bluuebunny/crossref_metadata_2025_split huggingface dataset:
Correlation of beta‐arrestin1 levels in mononuclear leukocytes with depression scores for women with premenstrual dysphoric disorder (1144.2)
Authors: Nayyar Tultul , Alam Farzana , Richie William , Ansah Twum , Bailey Rahn 
Published: June 2021

OBJECTIVE: Depression is very common in reproductive women with premenstrual dysphoric disorder (PMDD), a severe form of premenstrual syndrome (PMS). This study examined whether a measurement for beta‐arrestin1 levels in peripheral blood mononuclear leukocytes (PBMC) could aid in the diagnosis of PMDD and support the results from questionnaires. DESIGN AND METHODS: 18‐42 years old non‐pregnant women at the luteal phase with the symptoms of PMS/PMDD, but not taking any antidepressants, were studied. The protein levels of beta‐arrestin1 in the PBMC were determined by ELISA and compared with the Hamilton Depression Rating Scale scores. The different criteria for Axis 1 mental disorders were compared. RESULTS: The criteria for Axis 1 mental disorders were significantly higher and beta arrestin1 protein levels in mononuclear leukocytes were significantly lower in women with depression (PMDD) as compared to no‐depression (PMS) group. The reduction in beta arrestin1 protein levels was significantly correlated with the severity of depressive symptoms. CONCLUSION: These findings support the implication of beta‐arrestin1 protein in the pathophysiology of depression in women with PMDD. Beta‐arrestin1 measurements in women with PMDD may potentially serve for biochemical diagnostic purposes and might be useful as evidence‐based support for questionnaires.Grant Funding Source: U54 MD 007593 (NIMHD)
Developmental differences in the effect of chronic corticosterone treatment in pre‐ and post‐pubescent female juvenile rats (1144.6)
Authors: Nickle Tyler , Middlemas David 
Published: June 2021

The aim of this study was to determine the effect of chronic corticosterone (CORT) administration on pre‐pubescent and post‐pubescent female rats in order to develop a rat model of depression in these age groups. Rats were administered daily injections of CORT (40 mg kg‐1) for twenty‐one days. Depressive‐like behavior was assessed using the modified forced swim test (FST) and the sucrose preference test (SPT). Immunohistochemistry was used to determine changes in mitogenesis and survival of newborn neurons in the dentate gyrus of the hippocampus. Chronic CORT treatment produced significant changes in the FST for post‐pubescent rats, but not pre‐pubescent rats. Conversely, CORT produced significant changes in the SPT for pre‐pubescent rats, but not post‐pubescent rats. Ongoing investigation will determine the effect of CORT on mitogenesis and survival of newborn neurons in the dentate gyrus of pre‐pubescent and post‐pubescent female rats. These results suggest that CORT may induce depressive‐like behavior in female pre‐pubescent and post‐pubescent rats; however, the physiological effect is manifested differently at different stages of rat development.Grant Funding Source: Supported by AT Still University of Health Sciences grant and the Warner‐Fermaturo Fund
PDE11A negatively regulates lithium responsivity in mice possibly due to an interaction with AKT/PKB (1144.8)
Authors: Kelly Michy , Pathak Geetanjali , Hartman Alexander 
Published: June 2021

