bluuebunny/crossref_metadata_2025 huggingface dataset:
DHA‐enriched fish oil modulates B cell function and modifies clustering of lipid microdomains
Authors: Gurzell Eric Anthony , Teague Heather , Harris Mitchel , Clinthorne Jonathan , Shaikh Saame Raza , Fenton Jenifer
Published: June 2021

Fish oil has significant immunomodulatory effects and is primarily associated with suppression of T lymphocyte and dendritic cell function. The purpose of this study was to investigate the changes that DHA‐enriched fish oil (DFO) feeding elicits on B cells, an understudied, yet important cell population with respect to inflammatory pathologies. Utilizing confocal microscopy and flow cytometric analyses of purified splenic B cells, we assessed the effects of DFO on B cell activation in 129Sv mice (n=10/treatment). DFO feeding altered the lipid composition of splenic B cells and diminished GM1 microdomain clustering in the membrane. The clusters on B cells from mice fed the DFO diet appear 0.275 μm larger compared to control fed mice. In vivo, DFO increased lymphoid tissue B cell populations and surface markers of activation compared to controls. Ex vivo, LPS‐stimulated B cells isolated from DFO fed mice expressed more CD40 and produced more IL‐6 (50% increase) and TNF‐α (40% increase) than those from control mice, however OVA‐FITC antigen uptake was similar. DFO fed mice had significantly higher plasma IL‐5, IL‐13, and IL‐9 (Th2‐biasing cytokines) and cecal IgA compared to control. These results support the hypothesis and an emerging concept that fish oil enhances B cell function in vivo. These data are consistent with observations in C57Bl/6 mice, suggesting dietary fish oil enhances humoral immunity.

Grant Funding Source: NIH R03CA162427 (to J.I.F.) and NIH R15AT006122 (to S.R.S.)
Relationship Between In Vitro Absorption of Chondroitin Sulfate (CS) and Its Molecular Weight
Authors: Surapaneni Lahari , Haley‐Zitlin Vivian , Bodine Ashby , Jiang Xiuping , Brooks James
Published: June 2021

Objective

The purpose of this study was to investigate if the molecular weight of chondroitin sulfate (CS) has an impact on its absorption in vitro, in a Caco‐2 cell model.

Methods

Caco‐2 cells in 24 well plates were incubated for 2 hours with eight CS samples of differing molecular weights (7 kDa – 35 kDa). The amount of CS transported into the basolateral side of the Caco‐2 monolayer was quantitatively determined to calculate the permeability coefficients (Papp).

Results

Of the 8 samples evaluated, four samples had a Papp value of 15 × 10−6 cm sec−1 or higher indicating moderate to high absorption. Two of the four samples with higher Papp values were high molecular weight compounds (~ 35 kDa).

Conclusion

Within the molecular weight range used in this experiment, the absorption of CS samples did not have a correlation with their molecular weights but, interestingly, correlation was observed between the absorption and percentage of 6‐sulfated disaccharide in the CS samples.

Grant Funding Source: Pharmavite, Inc.
Liquid and solid sources of added sugar and their associations with body weight and metabolic syndrome components in children
Authors: Wang Jiawei , Light Kelly , Johnson‐Down Louise , St‐Arnaud‐McKenzie Danielle , O'Loughlin Jennifer , Paradis Gilles , Gray‐donald Katherine
Published: June 2021

Limited evidence exists in children relating added sugar (ASug) from solid‐food sources with obesity and metabolic syndrome (MetSyn) in contrast to considerable data on sugar‐sweetened beverages (SSB). Our aim was to examine dietary sources of ASug and their relationship with body weight indicators and MetSyn components. Exposure data were three 24‐hour dietary recalls from 613 Caucasian children (8–10 y) in the QUebec Adiposity and Lifestyle InvesTigation in Youth (QUALITY) study (2008–2010). Outcome measurements included body mass index (BMI), waist circumference (WC), waist‐to‐height ratio (WHR), triglyceride, HDL‐C, systolic blood pressure (SBP) and HOMA‐IR. Multivariate regressions were used with covariates of age, sex, fat mass index (fat mass by dual‐energy absorptiometry) and physical activity (7‐day accelerometer). On average, 12% of energy intake (204 kcal) came from ASug. Four top sources of ASug were sweets (contributed 28% of ASug), baked products (25.9%), SSB (17.1%) and ready‐to‐eat cereal (5.6%). No associations were found with overweight or metabolic indicators with solid‐food sources of ASug. This was in contrast to the association of ASug from SSB with BMI, WC, WHR and SBP. Although children consumed more ASug from sweets and baked products than from SSB, these solid sources of ASug were not associated with health indicators.

Grant Funding Source: Canadian Institutes of Health Research, Heart and Stroke Foundation of Canada and Fonds de la recherche en sante du Quebec.
Salmonella Typhimurium Directs the Localization of the Desmosomal Protein, PERP, to Induce Inflammation
Authors: Hallstrom Kelly , Casanova James E , McCormick Beth A
Published: June 2021

Salmonella enterica serovar Typhimurium is one of many Salmonella strains responsible for over 40,000 cases of Salmonella‐induced gastroenteritis in the U.S. yearly. Salmonella invade the apical surface of intestinal epithelial cells and trigger inflammation primarily through the activity of the type three secreted effector, SipA, which activates signal transduction cascades that ultimately lead to the recruitment of neutrophils (PMN) to the site of invasion. Previous studies showed that SipA acts extracellularly at the apical surface suggesting it interacts with an apically‐located host protein. Recently, we identified the desmosomal protein, PERP, as a SipA‐binding partner. PERP accumulates at the apical surface in response to Salmonella in a SipA‐dependent manner and is a key regulator of transepithelial PMN migration during Salmonella infection. In examining the mechanism by which this occurs, we noted that pharmacologic inhibition of the endocytic pathway by the drug dynasore blocks PERP re‐internalization. Sucrose gradient results also showed that during Salmonella infection PERP is enriched in the apical membrane and the endosomal fractions, suggesting that PERP is trafficked to the apical surface using the endosome recycling system. We propose that S. Typhimurium interferes with the homeostatic re‐uptake of PERP, which is governed by the endosome recycling pathway, via the activity of SipA.

