21.1)
Authors: Gitelman Darren
Published: June 2021

Human brain mapping has traditionally concentrated on the localization of functions, with the primary question being where in the brain, activity for a particular function can be mapped to or located. This focus on localization or functional segregation has been critical for demonstrating the correspondence between specific brain regions and functions such as the perception of color and visual motion among others. However, it has also been recognized that complex behaviors defy an easy one‐to‐one correspondence between functions and single brain regions; instead these behaviors emerge from the integrated activity of multiple cortical and subcortical areas or brain networks. Brain network connectivity can be examined on several levels. Structural connectivity defines the general organization of the brain, but it does not tell us, which connections are engaged for a particular brain function. Functional connectivity highlights regions with similar time courses of activity. However, since functional connectivity is based on measures of statistical dependency it cannot tell either about the direction of these interactions or whether the interactions represent direct or polysynaptic effects. Effective connectivity captures the dynamical nature of neuronal interactions together with structural models to show how one neuronal population influences another. This talk will focus on the concept of effective connectivity, its relationship to structural and functional connectivity, its response to experimental context and currently available methods for its quantification including Granger causality and dynamic causal modeling.
The battle of the sexes (210.3)
Authors: Capel Blanche , Natarajan Anirudh , Maatouk Danielle , Munger Steven , Jameson Samantha
Published: June 2021

Although genetic sex is determined at fertilization, the decision to differentiate as a male or female occurs within the bipotential gonad during fetal life. Somatic progenitor cells in XX and XY mouse gonads are “primed” to differentiate along either the male or female pathway. At the bipotential stage, both XX and XY cells express very similar levels of genes associated with both ovary and testis development. The expression of the Y‐linked male sex‐determining gene (Sry) in XY gonads leads to both activation of male pathway genes, and repression of female pathway genes. Work using mutant and transgenic mice suggests that failure to suppress the female pathway is critical to establish male (testis) development. Bifurcation of organ fate begins between E11.0‐E12.0 in mice and involves a dynamic transcription network that is poorly understood. We profiled the XX and XY gonad transcriptomes at fine granularity during this period and conducted a DNase Hypersensitivity study to define promoters and enhancers. Our goal is to define the cascades of gene activation and repression that establish male or female development. This work is funded by a grant from NIH to BC.
Epithelial cell lineage specification during prostate organogenesis and regeneration (210.4)
Authors: Shen Michael , Wang Zhu , Shibata Maho , Talos Flaminia , Chua Chee Wai , Lei Ming , Bergren Sarah
Published: June 2021

The prostate gland is a particularly interesting system for the investigation of cell lineage specification given the central role of androgen signaling in all aspects of its development and function, as well as its ability to undergo serial regression and regeneration in response to androgen deprivation and restoration. In our previous studies, we have shown that a known regulator of prostate epithelial differentiation, the homeobox gene Nkx3.1, marks a luminal stem cell population that functions during prostate regeneration and is an efficient target for oncogenic transformation in prostate cancer. Genetic lineage‐tracing studies demonstrated that these rare cells which express Nkx3.1 in the absence of testicular androgens (castration‐resistant Nkx3.1‐expressing cells, CARNs) are bipotential and can self‐renew in vivo, and can serve as a cell of origin for prostate cancer. In parallel studies, we have utilized lineage‐tracing and transplantation approaches to explore the stem/progenitor properties of prostate basal cells during regeneration and tissue homeostasis, and observed an unexpected plasticity of basal cells in these assays. We will present our recent findings on prostate lineage specification and discuss their implications with respect to the relationship between epithelial progenitor populations and prostate cancer initiation.
Inflammation and the capacity for organ regeneration (212.1)
Authors: Mescher Anthony
Published: June 2021

Most vertebrate species used for studies of regeneration (amphibians, teleosts) are deficient in adaptive immunity compared to mammals. Recent data suggest that such immunodeficiency may be linked to regenerative capacity. The profile of cytokines, chemokines, and leukocytes elicited by limb amputation in regenerating amphibians is generally similar to that produced during the early phase of mammalian wound repair. Regeneration of limbs and tails requires production of reactive oxygen species, macrophage activity, and other components of inflammation. In the frog Xenopus laevis the ability to regenerate well‐patterned limbs is lost during the prometamorphic period. Analyses of gene expression in Xenopus limbs amputated at different developmental stages reveal changes in inflammation and resolution during the period when regenerative capacity declines. Nonregenerating limbs appear delayed in resolving inflammation, which does not impair wound closure or local cellular reprogramming but perturbs the regulated signaling events required for tissue patterning. Anti‐inflammatory agents improve regeneration in appendages amputated when regenerative ability is transiently or initially diminished. Data reviewed here support the hypothesis that development of adaptive immunity interferes with the patterning phase of organ regeneration and point to possible mechanisms for this effect.
Micromanaging inflammation and tissue repair (212.4)
Authors: Roy Sashwati
Published: June 2021

Inflammation is a protective response of the body to infection or injury. Given that miRNAs are fundamental to the post‐transcriptional control of gene expression, it is not surprising that role of specific miRNAs in the immune and inflammatory response has been discovered. The best characterized miRNA for inflammation are miR‐146, miR‐21 and miR‐155, all of these have been shown to be strongly induced in multiple cell types by proinflammatory stimuli following tissue injury. At an injury‐site, efficient clearance of apoptotic cells by wound macrophages or efferocytosis is a pre‐requisite for the timely resolution of inflammation. Emerging evidence indicates that miR‐21 may regulate the inflammatory response. The significance and mechanisms of miR‐21 in the regulation of efferocytosis mediated suppression of innate immune response, a key process implicated in resolving inflammation following injury will be elucidated.

