bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 335: Prostaglandin I
2
Receptor Blockade Contracts Rat Renal Arteries and Facilitates Endothelium-Dependent Contractions.
Authors: Baretella Oliver , Xu Aimin , Vanhoutte Paul M
Published: March 2022

Endothelium-dependent contractions are augmented in human hypertension and in isolated preparations, including renal arteries, from hypertensive animals. Renal vasomotor tone is essential in the regulation of arterial blood pressure and renal arteries are highly sensitive to vasoconstrictor prostanoids including prostacyclin, identified as endothelium-derived contracting factor (EDCF) activating thromboxane prostanoid (TP) receptors. However, the mechanisms underlying increased EDCF responses in hypertension are not completely resolved. Isometric tension was measured, in wire myographs, in renal arterial rings from spontaneously hypertensive (SHR) and age (38-52 weeks)-matched Wistar Kyoto (WKY) rats. Responses were compared under control conditions and after incubation with indomethacin (10
-5
mol/L, cyclooxygenase inhibitor), ketanserin (10
-6
mol/L, 5-HT
2
receptor antagonist), CAY10441 (10
-6
mol/L, prostaglandin I
2
[IP] receptor antagonist), and S18886 (10
-7
mol/L, TP receptor antagonist). All experiments were performed in the presence of L-NAME (3 x 10
-4
mol/L) to exclude effects of endothelium-derived NO. Surprisingly, CAY10441 (10
-10
to 10
-6
mol/L) contracted renal arterial rings from both strains; the contraction at the highest concentration was significantly larger in SHR (15±0.7 mN) than in WKY (10±1.5 mN) preparations (
n
=10,
P
<0.05). This phenomenon was blunted by indomethacin, ketanserin, but not S18886. After washout and re-incubation, EDCF responses to acetylcholine (10
-8
to 10
-4
mol/L) were similar between SHR and WKY preparations in the presence of CAY10441 (

E
max

72.6±5.3% and 67.4±3.8% KCl,
n
=8), but significantly larger than under control conditions (
n
=4-8,
P
<0.05). S18886 prevented EDCF responses in the absence but not in the presence of CAY10441. Contractions to prostacyclin [in the presence of indomethacin] were facilitated by CAY10441 in rings from both strains and no longer prevented by S18886, although the TP receptor antagonist abolished contractions to U46619 (10
-11
to 3 x 10
-6
mol/L). These data suggest that: a) inhibitory IP receptors reduce renal vasomotor tone; b) IP and TP receptors form heterodimers; and c) endogenous serotonin contributes to the action of prostacyclin in rat renal arteries.
Abstract 337: G Protein-coupled Estrogen Receptor Contributes To The Vascular Effects Of Aldosterone And Type Two Diabetes-associated Vascular Dysfunction
Authors: Ferreira Nathanne S , Cau Stêfany B , Manzato Carla P , Silva Marcondes A , Carneiro Fernando S , Tostes Rita C
Published: March 2022

Aldosterone (Aldo) excess aggravates endothelial dysfunction in diabetes. Aldo exerts its effects via activation of both mineralocorticoid receptors (MR) and G protein-coupled estrogen receptors (GPER). Considering that GPER activation has beneficial effects in the vasculature, we hypothesized that GPER-mediated vascular effects of aldosterone are decreased/abrogated in diabetes. Second-order mesenteric arteries from control (B6BKS-Leprdb/+) and diabetic (db/db) female mice were incubated with 10 nM Aldo, in the presence of either vehicle (veh), the MR antagonist eplerenone (Eple, 10 μM) or the GPER antagonist G15 (1μM), and the effects on phenylephrine (Phe) vascular reactivity were determined. Aldo increased Phe maximal response (Emax, % of KCl contraction) in arteries from control (veh: 112.5±3.2 vs. Aldo: 129.1±2.8 p0.05). In control vessels, Eple did not alter Phe Emax either in the presence of Aldo or veh (p>0.05), whereas G15 abrogated Aldo-induced increase in Phe Emax (Aldo: 129.1±2.8 vs. G15+Aldo: 110.3±3.6 p<0.05). In db/db arteries, the MR and GPER antagonist decreased Phe Emax both in the presence of veh and Aldo (veh: 143.8±4.9 vs Aldo: 136.0±5.2 vs. Eple+veh: 105.4±4.8 vs. Eple+Aldo: 94.6±4.4 vs. G15+veh: 96.4±4.2 vs. G15+Aldo: 104.3±4.1 p<0.05). Arteries from diabetic mice exhibited increased ERK1/2 activation vs. control [arbitrary units (Au), 229.4±3 vs. 100.0±11.8, respectively, p<0.05]. Aldo increased ERK1/2 activation in control (Au, 99.8±0.4 vs. 272.2±58.0 p<0.05) and db/db mice (Au, 99.7±4.0 vs. 162.4±28.2 p<0.05). Eple and G15 reduced Aldo-induced ERK 1/2 activation in control arteries (p<0.05). In db/db arteries, G15 decreased ERK1/2 activation both in the presence of veh and Aldo (Au, Aldo: 162.4±28.2 vs. G15+veh: 81.1±9.1 vs. G15+aldo: 82.7±7.5 p<0.05). In summary, Aldo acutely increases contractile responses to PhE in mesenteric arteries from control animals by GPER-dependent mechanisms. In diabetic animals, both MR and GPER seem to contribute to Aldo effects in the vasculature. Contrary to our hypothesis, GPER contributes to the vascular effects of Aldo in diabetic animals as well as to diabetes-associated vascular dysfunction.
Abstract 338: 20-HETE Contributes To Ischemia-induced Compensatory Neovascularization
Authors: Chen Li , Zhang Fank F , Gotlinger Katherine H , Falck John C , Scicli Alfonso G , Schwartzman Michal L , Guo Austin M
Published: March 2022

Objective:
Compensatory neovascularizaiton is an important adaptation for recovery from critical ischemia. Recent studies identified the CYP4A-20-HETE system as a novel regulator of angiogenic processes. Here, we assessed the contribution of the 20-HETE system to ischemia-induced neovascularization and further explored its underlying molecular and cellular mechanisms.

