bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P211: Intra-abdominal Lipectomy Normalizes Arterial Stiffness and Blood Pressure via Reduction in 20-HETE
Authors: Soler Amanda A , Hutcheson Brenda , Yang Jenny , Bradford Chastity , Zhang Frank F , Gotlinger Katie H , Schwartzman Michal L , Rocic Petra
Published: July 2021

Increased intra-abdominal (visceral) adipose tissue is a key feature of the metabolic syndrome affecting over 30% of the U.S. population. Expansion of visceral adipose tissue is linked to the development of hypertension and is a risk factor for cardiovascular disease that can ultimately lead to end-organ damage. While reduction in visceral adipose tissue volume offers cardioprotective effects, the cardiovascular mechanisms behind these beneficial effects remain unclear. In this study, we removed ~90% of visceral adipose tissue (=~5% body weight) by intra-abdominal lipectomy and assessed large arterial stiffness, large artery structural matrix components, and blood pressure in a metabolic syndrome rat model (JCR:LA-cp, JCR). Large artery stiffness was significantly elevated in JCR vs. normal (Sprague Dawley, SD) rats (75±2% JCR vs. SD (carotid)) with a concomitant significant increase in MMP12-dependent elastin degradation (3-6 fold vs. SD). Intra-abdominal lipectomy normalized large artery stiffness, blocked MMP12 activation and reduced elastin degradation in JCR animals (~75% (carotid) vs. untreated JCR). Likewise, hypertension in JCR animals was significantly attenuated by intra-abdominal lipectomy (MABP=156±3 mmHg JCR vs. 90±6 mmHg SD vs. 132±4 mmHg JCR+lipectomy). 20-hydroxyeicosatetraeonic acid (20-HETE), an arachidonic acid metabolite known to be a potent vasoconstrictor in resistance arteries, was significantly elevated in the visceral adipose tissue of JCR rats (~6 fold vs. SD). Intra-abdominal lipectomy normalized 20-HETE levels in JCR rats. Like intra-abdominal lipectomy, 20-HETE antagonists restored large artery elasticity, blocked MMP12 activation and elastin degradation, and significantly decreased blood pressure (125±3 mmHg JCR+20-HETE antagonists) in JCR rats. Thus, 20-HETE may be an important adipokine that mediates the adverse effects of expanded visceral fat volume in the metabolic syndrome and its inhibition may provide a pharmacological approach for the management of central obesity-driven large artery stiffness and hypertension.
Abstract P216: Nicotinamide Nucleotide Transhydrogenase Regulates Mitochondrial Redox Balance and Endothelial Function in Response to Angiotensin II
Authors: Vetkoetter Matthew , Rao K N Shashanka , Shen Xinggui , Pardue Sibile , Krzywanski David M
Published: July 2021

Hypertension (HTN) involves increased peripheral resistance to blood flow mediated by changes in vascular tone. Endothelial dysfunction is an initiating step in HTN and mitochondrial reactive oxygen species (ROS) contribute to endothelial dysfunction. Nicotinamide nucleotide transhydrogenase (NNT) is a critical enzyme in the regulation of mitochondrial NADPH levels and can impact mitochondrial redox status. We have demonstrated that the loss of NNT in C57Bl/6J mice is associated with a more severe hypertensive phenotype in response to Ang II characterized by mitochondrial and vascular dysfunction driven by vascular ROS production
in
vivo
and
in
vitro
. Little is known regarding the role of NNT human endothelial cells, but based on previous studies we hypothesize that the loss of NNT promotes mitochondrial and cellular ROS production and disrupts normal endothelial
•
NO function. In human aortic endothelial cells (HAEC) treatment with 500nM Ang II for two hours significantly increased NNT expression (225
+
1
3%) and glutathione peroxidase 2 (Gpx2) activity (17.81
+
0
.75 vs 29.06
+
2
.53 nmol/min/ug protein) consistent with NNT’s role in driving the mitochondrial antioxidant systems. Conversely, siRNA knockdown of NNT inhibited Gpx2 activity (18.32
+
1.17 vs 13.16
+
0
.7 nmol/min/ug protein) and increased mitochondrial superoxide production (0.12
+
0
.01 vs 0.17
+
0
.01 nmol/mg protein) in response to Ang II. These changes in the redox balance of the mitochondria contribute to mitochondrial dysfunction where the loss of NNT led to a marked decrease in the respiratory control ratio (RCR) of the mitochondria (6.42
+
0
.17 vs 3.83
+
0
.1) of Ang II treated cells. Ultimately, this shift to a pro-oxidative mitochondrial redox phenotype in these cells conspired to inhibit Ang II-induced
•
NO production (35.60
+ 1
.28 vs 30.10
+ 1
.24 pmol/mg protein). Our data indicate that the absence of NNT expression increases the steady state levels of mitochondrial ROS through increased ROS production and decreased antioxidant activity, which is contributes to reduced
•
NO production and endothelial dysfunction. These studies highlight NNT as a novel regulator of mitochondrial redox tone that may critically impact the ability of vascular ROS to contribute to the development of HTN.
Abstract P219: Vascular Cross-talk Between Redox and Calcium Signaling in Hypertension Involves Transient Receptor Potential Melastatin 2 Channel Activation
Authors: Alves-Lopes Rhéure , Montezano Augusto C , Neves Karla B , Anagnostopoulou Aikaterini , Lacchini Silvia , Touyz Rhian M
Published: July 2021