Wound healing research in the author's laboratory is funded by NIDDK R01 DK076566 to SR.
Differential expression of corticotropin releasing factor and urocortins in the Schistosoma mansoni‐infected mouse ileum (214.2)
Authors: Buckinx Roeland , Bagyánszki Mária , Avula Leela , Adriaensen Dirk , Van Nassauw Luc , Timmermans Jean‐Pierre
Published: June 2021

Corticotropin releasing factor (CRF) and urocortins (UCNs) are important ligands in the CRF signaling pathways, most well‐known from the hypothalamic‐pituitary‐adrenal stress axis. However, peripheral CRF signaling has profound effects on gastrointestinal functions. Despite the potential of the murine animal model in elucidating this complexly balanced pathway through genetic manipulation, little is known regarding the expression of CRF and UCNs in the intestinal wall of the mouse. This study therefore investigates the cellular localization of CRF and UCNs in the ileum and examines how this cellular expression is altered in intestinal schistosomiasis. The results show a diverse expression of CRF and UCNs in the enteric nervous system, granulocytes and the ileal musculature. Of special interest was the de novo expression of UCN3 in a majority of submucous vasodilator/secretomotor neurons, possibly strengthening CRF2‐mediated effects during inflammation. In conclusion, we outline for the first time the ileal expression of CRF signaling ligands in the mouse, and point to important changes of this signaling system in intestinal inflammation.

Grant Funding Source: Supported by a grant of the Flemish Organization for Scientific Research (FWO G.0179.08)
Roles of glycosaminoglycans in brain physiology and neurological disorders (223.2)
Authors: Yamaguchi Yu
Published: June 2021

Glycosaminoglycans (GAGs) are a family of linear acidic polysaccharides, including heparan sulfate, chondroitin sulfate, keratan sulfate, and hyaluronan. GAGs are present mainly in the cell surface and extracellular matrix as proteoglycans. Physiological roles of GAGs in various tissues can be effectively studied by using conditional knockout of glycosyltransferases necessary for their synthesis. We have used this approach to determine the roles of GAGs in the nervous system. Earlier studies have established the essential roles of heparan sulfate in various aspects of brain development. More recently, we have applied this approach to investigate the role of GAGs in neurophysiology, cognition and behavior. This line of studies has revealed unexpected roles of GAGs in the pathogenesis of neurological and mental disorders. For instance, the removal of heparan sulfate from excitatory neurons results in striking recapitulation of the constellation of autism‐like behavioral phenotypes, which is correlated by reduced surface expression of glutamate receptors in excitatory synapses.
Deep methylomes reveal diverse roles of DNA methylation in human tissues (235.1)
Authors: Ecker Joseph , Schultz Matthew , He Yupeng , Hariharan Manoj , Whitaker John , Nery Joseph , Urich Mark , Chen Huaming , Leung Danny , Rajagopal Nisha , Lin Shin , Lin Yiing , Snyder Michael , Wang Wei , Ren Bing
Published: June 2021

In mammals, DNA methylation is a well‐known contributor to cellular identity and is required for normal development. A thorough mapping of the diversity of methylation patterns is a key step in understanding their role in establishing the identity of different tissues. We have produced high coverage methylomes, transcriptomes, and genomes of 18 different tissue types from 4 human donors. These data revealed a large set of differentially methylated regions, many of which are tissue‐specific and identify regions important for tissue function. In addition to locus specific changes, these methylomes revealed partially methylated domains in pancreas, which covered a significant fraction of its genome and are transcriptionally repressed. We found genome‐wide evidence for nonCG methylation in a subset of these tissues, a finding that had previously been restricted to embryonic stem cells and brain tissue. These results extend our knowledge of the unique role of DNA methylation in organ development and function, and offer a new framework for testing the role of the epigenome in healthy function and in pathological states in each of the major organ systems.
Systems analysis of mitochondrial pathogenesis (235.3)
Authors: Mootha Vamsi
Published: June 2021

Mitochondria lie at the heart of cellular metabolism and signaling. Recent genetic studies indicate that the organelle plays crucial roles both in rare, inborn errors of metabolism, as well as in some of the most common, age‐associated degenerative diseases. At present how lesions within mitochondria give rise to pathogenesis is not clear. In this talk I will present our recent work that combines human genetics with large‐scale genomic profiling to better understand mitochondrial pathogenesis. The results provide insight into new therapeutic targets for rare and common diseases.
Nuclear transport and gene expression (238.3)
Authors: Weis Karsten
Published: June 2021