Methods:
The mouse ischemia hindlimb angiogenesis assay was performed to evaluate the effect of systemic or local inhibition of the CYP4A-20-HETE system with the 20-HETE synthase inhibitor, DDMS or the 20-HETE antagonist, 20-HEDGE on the compensatory angiogenic responses. Laser Doppler Perfusion Imaging was conducted to assess hindlimb blood flow. Blood pressure was monitored with the non-invasive blood pressure monitoring system. 20-HETE production in ischemic muscles was also measured by LC-MS-MS analysis. IHC staining were carried out to quantify microvessel density and to assess the localization of 20-HETE system in hindlimb gracilis muscle. Finally, western blot was performed to explore the potential underlying signaling pathways.

Results:
Inhibition of 20-HETE synthesis or antagonizing its action either locally or systematically reduced the blood flow perfusion and microvessel formation in response to ischemia without affecting blood pressures. Importantly, ischemic hindlimb muscles showed markedly elevated 20-HETE synthesis compared to non-ischemic controls (96 ± 17 pg/mg of protein versus non-detectable amount). IHC staining showed that an upregulation of 20-HETE synthase (
cyp4a12a
) is co-localized with endothelial cells in the ischemic gracillis muscle, suggesting ischemia may induce the increased production of 20-HETE in the vasculature. Furthermore, the protein expression of HIF-1α, VEGF, VEGFR2, and p44/42 MAPK in ischemic hindlimb muscles were significantly induced. These increases were negated by DDMS or 20-HEDGE.

Conclusions:
Early increases in 20-HETE production in ischemic muscles may regulate ischemic angiogenesis
via
the induction of the HIF-1α/VEGF and MAPK pathways. These results strengthen the notion that 20-HETE may be a key regulator of ischemia-induced compensatory neovascularization processes.
Abstract 342: Vascular Endothelial Ace Deletion Or AT1R Blockade Reduces Blood Pressure But Does Not Prevent Vascular Remodeling In Models Of 20-HETE-Dependent Hypertension
Authors: Joseph Gregory , Garcia Victor , Steadman Elisabeth , Ding Yan , Shkolnik Brian , Capdevila Jorge , Falck John R , Bernstein Kenneth E , Schwartzman Michal L
Published: March 2022

20-HETE-mediated hypertension in rats and mice is associated with increased vascular ACE expression and Ang II levels. We examined the contribution of vascular ACE to blood pressure (BP) and vascular remodeling in 20-HETE-dependent hypertension. The ACE3/3 mice lack vascular endothelial ACE, exhibit attenuated renal ACE expression and display normal plasma ACE and Ang II levels. Administration of 5α-dihydrotestosterone (DHT) (100 mg; 21-day pellet), an inducer of 20-HETE production, increased systolic BP (SBP) by 50% in WT mice (148±4 vs 99±2 mmHg) and 26% in ACE 3/3 mice (125±1 vs 99±1 mmHg). Administration of losartan (10 mg/kg/day) or 20-HEDGE (20-HETE antagonist, 10mg/kg/day) at day 14 of DHT treatment reduced SBP in WT and ACE3/3 mice to levels of untreated mice. DHT increased ACE protein in WT by 2.3-fold in mesenteric and 2.6-fold in aortic arteries while ACE3/3 mice lacked vascular ACE expression which remained absent under DHT treatment. DHT increased preglomerular microvessel (PGMV) remodeling in WT and ACE3/3 mice; however, remodeling was attenuated in ACE3/3 mice (media thickness, 14.54±1.24 vs 9.80±0.56 μm; p<0.05) as opposed to WT mice (22.17±0.92 vs 9.68±0.76 μm; p<0.05). In Cyp4a12tg mice, in which the expression of the Cyp4a12-20-HETE synthase is under the control of the doxycycline (DOX) promoter, DOX (1 mg/ml drinking water) increased SBP (136±2 vs 102±1 mmHg; p<0.05). The increase in SBP was prevented by co-treatment with 20-HEDGE (103±3mmHg), lisinopril (100±3mmHg), or losartan (102±3mmHg). DOX treatment increased PGMV remodeling when compared to untreated Cyp4a12tg mice (media thickness, 16.1±0.9 vs. 8.2±0.5 μm, p<0.05). Co-treatment with 20-HEDGE (10 mg/kg/day) prevented DOX-induced PGMV remodeling (8.8±0.6 μm, p<0.05) while co-treatment with lisinopril or losartan attenuated but did not prevent PGMV remodeling (12.0±0.5 and 11.1±0.5 μm). DOX-treated Cyp4a12tg displayed increased ACE expression in PGMV which was negated by treatment with 20-HEDGE but unaffected by either losartan or lisinopril treatment. These results suggest that vascular endothelial ACE contributes, in part, to 20-HETE-mediated microvascular remodeling in hypertension and that 20-HETE have effects on microvascular remodeling independent of RAS.
Abstract 345: Role Of Vascular Smooth Muscle Cell PPARγ In Aldosterone-induced Vascular Injury
Authors: Ouerd Sofiane , Trindade Michelle , Idris-Khodja Noureddine , Barhoumi Tlili , Offermanns Stefan , Gonzalez Frank J , Paradis Pierre , Schiffrin Ernesto L
Published: March 2022

Background:
Peroxisome proliferator activated receptor γ (PPARγ) agonists improve vascular remodeling and endothelial dysfunction in hypertensive rodents and humans. PPARγ activation in vascular smooth muscle cells (VSMC) may be responsible for the vascular protective effects of PPARγ agonists. We previously observed a protective role of VSMC PPARγ in angiotensin II-induced endothelial dysfunction and vascular remodeling. However, it is unknown whether VSMC PPAR plays a similar protective role in adverse vascular effects of aldosterone. We hypothesized that inactivation of the
Ppar
gene in VSMC (sm
Pparγ
-/-
) would exaggerate aldosterone-induced vascular injury.

