bluuebunny/crossref_metadata_2025 huggingface dataset:
O‐GlcNAcylation of STIM1 in pulmonary artery endothelial cells (1176.8)
Authors: Vasauskas Audrey , Patel Payal , Patel Neel , Cioffi Donna
Published: June 2021

Extracellular calcium entry following endoplasmic reticulum store depletion, known as store‐operated calcium entry (SOCE), represents the major Ca2+ signaling event in non‐excitable cells. In the pulmonary endothelium, SOCE through the Ca2+‐selective Isoc leads to increased permeability via inter‐endothelial cell gap formation. Isoc activity may be regulated through post‐translational modification, and indeed, inactivation is phosphorylation‐dependent. Regulation may occur via modification of proteins comprising the channel pore or of accessory proteins such as stromal interacting molecule 1 (STIM1). We showed calcineurin regulates SOCE in pulmonary artery endothelial cells (PAECs) via dephosphorylation of STIM1. In addition to phosphorylation, STIM1 may be regulated by O‐linked β‐N‐acetylglucosamine (O‐GlcNAc), which can have an inverse relationship with phosphorylation on the same residues. Here, we sought to determine whether O‐GlcNAcylation regulates SOC entry in PAECs, and whether this modification has a relationship to phosphorylation. When PAECs were treated with Thaimet G, an inhibitor of O‐GlcNAcylation, SOCE increased. Western blots for O‐GlcNAc showed multiple proteins in PAECs are O‐GlcNAcylated, including a major band at ~85 kDa, likely STIM1. This band decreases upon thapsigargin treatment as phosphorylation increases. These data suggest that O‐GlcNAcylation plays a role and has an inverse relationship with phosphorylation in the regulation of SOCE.

Grant Funding Source: Supported by 1F32HL112565‐01, R00HL089361 and R01HL107778
Misperceptions of statin, metformin, and functional food efficacy by nutrition professionals (118.6)
Authors: Winham Donna , Hutchins Andrea , Thompson Sharon
Published: June 2021

An online survey was sent to 473 Registered Dietitians (74%) and other nutrition professionals (26%) (non‐RDs, health coaches, holistic practitioners, etc.) in Arizona. The purpose was to assess knowledge of the efficacy of functional foods and common medications prescribed to lower cholesterol and control postprandial hyperglycemia in persons with T2DM. Respondents were asked to rank: statins, dried beans, oatmeal, soy protein, and red wine for their effectiveness in lowering LDL cholesterol. Only 65% ranked statins first and 47% ranked oatmeal second. Beans are clinically more effective than oatmeal but were ranked 3rd by 38% with red wine and soy ranking 4th and 5th. Significant differences were observed by training category with RDs having more accurate knowledge than non‐RD nutrition professionals. A similar pattern was observed with responses for T2DM treatments. Overall, 71% ranked metformin 1st, with beans ranked 2nd, cinnamon 3rd, vinegar 4th, and red wine in 5th place for efficacy. Significant differences by professional category were seen for metformin (57% Non‐RDs vs. 76% RDs), beans (31% Non‐RDs vs. 50% RDs), and cinnamon (28% Non‐RDs vs. 38% RDs). Knowledge differences by professional training are not surprising, but it is cause for concern when only 71% and 76% of RDs think statins and metformin are the most effective options. RDs were more knowledgeable than non‐RDs, but large gaps and misperceptions about functional foods vs. medications for controlling hypercholesterolemia and postprandial hyperglycemia warrant greater outreach and education efforts by nutrition educators for clinicians as well as consumers.
Ephrin‐A1 regulates cell dynamics through molecular activation, mechanical tension, and morphology (1180.13)
Authors: Hur Sung Sik , Yeh Yi‐Ting , Vuong Edward , Min Peter , Nguyen Phu , Wang Kuei‐Chun , Li Yi‐Shuan , Wang Yingxiao , Chien Shu
Published: June 2021

Ephrins, the Eph family of receptor tyrosine kinases and their ligands, have been shown to be key players in guiding cell migration, polarization, and cell‐matrix and cell‐cell adhesions. Specifically, the ligand Ephrin‐A1 has been shown to regulate cell motility and morphology through the activation of EphA receptors, which signal to PI3K pathway to induce cell contraction. We performed live cell experiments on mouse embryonic fibroblasts (MEFs) to study the interrelations of molecular activation of myosin light‐chain kinase (MLCK) as determined by fluorescence resonance energy transfer (FRET), mechanical tension as measured by using novel 3D intracellular force microscopy (3D IFM), and cell morphology in response to Ephrin‐A1. The results demonstrate positive correlations among the elevation of MLCK, an increase in intracellular mechanical tension, and a reduction of cell area, which indicates an augmentation of contraction. These effects of Ephrin‐A1 were absent in MEFs with PI3K subunit p85b knockout (p85b‐/‐), indicating that p85b in PI3K plays a significant role in mediating the chemical, mechanical, and morphological effects of Ephrine‐A1. This study serves to elucidate the mechano‐chemical regulation of cell dynamics under Ephrine‐A1 guidance.