Nuclear export is an essential step in the expression of every eukaryotic mRNA. However, it remains poorly understood how mRNAs are unidirectionally transported across the nuclear envelope and how this process is regulated. The DEAD‐box ATPase Dbp5/DDX19 functions in mRNA export and is thought to remove mRNA export factors from the message to terminate the translocation through the nuclear pore complex (NPC). Dbp5 is localized to the NPC via an interaction with Nup159/Nup214 and is locally activated there by Gle1 together with the small‐molecule inositol hexakisphosphate (IP6). Local activation of Dbp5 at the NPC by Gle1 is essential for mRNA export in vivo but the mechanistic role of Dbp5 in mRNP export remains unlcear. We have addressed the question of how Dbp5 contributes to directional mRNA export and have analysed the function of Dbp5 biochemically, structurally and by single molecule experiments.

Once mRNAs reach the cytoplasm, they can be either translated, stored or decayed. An important regulator of the transition between mRNA translation and decay is the DEAD‐box ATPase Dhh1/DDX6. We have shown that recruitment of Dhh1 to an mRNA is sufficient to move this mRNA from an active state to translational repression and to localize it into processing bodies (P bodies). Our research has focused on the question of how the ATPase function of Dhh1 is used to control whether an mRNA is translated, stored or decayed.
Fatty acid desaturase 3 null mouse biochemical phenotype (246.5)
Authors: Zhang Jiyao , Qin Xia , Liang Allison , Kim Ellen , Lawrence Peter , Kothapalli Kumar , Brenna J.
Published: June 2021

Introduction. We report here generation of the first Fads3 knockout (KO) mouse and initial observations about its biochemical phenotype. Fatty acid desaturase 3 (FADS3) is a member of the FADS gene cluster. FADS1 and FASDS2 code for the rate‐limiting enzymes in LCPUFA biosynthesis. FADS3 has no known function.

Methods. All mice (wild type (WT) and KO) were fed standard rodent chow devoid of LCPUFA. Brain and livers were harvested on day 1, 3, 7, 13, 21, and 30 of age for fatty acid and gene expression analysis. Gene expression levels were measured by quantitative and semi‐quantitative real time PCR.

Preliminary Results. No differences in overt phenotypes (survival, fertility, growth rate) were observed between WT and KO. Docosahexaenoic acid (DHA, 22:6n‐3) levels in the brain of postnatal day 1 (P1) KO mice were significantly (P<0.05) lower than the WT. No significant Fads1 and Fads2 mRNA expression changes were detected. Elovl2 is known to catalyze the conversion of C20 PUFA. Hepatic Elovl2 expression was significantly upregulated in P1 KO mice compared to WT, and the 22:6n‐3‐precursor 22:5n‐3 was about double in KO vs WT mice.

Conclusion. Fads3 null mice have lower brain 22:6n‐3, greater liver 22:5n‐3, and greater liver Elovl2 expression on Day1, though all levels normalize within days. Fads3 appears to influence liver‐mediated 22:6n‐3 synthesis and brain 22:6n‐3 accretion in perinatal life in the Fads3 KO mouse.

Grant Funding Source: NIH R01 AT007003
Iron and complementary feeding of breast‐fed infants (247.3)
Authors: Friel James , Qasem Wafaa , Hossain Zakir , Jorgensen Sarah
Published: June 2021

The American Pediatric Society and Health Canada both recommend that iron fortified cereal (most iron unabsorbed) or meat (most iron absorbed) be introduced to all breast‐fed infants at six months of age to support iron stores and linear growth. Evidence suggests that the iron contained in these foods, particularly cereal, is not fully absorbed and will collect in the colon with the possibility of free radical generation and intestinal inflammation.

Objectives: To assess the safety of the recommended first solid food for exclusively breast‐fed infants from a free radical perspective.

Methods: Ninety exclusively breastfed infants are being randomized to 1 of 3 feeding groups: iron fortified cereal (FeCer), iron fortified cereal with fruit (FeCF) or meat (M). Urine and stool samples are collected before introduction of study foods (4‐5.5 months) and 3 weeks after introduction of these foods to assess: urinary F2 isoprostanes (LC‐MS‐MS), urinary 8OH‐deoxy guanosine, fecal calprotectin (ELISA), reactive oxygen species (ROS‐HPLC) and non‐heme iron generation (Colorimetric) in the stool, and the fecal microbiome by 16S RNA gene pyrosequencing.

Preliminary results are: urinary isoprostanes (base: 10.1+7.2; after feeds 12.8 + 7.6 ng/mg/ml creatinine; n=25) did not differ over time or between feeding groups at either baseline or after feeds. ROS did not differ between groups at either sampling time nor within the FeCF and M group over time, however there was a trend to increased free radical generation in the FeCer group (base: 0.018 + 0.019; after feeds 0.037 + 0.015 RU; n=12; P = 0.087).

