bluuebunny/crossref_metadata_2025_split huggingface dataset:
Endothelial cell‐specific knockout of epithelial sodium channel prevents vascular stiffening in response to a western diet in female mice
Authors: Aroor Annayya , Jia Guanghong , Manrique Camila , Lastra Guido , Barron Brady J , Dongqing Chen , Sun Zhe , Meininger Gerald A , Whaley‐Connell Adam T , Jaisser Frederic , Sowers James R 
Published: November 2023

Arterial stiffening, a strong and an independent predictor of future cardiovascular events, has emerged as a therapeutic target for reduction of cardiovascular disease. Over‐nutrition/obesity predisposes individuals, particularly women, to arterial stiffening. We have recently developed a clinically relevant murine model of high fat and high fructose diet (western diet, WD) feeding which exhibits arterial stiffening associated with vascular and cardiac dysfunction. In this model, female mice have high plasma aldosterone levels and WD feeding increases mineralocorticoid receptor (MR) expression in the vasculature. One of the mechanisms by which MR activation promotes endothelial stiffness is through increased expression and activation of epithelial sodium channel (EnNaC) in endothelial cells (ECs). In this regard, we showed WD feeding was associated with increased expression of epithelial sodium channel in ECs (EnNaC) and prevention of WD induced aortic EC stiffness and impaired relaxation of the aorta by the administration of a very low dose of the EnNAC inhibitor, amiloride. In this study, we tested the hypotheses that specific deletion of EnNaC, decreases aortic EC stiffness and improves vascular relaxation in WD‐fed female mice. To produce cell specific deletion of the EnNaC gene,”floxed” EnNaC mice were serially crossed with Tie 2‐Cre transgene mice. This resulted in marked suppression of EnNaC expression in ECs. Female KO mice and littermate controls were fed a WD with excess fat (46%) and fructose (17.5%) for 12 weeks. Compared to mice fed a control diet (CD), aortic EC stiffness, measured ex vivo by AFM was significantly increased in WD‐fed mice and this was prevented in EnNaC KO mice fed a WD. The decrease in EC stiffness was associated with improvement in endothelial dependent aortic relaxation with acetylcholine. Taken together, these findings support the notion that a WD promotes ECMR activation of EnNaC in ECs and associated vascular stiffness which is a marker/predictor of cardiovascular disease.Support or Funding InformationNIH (R01 HL73101‐01A and R01 HL107910‐01) and the Veterans Affairs Merit System (I01 BX001981) to JRS. K08HL129074‐01 to CM.
Diabetes Exacerbates Coronary Atherosclerosis in Ossabaw Miniature Swine with Metabolic Syndrome
Authors: Stivers Ben , Badin Jill , Kole Ayeeshik , Progar Victor , Pareddy Anisha , Alloosh Mouhamad , Sturek Michael 
Published: November 2023