Grant Funding Source: Supported by NIH HL108375 (Shu Chien), NSF: (Shaochen Chen)
TNFα regulates endothelial progenitor cell migration through the NF‐κB pathway (1180.16)
Authors: Prisco Anthony , Hoffmann Brian , Kaczorowski Catherine , Greene Andrew
Published: June 2021

Many therapies exist to treat myocardial infarctions (MI). Most current therapies focus on management of symptoms rather than regeneration of tissue. Cardiac function has been improved in rodents using bone marrow derived endothelial progenitor cells (EPCs) as a regenerative therapeutic, however human studies have produced inconsistent results as regenerative mechanisms are poorly understood. For an injected EPC to improve cardiac function, the cell must migrate, bind and engraft within the damaged tissue. EPCs are mobilized from the bone marrow to the circulation following MI. One of the inflammatory cytokines released in an MI is TNFα. We hypothesized that an acute dose of TNFα would induce EPC migration toward vessel like structures in vitro. To test this hypothesis, endothelial cells were plated onto a 4 well chamber slide coated with Matrigel substrate to grow vessel‐like structures. After 48 hours, EPCs were treated with TNFα or vehicle and added to the wells containing vessel‐like structures. After tracking the EPCs for 16 hours, a significantly higher fraction of EPCs treated with TNFα than vehicle were found to have migrated towards vessel like structures. Follow up analysis demonstrated that the migratory phenotype was mediated through the NF‐κB pathway, simulated by the TNF Receptor 2. Proteomic analysis suggested several NF‐κB transcriptional targets that were directly tested using siRNA knockdown.

Grant Funding Source: NIH Grant HL082798
A method for bridging cell‐based transcriptomic and systematic data in obesity and aging studies (1180.2)
Authors: Tan Ningzhi , Zhou Yanjiao , Gao Ling , Yao Yisang , Dong Yunzi , Lu jihong , Zhu Jia‐Shi
Published: June 2021

We have reported the challenges of directly interpreting transcriptomic data at systems physiology levels, when hyper‐/hypotrophy or hyper‐/hypoplasia occurs (FASEB J 2007, 21:A1352). We found (1) some gene expressions were under whole body physiological controls; (2) some reflected compensatory feedback control or decompensation at organ levels. Transgenic manipulations on a gene or genes often follow the transcriptomic studies, but may lead to non‐physiological phenotypes and non‐pathophysiological interruption of the gene regulations and the cascade of signal transduction. Here we report a method to bridge the cellular transcriptomic data and tissue/systems physiology. In obesity studies with cell hypertrophy and hyperplasia and enlarged liver and adipose tissue masses, the collected tissues were weighted in whole and in small divided pieces before freezing immediately for DNA, protein and activity tests or after being socked in RANlater for gene expression test. The total DNA and RNA concentrations were measured as the indicators of cell number, assuming that total DNA is constant per cell at normal or disease conditions. Such middle information allowed us to convert the cellular transcriptomic data under the conditions with changes in cell size and number and tissue mass to a tissue/organ level and to interpret the outcomes under the regulations of systems physiology and pathology.
Holistic multilevel modeling of type‐2 diabetes (1180.5)
Authors: de Graaf Albert , van Ommen Ben
Published: June 2021

Type‐2 diabetes (T2D), is presenting a rapidly growing health and economic problem. For personalized lifestyle/medical treatment advice, it is essential that computer modelling tools become available to integrate the results from diagnostic tools that probe the function of all involved organs/processes in a single holistic view. Our objective is to build such a model for T2D, an important comorbidity of metabolic syndrome.

As a first step towards fully personalized quantitative diagnosis and prediction we developed a descriptive model based on causal loop diagrams at a very high aggregation level that integrates mechanisms across all relevant domains including body weight dynamics, glucose/insulin dynamics, inflammation, gut health, and mental stress. The model integrates qualitative and semi‐quantitative information and expert knowledge.

The model was able to simulate the development of T2D over a multi‐year period following different food intake profiles, as well as disease modulation by increased physical activity or mental stress‐relieving lifestyles.

In a recently started EU project MISSION‐T2D (www.mission‐t2d.eu), the high aggregation level model will be interlinked with a model at a second aggregation level that describes the integrated dynamics of all processes at the minute‐day timescale. This allows to enrich the model with mechanisms probed by PK‐PD studies and nutritional challenge tests and should pave the way for translating validated multilevel immune‐metabolic models into the clinical setting of T2D.

Concluding, we have successfully made a first step towards a multiorgan, multilevel systems health model of T2D that can be used to integrate diagnostic data from different domains for personalized lifestyle/treatment advice.
Cholesterol inhibits ROMK channels by holding PI(4,5)P2 in microvilli (1181.17)
Authors: Liu Bing‐Chen , Yang Li‐Li , Lu Xiao‐Yu , Song Xiang , Li Xue‐Chen , Chen Guangping , Eaton Douglas , Shen Bao‐Zhong , Ma He‐Ping
Published: June 2021

The kidney plays an important role in maintaining ion balance in the blood. K+ balance is tightly regulated by a K+ channel called ROMK1. We used cultured mouse cortical collecting duct principal cells (mpkCCDc14 line) to determine how ROMK1 is regulated by cholesterol. Previous studies suggest that cholesterol inhibits ROMK channels via a direct mechanism. Surprisingly, our data demonstrate that ROMK1 channels are not located in cholesterol‐rich lipid rafts in cultured CCD cells. Patch‐clamp data demonstrated that a small‐conductance K+ (ROMK1) channel in mpkCCDc14 cells was inhibited by cholesterol and activated by extraction of cholesterol with methyl‐beta‐cyclodextrin (MbetaCD). However, confocal microscopy data showed that ROMK1 was not in the microvilli where cholesterol‐rich lipid rafts are located, but in the planar region of the apical membrane of mpkCCDc14 cells, and that phosphatidylinositol‐4,5‐bisphosphate (PI(4,5)P2), a well‐known activator of ROMK channels, was found only in the microvilli under resting conditions. Interestingly, after cholesterol synthesis was inhibited by lovastatin, PI(4,5)P2 diffused into planar region with a consequence of activation of ROMK channels. Activation of ROMK1 channels by MbetaCD was reversed by addition of exogenous cholesterol, but the activation did not occur when PI(4,5)P2 was sequestered with its antibody. These results suggest that cholesterol inhibits ROMK1 channels, at least in part, by limiting PI(4,5)P2 diffusion from microvilli to planar region of the renal CCD cell apical membrane. Since we have reported that cyclosporine A (CsA), an immunosuppressant drug which causes hyperkalemia, can alter the levels of cholesterol in renal epithelial cells, this study provides a basis for researchers to determine the mechanism of CsA‐induced hyperkalemia.