Conclusion: Our hypothesis was that iron fortified cereals would provide unabsorbed iron that would reduce the ability of the colon to resist oxidative stress and has been supported from this preliminary subset. The newborn breast‐fed infant may not cope with large amounts of unabsorbed iron in the colon.

Grant Funding Source: Supported by CIHR
4‐Hydroxynonenol is present in human milk and related to gestational age at delivery and excessive infant weight gain (247.6)
Authors: Young Bridget , Morrow Ardythe , Davidson Barbara , Geraghty Sheela , Patinkin Zachary , Krebs Nancy
Published: June 2021

4‐hydroxynonenol (HNE; a protein adduct product of lipid peroxidation) may serve as a novel marker of oxidative stress in human milk (HM). We investigated the determinants of HNE in HM and relationships between HNE and maternal BMI, HM cytokines, and infant growth.

Anthropometry was performed on 36 mothers and their healthy term (37‐42 weeks) exclusively breastfed infants at T1 (2 weeks), T2 (4 months) and T3 (6 months). HM was collected at T1 and T2 for macronutrient, HNE, IL‐6, IL‐8, and TNF‐α analysis. Infants gaining more than 0.66 weight‐for‐length Z‐scores from T1 to T3 were considered over‐gainers.

HNE in HM decreased from T1 to T2 (p<0.01); this decrease was attenuated when controlling for total protein in HM (HNE:PRO) (p=0.09). HNE concentrations at T1 were positively associated with those at T2 (p=0.02, R2=0.16). Gestational age at delivery was inversely associated with HNE:PRO (p=0.02; R2=0.16). HNE:PRO (but not unadjusted HNE) was higher in HM of female infants at T1 (p=0.02) and T2 (p=0.09). HNE in HM was not related to maternal BMI or HM cytokine or macronutrient composition. HM of over‐gainers at T2 exhibited a trend for higher HNE:PRO (p=0.095) (but no difference in unadjusted HNE).

These preliminary results suggest that HNE in HM may be impacted by length of gestation. HM HNE may affect infant weight gain patterns. Further research is warranted to fully characterize effects of this pro‐oxidant marker.

Grant Funding Source: Supported by NIH T32 DK007658‐21, 5K24DK083772; Thrasher Research Fund; University of Colorado CCTSI
Identification of structural features of ursolic acid required to protect monocytes against metabolic stress‐induced dysfunction (249.5)
Authors: Nguyen Huynh Nga , Asmis Reto
Published: June 2021

We showed that ursolic acid (UA), a pentacyclic triterpenoid with anti‐inflammatory properties, reduces atherosclerotic lesion size and improves kidney function in diabetic mice. We proposed that the atheroprotective effects of UA are due to its ability to protect monocytes against metabolic stress‐induced dysfunction or “priming”, i.e. the Nox4‐mediated hyper‐responsiveness to chemokine‐induced cell adhesion and migration. The aim of this study was to test eight of UA’s naturally occurring structural analogs and identify the structural features of UA required to inhibit monocyte priming. We determined in metabolically primed THP‐1 monocytes the doses (0.03 ‐ 3 μM) at which UA and its analogues blocked metabolic stress‐induced Nox4 induction and accelerated chemotaxis in response to MCP‐1. Only the UA analogs uvaol and corosolic acid protected monocytes against metabolic priming. We found that altering the position of one methyl group in position 19 of UA abolishes its protective effects. Furthermore, the carboxyl group in position 17 appears to be critical but not sufficient for UA to prevent monocyte priming by metabolic stress. In summary, our data suggest a highly selective molecular mechanism for the protective effects of UA on monocyte priming and dysfunction and UA’s anti‐atherogenic properties. Identifying the functional groups responsible for UA’s beneficial effects will allow us to develop more potent, low‐cost dietary supplements for the prevention of atherosclerosis and other monocyte‐mediated diabetic complications.

Grant Funding Source: Supported by grants to RA from the NIH (R01‐AT006885)
Wheat phenolics can prevent inflammation in human umbilical endothelial cells: effects on biomarkers relevant to development of cardiovascular diseases (249.7)
Authors: Noratto Giuliana , Mohanty Indira , Alcantara Zapata Diana , Chew Boon
Published: June 2021

Cardiovascular disease (CVD) is the number one cause of death around the world. Whole wheat is a source of phenolics that can protect against inflammation‐induced CVD. However, the contents of extractable (EP) and non‐extractable (NEP) phenolics among cultivars and potential for CVD prevention have not been investigated.

We assessed EP and NEP in 7 wheat cultivars grown in Washington and their protective effects (2‐8 µg/mL ) on modulation of glutathione S‐transferase activity (GST) by spectrophotometry. Reactive oxygen species (ROS) and expression of inflammatory genes in HUVEC challenged with E. Coli lipopolysaccharides (LPS)(2µg/mL) or glucose (G)(25 mM) were assessed with commercial kits and qPCR.

