bluuebunny/crossref_metadata_2025 huggingface dataset:
The Context‐dependent Effect of Transforming Growth Factor Beta on Valve Interstitial Cell Growth in Vitro
Authors: Li Chen (Maggie) Xi , Rosenthal Alan , Gotlieb Avrum I.
Published: June 2021

Background

Valve interstitial cells (VICs) are the most prevalent cell type in heart valves. We showed that in the early stages of VIC wound repair transforming growth factor beta (TGF‐β) promotes VIC proliferation and repair (A. Liu et al. Am J Pathol 2008; 173:1275–1285). However, the effect of TGF‐β on VIC growth independent of injury is poorly understood.

Materials and Methods

We used normal subconfluent VIC monolayer cultures to test the hypothesis that TGF‐β enhances VIC proliferation. Control and TGF‐β treated VICs were plated at low density. Cell number and proliferation (bromodeoxyuridine) were quantified. Cell morphology was observed and the extent of VIC adhesion to the substratum was quantified using a trypsin adhesion assay.

Results

TGF‐β significantly decreased cell number and altered cell morphology as cells were mostly rhomboid‐shaped as opposed to spindle‐shaped. Cell proliferation was significantly inhibited and treated cells showed significantly more adhesion to the substratum, which may contribute to the observed growth inhibition.

Conclusion

The effect of TGF‐β on VIC growth is context dependent, allowing it to play a dual role by preventing excessive heart valve growth in normal adult conditions and promoting cell proliferation in the early stages of repair, as we reported previously.

Supported by a grant from the Heart and Stroke Foundation of Ontario (grant NA6204).
Association of N‐cadherin and beta‐catenin with activation of valve interstitial cells
Authors: Xu Songyi , Gotlieb Avrum
Published: June 2021

Background

Valve interstitial cells (VICs), the most prevalent cells in the heart valve, become activated upon injury and disease. Cell‐cell adhesion proteins N‐cadherin and β‐catenin should be present in VICs and the latter may translocate to the nucleus upon disruption of cell‐cell contacts to regulate gene expression. We hypothesize that absence or disruption of cell‐cell contacts increases cytosolic β‐catenin and VIC activation.

Methods

Porcine VIC cultures were maintained in standard media in 10% fetal bovine serum for 7 to 10 days. Subconfluent, confluent, and wounded monolayers were fixed and stained with antibodies against β‐catenin and N‐cadherin, and α‐SMA to demonstrate VIC activation. Immunofluorescently stained VICs were viewed under a scanning confocal microscope.

Results

Comparing single, islands, confluent monolayers, and wound edge VICs, we found diffuse, stress fiber and diffuse, reduced, and stress fiber α‐SMA staining respectively. Single VICs and VICs that migrated into the wound showed N‐cadherin and β‐catenin staining in the cytoplasm and lamellapodia with some cells showing nuclear or perinuclear staining for β‐catenin. Cellular islands and confluent monolayers showed co‐localized staining of N‐cadherin and β‐catenin at cell‐cell contacts with weak diffuse cytoplasmic staining.

Conclusion

N‐cadherin and β‐catenin at cell‐cell contacts may regulate VIC activation.

Supported by a grant from the Heart and Stroke Foundation of Ontario (grant NA6204).
Deletion of Lysophosphatidic acid Receptors 1 and 2 Protects Against Lipopolysaccharide‐Induced Acute Lung Injury in Mice
Authors: Zhao Yutong , He Donghong , Pendyala Skrikanth , Berdyshev Evgeny , Goya Jonathan , Chun Jerold , Natarajan Viswanathan
Published: June 2021

Lysophosphatidic acid (LPA), a bioactive phospholipid, exhibits both pro‐ and anti‐inflammatory responses in the airway. LPA activates intracellular signaling pathways by ligating to G‐protein‐coupled LPA receptors (LPA‐Rs). Here, we have evaluated the role of LPA and LPA‐Rs in a LPS‐induced murine model of acute lung injury (ALI). Wild type and genetically engineered LPA1 and LPA2 knockout (LPA1−/− and LPA2−/−) mice were intratracheally challenged with LPS, and after 24 h, bronchoalveolar lavage (BAL) fluids and lung tissues were collected. LPS challenge increased LPA1–3 protein expression in the lung and LPA levels in BAL fluids, as measured by LC‐MS/MS. LPA1−/− and LPA2−/− mice reduced lung inflammation after LPS challenge as determined by IL‐6 levels, and infiltration of neutrophils in BAL fluids. Further, intratracheal administration of Ki16425, an antagonist of LPA1&3, prior to LPS challenge attenuated LPS‐induced IL‐6 release and interstitial infiltration of neutrophils without altering total protein concentration in BAL fluids. Taken together, these results show that inhibition of LPA receptors protects against LPS‐induced lung inflammation, but not pulmonary microvascular injury. These studies identify LPA‐Rs as novel pro‐inflammatory components in LPS‐induced ALI. This work was supported by NIH grant HL 091916 to Y.Z.
CHIP mono‐ubiquitination activity – effects on cardiovascular metabolism and AMPK activity
Authors: Schisler Jonathan C. , Willis Monte S. , Patterson Cam
Published: June 2021