Grant Funding Source: supported by NIH 5R01‐DK067110 to HPM and by NSF of China 81130028 to B‐ZS
Anchored PDE4 controls CFTR conductance in normal and cystic fibrosis airway epithelia (1181.3)
Authors: Richter Wito , Blanchard Elise , Zlock Lorna , Lao Anna , Mika Delphine , Namkung Wan , Xie Moses , Scheitrum Colleen , Gruenert Dieter , Verkman Alan , Finkbeiner Walter , Conti Marco
Published: June 2021

Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) that impair its expression or function. PKA phosphorylation is the main mechanism to activate CFTR. However, previous reports indicated that cAMP/PKA activators are ineffective in restoring CFTR conductance in CF epithelia. Our current findings challenge this conclusion. We show that inhibition of type 4 cAMP‐phosphodiesterases (PDE4s) increased CFTR activity in non‐CF primary human bronchial epithelial cells under basal conditions and after β‐adrenergic stimulation, and slowed the return of stimulated CFTR activity to basal levels. In cells homozygous for ∆F508‐CFTR, the most common mutation found in CF patients, PDE4 inhibition alone produced minimal channel activation. However, PDE4 inhibition strongly amplified the effects of CFTR correctors, drugs that increase expression and membrane‐localization of CFTR, and CFTR potentiators, drugs that increase channel gating to reach ~25% of the chloride conductance observed in non‐CF cells. Biochemical studies indicate that PDE4s are anchored to CFTR and mediate local regulation of channel function. Taken together, our results implicate PDE4 as an important determinant of CFTR activity in airway epithelia and support the use of PDE4 inhibitors to potentiate the therapeutic benefits of CFTR correctors and potentiators.

Grant Funding Source: Supported by the Cystic Fibrosis Foundation, the NIH, and Cystic Fibrosis Research Inc.
β1‐Adrenergic receptor signaling activates TRPV5 via the protein kinase A pathway (1181.4)
Authors: Hoenderop Joost , van der Hagen Eline , Tudpor Kukiat , Verkaart Sjoerd , Bindels Rene
Published: June 2021

Epinephrine (Epi) and norepinephrine (NE) are present in the pro‐urine. Active Ca2+ reabsorption in the late distal convoluted and connecting tubules (DCT2/CNT) is initiated by Ca2+ influx through the transient receptor potential vanilloid type 5 (TRPV5) Ca2+ channel. Although it was reported that β‐adrenergic receptors (β‐AR) are present in the DCT2/CNT region, their role in active Ca2+ reabsorption remains elusive. Here we revealed that β1‐AR, but not β2‐AR is colocalized with TRPV5 in DCT2/CNT. Subsequently, treatment of TRPV5‐expressing mouse DCT2/CNT primary cell cultures with the β1‐AR agonist dobutamine showed enhanced apical‐to‐basolateral transepithelial Ca2+ reabsorption. In human embryonic kidney (HEK293) cells, dobutamine was shown to stimulate cAMP production, signifying functional β1‐AR‐expression. Fura‐2 experiments demonstrated increased activity of the TRPV5 Ca2+ channel with dobutamine, which could be prevented by the PKA inhibitor H89. Moreover, non‐phosphorylable T709A‐TRPV5 and phosphorylation‐mimicking T709D‐TRPV5 mutants are unresponsive to dobutamine. Surface biotinylation showed that dobutamine did not affect plasma membrane abundance of TRPV5. In conclusion, activation of β1‐AR stimulates active Ca2+ reabsorption in DCT2/CNT, increase in TRPV5 activity via PKA phosphorylation of residue T709 possibly plays an important role.
Absence of Slc26a9 results in altered tracheobronchial anion transport and high mortality in neonate mice (1181.7)
Authors: Liu Xuemei , Riederer Brigitte , Anagnostopoulou Pinelopi , Li Taolang , Rausch Brigitte , Duerr Julia , Soleimani Manoocher , Mall Marcus , Seidler Ursula
Published: June 2021

Background:The anion transporter Slc26a9 is highly expressed in lung and stomach. We observed an excess perinatal mortality in Slc26a9‐/‐ neonate mice. Aim and Methods: In search for the underlying cause, the dead pups were collected, genotyped, and examined histologically. Slc26a9 and CFTR mRNA expression was measured by qPCR in microdissected lung. Slc26a9‐/‐, CFTR‐/‐, and WT neonatal trachea were studied electrophysiologically in Ussing chambers. Results: Dead pups were Slc26a9‐/‐ in >95%. Volume density measurements revealed a strong excess of tracheal and bronchial mucus in the Slc26a9‐/‐ pups. Slc26a9 as well as CFTR mRNA expression was significantly higher in the distal than proximal airways, and significantly higher in neonatal than adult mice. The amiloride‐insensitive, bumetanide‐sensitive short circuit current (Isc) was higher in neonatal Slc26a9‐/‐ than WT trachea, but lower in neonatal CFTR‐/‐ than WT trachea. Surprisingly, forskolin (FSK) + IBMX elicited a significantly higher Isc response (∆Isc) in neonatal Slc26a9‐/‐ than WT trachea. This difference was abolished by preincubation with the CFTR inhibitor CFTRinh172. Conclusions: Neonatal Slc26a9‐deficient trachea displayed a higher electrogenic anion secretory rate than WT, and this difference was abolished by CFTR inhibition. The early death of Slc26a9‐/‐ pups may be related to problems with airway liquid absorption rather than lack of secretion.

