bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Association between Suppressed Transformation of Fibroblasts to Myofibroblasts and Fibrolysis Induced by Copper Supplementation in Monkeys of Myocardial Ischemic Infarction
Authors: Xiao Ying , Liu Yinjie , Han Pengfei , Luo Qin , Wang Heng , Kang Y. James 
Published: June 2021

Myocardial ischemia is associated with cardiac tissue copper (Cu) loss and fibrosis. Cu supplementation diminishes cardiac fibrosis and improves cardiac function. The transformation of fibroblasts to myofibroblasts is highly responsible for the pathogenesis of cardiac fibrosis. This study was undertaken to test the hypothesis that suppression of the transformation from fibroblasts to myofibroblasts is responsible for Cu‐induced reduction of cardiac fibrosis. Rhesus monkeys of 4–5 years old were subjected to coronary artery ligation to induce myocardial infarction (MI). At the fourth week after the surgery, an ultrasound‐directed Cu‐albumin microbubble organ‐specific Cu delivery technique was used to treat myocardial infarct monkeys twice a week for four weeks. This treatment significantly increased Cu concentrations in the infarct area, reduced the infarct size, and loosened collagen cross‐linking network. Total fibroblasts labeled with vimentin were increased in the infarct area, and this increase was not changed by Cu supplementation. In contrast, myofibroblasts, labeled with both vimentin and α‐smooth muscle actin (a‐SMA), were also significantly increased in the infarct area, but Cu supplementation significantly reduced this increase, along with decreased mRNA levels of type I and III collagens. This decrease in collagen production was accompanied by a significant increase in metalloproteinase‐1 (MMP‐1) and the MMP‐1 producing fibroblasts (vimentin+ and MMP‐1+ cells). In addition, primary cultures of neonatal rat cardiac fibroblasts were treated with 0.1 μM angiotensin II (Ang II) to increase cellular α‐SMA concentrations, mimicking the transformation from fibroblasts to myofibroblasts. This increase in α‐SMA production was significantly suppressed by addition of 5 μM CuSO4 in the cultures. These results suggest that Cu inhibits the transformation of fibroblasts to myofibroblasts, leading to the reduction of collagen synthesis and the increase in collagen degradation, and their combination results in fibrolysis in the infarcted myocardium.Support or Funding InformationThis study was supported by National Science Foundation of China (81230004 and 81300109).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The NLRP3 mediates the recruitment of lymphocytes in the cavernosal tissue and erectile function impairment in DOCA/salt mice.
Authors: Fais Rafael Sobrano , Rodrigues Fernanda Luciano , da Silva Josiane Fernandes , Tostes Rita C. , Carneiro Fernando Silva 
Published: June 2021

IntroductionErectile dysfunction (ED) is associated with risk factors for the development of cardiovascular diseases such as hypertension. Increased activity of the immune system is closely linked to the development of hypertension and ED. Arterial hypertension is strongly influenced by pro‐inflammatory effects of NLRP3. NLRP3 activity promotes vascular changes, cardiac and renal hypertrophy and increase in blood pressure. In addition, NLRP3 is involved in the recruitment of cells of the immune system. Increasing evidence shows that increased lymphocyte infiltrate is involved in the genesis and/or aggravation of various cardiovascular diseases.HypothesisNLRP3 increases lymphocyte infiltrate, promotes impairment of corpora cavernosa (CC) reactivity and ED in DOCA/salt hypertensive mice.ObjectiveEvaluate the role of NLRP3 in lymphocyte recruitment in CC, impairment of erectile function and changes in CC reactivity in DOCA/salt mice.Methods(CEUA 213/2016). Male C57BL/6 (WT) and NLRP3 deficient mice (NLRP3−/−) were either treated with vehicle or induced to DOCA/salt hypertension. Fluorescence to FAM‐FLICA and histochemistry were performed to analyze respectively the caspase‐1 activity and the number of lymphocytes in CC. The intracavernous pressure (ICP) was measured. Reactivity of CC to acetylcholine (ACh) and sodium nitroprusside (SNP) was performed.ResultsDOCA/salt hypertension increased the caspase‐1 activity in the CC. In addition, we observed increased lymphocyte infiltrate in the CC (WTsham: 18.5 ± 1.0; WTdoca: 33.5 ± 1.4, n=6), impaired erectile function (decreased ICP in WTdoca in the frequencies of 4, 8, 12 and 16 Hz) and reduced relaxation for ACh (pD2 WTsham: 7.2 ± 0.08, n=7; WTdoca: 6.5 ± 0.12, n=7) and SNP (pD2 WTsham: 7.2 ± 0.07, n=6; WTdoca: 6.7 ± 0.06, n=7) in the CC of WT mice. The absence of NLRP3 prevented the increase in the caspase‐1 activity and in the lymphocyte infiltrate (NLRP3−/−sham: 12.0 ± 1.2; NLRP3−/−doca: 17.6 ± 1.9, n=5) in the CC. Furthermore, NLRP3 absence normalized the erectile function, the reactivity to ACh (pD2 NLRP3−/−sham: 7.0 ± 0,14, n=6; NLRP3−/−doca: 6.9 ± 0.16, n=7) and to SNP (pD2 NLRP3−/−sham: 8.2 ± 0.08, n=6; NLRP3−/−doca: 8.5 ± 0.07, n=7) in DOCA/salt hypertension.ConclusionNLRP3 mediates the lymphocyte recruitment, impairment in erectile function and CC reactivity, promoted by DOCA/salt hypertension.Support or Funding InformationFinancial support: FAPESP, CNPq, CAPES, CRID.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Aldosterone‐Induced Hypertension is T Lymphocyte‐Dependent and Attenuated by Activation of the G Protein‐Coupled Estrogen Receptor 1
Authors: Dinh Quynh Nhu , Vinh Antony , Kim Hyun Ah , Diep Henry , Broughton Bradley , Chrissobolis Sophocles , Drummond Grant , Sobey Christopher 
Published: June 2021