Methods:
Using a tamoxifen-inducible Cre/loxP system,
Pparγ
was ablated in VSMC of adult mice. Thirteen week-old control and sm
Pparγ
-/-
mice were infused or not with aldosterone (400 μg/kg/d, SC) for 14 days while receiving 1% NaCl/0.3% KCl in drinking water. Endothelial function and vascular remodeling were assessed in mesenteric arteries (MA) by pressurized myography.

Results:
Endothelium-dependent relaxation (EDR) responses to acetylcholine were reduced to a similar extent in sm
Pparγ
-/-
and aldosterone-treated control and sm
Pparγ
-/-
mice compared to control mice (E
max
: 62.5±8.7%, 50.8±8.6% and 56.8±7.9%, respectively, vs 86.4±3.2%,
P
<0.05). L-NAME, an inhibitor of nitric oxide (NO) synthase, completely blocked EDR in the four groups. Endothelium-independent relaxation response to the NO donor sodium nitroprusside and contractile responses to norepinephrine were similar in the four groups. Preliminary data indicated that aldosterone tended to increase MA stiffness in control mice, as shown by a leftward shift of the stress/strain relationship curve (strain at 140 mmHg, 0.79±0.07 vs 0.89±0.03). Furthermore,
Pparγ
deletion induced an increase in MA stiffness compared to control, which was not worsened by aldosterone (strain at 140 mmHg, 0.67±0.01, 0.65±0.03, vs 0.89±0.03).

Conclusion:
These results indicate that either VMSC
Pparγ
inactivation or aldosterone treatment induce vascular remodeling and endothelial dysfunction, which are not mutually exaggerated. This suggests that PPARγ and aldosterone signal intracellularly through different pathways.
Abstract 351: Activation Of The Central Renin Angiotensin System (RAS) Causes Selective Cerebrovascular Dysfunction
Authors: De Silva T. Michael , Lynch Cynthia M , Grobe Justin L , Faraci Frank M
Published: March 2022

Approximately 30% of hypertensive patients have low plasma renin. The deoxycorticosterone (DOCA)/salt model of hypertension exhibits low peripheral RAS activity, but activation of the central RAS. The impact of central RAS activation on the cerebrovasculature is not defined. We tested the hypothesis that activation of the central RAS would impair endothelial function in brain. We produced mild hypertension in male C57BL/6 mice using subcutaneous DOCA and 0.9% salt in their drinking water for 3 weeks (no nephrectomy was performed) followed by studies of isolated arteries
in vitro
. Blood pressure was elevated modestly after DOCA/salt treatment (79±2 vs 95±3 mmHg). Endothelial function was severely impaired in cerebral arteries from mice treated with DOCA/salt (e.g. 10 μM acetylcholine: 33±3% vs 4±4%) but was unaffected in similar-sized mesenteric arteries. Because microvascular dysfunction is a key feature of small vessel disease in brain that increases the risk of stroke and cognitive impairment, we examined responses of small cerebral arterioles (baseline diameter ~30 μm)
in vivo
using a cranial window. Endothelium-dependent dilation to acetylcholine in cerebral arterioles was reduced by ~75% following DOCA/salt treatment (e.g. 10 μM acetylcholine: 26±5% vs 6±3%). In contrast, endothelium-independent vasodilation to nitroprusside (NO donor) or nifedipine (Ca
2+
channel blocker) was similar between groups. Microvascular endothelial dysfunction was not affected by a scavenger of superoxide but was restored to normal by local inhibition of Rho kinase (ROCK) with Y27632. Activation of AT1 receptors by angiotensin II can increase ROCK signaling. Local inhibition of AT1 receptors with losartan or inhibition of mineralocorticoid receptors (MR) with eplerenone improved endothelium-dependent dilation in DOCA/salt treated mice to normal levels. RhoA and ROCK2 mRNA expression and total endothelial NO synthase protein expression was similar between groups. These findings suggest that activation of the central RAS profoundly but selectively impairs endothelial function in brain. Our findings identify AT1 receptors, MR and ROCK as key contributors to endothelial microvascular dysfunction in this model of mild DOCA/salt-induced hypertension.
Abstract 402: Surgical Treatment Of Aldosterone Producing Adenoma Could Change Renovascular Hemodynamics –Decrement Of Resistive Index Evaluated By Renal Doppler Sonography–
Authors: Iwakura Yoshitsugu , Morimoto Ryo , Kudo Masataka , Ono Yoshikiyo , Ito Sadayoshi , Satoh Fumitoshi
Published: March 2022

Patients with untreated primary aldosteronism (PA) often show albuminuria, which is a predictor for the decrement of their eGFR after specific treatment. Although the appropriate treatment of PA patients results in reducing albuminuria, the mechanism is still unclear. We planned a prospective study to investigate the effects of aldosterone on renal hemodynamics. Renal Doppler sonography (RDS) is recognized as non-invasive device to evaluate renal hemodynamics. Renal resistive index (RI), which is calculated by peak-systolic (PSV) and end-diastolic (EDV) flow velocities (RI = (PSV - EDV) / PSV) measured by RDS, has been reported as a surrogate marker for renal function and tissue injury.

Ninety four patients with aldosterone producing adenoma (APA) were participated; all of those fulfilled diagnostic criteria of PA and underwent adrenalectomy based on results of adrenal venous sampling. RDS was performed at baseline and followed at 1 and 12 months after adrenalectomy. Ninety one patients with essential hypertension were participated in this study as a control. All sonographic parameters were obtained at bilateral renal arteries including main tract, portal, segmental and interlobular parts and expressed as mean. Aldosterone, blood pressure (BP), albuminuria and estimated glomerular filtration rate (eGFR) were followed throughout the study period. Chronic kidney disease (CKD) was defined as eGFR<60 ml/min/1.73m2 or albuminuria≧30 mg/g creatinine.