Grant Funding Source: Supported by DFG Se 13‐4 and 9‐6 to Ursula Seidler
Alkaline pH regulates the epithelial sodium channel (1181.8)
Authors: Ochoa Luis , Peterson Zerubbabel , Collier Daniel , Snyder Peter
Published: June 2021

The extracellular domain of the epithelial Na+ channel (ENaC) is exposed to highly varied environments, and has been postulated to function as a sensor that modulates channel activity in response to changes in the extracellular environment. In the kidney collecting duct, pH varies widely (4.5‐8.5). We previously found that acidic pH reversibly stimulates ENaC whereas alkaline pH reversibly inhibits ENaC. Eight extracellular domain residues are required for regulation by acidic pH, but mutation of these residues did not alter inhibition by alkaline pH, indicating that alkaline pH regulates ENaC by a different mechanism. Here we tested the effect of extracellular alkaline pH on ENaC current by expressing human ENaC in Xenopus oocytes and recording ENaC Na+ currents as the extracellular pH was altered. We identified the basic residue Arg‐90 in the extracellular domain of γENaC that is in part responsible for ENaC inhibition by alkaline pH. Mutation of this residue to neutral residues dramatically reduced inhibition. When the positive charge was reintroduced by MTSET modification of Cys at this position, the response to alkaline pH was partially restored. Based on homology modeling, we predicted that Arg‐90 is located in close proximity to Glu‐446 in βENaC. Consistent with this prediction, crosslinking of cysteines introduced simultaneously at the two positions (γR90C and βE446C) with MTS‐14‐MTS reduced ENaC current and Na+ self‐inhibition. Together, the data support a role for Arg‐90 in ENaC regulation by alkaline pH. We speculate that titration of this residue induces intersubunit conformational changes that modulate ENaC gating.

Grant Funding Source: NIH
Astrocytic localization of supraoptic serine racemase and its potential participation in NMDA‐stimulated vasopressin responses in hypothalamo‐neurohypophysial explants (1182.9)
Authors: Valdez Joe , Valdez Matt , Kim Simon , González Gwendolyn , Fernández Ruben , Gutiérrez Robert , Currás‐Collazo Margarita
Published: June 2021

Our lab has previously shown that vasopressinergic magnocellular neuroendocrine cells (MNCs) express various NMDA receptor subunits including the obligatory NR1. Other reports show that NMDA receptors (NMDAr) mediate osmotically‐induced vasopressin (VP) release from the hypothalamo‐neurohypophysial system (HNS). Recently, Panatier et al 2006 revealed that D‐serine is the endogenous coagonist for NMDAr in MNCs of the supraoptic nucleus (SON). Therefore, we investigated the role of D‐serine in NMDAr‐mediated VP release from acutely dissected HNS explants taken from adult male C57BL/6 mice and incubated in a perifusion chamber from which samples were analyzed for vasopressin (VP) using EIA (Gregg & Sladek 1984). After a 4 hr incubation, explants were treated with 5‐100 uM NMDA for 1 hr in the presence and absence of SR inhibitor, L‐Aspartic acid β‐hydroxamate (LA‐βH; 3 mM). We found that NMDAr‐mediated VP release was reduced by LA‐βH suggesting that SR activity and D‐serine signaling are required. In immunohistochemical experiments using adult SON sections, we found marked colocalization of green SR immunoreactivity (ir) with either of two astrocyte specific markers, glial fibrillary acidic protein (GFAP) and glial excitatory amino acid transporter GLT‐1 labeled red. In other experiments animals were injected with Evans Blue (EB), which retrogradely labels VP and oxytocin‐producing MNCs projecting to the posterior pituitary with red fluorescence (excitation/emission maxima ~540/680 nm). SR ir was examined under two osmotic states: hyperosmotic (3.5M NaCl) or normosmotic (0.9% NaCl; 0.6cc/100g bw, i.p.). EB‐positive MNCs showed little or no SR ir, which was redistributed in hyperosmotic animals. We hypothesize that a significant proportion of SR is expressed in astrocytes and that D‐serine gliotransmission contributes to VP responses in SON MNCs.

Grant Funding Source: Supported by APS STRIDE summer fellowship (JV), APS Porter fellowship (MV), and MARC (GG, RF, RG).
Middle cerebral artery mean flow velocity changes to non‐pharmacologically induced hypertension and hypotension in humans (1184.6)
Authors: Brassard Patrice , Paquette Myriam , Le Blanc Olivier , Gaudreau Alexandra , Moreau Pascale‐Renée , Clément Andrée‐Anne , Thibault Guy
Published: June 2021

Asymmetrical responses of cerebral blood flow in response to elevating vs. falling blood pressure has been reported using cyclic inflation and deflation of thigh cuffs in head injury patients and pharmacologically in healthy volunteers. Whether this phenomenon exists with non‐pharmacologically induced hypertension and hypotension is unknown. In this study, we examined the relative changes in cerebrovascular conductance index (CVCi), defined as percent changes in middle cerebral artery mean flow velocity / percent changes in mean arterial pressure, during transient hypertension vs. hypotension in 10 healthy male volunteers {age [median (range)]: 24 (22‐35 yrs), body weight [mean±SD: 72±12 kg]; maximal oxygen uptake: 55±5 ml/kg/min}. Transient hypertension and hypotension were induced via squat‐stand manoeuvres executed at 0.025, 0.05 and 0.1 Hz. The CVCi responses to hypertension (squat) compared to hypotension (stand) were attenuated at 0.05 Hz (0.57±0.38 vs. 1.84±0.56 %/%) and 0.1 Hz [0.83±0.41 vs. 1.59±0.57 %/%; all p<0.01), but not at 0.025 Hz [0.86±0.30 vs. 1.12±0.31 %/%; p=0.1). These results suggest that the human cerebrovascular function is better adapted to combat non‐pharmacologically and physiologically relevant transient hypertension than transient hypotension.