The transient receptor potential melastatin 2 cation channel (TRPM2) is redox-sensitive and promotes Ca
2+
influx after H
2
O
2
activation through oxidative modification and PARP-ADPR-dependent mechanisms. TRPM2 also regulates Na
+
influx, and by increasing [Na
+
]i interferes with the Na
+
-Ca
2+
exchanger (NCX) inducing reverse mode action, promoting Ca
2+
influx. These processes may be driven by Nox4-derived H
2
O
2.
We tested the hypothesis that vascular dysfunction in hypertension involves oxidative stress-induced TRPM2 activation through H
2
O
2
production, which in turn promotes Ca
2+
influx. Mesenteric arteries isolated from wildtype (WT), LinA3 (mice expressing human renin with Ang II-dependent hypertension), Nox4
-/-
and LinA3/Nox4
-/-
mice and vascular smooth muscle cells (VSMCs) from hypertensive and normotensive patients were used. Arteries from hypertensive LinA3 mice, exhibit increased U46619-induced vasoconstriction versus WT mice (Emax - LinA3
vs
WT: 9.37 ± 0.51
vs
6.79 ± 0.29), an effect attenuated by olaparib (PARP-ADPR inhibitor) and 2-APB (TRPM2 blocker) and also increased mRNA expression (Fold change - related to control) of NOX4 (3.05 ± 0.30), TRPM2 (1.38 ± 0.24), NCX (1.973 ± 0.34) and salt inducible kinase 1 (1.833 ± 0.12) and sodium-potassium pump (1.43 ± 0.16), which are activated when intracellular levels of Na
+
rise beyond a critical point. These events seem to be regulated by NOX4, since they were not observed in mesenteric arteries from LinA3/Nox4
-/-
mice. Ang II-induced Ca
2+
influx is potentiated in VSMCs from hypertensive patients (AUC-Ex490/Em535: normotensive: 15400±917.5
vs
hypertensive - 22460±2388), a response followed by increased generation of O
2
-
and H
2
O
2
in cells from hypertensive patients. These ROS effects were attenuated by catalase, and 2-APB, 8-br and olaparib (TRPM2 inhibitors) and benzamil, KB-R7943 and YM244769 (NCX inhibitors). Our data indicate that TRPM2 ion channel activation contributes to redox-sensitive vascular dysfunction in hypertension. These findings suggest that dysregulation of TRPM2-NOX4-derived ROS and NCX may contribute to redox- and Ca
2+
signalling important in vascular function in hypertension. TRPM2 may be a point of cross-talk between ROS and Ca
2+
signalling.
Abstract P224: β-arrestin2 Stimulates SERCA2a SUMOylation and Activity in Cardiac Myocytes to Promote Contractility
Authors: McCrink Katie A , Maning Jennifer , Brill Ava , Vu Angela , Koch Walter J , Lymperopoulos Anastasios
Published: July 2021

Background:
Heart failure (HF) is the most lethal disease worldwide and new treatments are needed. The β
1
-adrenergic receptor (AR) mediates the positive inotropy of catecholamines, partly via Sarco(Endo)plasmic Reticulum Ca
2+
-ATPase (SERCA)-2a activation. Agonist-activated β
1
ARs, however, are desensitized/downregulated in human HF due to the actions of the βarrestins (βarrestin1 and 2). βarrestins are GPCR adapter proteins and signal transducers. βarrestin1 is by far the predominant isoform in the heart and reduces contractility by desensitizing the β
1
AR, whereas βarrestin2 is expressed at negligible levels and is beneficial post-myocardial infarction (MI), as it combats inflammation and apoptosis. Herein, we sought to investigate whether cardiac βarrestin2 exerts any inotropic effects.

Methods:
We used βarrestin knockout (KO) mouse hearts and also performed intra-cardiac adenoviral gene transfer of βarrestin2 (Adβarrestin2) in post-MI mice in vivo. For mechanistic signaling studies we used the cardiomyocyte cell line H9c2.