Results showed that EP contents were similar (0.7‐0.8 mg GAE/g) while NEP varied significantly (0.5‐1 mg GAE/g) among cultivars. LPS challenge induced ROS in HUVEC by 1.4‐fold of control and this effect was prevented by WestBred936 (spring hard) wheat phenolics in a dose‐dependent manner up to levels found in controls not challenged with LPS. Similar trend was found in G‐challenged cells. This protection was in part due to the induction of GST up to 2‐ and 3.5‐fold of LPS‐ and G‐challenged cells, and due to downregulation of NF‐ĸB mRNA levels and NF‐ĸB‐target genes IL‐6, ICAM1, and VCAM1. We are currently validating these results in vivo. Overall, these results may be of clinical relevance in the mitigation of inflammatory conditions that mediate the onset of CVD.

Grant Funding Source: Supported by Washington Grain Commission
A tailored, fortified, poultry‐based spread (Spammy™) improves vitamin D and B12 of Guatemalan preschoolers as part of an institutional menu (250.5)
Authors: Hernandez Liza , Salguero Sylvana , Bonorden Melissa , Herreid Richard , Armas Laura , Paul Ligi , Selhub Jacob , Solomons Noel
Published: June 2021

Background: Although the early years of life inherently pose nutritional vulnerability, geographical and cultural determinants dictate distinct patterns of nutrient deficiencies. Thus, tailoring responses holds the promise of more efficient and comprehensive redress.

Objective: To assess the efficacy of a poultry‐based spread, fortified with between 20% and 333% of the RDA of 12 selected micronutrients, including vitamins D and B12, both shown to be precarious among Guatemalan preschoolers in a previous survey.

Methods: A total of 167 preschoolers, aged 4‐7 y were randomized to receive 43 g of fortified (FS) or unfortified (US) SpammyTM during weekdays over 20 wks as part of the institutional breakfast at a faith‐based day‐care center on the outskirts of Guatemala City in a triple‐blinded format. Circulating nutrient status indicators (ferritin, 25(OH)vit D, vitamin B12) were measured at both ends of the trial.

Results: Over 75% of the offered spread was consumed on 84% of serving days with no difference between treatment groups. At baseline, blood biomarkers were abnormally low for iron in 20%, vit D in 59% and vit B12 in 19%. A significant 9% increase in vit D (p<0.0001) and 23% vit B12 (p=0.0017) with FS, unmatched in the US group, were seen.Conclusion: Regular consumption of a fortified spread during half of the school year support improvement in the status of two nutrients problematic for Guatemalan preschoolers.

Grant Funding Source: Supported by McGovern‐Dole Food for Education and Child Nutrition Program and Hormel Foods
A program impact pathway‐based process evaluation helps trace program implementation processes essential to achieving changes in infant and young child feeding practices in the Alive & Thrive initiative in Bangladesh (251.4)
Authors: Saha Kuntal , Rawat Rahul , Khaled Adiba , Kennedy Andrew , Roopnaraine Terry , Bhuiyan Mahbubul , Kim Sunny , Ruel Marie , Menon Purnima
Published: June 2021

A&T aims to improve IYCF practices through intensive interpersonal communication (IPC) by well‐trained frontline workers (FLWs) and a mass‐media (MM) campaign. A cluster‐randomized impact evaluation in 20 subdistricts comparing A&T intensive [(A&T‐I) (IPC+MM)] and A&T non‐intensive [(A&T‐NI) (MM only)] areas shows positive early impacts of A&T‐I interventions (Menon et al.2013). A PIP‐based PE was conducted to examine 5 domains: FLW training; service delivery; coverage; service utilization; and behavioral impacts. Data: Surveys of randomly selected households with children 0‐23 mo (N=1000 per group) in 2010 (baseline) and 2013; 90 in‐depth qualitative interviews. At baseline, there were no differences in any domains. In A&T‐I areas in 2013, IYCF knowledge of FLW was better; FLW home visits were higher (89% vs. 40% in last 1 mo; p<0.05); and more mothers received IYCF advice from FLW (92% vs. 50%; p<0.05) compared to A&T‐NI areas. Maternal recall of 6 MM television spots in A&T‐I areas was 45‐65% and was slightly higher than in A&T‐NI areas (n.s.). Maternal IYCF knowledge, particularly of breastfeeding and introduction of complementary foods was better (p<0.05) in A&T‐I areas. Qualitative data shows that some negative social norms still persist and that infants were rarely fed adequate diets throughout the first 2 y of life. This PIP‐based PE indicates that key program elements of coverage, quality of service and intervention reach are in place, but some household‐level factors could limit impact.