Funding Sources: NIH, NIDDK, Crohn's and Colitis Foundation of America
Fundamental role for HIF‐1α in expression of enteric human β defensin‐1
Authors: Kelly Caleb James , Glover Louise E , Campbell Eric L , Ehrentraut Stefan F , Kominsky Doug J , Bowers Brittelle E , Bayless Amanda J , Saeedi Bejan J , Colgan Sean P
Published: June 2021

Antimicrobial peptides are secreted by the intestinal epithelium to defend from microbial threats. The antimicrobial activity of human β defensin‐1 (hBD‐1) is potentiated by this low‐oxygen environment and the hBD‐1 gene (DEFB1) is constitutively expressed in intestinal epithelial cells (IEC). As previous studies have indicated that hypoxia‐inducible factor (HIF) is active even in healthy intestinal mucosa, we compared the transcriptional profile of IEC in mice lacking IEC HIF‐1α to those of wild‐type mice. This unbiased screen identified a key role for epithelial HIF‐1α in expression of murine defensins. Studies with human IEC revealed that basal HIF‐1α is fundamental for the constitutive expression of hBD‐1 mRNA and protein. While hypoxia did not induce DEFB1 expression, attenuated basal HIF‐1α expression (ascorbate supplementation) repressed DEFB1, suggesting a low threshold for HIF‐1α saturation. DEFB1 promoter studies confirmed HIF‐1α binding to a hypoxia response element. Since DEFB1 is unique to humans, we used 94 human intestinal samples to reveal a strong correlation between DEFB1 and the canonical HIF‐1α target gene GLUT1. Taken together, these findings indicate that basal HIF‐1α is critical for constitutive expression of DEFB1 in the intestine and supports efforts that target HIF in restoration and maintenance of intestinal integrity.

Research Support:

F30DK096709, TL1RR025778, DK50189, HL60569, DK095491 and by the Crohn's and Colitis Foundation of America.
Cleavage of interleukin‐8 and attenuation of neutrophil chemotaxis by a Giardia cathepsin B
Authors: Cotton James , Bhargava Amol , Ferraz Jose , Hollenberg Morley , Beck Paul , Buret Andre
Published: June 2021

Parasitic immunomodulatory molecules are currently undergoing investigation for their potential as anti‐inflammatory therapeutics. Children infected with the small intestinal parasite Giardia duodenalis have attenuated inflammatory responses via unknown mechanisms.

Objective

To determine if Giardia trophozoites modulate interleukin‐8 (CXCL8) secretion from intestinal epithelial cells (IECs) resulting in attenuated neutrophil (PMN) chemotaxis.

Results

Co‐incubation of Giardia trophozoites with ex vivo small intestinal mucosal biopsy tissues or in vitro monolayers resulted in attenuation of IL‐1β‐ or S. typhimurium‐induced CXCL8 secretion via parasitic cleavage of CXCL8. Giardia‐mediated cleavage of CXCL8 decreased its chemotactic ability for PMNs. Giardia trophozoites secrete cysteine proteases in the presence or absence of in vitro monolayers. Inhibition of Giardia cysteine proteases with a cysteine protease inhibitor (E‐64d) or a cathepsin B‐specific inhibitor (Ca‐074Me) inhibited Giardia‐mediated cleavage of CXCL8.

Conclusion

Giardia trophozoites release a cathepsin B‐like protease that cleaves CXCL8 and attenuates PMN chemotaxis. Funding provided by NSERC, AIHS, and CCFC.
MUC2 exocytosis in intestinal goblet cells
Authors: Cornick Steve , Moreau France , Chadee Kris
Published: June 2021

MUC2 in the major secreted gel‐forming mucin produced by goblet cells (GC) that form the protective mucus blanket covering the epithelium as the first line of innate host defense in the gut. The protein complex and the cellular mechanisms by which MUC2 granulae are released by compound exocytosis from GC under basal or in response to mucus secretagogues are not known. In this study we identified the putative exocytosis components expressed in human GC, LS 174T. The exocytosis protein complex formation between SNAP23, Syntaxin 3 and Munc18b was studied in response to the PKC agonist, phorbol 12‐myristate 13‐acetate (PMA) using immunoprecipitation and mass spectrometry. In contrast to MUC2, the exocytosis proteins were not regulated at the transcriptional level, as mRNA and protein expression was unaltered after PMA. However, PMA treatment phosphorylated Syntaxin 3 in active SNARE complex formation. Surprisingly, confocal imaging revealed that Syntaxin 3 and Munc18b are not only localized to the apical cap of the GC theca but also to areas around the theca where the SNARE components co‐localized with actin filaments. These findings were corroborated using differential centrifugation where Syntaxin 3 was identified on both the granulae and the plasma membrane. Our studies reveal complex exocytosis protein machinery that regulates both basal and stimulated MUC2 granulae release from goblet cells. Supported by CIHR.
αSNAP controls β1‐integrin trafficking and FAK/Src dependent cell‐matrix adhesions in human epithelial cells
Authors: Naydenov Nayden Gueorguiev , Feygin Alex , Ivanov Andrei I.
Published: June 2021