The G protein‐coupled estrogen receptor 1 (GPER), a membrane‐localised estrogen receptor, may modulate some of the effects of aldosterone. G‐1, a GPER agonist, can promote T cell‐mediated anti‐inflammatory actions and acutely lower blood pressure. This study aimed to test the effects of G‐1 and G‐15 (GPER antagonist) on aldosterone‐induced hypertension in male and female mice, and to examine the cellular mechanisms involved. C57Bl6 and RAG1‐deficient mice were treated with vehicle, aldosterone/salt (0.72 mg/kg/d SC plus 0.9% NaCl for drinking) ± G‐1 (0.03 mg/kg/d SC) and/or G‐15 (0.3 mg/kg/d SC) for 14 d. Some females were ovariectomised 7 d prior to treatment. T cells were adoptively transferred via IV injections 21 d prior to treatment. In males, aldosterone/salt caused a sustained increase in blood pressure of ~25 mmHg within 7 d and this increase was attenuated by ~50% in the presence of G‐1 (n=11–13, P<0.05). G‐15 alone did not alter hypertension caused by aldosterone/salt but it prevented the anti‐hypertensive effect of G‐1. By contrast, the pressor response to aldosterone/salt was delayed in females where an increase in blood pressure was only observed at 14 d; however co‐administration of aldosterone/salt with G‐15 resulted in a marked increase of ~20 mmHg from 7 d (n=6–8, P<0.05). Administration of aldosterone/salt produced a robust increase in blood pressure from d 7 in ovariectomised females (n=5–9, P<0.05), similar to that observed in males. T and B cells in kidneys of male and female C57Bl6 mice were found to express GPER. In male RAG1‐deficient mice, aldosterone/salt had virtually no effect on blood pressure (n=8, P<0.05). However, adoptive transfer of 7×106 T cells from male C57Bl6 into male RAG1‐deficient mice fully restored the pressor response to aldosterone/salt, which could be attenuated by co‐treatment with G‐1 (n=8–14, P<0.05). This study indicates that activation of GPER by G‐1 and/or endogenous estrogen profoundly attenuates aldosterone/salt‐induced hypertension in mice. T cells are key contributors to aldosterone/salt‐induced hypertension, and therapeutic activation of GPER on T cells may reduce hypertension caused by elevated levels of aldosterone.Support or Funding InformationNHMRC grantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of reactive oxygen species in gestational diabetes and the impact on the developing fetus
Authors: Arriaga Diana 
Published: June 2021

