bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P155: Inadequate Blood Pressure Control in Hypertensive Patients Referred for Cardiac Stress Test
Authors: Mousa Tarek M , Akinseye Oluwaseun A , Kerwin Todd C
Published: July 2021

Introduction:
Hypertension (HTN) is a powerful risk factor for fatal and nonfatal cardiovascular events. Achieving adequate blood pressure (BP) control can reduce morbidity and mortality from cardiovascular diseases.The current study examined the degree of BP control and incidence of myocardial ischemia in hypertensive patients referred for cardiac stress test.

Methods:
We retrospectively analyzed 2,039 consecutive patients with the diagnosis of HTN referred to New York Hospital Medical Center of Queens/Weill Cornell Medical College nuclear cardiology laboratory for stress testing from January 2007 through July 2010. Patients were categorized into well-controlled (<140/90 mmHg), poorly-controlled (140-160/90-100 mmHg), and very poorly-controlled (>160/100 mmHg) groups according to their resting BP measured by an IntelliVue MP70 (Royal Philips Electronics, the Netherlands) non-invasive oscillometric BP monitor. The incidence of ischemia was defined as the presence of at least one reversible perfusion defect on stress/rest single photon emission computed tomography scan

Results:
Mean age [±SEM] = 68 ± 13 years, 885 (43.4%) were males. Prevalence of well-controlled HTN was 47.2%, poorly-controlled HTN, 29.5% and very poorly-controlled HTN, 23.3%. Evidence of ischemia was seen in 19.8% and 19.3% of the well-controlled and poorly-controlled group respectively. The very poorly-controlled group had the lowest incidence of ischemia (14.3%) (p < 0.05) compared to the other two groups.

Conclusions:
Symptoms mimicking ischemic heart disease in hypertensive patients may be partly explained by poorly controlled BP. Quality of care might be improved by optimally controlling BP in patient with angina symptoms prior to ordering diagnostic testing associated with radiation exposure and cost
Abstract P169: Antenatal Betamethasone Attenuates the Angiotensin-(1-7)/Nitric Oxide Axis in Adult Male but not Female Renal Proximal Tubule Cells
Authors: Su Yixin , Pulgar Victor M , Bi Jianli , Chappell Mark , Rose James C
Published: July 2021

Our studies have revealed a sex-specific effect of fetal programming on sodium (Na+) excretion in adult sheep whereby the males exhibit reduced Na+ excretion and an attenuated natriuretic response to Ang-(1-7) as compared to the females. We hypothesize that the renal proximal tubules are a key target for the early programming effects of glucocorticoids exposure to regulate Na+ handling in the adult males. Therefore, we isolated and cultured cortical proximal tubule cells (RPTC) from adult male and female sheep antenatally exposed to betamethasone (Beta) or vehicle. Na+ uptake and nitric oxide (NO) were assessed with Sodium Green and DAF fluorescence prior to and following a low dose of Ang-(1-7) (1x10-11 M) in isolated RPTC from sheep at ~1.5 years of age. Data are expressed as % of basal uptake or area under the curve (AUC) for Na+ or % of control for NO. Male Beta RPTC exhibit greater Na+ uptake than male vehicle cells (427±32%, n=13, vs. 315±28%, n=14, p<0.05; however, Beta had no effect on Na+ uptake in the female cells (242±18%, n=9, vs. 250±15%, n=10, p>0.05). Ang-(1-7) inhibited Na+ uptake in RPTC from vehicle male (255±40%) and from both vehicle (191±14%) and Beta (209±11%) females (Figure 1B), but failed to attenuate Na+ uptake in Beta male cells (Figure 1A). Beta exposure also abolished NO stimulation by Ang-(1-7) in male but not female RPTC (Figure 1C). We conclude that an Ang-(1-7)-NO-dependent pathway contributes to the sex-dependent consequences of programming on Na+ regulation in the proximal tubules of the kidney. Moreover, the RPTC retain both the sex and Beta-induced phenotype of the adult and may reflect an appropriate cell model of fetal programming.
Abstract P171: Mast Cell Inhibition Attenuates Cardiac Remodeling and Diastolic Dysfunction in Ovariectomized, Middle-aged Fischer344хBrown Norway Rats
Authors: Wang Hao , da Silva Jaqueline , Alencar Allan , Lin Marina S , Sun Xuming , Ahmad Sarfaraz , Ferrario Carlos M , Groban Leanne
Published: July 2021

The incidence of left ventricular hypertrophy (LVH) and diastolic dysfunction (LVDD) increases in postmenopausal women, but the mechanisms are not yet clear. This study determined the cardioprotective effects of chronic mast cell inhibition by the mast cell stabilizer, cromolyn sodium, in middle-aged (18-month-old), female Fischer344хBrown Norway (F344BN) rats after estrogen (E2) loss by ovariectomy (OVX). Eight weeks after OVX, systolic blood pressure increased in OVX vs. sham-operated rats (141±6 vs. 108±4 mmHg, P<0.05), and cromolyn treatment (30 mg/kg/day x 4 weeks, s.c. via osmotic minipump, n=7/group), initiated one month after OVX, attenuated this effect (115±4 mmHg). Myocardial relaxation (e') was reduced, LV filling pressures (E/e') were increased, and LV mass, wall thicknesses, and percent interstitial fibrosis were increased in OVX vs. sham rats. All of these cardiac adverse effects of E2 loss were mitigated by cromolyn treatment (Figure). Cardiac mast cell number was increased after OVX, irrespective of cromolyn. While no differences in plasma angiotensin (Ang) II levels were observed between OVX and sham rats (33.7±4.6 vs. 32.0±4.5 pg/mL), plasma levels of Ang II were reduced in cromolyn-treated OVX rats (21.3±3.0 pg/mL) (P<0.05 vs. sham and OVX-vehicle). Ang II content was significantly increased in hearts of OVX vs. sham rats, and cromolyn attenuated this effect. Moreover, cromolyn prevented the increase in cardiac Ang II type 1 receptor (AT1aR) mRNA expression in OVX rats. Our findings demonstrate that mast cell inhibition with cromolyn attenuates adverse LV remodeling and LVDD in OVX-F344BN rats possibly through a chymase/Ang II-mediated mechanism.
Abstract P172: Sex Differences in Obesity Associated Diastolic Dysfunction in Western Diet Fed Mice
Authors: Aroor Annayya , Habibi Javad , DeMarco Vincent G , Jia Guanghong , Garro Mona , Manrique Camila , Sowers James
Published: July 2021