Integrin‐based cell‐matrix adhesions play critical roles in mediating differentiation, survival and motility of epithelial cells. Vesicular trafficking is a key mechanism that regulates assembly of integrin‐based cell‐matrix adhesions and cell migration. A soluble N‐ethylmaleimide sensitive factor (NSF) attachment protein alpha (αSNAP) is a critical regulator of vesicle fusion machinery, however, its role in cell‐matrix adhesions remains unexplored. In this study, we found that siRNA‐mediated knockdown of αSNAP induced matrix detachment in several types of epithelial cells that was independent of a key αSNAP binding partner‐NSF. αSNAP silencing impaired proper glycosylation of β1‐integrin and its trafficking to the cell surface, decreased phosphorylation of FAK and Src and caused disassembly of focal adhesions. Pharmacological inhibition of FAK and Src, but not β1‐ integrin knock‐down recapitulated the effects of αSNAP depletion on cell‐matrix adhesions. Loss of αSNAP depletion resulted in Golgi fragmentation. Furthermore, pharmacological disruption of ER‐Golgi trafficking, or depletion of ER‐Golgi associated protein GBF1 induced FAK/Src dephosphorylation and triggered cell detachment. These results reveal a novel role for αSNAP in modulating β1‐integrin trafficking and matrix adhesions of epithelial cells. Supported by NIH grants DK083968 and DK084953 to AII.
Resolvin D1 Receptor Activation Counter‐regulates H1 histamine receptors in human and rat conjunctival goblet cells
Authors: Hodges Robin R. , Li Dayu , Carozza Richard B. , Jiao Jianwei , Shatos Marie A. , Chiang Nan , Serhan Charles N. , Dartt Darlene A.
Published: June 2021

The purpose of this study was to determine the interactions between H1 histamine receptor (H1R) and GPR32, a GPCR receptor for the proresolution mediator, resolvin D1 (RvD1). Rat conjunctival goblet cells were grown in culture. Chinese hamster ovary (CHO) cells were transfected with human H1R and GPR32 receptor. Intracellular [Ca2+] ([Ca2+]i) was measured in cells loaded with the calcium indicator dye fura 2. In rat goblet cells, the H1R agonist histamine dimaleate significantly increased [Ca2+]i by 327.2 ± 71.5 nM. Exposure to RvD1 significantly inhibited this response to 92.6 ± 21.5 nM. The protein kinase C inhibitor Ro‐ 317549 and an inhibitory peptide of β adrenergic receptor kinase 1 (βARK1) reversed the inhibition of RvD1 on the histamine dimaleate stimulated increase in [Ca2+]i to 197.2 ± 46.3 and 352.7 ± 89.6 nM, respectively. Similar results were obtained in CHO cells transfected with human H1R and human GPR32 receptor. We conclude that RvD1 uses PKC and βARK1 to counter‐regulate the H1R and inhibit its actions.
Neutrophil‐epithelial interactions modulate the inflammatory microenvironment during colitis
Authors: Campbell Eric L , Bruyninckx Walter J , Kelly Caleb , Glover Louise E , Bowers Brittelle E , Bayless Amanda J , Ehrentraut Stefan , Taylor Cormac T , Kominsky Douglas J , Colgan Sean P
Published: June 2021

Neutrophil (PMN) accumulation in crypt abscesses is a pathological hallmark of ulcerative colitis. We hypothesized that transmigrating neutrophils influence the transcriptional profile of intestinal epithelia. Microarray studies revealed a cohort of hypoxia‐responsive genes regulated by PMN‐epithelial crosstalk. Transmigrating PMNs rapidly depleted microenvironmental O2 sufficiently to stabilize epithelial hypoxia‐inducible factor (HIF). Utilizing HIF reporter mice in a TNBS colitis model, we investigated the contribution of PMNs and the oxidative burst to “inflammatory hypoxia”. Gp91phox−/− mice, lacking functional NADPH oxidase, developed exacerbated colitis with enhanced PMN infiltration and diminished inflammatory hypoxia. Pharmacological intervention with a HIF‐stabilizing compound resulted in restoration of mucosal HIF‐signaling concomitant with abrogation of colitis‐severity in the gp91phox−/− mice. In conclusion, transcriptional imprinting of host tissue by infiltrating PMNs modulates host responses to inflammation. Likewise, the oxidative burst contributes fundamentally to localized O2 depletion, resultant microenvironmental hypoxia and effective inflammatory resolution. Funding: CCFA and NIH.
Resolvin E1 enhances phagocytosis‐induced neutrophil apoptosis
Authors: El Kebir Driss , Gjorstrup Per , Filep Janos G.
Published: June 2021

