bluuebunny/crossref_metadata_2025_split huggingface dataset:
EFFECT OF HYPERBARIC OXYGEN ON THE RAT CARDIOMYOCYTE AFTER 6 HOURS OF AN ACUTE MYOCARDIAL INFARCTION
Authors: Gutierrez‐Martinez Luis Alberto , Juan‐Dominguez Vania Don , Kormanovski‐Kovzova Alexandre , Castillo‐Hernandez Maria Carmen , Guevara‐Balcazar Gustavo 
Published: June 2021

The current treatment of myocardial infarct (MI) is thrombolysis, or ideally percutaneous coronary angioplasty. Alternatively, benefits of hyperbaric oxygen (HBO) have been reported during an MI in progress, where it seems to reduce the number of infarcted cardiomyocytes favoring their neovascularization.Material and methodsMI was induced by ligation of the anterior descending coronary artery. HBO was administered by a hyperbaric chamber 100%, at 2 atmospheres of pressure, for one hour daily for 15 days. After this time the rats were sacrificed, the size of the infarcted area was measured histologically, and the number of vascular buttons and neoformation vessels was determined by detecting the Vascular Endothelial Growth Factor (VGEFR2) and CD34 by immunohistochemistry, respectively. Three groups of 10 rats each were compared: group A (HBO without MI), group B (MI with HBO), and group C (MI without HBO).ResultsImmunohistochemically, a greater number of VEGFR2 positive vascular buttons were found in the rats of group C than those of group B (7.15 vs 2.55 per field, p = 0.01), but in the group B there are more CD34 positive neoformation vessels (7.8 vs 4.2 per field, p = 0.01).ConclusionsHBO promotes early neovascularization of the rat cardiomyocyte after 6 hours of an MI.Support or Funding InformationSIP, COFAA, IPNThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Cardiac Troponin‐T in Cardiac H9c2 Cells During Co‐Culture with Mesenchymal Stem Cells
Authors: Boomsma Robert A. , Jousma Jordan 
Published: June 2021

Previous studies have shown that intravenous injection of mouse bone marrow mesenchymal stem cells (MSC) attenuate the loss of function in the heart after coronary artery occlusion (Boomsma et al, Int J Cardiol 122:17 2007) and secrete paracrine factors that may be responsible for their beneficial effect (Boomsma and Geenen, PLoSONE 7:e3685, 2012). Mouse MSC were cultured in Mesencult + 20% serum supplement (Stem Cell Technologies) and rat cardiac H9c2 cells were cultured in DMEM + 10% FBS until confluent. MSC and H9c2 cells were then combined equally and plated in 6‐well dishes at a total concentration of 1.0 ×106 cells/well in DMEM + 10% FBS. After 2 days the media was changed to DMEM + 1% heat inactivated horse serum to promote myocyte differentiation. Control cultures containing H9c2 cells alone at the same cellular concentrations were treated similarly. After 1, 3, 5, and 7 days of culture in 1% serum RNA was isolated using RNAqueous 4 PCR (Ambion) and treated with DNAse. Cardiac troponin‐T (cTnT) was quantified in H9c2 cells by RT‐qPCR using an iScript cDNA Synthesis Kit and iQ SYBR Green Mastermix (BioRad). Relative expression was determined using the reference gene GAPDH; no‐RT and water controls were included. Species specific primers for rat cTnT and rat GAPDH were designed in order to separate out expression of cTnT in H9c2 cells (rat) from MSC (mouse). Mean and standard error was calculated for each group and significance was determined using Student's t‐test (p<0.05, n=5–6). Relative expression of cTnT in H9c2 cells was low in both co‐culture (0.422 ± 0.157) or alone (0.427 ± 0.065) after 1 day in 1% serum, but increased more than 10‐fold after 3–7 days in both co‐culture or alone. cTnT expression in H9c2 cells was significantly greater after 5 days in co‐culture with MSC (4.204 ± 0.524) compared to H9c2 cells cultured for 5 days alone (2.353 ± 0.510; p<0.05). Expression in H9c2 cells cultured alone continued to increase after 7 days (5.958 ± 1.179) such that there was no statistical difference from cells in co‐culture (3.918 ± 1.880). This study demonstrates that MSC accelerate cTnT expression in H9c2 cells and may partially explain the beneficial effect of MSC on cardiac function in vivo.Support or Funding InformationSupported by a VanderVelde Research Scholarship and a Summer Research Grant from Trinity Christian College.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Direct Effects of High Sodium on Histone Deacetylases in RAW264.7 Macrophages
Authors: Molina Patrick Axel , De Miguel Carmen , Dunaway Luke , Pollock Jennifer S. 
Published: June 2021