Over 35% of all US adults are estimated to have metabolic syndrome (MetS), which is defined as the presence of three or more of the following risk factors: obesity, hypertension, glucose intolerance, insulin resistance, and dyslipidemia. Furthermore, the incidence of type 2 diabetes is rapidly growing, often in an overlapping population. MetS and diabetes both substantially increase the risk of coronary artery disease (CAD), which continues to be the leading cause of death in the US. We investigated the extent to which diabetic hyperglycemia superimposed on MetS exacerbates CAD. We performed this study in the well‐characterized Ossabaw miniature swine model of MetS and CAD. The beta‐cell toxin alloxan was administered to swine by intravenous injection, then swine were placed in two groups: pigs that responded only transiently to alloxan thereafter having fasting normoglycemia (MetS, n=8) and those that responded to alloxan becoming permanently diabetic (MetS/D, n=8). After alloxan administration, swine were fed an atherogenic diet for six months to induce the MetS phenotype. At sacrifice, we used intravascular ultrasound (IVUS) imaging to quantify the extent of CAD by assessment of percent plaque burden and spotty calcification in the left anterior descending artery. Percent plaque burden quantifies the percentage of the area within the external elastic membrane (EEM) that is occupied by atherosclerotic plaque, using the formula (EEM area – lumen area)/EEM area*100%. The percent plaque burden was significantly greater in left anterior descending artery of the MetS/D swine compared to MetS swine (12.9% vs. 3.7%; p<0.05). Furthermore, this difference was more prominent in the proximal 15 mm of the artery (17.1% vs. 3.9%; p<0.05). We assessed coronary artery calcification by identifying echogenic signal stronger than adventitia and with acoustic shadowing peripheral to the calcium deposit. We calculated a calcium index for each artery using the formula (Frames with calcification/Total frames analyzed)*(Maximal arc angle of calcium/360°). The calcium index was significantly greater in the proximal 15 mm of the left anterior descending artery of MetS/D swine compared to MetS swine (0.0204 vs. 0.00003; p<0.05). These results show significant positive correlation with Von Kossa stained sections from the proximal 15 mm of each vessel and analyzed for calcified area (r=0.84). Thus, we conclude that CAD, as measured by plaque burden and presence of calcification by IVUS, is exacerbated when diabetes is superimposed on MetS.Support or Funding InformationThis work was supported by NIH T35 HL110854, P30 DK097512, the Fortune‐Fry Ultrasound Research Fund, and Eli Lilly & Co.
Effects of Cellular Dehydration on Glucose Regulation in Healthy Males ‐ A Pilot Study
Authors: Jansen Lisa T , Adams JD , Johnson Evan C , Kavouras Stavros A 
Published: November 2023

Preliminary data from experimental animals have linked dehydration to glucose dysregulation via the action of vasopressin. Also epidemiological data suggest that humans with low water intake or elevated vasopressin are more prone to develop diabetes.PURPOSETherefore, the purpose of this study was to investigate the effects of cellular dehydration on glucose regulation in healthy males.METHODSFour non‐diabetic males (28.4 y±1, HbA1c 5.6±0.2%, BMI 23.6±1.2kg·m−2) were recruited for this preliminary study. Each subject underwent two experimental trials, consisting of a 2 h intravenous infusion of saline (ISO 0.9% and HYP 3.0% of NaCl, 0.1ml·kg·min−1 infusion rate), followed by a 2h oral glucose tolerance test. Blood samples were taken from an antecubital intravenous catheter in 30min intervals.RESULTSMean plasma osmolality was raised to 300±3mmol/kg for the HYP trial, while ISO maintained at 286± 2mmol/kg. Glycemic and insulin responses post glucose loading seemed to be delayed but spiked during HYP (150.6mg/dl glucose at min 60) while displaying a normal response during ISO (136mg/dl at min 60) trials. Homeostatic model assessment index of insulin resistance (HOMA IR) was HYP 0.93±0.24 vs. ISO: 0.85± 0.59, while Matsuda index of insulin sensitivity was HYP: 12.4±5.9 vs. ISO: 12.7±6.5. HYP seemed to cause increased insulin resistance and decreased insulin sensitivity compared to ISO.CONCLUSIONThe present data suggest that cellular dehydration induced by hypertonic saline infusion might be linked to glucose dysregulation in normal healthy adults.
Adipocyte‐specific deletion of HuR reduces diet‐induced obesity
Authors: Anthony Sarah R , Slone Samuel , Hui David Y , Owens Albert Phillip , Tranter Michael 
Published: November 2023