Resolvin E1 (RvE1) is an endogenous lipid mediator derived from the ω‐3 polyunsaturated fatty acid eicosapentaenoic acid that displays potent pro‐resolving actions in experimental models of inflammation. However, little is known about the impact of RvE1 on neutrophil apoptosis, a critical control point in the resolution of inflammation. Culture of human neutrophils with RvE1 did not affect neutrophil longevity, RvE1 accelerated apoptosis evoked by phagocytosis of opsonized Escherichia coli or yeast. RvE1 through the leukotriene B4 receptor BLT1 enhanced ROS generation by NADPH oxidase, leading to subsequent activation of caspase‐8 and caspase‐3. Moreover, RvE1 mitigated phagocytosis‐induced ERK activation. RvE1 also attenuated myeloperoxidase (through Mac‐1), serum amyloid A (through the formyl peptide receptor 2) and bacterial DNA (through TLR9)‐evoked concurrent ERK and phosphatidylinositol 3‐kinase/Akt‐mediated apoptosis‐suppressing signals. These led to reductions in the expression of the antiapoptotic protein Mcl‐1, collapse of mitochondrial transmembrane potential and activation of caspase‐3. Our results identify a novel BLT1 receptor‐mediated mechanism, promotion of phagocytosis‐induced neutrophil apoptosis by which RvE1 could enhance neutrophil apoptosis and subsequently promote inflammation resolution. (Grant support: CIHR MOP‐97742).
Regulation of Mucosal Restitution During Acute Inflammation: JAML Shed From Transmigrating Neutrophils Inhibits Intestinal Epithelial Wound Repair Through Binding to CAR
Authors: Weber Dominique A , Sumagin Ronen , McCall Ingrid C , Leoni Giovanna , Neumann Philipp A. , Andargachew Rakieb , Brazil Jennifer C , Nusrat Asma , Parkos Charles A
Published: June 2021

Neutrophil (PMN) migration across mucosal epithelia is associated with increased permeability, tissue damage and disease symptoms. Previous reports indicate that a junctional adhesion molecule‐like protein (JAML) expressed on PMN binds to an epithelial tight junction (TJ) protein termed coxsackie‐adenovirus receptor (CAR) as one of several steps during PMN transepithelial migration (TEM). To further define the functional relevance of JAML and CAR in PMN interactions with intestinal epithelium, we generated anti‐human JAML mAbs and now show that JAML is shed from PMN during TEM. Similarly, we report stimulus and dose‐related shedding of JAML from activated PMN and monocytes that is dependent on zinc‐metalloproteases. We show that JAML shed from PMN and soluble recombinant JAML inhibit intestinal epithelial barrier function and impair wound healing in a CAR‐dependent manner. Importantly, an anti‐JAML mAb that specifically blocks JAML/CAR binding reversed the deleterious effects of PMN shed JAML on wound closure by increasing epithelial cell proliferation in an ERK‐dependent manner. These findings represent a novel mechanism by which transmigrating myelomonocytic cells actively shed TJ‐binding ligands that alter epithelial barrier function and wound healing, thus contributing to disease pathophysiology.
Modulation of the inflammatory response to enterohemorrhagic E. coli by the secreted mucinase StcE
Authors: Szabady Rose L , Boll Erik J , Welch Rod A , McCormick Beth A
Published: June 2021

Enterohemorrhagic E. coli (EHEC) are human bacterial pathogens that have caused numerous outbreaks of foodborne illness worldwide. EHEC infection causes hemorrhagic colitis that in 5–10% of cases can progress to the potentially fatal hemolytic uremic syndrome (HUS). Although EHEC is not considered a classical inflammatory pathogen, clinical data suggest that inflammation may play a role during intestinal infection and in kidney damage during HUS. Our aim is to understand the molecular mechanisms used by EHEC to elicit a pathologic host immune response. Using our well‐established in vitro model of transepithelial migration of neutrophils, we found that EHEC induce migration of neutrophils across polarized epithelial monolayers. We further demonstrated that secretion of StcE during EHEC infection promotes neutrophil transepithelial migration. StcE is a large zinc metalloprotease, involved in cleavage of the mucin‐type O‐glycosylated proteins CD43 and CD45 at the neutrophil surface. CD43 provides an antiadhesive force in cell‐cell interactions and regulates leukocyte adhesion and migration. Treatment with purified StcE rendered neutrophils more adherent in vitro. Our results suggest that during EHEC infection, modulation of neutrophil migration by StcE may play a pro‐inflammatory role that contributes to a dysregulated immune response and the subsequent development of HUS.
Cellular Dynamics in Salivary Gland Branching Morphogenesis
Authors: Larsen Melinda , Ray Shayoni , Yuan Daniel , Bange Lauren , Dhulekar Nimit , Bhaskaran Abhirami , Otzan Basak , Yener Bulent
Published: June 2021

Branching morphogenesis is a complex developmental process requiring coordination of cell proliferation, cell migration, actomyosin‐mediated cell contractility, and adhesion formation, which is not understood. We utilized computer modeling to simulate cleft progression in the primary bud during salivary gland development. Using computer simulations where individual or multiple parameters were varied in specific subsets of cells and each repeated 100 times, we examined the function of multiple cellular processes in cleft formation. We developed quantitative cleft detection algorithms to report on cleft quality and used confocal imaging of salivary gland organ explant cultures to confirm predictions of the simulations, when possible. Manipulations of the percentage of cells undergoing mitosis and the location of those cells are not currently possible to perform experimentally. Of multiple conditions, computer simulations predicted that a low proliferation rate and equal division of proliferative events between the outer and inner cells in the epithelium produces ideal clefts more than other conditions. This work demonstrates that computer modeling can be used to make predictions regarding cellular mechanisms of branching morphogenesis when experimental manipulation is not technically feasible. Supported by NIH R01DE019244 (M.L. and B.Y.)
Team‐Based Learning: Pedagogy for the 21st Century‐Effective, Efficient and Economical
Authors: Vasan Nagaswami
Published: June 2021