Premenopausal women are protected against cardiovascular disease (CVD); however, this protection is lost in the setting of obesity, insulin resistance and type 2 diabetes. The mechanisms responsible for abrogation of the sex-related CVD protection are not clearly understood. We have recently developed a translational model in which female mice fed a diet high in fat and refined carbohydrates (western diet - WD) develop cardiac stiffness and diastolic dysfunction earlier than males consuming a WD. We hypothesized that these earlier adverse effects in females are mediated via increased mineralocorticoid (MR) activation/oxidative stress mediated activation (phosphorylation) of the serine kinase, S6K1 which promotes cardiac growth and fibrosis. Accordingly, four week old male and female C57BL6/J mice were fed a WD (containing high fat [46%], sucrose [17.5%], and high fructose corn syrup [17.5%]) or control diet (CD) for 8 and 16 weeks. Two-dimensional echocardiograms were used to evaluate diastolic function. Immunohistochemistry and western blotting were used to evaluate MR receptor expression and S6K1 phosphorylation. Diastolic dysfunction, indicated by prolonged isovolumic relaxation time (IVRT) and abnormal myocardial performance (increased myocardial performance index [MPI]) was present at 8 weeks in WD-fed female, but not male mice. Although male mice fed a WD exhibited diastolic dysfunction at 16 weeks, diastolic function as assessed by IVRT was more pronounced in female mice. The magnitude of cardiac fibrosis and oxidative stress was greater in females consuming a WD. Moreover, levels of plasma aldosterone, expression of MR (WD female 1.60 fold, WD male 1.26 fold), phosphorylation of S6K1 (WD females 2.92 fold, WD males 1.97 fold) and levels of mRNA for monocyte chemoattractant protein 1 (MCP-1, WD females 1.86 fold, WD males 1.16 fold) were higher in WD-fed female mice compared to WD-fed male mice. These results suggest enhanced cardiac MR mediated S6K1 activation and increased immune and inflammatory responses contribute to enhanced fibrosis and abrogation of cardiac protection in female mice fed a WD high in fat and fructose.
Abstract P187: The Salt Sensitivity of Human GRK4 Transgenic Mice Is Associated With Renal Oxidative Stress
Authors: Diao Zhenyu , Asico Laureano , Villar Van Anthony M , Zheng Xiaoxu , Cuevas Santiago , Armando Ines , Jose Pedro A , Wang Xiaoyan
Published: July 2021

Variants of hGRK4 gamma

are associated with human essential hypertension. hGRK4gamma

142V

transgenic mice are hypertensive without oxidative stress on normal salt intake. hGRK4 gamma

486V

transgenic mice have salt-sensitive hypertension. Because renal oxidative stress is increased in some rodent models of salt-sensitive hypertension, we quantified the renal expression of reactive oxygen species related proteins in
hGRK4
gamma

486V

transgenic mice and non-transgenic (NT) littermates, on normal (NS, 0
.
8%
)
and high (HS, 4%) NaCl diets
.
Systolic blood pressure (measured under anesthesia) was similar in
hGRK4
gamma

486V

(89.83±2.7, mm Hg, n=9) and NT (94.7±2.5) mice on NS diet and elevated in
hGRK4
gamma

486V

(114±6.1) but not in NT mice (94.1±2.8) on HS diet. The renal expressions of NOX1, 2, and 4 were similar in both strains on NS diet but NOX2 was decreased by HS in NT (28±7, % of NT on NS diet, n=5-6/group). On NS diet, CuZnSOD and ECSOD were similar in the two mouse strains while MnSOD (66±3%) was lower in
GRK4
gamma

486V

than NT mice. However, on HS diet, CuZnSOD (87±7%), MnSOD (70±3%), and ECSOD (55±7%) were decreased in
GRK4
gamma

486V

mice but not altered in NT mice. HO-2, not HO-1, was slightly greater in
GRK4
gamma

486V

than NT mice on NS diet (117±7%) but this difference was abolished by HS diet. Urinary 8-isoprostane was lower in
GRK4
gamma

486V

than NT mice (57±2.5 vs 70±0.1, ng/mg of Cr) on NS diet but increased to a greater extent in
GRK4
gamma

486V

than NT mice (197±18 vs128±6) on HS diet. Renal SOD activity and superoxide production were similar in both mouse strains on NS diet. HS diet decreased SOD activity (81.6±2.7%) and increased superoxide (138.9±6.6%) production in
GRK4
gamma

486V

mice but not in NT mice. The renal tubular immunofluorescence of ECSOD was reduced to a greater extent in hGRK4 gamma

486V

than NT by HS diet. HS diet also decreased the renal expression of NOS3, not NOS 1 and 2, in
GRK4
gamma

486V

mice (49±2%) but not in NT mice. These changes were not observed in the mice with hGRK4 gamma

wildtype

and hGRK4 gamma

142V

transgenic mice, suggesting that the salt-sensitive hypertension of
GRK4
gamma

486V

mice is related to renal oxidative stress.
Abstract P192: Both Mineralocorticoid Receptor and Angiotensin II type 1 Receptors in the Subfornical Organ Mediate Angiotensin II Induced Reactive Oxygen Species (ROS) in Brain Angiotensinergic Pathways
Authors: Wang Hong-Wei , White Roselyn , Huang Bing S , Chen Aidong , Ahmad Monir , Leenen Frans H
Published: July 2021

Activation of angiotensinergic pathways and central aldosterone (aldo)-MR-ENaC-endogenous ouabain (EO)-AT
1
R pathway play a critical role in Ang II associated hypertension. The SFO contains both MR and AT
1
R and can relay the signals of circulating Ang II to downstream nuclei such as the PVN, SON and RVLM. We evaluated the effect of knockdown of MR and AT
1
R specific in the SFO on reactive oxygen species (ROS) production in downstream nuclei. Wistar rats were intra SFO infused with AAV-MR- or AT
1a
R-siRNA and after 7 days received a sc infusion of Ang II at 500 ng/min/kg for 2 weeks. MR and AT
1
R expression were measured by real-time qPCR and western blotting. ROS was assessed by DHE staining.