Grant Funding Source: Supported by a research contract from the Ministère de l’Éducation, du Loisir et du Sport (Québec)
Induction of autophagy by zinc during acute ethanol exposure (122.1)
Authors: Liuzzi Juan , Menzel Lorenzo , Yoo Changwon
Published: June 2021

Autophagy is an important catabolic process for the clearance of damaged or superfluous proteins and organelles. The role of ethanol in the regulation of autophagy remains controversial, with some studies showing induction of autophagy by ethanol while others show inhibition. Nevertheless, the view that maintenance of adequate levels of autophagy protects against ethanol induced damage is widely supported. We investigated the role of zinc in the modulation of autophagic flux in human hepatoma cells VL‐17A. LC3II/LC3I ratio, p62 and Beclin‐1 expression and autophagosomes number were analyzed in cells incubated in medium containing 0, 10, 20 or 40µM of zinc with or without 100 mM of ethanol, in presence or absence of the autophagy inhibitor chloroquine. The effect on autophagic flux of co‐incubation with lipopolysaccharide was also investigated. Lastly, labile zinc was analyzed in cells exposed to ethanol and the autophagy inhibitor 3‐methyladenine. Excess zinc had an additive effect on the induction of autophagy by ethanol and lipopolysaccharide. In addition, 3‐methyladenine treatment decreased labile zinc in cells. Our results indicate that zinc is a positive regulator of autophagic flux and support the notion that zinc protective effects against ethanol induced damage are mediated by autophagy.

Grant Funding Source: Supported by NIH 1R03AA022451‐01
Associations between tissue retinol binding protein‐4, visfatin and vaspin protein concentrations and blood triglyceride and cholesterol levels in morbidly obese subjects (133.3)
Authors: Goktas Zeynep , Wang Shu , Besler Tanju
Published: June 2021

Background: Obesity is associated with insulin resistance, diabetes, and cardiovascular diseases. Retinol binding protein‐4 (RBP‐4), visfatin and vaspin are known to have some effect on insulin resistance although their effects on cardiovascular diseases are not clear. Our research focuses on tissue secretions of RBP‐4, visfatin and vaspin proteins and their relationships to blood lipids.

Methods: Thirty eight morbidly obese (BMI 蠅40) subjects had the subcutaneous, omental, and mesenteric adipose, liver and muscle tissue samples taken at the time of Roux‐n‐Y surgeries. Total proteins were extracted with RIPA solution and measured with ELISA kits (Adipogen, Korea). Blood triglycerides and cholesterols levels were measured using standard methods. Results were analyzed using Pearson correlation with p <0.05 being significant.

Results: Omental adipose tissue visfatin protein concentrations were positively correlated with blood total cholesterol levels (r = 0.382, p = 0.018). Muscle tissue visfatin protein concentrations were negatively correlated with blood HDL cholesterol levels (r = ‐0.384, p = 0.017). Omental adipose tissue RBP‐4 protein concentrations were positively correlated with blood triglyceride levels (r = 0.404, p = 0.012). Mesenteric adipose tissue vaspin concentrations were positively correlated with blood total cholesterol levels (r = 0.429, p = 0.007).

Conclusions: Our findings indicate that tissue RBP‐4, visfatin and vaspin protein concentrations might have effects related to dyslipidemia during obesity.
Lipocalin2 regulates the p38MAPK‐PGC1α‐UCP1 pathway to activate the thermogenic program in subcutaneous white adipocytes via a non‐adrenergic mechanism (133.5)
Authors: Deis Jessica , Guo Hong , Chen Xiaoli
Published: June 2021

Thermogenic activity in adipocytes is negatively correlated with obesity. Adrenergic activation of p38 mitogen‐activated protein kinase (p38MAPK) mediated thermogenesis is well established. Recently, non‐adrenergic regulation of thermogenesis has gained interest. Lipocalin 2 (Lcn2) is a novel adipokine whose deficiency induces cold intolerance in mice. Herein, we explored the thermogenic effect of Lcn2 and its underlying mechanism in subcutaneous white adipocytes. Inguinal stromal‐vascular cells were collected from wild‐type (WT) and Lcn2‐/‐ mice and differentiated to adipocytes. Lcn2‐/‐ mice had a downregulation of uncoupling protein 1 (UCP1) expression in inguinal adipose tissue (iWAT) compared to WT mice on a high fat diet. Lcn2‐/‐ inguinal adipocytes showed significantly lower UCP1 and peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC1α) expression as well as p38MAPK phosphorylation when compared to WT cells. However, norepinephrine‐induced UCP1 and PGC1α transcription, lipolysis and p38MAPK phosphorylation remained unchanged in Lcn2‐/‐ adipocytes. Additionally, Lcn2 recombinant markedly upregulates UCP1 and PGC1α transcription, and increases p38MAPK phosphorylation in Lcn2‐/‐ inguinal adipocytes. This effect is more pronounced under insulin‐stimulated conditions, suggesting Lcn2 synergizes with insulin to activate thermogenic programming. Moreover, a p38MAPK inhibitor attenuated Lcn2‐induced UCP1 expression. In conclusion, Lcn2 has a significant role in promoting iWAT thermogenesis by upregulating the p38MAPK‐PGC1α‐UCP1 pathway via a non‐adrenergic mechanism.