Gestational diabetes mellites (GDM) is characterized by elevated levels of glucose in a pregnant woman's bloodstream that occurs from an overproduction of hormones. Under GDM, elevated glucocorticoids (GC) and insulin like growth factor (IGF)‐1 hormones in the placenta may possibly lead to an increase in the amount of reactive oxygen species (ROS) in the embryo due to an increase in glucose utilization and mitochondrial activity. Overproduction of ROS can modify the genome, thus predisposing offspring's to diabetes and obesity later. The aim of the study is to investigate the mechanism by which GDM leads to an increased production of ROS and evaluate the epigenetic changes that occur. I hypothesize that under diabetic conditions, the embryo will have an increase in ROS production due to increased hormone levels, resulting in epigenetic changes in the offspring. An in vivo model for GDM will be established by feeding female mice a high‐fat diet for four weeks. Controls will be fed a standard diet. Fasting blood glucose levels and glucose tolerance tests will monitor the development of diabetes. Mating between a diabetic or control female will occur with a normal male. At 6.5 and 17.5 days of gestation and 7 days post birth, maternal, fetal and placental blood and tissue will be collected. Biochemical assays will evaluate tissue ROS levels and epigenetic changes. Blood analysis will measure hormone and glucose levels. Results from this study will hopefully lead to the development of treatments to lower the incidence of diabetes in future generations.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of extracellular vesicles released by hypoxic adipocytes in Insulin action
Authors: Mora Silvia , Mleczklo Justyna , Ortega Francisco , FernandezReal Jose Manuel , Falcon‐Perez Juan Manuel 
Published: June 2021

Extracellular vesicles (EVs) are released by many cells and provide a mechanism for intercellular communication in an autocrine and paracrine fashion. EVs are also released during pathological states. Adipocytes release EVs, however, the impact of these vesicles in glucose metabolism and insulin action has not been thoroughly investigated. We sought to investigate whether EVs released by metabolically challenged adipocytes exposed to hypoxia or to macrophage conditioned media affect insulin responsiveness and glucose transport. EVs were isolated from the culture supernatant of adipocytes cultured under normoxia, hypoxia (1% oxygen) or exposed to macrophage conditioned media (15%v/v). EVs were isolated from plasma of lean and obese individuals. EVs were used to treat cells and glucose transport and activation of insulin signalling cascades was measured. EVs released from hypoxic adipocytes but not those released by adipocytes exposed to macrophage conditioned media impaired insulin‐stimulated 2‐deoxyglucose uptake and reduced insulin‐stimulated Ser473 phosphorylation of AKT. ERK signalling was intact. Insulin stimulated glucose transport in muscle cells was also inhibited. EVs from obese individuals decreased insulin stimulated 2‐deoxyglucose uptake in adipocytes (p=0.0159). EVs from obese subjects contained distinct miRNA signatures. These findings indicate that EVs released by stressed adipocytes impair insulin action.Support or Funding InformationUniversity of Livepool, MINECO and FEDERThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inflammation and Kidney Injury in Diabetic African American Men
Authors: Cao Lei , Boston Ava M. , Jegede Olugbemiga Ebenezer , Harrison Scott Henry , Newman Robert Howard , Ongeri Elimelda Moige 
Published: June 2021

Chronic kidney disease is the most common complication of diabetes. Without interventions, diabetic kidney disease (DKD) progresses to end stage renal disease (ESRD). In the United States, minority ethnic groups (e.g. African Americans) are disproportionately burdened by DKD. While genetic factors contribute to the disparities in DKD, little is known about the underlying cellular and molecular mechanisms. Several studies have shown that inflammation plays a role in the pathophysiology of DKD and the fibrosis observed in diabetic nephropathy. The goal of the current study was to determine the association between specific inflammation markers and diabetic kidney injury in diabetic African American men, an underrepresented sub‐population with high risk for diabetes and DKD. To this end, diabetic patients with (n=21) and without (n=88) diagnosed kidney disease were recruited at the Cone Health Community Health and Wellness Center in Greensboro, NC. Age‐matched, non‐diabetic subjects (n=81) served as controls. Assays for albumin and creatinine were used for biochemical assessment of kidney function. The diabetic patients were then stratified into three groups based on their albumin to creatinine ratios (ACR); normoalbuminuria (ACR<30 mg/g), microalbuminuria (30 mg/g<ACR<300 mg/g), and macroalbuminuria (ACR>300 mg/g). Enzyme‐linked immunosorbent assays were used to measure the plasma and urinary levels of five inflammatory markers; two markers of system‐wide markers (adiponectin and C‐reactive protein (CRP)); and three markers involved in renal inflammation (tumor necrosis factor‐α (TNF‐α), interleukin‐6 (IL‐6), intercellular cell adhesion molecule‐1 (ICAM‐1)). Plasma levels of the pro‐inflammatory cytokines TNF‐α and IL‐6 were significantly higher in diabetic patients with micro‐ and macroalbuminuria when compared to non‐diabetic controls and diabetic patients with normoalbuminuria. Urinary levels for ICAM‐1 and the anti‐inflammatory cytokine, adiponectin, were higher in diabetic patients with macroalbuminuria than the other three groups. Plasma CRP was elevated in diabetic subjects compared to non‐diabetic controls, but did not correlate with the extent of diabetic kidney injury. In contrast, urinary ICAM‐1, plasma TNF‐α, and plasma adiponectin had moderate positive correlations with ACR. The plasma levels for ICAM‐1, IL‐6, and CRP were not significantly correlated with the ACR. The data suggest that certain inflammation pathways play a role in the pathogenesis of DKD in African American men.Support or Funding InformationThis study was supported by the Minority Men's Health Initiative (MMHI) through NIH/NIMHD Center Award # U54MD008621 (CFDA 93.307), Sub‐Awards # HU140400 (to RHN and EMO) and HU150006 (to EMO and SHH); and the NIH/NIGMS Award Number SC3GM102049 to EMO.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
nNOS in Embryonic Kidney Contributes to Glomerular Maturation
Authors: Der Balint , Gyarmati Georgina , Riquier‐Brison Anne , Shroff Urvi , Peti‐Peterdi Janos 
Published: June 2021

