bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histology of liver oxidative stress in an obese mouse model of nonalcoholic fatty liver disease treated with Ω3 fatty acids, TNFα‐receptor blocker, or curcumin (1116.1)
Authors: Likhitsup Alisa , Alba Laura , Herndon Betty , Mansour Ahmad , Zia Hamid , Molteni Agostino 
Published: June 2021

Nonalcoholic fatty liver disease (NAFLD) origins are unclear but progression involves inflammation and correlation with obesity and insulin resistance. Advanced glycation end products (AGEs) are oxidative stress markers that induce inflammation and release reactive oxygen species. AGEs play key roles in the development of vascular inflammation of many types. Tissue accumulation of AGEs is identified by receptor staining (RAGE) . This study evaluated dietary and drug intervention to interrupt RAGE staining in a model of NAFLD. OB/OB leptin knock‐out mice (10/group) received clinical levels of curcumin, Ω3 fatty acids, TNF‐α receptor blocker or control chow for 8 wk. Liver steatosis was scored at 400x. Liver was also stained with PAS for glycoproteins and RAGE antibody. The Image J program was used for quantification of RAGE stain percentage. By H&E, significant reduction of liver fat was seen in groups receiving curcumin or Ω‐3 fatty acids compared to controls. By RAGE stain, interventions reduced AGES from 38.1% (untreated) to 26.4% (TNF blocker) and 26.8% (Ω3). p value, 0.15. This pilot experiment suggests that AGEs contribute to the development of nonalcoholic fatty liver disease and/or the progression of NAFLD to nonalcoholic steatohepatitis. The progression, in our model, is interrupted by food additives or an anti‐inflammatory. Studies to expand this approach to interrupt NAFLD are planned.Grant Funding Source: Funding: St Luke’s Hospital Foundation grant to LA
Evaluation of novel inflammation biomarkers GlycA and GlycB in overweight and obese adolescents with NAFLD (1116.9)
Authors: Jin Ran , Otvos James , Le Ngoc‐Anh , Vos Miriam 
Published: June 2021

Objective: Cardiovascular disease (CVD) is a leading cause of increased mortality in patients with nonalcoholic fatty liver disease (NAFLD). One of the proposed roles of NAFLD in the pathogenesis of CVD is through the systemic release of inflammatory mediators. GlycA and GlycB are two novel inflammation biomarkers that are both associated with future CVD and diabetes. However, these markers have not been measured in pediatric NAFLD and their relationships to inflammation in NAFLD are unknown.Methods: We examined 37 Hispanic overweight and obese adolescents who had presumed NAFLD (hepatic fat > 5% by MRI). GlycA and GlycB were measured by nuclear magnetic resonance (LipoScience, Raleigh, NC).Results: In our cohort, both GlycA and GlycB were significantly correlated with hs‐CRP (r=0.744, p<0.001 and r=0.694, p<0.001). GlycA, but not GlycB, was also associated with BMI z‐score (r=0.334, p=0.043) and leptin (r=0.427, p=0.026). However, no significant correlations of GlycA and GlycB were observed with hepatic fat content and liver enzyme ALT.Conclusion: Our preliminary data suggest that in pediatric NAFLD, GlycA and GlycB are associated with systemic inflammation and may have applications similar or complementary to hs‐CRP and other inflammatory biomarkers.Grant Funding Source: Supported by NIH K23 DK080953 (Vos)
Gastric antral organoids are sustained by intrinsic Notch signaling (1119.1)
Authors: Gifford Gail , Demitrack Elise , Spence Jason , Simeone Diane , Samuelson Linda 
Published: June 2021