Initial RI in the patients with untreated APA at all compartment arteries were significantly higher than those in control subjects (0.70 v.s 0.67 at main tract, 0.67 v.s 0.64 at portal part, 0.64 v.s 0.60 at segmental part and 0.63 v.s 0.60 at interlobular part). With significant decrease in aldosterone (from 28.7 to 8.8ng/dl), albuminuria and eGFR significantly declined after adrenalectomy. RI also immediately showed significant declines to 0.67 at main tract, to 0.63 at portal part, to 0.61 at segmental part and to 0.60 at interlobular part, respectively, at 1 month and remained stable through the follow-up period. Furthermore, RI significantly decreased in the APA patients with or without CKD.

Surgical treatment of APA could change renovascular hemodynamics according to the decrease of aldosterone.
Abstract 407: Aminopeptidase A (APA)/Ang(2-10) and APA/AngIII are the Dominant Axes in the Metabolism of Angiotensin Peptides in Mice Plasma
Authors: Wysocki Jan , Haque Syed K , Ye Minghao , Batlle Daniel
Published: March 2022

ACE2 is considered an important enzyme in the degradation of Angiotensin (Ang) I (1-10) and especially of Ang II (1-8), as demonstrated by the rapid dissipation of this peptide when recombinant ACE2 is infused but the steady state level of ACE2 in plasma is low. We examined levels of multiple Ang peptides in plasma under steady state conditions in WT and ACE2KO mice on C57BL genetic background (n=4 and 5, respectively). Liquid chromatography mass-spectrometry-based approach which enables concurrent evaluation of ten different angiotensins at physiologic levels (pg/mL) was used for multiple peptide quantification.

The levels of angiotensin peptides in WT mice were highest for Ang I (1-10) (1295±313 pg/mL), Ang (2-10) (657±173 pg/mL) and Ang III (2-8) (1047±178 pg/mL). In contrast, plasma levels of Ang II (1-8), the main effector peptide of the RAS, and Ang (1-7), the product of cleavage of Ang II by ACE2, were very low (7.8±2 and 3.5±0 pg/mL, respectively). In ACE2 KO mice, Ang II and Ang (1-7) levels were not significantly different from WT. Activity of Aminopeptidase A, an enzyme that converts Ang (1-10) to Ang (2-10) and cleaves Ang II (1-8) to Ang III (2-8), was significantly increased in ACE2KO (n=11) as compared to WT (n=17) (149±11 vs. 94±9 RFU/ul/hr, respectively, p<0.001). This finding along with increased ratio of Ang (2-10)/Ang(1-8) (175±31 vs. 84±4, p<0.01, respectively) suggests increased formation of Ang (2-10) from Ang (1-10) in ACE2KO mice by APA over the formation of Ang II (1-8) from Ang (1-10).

These results show that plasma Ang II and Ang (1-7) levels in WT and ACE2KO mice are low relative to the high levels of Ang (2-10) and Ang III (2-8). ACE2 deficiency does not result in plasma Ang II accumulation or reduced Ang (1-7) levels and is associated with an increase in serum APA activity. We conclude that in mice plasma the APA/Ang(2-10) and APA/AngIII pathways, and not ACE2/Ang(1-7), are dominant in the metabolism of Ang I and Ang II under steady state conditions.
Abstract 413: Hypothalamic Leptin Signaling And Blood Pressure Regulation In Obese Rats
Authors: Silva Carla G , Marin Rodrigo M , Katashima Carlos K , Marcondes Fernanda K , Saad Mario J
Published: March 2022

The main function of Leptin is to inform the individual's nutritional status to hypothalamus, which in turn regulate food intake and energy expenditure. Such actions are orchestrated by phosphorylation of ObRb receptor on tyrosine 985 and 1138, the latter is related to metabolism and Stat3 pathway. Recently, it was demonstrated that high serum levels of leptin, which is seen in DIO, correlates strongly with hypertension. On the other hand, obese individuals have resistance to metabolic actions of leptin (satiety and thermogenesis).

To investigate the molecular hypothalamic pathways that characterize the selective activity of leptin on the effect of blood pressure (BP) increase, adults Sprague Dawley rats were fed with standard diet (SD) or HFD for three months and were submitted to chronic ICV injection of leptin (2μg/day) or saline (2μl) for 10 days. BP, food intake and energy expenditure were assessed before and after treatment.

SD+leptin group had reduced food intake from 109,48±8,48 to 57,3±7,04 kcal / day, increased energy expenditure from 85,6±1,87 to 94,4±0,58 kcal/day/kg^0,75 and consequently weight reduction of 15,25±3,54 g (P <0.001 all) which did not occur in HFD+leptin. However, both SD and HFD group, leptin increased BP of 10.44±0.12% (from 103,7±0,7 to 114,8±0,8 mmHg) and 14.22±1.9% (from 118,6±1,3 to 132,2±1,5 mmHg), respectively.

As expected, obese animals had elevated levels of serum leptin when compared to SD group (SD: 2,24±0,38 ng/dl and HFD: 9,88±1,82 ng/dl, p<0,05). Additionally, we evaluate the main activated signal pathways by leptin in hypothalamus. SD+leptin group showed a significant increase in activity of Stat3, Akt and in MAPK expression. However, HFD+leptin group manifested an increase only on the expression of MAPK pathway. Both SD and HFD leptin-treated groups had higher ObRb tyr985 phosphorylation, which was not observed with the pObRb tyr1138. It was possible to see that ObRb tyr985 dependent pathway is responsive to leptin in SD and HFD, and ObRb tyr1138 as well as the related protein Stat3 and Akt are reduced in obese animals. Therefore, it may be suggested that in HFD-induced obesity hypothalamic pathway ObRb tyr985 / MAPK, unlike the other pathways, is not resistent and can be related to cardiovascular actions of leptin.
Abstract 421: Alterations in Adherens Junction and Gap Junction Precede Desmosomes Remodeling During the Transition from Experimental Compensated Cardiac Hypertrophy to Decompensation
Authors: dos Santos Daniele O , Blefari Valdecir , Prado Fernanda P , Silva Carlos A , Fazan Rubens , Salgado Hélio C , Ramos Simone G , Prado Cibele
Published: March 2022