Lithium is a first line mood stabilizer for patients with bipolar disorder, but for unknown reasons approximately half of all patients do not respond to lithium. Lithium responsiveness in patients with bipolar disorder has been genetically associated with single nucleotide polymorphisms (SNPs) in Phosphodiesterase 11A (PDE11A), a unique enzyme that breaks down cAMP and cGMP only in the hippocampus. These PDE11A SNPs are intronic; therefore, the objective of this study was to determine if decreasing PDE11A expression was sufficient to increase lithium responsivity in mice. We measured PDE11A expression by in situ hybridization and Western blot in the hippocampus of inbred mouse strains known to respond well (C57BL/6J), moderately (129S6) or poorly (Balb/cJ) to lithium. PDE11A mRNA and protein levels were lowest in C57BL/6J mice, the strain most responsive to lithium, and highest in Balb/cJ mice, the least responsive strain. Consistent with these observations, PDE11A KO mice given 0.4% lithium chow for 3 weeks exhibited greater lithium responsivity relative to wild‐type littermates in tail suspension, an anti‐depressant predictive assay, and in amphetamine hyperlocomotion, an anti‐manic predictive assay. Lithium does not directly affect PDE11A catalytic activity; however, in vitro experiments suggest that PDE11A may influence lithium responsiveness due to an interaction with AKT, an enzyme that is required for lithium’s behavioral effects. AKT is predicted to phosphorylate PDE11A at several residues, including serines 117 and 162. Phosphomimic mutations of serines 117 and 162 shift the subcellular distribution of PDE11A away from a punctate, membrane enriched compartmentalization towards a more diffuse, cytoplasmic localization. Together, these findings suggest that PDE11A negatively regulates lithium responsiveness by inhibiting the activation of AKT but that AKT can feed back and release this brake by phosphorylating PDE11A, thereby removing PDE11A from the membrane.Grant Funding Source: Research supported by a PhRMA Foundation and the University of South Carolina
Dihydromyricetin attenuates high glucose‐induced oxidative stress in human umbilical vascular endothelial cells (1146.2)
Authors: Zhang Ting , Mi Mantian , Liang Xinyu , Shi Linying 
Published: June 2021

Objective To investigate the effects of Dihydromyricetin on high glucose‐induced cell damage in human umbilical vascular endothelial cells (HUVECs).Methods HUVECs were cultured in 5.56 mmol/L glucose as normal control, 5.56mmol/L glucose plus 27.8mmol/L mannitol as an osmotic control, or 33.36 mmol/L high glucose for 48 hours in the absence and presence of Dihydromyricetin, cell viability and apoptosis were detected by CCK‐8 assays and annexin V‐FITC/PI double staining using flow cytometry, respectively. In addition, the intracellular reactive oxygen species (ROS), malondialdehyde (MDA) content and the activity of superoxide dismutase (SOD) and catalase (CAT) in the culture supernatants were measured.Results High concentration of glucose significantly decreased cell viability and the activity of SOD and CAT, while increased the apoptotic rate, the intracellular ROS and the MDA content in HUVECs (P<0.05). Dihydromyricetin pretreatment significantly inhibited the decrease in cell viability and the increase in cell apoptosis induced by high glucose. The ROS and MDA levels were reduced while the SOD and CAT levels were increased after pretreatment with Dihydromyricetin following high glucose stimulation.Conclusion Dihydromyricetin attenuated high glucose‐induced oxidative stress in HUVECs.Grant Funding Source: Supported by The National Natural Science Foundation of China (81102129)
RNA‐guided transcriptional activation of TIMP2 utilizing CRISPR:Cas9 (1148.13)
Authors: Duellman Tyler , Warren Chris , Yang Jay 
Published: June 2021

The clustered, regularly interspaced palindromic repeat (CRISPR) system is capable of genome‐specific targeting utilizing the CRISPR associated 9 (Cas9) endoribonuclease and effector targeting by a single guide RNA (sgRNA). We show that fusion of the transcriptional activating viral protein, VP64, to an enzymatically dead form of Cas9 (dCas9) is capable of enhancing gene‐targeted transcriptional activity. Using a luciferase reporter assay we show that a N‐terminal fusion of VP64 to dCas9 (dCas9.nVP64) results in higher transcriptional activity compared to the C‐terminal or flanked VP64 fusion to dCas9. Additionally, we find that there are optimal targets within the murine Tissue Inhibitor of Metalloproteinase 2 (TIMP2) promoter that results in maximal transcriptional activation and that multiple sgRNAs expressed simultaneously results in a synergistic enhancement of promoter activity. Finally, using a genome‐wide bioinformatics approach we identified a number of potential off target sites for a given sgRNA‐target within the promoter of other genes. Selective activation of TIMP2 with the CRISPR‐Cas9 system opens up a strategy for treating diseases associated with over activity of the Matrix Metalloproteinases.Grant Funding Source: Supported by the Training Program in Translational Cardiovascular Science, 5 T32 HL 7936‐12
Temporal transcriptomic changes in canine liver slices compared to native liver (1148.3)
Authors: Liguori Michael , Klapczynski Marcin , Kowalkowski Kenneth , Ditewig Amy , Blomme Eric , Buck Wayne , Yang Yi 
Published: June 2021