Grant Funding Source: Supported by NIDDK
Adipogenesis and glycemic improvement can be concurrent, yet independent (133.7)
Authors: Hegde Vijay , Dubuisson Olga , Dhurandhar Nikhil
Published: June 2021

Although excessive adiposity is linked with impaired glycemic control, in some animal models transgenic manipulations that increase adiposity, paradoxically improve glycemic control. In mice, Ad36, ‐a human adenovirus, promotes adipogenesis, and enhances glycemic control. The E4orf1 gene of Ad36 is sufficient to improve diet induced hyperglycemia in mice, which offers therapeutic potential. Studies indicate that via E4orf1, Ad36 increases Ras to up‐regulate PPARγ and Glut4 signaling, leading to adipogenesis and better glycemic control, respectively. Our objective was to determine if the adipogenic and glycemic signaling pathways induced by E4orf1 are interdependent. PPARγ null mouse embryonic fibroblasts were transfected with vector expressing E4orf1 or a null vector. Compared to cells transfected with the null vector, E4orf1 expression significantly increased glucose uptake despite a complete absence of PPARγ or adipogenesis, and significantly enhanced Ras activation and the protein abundance of glucose transporters Glut4, Glut1 and adiponectin. Thus, E4orf1 can enhance the signaling underlying glucose disposal, independent of adipogenic signaling. This offers the possibility to harness the anti‐diabetic property of E4orf1, without its adipogenic effect. Moreover, conceptually, these results suggest that adipogenesis and glycemic improvement can be concurrent, yet independent.

Grant Funding Source: Vital Health Interventions
Designing, developing, and implementing effective assessments to evaluate student learning (14.4)
Authors: Bradley Bryan
Published: June 2021

Student learning within any course of study is a blend of many things, including needs, goals, interests, motivations, skills, knowledge, etc. Both instructors and students bring their own mixture of these qualities with them into the learning and teaching environment. During our time together we will consider the following questions: • How do instructors know they are effectively teaching what their students need to know and do? • How do students know they are effectively learning what they need to know and do to succeed? • Can instructors help students improve their learning? • Can students help instructors improve their teaching? We will discuss techniques regarding aligning assessments to learning outcomes, establishing assessment blueprints, assessment data analysis, and using insights from assessment data in writing publishable papers on the Scholarship of Teaching.
DHA regulates the expression of 15‐PGDH through miR 26 a/b in cholangiocarcinoma (144.12)
Authors: Yao Lu , Han Chang , Wu Tong
Published: June 2021

DHA, the primary effector molecule of omega‐3 polyunsaturated fatty acids, may ameliorate inflammatory diseases and prevent carcinogenesis. However, the precise mechanisms by which DHA influence hepatic carcinogenesis have not been elucidated. Here we report DHA regulates PGE2 deactivator‐15‐PGDH expression through miR26a/b in cholangiocarcinoma. Human cholangiocarcinoma cell lines (CCLP‐1 and TFK‐1) treated with DHA or stably transfected with Fat‐1 gene significantly enhanced 15‐PGDH protein expression while had little effect on its mRNA level. MiR26a/b, which putatively binds to 15‐PGDH 3’UTR, are significantly down‐regulated in DHA‐treated or Fat‐1 expression cells. MiR26a/b overexpression greatly reduced 15‐PGDH protein level. Since miR26a/b locate in intron region of gene family CTDSP, their expression is intensely correlated and regulated by transcription factor C‐myc. We found significant consistency between C‐myc expression and CTDSPL, CTDSP1 gene or miR26a/b levels. Tumor growth assays, in vitro and in vivo, showed Fat‐1 undermined tumor growth while miR26a/b had a reverse effect. This phenomenon could be attributed to that those two factors regulate the 15‐PGDH expression and further affect the PGE2 production. Taken together, our data provide important mechanistic insights and therapeutic implications for utilizing omega‐3 PUFAs for the treatment of human cholangiocarcinoma.

Grant Funding Source: NIH/NCI
Autophagic flux alters hepatic iNOS protein accumulation, functional form, and localization (151.8)
Authors: Loughran Patricia , Sun Qian , Scott Melanie , Stolz Donna , Mastroberardino Pier , Friedman Peter , Watkins Simon , Billiar Timothy
Published: June 2021

The hepatic iNOS response to different stressors varies with the amount, duration, cell type and source of nitric oxide. Autophagy is known to be regulated by inducible nitric oxide synthase (iNOS) and nitric oxide (NO), but little is known about how autophagic flux impacts iNOS protein levels. Regulation of iNOS accumulation is important to understand as many metabolic processes are influenced by oxygenation/nitrosylation mediated by iNOS/NO during inflammation. We have also previously shown that iNOS binding to ezrin radizin moesin binding phosphoprotein 50 (EBP50) allows transport of iNOS to peroxisomes and then degradation via pexophagy. The aim of this study was to identify the role of alterations of autophagic flux on iNOS in hepatocytes after stimulation with cytokine mix (CM: TNF[500U/ml]; IL1[200U/ml]; IFN [100U/ml]). Rat hepatocytes were stimulated with CM +/‐ autophagic inhibitors for 8h. Cells were analyzed by Griess, immunoblot, and confocal microscopy to determine the localization, functional form and accumulationof iNOS. iNOS peroxisomal localization is maximal with the inclusion of 3MA and to a lesser degree the NO inhibitor, L‐NIO. This form of iNOS was determined by immunoblot to be monomeric. Accumulation of iNOS and its reactive products at the various stages of autophagic sequestration is of interest due to the shift of metabolic processes that rely on heme proteins susceptible to modification by oxygen and nitrogen radicals characteristic in surgical models generated in inflammation.