Ang II increased AT
1
R mRNA expression in the SFO. Both MR- and AT
1
R-siRNA in the SFO prevented this increase. Ang II decreased MR mRNA but increased protein expression in the SFO. Both MR- and AT
1
R-siRNA further decreased MR mRNA expression. Ang II significantly increased ROS in the SFO, magno- and parvocellular parts of the PVN, SON and RVLM. Both MR- and AT
1
R-siRNA in the SFO prevented ROS increases in the PVN and RVLM. In contrast, MR- but not AT
1
R-siRNA in the SFO prevented the Ang II-induced ROS in the SON. Both MR- and AT
1
R-siRNA in the SFO prevented most of the Ang II-induced hypertension. These results suggest that aldo-MR signaling in the SFO is needed for the activation of Ang II-AT
1
R signaling from the SFO to the PVN and RVLM. Considering that only MR-siRNA in the SFO prevents circulating Ang II induced ROS in the SON, activation of aldo-MR signaling from the SFO to the SON may via EO enhance AT
1
R dependent activation of pre-sympathetic neurons in the PVN, and thereby to Ang II-hypertension.
Abstract P220: Congestive Heart Failure in the Rat Induces Subtle Renal Damage via Neurogenic Pathways
Authors: Ditting Tilmann , Linz Peter , Hindermann Martin , Rodionova Kristina , Heinlein Sonja , Ott Christian , Schmieder Roland E , Hilgers Karl F , Amann Kerstin , Veelken Roland
Published: July 2021

Background:
Cardiomyopathy in experimental renal insufficiency is putatively influenced by neurogenic pathways of renal origin. We wondered if cardiac neurogenic effects in congestive heart failure could likewise harm the kidney. We hypothesized that increased renal sympathetic nerve activity (RSNA) in rats with congestive heart failure after myocardial infarction (CHF) induces renal structural damage.

Methods:
21 day after induction of CHF renal morphology was evaluated by immunohistology (interstitial and glomerular mononuclear cell infiltration (ED1), cell proliferation (PCNA), collagen I,III,IV,V,VI, laminin und fibronectin). RSNA was assessed by volume challenge (VE) to decrease RSNA. CHF and control rats were investigated with and without renal denervation (DNX). Blood pressure (BP), heart rate (HR) and RSNA were recorded. Nodose ganglion neurons (NGN) with vagal cardiac afferents were cultured for 1 day. Whole cell recordings were obtained and current-voltage relationships established. Cells were characterized by osmomechanical stress with a mannitol solution.

Results:
In CHF rats with intact renal nerves (nonDNX) formation of collagen I occurred, that was reduced after DNX (12.2+0.7 %area vs. 9.1+1.1 %area*, n=6, * p<0.05). VE-induced RSNA decreases were impaired in CHF vs controls suggesting increased RSNA (-α 34+8% vs. -α[[Unable to Display Character: ]]54+6% *, n=6, * p<0.05). NGN from CHF exhibited altered conductance in response to mechanical stress as compared to controls (change in holding current at -80 mV: control_normoosmotic: -144±30 pA; control_hypoos.: -282±34 pA vs CHF_normosmotic: - 230±55 pA; CHF_hypos.: -540±100* pA; *p<0.05 CHF vs. control).

Conclusion:
CHF induced subtle renal structural damage due to increased renal sympathetic tone which was likely due to altered NGN mechanosensitivity. Afferent nerve units from cardiovascular organs obviously form a complex sympathomodulatory network.
Abstract P222: Venous Dilation Contributes to 5-HT-induced Hypotension
Authors: Seitz Bridget M , Krieger-Burke Teresa , Watts Stephanie W
Published: July 2021

Serotonin (5-hydroxytryptamine, 5-HT) infusion in a normal conscious rat decreases mean arterial pressure (MAP), in part by reduction in total peripheral resistance. Microsphere experiments have shown 5-HT increases blood flow within the splanchnic vascular bed, with the greatest being in the intestine and spleen. Interestingly, 5-HT does not cause a direct relaxation of resistant (small or large) mesenteric arteries. The present study addresses the possibility of the venous circulation contributing to the 5-HT induced fall in blood pressure. Our working hypothesis is venous dilation, specifically dilation of veins measurable within the splanchnic vascular bed, contributes to 5-HT-induced hypotension. Using an ultrasound imaging system (Vevo 2100 imaging system; 21 MHz probe,Visual Sonics Inc.), telemetry-implanted, anesthetized male Sprague Dawley rats underwent cross-sectional imaging which was controlled for respiration and cardiac cycles. The following vessels were imaged: abdominal aorta (AA); portal vein (PV); abdominal inferior vena cava (IVC); and superior mesenteric vein (SMV). Following the collection of baseline MAP and vessel diameter measurements, Alzet osmotic mini-pumps containing vehicle (saline; n=9) or 5-HT (25 ug/kg/min; n=9) were implanted for 1 week. After, 24 hours of infusion, 5-HT increased the vein diameter (SMV 17.48±2%; PV 17.67±2%; IVC 46.87±8%) and maintained the AA diameter ( AA 0.93±1%) from baseline while reducing MAP (vehicle 101.93±3; 5-HT 84.68±2 mm Hg; p<0.05).One-week post removal of all osmotic mini-pumps, there was no difference in the MAP or diameter of all noted vessels between the two treatment groups. To correlate with in vivo findings, the PV and IVC, when isolated in a tissue bath for measurement of isometric force and contracted with endothelin 1, relaxed in a concentration dependent fashion to 5-HT and 5-carboxamidotryptamine (5-HT 1/7 receptor agonist;1 nM-10 uM). Collectively, these findings highlight the contribution of splanchnic venous dilation in 5-HT-induced hypotension and propose a possible mechanism for 5-HT reduction in blood pressure.
Abstract P229: Is a Multidisciplinary Approach to Hypertension Better Than Traditional Care in Female Hypertensive Patients?
Authors: Weber Jessica A , Shah Shital C , Turley Sara , Braun Lynne T , Kent Erica R , Volgman Annabelle S
Published: July 2021

Background:
Rush Heart Center for Women (RHCW) opened in October 2003 to provide a multidisciplinary approach (MDA) for female patients (pts). RHCW provided personalized care plans to address women’s heart health, with an emphasis on female-specific risk factors and symptoms. MDA including cardiologists, dietitians and nurse practitioners to treat female pts was compared to a similar cohort using standard practice in terms of HTN treatment.