Evidence demonstrates a key role of macrophage (MΦ) activation in salt‐sensitive hypertension with pro‐inflammatory (M1) phenotype transition following high salt (NaCl) exposure. Our lab has shown that high Na+ regulates histone deacetylase 1 (HDAC1) expression in the kidney and HDAC9 was previously shown to regulate the M1 phenotype. However, it is unknown whether high Na specifically regulates MΦ HDAC1 or HDAC9 expression. The MΦ cell line RAW264.7 was cultured in DMEM (10% FBS) to confluency and exposed to 40mM NaCl, sodium acetate, choline chloride or mannitol for 24 hours to determine whether the gene expression is dependent on high Na or osmolality. To determine whether this in vitro system modulates the M1 or profibrotic M2 phenotype, we determined expression of interleukin 1β (IL‐1β), interleukin 12α (IL‐12α), and nitric oxide synthase 2 (NOS2) in response to 100ng/mL lipopolysaccharide (LPS) or 10ng/mL interleukin‐4 (IL‐4). Three independent experiments were completed. Data are presented as relative expression fold difference and one‐way ANOVA utilized for statistical analysis with significance at p<0.05. LPS exposure leads to M1 MΦ phenotype with increased IL‐1β and NOS2 (1713±618 and 42±6, respectively). Traditionally, IL‐4 is thought to induce the anti‐inflammatory M2 phenotype but 24hr treatment did not demonstrate a difference in IL‐12α from control, IL‐4 or LPS treatment, supporting the requirement of additional M2‐specific stimuli in parallel with IL‐4. Additionally, to examine an effect of high salt on LPS or IL‐4, MΦ were also exposed to IL‐1β + NaCl or IL‐4 + NaCl. There were no changes in HDAC1 mRNA expression following NaCl, sodium acetate, choline chloride, mannitol, LPS, LPS+NaCl, and IL‐4. IL‐4+NaCl decreased HDAC1 expression by 59.3±3.5%. There was no change in HDAC9 mRNA expression following any treatment group. These studies suggest that high sodium does not induce HDAC1 or HDAC9 gene expression in cultured macrophages. Further studies are warranted to determine protein abundance as well as in vivo effects of high salt diets, and in particular examination of changes in resident kidney macrophages.Support or Funding InformationNIH P01 HL69999, NIH P01 HL136267, AHA SFRN, MSTP T32 GM008361This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Maresin‐1 Activates Resolution Sensor and Improves Left Ventricle Function In Acute Heart Failure
Authors: Halade Ganesh V. , Travis Roman , Kain Vasundhara , Jadapalli Jeevan Kumar 
Published: June 2021

Acute inflammatory response is an active process coincides with resolving phase in myocardial infarction (MI), which enables the leukocyte directed clearance, if failed, lead to non‐resolving inflammation. Leukocyte, particularly macrophages are known to biosynthesize maresin‐1 (MaR1) post‐MI. However, the role of exogenous MaR1 is unclear in heart failure (HF) settings. Young 8–12 week‐old C57BL/6J male mice were subjected to coronary ligation and were treated with MaR1 (4μg/kg; subcutaneous) 3 hours post‐MI for day 1 or until day 5. Post‐MI saline‐injected mice served as MI‐control and no‐MI mice as d0 naïve controls. MaR1‐injected mice reduced LV and lung mass‐to‐body weight ratios compare to MI‐controls indicating decreased edematous hypertrophy and lung edema post‐MI d5. Comprehensive heart function analysis indicate reduced end‐diastolic volume (EDD) and end systolic volume (ESD) in MaR1‐injected mice thereby improved fractional shortening and global longitudinal strain compared to MI‐control at d5 post‐MI. MaR1 activated N‐formyl peptide receptor 2 (FPR2; p<0.05) expression in LV infarct compared to MI‐control group. Leukocyte trafficking analysis revealed that MaR1‐injected mice expedited neutrophil clearance (CD11b+/F4/80−/Ly6G+; 1.2±0.4%) from the infarcted heart compared with MI‐control (4.1±0.3%). Post‐MI, MaR1‐injected mice increased proresolving CD11b+/F480+/Ly6Clo macrophages at post‐MI day 5. MaR1 injected mice displayed higher levels of pro‐resolving markers Mrc‐1 (3.6±.5 fold; p<0.05) and Arg‐1 (5.2±1 fold; p<0.05) at day 1 and day 5 post‐MI compared with MI‐control. MaR1 showed multifactorial benefits by reducing cardiorenal inflammation and reduced NGAL and increased nephrin expression compared to MI‐control is an indicative of limited cardiorenal pathology post‐MI. In conclusion, MaR1 activated resolution sensor FPR2 to facilitate proresolving actions, thereby improved cardiac function and attenuated cardiorenal inflammation in HF pathophysiology.Support or Funding InformationHL132989 to GVH, 16POST31000008 to VKThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ATP‐Citrate Lyase is an Epigenetic Regulator to Promote Nephropathy in Obesity and Type 2 Diabetes
Authors: Li Yan Chun , Chen Yinyin , Deb Dilip K. 
Published: June 2021