Precision cut tissue slices retain the multicellular, structural, and functional features of the original organ and therefore offer a potentially more relevant approach to interrogate toxicity and pharmacology. However, limited data are available regarding transcriptomic changes occurring after slicing and in culture. Here, global mRNA profiles of canine liver slices immediately after preparation (0 h) and up to 48 h in culture were evaluated relative to whole liver. Liver slices from a male beagle dog were prepared and cultured at 37°C in an O2 rich atmosphere for 0, 24 and 48 h. Integrity of the slices was monitored using routine viability markers. Total RNA was isolated for analysis on Affymetrix canine genome microarrays. At 0 h, only minor perturbation of the transcriptome was apparent, indicating similarity to liver at the transcriptional level. After 24 to 48 h, ~8300 expression changes were observed. At 0 h, mRNA levels of phase I/II drug metabolizing enzymes were unchanged, but thereafter declined. Apoptotic signaling and proteasome mRNAs increased after 24 h, but not at 0 h, suggesting minor early stress signals. After 24 h, fibrosis pathway activation was evident with substantial increases in IL‐6 and IL‐8 mRNA. Unlike primary hepatocytes, a test substance can be incubated in slices at 0 h, allowing for the retention of an organ‐like molecular environment and thus offering advantages compared to monolayers.Grant Funding Source: Supported by AbbVie
Influence of beta‐sitosterol on ER and gene PS2 expression in breast cancer cell line T47D cells (1148.7)
Authors: Tao Shiying , Niu Jianzhao , Zhao Piwen , Hao Qinxiu , Yang Meijuan , Sun Yanling , Zhang Lijuan , Wang Jifeng 
Published: June 2021

Objective: To observe the effect of beta‐sitosterol on ER and downstream gene PS2 expression in breast cancer cell line T47D cells, and explore its phytoestrogenic mechanism. Methods: mRNA expressions of ER‐alpha,ER‐beta and its downstream gene PS2 in T47D were assessed by RT ‐PCR, protein expressions of ER‐alpha,ER‐beta in T47D were assessed by western‐blot. ICI182 780 was employed to observe the influence on the gene expressions of PS2 and protein expressions of ER‐alpha,ER‐beta. Results: beta‐sitosterol up‐regulated ER‐alpha,ER‐beta mRNA expression.PS2 gene are induced in response to beta‐sitosterol treatment. The effect on PS2 gene expression was inhibited by pure estrogen receptor antagonist ICI182780. Beta‐sitosterol increased the level of T47D cells ER‐alpha, ER‐beta proteins expression, respectively. The effect was completely antagonized by ICI182 780. Conclusion: beta‐sitosterol up‐regulated gene PS2 expression through ER. And beta‐sitosterol possessed phytoestrogenic effect by activating ER. Supporttrd by 111 project (B07007)and NSFC(30772849,81273887).Grant Funding Source: Suooorted by 111 projict(b07007) and NSFC（30772849,81273887）
Myocardial force and strain comparisons in control and MLP deficient mice (1150.1)
Authors: Michaelides Marcos , Georgiadou Soteria , Constantinides Christakis 
Published: June 2021