Grant Funding Source: R37‐GM044100 awarded to TRB
Sowing the seeds of integrative health care: implementation of an interprofessional anatomy curriculum for first year students (18.4)
Authors: Titelbaum Anne , Lucas Lynn , Fisher Rebecca
Published: June 2021

The University of Arizona (UA) College of Medicine Phoenix campus opened in 2006 and has since partnered with Arizona State University (ASU) and Northern Arizona University (NAU) to provide an integrated anatomy curriculum for medical, physician assistant, physical therapy, and bioengineering and anthropology graduate students. Divided into two class groups (UA and ASU/NAU), students utilize the same gross anatomy laboratory and dissect the contralateral sides of the same willed body donors. In order to foster collaboration and a greater understanding and appreciation for all members of the health care team, an interprofessional anatomy curriculum was developed for the 2013 fall semester that included a meet the donor session, maintenance of a donor (patient) chart, four clinically oriented workshops, and a ceremony of appreciation. Assessment of the new curriculum included quantitative measures related to workshop activities and exam performance as well as student and instructor feedback. Results indicate that the integrated curriculum increases student engagement and fosters an attitude of interprofessional cooperation and respect, which enhances student support systems and lays the groundwork for future professional development.
Designing an interprofessional learning curriculum for medical and physical therapy students in gross anatomy curriculum (18.5)
Authors: Euerle Terin , Haller Elizabeth , Lachman Nirusha , Pawlina Wojciech , Youdas James , Hellyer Nathan , Krause David
Published: June 2021

Interprofessional collaboration in healthcare is crucial for quality patient care. While it continues to be an expected competency, most educational programs lack interprofessional learning (IPL) experiences. We administered the five‐point Likert Readiness of Interprofessional Learning Scale (RIPLS) to first year medical (MD) and physical therapy (PT) students to evaluate attitudes toward IPL. Students agreed that “For small group learning to work, students need to trust and respect each other” (average of 4.6 out of five points). They also disagreed that learning with other healthcare students is a waste of time or unnecessary (averages of 1.91 and 1.98 respectively).

Clearly students desire IPL experiences, and with this in mind, we established an IPL curriculum for their gross anatomy courses. Students were first grouped into interprofessional teams at a social event. In dissection lab, PT students taught upper limb and MD students taught thorax anatomy. We also conducted IPL anatomy based problem solving sessions in a classroom setting.

Anatomy courses provide an ideal platform for IPL opportunities. Lab and classroom activities enable students to work together through a shared experience of understanding anatomical concepts from different perspectives. By establishing an IPL curriculum, we hoped to increase interprofessional collaboration early in students’ medical education. Our results indicate positive student attitude toward the learning experience.
Mixed methods evaluation of an online undergraduate systemic human anatomy course with laboratory (211.1)
Authors: Attardi Stefanie , Barnett John , Rogers Kem
Published: June 2021

An online section of a Systemic Human Anatomy course was offered for the first time in 2012/13. Lectures for face‐to‐face (F2F) students (n=365) were broadcast in live and archived format to online students (n=40) using virtual classroom software (Blackboard Collaborate). Labs were delivered online by a teaching assistant who manipulated 3D computer models in the virtual classroom. A mixed methods approach is being used to determine the effectiveness of the online format. Student performance measures (4 tests, 24 lab quizzes) were statistically identical between sections. Incoming grade averages were strongly correlated to overall anatomy grade in both F2F (r = 0.70, p < 0.01) and online (r = 0.63, p < 0.01) sections, suggesting that prior academic performance, and not course format, predicts performance in anatomy. Interviews (22 online; 38 F2F students) and surveys (270 F2F students) regarding perceptions of the learning experience were conducted following a cross‐over period that exposed students to both formats. Survey results indicate that while students preferred online lectures (52%), F2F labs were preferred (85%). Online lectures gave students the benefit of reviewing archived sessions, while F2F labs allowed for better student‐teacher communication. A content analysis of interview transcripts is being undertaken to generate grounded theory about the strengths and weaknesses of the online course.

Grant Funding Source: Departmental funding
Necessity is the mother of educational innovation: a journey of discovering and developing electronic pedagogical tools for gross anatomy and histology (215.2)
Authors: Wisco Jonathan
Published: June 2021

From pencil and paper, to chalk and chalkboard, to transparencies and overhead projectors, to Microsoft PowerPoint and computers with HDTV, to the Internet and beyond, technology has played a major, symbiotic role in the innovation of pedagogy. Many technologies have advanced pedagogy, but it can be argued that pedagogies have also advanced technologies. Whether technologically advanced or simplistic in design, teaching innovations have the same goal: to advance the effectiveness and efficiency of the learning process. Some innovations are tailored to a specific, in‐home audience, but others may be appropriate for translational education, which we define as the commercialization of educational tools for a public audience. We discuss our experience with developing technological and non‐technological innovative methods over the past 20 years that have helped to transform traditional gross anatomy and histology pedagogy to an interactive flipped‐classroom style; and we share our decision making process in determining which innovations should be published and which should be registered for possible commercial distribution.
Preparing minority scientists and engineers (220.1)
Authors: Summers Michael , Hrabowski Freeman
Published: June 2021

African Americans comprise about 14 percent of college students in the U.S. but earn only ~ 7 percent of bachelors degrees and ~3 percent of doctoral degrees in STEM fields. We have developed a system for diversifying our undergraduate and graduate STEM programs that focuses on high expectations and achievement. More than 1200 students have participated in the undergraduate Meyerhoff Scholars Program since its inception in 1988. More than 90% of the participants who graduated received STEM degrees, with > 40% matriculating to top PhD or MD‐PhD programs, 20% to STEM Masters programs, and 20% entered professional (mainly MD) programs. The talent pool is large: we typically receive more than 2,200 nominations and more than 500 applications annually (about half URM students), mainly from Maryland and the Mid‐Atlantic area. URM participation in STEM PhD programs has also increased dramatically over the past 15 years (from 3% to 17%), and URM PhD production has increased from 3 graduates in the 10 years preceding our program to more than 50 graduates over the past 10 years. The program currently includes 80 URM doctoral students, and the retention rate over the past 8 years is greater than 90%. More than 40 former UMBC undergraduate and graduate students now hold academic faculty positions in the U.S. The program components of the undergraduate and graduate programs should be replicable at other interested academic institutions.
Prefolding and quality control of OMPs in periplasmic space (224.1)
Authors: Zhao Xin Sheng
Published: June 2021