The regulatory role of neuronal nitric oxide synthase (nNOS) in renal and glomerular hemodynamics is well established. In addition, nNOS is involved in cellular migration in other organs, such as the brain. The highest level of nNOS expression occurs on day 6 after birth (P6), when the mouse kidney is still developing. The purpose of this study was to explore a novel, non‐traditional role of nNOS in renal development and glomerular maturation. To determine the timepoints and localization of nNOS expression in embryonic kidney, fluorescent reporter nNos+/−‐Cre+/+/mTmG+/+ mice were induced with tamoxifen at different timepoints during pregnancy. The specific inhibitor of nNOS, 7‐Nitroindazole (7‐NI) was administered daily by ip. injections (20 mg/kg) from E16.5 to P7 in pregnant Pod+/+‐Cre+/+/Confetti+/+ mice. Fetal kidneys were harvested after birth at P7 and further processed for histological analysis. During embryonic kidney development, nNOS expression is detectable in the medullary region as early as E16.5. At E19.5, nNOS is expressed in macula densa cells at the juxtaglomerular apparatus and in a few cells of the medullary collecting duct. At P3, nNOS was localized in the same cell types as in earlier stages. Pharmacological inhibition of nNOS for 10 days by daily 7‐NI treatment did not alter the kidney/body weight ratio compared to vehicle treatment, albeit the 7‐NI treated animal group had growth retardation (n=4 each). The diameter of glomeruli was 20% smaller, and the cumulative podocyte number in glomeruli was decreased by 40% in 7‐NI treated animals (n=4). Albeit 7‐NI treatment resulted in the reduction of body weight likely due to nonrenal effects, the embryonic presence of nNOS in macula densa cells and the negative impact of nNOS inhibition on glomerular diameter and podocyte number suggest a novel, non‐traditional role for nNOS in embryonic kidney development and glomerular maturation.Support or Funding InformationResearch support NIH NIDDK R01 DK64324This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glomerular Endothelial Cell Calcium Dynamics Visualized in vivo
Authors: Schiessl Ina Maria , Gyarmati Georgina , Peti‐Peterdi Janos 
Published: June 2021