No prior study has directly characterized or quantified in‐vivo strain and force/stress responses from the left ventricular (LV) epicardium. This study quantifies and compares in‐vivo epicardial force and strain in control and transgenic hearts using micro sensors.Male mice (N=39) [C57BL/6 (n=26), 129/Sv (n=5), wild‐type (WT) C57×129Sv (n=5), and MLP (LIM‐protein knockout) (n=3)] were studied under 1.5% isoflurane. Piezoelectric crystals were placed at longitudinal (L) epicardial loci at basal (B), middle (M) and apical (A) LV regions, with the independent and/or concurrent placement of a force sensor. Longitudinal contractile and relaxation strains ranged between 4.8‐9.3 % in (B/M/A) regions of C57BL/6 mice and in mid‐LV regions of 129/Sv, WT, and MLP mice. Measured forces ranged between 3.1‐8.9 mN. Measurements were also conducted in normal mice following afterload‐challenges. Concurrent B‐L force‐displacement recordings in C57BL/6 mice yielded stress‐strain responses exhibiting bi‐modal active force contractile characteristics. Middle‐LV forces were significantly lower in MLP mice compared to WT controls (5.9±0.4 vs. 8.9±0.2 mN, p<0.0001), evidence of a defective excitation‐contraction coupling in the dilated, fibrotic, and stiffer myocardium.In‐vivo force and strain quantification is demonstrated in normal and trangenic murine hearts with potential usefulness to cardiac disease.Grant Funding Source: Supported by the Research Promotion Foundation
Matrix metalloproteinase‐12 inhibition causes cardiac dysfunction post‐myocardial infarction in mice (1151.3)
Authors: Iyer Rugmani , Patterson Nicolle , Dive Vincent , Lindsey Merry 
Published: June 2021

Matrix Metalloproteinases ‐12 (MMP‐12) deficiency diminishes the capacity to degrade extracellular matrix, but whether MMP‐12 inhibition post‐myocardial infarction (MI) is an effective therapeutic strategy has not been studied. Male C57BL/6J mice (3‐6 months old, n=60) were subjected to left coronary artery ligation. Saline or MMP‐12 inhibitor (MMP‐12i; 0.5 mg/kg/day) were delivered 3h post‐MI, and the mice were sacrificed at days 0 (no MI), 1 and 7 post‐MI. Infarct area was similar in the saline and MMP‐12i (p=0.70) groups, indicating similar initial injury. Survival rates were 33% for saline and 50% for the MMP‐12i (p=0.29). MMP‐12i group had 33±1% reduction in MMP‐12 activity at d1 post‐MI compared to the saline group (p<0.05). MMP‐12 inhibition worsened LV function at d7 post‐MI, as shown by larger LV volumes and decreased ejection fraction (all p<0.05). CD44, a cell adhesion molecule was 50±1% lower in RNA and protein levels in MMP12i group (both p<0.05). Hyaluronan (HA), a ligand of CD44, had reduced clearance at d1 and d7 post‐MI in MMP12i group. Accumulated HA increased levels of an apoptosis marker, caspase 3 at d7 post‐MI in the MMP12i group. Overexpression of caspase 3 leads to attenuated cardiac function post‐MI. In conclusion, MMP‐12 inhibition post‐MI reduced CD44‐HA interaction leading to increased apoptosis which resulted in LV dysfunction.Grant Funding Source: Supported by HHSN268201000036C, HL051971, R01 HL‐075360, 5I01BX000505
Blockade of platelet‐derived growth factor on cardiac fibrogenic response (1152.10)
Authors: Liu Chang , Zhao Wenyuan , Zhao Tieqiang , Meng Weixin , Chen Yuanjian , Liu Hongyu , Sun Yao 
Published: June 2021