CHIP coordinates protein quality control through its ability to directly ubiquitinate proteins and interaction with cellular chaperones. Mice lacking CHIP expression exhibit exaggerated cardiac hypertrophy in response to one week of pressure overload using transverse aortic banding: a two‐fold increase in heart:body weight and a 30 ± 3% decrease in fractional shortening compared to wild‐type mice. Given the loss of contractile function despite an increase in muscle mass, we further examined the metabolic state of CHIP −/− hearts. After one week of banding, CHIP −/− mice had a 43 ± 10% decrease in mitochondrial biogenesis and 49 ± 2% and 61 ± 7% decreases in steady‐state ATP levels and ex vivo rates of ATP generation, respectively. AMP‐activated protein kinase (AMPK) functions as a cardiac energy sensor, increasing catabolic pathways and mitochondrial biogenesis during times of energy depletion. Surprisingly, there was a 40 ± 4% and 47 ± 6% decrease in the active form and activity of AMPK in banded CHIP −/− hearts, respectively. In cell culture, CHIP is necessary and sufficient for AMPK activation and mono‐ubiquitinates the α, β, and γ subunits of AMPK in vitro. Surprisingly mono‐ubiqutination of AMPKα2 increases kinase activity two‐fold, suggesting for the first time that mono‐ubiquitination can directly regulate kinase activity and demonstrates the vital role of CHIP in cardiac hypertrophy and energy homeostasis.
Ephrin contribution to myelin‐based inhibition of axonal regeneration
Authors: Benson M. Douglas
Published: June 2021

The inability of central nervous system axons to regenerate after traumatic spinal cord injury (SCI) is due, in large part, to the inhibitory effects of myelin. The three major previously identified constituents of this activity (Nogo, myelin‐associated glycoprotein, and oligodendrocyte myelin glycoprotein) were isolated based on their potent inhibition of axon outgrowth, and all transduce their inhibitory signals through the same Nogo receptor/p75 neurotrophin receptor/LINGO‐1 (NgR1/p75/LINGO‐1) complex. In mice, ephrin‐B3 functions during development as a midline repellant for axons of the corticospinal tract (CST), which signal through the axonally expressed EphA4 receptor tyrosine kinase. We demonstrated that ephrin‐B3 is expressed in postnatal myelinating oligodendrocytes and accounts for an inhibitory activity toward EphA4‐bearing axons equivalent to that of the other three myelin‐based inhibitors, acting through p75, combined. Our data were the first description of a known vertebrate axon guidance molecule as a myelin‐based inhibitor of neurite outgrowth. They suggest that susceptibility of a given population of axons to myelin inhibition of regeneration may vary depending on the complement of receptors expressed by that population. Our ongoing studies are determining the extent to which blockage of ephrin‐B3 in vivo can be used to increase regrowth of the functionally critical CST.
Intrinsic control of axon regeneration
Authors: He Zhigang
Published: June 2021

The failure of axons to regenerate is a major obstacle for functional recovery after central nervous system (CNS) injury. Removing extracellular inhibitory molecules results in limited axon regeneration in vivo. To test for the role of intrinsic impediments to axon‐regrowth, we analyzed cell growth control genes using a virus‐assisted in vivo conditional knockout approach. Deletion of PTEN (phosphatase and tensin homolog), a negative regulator of the mammalian target of rapamycin (mTOR) pathway, in adult retinal ganglion cells (RGCs) promotes robust axon regeneration after optic nerve injury. In wild‐type adult mice, the mTOR activity was suppressed and new protein synthesis was impaired in axotomized RGCs, which may contribute to the regeneration failure. Reactivating this pathway by conditional knockout of tuberous sclerosis complex 1 (TSC1), another negative regulator of the mTOR pathway, also leads to axon regeneration. Thus, our results suggest the manipulation of intrinsic growth control pathways as a therapeutic approach to promote axon regeneration following CNS injury.
Novel combination strategies to repair the injured mammalian spinal cord
Authors: Bunge Mary
Published: June 2021

Due to the varied and numerous changes in spinal cord tissue following injury, successful treatment for repair may involve strategies combining neuroprotection (pharmacological prevention of some of the damaging intracellular cascades that lead to secondary tissue loss), axonal regeneration promotion (cell transplantation, genetic engineering to increase growth factors, neutralization of inhibitory factors, reduction in scar formation), and rehabilitation. Our goal has been to find effective combination strategies to improve outcome after injury to the adult rat thoracic spinal cord. Combination interventions tested have been implantation of Schwann cells (SCs) plus neuroprotective agents and growth factors administered in various ways, olfactory ensheathing cell (OEC) implantation, chondroitinase addition, or elevation of cyclic AMP. The most efficacious strategy in our hands for the acute complete transection/SC bridge model, including improvement in locomotion [Basso, Beattie, Bresnahan Scale (BBB)], is the combination of SCs, OECs, and chondroitinase administration (BBB 2.1 vs 6.6, 3 times more myelinated axons in the SC bridge, increased serotonergic axons in the bridge and beyond, and significant correlation between the number of bridge myelinated axons and functional improvement). We found the most successful combination strategy for a subacute spinal cord contusion injury (12.5–mm, 10–g weight, MASCIS impactor) to be SCs and elevation of cyclic AMP (BBB 10.4 vs 15, significant increases in white matter sparing, in myelinated axons in the implant, and in responding reticular formation and red and raphe nuclei, and a significant correlation between the number of serotonergic fibers and improvement in locomotion). Thus, in two injury paradigms, these combination strategies as well as others studied in our laboratory have been found to be more effective than SCs alone and suggest ways in which clinical application may be developed.
hESC‐Derived Definitive Endoderm Induces Cardiomyogenesis in Human Embryonic Stem Cells.
Authors: Van Orman Jordan R , Tayeb Karim Si , Duncan Stephen A , Lough John
Published: June 2021