The gastric epithelium undergoes constant turnover, with adult stem cells replenishing mature cell types to maintain homeostasis. The Notch signaling pathway is known to regulate gastric epithelial proliferation and differentiation, but little is known about mechanism. In this study we tested the hypothesis that Notch signaling is intrinsic to the epithelium and supports antral stem cell function. Mouse and human gastric organoids were used as a model to define epithelial Notch signaling. Organoids were established from antral glands and maintained in 3D culture in media containing EGF, Noggin, R‐spondin1 and Wnt3A. Gastric organoids were highly proliferative and contained differentiated gastric cell types, suggesting that they are sustained by antral stem cells. Notch pathway expression was determined by qRT‐PCR and function was tested using the Notch inhibitor DAPT. Pathway inhibition reduced cellular proliferation and organoid growth, and lowered the efficiency of organoid re‐establishment. Together these studies show that Notch signaling is intrinsically established by gastric epithelial cells in vitro to promote organoid proliferation and to maintain stem cell function.Grant Funding Source: Supported by NIH grants T32GM008322‐21/22 (GBG), T32DK094775‐02 (GBG), P01‐DK062041 (LCS)
Inhibition of a hyperpolarization‐activated inward current (Ih) facilitates cold‐activation of locus coeruleus neurons of bullfrogs, Lithobates catesbeianus (1128.2)
Authors: Santin Joseph , Watters Kayla , Hartzler Lynn 
Published: June 2021

Mechanisms that determine neuronal excitability under variable temperatures are unclear. In bullfrogs, cooling increases action potential (AP) firing in locus coeruleus (LC) neurons. We used whole‐cell patch clamp electrophysiology in brainstem slices to test whether inhibition of a hyperpolarization‐activated current (Ih) promotes increases in LC neuron excitability during cooling from 20°C to 10°C. Cooling reduced maximum Ih amplitude from ‐211.5±55.0pA to ‐69.5±7.8pA (P<0.05) and shifted the voltage at half maximal activation (V50) from ‐69.9±11.4mV to ‐92.9±5.8mV (P<0.05). Application of 2mM CsCl inhibited Ih (‐142.8±53.3pA vs. ‐3.3±14.3pA; P<0.05), and may potentiate AP firing increases induced by cooling (increase in AP firing; control: 0.9±0.5Hz vs. Cs+: 2.2±1.6Hz; P=0.2; n=3). Elevating intracellular cyclic AMP to 100μM increased the proportion of activated Ih near resting membrane potential (Vm ) at 20°C and 10°C (V50 =‐63.7±9.5mV and ‐71.3±10.2mV); yet, activation of Ih suppressed the firing increase elicited by cooling (0.2±0.7Hz decrease vs. increased firing in controls; P=0.06; n=5). Our data suggest inhibition of Ih enhances LC excitability during cooling and implicate Ih as a temperature‐sensitive mechanism that modulates neuronal excitability during changes in brain temperature. Given extensive involvement of LC neurons in physiological functions, understanding the regulation of LC neuron excitability during temperature changes has broad ramifications.Grant Funding Source: Supported by NSF IOS 1257338
Integrative bioinformatical analysis of clear cell renal cell carcinoma (1138.10)
Authors: Butz Henriett , Nofech Mozes Roy , Rotondo Fabio , Kovacs Kalman , Patocs Attila , Szabo Peter , Yousef George 
Published: June 2021

Background. Clear cell renal cell carcinoma (ccRCC) is the most common adult neoplasm of the kidney. Better understanding of the molecular pathogenesis may lead to new therapeutic options.Aim. Our aim was to assemble transcriptomic, proteomic and miRNA data using an integrative approach.Methods. Available mRNA, miRNA and protein data were collected from 593 ccRCC and 389 normal kidneys. We performed pathway analysis. TCGA database and immunohistochenmitry were used for validation. Functional analyses were undertaken on cell line model.Results. Metabolic processes were the most significant signalling pathways, and we validated Aryl‐Hydrocarbon Receptor (AHR) signalling to be involved in tumorigenesis. Using network analysis, we identified GRHL2 as diagnostic marker and drug target. KIAA0101 was found to enhance tumor cell migration and invasion. We also demonstrated that KIAA0101 overexpression correlates with poor prognosis and may represent a diagnostic marker. We identified miR‐139‐5p as the miRNA mostly affecting the network targeting ZEB1, TCF4, ETS1 and CCND2.Conclusions. Using integrative bioinformatical analyses we identified the most characteristic pathways and molecules. Some of them may be applicable as biomarkers or new, potential drug targets.
Angiotensin II stimulation alters vasomotor response to adenosine agonist in mesenteric artery: ole for A1 and A2B receptors (1140.4)
Authors: Yadav Vishal , Nayeem Mohammed , Tilley Stephen , Mustafa S. Jamal 
Published: June 2021