Hypertension causes cardiac hypertrophy, cardiac dysfunction, and heart failure (HF). The mechanisms implicated in the transition from physiological to pathological hypertrophy are not fully understood. Growing evidence points out to the role of inappropriate intercellular mechanical and electrical coupling in the pathogenesis of HF. We investigated adherens junction, desmosome and gap junction proteins and their potential role in the pathogenesis of HF in rats submitted to chronic abdominal aorta stenosis. Wistar rats were submitted to abdominal aorta stenosis and killed at 90 days post surgery: 60% presented hypertrophied hearts (HH) and 40% hypertrophied and dilated hearts (DH), as evaluated by echocardiography. Blood pressure was evaluated. The hearts were collected, Western blot (WB) and immunofluorescence (IF) were performed for cadherin, plakoglobin, B-catenin, desmoglein, desmocollin and connexin-43. Cardiac systolic function was evaluated using Vevo 2100 ultrasound system. Data were considered significant when p<0.05. Cadherin expression increased 55% in HH and decreased 33% in DH; plakoglobin increased 67% in HH and decreased 29% in DH; B-catenin increased 73% in HH and decreased 32% in DH; connexin-43 decreased 17% in HH and 32% in DH; desmocolin increased 57% only in HH group and no alteration in desmoglein expression was observed in both groups. Mean blood pressure increased 61% in HH and 59% in DH. Ejection fraction and fractional shortening decreased 45% and 41%, respectively, only in DH. These results suggest different sensitivity of the intercalated disc components: proteins of the adherens junction and gap junction are more sensitive than desmosomes proteins and present significant reduced expression. This reduced expression could contribute to the transition of the compensated cardiac hypertrophy to decompensation. These abnormal parameters may emerge as therapeutic targets to modulate the evolution of HF.
Abstract 425: The Effectiveness Of Metoprolol Versus Atenolol On Prevention Of All-cause And Cardiovascular Mortality In A Large Chinese Population: A Cohort Study
Authors: Wong Martin C , Tam Wilson W , Lao Xiang Qian , Wang Harry H , Kwan Mandy W , Cheung Clement S , Tong Ellen L , Cheung Ngai Tseung , Yan Bryan P , Yu Cheuk Man , Griffiths Sian M
Published: March 2022

Background:
Existing trials almost exclusively used atenolol to represent the entire beta-blocker class. We compared the incidence of all-cause and cardiovascular (CVS) mortality, blood pressure (BP) control and adherence levels between patients newly prescribed atenolol vs. metoprolol.

Methods:
This cohort study included all public, clinical settings in Hong Kong between 2001 and 2005, followed up till 2010. We compared outcomes between 22,479 new atenolol users and 29,972 new metoprolol users. The difference in mortality between drugs was evaluated by Cox proportional hazard regression models. Binary logistic regression analyses were used to compare the BP control rates and adherence levels.

Results:
Among atenolol and metoprolol users, 7.0% (1,577 of 22,479) and 13.1% (3,940 of 29,972) died, respectively (p<0.005). The incidence of CVS mortality among atenolol users was lower than metoprolol users (1.4% vs. 3.7%, p<0.001). When compared with atenolol users, metoprolol users were 1.13-fold (95% C.I. 1.06-1.20) and 1.56-fold (95% C.I. 1.27-1.90), respectively, more likely to encounter all-cause and CVS mortality; less likely to be drug adherent (Adjusted Relative Risk [RR]: 0.95, 95% C.I.: 0.90-0.99, p=0.013); and less likely to achieve optimal overall BP control (RR 0.94, 95% C.I. 0.90-0.99, p=0.023) and diastolic BP control (RR 0.86, 95% C.I. 0.77-0.97, p=0.013).

Conclusions:
These findings imply an intra-class difference for beta-blockers when used as first-line antihypertensive prescriptions in real-life clinical settings. More outcome studies on the effectiveness of different subtypes within other major antihypertensive drug classes are warranted.
Abstract 430: Impairment of Instant Membrane Resealing by Lipid Raft Clustering in Endothelial Cells: Role in NLRP3 Inflammmasome Activation
Authors: Xia Min , Chen Yang , Yuan Ming , Li Pin-Lan
Published: March 2022

Instant membrane resealing importantly contributes to the functional and structural integrity of the endothelial cells (ECs) that are exposed to various physical and chemical stimuli in blood stream. The present study was designed to explore the molecular mechanisms mediating this endothelial membrane resealing with a focus on the role of lipid rafts (LR) clustering. Using high energy Laser gun, a tiny hole was made in cultured EC bathed with FM1-43, and the rapid entry of this FM1-43 to produce fluorescence was used to measure membrane resealing. We demonstrated that ECs exhibited a Ca2+-dependent instant membrane resealing, as shown by a significant reduction of fluorescence appearance within these cells compared to ECs bathed with Ca2+ free solution. This Ca2+-dependent instant membrane resealing was also observed in ECs up stimulation of Lactobacillus casei cell wall fragments (LCWE), which was commonly used to produce arteritis. When ECs were pretreated with LRs clustering stimulators such as C-16 ceramide, ASMase or FasL, EC membrane resealing was substantially abolished and membrane wounding areas are enriched with LR clusters. By dual wavelength high speed wavelength switching microscopy, we found that a small Ca2+ influx (fura-2 fluorescence) was recorded at beginning of LCWE treatment of ECs (about 10 min), but membrane was immediately resealed (no PI fluorescence). However, if the LR stimulator FasL was used to pretreat these ECs, a large flushing of both PI and Ca2+ into ECs was monitored. By calculation of df/dt and time of PI fluorescence reaching the maximum, it was shown that FasL largely increased LCWE-induced PI fluorescence to a level equal to ECs with Ca2+-free solution over a 20-min period (maximum df/dt=1.17 fold/sec±0.22 at 14 min with FasL+Ca2+, 0.87 fold/sec±0.21 at 19 min with Ca2+ free, and 0.03 fold/sec±0.01 at 20 min with Ca2+ in bath). In addition, it was shown that impaired EC membrane resealing significantly enhanced the production of active caspase-1, a marker of NLRP3 inflammasome activation (a ~3-fold increase). These results strongly suggest that ceramide-associated LR clustering impairs the instant membrane resealing in ECs, which may activate EC inflammasomes leading to vascular inflammation and injury.
Abstract 432: Comparison of Body Mass Index with Waist Circumference, Waist-to-height Ratio and Waist-to-hip Ratio as a Predictor of Incident Hypertension in Japan
Authors: Nakai Michikazu , Watanabe Makoto , Nishimura Kunihiro , Higashiyama Aya , Takegami Misa , Kokubo Yoshihiro , Nakao Yuko M , Kobayashi Takashi , Watanabe Takuya , Okamura Tomonori , Miyamoto Yoshihiro
Published: March 2022