This laboratory previously reported that chick anterolateral endoderm (AL endoderm) potently induces cardiac myogenesis in mouse embryoid bodies. The requirement to microdissect AL endoderm from gastrulation‐stage embryos precludes the utilization of this tissue to identify cardiomyogenic factors. To overcome these limitations we have assessed whether human definitive endoderm (hDE) cells can mimic AL endoderm. Results indicate that while hDE does not share the cardiomyogenic potency of AL endoderm, both hDE cells and medium conditioned by them significantly enrich cardiac myogenesis in pluripotent hESCs as indicated by rhythmic beating and immunohistochemical/qPCR monitoring of marker gene expression. This response appears to be cardiomyocyte‐specific, since endothelial cell markers are unaffected. The cardiomyogenic effect of hDE is further enhanced when pluripotent hESCs are pre‐induced to mes‐endoderm. These findings demonstrate that, like chick AL endoderm, hDE induces cardiomyogenesis in pluripotent cells, and even more effectively in cells at early stages of mesoderm differentiation (mes‐endoderm). Because this approach is tractable and scalable it should facilitate the identification of novel hDE‐secreted factors, for inclusion in defined cardiomyogenic cocktails to generate homogeneous populations of human cardiomyocytes for use in experimental cell therapy.

Grant Funding Source: NIH
A Guided Electronic Dissector (GED) to Teach Medical Gross Anatomy
Authors: Suarez‐Quian Carlos Andres
Published: June 2021

A Guided Electronic Dissector (GED) was developed to teach medical gross anatomy (Cardiopulmonary [CP], GI and Pelvis/Perineum [PP]) to 180 Special Master's Program (SMP) students integrated into the 1st year medical school class (M1). M1 and SMP attended all lectures and small groups, but only the M1 dissected cadavers as per usual. M1 and SMP were subjected to the same written exams and electronic practicum in CP and GI, whereas in PP the M1 were examined using a cadaveric practicum and the SMP an electronic one. SMP performance in the practicum was no different than the M1 class. Further, the current M1 class performed better when compared to the performance of previous M1 classes in identical questions that were assessed via an electronic practicum versus a cadaveric practicum, respectively. However, the current M1 class performed less well than the SMP when the latter group was assessed using electronic media and the M1 students were assessed using a cadaveric practicum. These results are consistent with the interpretation that anatomy vernacular can be taught and assessed using an electronic approach.
Does quadriceps strength modify the association between patella alta and structural features of osteoarthritis on MRI? The MOST Study
Authors: Stefanik Joshua , Zhu Yanyan , Zumwalt Ann , Gross K. Doug , Clancy Margaret , Lynch John , Frey‐Law Laura , Lewis Cora , Roemer Frank , Guermazi Ali , Powers Christopher , Felson David
Published: June 2021

Patella alta (PA), a high riding patella, is associated with cartilage damage and bone marrow lesions (BMLs) in the lateral patellofemoral joint (PFJ). Quadriceps strength has been shown to decrease cartilage loss in the lateral PFJ. In subjects with PA, the patella is not stabilized by the trochlea; we hypothesized that stronger quadriceps, a dynamic stabilizer, would lead to less PFJ cartilage damage and BMLs, especially in those with PA. We measured the Insall‐Salvati ratio (ISR) [patellar tendon length/patellar length] in 896 knees from a cohort of persons aged 50–79 years. Knees in the highest quartile of ISR were considered to have PA. Quadriceps strength was measured using an isokinetic dynamometer. Lateral PFJ cartilage damage and BMLs were graded on MRI and dichotomized into presence or absence. Using separate logistic regression models, we determined the association between sex specific tertiles of quadriceps strength and cartilage damage and BMLs, among those with and without PA, adjusting for age, sex, and BMI. Among those with PA, subjects in the highest strength tertile were 0.73 (0.4, 1.3) and 0.65 (0.35, 1.2) times less likely to have cartilage damage and BMLs compared to those in the lowest strength tertile. Similar results were seen in subjects without PA. Our results suggest that quadriceps strength may protect against cartilage loss and BMLs but is no more protective for those with than those without PA.

Grant Funding Source: NIH/Arthritis Foundation
Redesigning the Learning Environment for the Millennial Student
Authors: Kvavik Robert B.
Published: June 2021

Redesigning the learning environment for the next generation of students in the health sciences requires space that is flexible, adaptable, and able to support ever‐changing technologies and new learning paradigms. Students benefit from a hands‐on, laboratory‐rich environment where students learn by doing. Students need immediate access to real data and small group workspaces, where IT is highly integrated into all aspects of the learning space.

The presentation outlines ways in which the design of facilities can enable more research and engagement opportunities into the curriculum. Also addressed are success metrics to assess the effectiveness of new learning environments. Expected outcomes are greater ability in problem solving, critical thinking, data analysis and interpretation, laboratory skills, teamwork and communication, and quantitative reasoning.
Melanocyte‐like cells in the heart and pulmonary veins contribute to atrial arrhythmia triggers
Authors: Patel Vickas V , Levin Mark D , Lu Min M , Petrenko Nataliya B , Gupta Tara H , Hawkins Brian J , Buckley Peter T , Yuan Lijun , Beermann Friedrich , Marguiles Kenneth B , Muniswamy Madesh , Eberwine James H , Epstein Jonathan A
Published: June 2021