“To teach is to engage students in learning” defines the premise that engaging students in learning is vital to a sustained lifelong commitment to learning. Thus, the teacher role becomes more a designer and facilitator of learning. In medical education problem‐based learning, case‐based learning and concept‐based learning have been adopted with varying degrees of success in last 30 years. There is a growing body of evidence that Team‐Based Learning (TBL) is an effective way to incorporate interactive peer group teaching with enthusiasm for learning. TBL combines various elements of adult learning theories with curriculum model‐ CASE Cooperative, Active, Self‐directed and Experiential Learning in a new and unique way to teach anatomy, which is theoretically multifaceted and content dense. The concept of TBL focuses on learning defined by pre‐class preparation and subsequent in‐class discussion. Faculty organizes the ‘content specific discussion topics’ to help students build baseline facts into a framework of conceptual interpretation and understanding. Millennial student approaches to learning demand teaching that is efficient, engaged, effective, and economical (which ironically agree with the demands of the administration). The presentation will include students’ performances in departmental, NBME shelf and USMLE Step 1 exams as well as survey of student satisfaction in a TBL curriculum.
COORDINATE REGULATION OF CORE EMT REGULTORY FACTORS IS ESSENTIAL FOR MIGRATORY AND INVASIVE BEHAVIOR
Authors: LaBonne Carole
Published: June 2021

A small group of core transcription factors, including Twist, Snail1/2 and Sip1, control epithelial‐mesenchymal transitions (EMTs) and motility during both embryonic development and tumor metastasis. Little is known, however, about how these factors are coordinately regulated in order to mediate the requisite behavioral and fate changes. We have found that a key mechanism for regulating Snail1/2 function in the neural crest is by modulating its stability. We further demonstrated that the stability of Twist and Sip1 is regulated by the same E3 ubiquitin ligase that regulates Snail. This reveals that a common regulatory mechanism has evolved to coordinately control the function of these structurally diverse proteins as they direct migratory and invasive behavior. We further demonstrate that the core EMT regulatory factors physically interact with one another to control each others function, and that their activities are coordinately regulated by GSK3‐beta phosphorylation. Collectively, these findings suggest that the factors that regulate developmental and pathological EMTs/migration must work as a functional unit whose activities are tuned temporally and spatially to ensure proper, coordinated, function. We report a small molecule inhibitor that can intervene in this coordinated regulation, and a novel imaging modality that can be used to follow migratory behavior in opaque tissues.
Longitudinal integrated clerkship training as a model for basic science education
Authors: Hirsh David
Published: June 2021

Longitudinal Integrated Clerkships (LICs), meaningfully different models of medical education, are transforming clinical education worldwide. The success of these innovations begs the question‐‐ how can the LIC design inform the educational structure of basic science training? The LIC model is grounded in educational principles and principles derived from the sciences of learning (general education, medical education, social and cognitive psychology) and thus applicability should be transferable across domains. This presentation reviews the case for change and rationale that drove the wholesale redesign of the clinical year from traditional blocks to an LIC framework. The data about LIC transformation are presented with an eye towards informing possibilities for basic science training. This session is about the evolving anatomy of modern (and classical) educational design.
Anatomy Education Day: A Global Outreach
Authors: Márquez Samuel
Published: June 2021

Education and Outreach forms a powerful two‐pronged approach that provides an effective strategy in addressing the basic educational needs of our high school students (HSS). Explanations for the lack of under represented students in the health care professions range from poor educational opportunities to a lack of exposure to medical institutions. Anatomy Education Day (AED) was sponsored by the AAA and supported by the AMSNY. It brought together the medical schools of New York State to offer aspiring HSS a glimpse into the world of anatomy. SUNY Downstate was the flagship institution that led the AED event on January 23, 2012 affording a hands‐on experience in anatomy that may have sparked interest in pursuing a career in the allied health sciences. HSS were shown human anatomy (heart, brain, lungs, skeleton) and learned about their functions. They presented what they learned to fellow HSS on their return to their schools so that the AED event had a multiplier effect of student exposure. The January 23, 2013 AED was expanded substantially into a worldwide outreach event involving 5 cities in 4 countries simultaneously exposing HSS to anatomy – a subject they may not otherwise have had the opportunity to explore. While the investment of this education outreach program may not bear fruit until years later, history may record the importance of the initial role AED played in assisting to establish diversity in the health sciences.
Development and Diversity of Vascular Smooth Muscle Progenitors
Authors: Majesky Mark William
Published: June 2021

Fate mapping studies show that vascular smooth muscle (VSM) is a mosaic tissue with multiple origins in vertebrate embryos. VSM from diverse embryonic progenitors exhibit distinct gene expression profiles and different functional properties in vivo. Lineage‐dependent pathways for specification of VSM progenitors involve epigenetic reprogramming of the genome guided by position‐specific local environmental cues. Adult vessels also harbor VSM progenitor cells in the media and the adventitia. The origins of adult VSM progenitors are not clear. Adult VSM cells are characterized by plasticity of the differentiated phenotype and they are reported to convert to other mesenchymal cell types (chondrogenic, osteogenic, adipogenic) in vivo. It is possible that intrinsic cellular reprogramming pathways are employed to adopt other cell identities possibly by retracing steps used in embryonic development. If so, VSM progenitors in adult vessel walls may be reflective of such a mechanism.
Vascular extracellular matrix and aortic cell differentiation
Authors: Mecham Robert
Published: June 2021