Methods:
A retrospective study identified pts with HTN treated at RHCW using a MDA and compared to a cohort of pts treated by cardiologists and received a standard level of care consistent with guidelines and recommendations (standard practice). Pts seen between 2008 and 2014 with at least 2 visits between 9 and 15 months apart were studied. Demographic moderating variables were race, age, and insurance. Moderating clinical variables were baseline systolic BP, body mass index, diabetes, smoking status, history of coronary disease, stroke, and prior treatment of HTN. Bivariate and multivariate analyses were conducted to determine the effect of treatment type, with pts’ follow-up BP.

Results:
A total of 1486 pts were evaluated. Pts seen treated by MDA was younger by <3 years. Table shows descriptive statistics and bivariate analysis. Multivariate analysis revealed that pts treated with MDA had a significantly lower BP in their follow-up systolic BP, 3.8 mmHg less, compared to pts treated with standard practice. (p<0.002).

Conclusion:
The multidisciplinary approach had better BP control in female hypertensive pts. Prospective studies comparing MDA to standard practice may help to assess improved quality of life, compliance and outcomes.
Abstract P232: Dipeptidyl Peptidase-4 (DPP-4) Inhibitor, Linagliptin, Prevents Aortic Stiffening and Vascular and Cardiac Diastolic Dysfunction Caused by Western Diet Feeding in Female Mice
Authors: DeMarco Vincent G , Aroor Annayya , Jia Guanghong , Habibi Javad , Garro Mona , Sun Zhe , Meininger Gerald , Klein Thomas , Whaley-Connell Adam , Sowers James
Published: July 2021

Aortic stiffness, endothelial dysfunction and diastolic dysfunction (DD) are cardiovascular (CV) abnormalities seen in obesity associated with consumption of high fat/fructose western diet (WD). Moreover, CV dysfunction is increasingly prevalent in obese women. Herein, we examined whether the DPP-4 inhibitor, linagliptin (LINA), improves these outcomes in WD fed female C57BL/6 mice. Four week old mice were fed control diet (CD) or WD with or without LINA for 16 weeks, after which pulse wave velocity (aortic stiffness) (PWV), echocardiography (diastolic function), atomic force microscopy (endothelial stiffness) and wire myography (aortic vascular reactivity) were performed. Compared to CD mice, WD mice exhibited 21% and 353% higher PWV and endothelial stiffness, respectively. WD induced DD, indicated by impaired septal wall motion (E/Vp ratio), prolonged isovolumic relaxation time (IVRT) and impaired myocardial performance index (>MPI). These vascular and cardiac abnormalities were prevented by LINA. LINA also prevented WD-induced impairments in acetylcholine-, sodium nitroprusside-, and insulin-mediated aortic vascular relaxation. These results show that LINA exerts CV protection in a translational model of obesity.
Abstract P244: Role of the Na-H Exchanger Regulatory Factor 1 (NHERF1) in Hypertension of Aging Animals
Authors: Pushpakumar Sathnur , Ketchem Corey J , Barati Michelle T , Sen Utpal , Jose Pedro J , Weinman Edward J , Lederer Eleanor D , Khundmiri Syed J
Published: July 2021

Aging animals develop hypertension when challenged with high salt diet due, in part, to desensitization of dopamine receptors (DR) in renal proximal tubules (RPT). We have demonstrated that NHERF1 associates with DR1 and Na-K ATPase (NKA) and is important for regulation of NKA in RPT. Preliminary data showed loss of NHERF1 expression in 22m old F344 rats. We hypothesized that loss of NHERF1 results in increased blood pressure (BP) and lack of natriuretic response to dopamine (DA) in aging animals. To address this hypothesis, Fischer Brown Norway (FBN) rats (1m, 4m, 12m, and 24m old) were fed diet containing 1% or 8% NaCl for one week and, BP was measured in anesthetized animals using an indwelling left femoral artery catheter. 8% NaCl did not increase BP in 1m or 4 month old rats. By contrast, 8% NaCl diet increased BP in 12m (84.3±3.5 vs 90.8±2.36) and 24m (73.5±7.58 vs 104±1.6) old animals. To determine if lack of NHERF1 is responsible for the increase in BP, we measured BP in 12 m old WT and NHERF1 KO mice. By contrast to WT mice, 8% NaCl diet did not increase BP in NHERF1 KO mice (84±4.9 vs 96.5±3.56 (WT) and 78.2±3.89 vs 81.8±9.2 (NHERF1 KO mice)). To confirm that NHERF1 is required for DA-mediated inhibition of NKA, NKA activity in primary proximal tubule cells (PTC) from young and old mice in culture was measured in the presence or absence of DA. DA decreased NKA activity in PTC from young animals (67.2±3.8 vs 32.7±5.3) but not in PTC from old animals. Transfection of NHERF1 restored NKA regulation by DA in PTC from old rats (58.4±4.2 vs 64.4±4.3 (in untransfected cells) 54.2±3.8 vs 31.1±3.4 (in NHERF1 transfected cells)). We conclude that NHERF1 regulates DA-mediated proximal tubule sodium handling; however, other factors modulate BP response to dietary sodium intake in young and old animals. The contribution of NHERF1 and dopamine signaling to sodium homeostasis requires further study.
Abstract P251: 3-hydroxy-3-methylglutaryl-coenzyme A reductase Inhibition Reduces Intimal Neovascularization and Plaque Growth in Apolipoprotein E-Deficient Mice
Authors: Cheng Xian Wu , Hu Lina , Inoue Aiko , Wu Hongxian , Jiang Haiying , Li Xiang , Okumura Kenji , Kuzuya Masafumi
Published: July 2021

Objective:
The interactions between the renin-angiotensin system and neovascularization in atherosclerotic plaque development are unclear. We investigated the effects of 3-hydroxy-3-methylglutaryl-coenzyme a reductase inhibition with pitavastatin in the pathogenesis of atherosclerosis in ApoE-/- mice with special focus on plaque neovascularization.