Atrial fibrillation is the most common clinical cardiac arrhythmia. It is often initiated by ectopic beats arising from the pulmonary veins and atrium, but the source of these beats remains unclear. Here, we report detailed analysis of a cell population within murine and human hearts that displays spatial and cellular characteristics suggesting they contribute to atrial arrhythmia triggers. These cells express genes of the melanocyte lineage, including dopachrome tautomerase (Dct), and populate the pulmonary veins, atria and atrioventricular canal in murine hearts. Murine cardiac melanocyte‐like cells express adrenergic and muscarinic receptors and display transcriptional profiles distinct from dermal melanocytes. Adult mice lacking the gene encoding Dct display increased susceptibility to atrial arrhythmias, despite having structurally normal hearts. Kit mutant mice lacking cardiac melanocyte‐like cells do not develop atrial arrhythmias in the absence of Dct, suggesting that a derangement within these cells, and not their absence promotes arrhythmias. Cardiac melanocyte‐like cells are excitable and those lacking Dct have prolonged repolarization with early afterdepolarizations. Dct‐expressing cells are present in human hearts where atrial arrhythmias can arise. Thus, we propose melanocyte‐like cells in the atrium and pulmonary veins may contribute to atrial arrhythmias in some patients.
Structural Analysis of Heterotrimeric G Proteins and G Protein‐Coupled Receptor Kinases
Authors: Tesmer John J. G.
Published: June 2021

Heterotrimeric G proteins and G protein‐coupled receptor kinases (GRKs) are two relatively small families of enzymes that are turned on via direct interactions with active G protein‐coupled receptors (GPCRs), a large and diverse family of integral membrane proteins. Through these interactions, an amazing variety of environmental cues can induce profound changes in G protein‐regulated processes, such as glucose metabolism and regulation of the actin cytoskeleton, as well as GRK‐regulated processes, such as receptor down‐regulation and MAP kinase activation. Crystal structures of G proteins in complex with their effector targets have revealed many insights into the molecular mechanisms by which heterotrimeric G proteins directly regulate the generation of second messengers (1–4). Structural analysis of GRKs has further demonstrated how heterotrimeric G proteins interact with GRK2 (5, 6), and, most recently, how GRKs are allosterically activated by GPCRs. Based on the available structural data, we propose that heterotrimeric G proteins and GRKs recognize active GPCRs using a similar mechanism.

Support was provided by NIH grants HL071818 and HL086865.
Transcriptional Regulatory Mechanisms in Animal Cells
Authors: Roeder Robert G.
Published: June 2021

Transcriptional regulation by gene‐ and cell‐specific DNA‐binding factors underlies key events in development and in cell growth, differentiation and transformation. Whereas animal cells contain thousands of these factors, their stimulatory or inhibitory effects on the general transcription machinery (RNA polymerases and cognate initiation factors) on specific target genes depend upon complex arrays of cofactors (coactivators and corepressors) that add additional layers of regulation. These cofactors include both chromatin remodeling/histone modifying factors (including various histone acetyltransferases and methyltransferases) and factors (such as the 30‐subunit Mediator complex and the TAF subunits of TFIID) that facilitate more direct communication between promoter‐bound regulatory factors and the general transcription machinery. Most recently, the laboratory has employed biochemically defined cell‐free systems reconstituted with recombinant chromatin templates and purified factors (containing over 90 distinct polypeptides) to identify and to analyze both independent and cooperative functions of diverse types of cofactors, as well as underlying mechanisms.

The function of Mediator, TFIID/TAFs, and selected cell‐specific coactivators and histone modifying factors will be discussed in relation to gene activation by specific transcription factors.
Signal transduction by lipid‐anchored molecules as revealed by single‐molecule tracking
Authors: Kusumi Akihiro
Published: June 2021

Using simultaneous single‐molecule tracking of CD59, glycosylphosphatidylinositol (GPI)‐anchored receptor for controlling self‐attack of the complement complexes, and intracellular lipid‐anchored signaling molecules, Galphai2, Lyn, and PLCgamma, we were able to observe that these intracellular signaling molecules are transiently recruited to ligand‐induced CD59‐cluster rafts, leading to intracellular IP3‐calcium signaling. The key process was the ligand‐induced clustering of CD59, which in turn induces raft‐like nanodomains by concentrating cholesterol and glycosphingolipids. Such an induced raft then recruits Galphai2 and Lyn via both protein‐protein and lipid‐lipid interactions, leading to their greatly enhanced interactions. Such transient recruitment of signaling molecules appeared in various signaling systems we have observed thus far, and could be a general nature of the cellular signaling.
Defining network topologies that can achieve biochemical adaptation
Authors: Tang Chao , Ma Wenzhe , Trusina Ala , El‐Samad Hana , Lim Wendell
Published: June 2021

Many signaling systems show adaptation — the ability to reset themselves after responding to a stimulus. We computationally searched all possible three‐node enzyme network topologies to identify those that could perform adaptation. Only two major core topologies emerge as robust solutions: a negative feedback loop with a buffering node and an incoherent feedforward loop with a proportioner node. Minimal circuits containing these topologies are, within proper regions of parameter space, sufficient to achieve adaptation. More complex circuits that robustly perform adaptation all contain at least one of these topologies at their core. This analysis yields a design table highlighting a finite set of adaptive circuits. Despite the diversity of possible biochemical networks, it may be common to find that only a finite set of core topologies can execute a particular function. These design rules provide a framework for functionally classifying complex natural networks and a manual for engineering networks.
Cooperative phosphorylation in the regulation of Wee1A
Authors: Kim Sun Young , Ferrell James E.
Published: June 2021