Grant Funding Source: Funding: Bill & Melinda Gates Foundation, through Alive & Thrive, managed by FHI360.
The growing dominance of Walmart in U.S. consumers’ food shopping baskets and caloric purchases: trends from 2000‐2012 (255.2)
Authors: Smith Lindsey , Ng Shu , Popkin Barry
Published: June 2021

Little is known about who shops at Walmart, the proportion of calories purchased there, or kcal/dollar spent relative to other food stores. Using Nielsen Homescan pooled cross‐sectional nationally representative data on household purchases, we examined changes in annual percent of calories of consumer packaged foods/beverages (CPGs) purchased at Walmart from 2000‐2012 (N=643,600). The percent of households who shopped at Walmart rose from 67% to 78% from 2000 to 2012 (p<0.01). From quantile regression, among Walmart shoppers, the percent of CPG calories purchased at Walmart increased from a median of 4% in 2000 to 15% in 2012 (p<0.01). This change was even greater at the upper end of the distribution, with the 75th percentile of Wal‐Mart shopper purchasing 11% of CPG calories at Wal‐Mart in 2000 compared to 45% in 2012 (p<0.01). From multinomial logistic regression, 54% of low‐income households purchased a high proportion of their calories from Walmart compared to 35% of high‐income households (p<0.01). Race‐ethnic minority subpopulations were less likely to be in the high shoppers group compared to Whites (p<0.01). Walmart showed a greater decline in kcal/dollar spent than other food stores, decreasing from 734k kcal/dollar in 2000 to 441kcal/dollar in 2012 (p<0.01). Walmart plays an increasingly important role in influencing US food store purchases, particularly amongst low‐income and white subpopulations.

Grant Funding Source: Supported by the Robert Wood Johnson Foundation (68793) and the Carolina Population Cen
Challenges and facilitators for implementing prenatal calcium supplementation to prevent preeclampsia: formative research with health care providers and women in Kenya (256.1)
Authors: Dickin Katherine , Seim Gretchen , Young Sera , Wawire Salome , Martin Stephanie
Published: June 2021

Prenatal calcium supplementation is now recommended by the WHO to prevent preeclampsia and eclampsia (PE/E), a leading cause of maternal mortality globally. To guide calcium supplementation programming, we conducted in‐depth interviews with 20 Kenyan health care providers and 22 pregnant women/new mothers. Interviews assessed perceptions of PE/E, antenatal care (ANC), and challenges and motivations for delivering and taking supplements such as iron‐folic acid. Interviews were analyzed thematically. Most health staff were aware of the symptoms and severity of PE/E but saw it rarely. Women had not heard of PE/E and very few knew of any danger signs. Swelling of lower limbs was viewed as normal in pregnancy, as were many symptoms of anemia. Due to confusion about “low blood” (anemia) and “presha” (high blood pressure or PE/E), even among some health workers, messages promoting pills to prevent these two conditions appeared contradictory. Adherence barriers included lack of access to supplements and information about benefits, and low ANC attendance prior to late pregnancy. Widespread concern about poor diet quality in pregnancy was expressed. While respondents stated that this would motivate women to take calcium supplements, the lack of perceived risk of PE/E suggests a need for extensive staff training and community mobilization if this efficacious measure is to achieve its potential public health impact.

Grant Funding Source: Supported by The Micronutrient Initiative
Height and BMI at five years of age following prenatal supplementation with docosahexaenoic acid in Mexico (256.8)
Authors: Gonzalez‐Casanova Ines , Stein Aryeh , Hao Wei , Feregrino Raquel , Romieu Isabelle , Barraza‐Villarreal Albino , Rivera Juan , Martorell Reynaldo , Ramakrishnan Usha
Published: June 2021

Prenatal supplementation with docosahexaenoic acid (DHA) improves birth outcomes such as preterm delivery and birth size, but less is known if these differences translate into improved postnatal growth. We examined growth patterns through age 5 y in a cohort of 802 Mexican children (82% of the original cohort) whose mothers participated in a randomized controlled trial of daily supplementation with 400 mg/day of DHA or a placebo from week 18‐22 of gestation through delivery. Overall, means (standard deviation) of height for age and body mass index (BMI) for age z‐scores relative to the WHO growth reference were ‐0.49 (0.91) and 0.13 (1.11) respectively. Intent‐to‐treat analyses revealed no significant differences by treatment group (DHA‐placebo, mean, 95% confidence interval) for height (‐0.01, ‐0.42, 40 cm), or BMI (‐0.02, ‐0.26, 0.22 kg/m2) at age 5. Post‐hoc analysis revealed significant (p=0.001) heterogeneity by breastfeeding status for height: among children not breastfed at 3 months of age, those exposed to DHA were 2.20 cm (95% CI 0.62, 3.79) taller at age 5 than those in the placebo group; among children receiving mixed or exclusive breastfeeding at 3 months there was no effect of DHA supplementation on size at age 5. This same heterogeneity was observed at age 4 y but not at younger ages. Prenatal DHA supplementation may have differential long‐term effects on linear growth of children depending on breastfeeding history.

Grant Funding Source: Supported by NIH grants 5R01HD043099‐06 and 5R01HD058818‐04
Dietary magnesium is positively associated with indices of skeletal muscle mass and may attenuate the association between C‐reactive protein and muscle mass (257.2)
Authors: Welch Ailsa , Kelaiditi Eirini , Jennings Amy , McGregeor Alexander , Spector Tim , Cassidy Aedin
Published: June 2021

Background: Chronic age‐related muscle loss is one component of sarcopenia and dietary magnesium (Mg) intake may influence low‐grade inflammation, a risk factor for muscle loss. Therefore, associations between indices of skeletal muscle mass in women, and the potential attenuation by Mg of the relationship between C‐reactive protein (CRP) and muscle were investigated.