Objective:
Obesity is an established risk factor for hypertension (HT). However, there are few studies to investigate which obesity-related indicator has better predictability for HT in Asian general population. We performed a population-based prospective cohort study in Japan to compare the predictability of four indicators (body mass index (BMI), waist circumference (WC), waist-to-height ratio (WHtR) and waist-to-hip ratio (WHR)) for HT.

Methods:
Participants were 1,591 men and 1,973 women aged 30-84 years without HT at baseline. BMI(weight/height2), WC, WHtR(WC/height) and WHR(WC/Hip Circumference) were measured at baseline, and participants were followed for incidence of HT. The Cox proportional hazards model was used to estimate hazard ratios (HRs) of each indicator for incident HT by sex after adjusting for age, cigarette smoking and alcohol drinking. The paired homogeneity test was conducted to observe statistical differences of predictability between BMI and other indicators.

Results:
During mean follow-up of 7.2 years, 640 men and 685 women developed HT. HRs (95% CI) of BMI, WC, WHtR and WHR for incident HT by a 1 SD increase were 1.25(1.15,1.35), 1.21(1.12,1.31), 1.23(1.14,1.34) and 1.14(1.06,1.24) in men while 1.32(1.22,1.42), 1.27(1.18,1.37), 1.32(1.21,1.44) and 1.22(1.12,1.32) in women, respectively. There was no statistical difference of the predictability between BMI and the two indicators (WC and WHtR) in both sexes, but WHR was inferior to BMI with significance in men (p=0.02) or marginally significance in women (p=0.06).

Conclusion:
All four obesity-related indicators could predict incident HT in a Japanese population. BMI, WC and WHtR were similar in the predictability, but WHR was a little inferior to BMI.
Abstract 439: Gender and Racial Trends in Blood Pressure Control in a University-Based Cardiology Clinic
Authors: Ul Haq Ehtesham , Omar Mazen , Omar Bassam A
Published: March 2022

Background:
Gender and racial differences may influence control of coronary disease risk factors, and have been partially attributed to socioeconomic issues.

Methods:
Encounters from a university-based primary cardiology clinic with broad referral base were analyzed for BP by gender and race in patients with hypertension as a diagnosis.

Results:
200 encounters were analyzed: 105 (53%) females; systolic BP (SBP) 134 ± 20 mmHg compared with 95 (47%) males; SBP 130 ± 18 mmHg (NS). There were 100 (50%) African American patients; SBP 134 ± 20 mmHg compared with 100 (50%) White patients; SBP 130 ± 18 mmHg (NS). Comparing each subgroup to the remainder of the patients revealed African American females (AAF) with SBP of 137 ± 21 mmHg (P < 0.01), White females with SBP 129 ± 19 mmHg (NS), African American males with SBP 128 ± 18 mmHg (NS), White males SBP 131 ± 18 mmHg (NS). Diastolic BP was 83 ± 13 mmHg in AAF compared with 78 ± 11 mmHg in the remaining groups (P < 0.01). Age, heart rate, LDL levels and weights in AAF were 59 ± 13 years, 70 ± 10 BPM, 91 ± 44 mg/dL and 214 ± 77 lbs compared with 61 ± 13 years, 70 ± 12 BPM, 88 ± 38 mg/dL and 209 ± 68 lbs respectively in the remaining patients (NS).

Conclusion:
African American females with treated hypertension had the highest documented BP during their clinic visits. Lack of compliance with medications is unlikely because these patients had insurance and their LDL levels were adequately controlled with medications. Body weight, though not statistically significant, may have contributed to the elevated BP in AAF. White-coat-hypertension is another potential explanation, which has been previously reported to be prevalent in African American females. This is an important observation given that approximately 50% of AAF patient in our cohort had treated diabetes requiring stricter BP control, compared with 25% in the other groups. Further studies with ambulatory BP monitoring may help better explain this observation. AAF remain at exceptional risk for cardiovascular disease; efforts to improve detection and control of hypertension, in addition to other cardiac risk factors, in this vulnerable ethnic/gender segment of the population are of paramount importance.
Abstract 464: Effects of Increased Potassium and Sodium on Endothelial Function: A Pilot Study
Authors: Blanch Natalie , Clifton Peter M , Keogh Jennifer B
Published: March 2022

Background and aim:
Increased potassium intake has been related to reduced blood pressure (BP) and improved endothelial function. High sodium intake increases BP and impairs endothelial function. The effect of increasing potassium in the presence of high dietary sodium is not known. The aim was to determine the effect of increased potassium and increased sodium on post prandial endothelial function, as assessed by flow mediated dilatation (FMD).

Materials and methods:
Eighteen healthy, normotensive volunteers (age 41±16 and BMI 23.0±2.7) received a meal with 3.1mmol potassium and 65mmol sodium (LKHNa), a meal with 38mmol potassium and 65mmol sodium (HKHNa) and a control meal (LKLNa) with 5.5mmol sodium and 3.1mmol potassium on three separate occasions in a randomized order. FMD, pulse wave velocity (PWV), augmentation index (AI) and BP were measured while participants were fasting and at 30, 60, 90 and 120 minutes after the meal.

Results:
The LKHNa meal produced a significant reduction in FMD at 90 minutes when compared to the LKLNa control meal (p<0.01). The addition of potassium (HKHNa) significantly attenuated the post meal decrease in FMD when compared to the high sodium meal (p<0.05) at 90 minutes (Figure 1) . There was no significant differences in PWV, AI, or BP. The study will continue until n=40.