The proteins CDK1, Wee1, and Cdc25 collectively form a bistable switch that acts as a trigger for mitosis. One subcircuit of the switch is the double‐negative feedback loop between CDK1 and Wee1; active Wee1 phosphorylates Y15 in CDK1 and thereby inactivates it, and active CDK1 phosphorylates many sites in Wee1 and eventually inactivates it. Here we investigated the mechanism through which the interaction of Wee1 and CDK1 flips from one where Wee1 predominates (in interphase) to one where CDK1 predominates (in M‐phase). We carried out these experiments in Xenopus egg extracts, where Wee1A is the relevant isoform. We found that initially the interaction of Wee1A and CDK1‐cyclin B1 results mainly in inactivation of CDK1, but a small amount of Wee1A phosphorylation also occurs. This Wee1A phosphorylation promotes the interaction of CDK1 with Wee1A, which in turn promotes more Wee1A phosphorylation. This provides a simple, modular mechanism for the cooperative phosphorylation of Wee1A.
Probing and controlling cellular remodeling enzymes
Authors: Wells James A.
Published: June 2021

A group of cellular proteases known as caspases are responsible for a number of fate‐determining steps in higher eukaryotic cell biology including cell death, innate inflammatory responses, cell differentiation, and many other cellular remodeling events. My lab has been developing engineered enzymes to identify the proteins that caspases cleave, and generating new enzymes, small molecules and antibodies to control specific caspase activities in cells. These new tools allow us to dissect the roles these cellular remodelers play, as well to generate new protein and small molecules that may be useful in diagnosing and controlling cancer and inflammation.
New roles for sirtuins in mitochondrial metabolism
Authors: Haigis Marcia
Published: June 2021

Sir2 is an NAD‐dependent deacetylase that connects metabolism with longevity in yeast, flies, and worms. Mammals contain seven homologs of yeast Sir2. Previously we showed that SIRT4, a mammalian Sir2 homolog, is a mitochondrial enzyme that ADP‐ribosylates and down‐regulates glutamate dehydrogenase (GDH), thereby enhancing insulin secretion in the pancreas. Here we show that SIRT4 functions to down‐regulate fatty acid oxidation in the liver. We will discuss implications for SIRT4 function in obesity and age‐related diseases. We will also discuss the coordination of sirtuins in the control of metabolism.
Population genetic inferences from whole‐genome variation data
Authors: Clark Andrew
Published: June 2021

Population genetics allows one to make inferences about many aspects of the past history of an organism, including its past demographic history, mutation patterns, migration history, and the role that natural selection has played on genetic variation. When whole‐genome sequence data are available on multiple individuals within a population, it becomes possible to provide scans across the entire genome for features that relate to these past histories, including genomic regions that are aberrant in having particularly recent or remote common ancestry, or regions that have undergone extensive mutational degradation. Probably the most intriguing application of this approach has been to seek out regions of the genome showing a signature of positive or adaptive evolutionary change. Results of such scans from model organisms will be contrasted with human scans, and highlights will be presented.
Human milk oligosaccharides alter the intestinal epithelial cell surface glycome: A proof‐of‐principle
Authors: Straubinger Kathrin , Ninonuevo Milady , Bode Lars
Published: June 2021

Human Milk Oligosaccharides (HMO) are thought to benefit the breast‐fed infant. However, the exact underlying molecular mechanisms remain unknown. Prebiotic, anti‐adhesive and immunomodulating mechanisms have been postulated. Here, we propose an additional mechanism: Intestinal Glycome‐modification. Pathogen attachment to the intestine is often the initial step of infectious diseases and is mediated by pathogen lectins that bind to the host's intestinal epithelial cell surface glycans. We hypothesize that HMO alter the expression of these cell surface glycans, and thus interfere with host‐microbial interactions. Caco‐2 cell membrane proteins were isolated, N‐glycans released by PNGaseF digest and analyzed by mass spectrometry. Isotopically labeled N‐glycans were used as internal standard. While the relative distribution of high‐mannose N‐glycans was not altered, HMO increased the amount of all individual high‐mannose N‐glycans by 1.8–2.0‐fold. Analysis of Caco‐2 cell mRNA with the glycoV4 oligonucleotide gene array showed that HMO induce differential expression of glycan related genes. Galactooligosaccharides (GOS), currently added to infant formula to mimic HMO, had no effect. In conclusion, our in vitro results establish the proof‐of‐principle that HMO have glycome‐modifying effects. Whether these in vitro results translate to in vivo needs further investigation.
Non‐invasive stool‐based detection of newborn infant gastrointestinal development using gene expression profiles derived from exfoliated epithelial cells
Authors: Donovan Sharon Marie , Davidson Laure A , Zhao Chen , Ivanov Ivan , Goldsby Jennifer S , Lupton Joanne R , Mathai Rose Ann , Monaco Marcia H , Rai Deshanie , Russell Michael , Dougherty Edward R , Chapkin Robert S
Published: June 2021