Obesity and type 2 diabetes are leading causes of chronic kidney disease, but the mechanism whereby obesity and diabetes promote renal injury remains poorly understood. Hyperglycemia, hyperlipidemia and chronic inflammation are key features of obesity and type 2 diabetes. We hypothesize that hyperglycemia and hyperlipidemia drive excess nutrient flows into kidney cells and alter cellular metabolism. As a result, increased intracellular acetyl‐CoA provides the substrate for lipid biosynthesis as well as for histone acetylation in the genome. We used ob/ob BTBR mice, an obese model of susceptible diabetic nephropathy, to test this hypothesis. The ob/ob mice developed age‐dependent hyperglycemia and albuminuria. Histology revealed marked glomerular expansion, glomerulosclerosis and ectopic lipid accumulation in the kidney compared with ob/+ mice. Total lipids, triglyceride and cholesterol contents in the renal cortex were markedly elevated. The expression of key enzymes involved in fatty acid and cholesterol biosynthesis (acetyl‐CoA carboxylase (ACC), fatty acid synthase (FAS), HMG‐CoA reductase (HMGCR)) and pro‐fibrotic factors (TGF‐b1, fibronectin (FN)) was also markedly increased in the cortex, accompanied by a dramatic up‐regulation of ATP‐citrate lyase (Acly), an enzyme that converts citrate to acetyl‐CoA. Histone acetylation at H3K9/14 and H3K27, but not H3K18, was also dramatically elevated in the cortex. Inhibition of Acly activity by SB‐204990 not only blocked histone acetylation but also suppressed the expression of TGF‐b1, FN, ACC, FAS and HMGCR in ob/ob kidneys. ChIP assays confirmed that the promoters of all these genes were hyperacetylated at the H3K9/14 and H3K27 sites. We further used mesangial cell (MC) cultures to study the roles of Acly in renal fibrogenesis and ectopic lipogenesis. To mimic the condition of hyperglycemia, hyperlipidemia and chronic inflammation, we exposed MCs to high glucose (HG), sodium palmitate (P) and TNF‐a. A combination of HG, P and TNF‐a dramatically stimulated Acly expression and enzymatic activity, increased H3K4/19 and H3K27 acetylation, up‐regulated TGF‐b1, FN, ACC, FAS and HMGCR expression and promoted histone acetylation in these gene promoters. Blockage of Acly by SB‐204990 or siRNA attenuated these regulatory processes, whereas overexpression of Acly enhanced these regulations. The stimulation of Acly expression by HG, P and TNF‐a was mediated by CREB and NF‐kB at Acly gene promoter. These data strongly suggest that Acly is a critical epigenetic regulator to promote renal injury in obesity and type 2 diabetes.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic Flux Analysis Reveals Effects of Fatty Acid Metabolism on the Urea Cycle in Diabetic Kidney Disease
Authors: Jagannathan Narendra Suhas , Ching Jianhong , Kovalik Jean‐Paul , Tucker‐Kellogg Lisa 
Published: June 2021