Angiotensin II (Ang II) is one of the major mediators that lead to the development of hypertension. However the signaling pathway that leads to hypertension are not well understood. Ang II activates several intermediate molecules which may alter localized signaling events to further regulate vascular tone that may contribute to the development of hypertension. Adenosine is the important metabolite which controls local blood flow through activation of four subtypes of adenosine receptors (ARs). Previously, we have reported that A1AR is the major determinant of contractions while A2B produces relaxation in mouse mesenteric arteries (MAs). In this study, we sought to investigate whether acute exposure to Ang II alters vasomotor responses to non‐selective AR agonist, NECA, in MAs. We used DMT myograph system for muscle tension measurement in isolated resistant MAs. We have found that acute exposure (15 min) to Ang II (10 nM) altered dose‐dependent vasodilatation to NECA producing significantly higher contractions (~37%) in MAs. Notably, these higher contractions were absent in MAs from A1 KO mice. Interestingly, we have also found that NECA induced relaxation in MAs from A1 KO mice were inhibited (~10%) after exposure to Ang II. Importantly, in MAs from WT mice, we have found that pre‐treatment with HET0016 (10 µM, 20HETE inhibitor) significantly inhibited Ang II and NECA‐induced contractions. In addition, in MAs from A2B KO mice, NECA produced contractions which were significantly higher after Ang II stimulation. These contractions were considerably inhibited by HET0016 and completely blocked by DPCPX (1 µM, A1 antagonist). Moreover, Ang II contractions were higher in MAs from A2B KO mice (p<0.05). Taken together, our data indicate that acute Ang II stimulation enhances A1 and reduces A2B‐dependent signaling in MAs. Thus, these altered A1 and A2B dependent signaling pathway may contribute to the development of hypertension.Grant Funding Source: Supported by HL 094447, HL027339, HL114559 and AI 096139
Effect of alcohol on clopidogrel metabolism: Identification of a new alcohol‐dependent metabolic pathway (1141.14)
Authors: Hu Zheyi , Laizure S. , Herring Vanessa , Parker Robert 
Published: June 2021

The majority of orally administrated clopidogrel, a platelet aggregation inhibitor, is hydrolyzed by human carboxyesterase‐1 (CES1) to the inactive clopidogrel acid metabolite with the remainder metabolized by CYP450 enzymes into the inactive intermediate, 2‐oxo‐clopidogrel, that is further oxidized to the active thiol metabolite. The objective of this study was to determine the effect of alcohol, a known CES1 inhibitor, on the metabolism of clopidogrel. Clopidogrel metabolism was studied in human recombinant CES1, human liver S9 fractions (HLS9), and in plasma‐carboxylesterase knockout mice. Liquid chromatography/triple quadrupole mass spectrometry (LC‐MS/MS) was utilized for the quantification of clopidogrel and its metabolites. Alcohol‐derived metabolites were identified by product ion scan using LC‐MS/MS. The addition of alcohol resulted in the formation of the transesterified clopidogrel metabolite, ethyl‐clopidogrel in CES1 and HLS9 incubations. Similar to clopidogrel, ethyl‐clopidogrel was metabolized to ethyl‐2‐oxo‐clopidogrel followed by the subsequent oxidation to thiol metabolites in HLS9. Co‐administration of alcohol (3 g/kg) and clopidogrel (25 mg/kg) to plasma‐carboxylesterase knockout mice resulted in a reduced systemic exposure to clopidogrel acid (control AUC0‐8h 39 µg·h/mL versus alcohol AUC0‐8h 20 µg·h/mL) and the formation of transesterified metabolites. Alcohol significantly inhibits the hydrolysis of clopidogrel to clopidogrel acid, and produces transesterified metabolites of clopidogrel, 2‐oxo‐clopidogrel, and the active metabolite.Grant Funding Source: Supported by National Institutes of Health National Institute of General Medical Sciences
Overfeeding of saturated fat and fructose induces alterations of hepatic fatty acid composition in rats (1142.10)
Authors: Tricoli de Souza Verônica , Priviero Fernanda , Claudino Mario , Rocha Thalita , Messias Marcia , Carvalho Patricia 
Published: June 2021