Understanding of the molecular regulation of intestinal development by human milk has been limited by the lack of non‐invasive approaches. Herein, a novel molecular methodology utilizing stool samples containing intact sloughed epithelial cells was used to identify gene sets (combinations) that were differentially regulated in response to infant feeding. Fecal mRNA was isolated from 3‐month‐old exclusively breast (BF; n=12) and formula (FF; n=10) fed infants. Linear discriminant analysis was used to identify the best single genes and two‐ to three‐gene combinations for distinguishing the feeding groups. EPAS‐1, a transcription factor involved in inducing oxygen‐regulated genes, was identified as a putative “master” regulatory gene using coefficient of determination analysis. Bioinformatic analysis identified differentially regulated processes associated with apoptosis, chemotaxis and development (Wnt and NOTCH pathways). Thus, epithelial cell mRNA isolated from stool has value in terms of characterizing the mechanisms underlying developmentally‐regulated transcriptional activation/repression of genes known to modulate gastrointestinal development. (Funded by Mead Johnson and NIH CA59034, CA129444, DK71707 and P30ES09106)
Multipotent cells from human milk form milk‐secreting alveolar structures in 3‐dimensional culture
Authors: Hartmann Peter Edwin , Thomas Elizabeth , Zeps Nikolajs , Rigby Paul
Published: June 2021

Our objective was to characterize the differentiation potential of cells isolated from human milk in vitro. The total cell population was isolated from expressed human milk samples and cultivated in monolayer culture, then subcultured into 3‐dimensional biomatrices. Differentiation was tracked using the markers CD49f (progenitor), p63 (progenitor), 14‐3‐3σ (Sigma; differentiating epithelial), Cytokeratin 14 (CK14, myoepithelial) and Cytokeratin 18 (CK18; luminal). Expansion of a non‐adherent p63+ population occurred in the first five days of growth, followed by upregulation of Sigma upon adherence in monolayer culture. After 16–21 days of growth, mutually exclusive subsets of the adherent population expressed either CK14 or CK18. In 3‐dimensional biomatrices, single p63+/CD49f+ cells generated organized, multicellular and polarized alveolus‐like spheres with a luminal layer of CK18+ cells and a basal layer of CD49f+ cells that expressed milk proteins in response to prolactin stimulation. This implies that human milk cells provide a novel non‐invasive source of primary cells from lactating tissue to generate in vitro surrogates of mammary secretory epithelium.
Pectin inhibits invasiveness of breast and prostate cancer cells by down‐regulation of urokinase plasminogen activator (uPA) secretion
Authors: Wojnowski Rachael , Jiang Jiahua , Jedinak Andrej , Sliva Daniel
Published: June 2021

Breast and prostate cancers are the most prevalent cancers among women and men in the United States. Pectin is a heteropolysaccharide found mainly in citrus fruits. In the present study, we evaluated the effect of pectin on highly invasive breast [MDA‐MB‐231] and prostate [PC3] cancer cells. We have found that pectin significantly reduced the migration of these cells, however, had no effect on proliferation. Interestingly, when pectin was combined with ProstaCaid & BreastDefend, breast and prostate dietary supplements respectively, it markedly further enhanced inhibition of invasive behavior of breast and prostate cancer cells. These effects were mediated through the down‐regulation of urokinase plasminogen activator (uPA) secretion. Further microarray analysis demonstrated that this treatment affected expression of cell‐cycle regulatory genes e.g. cyclinA2, CDC28, MCM2, TPD1, PCNA and CDKN1A which was confirmed by Western blot analysis. Our data suggests potential use of pectin with ProstaCaid or BreastDefend for the prevention and treatment of breast and prostate cancers, respectively.

Grant Funding Source: Unknown
Meta‐Analysis of Prospective Studies of Red Meat Consumption and Colorectal Cancer
Authors: Alexander Dominik D , Lowe Kimberly A , Cushing Colleen A
Published: June 2021

The relationship between red meat consumption and colorectal cancer (CRC) has been the subject of scientific debate. Thus, to estimate the summary association between red meat intake and CRC and to examine heterogeneity, a meta‐analysis of prospective studies was conducted. Thirty‐four prospective studies of red meat and CRC were identified, of which, 24 represented independent non‐overlapping study populations. Random effects meta‐analysis models were utilized to calculate summary relative risk estimates (SRREs) for high vs. low intake and dose‐response relationships. In the high vs. low intake meta‐analysis of all studies, the SRRE was 1.13 (95% CI: 1.04–1.23), although there was significant heterogeneity (p = 0.016). Summary associations were modified by tumor site and gender. The SRREs for colon cancer and rectal cancer were 1.11 (95% CI: 1.03–1.19) and 1.19 (95% CI: 0.97–1.46), respectively. The SRREs among men and women were 1.21 (95% CI: 1.04–1.42) and 1.01 (95% CI: 0.87–1.17), respectively. The available epidemiologic data are not sufficient to support an independent positive association between red meat intake and CRC. This conclusion is based upon summary associations that are weak in magnitude, heterogeneity between available studies, inconsistent patterns of associations across the sub‐groups analyses, and the likely influence of confounding by other dietary and lifestyle factors.
Effects of trans fatty acid (tFA)‐free fat formulations on lipid and lipoprotein metabolism in gerbils and hamsters
Authors: Kaur Deepinder , Jackson Janice , Oska Angela , Latifi Eno , Khosla Pramod
Published: June 2021