Conclusions:
The addition of potassium to a high sodium meal may attenuate the post meal reduction in endothelial function as assessed by FMD. Increases in sodium and potassium do not appear to effect PWV, AI or BP in the postprandial state. Additional data collection will continue to elucidate the effect of increased potassium in the presence of high sodium.
Abstract 465: AT
2
Receptor Signaling Plays An Important Role In Conversion Of From White To Beige Fat
Authors: Tsukuda Kana , Mogi Masaki , Nakaoka Hirotomo , Kan-no Harumi , Chisaka Toshiyuki , Wang Xial-Li , Kukida Masayoshi , Bai Hui-Yu , Shan Bao-Shuai , Iwanami Jun , Horiuchi Masatsugu
Published: March 2022

Background:
We have previously reported that treatment with angiotensin II type-2 (AT
2
) receptor agonist (compound 21; C21) ameliorated insulin resistance with the improvement of adipocyte dysfunction in obese type 2 diabetic mouse model, KKAy. Until quite recently, adipose tissue has been divided into two groups that have directory-opposed capacities; white adipose tissue is best known for its role in energy store, while brown adipose tissue functions for energy expenditure (thermogenesis). In addition, new clusters of adipocyte with thermogenic capability have been discovered in white adipose tissue, which are distinct origins from brown adipose tissue. These adipocytes have been named ‘beige’. However, the role of AT
2
receptor signaling in white-to-beige fat conversion is not known.

Methods:
KKAy mice were subjected to i.p. injection of C21 (10ug/kg/day) for 2 weeks. After the treatment, the expression of thermogenic genes (Ucp1, Cidea, Pgc1α) in adipose tissue were determined with real-time RT-PCR. Next, we examined the difference in conversion from white to beige fat between wild-type (WT) mice and AT
2
receptor knockout (AT
2
KO) mice. Eight-week-old mice were fed a high-fat and high-cholesterol diet (HFCD) (10% coconut oil + 1.25% cholesterol) for 2 weeks. To examine the effect of exercise on fat conversion, 9-week-old mice were forced swim test (2 minх3 times/day) for 7days. After the swim test, mice were sacrificed for examinations.

Results:
In KKAy mice, treatment of C21 increased the expression of thermogenic genes in subcutaneous fat, but not in visceral fat. Adipose tissue weight was not changed by treatment of C21. In AT
2
KO mice fed HFCD for 2 weeks, visceral fat was greater than that in WT mice, but subcutaneous fat was not. The expression of thermogenic genes in subcutaneous fat was lower in AT
2
KO mice than in WT mice. In both mice, swim exercise increased the expression of thermogenic genes; however, the up-regulation of these genes in AT
2
KO mice did not reach the level of that in WT mice.

Conclusion:
These results suggest that AT
2
receptor signaling has an important role in white-to-beige fat conversion, and therefore AT
2
receptor signaling might be a potential therapeutic target for lipid metabolic disorders.
Abstract 468: Diversity of Apolipoprotein E genetic polymorphism significance on cardiovascular risk is determined by the presence of Metabolic Syndrome among hypertensive patients
Authors: Teixeira Andrei A , Marrocos Mauro S , Quinto Beata M , Rodrigues Cassio J , Dalboni Maria A , Batista Marcelo C
Published: March 2022

INTRODUCTION:
Hypertension has a significant relevance as a cardiovascular risk factor. A consistent increase on world’s Metabolic syndrome (MetS) incidence has been associated with an epidemic cardiovascular risk in different populations. Dislipidemia plays a major role determining the epidemic CV burden attributed to MetS. Apolipoprotein E (ApoE) is involved on cholesterol and triglycerides metabolism regulation. Once ApoE polymorphism may influence lipid metabolism, it is possible that it brings on individual susceptibility consequences for the development of MetS and cardiovascular risk.

OBJECTIVE:
To measure the discriminatory power of ApoE polymorphism in determining the cardiovascular risk stratification based on the presence MetS in a cohort of hypertensive patients.

PATIENTS AND METHODS:
It was enrolled 401 patients, divided in two groups, classified by MetS presence (IDF criteria): Group 1: 275 patients with MetS (MetS +) and Group 2: 126 patients without Mets (MetS -). Patient’s data were collected by clinical evaluation, physical exam, file reviews and laboratory testing. Polymorphic ApoE analysis was performed by PCR amplification. Groups were compared on clinical and laboratorial characteristics as well as allele and genotype distribution in relation to ApoE polymorphism. Prevalence of CVD, in relation to Mets presence, was analysed based on allele 4 prevalence.

RESULTS:
The results evidenced 186 men (47,4%), 62,1% whites, 44,1% diabetics and 11,5% of patients were smokers. Mean age was 64,0±12,0 years. When genotypic distribution was analyzed, E3/3 genotype and E3 allele frequency were more prevalent. Among patients with MetS, we observed an independent association between CVD prevalence and allele E4 frequency(OR 2.42(1.17-1.5, p<0,05)). On the contrary, in those without MetS, there was lesser CVD burden in E4 allele carriers(OR 0,14(0,02-0,75)). These associations remained significant even after confounding factor corrections.