Methods: A cross‐sectional study of 2570 women aged 18‐79 years with fat free mass (FFM) measured using dual‐energy X‐ray absorptiometry, and with circulating HsCRP in 1658 participants. Dietary Mg was calculated using an FFQ and indices of FFM (FFM, as a percentage ‐ FFM% and FFM index ‐ FFMI ‐ FFM in kg/height2) calculated according to quintile of Mg using multivariate regression, and adjusted for age, physical activity, smoking, energy intake, anti‐inflammatory medication, HRT (for FFMI with BMI).

Results: Significant positive associations were found between Mg and FFM% (P trend =0.004) and FFMI (P trend=0.001) with extreme quintile differences of 1.58% for FFM% and 0.5 kg/m2 for FFMI. The association between indices of muscle mass and CRP (per quintile of CRP) was ‐1.2 % P<0.001 for FFM% and ‐0.07 kg/m2 for FFMI which was attenuated by Mg by 0.7 % for FFM% and 5.4% for FFMI.

Conclusions: We found significant positive associations between Mg and indices of muscle mass, and dietary Mg attenuated the associations between CRP and muscle mass. Dietary intervention studies are needed to confirm this finding.
Homocysteine and bone mineral density in older females in the United States (257.5)
Authors: Bailey Regan , Looker Anne , Gahche Jaime , Mills James , Weaver Connie
Published: June 2021

Previous studies suggest that elevated total homocysteine (tHcy, >13 µmol/L) is a risk factor for bone fracture and osteoporosis; but the data are equivocal and no mechanism has been demonstrated. Elevated tHcy can be caused by impaired renal function, genetic factors, or suboptimal B‐vitamin status. We examined the relationship between tHcy and total body and lumbar spine bone mineral density (BMD) in females 蠅50 years participating in NHANES 1999‐2004(n=2,923). These years had concurrent measurement of tHcy, MMA, RBC folate, serum B‐12, and whole‐body dual‐energy X‐ray absorptiometry. Elevated tHcy was significantly associated with lower total and lumbar BMD when compared to normal tHcy (p <0.001) before and after controlling for age, race/ethnicity, kidney function, BMI, physical activity, intakes of calcium and vitamin D and use of estrogen/progesterone. For example, females with elevated tHcy were more likely to have lumbar spine osteoporosis when compared to females with normal tHcy [OR 2.22 (95%CI:1.3,3.9), p <0.001]. Serum MMA, vitamin B‐12 and RBC folate were not significantly related to BMD. In this population of older females, tHcy appears to be an independent risk factor for low total body and lumbar spine BMD. The role of genetic and other factors that may elevate tHcy concentrations and adversely affect BMD should be explored.

Grant Funding Source: Supported by the Office of Dietary Supplements
Correlation between cola consumption and bone mineral density among Puerto Ricans living in the greater Boston area (257.7)
Authors: Jamal‐allial aziza , Tucker Katherine
Published: June 2021

Correlation between Cola Consumption and Bone Mineral Density among Puerto Ricans Living in the Greater Boston Area Aziza Jamal‐Allial1,Katherine L. Tucker21Health Sciences, Bouvé College of Health Sciences; Northeastern University. Boston, MA. 2Clinical Laboratory & Nutritional Sciences, University of Massachusetts Lowell, Lowell, MA.

Several studies have investigated the relationship between the consumption of soft drinks and bone mineral density (BMD), but results are conflicting. We aimed to study the effect of soft drink consumption on BMD at 4 hip sites in 273 men and 698 women in the Boston Puerto Rican Osteoporosis Study using dual‐energy X‐ray absorptiometry (DXA). Soft drink intakes were captured by food frequency questionnaire (FFQ). The mean intake of soft drinks was 2.2 ± 4.3 and 1.5 ± 3.0 servings/week, for men and women, respectively. Total intake of cola beverages was significantly associated with lower BMD at the total hip (β= ‐0.004, P‐value 0.002), trochanter (β= ‐0.003, P‐value 0.03) and femoral shaft (β= ‐0.01, P‐value 0.0005), but not at the femoral neck. Intake of diet‐cola was also significant at the femoral shaft and approached significance at the total hip and trochanter. In contrast, non‐cola soft drinks were not significantly associated with any hip measure. Our models were adjusted for age, sex, BMI, height, calcium‐vitamin D supplement use, dietary calcium intake, smoking, alcohol use, total energy intake, physical activity, and season of BMD measurement. In conclusion, the effect of cola, but not other soft drinks suggests that phosphoric acid may have a negative effect on bone status