The adverse health effects of dietary tFA resulting from the partial hydrogenation of liquid vegetable oils (PHVO) have prompted health agencies to advocate their reduction or complete elimination from the food supply. Alternatives to PHVO are being considered, including genetically modified oils with reduced PUFA, interesterified fats high in stearic acid, and naturally occurring oil blends with increased SFA. In the current study, we evaluated various oil blends‐ soybean oil (SO), low‐linoleic SO, interesterified SO, coconut oil, a vegetable‐oil blend (palm, corn & sunflower) and a palm oil fraction with an increased content of oleic acid for their effects on lipid and lipoprotein metabolism. In all, 96 male Mongolian gerbils and 75 male Golden Syrian hamsters were fed purified diets for 4 weeks containing ~ 30% calories from the test fat blends and ~0.1% w/w cholesterol. Plasma lipids, lipoprotein concentrations, composition, and particle sizes, and liver free and esterified cholesterol were measured. Despite minor differences between individual diets, only the coconut oil‐based diet (99% coconut oil, 1% SO) induced hypercholesterolemia in both species. These data suggest that within the context of currently recommended prudent diets, various tFA‐free fat formulations can be utilized without compromising plasma lipids. Whether the same occurs in humans is under investigation.
De novo lipogenesis in Type 1 diabetes pre‐ and post‐islet transplant
Authors: Lambert Jennifer Elizabeth , Ryan Edmond A. , Thomson Alan B.R. , Clandinin Michael T.
Published: June 2021

Type 1 diabetes (T1D) patients have elevated cardiovascular disease (CVD) risk yet normal blood lipids, suggesting altered lipid metabolism. Islet transplant (ITx) may cure T1D in some people, but immunosuppressive drugs may be hyperlipidemic. The objective was to investigate lipogenesis in T1D and ITx patients using stable isotopes. People with T1D (T1D; 4F/3M), post‐ITx patients (ITx; 3F/5M) and healthy subjects (Control; 4F/2M) participated. Fasting blood samples (0h and 24h) were analyzed for triglyceride (TG). Deuterium‐labeled water was given and stable isotope analysis performed on VLDL TG estimated hepatic synthesis of total and individual fatty acids (FA). Plasma TG was not different between Control (0.84±0.24), T1D (0.95±0.22), or ITx (0.75±0.31). Compared to Control, T1D showed a trend for greater total FA synthesis, of particularly 14:0, 16:0, and 18:0. ITx had reduced total FA synthesis (p<0.01), particularly 14:0, 16:0, 18:0, and 18:1 (p<0.01 for all) compared to T1D. Total FA synthesis was significantly related to plasma TG in ITx only (p<0.05). Synthesis of 16:0 was correlated with plasma TG level in Control and T1D (p<0.05) but not ITx. T1D may have greater hepatic FA synthesis, contributing to CVD risk. In this ITx group, immunosuppression did not raise plasma TG levels and did not have deleterious effects on hepatic FA synthesis.

Grant Funding Source: Canadian Institutes of Health Research
Differential Oxylipid Metabolism in Response to EPA and DHA in IgA Nephropathy
Authors: Zivkovic Angela M , Yang Jun , Georgi Katrin , Hegedus Christine , Nording Malin L , German Bruce , Hogg Ron J , Weiss Robert H , Hammock Bruce D
Published: June 2021

Immunoglobulin A nephropathy (IgAN) is associated with renal hypertension and inflammation. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in fish oil (FO) improve kidney function in animal models but are inconsistent in humans. Plasma oxylipid profiles from IgAN patients (n=10) and healthy controls (n=12) pre vs. post 2 g/d EPA + 1.5 g/d DHA and IgAN patients pre vs. post corn oil (CO) (n=9) were analyzed by LC/MS/MS. At baseline, patients had significantly higher EPA, DHA, linoleic acid (LA), α‐linolenic acid, and arachidonic acid (ARA) epoxides and diols and higher prostaglandins and leukotrienes. There were isomer‐specific differences between patients and controls pre and post FO. Several EPA and DHA metabolites were increased in both patients and controls post FO, but unchanged in patients post CO. The ARA lipoxygenase metabolites 5‐, 8‐, 9‐, and 15‐HETE and the LA lipoxygenase metabolites 9‐HODE, 9‐oxo‐ODE, and 13‐HODE were decreased in controls but unchanged in patients post FO. There were significant differences between controls and patients pre and post FO supplementation, which are indicative of differences in ω3 and oxylipid metabolism in IgAN.
Improving Nutrition Students' Teaching Self Efficacy Through Leading Cooking Classes in the Community
Authors: Cooke Natalie , Ash Sarah , Carraway‐Stage Virginia , Goodell L. Suzanne
Published: June 2021

Instructors in nutrition need to provide students opportunities to apply academic material and improve teaching skills; service‐learning (SL) is one option. The objective of this study was to determine common challenges among SL students to identify ways to improve their teaching, thereby improving SL outcomes and community programming. Students in this study taught cooking and nutrition classes in the community. Throughout the semester, students critically reflected on their SL experiences. Researchers qualitatively evaluated learnings to determine themes. Students realized that: their personalities impact interpersonal relationships with others; they have unrealistic expectations of peers and community agencies and participants; they are quick to get discouraged when faced with barriers in the community; and they need to be able to adapt their teaching styles to different program participants and improve their communication with community partners and team members. After identifying learnings, instructors can target future instruction to increase students' self‐efficacy and improve the outcomes of SL for the students and the community. SL is an effective way to expose students to the interpersonal communication skills necessary to be a community educator. An NC State Undergraduate Research Grant and Park Scholars' GRASP funded this research.
Online nutrition education for practicing physicians (NEPP)
Authors: Kohlmeier Martin , Adams Kelly , Powell Margo , Zeisel Steven
Published: June 2021

Nutritional interventions can greatly reduce disease burden and mortality throughout the world. Physicians' lack of knowledge and practical skills limits the potential of this simple therapeutic tool. We are generating online instructional modules that give physicians the critical knowledge and skills for nutritional disease prevention and treatment of conditions known to respond to dietary measures.

