bluuebunny/crossref_metadata_2025_split huggingface dataset:
Exercise Mitigates Calpain 1 Induced Cardiac Dysfunction Through PCP4 in Diabetes
Authors: Chaturvedi Pankaj , Kunkel George H , Familtseva Anastasia , Kalani Anuradha , Tyagi Suresh C 
Published: November 2023

Calpain 1 is a ubiquitous calcium dependent cysteine protease and found in cytoplasm as well as mitochondria. We have reported earlier that active calpain 1 is translocated from cytosol to mitochondria and activates MMP9. Although studies have reported that activation of calpain 1 is deleterious to the heart in various aspects but there are no reports whether calpain 1 can degrade purkinje cell protein (PCP4) and affect cardiac rhythm in diabetes. Therefore, we hypothesize that calpain 1 degrades PCP4 in diabetes to cause cardiac dysfunction and exercise can mitigate this. To test this hypothesis we recruited four groups of mice 1)db/+ control, 2)db/+ control with exercise, 3)db/db, 4)db/db with exercise. The mice were exercised on treadmill at 7m/min for 300m for 8 weeks and in agreement with the American Veterinary Research Guidelines. HL‐1 atrial cell line for in vitro studies and isolated cardiomyocytes for ex‐vivo studies were used. To evaluate whether activation of calpain can degrade PCP4 we treated the isolated cardiomyocytes with high calcium (40mM) till they lose their shape and evaluated the expression of PCP4, calmodulin, calmodulin kinase and calpastatin with Western blots and real time PCR. The induction of high levels of calpain 1 significantly downregulated PCP4 along with upregulated levels of calmodulin and calmodulin kinase. We used the CRISPR/Cas9 plasmid to knock down calpain in HL‐1 myocytes which restored the levels of PCP4 along with calmodulin and calmodulin kinase. In vivo, we found upregulated levels of calpain 1 in db/db mice as compared to controls which were mitigated in the exercised mice. Exercise also mitigated the levels of PCP4, calmodulin and calmodulin kinase in db/db mice. Conclusively our data strongly suggests that in diabetes there is high induction of calpain 1 with degrades PCP4, a protein important for cardiac rhythm and exercise can mitigate arrhythmias.Support or Funding InformationAcknowledgements: This work was supported by NIH grants HL‐74185 and HL‐108621 and AHA grant 15POST23110021
Aging increases Reactive Oxygen Species and Interleukin‐17 concentrations in Peroxisome Proliferator Activated Receptor –alpha (PPAR‐α) knockout mice during Angiotensin II Hypertension through a NADPH Oxidase Subunit‐2 mechanism
Authors: Ananaba Ugoeze , Williams Nia C , Allard Joanne S , Lee Dexter L 
Published: November 2023

Aging decreases myocardial peroxisome proliferator activated receptor –alpha (PPAR‐α) expression. Activation of PPAR‐α reduces hypertension, cellular oxidative damage and inflammatory cytokine production. We hypothesize that the absence of PPAR‐α would increase mean arterial pressure (MAP) during Angiotensin (Ang II) hypertension in aged mice hearts by increasing the expression of NADPH (NOX) subunit‐2, reactive oxygen species and interleukin‐17 (IL‐17) production. Male (40 weeks old) PPAR‐α knockout (KO) mice and wild‐type (WT) mice were implanted with biotelemetry devices and infused with a slow pressor dose of Ang II (400 ng/kg/min) for 12 days. Separate groups of Ang II treated mice were given the PPAR‐α agonist, fenofibrate (Fen) (145 mg/kg/day). On day 12 of Ang II, MAP was increased in KO compared to WT, 163 ± 6 and 148 ± 3 mmHg, respectively. Heart expression of NOX‐2 was increased in Ang II‐treated KO (90 ± 10%) and WT (80 ± 8%) mice, when compared to no treatment controls. WT + Ang II + Fen NOX‐2 expression was decreased (27 ± 6%) when compared to WT + Ang II alone. TBARS concentration was increased in KO + Ang II (12.4 ± 1 mM), when compared to WT + Ang II (10 ± 1 mM). IL‐17 concentrations were increased in KO + Ang II (1.7 ± 0.5 ng/mL) and WT + Ang II (1.2 ± 0.2 ng/mL), when compared to their respective controls. Our results suggest increased myocardial NOX‐2 expression, TBARS and increased IL‐17 concentrations in 40 week old PPAR‐α KO mice are potential mechanisms for the increased Ang II‐induced blood pressure responses.Support or Funding Information#1R25AGO47843‐01
Acute effects of dietary nitrate supplementation on peripheral and central blood pressure in post‐menopausal women
Authors: Kim Danielle Jin‐Kwang , Barrett Megan A , Moore David J , Flanagan Michael P , Roe Carly , Kim‐Shapiro Daniel B , Proctor David N 
Published: November 2023