Vascular extracellular matrix and aortic cell differentiation Robert P. Mecham, Washington University School of Medicine, Department of Cell Biology and Physiology, St. Louis, MO 63110 Beginning with the onset of pulsatile blood flow in the developing aorta, smooth muscle cells in the vessel wall produce a complex extracellular matrix (ECM) that will ultimately define the mechanical properties that are critical for proper function of the adult vascular system. In addition, the vascular ECM provides instructional signals that induce, define, and stabilize vascular cell phenotypes. Hence, the ability to produce ECM is a defining “differentiated” phenotype and the spectrum of ECM molecules that vascular cells produce provides a molecular signature for a specific state of differentiation. Because the formation of a functional ECM must occur in an organized sequence, the “matrix phenotype” is changing throughout the entire period of vessel wall development. ECM production is highest during the late embryonic and early postnatal periods, followed by declining production as blood pressure stabilizes. The adult vascular smooth muscle cell produces little ECM under normal conditions but is capable of activating a repair phenotype in response to tissue injury. Thus, the vascular smooth muscle cell undergoes numerous phenotypic transitions during the formation of the vessel wall that are characterized by the amounts and kinds of ECM molecules produced.
Signaling by the cJun NH2‐terminal kinase
Authors: Davis Roger J
Published: June 2021

The cJun NH2‐terminal kinase (JNK) signaling pathway is implicated in the pathogenesis of diabetes and cancer. High fat diet‐induced obesity causes activation of JNK in target tissues. JNK‐deficient mice are resistant to the effects of feeding a high fat diet, including protection against insulin resistance and failure of obesity development. We have used tissue‐specific JNK‐deficient mice to probe the mechanism of JNK regulation of insulin resistance and obesity. We show that JNK plays different roles in multiple tissues and that the phenotype of whole body JNK‐deficient mice reflects the interactions between these different JNK‐dependent processes. The molecular mechanisms of JNK function in metabolic disease and cancer will be discussed.
The Small Molecule Genistein Increases Hepcidin Expression by Activating Stat3 and Bone Morphogenic Protein Signaling
Authors: Fraenkel Paula G. , Zhen Aileen W. , Nguyen Nancy H. , Gibert Yann , Motola Shmulik , Buckett Peter , Wessling‐Resnick Marianne , Fraenkel Ernest
Published: June 2021

Hepcidin, a peptide hormone produced in the liver, decreases intestinal iron absorption and macrophage iron release via effects on ferroportin. Hepcidin is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. We conducted a small‐scale chemical screen to identify small molecules that modulate Hepcidin expression. We found that treatment of zebrafish embryos with genistein, a major dietary component of soybeans, enhanced hepcidin transcript levels. By treating human hepatocytes (HepG2), we found that genistein caused a significant increase in Hepcidin transcript levels and promoter activity. We discovered that genistein's effect on Hepcidin expression did not depend on estrogen receptor signaling or increased cellular iron uptake, but was impaired by mutation of either the Stat3 binding site or the bone morphogenic protein (BMP) response elements in the Hepcidin promoter. RNA sequencing, chromatinimmunoprecipitation, and ELISA experiments revealed that genistein enhanced BMP and Stat3 signaling in HepG2 cells. In conclusion, genistein is the first nontoxic small molecule identified that increases Hepcidin expression in vivo and in vitro. Genistein and other candidate molecules may subsequently be developed into new therapies for iron overload syndromes. Funding: March of Dimes, NIDDK, US Army Research Office.
Effects of the iron transporter inhibitor ferristatin II on serum iron and lipid metabolism
Authors: Kim Jonghan , Buckett Peter , Wessling‐Resnick Marianne
Published: June 2021

Iron loading is associated with altered lipid metabolism, but underlying mechanisms and clinical consequences remain unknown. We have shown that Belgrade rats, a genetic model of iron‐loading anemia, have elevated serum triglyceride (TG). To explore whether lowering serum iron would influence serum TG, we characterized effects of the small molecule iron transport inhibitor ferristatin II (NSC8679). Rats injected with up to 40 mg ferristatin II/kg twice daily for up to 4 days had lower serum iron and transferrin saturation. Urinary analysis confirmed drug metabolism by acetylation. Rats treated with ferristatin II had lower intestinal uptake of 59Fe, associated with up‐regulation of the iron regulatory hormone hepcidin. Treatment with ferristatin II decreased serum iron in Belgrade rats, and serum TG was significantly reduced. Our study provides an explanation for the relationship between iron loading and lipid metabolism, as well as mechanistic support for interventions that reduce serum iron levels in individuals at risk for hypertriglyceridemia. Supported by NIH DK086774, DK064750 and ES014638.
Fat utilization impacts the negative relationship between insulin sensitivity and Leucine
Authors: Thalacker‐Mercer Anna , Ingram Katherine Heimburger , Garvey W. Timothy
Published: June 2021

The aim was to determine whether identified relationships between serum amino acids (AA) and glucose disposal rate (GDR, assessed w/hyperinsulinemic‐euglycemic clamp) observed in non‐obese (NOI, N=43), obese (OI, N=50), and type 2 diabetic individuals (T2DM, N=31), are influenced by fat utilization (assessed w/respiratory quotient, RQ). Regression analyses revealed that a model with leucine/isoleucine (Leu/Ile) and glycine (Gly) was the strongest predictor of GDR in NOI (R2=0.322, p=0.001), Gly alone was the best predictor in OI (R2=0.276, p<0.001), and Leu/Ile was the only predictor in T2DM (R2=0.234, p=0.011). Partial correlations were performed with cohorts divided by RQ median (Low‐RQ, N=58 and High‐RQ, N=61) to determine whether fat utilization impacted the relationship between GDR and AA. A strong, negative relationship between GDR and Leu/Ile (r=− 0.512, p<0.001) was observed in Low‐RQ subjects (i.e. RQ below the median), while no relationship was detected in High‐RQ. Conversely, a positive association was found between GDR and Gly in both RQ groups (Low‐RQ, r=0.550, p<0.001 and High‐RQ, r=0.304, p=0.021). The results support that among NOI, OI, and T2DM, the availability and/or preference of fat as a fuel source may be a driver of the relationship between Leu/Ile and GDR, but does not impact the relationship between Gly and insulin sensitivity. Support: NIH (DK038765, DK083562, PO1 HL55782), VA Merit
Relationship of cravings with weight loss and hunger: Results from a 6 month worksite weight loss intervention
Authors: Batra Payal J , Das Sai Krupa , Salinardi Taylor , Robinson Lisa , Saltzman Edward , Scott Tammy , Pittas Anastassios , Roberts Susan
Published: June 2021