Methods and Results:
Ten-week-old male ApoE-/- mice fed a high-fat diet were randomly assigned into two groups and administered vehicle (0.5% carboxymethyl cellulose) or pitavastatin (PiS, 1 mg/kg daily) for 12 weeks. Quantification of plaque areas at the aortic roots and in the thoracic and abdominal aortas revealed that, in all three regions, AT1R antagonism reduced the intimal neovessel density and the mRNA levels of toll-like receptor (TLR) 2 and TLR4. PiS increased the contents of collagen and elastin, lessened the macrophage component as well as the level of monocyte chemoattractant protein-1 and osteopontin protein in aortic roots, and reduced the mRNA and activity levels of matrix metalloproteinase (MMP)-2 and MMP-9 in aortic roots and thoracic aortas. Neointimal vessel density, the extent of atherosclerotic lesions, and the levels of TLR2 and TLR4 mRNA, were lower in ApoE-/-MMP-2-/- mice than in controls. The amounts of TNF-α and IL-1β protein as well as the levels of MMP2, MMP-9, TLR2, and TLR4 mRNA were increased by exposure to ox-LDL and lipopolysaccharide. These effects were diminished by PiS and small interfering RNAs targeting TLR2 or TLR4 in cultured macrophages.

Conclusions:
3-hydroxy-3-methylglutaryl-coenzyme a reductase inhibition appears to inhibit intimal neovascularization in ApoE-/- mice, partly by reducing TLR2/TLR4-mediated inflammation and MMP activation, thus decreasing atherogenic plaque growth and instability.
Abstract P260: Role of Primary Endothelial Cilia in Hypertension
Authors: Saternos Hannah , Hossain Md Zubayer , AbouAlaiwi Wissam
Published: July 2021

Primary cilia are mechanosensory organelles that are projected into the lumen of blood vessels. It has been demonstrated that vascular endothelia require primary cilia to sense and transmit external mechanical stimuli into internal biochemical reactions. One of these reactions includes the biosynthesis and release of nitric oxide, which is one of the most potent endogenous vasodilators. This idea has
only
been investigated in cultured endothelial cells
in vitro
. Based on this finding, however, a very bold hypothesis is formed to test that abnormal cilia function results in vascular hypertension. Our laboratory has recently generated and obtained several conditional mouse models to specifically study the function and structure of primary cilia in vascular endothelia. These models include
1)
mice without cilia function (
Pkd1
or
Pkd2
);
2)
mice without cilia structure (
Tg737
or
Kif3a
). Our data indicate that mice with abnormal cilia function (
Pkd1
) or structure (
Tg737
) show significantly higher systolic (150±19 for

Pdgfbcre:Pkd1
flox/flox

and 147±10 for

Tie2Cre:Tg737
flox/flox

vs. 128±9 for wild-type) and diastolic (120±21 for

Pdgfbcre:Pkd1
flox/flox

and 120±11 for

Tie2Cre:Tg737
flox/flox

vs. 102±7 for wild-type) blood pressure than the corresponding wild-type mice. Because there is a positive and continuous correlation between blood pressure and cardiovascular diseases, satellite hypotheses are developed to look at the pathophysiological roles of endothelial cilia in cardiac functions and focal vascular diseases
in vivo
. Our data clearly point towards deteriorating phenotypes in the cardiac muscle, including cardiac fibrosis due to an increased cardiac workload. As a result, a heart-to-body weight ratio was significantly increased by 17 weeks old (0.008

PdgfbCre;Pkd1
f/f

vs. 0.006

Pkd1
f/f

).The present study will very likely provide new insights for hypertension and offer advanced scientific understanding of vascular endothelial cilia in other cardiovascular diseases.
Abstract P601: Matrix Metalloproteinase 2 Plays An Important Role In Angiotensin Ii-induced Vascular Injury Mediated In Part Through Epidermal Growth Factor Receptor Activation In Vascular Smooth Muscle Cells
Authors: Barhoumi Tlili , Mian Muhammad Oneeb Rehman , Fraulob-Aquino Julio C. , Rehman Asia , Idris-Khodja Nourredine , Paradis Pierre , Schiffrin Ernesto L.
Published: July 2021

Background:
Matrix metalloproteinase 2 (MMP2) is involved in cardiovascular disease. Whether MMP2 plays a role in hypertension and vascular damage is unknown. We hypothesized that Mmp2 knockout will prevent angiotensin (Ang) II-induced blood pressure (BP) rise and vascular injury.

Methods:
Ten to 12-week-old male Mmp2 knockout (Mmp2-/-) and wild-type (WT) mice were infused with Ang II (1000 ng/kg/min, SC) for 14 days. Systolic BP was measured by telemetry, mesenteric arteries (MA) endothelial function and vascular remodeling by pressurized myography. In aortic wall or perivascular fat (PVAT), reactive oxygen species (ROS) generation was determined using dihydroethidium staining, and vascular cell adhesion protein 1 (VCAM-1), monocyte chemotactic protein-1 (MCP-1) expression and monocyte/macrophage infiltration by immunofluorescence. Spleen T cells and monocyte profile were assessed by flow cytometry. Vascular smooth muscle cells (VSMCs) were isolated from MA of WT and Mmp2 knockout mice, stimulated 5 min with 100 nM Ang II and epidermal growth factor receptor (EGFR) phosphorylation measured by Western-Blotting.

Results:
Ang II increased Systolic BP (172±7 vs 122±3, P<0.01), decreased MA vasodilatory responses to acetylcholine (33±5% vs 83±3%, P<0.01) and increases MA media-to-lumen ratio (5±0% vs 3±0%, P<0.01), media cross-sectional area (7224±467 vs 5345±336 μm2, P<0.05), and stiffness (P<0.01), as shown by a leftward shift of the stress/strain relationship, in WT. Furthermore, Ang II enhanced aortic ROS generation (73±11 vs 6±1 RFU/μm2, P<0.01), aortic VCAM-1 (17±3 vs 5±3 RFU/μm2, P<0.01) and MCP-1 expression (71±14 vs 11±3 RFU/μm2, P<0.01) and PVAT monocyte/macrophage infiltration (32±5 vs 4±0 RFU/μm2, P<0.05), and spleen activated CD4+CD69+ and CD8+CD69+ T cells and pro-inflammatory Ly-6Chi monocytes (17±2 vs 10±1%, 11±1 vs 5±1% and 53±6 vs 25±2%, respectively, P<0.05) in WT. Ang II increased EGFR phosphorylation in VSMCs in vitro (1.91±0.19 vs 1.0±0.0%, P<0.05). Mmp2 knockout prevented or reduced all of the above except BP elevation (P<0.05).