The outer membrane proteins (OMPs) of Gram‐negative bacteria have to be translocated through the periplasmic space before reaching their final destination. The aqueous environment of the periplasmic space and high permeability of the outer membrane engender such a translocation process inevitably challenging. In Escherichia coli, although SurA, Skp and DegP have been identified to function in translocating OMPs across the periplasm, their precise roles and their relationship remain to be elucidated. In addition, DegP is said to have protease/chaperone dual function. However, whether DegP truly performs chaperone activity has long been under debate, let alone how this dual function is regulated. Our single molecule detection study has revealed the mechanism on the collaboration among SurA, Skp and DegP. We have also found that DegP indeed processes chaperone‐like activity. The dual function is regulated by both the substrate conformation and oligomerization status of DegP itself. Based on our results, a model on how OMPs are safeguarded and quality controlled in the periplastic space is proposed.
Discovery of novel enzyme activities and metabolic pathways (227.3)
Authors: Gerlt John
Published: June 2021

The Enzyme Function Initiative (EFI) is a Large‐Scale Collaborative Project (U54GM093342) that is devising strategies and tools to facilitate the discovery of the in vitro activities and in vivo metabolic functions of enzymes discovered in genome projects. This talk will provide examples of how 1) bioinformatic and computational tools can be used to predict novel enzymatic functions and pathways; and 2) enzymology, microbiology, and metabolomics can be used to verify those functions and place them in metabolic context. Supported by NIH U54GM093342.
I write so you don't have to (233.2)
Authors: Gordon Debra
Published: June 2021

In this presentation, a veteran medical writer will discuss the life of a medical writer, including pathways into the profession, types of work available, salary ranges, joys and frustrations. The focus will be on medical writing as an alternative career path for scientists. Participants will learn about the skills required for both in‐house and freelance positions and resources for education and training.
Signals and blockers in karyopherin‐mediated transport (238.1)
Authors: Chook YuhMin
Published: June 2021

Karyopherinβ (Kap; Importin or Exportin) proteins mediate nucleocytoplasmic transport. Individual Kaps transport unique sets of cargos through recognition of distinct nuclear localization or export signals (NLSs or NESs). Although there are 19 known Kaps in humans, only four classes of signals are known at this time: the short basic classical‐NLS, the PY‐NLS, a distinct basic NLS with the X motif and the leucine‐rich NES, which are recognized by Kapα/Kapβ1, Kapβ2 (or Transportin), Kap121 and CRM1, respectively. Most other Kaps recognize diverse sequences that occlude the identity of their signals. Some nuclear import cargos are also transported into the nucleus by several Kaps and it is unclear if the cargos contain a single NLS that is recognized by multiple Kaps or contain multiple NLSs. Studies to identify new classes of NLSs and to understand simultaneous recognition of cargos by multiple Kaps are ongoing. The karyopherin CRM1 is the general nuclear export factor that exports hundreds of NES‐containing proteins from the nucleus. Structures of CRM1 bound to lactone polyketide inhibitors like Leptomycin B and to small drug‐like inhibitors KPT‐SINEs have recently been solved. Mechanisms of CRM1 inhibition by both classes of compounds provide insights into differences in toxicity.
Avocado consumption decreases LDL particle number and small, dense LDL cholesterol in overweight and obese subjects (249.8)
Authors: Wang Li , Fleming Jennifer , Kris‐Etherton Penny
Published: June 2021

We evaluated the effects of a moderate fat diet high in MUFA provided by one avocado per day on lipoprotein particle number and lipoprotein subfractions compared to a moderate fat diet (with equivalent fatty acid content) and a lower fat diet. A randomized 3‐period cross‐over, controlled feeding study was conducted in healthy, overweight and obese men and women (n=45, 21‐70 years, BMI 25‐35 kg/m2) with LDL‐C in the 25‐90th percentile. Following a 2 wk run‐in average American diet (AAD), 3 isocaloric diets (5 wk each) were fed: 1) a lower‐fat diet (LF: 24% fat, 59% CHO, 16% PRO); 2) a moderate fat diet providing one HASS avocado/day (136g) (AV: 34% fat, 49% CHO, 16% PRO); and 3) a moderate fat diet using high oleic acid oil as the main source of MUFA (MF: 34% fat, 49% CHO, 16% PRO). Fasting blood samples were collected after each diet. Lipoprotein subfractions were measured by the vertical auto profile (VAP), lipoprotein particle number was measured directly by proton nuclear magnetic resonance spectroscopy (NMR LipoProfile). Compared to the AAD, only the AV diet significantly decreased LDL particle number (LDL‐P) (‐5.8%, p=0.006). The MF diet did not change LDL‐P significantly (‐2.9%, p=0.2), and there was a trend for an increased LDL‐P on the LF diet (3.8%, p=0.08). We observed similar reductions of cholesterol in large, buoyant LDL particles (LDL1+2). However, only the AV diet significantly reduced cholesterol in small, dense LDL (sdLDL) (LDL3: ‐8.7%, P<0.01; LDL3+4: ‐5.6%, P=0.05) and the sdLDL level after consumption of AV diet was significantly lower than MF diet (p<0.05). The change in LDL‐P by diets was significantly correlated with the change in sdLDL (r=0.58, p<0.001) and apoB (r=0.69, p<0.001) but was not correlated with large, buoyant LDL (r=0.06, p=0.5). In conclusion, incorporation of one avocado/day as part of a healthy, moderate fat diet has beneficial effects beyond its MUFA content on decreasing both LDL‐P and sdLDL in overweight and obese subjects. ClincalTrials.gov Identifier: NCT01235832.