Obesity is classified by the World Health Organization as one of eight principle causes of preventable chronic disease, and is directly associated with decreased life expectancy due to increased risk of type II diabetes, chronic respiratory disease, stroke, peripheral vascular disease, cancer, and cardiovascular disease. As a serious threat to human health, research into novel pharmaceutical targets and therapies to combat obesity is of growing interest to the scientific community.Although the RNA‐binding protein HuR (Human antigen R) is highly expressed in adipocytes, its role in adipocyte function and adipocity is largely unknown. To conclusively determine the mechanistic role of HuR in adipocyte function, our laboratory has developed an adipocyte‐specific HuR knockout mouse (Adipo‐HuR−⁄−). These mice weigh less than wild‐type mice at baseline and have a reduced lean/fat body mass ratio. In addition, when placed on a chronic (22‐week) high fat diet (HFD, 60% kcal/fat), Adipo‐HuR−⁄− mice gain less weight compared to their wild‐type counterparts and display a decreased cold tolerance. Ongoing efforts to identify an underlying mechanism of action have shown no difference between wild‐type and Adipo‐HuR−⁄− mice following chronic HFD with regard to glucose tolerance or ratio of lean/fat body mass, but our data indicates a loss of UCP1 in the brown fat of Adipo‐HuR−⁄− mice.Support or Funding InformationThis work was partially funded by a University of Cincinnati Heart, Lung and Vascular Institute Near Horizons Collaborative Grant (MT, APO).
GENDER DIFFERENCES INVOLVING SEROTONIN (5‐HT) RECEPTORS IN TYPE I DIABETIC RATS
Authors: Moldovan Tudor , Cervantes Marisa , Bucan Jonathan , Walker Zachary , LePaige Bernard , Bhaskaran Subha , Banes‐Berceli Amy 
Published: November 2023

Diabetes mellitus is an important topic of research. In both Type I and Type II diabetes vascular and renal dysfunction are some of the many negative impacts observed in diabetic patients. Increased plasma levels of 5‐HT exist in male diabetic rodent models and previous data in male Japanese diabetic patients demonstrated that inhibition of 5‐HT2A receptors reduced proteinuria observed. Whether these are differences in males and females in 5‐HT levels and function is unknown. We hypothesized that increased levels 5‐HT receptors may be the cause of the vascular damage observed in diabetics; elevated levels of 5‐HT and 5‐HT receptors may lead to increased vasoconstriction. We used male and female Sprague‐Dawley rats (300–325g) and made them diabetic with Streptozotocin (STZ). At 14 days and 28 days post‐onset of diabetes we euthanized the animals, harvested tissues and blood vessels for Western blot and myograph analysis. At day 14, no significant differences in contractile responses in the blood vessels from either the male or female control vs diabetic rats in the thoracic aorta, renal, superior mesenteric, and femoral arteries existed. There was a leftward shift in the response curve to 5‐HT in the vessels from the males of both groups compared to the females. At day 28, there was an increased contractile response to 5‐HT in the aorta from the diabetic rats compared to control from male and females. The females also showed increased contraction to 5‐HT in the superior mesenteric artery at 28 days. There was no change observed in the renal artery or femoral artery from either sex at 28 days. We observed at days 14 and 28 in the aorta of the male rats that there was no differences in 5‐HT2A or 5‐HT2B receptor levels. In the kidney cortex at day 14 we observed increased expression of both 5‐HT2A and 5‐HT2B receptor levels. At day 28 only the 5‐HT2B receptor in the kidney cortex from the diabetic rats was increased. There was no difference in the 5‐HT2A receptor levels. In the aorta from the 14 day female rats we saw greater expression of the 5‐HT2A receptor in the diabetic rats compared to control and less expression of the 5‐HT2B receptor in the aorta from the diabetic rats compared to control. At 28 days we observed no difference between groups in the 5‐HT2A receptor levels but a small increase in the levels of the 5‐HT2B receptor in the aortas from the diabetic rats. In the cortex from the female rats we saw increased levels of the 5‐HT2A receptor at both 14 and 28 days and increased levels of the 5‐HT2B receptor levels at 28 days. These data clearly show altered expression of 5‐HT receptor during the development of diabetes and clear sex differences as well.Support or Funding InformationOakland University Center for Biomedical Research
A Novel Role for AT1R: Ang‐(1‐7)/Mas Signal Transduction
Authors: Exner Eric C , Stodola Timothy J , Hoffmann Brian R , Greene Andrew S 
Published: November 2023