Dietary nitrate supplementation (in the form of beetroot juice) has been shown to have multiple cardiovascular health benefits, including both acute and chronic reductions in resting arterial blood pressure. The purpose of this study was to examine the effects of acute dietary nitrate supplementation on peripheral and central blood pressures in postmenopausal women, a population that exhibits accelerated vascular aging and cardiovascular disease progression. We hypothesized that 1) postmenopausal women would have higher resting central blood pressure compared to premenopausal women, and 2) acute dietary nitrate supplementation would reduce resting peripheral and central blood pressure in postmenopausal women. Eighteen women participated in this study (9 premenopausal women, 20–30 yrs; 9 postmenopausal women, 55–70 yrs). Supine peripheral blood pressure (Omron) and radial pulse wave derived central aortic pressure (SphygmoCor CvM, AtCor Medical) were measured after 10 minutes of quiet supine rest. A subset of the postmenopausal women also participated in a randomized double‐blind crossover study during which they consumed beetroot juice (BRJ; 140 mL Beet‐It Sport, James White Juice Company) or placebo (PLC; 140 mL nitrate‐depleted Beet‐It Sport, James White Juice Company) approximately 1 week apart. Supine peripheral and central blood pressures were measured prior to and 90 minutes following juice consumption.ResultsPostmenopausal women had significantly higher brachial and central blood pressures (systolic, diastolic, and mean), and augmentation index (AI) compared to premenopausal women (all p < 0.05). Acute nitrate supplementation resulted in greater reductions (p<0.05) in brachial mean arterial pressure, central mean arterial pressure, and central systolic blood pressure compared to those observed during the placebo visit. These findings suggest that acute dietary nitrate supplementation favorably modifies resting peripheral and central blood pressures in postmenopausal women.Support or Funding InformationPenn State Ambulatory Research Fund (JAFFE Endowment)
Knockout of TSP‐1 Prevents Hyperglycemia‐induced Atherosclerosis in STZ‐treated ApoE−/− Mice
Authors: Ganguly Rituparna , Sahu Soumyadip , Raman Priya 
Published: November 2023