Grant Funding Source: This study was supported by the National Institutes of Health grants P01 AG023394, P50 HL105185 and
Bone metabolism and renal stone risk during bed rest for men and women (257.8)
Authors: Morgan Jennifer , Heer Martina , Hargens Allen , Macias Brandon , Hudson Edgar , Shackelford Linda , Zwart Sara , Smith Scott
Published: June 2021

Bed rest is a model to simulate and study the human physiological adaptations to spaceflight, for example changes in bone metabolism and renal stone formation. On Earth, the incidence of osteoporosis in women is about double that of men while the incidence of stone formation is more likely in men. These sex specific differences lead to logical questions about the existence of sex‐specific effects during spaceflight that can be tested in the bed rest model. In this study, we combined data from five bed rest studies (n=50 men, 24 women) ranging in duration from 14 ‐ 90 days to assess sex differences in bone loss, bone biochemistry, and renal stone biochemistry. Prior to bed rest, men had higher bone mineral density and content (P < 0.001), and excreted 30 % more biomarkers of bone resorption (i.e., collagen crosslinks) and 20 % more calcium per day than did females (P <0.05). These offsets remained throughout the duration of the bed rest periods. Urine renal stone biochemistry was significantly altered for both men and women during bed rest, resulting in men having an increased renal stone risk compared to the general population. Actual renal stone risks in these studies were reduced by maintaining a high amount of fluid intake (>2.5 L/d). These results indicate that overall both men and women are equally affected by the skeletal unloading of bed rest.
Supplementation with leucine does not prevent development of obesity in rats fed a high fat diet (258.5)
Authors: Smith Dameon , Washington Tyrone , Davis Gina , Williams Derrick , Baum Jamie
Published: June 2021

Excess dietary fat consumption has been implicated in the development of obesity and diabetes. Obesity can be characterized by a disproportionate increase in fat mass. However, if muscle mass can be increased or maintained in obesity, this may facilitate weight loss. Historically, supplementation with the branched‐chain amino acid leucine has been shown to increase muscle protein synthesis via the protein kinase mTORC1. Recent studies suggest that supplementation with leucine also has the potential to reduce weight gain and fat deposition in high‐fat fed, obese mice. The objective of this study was to determine if long‐term dietary leucine supplementation prevents development of obesity in rats meal‐fed a high fat (60%) diet. Male, Sprague‐Dawley rats (n=30/dietary treatment) were meal‐fed (3 meals/day) either a control diet (C), control+leucine (CL), high‐fat (HF), or high‐fat+leucine (HFL) for 42 days. On day 42 rats were sacrificed at 0, 30 or 90 minutes postprandial. HF/HFL‐fed rats gained, 28 g more than rats fed C/CL diets (p<0.05). Insulin AUC was 35% higher in CL vs C and 13% higher in HFL vs HF. There was no difference in blood glucose values between treatment groups. Rats supplemented with leucine had significantly higher fat mass and significantly (p<0.05) lower muscle mass compared to their respective controls. However, preliminary data suggest that L and HFL activate translation initiation in muscle via 4E‐BP1 phosphorylation. Taken together, this study demonstrates that supplementation with leucine increases translation initiation, however this does not result in increased muscle mass. Supplementation with leucine does not prevent development of obesity in HF‐fed rats.

Grant Funding Source: Supported by Arkansas Biosciences Institute.
Mechanistic studies in swine to evaluate newborn infant vitamin A dosing strategies (260.1)
Authors: Tanumihardjo Sherry , Gannon Bryan , Riabroy Napaporn , Heying Emily , Davis Christopher , Grahn Michael
Published: June 2021

Clinically, infant care is challenging especially in resource‐poor developing countries where small‐for‐gestational‐age is common and in any country when an infant is born prematurely. Several components comprise perinatal mortality of infants. Children suffering from vitamin A deficiency have a greater risk of irreversible blindness and dying from infectious diseases. Supplementation programs are an approach to address this issue. Studies have found little or no improvement in infant vitamin A status after supplementing with 25,000 IU vitamin A. Based on WHO’s recommendations, children ages 6 ‐ 11 mo receive 100,000 IU vitamin A and those 12 ‐ 59 mo receive 200,000 IU vitamin A every 4 to 6 mo. Multi‐centered and randomized trials of infant supplementation programs have shown inconsistent results. Infant supplementation itself has been debated due to the higher incidence of bulging fontanelle in vitamin A‐dosed infants. Therefore, a series of studies were performed in a swine model to evaluate infant dosing regimens. Newborn piglets from vitamin A‐depleted sows received a placebo, 25,000, 50,000, or 200,000 IU vitamin A supplements at birth. Tissues were collected and analyzed. The degree of vitamin A deficiency did not seem to affect the response to treatment. High doses may be quickly catabolized to maintain balance. Lung and spleen require chylomicron vitamin A to maintain concentrations, which was confirmed with a tracer study using α‐retinol. The tracer study also showed quick uptake in the adrenal gland. Low birth weight piglets took up less vitamin A. Food sources of vitamin A performed better than supplementation at improving vitamin A status of offspring.

Grant Funding Source: Supported by the Nutrition Unit at the World Health Organization.