Microvascular dysfunction is a key component of cardiovascular disease associated with many risk factors, including hypertension and diabetes. In models of renin suppression and hypertension, we have observed that microvascular function can be rescued by infusion of low doses of angiotensin‐(1‐7) (Ang‐(1‐7)). In previous studies, we detected the angiotensin II receptor type 1 (AT1R) as a potential interacting mechanism for transduction of signals caused by the binding of Ang‐(1‐7) to the Mas receptor. While the oligomerization of AT1R and other G‐protein coupled receptors has been described, the various partner receptors and physiological roles are just beginning to be illuminated. Here, we investigate whether AT1R is necessary for the effects of Mas mediated Ang‐(1‐7) signaling on microvascular function in a model of renin suppression and salt‐sensitive hypertension. We also seek to identify downstream mediators of Ang‐(1‐7)/Mas signaling that are disrupted in the absence of AT1R. In an AT1R knockout rat, the ability of Ang‐(1‐7) to rescue microvascular function in a model of hind‐limb angiogenesis was lost. In addition, Ang‐(1‐7) mediated enhancement of endothelial cell tube formation was compromised in AT1R knockout cells in vitro. AT1R dependent downstream mediators of Ang‐(1‐7)/Mas signaling were identified by combining high throughput, mass spectrometry based proteomics with gene expression data. These findings, combined with our previous work showing the presence of AT1R in the Ang‐(1‐7)/Mas signaling complex, suggest that physical proximity of Mas and AT1R is important for signal transduction. Further studies will validate targets identified here and investigate the role of Ang‐(1‐7) and the Mas/AT1R complex in regulation of endothelial progenitor cell function. These results suggest that oligomerization of receptors within the RAS may be of clinical importance, as regulation of microvascular function via the RAS is both pathophysiologically and pharmacologically relevant.Support or Funding InformationThis work was supported by NHLBI grants HL082798, T32HL7852‐18 and F30HL131153 as well as the Kern Foundation Innovation Award granted to Dr. Andrew S. Greene.
Zinc‐finger nuclease knockout of dual‐specificity protein phosphatase‐5 protects against DOCA‐salt induced chronic renal injury
Authors: Fan Fan , Murphy Sydney R , Ge Ying , Mims Paige N , Maeda Kenji J , Williams Jan M , Roman Richard J 
Published: November 2023

We recently reported that zinc‐finger nuclease knockout (KO) of dual‐specificity protein phosphatase‐5 (Dusp5) enhances the myogenic response and autoregulation of cerebral blood flow in FHH.1BN rats. We identified 4 single nucleotide polymorphisms in Dusp5 in FHH as compared with BN rats, two of which alter CpG sites and another one causes G155R mutation. In the present study, we evaluated whether Dusp5 also plays a role in the regulation of autoregulation of renal blood flow and renal function in a Dusp5 knockout (KO) rat in the FHH.1BN genetic background, which an 11 bp frame‐shift deletion is introduced by zinc‐finger nuclease and a premature stop codon at AA 121 of Dusp5 protein is produced. The expression of Dusp5 was lower in FHH and in Dusp5 KO rats in comparison to the FHH.1BN rats. The level of phosphorylated ERK1/2 (p‐ERK1/2), a substrate of Dusp5, was enhanced in the KO rats. The blood pressure and urinary protein excretion was similar in Dusp5 KO (n = 43) and FHH.1BN rats (n = 30) when they were at 12‐week of age (114 ± 2 vs. 115 ± 1 mmHg and 32 ± 2 vs. 29 ± 3 mg/day). In response to an elevation in pressure from 60 to 120 mm Hg, the renal afferent arterioles (Af‐arts) constricted by 29 ± 2% and 11 ± 1%, and the vascular tone was increased to 36 ± 2% vs 19 ± 1%, respectively in Dusp5 KO (n = 6) in comparison to FHH.1BN rats (n = 17). Blood pressure was 169 ± 5 vs. 177 ± 5 mmHg in Dusp5 KO (n = 11) vs. FHH.1BN rats (n = 14) after 3‐week DOCA‐salt treatment. However, proteinuria was significantly reduced in Dusp5 KO rats (258 ± 22 mg/day, n = 12) in comparison to FHH.1BN rats (338 ± 30 mg/day, n = 12) with the same treatment. The renal injury induced by DOCA‐salt was markedly reduced in Dusp5 KO vs. FHH.1BN rats as the glomerular injury scores were 2.49 ± 0.03 vs. 2.54 ± 0.01; the areas of renal fibrosis were 5.5 ± 0.5% vs. 8.3 ± 0.3%; the areas of renal protein casts were 0.5 ± 0.2% vs. 2.4 ± 1.0% and renal arteriolar media‐to‐lumen ratios were 0.64 ± 0.03 vs. 0.81 ± 0.04, respectively. These results indicate that Dusp5 modulates myogenic reactivity in the renal circulation and protects against DOCA‐salt hypertension induced chronic renal injury. It supports the view that upregulation of the expression of Dusp5 and/or an activating mutation in Dusp5 may contribute to the impaired myogenic response and autoregulation of renal blood flow and promote the development of chronic renal injury in FHH rats.Support or Funding InformationThis study was supported by grants HL36279 (RJR) and DK104184 (RJR), AG050049 (FF), P20GM104357 (RJR, FF and JMW) from the National Institutes of Health; 16GRNT31200036 (FF) and 12SDG9440034 (JMW) from the American Heart Association. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
ERalpha Protects Against Lipoprotein‐Associated Oxidative Stress to Regulate Vascular Tone in Both Male and Female Mice
Authors: Kim Seong Chul , Lee Jean‐Pyo , Hamblin Milton H. 
Published: November 2023