We examined the association of food cravings with weight loss and eating behaviors in a 6 month behavioral weight loss program conducted in 4 worksites. Assessments included non‐fasting body weight, food cravings (Craving Inventory and Food Craving Questionnaire for state and trait) and eating behavior constructs (Eating Inventory) and were completed by 95 participants. There were statistically significant reductions in all craving variables in the intervention group compared to the controls. Within the intervention group, changes in craving‐trait were significantly associated with weight change after controlling for baseline weight, age, gender and worksite. However, in a multivariate model with craving‐trait and eating behaviors (restraint, disinhibition and hunger), hunger was the only significant predictor of weight change. In contrast to several previous reports of increased food cravings with weight change, this study confirmed a broad reduction in cravings during weight change. In conclusion, reductions in craving‐trait were associated with weight change, but this association not significant when hunger was included in statistical models. Hunger control may directly influence both weight change and food cravings, and studies are needed to examine the effectiveness of hunger suppressing versus craving‐suppressing strategies in behavioral obesity treatment.
Exploring underlying reasons of eating patterns among a low‐income adult population in Lake County, Indiana
Authors: Zhao Yanling , Liu Sandra S. , Mc Carthy Órla M. , McCrory Megan A
Published: June 2021

Background

Evidence suggests that eating patterns, including meal skipping and snacking, may impact health outcomes such as obesity and other chronic diseases. However, why people adopt such eating patterns is poorly understood.

Objectives and Study Design

We conducted one‐on‐one semi‐structured interviews about daily eating and other routines among a convenience sample of 20 employees at a community health center in Lake County, IN.

Results

Participants were aged ≥18 y and mostly African American females. Breakfast was the most commonly skipped meal, followed by lunch and dinner (80%, 60% and 5% of the sample, respectively). In addition, 95% of participants reported ever snacking, and 75% snacked at work or during the day whenever hungry. Only 25% of the sample planned when to eat, and Chi‐square tests indicated this lack of planning may have been related to snacking or skipping breakfast. All participants reported their eating patterns differed on weekends compared to weekdays, with 35% having more time to eat and 30% having different timing of meals on weekends. Overall, being too busy or not having enough time to eat were suggested to be major factors influencing eating patterns.

Conclusion

Not planning an eating schedule may associate with meal skipping and snacking, and hunger may also relate to snacking. Further investigation about how time management influences eating patterns is needed.

Funding: Purdue.
TNFα redistributes ZnT2 to accumulate zinc in lysosomes and activate autophagic cell death in mammary epithelial cells
Authors: Hennigar Stephen R , Lanz Michael C , Kelleher Shannon L
Published: June 2021

Post‐lactational involution coincides with increased levels of inflammatory cytokines such as TNFα in the mammary gland (MG). Injection of TNFα into MGs of lactating mice induces involution; however, the role of TNFα in this process is not well understood. Inflammatory cytokines modulate zinc (Zn) transporters to alter subcellular Zn pools in numerous cell types. Therefore, we hypothesized that TNFα redistributes subcellular Zn pools to activate the autophagic cell death observed in mammary epithelial cells (MECs) during involution. Confocal microscopy showed that MECs (HC11) stimulated with TNFα (15 ng/mL, 18 h) redistributed ZnT2 from the secretory compartment and mitochondria to lysosomes. This coincided with significant increases in lysosomal Zn accumulation (~22%, p < 0.05) and activity of the Zn‐dependent lysosomal enzyme, acid phosphatase (19.2 ± 1.1 vs 25.0 ± 0.8 U/mL; p < 0.05). Zn accumulation led to lysosomal swelling, cathepsin D release, and increased cell death (2.8 ± 0.1 vs 1.1 ± 0.1 × 106 cells/mL; p < 0.05). Importantly, siRNA‐mediated ZnT2 knockdown prior to TNFα‐stimulation abrogated these effects. Collectively, this implicates ZnT2 as a requisite component of TNFα‐induced lysosomal Zn accumulation and activation of autophagic cell death in MECs. Moreover, it suggests that factors augmenting TNFα‐induced Zn redistribution in MECs may lead to premature involution and lactation failure.