Conclusion:
MMP2 plays an important role in Ang II-induced vascular injury, which could be mediated at least in part through EFGR activation in VSMCs.
Abstract P603: Age and Angiotensin-(1-12) Expression in Human Atrial Tissue of Patients with Left Heart Disease
Authors: VonCannon Jessica , Varagic Jasmina , Nagata Sayaka , Ahmad Sarfaraz , Locke Adair , Wang Hao , Groban Leanne , Kon Neil , Ferrario Carlos M
Published: July 2021

In the human heart formation of angiotensin (Ang) II results from the hydrolysis of an alternate angiotensin substrate, -Ang-(1-12)-, by chymase rather than angiotensin converting enzyme. In our recent study a higher Ang-(1-12) expression and upregulation of chymase mRNA and enzymatic activity was found in left (LAA) versus right atrial appendages (RAA) of subjects with left heart disease. Since aging is associated with prominent changes in cardiac structure and function, we assessed the relationships among age, Ang-(1-12), and chymase gene expression and activity in both LAA and RAA in 44 patients undergoing cardiac surgery for the correction of valvular heart disease, resistant atrial fibrillation or ischemic heart disease. Immunohistochemistry for Ang-(1-12) detection was performed using an affinity purified polyclonal antibody directed to the COOH terminus of the full length of the sequence of human Ang-(1-12). Quantitative real-time polymerase chain reaction was used to detect chymase mRNA levels whereas chymase activity was assessed by HPLC. We report that Ang-(1-12) immunostaining in atrial appendages (r=0.30; p<0.05), but not chymase mRNA expression or activity, correlated directly with patients age. While a tendency for higher Ang-(1-12) expression in LAA (Intensity: 5.88 ± 0.91 units; n=11) when compared to RAA (Intensity: 3.948 ± 0.55 units; n=15) was noted in patients younger than 65 yrs, this difference was more prominent and statistically significant in patients older than 65 yrs of age (LAA Intensity: (n=12): 7.39 ± 1.06 units versus RAA Intensity (n=13): 4.74 ±0.54 units; p < 0.05). The results of the present study suggest an age-related increase in Ang-(1-12) expression in human atrial tissue that may be more prominent in the LAA of patients with left heart disease. We suggest that higher availability of Ang-(1-12) for direct Ang II formation may be an underlying mechanism responsible, at least in part, for age- and disease-related atrial and ventricular remodeling and dysfunction.
Abstract P605: Liver-specific Antisense Inhibition Of Angiotensinogen Reduced BP In A Hypertensive Rat Model Resistant To Standard RAS Inhibitors
Authors: Mullick Adam E , Yeh Steve T , Graham Mark J , Crooke Rosanne M
Published: July 2021

Uncontrolled hypertension is an important contributor to cardiovascular disease. Despite the armamentarium of antihypertensive therapeutics, there remains a need for a novel agent effective in individuals with treatment-resistant hypertension who cannot reach their target BP. Pharmacological agents targeting the renin-angiotensin-aldosterone system (RAAS) are widely used but may not optimally inhibit RAAS. Experiments were conducted to characterize a series of angiotensinogen (Agt) oligonucleotides (ASOs) and compare their efficacy in a model of malignant hypertension. Agt ASOs which targeted all systemic sites of Agt (e.g. liver, fat and kidney) versus LICA-conjugated Agt ASOs (Agt-LICA ASOs) that preferentially target the liver were evaluated in normotensive and hypertensive rats. Liver, adipose and kidney Agt RT-PCR were performed and revealed Agt ASO treatment reduced liver, adipose and kidney Agt expression by 93%, 96% and 82%, respectively. Agt-LICA ASO treatment resulted in 93%, 10% and 11% reductions of Agt mRNA in liver, adipose and kidney, respectively, thus demonstrating specific liver targeting with LICA. Plasma Agt, measured by ELISA, revealed no significant differences between the Agt and Agt-LICA ASO treatments. For example, after a single administration of the ASOs, plasma Agt was reduced 15-18%, 41-48%, 84-89% and 92% at 12, 24, 48 and 72 hours, respectively, after ASO administrations. Radiotelemetry was used to compare the BP lowering efficacy in spontaneously hypertensive rats (SHRs) fed an 8% NaCl diet, a model of malignant hypertension resistant to standard RAAS inhibitors for BP control. Treatments of a control ASO or captopril or captopril plus losartan had a modest effect on BP whereas treatments with either the Agt or Agt-LICA ASOs resulted in similar MAP reductions of 30 - 40 mm Hg. These data demonstrate that the effects of global vs. liver-specific Agt inhibition result in similar reductions of plasma Agt or blood pressure. Additionally, ASO treatments produced superior BP efficacy relative to the standard of care in a model resistant to RAAS blockage. Such improvements could be desirable in individuals not at their BP goal with existing RAS therapeutics.
Abstract P611: Vascular-mediated Preterm Birth is Associated with Cardiovascular Risk after Pregnancy
Authors: Catov Janet M , Reis Steven E , Muldoon Matthew F , Ness Roberta B , Nguyen Lananh , Parks W. T
Published: July 2021

Objective:
To investigate whether preterm birth and placental evidence of malperfusion is associated with subclinical atherosclerosis and a higher cardiovascular risk factor burden 4 to 12 years after pregnancy.