Lectin‐like oxidized low‐density lipoprotein receptor‐1 (LOX‐1) is a major vascular scavenger receptor for binding and cellular uptake of oxidized low‐density lipoproteins (oxLDL). LOX‐1 is an important mediator of oxLDL‐stimulated pathways that trigger vascular dysfunction, inflammation, and oxidative stress. However, the mechanisms protecting against lipoprotein‐related oxidative stress to maintain normal vascular tone are understudied. Here we investigated the connection of ERalpha‐activated vascular protection with the oxidative status of vascular rings. We preincubated male and female aortic vessels with 10 μM ox‐LDL for 30 minutes, and after measuring phenylephrine contractile responses, treated the aortas with 10−12 to 10−5 μM of 4,4′,4″‐(4‐propyl‐[1H]pyrazole‐1,3,5‐triyl)tris‐phenol (PPT), a specific ERalpha agonist. We found that PPT induced relaxation in aortic rings preincubated with ox‐LDL in both sexes, demonstrating a vascular protective role of ERalpha in lipoprotein‐associated oxidative stress. Given the interplay between ERalpha signaling and lipoprotein‐associated oxidative stress, we evaluated the expression level of LOX‐1 in PPT‐stimulated mouse endothelial cells in vitro. Western blot analysis shows that LOX‐1 expression was attenuated in endothelial cells following treatment with PPT. Collectively, these data suggest a protective role of ERalpha against lipoprotein‐associated oxidative stress in regulating vascular tone. Moreover, we anticipate these studies will provide a broader look into intracellular signaling mechanisms that influence vascular tone and events.Support or Funding InformationSupported by 5K12HD043451
Cardiorespiratory Dysfunction in Advanced Lung Disease versus Systemic Lupus
Authors: Wooten Liana C , Chin Lisa M.K. , Collins John , Chan Leighton , Keyser Randall E. 
Published: November 2023

The cardiorespiratory responses to a peak exercise challenge were compared among women manifesting three autoimmune mediated conditions, pulmonary hypertension (PH), interstitial lung disease (ILD) and systemic lupus erythematous (SLE), and sedentary but otherwise healthy controls.Methods80 women (PH N=28, age=54.0 ± 10.3, BMI=31.8 ± 7.3; ILD N=9, age=55.8 ± 7.2, BMI=29.7 ± 4.6; SLE N=25, age=35.1 ± 9.0, BMI=27.6 ± 5.8; Control N=18, age=36.8 ± 7.3, BMI=39.3 ± 8.8) completed a treadmill cardiopulmonary exercise test (CPET) to volitional exhaustion. Heart rate was obtained by continuous EKG monitoring and pulmonary gas exchange information was obtained using a breath‐by‐breath cardiopulmonary exercise testing system. Peak exercise oxygen consumption (VO2), heart rate (HR) and respiratory exchange ratio (RER) were recorded as an average of the last 8 consecutive breaths. Oxygen pulse (O2‐pulse) was calculated and the V‐slope method was used to determine VO2 observed at the anaerobic threshold (AT‐VO2). Data were analyzed for statistical significance (p<0.05) using analysis of variance and Cohen's d‐statistic for effect size.ResultsPeak RER was ≥ 1.10 in all groups except PH (RER = 1.00 ± 0.25). Peak VO2 and AT‐VO2 was significantly lower in the PH, ILD, and SLE groups compared to controls (p<0.01).