Interstitial fibrosis appears in the viable myocardium of the infarcted heart (MI) and contributes to ventricular dysfunction. Platelet‐derived growth factor (PDGF) is a family of growth factors that stimulates cell growth, differentiation and migration. Our previous study has shown that PDGF‐D and PDGF receptors (PDGFR) were increased in the viable myocardium of the infarcted heart and cells expressing PDGF/PDGFR were predominantly fibroblasts. Herein, we sought to determine whether PDGF is involved in the development of interstitial fibrosis in the infarcted heart. MI was induced by the left coronary artery ligation in rats with or without receiving PDGF receptor antagonist (imatinib, 40mg/kg/d). Sham‐operated rats served as controls. We detected the effect of PDGF blockade on cardiac collagen synthesis and degradation at one week postMI. We found that compared to controls, TGF‐β1, type I collagen, TIMP‐1, TIMP‐2, MMP‐2 and MMP‐9 expressions were significantly increased in the viable myocardium of the left ventricle. PDGFR blockade significantly attenuated TGF‐β1, type I collagen, TIMP‐1/‐2 and MMP‐2 expressions compared to MI rats. These observations indicate that PDGF plays a role in the development of interstitial fibrosis in the viable myocardium of the infarcted heart. Our findings suggest the potential therapeutic effect of PDGFR blockade on interstitial fibrosis in the infarcted heart.Grant Funding Source: NIH
Ethanol‐induced cardiac fibrosis is mediated by NADPH oxidase (1152.12)
Authors: Mouton Alan , Walker Morgan , El Hajj Milad , Molina Patricia , Gilpin Nicholas , Gardner Jason 
Published: June 2021

Chronic ethanol (EtOH) abuse leads to cardiomyopathy that is characterized by extensive cardiac fibrosis and dilatation. We hypothesize that EtOH causes cardiac fibrosis by fibroblast activation via NADPH oxidase (NOX)‐induced oxidative stress. Male Sprague‐Dawley rats were intermittently exposed to EtOH vapor for 2 wks (14hr ON/10hr OFF; peak blood alcohol levels of ~200 mg/dL). Time‐matched controls were exposed to room air. Fixed mid‐LV sections stained with picrosirius red were assessed for collagen volume fraction (CVF). To examine the direct effects of EtOH on fibroblasts, isolated adult rat cardiac fibroblasts were treated with EtOH (50mM) for 24 hr, and cell‐conditioned media assayed for hydroxyproline (HPro). Protein expression was determined by Western blot; NOX activity was assessed by NADPH consumption assay; and cellular superoxide production was determined using DHE. Collagen gel contraction assay was used to assess fibroblast activity. In vivo EtOH‐exposure significantly increased LV CVF (1.6% vs 0.9% for control) and NOX‐2 and ‐4 expression (157.0% and 185.6% of control). EtOH significantly increased cardiac fibroblast secretion of collagen (HPro: 1.09±0.09 vs 0.75±0.07 µg/ml for control), and this was prevented by NOX inhibition (DPI; 10µM). EtOH increased fibroblast expression of α‐SMA (150.0% of control) and NOX‐2 (307.2%), but not NOX‐4. EtOH also significantly increased NOX activity (0.39±0.02 vs 0.27±0.00 nmol/mg/min) and superoxide production. EtOH increased gel contraction by fibroblasts (65.4% vs 42.4% control), which was also prevented by NOX inhibition. In conclusion, chronic intermittent EtOH vapor exposure produced cardiac fibrosis, which our data indicate is the result of a NOX‐dependent increase in fibroblast activity and collagen secretion.Grant Funding Source: Supported by an LSUHSC ADACE Pilot Grant (jdg).
Scleraxis regulates myocardial extracellular matrix synthesis and homeostasis (1152.14)
Authors: Bagchi Rushita , Roche Patricia , Aroutiounova Nina , Schweitzer Ronen , Czubryt Michael 
Published: June 2021