Glomerular endothelial cells (GEnC) are a critically important component of the glomerular filtration barrier and are subject to continually oscillating hemodynamics‐based high mechanical forces. Several in vitro studies suggest a key role of calcium in endothelial cell functions, such as the production of nitric oxide and cell proliferation. However, little is known about GEnC calcium dynamics in vivo. Here, we generated a new genetic mouse model to study calcium levels in GEnCs during physiological and several pathophysiological conditions.Endothelial cell calcium reporter mice (Cdh5‐Cre × PC‐G5‐tdT) were developed by crossing Cdh5 (VeCadherin)‐CreERT2 and PC‐G5‐tdT mice. After tamoxifen‐induction, these mice co‐express tdTomato and the calcium sensitive fluorescent protein GCaMP5G in endothelial cells, which allows the study of intracellular calcium changes in a semi‐ratiometric fashion. Endothelial cell‐specificity of GCaMP5G expression in the kidney was confirmed by co‐localization of GCaMP5G with the endothelial marker CD31 using immunohistochemistry.Intravital 2‐photon imaging of fully induced Cdh5‐Cre × PC‐G5‐tdT kidneys at baseline revealed generally stable intracellular calcium levels in GEnCs, which were occasionally disrupted by spontaneous calcium elevations in single cells. Consistent with regular oscillations in myogenic contractions of the afferent arteriole, time‐lapse imaging of endothelial cells in the afferent arteriole depicted calcium fluctuations.Acute injection of sphingosine 1 phosphate (4 ug/g BW), which is an important factor of endothelial cell migration and proliferation, induced a 3‐fold increase of GEnC calcium levels. Furthermore, laser‐induced ablation of a single macula densa cell stimulated a pronounced calcium elevation in the afferent arteriole endothelium of the same nephron, suggesting a paracrine communication between macula densa and endothelial cells within the juxtaglomerular apparatus.We next analyzed GEnC calcium levels during pathologic alterations in glomerular hemodynamics. Acute bolus injection of L‐NAME (10 mg/kg BW) caused a 2‐fold increase of afferent endothelial cell calcium levels and increased GEnC calcium values 1.3‐fold. Laser‐induced formation of a local glomerular thrombus provoked a 1.7‐fold elevation of GCaMP5G/tdTomato ratio in affected GEnCs. Furthermore, GEnC calcium levels were assessed 1 week after unilateral ureteral obstruction (UUO), and found to be 2‐fold elevated compared to GEnCs in the contralateral kidneys.In summary, GEnC calcium levels fluctuate in response to physiological and pathophysiological hemodynamic changes. Intravital 2‐photon microscopy of newly generated Cdh5‐Cre × PC‐G5‐tdT kidneys is a promising new tool to improve our understanding of GEnC calcium dynamics in vivo and to further study its role in endothelial cell physiology and regeneration.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mutant Actn4 alters the distribution of intracellular stress within podocytes and contributes to their mechanical failure in response to stretch
Authors: Feng Di , Notbohm Jacob , He Shijie , Wang Minxian , Krishnan Ramaswamy , Pollak Martin 
Published: June 2021

Alpha‐actinin‐4 (ACTN4 in humans, Actn4 in mice) crosslinks actin filaments in podocytes, providing them mechanical resilience as they help maintain the glomerular filtration barrier. Although human disease‐causing mutant ACTN4 leads to brittle reconstituted actin networks in vitro, how this affects the ability of podocytes to withstand the external mechanical challenges within the glomerulus remains unknown. We studied primary podocytes isolated from wild‐type (WT) and disease‐causing mutant Actn4 knock‐in mice. We previously reported that, after transient stretch, WT cells recovered their baseline contraction and actin cytoskeleton breakages, whereas mutant podocytes developed irrecoverable reductions in their contraction and irreparable disruptions in their cytoskeletons. We now report that after stretch, when challenged in a centrifugal detachment assay, mutant podocytes (11/23) had a higher probability of detachment than WT (2/19) (odds ratio =7.42, 95% CI 1.27, 80.75; p=0.017). Through an analysis of baseline biophysical properties, we found that the actin filaments of mutant Actn4 podocytes displayed a more uniform distribution of orientation (aligned more parallel to each other) compared with the filaments of WT podocytes (which displayed more random orientation). We then computed baseline intracellular stress in terms of two constituent components: the magnitude of mean principal stress as tension (σ), and the orientation of first principal stress as principal orientation (θ). The spatial distribution of intracellular stress within the podocytes was measured using the autocorrelation function C(r). The distance over which the C(r) decays to a value of 0.5 was extrapolated as a singular value from the autocorrelation curves and is termed correlation length, or r1/2. The greater the r1/2 value, the greater the distance over which intracellular stress maintains uniformity. We found that mutant podocytes displayed a more uniform distribution of intracellular stress across the cell, both in terms of magnitude (r1/2 σ mean ± SE: mutant 18.0 ± 0.8 μm vs. WT 13.4 ± 0.8 μm) and orientation (r1/2 θ mean ± SE: mutant 34.7 ± 2.2 μm vs. WT 25.9 ± 2.3 μm). Moreover, the more uniform the intracellular stress was distributed at baseline, the less likely mutant podocytes recovered their baseline contraction after periodic stretch. Taken together, our findings provide novel insight into the mechanism by which ACTN4 mutation alters the alignment of the cytoskeleton and impairs the resiliency of the podocyte, leaving it vulnerable to mechanical stretch and more prone to detachment in its glomerular environment.Support or Funding InformationR37DK059588 and T32DK007199This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.