Fatty acids (FAs) are essential components of lipids and exhibits important biological functions. The analysis of FAs profiles in tissues has become increasingly important as end points in understanding the roles of FAs in physiological or pathological processes. This study aimed to describe the changes in fatty acid composition of the liver lipids caused by excessive consumption of high saturated fat diet associated with fructose in Wistar rats. Rats were fed ad libitum with chow (3 Kcal/g; 2.3% soybean oil) or high‐fat diet (5.5 Kcal/g; 30% lard and 4% soybean oil) associated with fructose (100mg/mL) for 12 weeks. The lipids of liver homogenates were extracted with chloroform‐MeOH and the fatty acids methyl esters were analyzed by gas‐chromatography. Fatty acid composition of the liver was found to shift from a pattern dominated by saturated (SFA, C16:0), monounsaturated (MUFA, 18:1 n‐9) and polyunsaturated FA (PUFA, C18:2 n‐6). Oleic acid (C18:1 n‐9) increased (approximately 36%, p=0.0001) whereas PUFA (C20:4 n‐6 and C22:6 n‐3) and SFA (18:0) decreased in the liver of animals fed with high saturated fat and fructose. This difference in the fatty acid profile in the liver is reflected in the increased MUFA/SFA ratio. The data indicated that the overfeeding of high saturated fat and fructose diets alter the hepatic fatty acid composition and the oleic acid is greatly accumulated in the liver.Grant Funding Source: FAPESP ‐ 2012/11050‐6
Establishment and evaluation of physical and psychological stress‐induced PTSD animal model (1144.7)
Authors: Yu Bin , Zhang Xueqiong , Cui Xiangyu , Cui Suying , Zhang Yonghe 
Published: June 2021

It is well known that not only direct recipients but also the witnesses of traumatic events could suffer PTSD. However, there are no recognized animal models to mimic psychological stress induced PTSD. So we tried to establish psychological stress induced PTSD by communication box. Physical stress (foot‐shock, FS) was generated by receiving foot‐shock directly which mimicked the direct recipients of trauma, while psychological stress (non‐foot shock, PS) was generated by exposure to emotional responses without direct physical stress, for example, the visual and auditory stimuli that arose from foot shock stress rats, to mimic the witnesses of trauma. Serum corticosterone levels were increased three times 30 minutes after stress in both FS and PS rats, this result suggested that the modeling process was intense threat for both FS and PS rats. 21 days after trauma, we evaluated whether rats presented PTSD symptoms, like anxiety, high vigilance, avoiding trauma‐related cues and strong memory of trauma, by using elevated plus maze/ open field test, acoustic startle response test, ball (trauma related cue) burying test, and freezing time (re‐exposed to communication box) test. The results showed that about 40% FS rats and 30% PS rats could simulate PTSD clinical symptoms, and this animal model could be helpful to explore PTSD mechanism specifically and comprehensively.Grant Funding Source: This study was supported by the NNSFC (No.81173031,81202511 and 81302746).
The effect of S‐equol in attenuation of cell injury under oxidative stress in human umbilical vein endothelial cells (1146.1)
Authors: Zhang Ting , Mi Mantian , Liang Xinyu , Shi Linying 
Published: June 2021