Atherosclerosis, a macrovascular disease is responsible for ~70–80% long term deaths in diabetic patients. Several clinical studies and trials as well as recent animal studies have identified hyperglycemia, hallmark of Type 1 and Type 2 diabetes, as a critical risk factor for atherosclerotic complications associated with diabetes. Thrombospondin‐1 (TSP‐1) is a proatherogenic and anti‐angiogenic ‘matricellular’ protein genetically associated with development of atherosclerosis, coronary artery disease and myocardial infarction. TSP‐1 level is significantly increased in diabetic patients, diabetic animal models and glucose‐stimulated vascular cells (endothelial cells, vascular smooth muscle cells, fibroblasts) in vitro. Previous reports, including earlier evidence from our laboratory, suggest TSP‐1 as a potential link between hyperglycemia and accelerated atherogenesis in diabetics. We showed earlier that in STZ‐induced hyperglycemic ApoE−/− mice, the micronutrient chromium picolinate in vivo attenuates development of atherosclerotic lesions concomitant to downregulation of TSP1 expression in the aortic vessels. The goal of the current study was to investigate a direct role of TSP‐1 in hyperglycemia‐induced atherosclerosis. We generated TSP1−/−/ApoE−/− double knockout mice by crossing TSP1−/− mice with atherosclerotic ApoE−/− mice, both being in C57BL/6J background. Hyperglycemia (blood glucose>250mg/dl) was induced in six weeks old male ApoE−/− mice and TSP1−/−/ApoE−/− double knockout mice via intraperitoneal injection of 50 mg/kg/day of Streptozotocin (STZ) or sodium citrate buffer (vehicle) for 5 consecutive days. At endpoint, mice (18 wks. of age) were harvested for atherosclerotic lesion analyses. Specifically, Oil‐Red‐O and Hematoxylin‐eosin staining of the aortic root area showed a significant reduction in lipid laden atherosclerotic lesions (~2.4 fold) and intimal thickening (~32%) in STZ‐induced hyperglycemic TSP1−/−/ApoE−/− mice compared to STZ‐treated ApoE−/− mice. Characterization of lesion cellularity by immunostaining for PCNA (proliferating cell nuclear antigen) and alpha‐SMA (alpha smooth muscle actin) revealed reduction in PCNA staining (~33%) and vascular smooth muscle cell abundance (~24%) in hyperglycemic TSP1−/−/ApoE−/− mice compared to hyperglycemic ApoE−/− mice. Further, atherosclerotic lesions in the aortic sinus of STZ‐treated hyperglycemic TSP1−/−/ApoE−/− mice showed a significant reduction in both macrophage content and collagen accumulation (~68%) as compared to the STZ treated hyperglycemic ApoE−/− mice. Together these data clearly demonstrate that knockout of TSP‐1 in ApoE−/− mice protects against hyperglycemia‐induced atherosclerosis, suggesting TSP‐1 as a potential therapeutic target in diabetic macrovascular disease.Support or Funding InformationSupported by Diabetes Action Research and Education Foundation (DAREF) and NEOMED start‐up funds.
β‐cell Deletion of Nr4a1 and Nr4a3 Nuclear Receptors Impedes Mitochondrial Respiration and Insulin Secretion
Authors: Reynolds Merrick S , Hancock Chad R , Ray Jason D , Kener Kyle B , Hardman Jeremy M , Tessem Jeffery S 
Published: November 2023

Insulin secretion from β‐cells is dependent on the stimulation of mitochondrial respiration in response to an increase in glucose supply. Various studies have clearly demonstrated that the Nr4a family of orphan nuclear receptors are essential for fuel utilization and mitochondrial function in liver, muscle and fat. We have previously demonstrated that overexpression of Nr4a1 and Nr4a3 are sufficient to induce proliferation of pancreatic β‐cells. We examined the impact of nuclear receptors Nr4a1 and Nr4a3 on pancreatic β‐cell mitochondrial function and proliferation using lentivirus mediated shRNA knock‐down of Nr4a1 and Nr4a3 in INS‐1 derived 832/13 rat insulinoma cells. β‐cells deficient in Nr4a1 and Nr4a3 have a 37% reduction in proliferation rates (p<0.05). The reduction in β‐cell proliferation with Nr4a1 and Nr4a3 deficiency was accompanied by a significant decrease in glucose stimulated insulin secretion rates (GSIS) (26% decrease, p<0.05). GSIS was determined in cells preincubated in secretion assay buffer (SAB) for 1.5 hours containing 2.5 mM glucose followed by incubation in SAB with 16.7 mM glucose. High resolution respirometry (Oroboros O2k) was used to evaluate mitochondrial respiratory function and capacity with substrate‐uncoupler‐inhibitor titration protocols for both intact and permeabilized β‐cells. Permeabilized cells allow for an in depth assessment of chemiosmotic coupling within the β‐cells while the intact protocol evaluates a more physiologically relevant condition. We have shown that deletions of the Nr4a nuclear receptors clearly impact cellular respiration, lowering the oxygen consumption rate in all respiratory states of intact β‐cells between 40–65% and permeabilized β‐cells by 23–50% compared to GFP (p<0.05). Our results indicate that knockdown of Nr4a1 and Nr4a3 results in decreased expression Idh3a, Idh3g, Sdhb, and Mdh2, which would impede TCA cycle progression, and is consistent with the reduction observed in O2 consumption, ATP production and ultimately glucose stimulated insulin secretion. These data demonstrate for the first time that the Nr4a orphan nuclear receptor family members Nr4a1 and Nr4a3 are critical for basal β‐cell mitochondrial function and insulin secretion.Support or Funding InformationBrigham Young University
CARBONIC ANHYDRASE MEDIATES HEPATIC STEATOSIS AND VASCULAR STIFFNESS INDUCED BY A WESTERN DIET
Authors: Malcolm Threshia , Aroor Annayya , Bivin William , Sowers James , Woodman Ryan , Lockette Warren 
Published: November 2023