CONCLUSION:
The results presented show that association between ApoE gene and CVD maybe modulated by de presence of MetS, with an increased CV burden observed among E4 carriers with the Syndrome. On the opposite way, E4 carriers without visceral obesity had a lesser prevalence of CVD.
Abstract 477: Human Ang-(1-12) and Hemodynamic and Cardiac biometrics in Rats Overexpressing Human Angiotensinogen
Authors: Varagic Jasmina , VonCannon Jessica L , Nagata Sayaka , Ahmad Sarfaraz , Ferrario Carlos M
Published: March 2022

It has been reported that insertion of the human angiotensinogen (AGT) gene in rats does not cause hypertension as rat renin has no hydrolytic activity on human AGT. Initial studies in transgenic rats expressing the human AGT gene [TGR(hAGT)L1623] demonstrated the presence of human angiotensin-(1-12) [Ang-(1-12)] immunoreactivity in the heart of these rats. The exciting possibility that [TGR(hAGT)L1623] rats have increased precursor availability for Ang II formation from both human and rat origin stimulated further investigation of this model through measurements of arterial pressure and heart rate with implanted telemetry probes (Data Science International, MN), assessment of cardiac structure and function by echocardiography (VevoLAZR; VisualSonics Inc., Canada), and heart weight (n=9). Comparative studies were done in Sprague Dawley rats (SD; n=11). 24 h mean arterial pressures (Figure), but not heart rates, were significantly higher in [TGR(hAGT)L1623] rats compared to SD controls during 4 weeks of continuous measurement (starting at 13 weeks of age). Although there was no difference in systolic or diastolic function between these two strains, [TGR(hAGT)L1623] rats showed increased thickness of left ventricular posterior wall (2.3 ± 0.1 vs. 2.1 ± 0.1 mm; p<0.05) and diameter (8.1 ± 0.2 vs. 7.5 ± 0.1 mm; p<0.05) that was associated with increased heart weight (1.51 ± 0.03 vs 1.16 ± 0.01 g; p<0.001) and heart weight/tibia length ratio (34.6 ± 0.7 vs 28.6 ± 0.4 mg/mm; p<0.001). Detection of hypertension and cardiac Ang-(1-12) in rats with overexpression of the human AGT gene suggests a non-renin dependent enzymatic mechanism able to cleave Ang-(1-12) from the human substrate.
Abstract 479: Ex vivo Equilibrium Analysis of the Renin-Angiotensin-System: Clinical Implications for Diagnosis and Treatment of Hypertension
Authors: Domenig Oliver , Schwager Cornelia , van Oyen Dunja , Poglitsch Marko
Published: March 2022

The concentrations of angiotensins are maintained by equal rates of formation and degradation. The resulting steady-state angiotensin levels are affected by local molecular factors including tissue, endothelium or blood cell associated angiotensin receptors and processing enzymes as well as plasma soluble RAS components. The high concentration of the pro-hormone angiotensinogen in human plasma in combination with reported ranges for plasma renin activity result in a long-lasting and stable Ang I formation rate without significantly reducing angiotensinogen levels within several hours. This phenomenon can be utilized for the generation of an ex vivo situation that is characterized by significantly higher but stable angiotensin peptide levels. These ex vivo equilibrium angiotensin levels provide an integrated picture about plasma angiotensinase activities and therefore represent a powerful diagnostic tool for analyzing the systemic RAS in clinical samples. Moreover, these ex vivo levels remain stable over hours of incubation at 37°C and show a very high correlation (>90%) with angiotensin levels obtained by a state-of-the art sample collection procedure using complex inhibitor cocktails for stabilizing angiotensin peptides during blood collection.

The quantification of equilibrium angiotensin levels does not require any special sample collection procedures and can be applied to frozen serum and heparin plasma samples. It turned out that ex vivo angiotensin levels represent a measure for the consumption of angiotensin metabolites by the organism, therefore providing a powerful and versatile tool for assessing in vivo angiotensin signaling on the patient specific level enabling new diagnosis based rationales for anti-hypertensive therapies. The application of ex vivo RAS-Fingerprinting in clinical studies could substantially enhance our understanding of the regulation and physiology of the human RAS and could further lead to the development of powerful personalized approaches in the future treatment of hypertension.
Abstract 487: Cardiac Sympathetic Afferent Denervation Improves Cardiac Inflammation and Ameliorates Cardiac Remodeling in Post-MI Rats
Authors: Wang Hanjun , Mishra Paras K , Nandi Shyam S , Cornish Kurtis G , Zucker Irving H
Published: March 2022

A recent study from our laboratory demonstrated that chronic and

selective cardiac sympathetic afferent denervation at the time of myocardial infarction (MI) by epicardial application of resiniferatoxin (RTX), a neuronal toxin capable of inducing rapid degeneration of transient receptor potential vanilloid 1 (TRPV1)-expressing afferent neurons and fibers, largely prevented the deleterious cardiac remodeling process 9-11 weeks post-MI. This included reduced cardiac hypertrophy, fibrosis and apoptosis. Here, we further investigated the effect of epicardial application of RTX during MI on cardiac extracellular matrix (ECM) remodeling at 10 weeks post-MI. Echocardiographic and morphologic data demonstrated that, compared to MI+vehicle, MI+RTX exhibited a significantly slower LV chamber dilation (6-week echocardiographic data: left ventricular end-diastolic diameter, 10.7 ± 0.2 vs. 9.6 ± 0.3 mm; left ventricular end-systolic diameter: 8.8 ± 0.2 vs. 7.8 ± 0.3 mm; MI+vehicle vs. MI+RTX, n=18, P<0.05). To further explore its molecular mechanism, we used zymography to evaluate the effect of RTX application on the matrix metalloproteinase (MMP) activity, which is responsible for degrading ECM and cardiac dilation after MI. Our preliminary data show that MMP9 activity was increased by 3.9 times in the peri-infarct myocardium post-MI compared to sham. This was largely abolished by RTX application (peri-infarct area: 3.9 ± 0.5 vs. 1.7 ± 0.4, MI+vehicle vs. MI+RTX, n=3, p=0.03). We further investigated the effect of RTX application on MI-induced cardiac inflammation, which has been reported as an upstream mechanism triggering MMP activation post-MI. Our data show that cardiac TNF alpha levels are markedly increased in the peri-infarct or remote myocardium of post-MI rats, which was largely abolished by RTX application during MI (TNF alpha levels in peri-infarct area: 214.2 ± 13.1 vs. 647.5 ± 45.1 vs. 357.3 ± 25.4 pg/mg protein, sham+vehicle vs. MI+vehicle vs. MI+RTX, n=5/each, p<0.05). These data suggest that cardiac sympathetic afferent denervation during MI exerts a local anti-inflammatory effect and reduces the ECM remodeling by preventing excessive MMP activation in post-MI rats.