Grant Funding Source: NIH R01 HD058614 to SLK
Objectively measured physical activity in overweight and obese pregnant women can be predicted by self‐efficacy and pre‐pregnancy BMI
Authors: Kong Kai Ling , Welch Amy , Campbell Christina , Peterson Anna , Lanningham‐Foster Lorraine
Published: June 2021

Research in understanding the relationship between self‐efficacy (SE) and physical activity participation during pregnancy is limited. Our purpose was to examine the relative contribution of pre‐pregnancy body mass index (BMI) and walking SE in predicting physical activity during pregnancy. Walking SE of 37 overweight and obese pregnant women was assessed using barrier self‐efficacy (BSE) and task self‐efficacy (TSE) questionnaires. Walking was quantified by step counts using a StepWatch™ Activity Monitor. Linear and stepwise regression analyses were used to identify the relative contribution of pre‐pregnancy BMI, BSE and TSE to step count prediction at the end of the second (Tri2) and late third trimester (Tri3). At Tri2, TSE (r2 = 0.254, p < 0.003) and BSE (r2 = 0.123, p < 0.049) significantly predicted step count; however TSE explained 23.4% ( p < 0.003) of the variance, which provided the best model to predict step count according to the stepwise regression analysis. At Tri3, TSE (r2 = 0.234, p < 0.003) and pre‐pregnancy BMI (r2 = 0.167, p < 0.015) significantly predicted step count and both variables explained 36% of the variance (p < 0.001), which provided the best model to predict step count. Interventions designed to increase one's self‐efficacy might be an effective strategy to promote more active lifestyles during pregnancy, especially later in gestation for women with higher pre‐pregnancy BMI.

Grant Funding Source: Iowa State University
Long‐term feeding of dietary slow release glucose reduces daily caloric food intake in vivo
Authors: Yan Like , Philllips Robert , Powley Terry , Kinzig Kim , Kim Choon Young , Hamaker Bruce
Published: June 2021

A true dietary slowly digestible starch is not yet available, thus its long‐term effect on food intake has yet to be elucidated.

Aim

Examine the long‐term effect of a dietary slowly digestible starch material on food intake in vivo.

Methods

Alginate‐entrapped starch microspheres with different slow digesting rates were incorporated into rodent diets. Diet‐induced obese Sprague Dawley rats were assigned to 5 test groups and fed for 8 wks: 1) 14% low‐fat diet; 2) 45% high‐fat (HF) diet; 3) 45% HF with rapidly digestible starch; 4) 45% HF with slowly digestible microspheres; and 5) 45% HF with more slowly digestible microspheres. Meal patterns were monitored using BioDAQ.

Results

A slowly digestible microsphere group had lower daily food intake compared with the 45% HF rapidly digestible starch group (P < 0.05) and similar food intake as the low fat group. Overall meal size was lower in slow digesting microspheres group and meal frequency decreased over time (P < 0.05). mRNA levels of orexigenic NPY and AgRP lowered in microspheres group to a similar level as low‐fat group.

Conclusion

Chronic dietary slow release glucose reduces daily caloric food intake by changing meal size and meal frequency.
Dietary Supplement Use Among Older Adults in the United States, NHANES 2007–2010
Authors: Gahche Jaime Jacqueline , Bailey Regan , Burt Vicki , Dwyer Johanna
Published: June 2021

Sixty‐one percent of older men and 73% of older women reported using at least one dietary supplement (DS) in the past 30 days in the National Health and Nutrition Examination Survey (NHANES) 2007–2010. The purpose of this analysis was to characterize the types of DS products used and the motivations for using them among older adults (ages 60+ yrs; n=4,222) in the NHANES 2007–2010. DSs were categorized by major types (e.g. multi‐vitamin, multi‐minerals (MVMM), botanicals), specific nutrient (e.g. calcium or omega‐3 fatty acids) or intended use (e.g. joint health). The most frequently reported reasons for DS use in older adults were to improve overall health (41%), for bone health (34%), to maintain overall health (31%), for heart health or to lower cholesterol (19%), to supplement the diet (18%), or to prevent health problems (17%). MVMM were the most frequently reported DS (42%) with 46% of MVMM users reporting use to improve overall health and 36% to maintain overall health as the motivations for using them. The second most commonly reported DS was calcium (24%), with 76% of calcium users taking this product for bone health. Omega fatty acids, botanicals, vitamin C, D and E were reported frequently and the motivations for using these products varied. The high usage of DSs in older adults emphasizes the importance of monitoring DS use and understanding the motivations for using them.
A focused ethnographic tool provides insights into socially normative infant and young child feeding (IYCF) practices in the context of a behavior change communications (BCC) intervention in rural Bangladesh
Authors: Zongrone A A , Bhuiyan MI , Singh K , Roopnaraine T , Menon P
Published: June 2021

Social norms influence IYCF practices in all cultures. Challenges remain in studying social norms and tracking them over time. We used an ethnographic tool, free‐listing, to assess socially normative IYCF patterns in the context of a process evaluation of a BCC intervention. Groups of 3–4 women in 90 households in 3 sub‐districts listed all foods usually given to children from 0–24mo and reasons for giving each food. Data on normative age of introduction were analyzed in Stata; text data were analyzed in NVivo. Intervention exposure (IE) was viewing a TV spot, a visit by a frontline health‐worker (FHW), or both. Data analysis captured overall social norms, and by intervention exposure. Under 6mo, breast milk was highly normative (94%), but prelacteals (water (58.9%), sugar‐water (36.7%), honey (43.3%)) were also normative, for ritualistic, cultural and pacifying reasons. Cow's milk and “tinned milk” were normative (76.7%; 1.7 ±2.3 mo & 70.0%; 2.9± 3.6 mo), as was rice gruel (83.3%; 4.7 ± 2.7 mo), and were linked to perceptions of infant need and insufficient breast milk. Fewer households with full IE mentioned breast milk‐substitutes, prelacteals, and cow's milk as early foods. The results highlight that although some BCC recommendations are influencing normative practices, entrenched norms may hamper impact. Support: Alive & Thrive, funded by Bill & Melinda Gates Foundation & managed by FHI360; NIH (5T32HD007331).