The typical “Western diet” (WD) is higher in fat and high fructose corn syrup when compared to a healthful diet (CD) that is lower in fat and does not contain high fructose corn syrup. A WD leads to insulin resistance and the metabolic syndrome that is characterized by fatty steatosis in the liver, a precursor of non‐alcoholic fatty liver disease, and vascular stiffness that presages atherosclerotic vascular disease and hypertension. Bicarbonate is a necessary precursor for fatty acid synthesis and the storage of depot fat in the liver and in perivascular tissue. We hypothesized that acetazolamide (ACT), a carbonic anhydrase inhibitor that blocks bicarbonate‐dependent formation of fatty acids and triglycerides, could prevent the fatty steatosis and vascular stiffness that is induced by a WD. Laboratory mice were fed a WD (46% kcals from fat; 36% kcals from carbohydrate, sucrose [17.5%] and high fructose corn syrup [17.5%]) or CD (10% kcals from fat; 72% kcals from carbohydrate; no high fructose corn syrup) for sixteen weeks with, and without, 2mM ACT in the drinking water ad libitum. Subsequently, rodents were weighed, and fat mass, lean mass, and body water was measured with magnetic resonance imaging (MRI). Hepatic steatosis was graded on a scale of 1–9 arbitrary units (A.U.) on hematoxylin and eosin stained liver tissue. Vascular stiffness was measured by atomic force microscopy (AFM) on aorta explants.

CD
WD
WD + ACT

Total Body Weight (values in g ± SEM)
23.1 ± 0.4
31.3 ± 1.1**
20.1 ± 1.5 (N.S.)

Total Fat Mass (values in g ± SEM)
4.7 ± 11.2
11.2 ± 1 **
5.1 ± 1.2 (N.S.)

Total Body Water/Total Body Weight (in % ± SEM)
65.1 ± 0.8
49.9 ± 2.0**
57.9 ± 2.6 (N.S.)

Hepatic Steatosis (in arbitrary units, AU, ± SEM)
3.75 ± 1.4
7.4 ± 0.5*
5.0 ± 1.1 (N.S.)

Vascular Stiffness (in kPa ± SEM)
4.72 ± 0.6
17.18 ± 3.2*
5.22 ± 1.0 (N.S.)

N = 20,
denotes P < 0.05,
denotes P < 0.01,
N.S. denotes not significantly different from control diet (CD).

Compared to a CD, sixteen weeks of a WD was associated with a gain in total body weight that was completely inhibited by ACT. The increase in total adipocyte fat mass and percentage of body weight from water in WD animals were also inhibited by ACT. Notably, inhibition of carbonic anhydrase resulted in a decrease in hepatic steatosis and aortic vascular stiffness. If excess adiposity is responsible for the untoward sequelae of the metabolic syndrome, chronic inhibition of carbonic anhydrase‐dependent formation of bicarbonate and fatty acid synthesis could result in the normalization of the metabolic profile associated with an unhealthy Western dietSupport or Funding InformationSupport for this project was provided by the University of Missouri.
Differential Effects of Toll‐like Receptors 2 and 4 in Endotoxin‐Induced Vascular Dysfunction
Authors: Battson Micah L , Lee Dustin M , Hou Shoufei , Jarrell Dillon K , Santangelo Kelly S , Cox‐York Kimberly A , Gentile Christopher L 
Published: November 2023