Objective To study the protective effects of S‐equol on oxidative stress‐induced cell injury in human umbilical vein endothelial cells (HUVECs).Method HUVECs were used in this study. To establish the oxidative stress‐induced cell model, effect of H2O2 on cell viability was detected by CCK‐8 assay. The same method was used to detect the effect of S‐equol on cell apoptosis using flow cytometry and TUNEL assay. Western blot assay was used to detect the effect of S‐equol on caspase‐3 and Bcl‐2 protein expression. SOD, CAT and GSH levels were determined using SOD, CAT and GSH kits.Results The IC50 of H2O2 was 650μM. S‐equol (1 nM, 10 nM, 100 nM) significantly dose‐dependently inhibited H2O2‐induced decrease of cell viability in HUVECs. S‐equol (100 nM) significantly inhibited cell apoptosis in e HUVECs under oxidative stress, and reduce the number of TUNEL labeling‐positive cells. S‐equol (100 nM) up‐regulated Bcl‐2 protein expression level and significantly inhibited caspase‐3 protein expression increased by H2O2. S‐equol dose‐dependently up‐regulated SOD, CAT and GSH levels, and inhibited H2O2‐induced decrease in SOD, CAT and GSH levels.Conclusion Low concentration (physiological concentration) of S‐equol attenuated oxidative stress‐induced cell injury in HUVECs, which relates to the increase in SOD, CAT and GSH level.Grant Funding Source: Supported by the National Natural Science Foundation of China (No. 81102129)
Study on mechanisms of action of liuwei dihuang decoction with proteomic and metabonomic techniques (1147.3)
Authors: Jiang Ning , Zhou Wenxia , Zhang Yongxiang , Yan Xianzhong , Zhang Qi 
Published: June 2021

Objective: Liuwei Dihuang (LW) decoction are classical traditional Chinese medicinal prescription for anti‐aging. Senescence‐accelerated mouse (SAM) is a good model to study mechanisms of aging. To better understand the mechanisms of actions of LW on antiaging, systematic techniques, including proteomic and metabonomic methods, were employed in this study. Method: SAM‐prone/8 (SAMP8) were orally administered with LW for consecutive 30 days. Two dimensional electrophoresis (2‐DE) based proteomic technique and 1H nuclear‐magnetic‐resonance‐spectroscopy (NMR) based metabonomic technique were employed to investigate the proteins and metabolites that LW acted on. Results: The expressions of serum proteins in SAMP8 before and after LW treatment were quite different. There were 21 differentially expressed proteins after LW treatment, which were related to the functions of transcriptional control, cytoskeleton, cell signaling, etc.. Principle components analysis of 1HNMR spectra showed clear separation between serum samples of SAMP8 before and after LW treatment. Metabolites that LW affected were glucose, lactate, high density lipoprotein, etc. related to glucose, lipids, lipoproteins and amino acids metabolism. Conclusion: These findings suggested abnormal serum protein expressions and metabolism disequilibrium in SAMP8 caused by aging and the mechanisms of actions of the LW may lie in restoring of the normal condition of specific proteins and metabolism equilibrium in SAMP8.Grant Funding Source: Supported by grants from the National Natural Science Foundation of China 30701073, 90709012
Interleukin‐6 modulates transforming growth factor‐β receptor II function and receptor compartmentalization in epidermal keratinocytes (1148.11)
Authors: Chastain Lerin , Cottrell Mackenzie , Ihnat Mike , Gallucci Randle 
Published: June 2021