Diabetic Kidney Disease (DKD) is one of the most common and serious complications of Type II Diabetes Mellitus (T2DM). Previous work measured plasma metabolites in DKD patients versus T2DM controls without DKD. DKD patients showed elevated hydroxyl‐ and dicarboxyl‐acylcarnitines. These metabolites are produced by omega‐oxidation (ωOx), a microsomal process that is known to be upregulated during mitochondrial dysregulation and inborn errors of metabolism. Briefly, dicarboxylic fatty acids from ωOx can be converted to dicarboxyl‐acylcarnitines or transported to the peroxisome for breakdown through peroxisomal β‐oxidation. ωOx is a poorly‐studied pathway that has never been subjected to computational simulations, nor included in detail in genome‐scale computational models of human metabolism.We developed a computational model of ωOx and integrated it with a previously published metabolic flux model of interplay between three tissue types (liver, muscle, adipose) in diabetes. We then performed metabolic flux simulations on this extended model using Flux‐Balance Analysis (FBA). Monte‐Carlo Flux sampling techniques identified cellular reactions with flux levels that were differentially regulated by allowing/blocking ωOx, or by allowing/blocking efflux of peroxisomal beta‐oxidation intermediates. Forcing the model to utilize ωOx caused consistent flux changes in nitrogen processing and the urea cycle (for muscle and liver), suggesting altered levels of urea and ammonia in plasma. While other pathways were also affected to different degrees, the effect was less consistent than with the urea cycle.In conclusion, we provide the first metabolic model that incorporates ωOx and peroxisomal beta‐oxidation in a multi‐tissue setting. Simulating a model of diabetes with ωOx identified links between mitochondrial dysfunction, fatty acid processing and nitrogen metabolism. Flux sampling simulations are useful for generating experimental hypotheses and for shedding light on systems‐level effects, but experimental validation is required before inferring causality. The importance of urea as an osmolyte and metabolite motivates further study of this relationship, which could have important implications for renal function.Support or Funding Information1. National Research Foundation (NRF), Prime Minister's Office, Singapore, under its CREATE programme, Singapore‐MIT Alliance for Research and Technology (SMART) BioSystems and Micromechanics (BioSyM) IRG2. Duke‐NUS SRP Phase 2 Research Block GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High‐resolution optical imaging of synchronization in the renal circulation.
Authors: Postnov Dmitry , Marsh Donald J. , Holstein‐Rathlou Niels‐Henrik , Cupples Will , Sosnovtseva Olga 
Published: June 2021

Study of nephron to nephron interaction and of the corresponding signaling pathways is fundamental for understanding how the kidney operates and why it malfunctions. Micropuncture studies have shown that nephrons signal to each other over short distances and can synchronize their activity accordingly. Because micropuncture can be applied to no more than 2 or 3 nephrons simultaneously, however, the technique is incapable of estimating the size of the synchronized field. In our earlier studies, we used a commercial laser speckle imaging camera to measure blood flow dynamics over the renal surface, but limitations of the technology available at the time restricted spatiotemporal resolution, making it impossible to capture signals from efferent arterioles of single nephrons at reasonable signal‐to‐noise ratio.To resolve signals from separate nephrons in a sufficiently large field of view we developed a new methodology for renal laser speckle imaging. Our setup provides spatial resolution down to 0.8□m per pixel and imaging frequency up to 160Hz.We applied the new technology to record ~1.5x1.5 mm2 sections of the renal surface in control and under IV stimuli, resolving 30–70 “star” vessels per section. We analyzed spectral properties of recorded signals in time and studied inter‐nephron magnitude coherence as well as frequency and phase locking. The analysis showed the dynamic evolution of clusters of various sizes, including the formation of large (>30 vessels) long‐lived TGF frequency locked clusters (>10 periods). To our knowledge, it is the first study to estimate of the number of nephrons in the superficial synchronized field and to show the evolution of synchronous clusters in time.Support or Funding InformationSupported by Novo Nordisk Foundation. Project number NNF17OC0025224.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Wnt signaling regulates macula densa structure and function
Authors: Shroff Urvi Nikhil , Gyarmati Georgina , Izuhara Audrey , Riquier‐Brison Anne , Peti‐Peterdi Janos 
Published: June 2021