Methods:
A cohort of women with preterm (n=119) and term births (n=242), mean age 38 years, was examined on average eight years after pregnancy for carotid artery intima-media thickness (IMT), fasting lipids, blood pressure and inflammatory markers (C-reactive protein [hsCRP] and Interleukin-6 [IL-6]). Pregnancy characteristics included placental pathology evidence of malperfusion (vasculopathy, infarct, advanced villous maturation, perivillous fibrin, fibrin deposition), infection (chorioamnionitis, funisitis, deciduitus), villitis (chronic inflammation), fetal thrombosis or chorangiosis. Vascular-mediated preterm births were those with malperfusion lesions, and by design, those with preeclampsia were excluded.

Results:
Women with malperfusion lesions had a higher mean carotid IMT (+0.055 cm), total cholesterol (+17.49 mg/dl), LDL-C (+11.44), triglycerides (+17%), apolipoprotein-B (+8.95) and systolic and diastolic blood pressure (+4.58/+2.62 mmHg) compared to women with term births, independent of age, race, smoking and adiposity assessed before and after pregnancy (all p<0.05). Women with preterm birth and evidence of malperfusion accompanied by other lesions related to infection or chronic inflammation had the most atherogenic profile after pregnancy, and carotid IMT differences were independent of traditional risk factors (+0.04 cm; p=0.027).

Conclusions:
Vascular-mediated preterm birth is associated with maternal subclinical atherosclerosis and a higher cardiovascular risk factor burden in the decade after pregnancy compared to term birth. The placenta may offer unique insight into how pregnancy complications can portend the emergence of maternal cardiovascular disease.
Abstract P613: The Effect of Metabolic Factors on Hypoxia-Induced sFlt-1 Secretion in Rat Placental Villi
Authors: Palei Ana C , Spradley Frank T , Granger Joey P
Published: July 2021

Although the etiology of preeclampsia (PE) remains unclear, evidence indicates that impaired trophoblast invasion followed by placental ischemia/hypoxia promotes the release of placental anti-angiogenic factors, such as soluble fms-like tyrosine kinase-1 (sFlt-1), into the maternal circulation. sFlt-1 blocks the pro-angiogenic actions of vascular endothelial growth factor to elicit maternal endothelial dysfunction and ultimately hypertension. Obesity is a major risk factor for PE. In addition, increased circulating metabolic factors, such as leptin and insulin have been associated with PE. However, the mechanisms whereby obesity and its related metabolic factors increase the risk for the development of PE are unknown. The aim of this study was to evaluate whether chronic leptin or insulin exposure exacerbate hypoxia-induced sFlt-1 secretion from rat placental villi. In order to address this question, placental villous explants were isolated from placentas of normal pregnant rats (n=4, 3 placentas per rat) and pregnant rats treated with either leptin (0.5 mg/kg/min i.p.; n=3, 3 placentas per rat) or insulin (1.5 mU/kg/min s.c.) supplemented with 20% glucose in drinking water (n=3, 3 placentas per rat) from gestational day 14 to 19. Placental explants were then incubated for 48 h at 37 °C under normoxia (6% O2) or hypoxia (1% O2) and sFlt-1 secretion in cultured media was measured by ELISA. While hypoxia significantly enhanced sFlt-1 release of explants from normal pregnant rats compared with normoxia (3224±224 vs 4251±236 pg/mg; P<0.05), explants from chronic hyperleptinemic (3197±178 vs. 3762±317 pg/mg) or euglycemic hyperinsulinemic (4066±186 vs. 4251±213 pg/mg) pregnant rats secreted similar sFlt-1 levels under normoxic and hypoxic conditions, respectively. Additionally, chronic leptin or insulin treatments did not exacerbate the effect of hypoxia on sFlt-1 release. In conclusion, our in vitro studies with placental villi from chronic hyperleptinemic or euglycemic hyperinsulinemic pregnant rats showed no exacerbation of hypoxia-induced sFlt-1 secretion.
Abstract P615: Hypertension-Dependent Reprogramming of the Endothelial Transcriptome
Authors: Nelson Jonathan W , Barnes Anthony P , Kaul Sanjiv , Ellison David H
Published: July 2021

Hypertension has drastic consequences for the cardiovascular system and especially for endothelial cell health. Clinically this is observed through the repeated finding that hypertension is associated with endothelial dysfunction. Despite this strong evidence that hypertension leads to impairment of endothelial function, the molecular events that give rise to endothelial dysfunction are poorly characterized. To better understand how hypertension affects the health and function of endothelial cells at the molecular level, we utilized a mouse model of spontaneous hypertension (BPH/2J) with the overarching hypothesis that hypertension causes the transcriptional reprogramming of endothelial cells leading to endothelial dysfunction. First, we non-invasively quantified endothelial function from hypertensive and normotensive (BPN/3J) mice by measuring flow-mediated vasodilation with high-frequency ultrasound. We found that hypertensive mice failed to dilate their femoral artery while normotensive control mice dilated their femoral artery 17 ± 8% (Mean ± SEM). Next, we acutely isolated cardiac endothelial cells by magnetic-assisted cell sorting for endothelial marker CD31. Through this technique, we enriched our sample for endothelial cells (as defined by expression of two endothelial-specific cell-surface markers CD31 and CD102) to 92 ± 1% compared to 9 ± 1% in pre-magnetically sorted cells. Finally, we isolated RNA and transcriptionally profiled the endothelial cells from normotensive and hypertensive mice (n=3 mice per group) with an average depth of 30 million reads per sample with RNAseq. We found that over 4000 genes were differentially expressed between groups (FDR cutoff of .01). Using Ingenuity Pathways Analysis (Qiagen), we identified multiple pathways that were dysregulated in endothelial cells exposed to hypertension including those related to immune function, cell morphology and cell-to-cell communication. This work represents one of the first studies to utilize cell-specific RNAseq to quantify how endothelial cells are transcriptionally reprogrammed in a pathological state. It is our hope that this work will lead to targeted therapeutics to improve endothelial function in patients with hypertension.
Abstract P622: Regulation of Neuroprotective Myeloid Cell Leukemia 1 by Rapamycin and AT2R Agonists in Dopaminergic Neuronal Cell-line
Authors: Bajwa Jamal , Pulakat Lakshmi
Published: July 2021