Peak VO2(ml/kg/min)
AT‐VO2(ml/kg/min)
Peak HR(beats/min)
O2‐Pulse(ml/beat)

PH
15.3 ± 5.4
10.3 ± 4.3
139 ± 17
9.2 ± 4.6

ILD
18.2 ± 5.9
11.7 ± 3.8
150 ± 19
9.6 ± 4.6

SLE
19.4 ± 4.8
12.5 ± 3.2
171 ± 18
8.3 ± 1.9

Control
28.8 ± 5.2
16.4 ± 2.2
180 ± 10
11.0 ± 2.3

Peak VO2 was lower in PH than in SLE (p=0.031), whereas AT‐VO2 was similar among PH, ILD and SLE. No difference in peak HR was observed between SLE and controls or between PH and ILD, however peak HR was significantly higher in SLE than in ILD (p=0.009) and PH (p=0.001). While not statistically significant, the size of the group difference in O2‐pulse was large (Cohen's d = 1.283).DiscussionPrevious studies have demonstrated similarity in peak cardiac output and stroke volume between healthy controls and patient subsets with autoimmune conditions such as PH, ILD and SLE. The decline in peak VO2 and HR across the PH and ILD groups, compared to controls, in concomitance with the disappearance of this decline with respect to O2‐pulse may suggest peripheral muscle fatigue as a factor limiting exercise tolerance in these groups. The large effect in SLE with respect to the decline in O2‐pulse and similarity in peak HR with controls may provide further evidence for peripheral oxygen extraction impairment in women with SLE.Support or Funding InformationSupported by NIH/NICHD/NCMRR HD39775; NIH IRP 1 Z01 CL060068‐02 CC
Dysregulation of Vasodilatory Pathways is a Contributory Factor to Acute Chest Syndrome
Authors: Frazier Krystle , Young Sheila M , Cook Kayleigh , Ibe Basil Obijiaku 
Published: November 2023

Acute chest syndrome (ACS), the second most common cause of hospitalization for sickle cell disease (SCD) patients and cause for 25% of premature death cases, remains a therapeutic challenge due to the unknown nature of the syndrome's development. SCD patients exhibit a baseline leukocytosis, or pro‐inflammatory state, that increases their risk of developing ACS. Red blood cells (RBC) of SCD patients induce injury to the lung microvasculature resulting in the release of clotting factors and inflammatory mediators. Our hypothesis is that these mediators sustain pulmonary endothelial injury and RBC adhesion to the vessel wall resulting in pulmonary vascular occlusion and ultimately ACS.ObjectiveTo investigate a mechanism by which RBCs stimulate production and release of inflammatory cytokines and proteins in SCD.MethodsStudies were conducted on human pulmonary microvascular endothelial cells (HPMEC) with and without overlay with RBCs or neutrophils of SCD patients in the steady state. Inflammatory mediators were measured by ELISA or Western blotting. Data were analyzed statistically at p<0.05.ResultsHPMEC cultured with RBCs from SCD patients released 40% more leukotriene C4 and 6‐fold more 11β‐PGF2α and more TxA2 than controls. Release of prostacyclin by SCD RBC blood was not different from control.ConclusionsSickle cell mutation‐bearing erythrocytes cause release of inflammatory mediators that activate vasoconstrictive pathways in HPMEC and down‐regulate vasodilatory pathways. This condition may eventually overwhelm the ability of the HPMEC to secrete vasodilators. A closer look at the interaction of these mediators and microvascular endothelium may reveal genetic polymorphisms and provide a potential for therapeutic targeting with gene silencing technology.Support or Funding InformationAMA Foundation