Bacterial endotoxin (lipopolysaccharide, LPS) may contribute to diabetes‐related inflammation and has been shown to cause generalized vascular dysfunction, including arterial hypocontractility and impaired endothelium‐dependent and –independent vasodilation. These vascular impairments are thought to be mediated in large part via Toll‐like receptor 4 (TLR4). However, contribution of other TLRs, in particular TLR2, has not been well characterized. We therefore examined the relative contribution of TLR2 and TLR4 in mediating the acute effects of LPS on vascular dysfunction. Vascular reactivity was assessed via ex vivo pressure myography in isolated carotid arteries from male wild‐type (WT), TLR2‐KO and TLR4‐KO mice following 1 hour incubation with or without endotoxin (E. coli LPS 50ug/ml). LPS decreased maximal constriction to the thromboxane A2 agonist U46619 in WT (CON: 30 ± 3% vs LPS: 17 ± 3%) but not TLR2‐KO (CON: 29 ± 4% vs LPS: 25 ± 3%) or TLR4‐KO (CON: 33 ± 2% vs LPS: 32 ± 6%). LPS reduced endothelium‐dependent dilation (EDD) to acetylcholine in WT (CON: 95 ± 1% vs LPS: 73 ± 4%) and TLR2‐KO (CON: 84 ± 3% vs LPS: 64 ± 5%) but not TLR4‐KO (CON: 94 ± 2% vs LPS: 89 ± 3%). Finally, LPS reduced maximal endothelium‐independent dilation (EID) to sodium nitroprusside in WT (CON: 94 ± 1% vs LPS: 76 ± 6%) but not TLR2 (CON: 78 ± 5% vs LPS: 74 ± 6%) or TLR4‐KO (CON: 89 ± 3% vs LPS: 80 ± 2%). These findings suggest that lack of TLR2 protects carotid arteries against the LPS‐induced reductions in constriction and EID and that arteries of TLR4‐KO mice display additional protection against impaired EDD. Future studies will investigate the mechanism underlying these differential effects and may provide insight into potential treatments for endotoxin‐related vascular dysfunction.Support or Funding InformationFunding: Boettcher Foundation's Webb‐Waring Biomedical Research Program (CLG); National Institutes of Health DK087777 (CLG)
Ufm1 E3 ligase Ufl1 is essential for adaptive hypertrophy and ER stress response in the heart
Authors: Li Jie , Yue Guihua , Ma Wenxia , Li Huizhong , Zhang Aizhen , Li Faqian , Li Honglin , Su Huabo 
Published: November 2023

Post‐translational modifications of proteins are important regulatory mechanisms of protein function. Protein modifications by Ufm1, a novel ubiquitin‐like protein, require Ufm1 E1 UBA5, E2 Ufc1 and E3 ligase Ufl1 and are emerging as a central theme in protein activity regulation. To date the significance of Ufm1 modification in health and disease is largely unknown. In this study, we reported that Ufl1 was down‐regulated in human failing hearts of ischemic cardiomyopathy and in the ischemic regions of mouse hearts subject to ischemia reperfusion injury. To investigate the effects of decreased Ufmylation in the heart, we generated mice with cardiomyocyte‐restricted deficiency of Ufl1 (Ufl1KO). Loss of Uf11 impaired Ufm1 conjugation and reduced the protein levels of its two known targets, RGDDK1 and C53, in mouse heart. The KO mice were viable and had no morphological abnormality at 2 months after birth. However, during ageing, Ufl1KO mice displayed reduced heart weight to body weight ratio, upregulation of cardiac fetal genes, increased apoptosis and increased fibrosis, which eventually led to left ventricular chamber dilatation, wall thinning and cardiac Huidysfunction at 6 months of age. When challenged by transverse aortic constriction (TAC)‐induced pressure overload, Ufl1KO mice quickly developed dilated cardiomyopathy and heart failure at 2 weeks post procedure without a prior phase of adaptive hypertrophy. RNA sequencing analysis revealed a significant alteration in the expression profile of ER stress‐related genes in Ufl1KO hearts, which was further confirmed by real‐time PCR analysis and western blots of several ER stress response factors. In primary cultured cardiomyocytes, knockdown of Ufl1 also blunted ER stress response and aggravated ER stress‐induced cell injury. Hence, our data demonstrate an essential role of Ufm1 modification in the heart under physiological and pathological conditions and reveal a critical role of Ufl1 in mediating adaptive ER stress response in cardiomyocytes.Support or Funding InformationThis work was supported in part by NIH grant R01HL124248 (to H.S.) and American Heart Association grant 11SDG6960011 (to H.S.).
The Effect of Monocrotaline‐Induced Heart Failure on Left‐ and Right‐Ventricular Function
Authors: Guild Sarah‐Jane , Han June‐Chiew , Power Amelia , Ward Marie‐Louise , Nisbet Linley A , Loiselle Denis S 
Published: November 2023