Myeloid Cell Leukemia I (MCL-1) is a critical protein for neuronal cell survival. MCL-1 is one of the anti-apoptotic proteins in the Bcl2 family. In neurons, MCL-1 regulates the rate of programmed cell death during development and after neuronal damage. It is now well established that without sufficient MCL-1 dopaminergic neuronal cells succumb to cell death under conditions of oxidative stress that result in neurodegenerative diseases such as Parkinsonism. Therefore, identifying drugs that can up-regulate the expression of MCL-1 in neuronal cells is critical for enhancing neuronal resistance to oxidative stress and improving neuronal survival. Aim of this study was to evaluate the effects of treatments with Rapamycin and a novel AT2R peptide agonist NP-6A4 on the MCL-1 expression in SH-SY5Y neuronal cell line. SH-SY5Y cells are a human-derived in vitro model of neuronal function and differentiation, expressing both adrenergic and dopaminergic markers. This cell line is a highly translational model for Parkinson's disease. Cells were maintained in a 1:1 mixture of DMEM and Ham's F-12 with 10% FBS. Cells were subjected to serum starvation and were treated with Rap (10nM), NP-6A4 (300nM) or their combination for 6 hours. MCL-1 protein expression was assessed by immunofluorescence using anti-MCL-1 antibody and a fluorophore-conjugated secondary antibody. Cells were imaged using a confocal microscope and fluorescence was quantified using Leica LAS AF software. It was observed that Rap treatment significantly suppressed MCL-1 expression in SH-SY5Y cells (~40% suppression, p<0.001), whereas Rap+NP-6A4 treatment reversed the Rap-mediated suppression of MCL-1 (p<0.0002). This data indicates that Rapamycin suppresses MCL-1 in dopaminergic neuronal cells and AT2R agonist, NP-6A4 is capable of reversing this effect.
Abstract P634: The Influence Of Virtual Learning Environment On Therapy Adherence And On The White Coat Effect
Authors: Fistarol Isabela R , Wen Chao L , Bortolotto Luiz , Vieira Margarida , Tsunemi Miriam , Giorgi Dante , Lopes Heno F , Consolim-Colombo Fernanda , KowalsKi Ivonete S , Guerra Grazia M
Published: July 2021

Systemic arterial hypertension is a risk factor for cardiovascular diseases and has become a common public health problem. Health education associated with educational technology may be used to encourage patients’ adherence to treatment and enable them to adequately understand how harmful hypertension can be to health, thereby promoting their quality of life. OBJECTIVE: To evaluate the influence of a strategy in an individual orientation program using educational technology associated with virtual learning environment (VLE) of hypertension care on the reduction in the white coat effect and the improvement in blood pressure control to be promoted by a nurse in a hypertension unit in a government state hospital in São Paulo. METHODS AND MATERIALS:This was a randomized clinical education study conducted with two groups, the VLE group (study group, 10 patients) and the control group (16 patients). Both groups were interviewed 6 times by nurses during the 120-day follow-up at 20-day intervals. At baseline (randomization) and at the end of the study, the patients took Spielberg’s State-Trait Anxiety Inventory (STAI), the Morisky test, and the WHOQOL, a quality of life instrument, and had their blood pressure taken (ambulatory blood pressure monitoring [ABPM]). Both groups had their blood pressure, weight, and abdominal circumference measured. Only the study group had remote access to the VLE. This consisted of 6 specific educational modules, each released according to the encounter number. RESULTS: At baseline, there were no statistical differences between the two groups with respect to the sociodemographic and hemodynamic variables. At the end of the study, there was a significant statistical difference between the groups on the Morisky test (p=0.001) and on the WHOQOL with respect to domain 3 social (p=0.001). There was no statistical difference with respect to the white coat effect between the groups. Nor was there any statistical difference between the groups with respect to the association of the anxiety degree measured by STAI and the white coat effect.CONCLUSION: In light of the results, our strategy improved the quality of life in the social domain and changed the adherence behavior of the study group in relation to the forgetfulness of medication schedules.
Abstract P641: Angiotensin Receptor Blocker Improves Pressure Natriuresis in Select African-Americans
Authors: Harshfield Gregory A , Hanevold Coral , Stewart Deborah , Dong Yanbin , Mathur Sunil
Published: July 2021

Objective:
Hypertension in ~30-60% of African-Americans (AAs) is resistant to single or combination therapy, indicating the necessity for novel screening strategies in this population. Our research supports a strategy that identifies a subset of AAs (approximately 1 in 3) who increase their retained sodium load during mental stress (MS) with an accompanying volume-mediated increase in blood pressure (BP) that remains elevated until the volume expansion diminishes. We hypothesize is due to activation of the renin-angiotensin system (RAS).

Design and Method:
We conducted a double blind placebo controlled crossover trial comparing the effects of irbesartan to placebo on sodium handling and consequent BP during MS. Subjects included 213 AA adults, of which 168 (78%) completed the study. After 7 days of vehicle (placebo or irbesartan) subjects underwent a 3 hr protocol comprised of 1 hr rest, 1 hr competitive video games, and 1 hr recovery. Blood and urine were obtained after each hour and BP every 15 mins. A 1 week washout ensued before subjects crossed over to repeat with the alternate vehicle. Subjects were divided into those who exhibited expected increases in urinary sodium excretion (U
Na
V) (Excreters) or reduced U
Na
V (Retainers) during MS as defined by previous data. Statistical analysis was performed using crossover methods with a pooled analysis that assumes equal variances and the Satterthwaite test of treatment difference (Stest) that assumes unequal variances.

Results:
Both statistical tests revealed that U
Na
V increased significantly for retainers (n=55) on irbesartan (p<0.002 and p<0.015, mean ± SD (pooled =7.15 ± 6.78, Stest =7.15)) normalizing the sodium response to MS. Irbesartan also enhanced U
Na
V during MS in excreters (n=113) with significant differences (p<0.001 and p<0.001, mean ± SD (pooled = 4.18 ± 9.03, Stest =4.18)). Overall SBP response was significantly reduced with treatment (p< 0.001 and p< 0.001, mean ± SD (pooled = -5.08 ± 3.19, Stest = -5.08)).

Conclusion:
Agents that block the RAS are not usually considered effective in AAs. However, our results suggest identification and treatment of AAs who increase their retained sodium load during MS with these agents will reduce the percentage of AAs with hypertension resistant to standard therapy.