The presentation is modeled after our widely used Nutrition in Medicine curriculum, using interactions, case studies, and quizzes. The modules describe assessment procedures and therapeutic prescriptions in detail, translating guidelines into actionable practice. The instructional design is tightly focused on assured retention with adaptive remediation. Interactive case studies and skill‐building exercises reinforce the instruction. These interactions serve an additional innovative function of competency assessment by replacing traditional exam questions. Self‐contained, brief units build the instructional paradigm. Designed for busy practitioners, individual modules take only 5–15 minutes to complete. The goal is to prepare learners to immediately use the newly learned skills with their patients.

A constantly growing number of modules is available online (nutritioninmedicine.org) without charge for all physicians in training.

Supported by a grant from the National Cancer Institute.
Non‐Electronic, Non‐Computerized Educational Games for Youth from Music to Nutrition
Authors: Wong Siew Sun , Wong Siew Ngan
Published: June 2021

Objective

Present evaluations of educational game concepts and actual games developed for multiethnic youth aged 4 to 18.

Methods

We applied a set of game concepts to develop music and nutrition educational games. These games were well‐received in classrooms, funfairs, 4‐H and Family Consumer Science conferences. Based on observations and feedback about the most popular games, we created a list of game features that were highly favored by youth. A qualitative pilot study to measure the impact of these games is being conducted in Utah to measure improvement in knowledge, diet, physical activity, and behavior among 8–18 years old youth.

Results

These music or nutrition educational games were tested among >1000 youth in Utah and Malaysia. We found that in general, youth are most attracted to games with sound, next, point increments. Other features that increase engagement are competition yet each player has a fair chance of winning something back, doable tasks that do not require extensive thinking or strenuous physical activity, incentives rewarded by points system, quick turnover, and use of oversize game pieces. One unique feature of these games is that they do not involve screen time but encourage face‐to‐face social interaction, thus balancing the increasingly popular high‐tech learning environment.

Conclusions

These best practices may enrich efforts in educational game development for multiethnic youth.
Amount of food intake is not associated with the liking of the food in preschool‐age children attending a daycare center
Authors: Marshall Ying , Wight A , Bordi Peter , CALDWELL LINDA , KRIS‐ETHERTON PENNY , Kranz Sibylle
Published: June 2021

This study was designed to compare the measured consumption of eight high‐fiber snack foods (cookies, muffins, crackers and cheese, pretzels, dry cereal, applesauce, chocolate pudding, carrots and ranch dip) with the level of liking of the food in children attending a local daycare center. Snacks were served in the mornings of eight different days to a sample of 2–5 year old children (n=42). Food intake was calculated with pre‐and post consumption plate weighing. Liking of the foods was estimated using a 3‐point likert scale (yummy, so‐so, yucky) for the following characteristics: appearance, taste, texture, sweetness, and overall liking; non‐parametric test (p‐value<0.05) was used to determine statistical significance. Results showed that amount consumed and liking of food were not associated except for increased intakes with higher ratings for the appearance and overall liking of the applesauce (p‐value<0.001), the taste of cookies (p‐value<0.04), and the appearance of the ready‐to‐eat cereal (p‐value<0.02). Our results indicate that in these preschool‐age children consuming a morning snack at daycare, food consumption is not predicted by the liking of the food. Further studies to determine the predictors of food intake behavior in this age group are needed.
Sleep duration and risk of obesity among Filipino women
Authors: Borja Judith B. , Adair Linda S.
Published: June 2021

Recent studies have shown a negative association between sleep duration and obesity. We examined this relationship using data on 2002 non‐pregnant women aged 35–68 from the Cebu Longitudinal Health and Nutrition Survey, a community‐based prospective study in Metro Cebu, Philippines. Sleep duration (total hours sleep) was calculated from the women's 24‐hr activity diaries (reported for a typical weekday). Obesity measures include body mass index (BMI; in kg/m2)) and waist circumference (in cm). Linear regression models adjusted for age, energy intake, parity, activity levels, and other confounders show a negative association between sleep duration and BMI [beta (95% CI) −0.26 (−0.38 to −0.14)] and waist circumference[beta (95% CI) −0.57 (−0.88 to −0.27)]. Getting more than 6 hours of sleep protected against the risk of overweight (BMI=>25) [Odds ratio (95% CI) −0.68 (0.55 to 0.84)]. These findings are consistent with those of other studies, and provide valuable insights particularly for older women given that 39% of these women are aged >=50. Among older adults, this sleep‐obesity association gains relevance as sleep duration gets shorter and losing weight becomes more difficult with age.

NIH Grant No. 5R03TW008133‐02
Food Access and Perceptions of the Community and Household Food Environment as Correlates of Fruit and Vegetable Intake among Rural Seniors
Authors: Sharkey Joseph R. , Johnson Cassandra , Dean Wesley R.
Published: June 2021

Little is known about the spatial challenges to good nutrition faced by seniors who reside in rural areas and how spatial access influences fruit and vegetable intake. The purpose of this study is to evaluate the associations among individual and neighborhood characteristics, perceived and objective measures of food access, and fruit and vegetable intake of 589 rural seniors in Texas who participated in the 2006 Brazos Valley Health Assessment. Our analyses revealed that increased distance to the nearest supermarket, food store with a good variety of fresh and processed fruit, or food store with a good variety of fresh and processed vegetables was associated with decreased daily consumption of fruit and vegetables, after controlling for the influence of individual characteristics and perceptions of community and home food resources. Findings suggest that interventions designed to increase fruit and vegetable consumption among rural seniors should consider strategies to ameliorate differential access to healthy food due to food store distance.