Monocrotaline is well established to cause pulmonary arterial hypertension and right heart failure. To characterize the development of monocrotaline‐induced heart failure in Wistar rats, Millar telemetry devices were used to follow arterial pressure and heart rate for six weeks following a single subcutaneous injection of either monocrotaline (60 mg/kg) or saline. Four weeks after monocrotaline injection, mean arterial, systolic, diastolic and pulse pressures all began to decrease progressively. This decrease in pressure was accompanied by a progressive increase in heart rate until 5 weeks post‐injection when heart rate began to fall again. Ventricular function was assessed 6 weeks post‐injection by placement of pressure volume catheters in both the left and right ventricles. Reduced ejection fraction and dP/dt max indicated reduced contractile function of both the left and right ventricles in the monocrotaline‐treated rats compared to the saline‐treated controls.Support or Funding InformationSupported by the National Heart Foundation of New Zealand
Longitudinal Study on Independent and Joint Effect of Brachial‐Ankle Pulse Wave Velocity and Blood Pressure Control on Incident Stroke in Hypertensive Patients
Authors: Song Yun , Xu Benjamin , Xu Richard , Tung Renee , Frank Eric , Tromble Wayne , Yang William , Zhang Yan , Li Jianping , Qin Xianhui , Wang Binyan , Hou Fan Fan , Huo Yong , Wang Hong 
Published: November 2023

BackgroundPulse wave velocity (PWV) has been shown to affect the effects of anti‐hypertensive treatment in prevention of cardiovascular diseases. Data are very limited regarding whether PWV is an independent predictor of stroke above and beyond hypertension control. In their current guidelines for the management of arterial hypertension, the European Society of Hypertension included PWV as a potential consideration in managing middle‐aged hypertensive patients with increased aortic stiffness. However, none of the current stroke prevention guidelines have considered PWV due to limited data.ObjectiveThis longitudinal analysis examined the independent and joint effect of brachial‐ankle PWV (baPWV) with hypertension control on the risk of first stroke.MethodsThis report included 3,310 hypertensive adults (age: 59.6 ± 7.5 years), a subset of the China Stroke Primary Prevention Trial (CSPPT) with baseline measurement for baPWV. All the participants had no prior history of stroke or myocardial infarction (MI).ResultsMean baseline baPWV was 18.76 (SD: 3.82) m/sec. During a median follow‐up of 4.5 years, 111 participants developed first stroke. The risk of stroke was higher among participants with baPWV in the highest quartile than that in the lower range (6.3% vs. 2.4%). Similarly, the participants with inadequate hypertension control had a higher risk of stroke than those with adequate control (5.1% vs. 1.8%). When baPWV and hypertension control were examined jointly, participants in the highest baPWV quartile and with inadequate control had the highest risk of stroke compared to their counterparts as demonstrated by Kaplan‐Meier curves (log rank test p<0.0001) and Cox proportional hazard modeling (hazard ratio (HR): 3.6; 95% confidence interval (CI): 1.9–6.8, p<0.001), after adjusting for demographic and cardiovascular risk factors. Furthermore, there was a significant and independent effect of high baPWV on stroke as shown among participants with adequate hypertension control (HR=2.3, 95%CI: 1.1–4.8, p=0.029).ConclusionAmong hypertensive patients, baPWV and hypertension control were demonstrated to independently and jointly affect the risk of first stroke. Participants with high baPWV and inadequate hypertension control had the highest risk of stroke compared to other groups. Our findings warrant additional investigation and raise a possibility that baPWV may serve as a simple noninvasive screening tool in clinical or community settings to identify hypertensive adults at high‐risk of stroke. It may also offer a novel therapeutic target to further reduce stroke risk beyond conventional management.