The cardiac extracellular matrix (ECM) functions as a source of mechanical and biochemical cues for cellular differentiation and growth in addition to its role as a physical scaffold for cellular constituents. The mechanisms governing basal ECM production are incompletely understood. Our laboratory has previously demonstrated the role of scleraxis (Scx) as a required regulator of human collagen Iα2 gene expression. Here we investigate the consequences of loss of Scx on matrix composition and turnover. We hypothesized that Scx functions to broadly regulate cardiac ECM production. DNA microarray analyses of Scx null (KO) murine hearts revealed marked changes in ECM components occurring due to loss of Scx. Histological assessment of Scx KO hearts revealed a marked reduction in abundance of collagen fibrils and overall fibrotic components as compared to wildtype (WT) hearts. Immunoblotting and qPCR analyses of these hearts confirmed a significant downregulation of fibrillar collagen types I, III and V. shRNA‐ mediated knockdown of Scx further refined the role of Scx as a direct regulator of these collagen genes. The expression of proteoglycan components including fibromodulin and lumican were reduced in Scx KO myocardium. Matrix metalloproteinases (MMPs), a prime regulator of matrix degradation and homeostasis, were also significantly altered in their expression particularly MMP2 and MMP9 as demonstrated by gelatin zymography and qPCR analyses. Thus Scx may be a direct modulator of the activity of these remodeling enzymes. The widespread effects of scleraxis deletion supports a central role for scleraxis in governing the normal expression and structure of the cardiac ECM.Grant Funding Source: Supported by the Canadian Institutes of Health Research and St. Boniface Research Foundation
Inhibition of Na/K‐ATPase signaling regulates cardiac remodeling induced by partial nephrectomy in mice (1152.3)
Authors: Drummond Christopher , Liu Jiang , Cooper Christopher , Xie Zijian , Shapiro Joseph , Tian Jiang 
Published: June 2021

Previous studies found that induction of renal dysfunction by 5/6th partial nephrectomy (PNx) resulted in cardiac hypertrophy and increased systolic blood pressure in rats and mice. Using Na/K‐ATPase (NKA) α1 heterozygous mice (α1+/‐) and their wild‐type (WT) littermates, we further demonstrate that PNx surgery or MBG infusion leads to increased cardiac cell death and decreased cell survival signaling in cardiac tissues compared to WT animals. To study the role of NKA signaling, we employed a newly developed Src inhibiting peptide (pNaKtide) derived from NKA. Both α1+/‐ and WT animals were subjected to PNx or sham surgeries. A 25mg/kg dose of pNaKtide was administered biweekly at the 12th, 14th, and 16th week after PNx surgery. One week after the first injection of pNaKtide, systolic blood pressure decreases significantly in α1+/‐ mice and slightly in WT animals. Significant decreases in diastolic dimension are also observed in both WT and α1+/‐ animals given PNx. In addition, both WT and α1+/‐ mice saw significant reversals of PNx induced cardiac hypertrophy indicated by heart weight/body weight ratio. This reversal was more significant in α1+/‐ vs WT animals. These data highlight the importance of Na/K‐ATPase signaling in cardiovascular remodeling.Grant Funding Source: Supported by NIH grants R01 HL‐105649 and HL‐109015
Cardioprotective effects of prior transient ACE inhibition ő role of cardiac fibroblast (1152.7)
Authors: Hale Taben , Biwer Lauren , Ramaiah Priya , Shahid Wafa , D'Souza Karen 
Published: June 2021

Transient ACE inhibition induces persistent changes that protect against future L‐NAME‐induced cardiac fibrosis and inflammation. This study investigates whether these observations can be mechanistically linked to changes in cardiac fibroblasts.We measured cardiac levels of chemokines, macrophages, and proliferating cells following 7 days of L‐NAME (L) in SHR that were previously treated with placebo (C+L) or enalapril (E+L) for 2wks followed by 2wk washout. Cardiac fibroblasts were isolated and cultured to passage 1.L‐NAME induced microinfarcts and vascular injury in both groups. In C+L, but not E+L there was an increase in macrophage‐recruiting chemokines. By day 7 of L‐NAME there was a marked increase in cardiac macrophages and proliferating cells in C+L, but not E+L. In vitro, fibroblasts from C+L were hyperproliferative, demonstrated increased Collagen I gene expression, and increased MCP‐1 production.These findings demonstrate that L‐NAME produces phenotypic changes in fibroblasts that persist in vitro that suggest that fibroblasts may participate in macrophage recruitment in addition to collagen production. However, fibroblasts isolated from SHR previously treated with enalapril display a different phenotype that more closely resembles cells from control hearts. ACE inhibitor‐induced cardioprotection may result, in part, from a modification of the cardiac fibroblast population.Grant Funding Source: Supported by: Sarver Heart Foundation, Steven M Gootter Foundation