IL‐6 and TGF‐ β mediate a broad range of biological responses including growth, differentiation, and migration. We have previously shown that IL‐6 modulates Tgf‐β1 expression in fibroblasts, however; what role IL‐6 plays concerning TGFβ‐RII expression and function in skin cells is unknown. Therefore, the aim of this study was to investigate the mechanism by which IL‐6 might modulate TGFβ‐RII in skin.Skin from WT and IL‐6 over‐expressing mice and IL‐6 treated keratinocyte cultures were analyzed for TGF‐βRII expression via immunohistology and western blot. TGF‐βRII function and activity was assessed by cell migration, proliferation, and Smad2/3 phosphorylation. TGF‐βRII membrane localization was determined by co‐immunoprecipitation. IL‐6 overexpression and IL‐6 treatment increased TGF‐βRII expression in epidermis and isolated keratinocytes respectively. IL‐6 treatment of keratinocytes augmented TGF‐β1 function as demonstrated through increased migration and decreased proliferation. TGF‐ βRII is down regulated when located within caveolin rich lipid rafts and co‐IP showed with IL‐6 treatment, association between TGF‐ βRII and caveolin increased resulting in decreased Smad2/3 phosphorylation. These results suggest that IL‐6 regulates TGF‐ β1 function by modulating TGF‐ βRII expression and increasing trafficking of the receptor to lipid raft pools.Grant Funding Source: NIH/NIGMS R01GM67745
Atorvastatin inhibits induction of chemokine receptor CXCR7 to reduce macrophage migration (1148.12)
Authors: Liu Yiwei , Ma Wanshu , Crocker Laura , Shen Jianzhong 
Published: June 2021

We recently reported that the new SDF‐1 receptor CXCR7 is induced during monocyte‐to‐macrophage differentiation in the context of atherosclerosis. This study aimed to determine whether CXCR7 induction during macrophage differentiation is inhibited by statins. Here we show that atorvastatin inhibited CXCR7 mRNA and protein expression in THP‐1 macrophages, without affecting CXCR4 expression. Suppression of CXCR7 expression was reversed by supplementation with mevalonate. Inhibition of squalene synthase, the enzyme committed to cholesterol biosynthesis, partially decreased CXCR7 induction. However, the geranylgeranyl transferase inhibitor, GGTI‐286, the farnesyl transferase inhibitor, FTI‐276, and the Rho kinase inhibitor, Y‐27632, all failed to mimic the effect of atorvastatin, suggesting that the protein prenylation pathways are not critical. Interestingly, the effect of atorvastatin was not fully mimicked by other statins. Furthermore, activation of CXCR7 by SDF‐1, TC14012, or I‐TAC all prompted macrophage migration, which was suppressed by atorvastatin treatment, but not by the CXCR4 antagonist. We conclude that atorvastatin modulates macrophage migration by down‐regulating CXCR7 expression, suggesting a new CXCR7‐dependent mechanism of atorvastatin to benefit atherosclerosis treatment independent of its lipid lowering effect.Grant Funding Source: American Heart Association ─ National SDG grant 12SDG8850011
Selective estrogen receptor modulators resveratrol, genistein, tamoxifen, pterostilbene and clomiphene downregulates the expression of minute‐double minute2 (MDM2) and vascular endothelial growth factor (VEGF) genes in SNB‐19 glioblastoma cells (1148.9)
Authors: Bhatt Vrushank , Ratna Warren 
Published: June 2021

Glioblastoma multiforme is the most common primary neoplasm of the brain. Our studies using SNB‐19 glioma cells have shown that selective estrogen receptor modulators (SERMs) at pharmacological concentrations modulate the expression of minute double minute (mdm2) and vascular endothelial growth factor (VEGF) genes. The SNB‐19 glioblastoma cells were treated with increasing concentrations of SERMs such as resveratrol, genistein, tamoxifen, pterostilbene and clomiphene, and their effects were studied using RNA and protein analyses. The cell viability studies on cells exposed to the above SERMs showed a concentration dependent cell growth arrest and cell death by apoptosis. Reverse‐Transcription Polymerase Chain Reaction (RT‐PCR) studies indicated decreased levels of transcription of mdm2 and VEGF mRNA. Western blot analysis showed a corresponding decrease in the expression of the VEGF protein. The above results show the down‐regulation of the mdm2 and VEGF expression and the induction of cell death in SNB‐19 glioma cells due to exposure to the above SERMs, indicating their potential use in causing the regression of glioblastomas.Grant Funding Source: Supported by science research funds from Long Island University, Brooklyn, NY