The macula densa (MD), a plaque of 15–20 specialized cells, sits at the vascular pole of the glomerulus and plays a critical role in maintaining renal blood flow and glomerular filtration rate via tubuloglomerular feedback. Changes in renin state of the juxtaglomerular apparatus are known to be associated with alterations in MD cell morphology, although the underlying mechanism and importance of this phenomenon have been elusive. The canonical Wnt‐β catenin signaling pathway is known to regulate several cellular processes including embryonic development, cell proliferation, differentiation and homoeostasis. Glycogen synthase kinase 3 beta (GSK3b), a serine/threonine kinase, is involved in the phosphorylation and subsequent destablization of β catenin; thus, GSK3b plays a critical role in regulating the Wnt‐β catenin pathway. Lithium is a commonly used inhibitor of GSK3b in the kidney. Given the importance of Wnt‐β catenin in maintaining epithelial cell structure, the present studies aimed to explore its role in regulating MD morphology and function. Wild‐type mice were treated with lithium chloride for 2 weeks (40 mmol/kg chow for 7 days, and 60 mmol LiCl/kg chow subsequently), followed by harvesting and histological analysis of the kidneys. Lithium treatment caused a significant increase in the number of proliferating MD cells as quantified by Ki67 and EdU staining. As compared to control mice, MD cell number in the plaque was higher in lithium‐treated groups with several hyperplastic MD plaques observed containing > 40 cells. MD nucleus‐to‐cell volume ratio was 30% higher after lithium treatment. Using a O‐propargyl‐puromycin incorporation‐based protein synthesis imaging assay, a 3‐fold increase in protein synthesis was observed in MD cells after lithium treatment. Identical MD cell morphological alterations were observed in the newly generated nNOS‐Ctnnb1(Ex3) mice, a Wnt gain‐of‐function genetic model, confirming MD cell Wnt specificity of lithium treatment. In summary, Wnt signaling is an important regulator of MD morphology and cell biology, which may be involved in traditional renin and hemodynamic and novel tissue remodeling functions of MD cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rho Kinase Inhibitors Are Involved in L‐Type Voltage‐dependent Calcium Channel Pathway in Vascular Smooth Muscle Cells
Authors: Guan Zhengrong , Baty Josh , Zhang Shali , Inscho Edward W. 
Published: June 2021

Voltage‐dependent L‐type calcium channels (L‐VDCC) and the rho/rho kinase pathway are two predominate intracellular pathways that regulate renal microvascular tone and reactivity. Traditionally, these two pathways have been thought to act independently; however, recent evidence suggests these pathways are convergent. We hypothesized that the rho kinase inhibitors influenced L‐VDCC signaling. The effects of rho kinase inhibitors Y‐27632 or RKI‐1447 on KCl‐depolarization were assessed in afferent arterioles using the in vitro blood‐perfused rat juxtamedullary nephron preparation. Superfusion of KCl (30, 60, and 90mM) led to concentration‐dependent vasoconstriction of afferent arterioles reducing the diameter to 95±6, 56±4 and 42±3% of the baseline, respectively (p<0.05). Addition of Y‐27632 (1, 5 and 10 μM) or RKI‐1447 (0.1, 1 and 10 μM) significantly increased starting diameter by 16–65% (n=6/each, p<0.05). The KCl‐induced vasoconstriction with 1 μM Y‐27632 or 0.1 and 1 μM RKI‐1447 was similar to control, but markedly attenuated with 5 and 10 μM Y‐27632, and 10 μM RKI‐1447 (p<0.05 vs KCl alone). The afferent arteriole diameter averaged 99±2, 83±2, 71±4%, and 98±0.8, 89±5, 81±6% of the baseline for 5 and 10 μM Y‐27632, and 97±2, 89±3 and 88±4% of the baseline for 10 μM RKI‐1447, respectively (n=6/each, p<0.05 vs KCl alone). Changes in intracellular calcium concentration [Ca2+]i were estimated by fura‐2‐AM fluorescence (ratio F340/380) during KCl‐depolarization in cultured A7r5 cells (n=46–118/each group) and in freshly isolated renal microvascular smooth muscle cells (RMVSMCs, n=32–118/each group). Administration of 90 mM KCl increased fura‐2 fluorescence markedly in both cell types. The KCl‐induced increase of the F340/F380 ratio was unaltered by 0.1 or 0.5 μM Y‐27632 but was significantly attenuated by Y‐27632 between 1–10 μM by 50–56% (p<0.05 vs KCl alone) in A7r5 cells but only at 10 μM in RMVSMCs by 42% (p<0.05 vs KCl alone). RKI‐1447 however, significantly attenuated KCl‐mediated elevation of [Ca2+]i in both cell types regardless of the concentrations applied (0.1–10 μM, p<0.05), indicating that Y‐27632 and RKI‐1447 decrease L‐VDCC function. These studies indicate that the rho kinase inhibitors, Y‐27632 and RKI‐1447 partially inhibit L‐VDCC function and participate in L‐VDCC signaling.Support or Funding InformationThis study was supported by a Grant‐in‐Aid (15GRNT25240015) from the American Heart Association Great Southeast Affiliate and a grant from NIH (DK106500) to ZG, and by grants from NIH (DK044628, HL074167, HL098135 and HL095499) to EWI.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.