Results:
SERCA2a SUMO (small ubiquitin-like modifier)ylation and activity and, consequently, cardiac contractility were increased in βarrestin1 KO`s vs. WT`s post-MI. The opposite was true for βarrestin2 KO`s post-MI. Additionally, βarrestin2, but not βarrestin1, was found to directly bind SERCA2a and induce its SUMOylation and activation in mouse hearts in vivo, as well as in cardiomyocytes in vitro acutely in response to β
1
AR stimulation. Interestingly, βarrestin2 did not affect the classic β
1
AR cAMP-dependent pro-contractile signaling pathway in cardiomyocytes, again contrary to βarrestin1. Importantly, and consistent with these findings, Adβarrestin2 gene transfer in post-MI mouse hearts in vivo resulted in enhanced cardiac function (post-MI ejection fraction of Adβarrestin2 vs. control (AdGFP) mice: 40.3
+
1.3% vs. 23.1
+
1.2%, respectively, p<0.05, n=5).

Conclusions:
Cardiac β
1
AR-activated βarrestin2, but not βarrestin1, promotes SERCA2a SUMOylation and activity, increasing cardiac contractility. Given also its anti-inflammatory and anti-apoptotic effects post-MI, cardiac-specific βarrestin2 gene transfer may be a novel and safe inotropic therapy for both acute and chronic HF.
Abstract P236: Site-1 Protease-derived Soluble (Pro) Renin Receptor Mediates Angiotensin II-induced Hypertension via Activation of Intrarenal Renin System
Authors: Peng Kexin , Wang Fei , Xu Chuanming , Lu Xiaohan , Yang Tianxin
Published: July 2021

We have previously shown that collecting duct (CD)-specific deletion of (pro)renin receptor (PRR) attenuates angiotensin II (Ang II)-induced hypertension, accompanied with reduced soluble PRR (sPRR) that exerts antidiuretic action. Recent preliminary and published results demonstrated site-1portease (S1P) but not furin or ADMA19 as the predominant PPR cleavage enzyme. In the present study, we evaluated involvement of S1P-derived sPRR in Ang II-induced hypertension. By radiotelemetry, CD PRR KO mice exhibited reduced MAP on day 7 of Ang II infusion at 300 ng/kg/min as compared with floxed mice (MAP: 118±5 vs. 137±3 mmHg, N=5, p<0.05). Administration of sPRR-His, a histidine-tagged sPRR, at 120 μg/kg/d via i.v. infusion to CD PRR KO mice for additional 7 days largely restored the sensitivity to Ang II (MAP: 139±6 mmHg in sPRR-His +Ang II group vs. 116±5 mmHg in Ang II group, N = 4, p<0.05). The i.v. infusion was achieved via placement of a catheter in jugular vein with the other end connected to mimipump. In C57/BL6 mice, administration of a S1P inhibitor PF429242 (PF) via mini pump infusion at 30 mg/kg/d for 7 days attenuated Ang II-induced increases in MAP (day 7: 125±5 in Ang II+ PF group vs. 142±3 in Ang II group; N=6, p<0.05), urinary sPRR excretion (27±4 vs. 63±9 pg/24h; N=6, p<0.05). In parallel, urinary renin levels were elevated by Ang II, which was blunted by PF (renin activity: 0.17±0.03 in Ang II+PF vs. 0.80±0.081 in Ang II vs. 0.12±0.016 ng/24h in Control, N=6, p<0.01; active renin content: 15.2±2.7 vs. 236.0±23.2 vs. 5.6±1.3 ng/24h, N=6, p<0.01; prorenin content: 9.6±3.1 vs. 27.8±6.1 vs. 6.2±1.8 ng/24h, N=6, p<0.05; total renin content: 24.8±5.2 vs. 263.8±27.0 vs. 11.8±3.0 ng/24h, N=6, p<0.01). An intravenous infusion of sPRR-His counteracted the blood pressure-lowering effect of PF in Ang II-infused mice (MAP: 147±3 in PF+sPRR vs. 126±4 mmHg in PF; N=4, p<0.05). Together, these results suggest that S1P-derived sPRR contributes to Ang II-induced hypertension through activation of intrarenal renin-angiotensin system.
Abstract P238: A New Fluorometric Method to Evaluate Efficiencies of Carboxy-terminal Phenylalanine Cleavage From Angiotensin II, Angiotensin III and Apelin Peptides
Authors: Mueller Tilman , Wysocki Jan , Serfozo Peter D , Liu Pan , Ye Minghao , Jin Jing , Batlle Daniel
Published: July 2021

Angiotensin (Ang) II and apelins (AP) are intertwined by the actions of ACE2 that selectively cleaves the bond between proline and phenylalanine (Pro-Phe) at the carboxyl-termini of Ang II and AP leading to a conversion of Ang II (1-8) to Ang (1-7), and degradation of AP-13 and AP-36 peptides. Ang III (2-8) has been assumed - due to the presence of the Pro-Phe motif - but not yet experimentally demonstrated to be a substrate of ACE2. Measuring peptide substrates and products of these reactions are usually difficult using current ELISA assays due to their cross-reactivity. We sought to device a rapid and simple screening method that would provide readout of peptide products formation from reactions involving exclusively the c-terminal Phe cleavage. To this end we used a quantitative fluorometric assay recently described by us to compare the relative strength of mouse (m) and human (h) recombinant ACE2 in regards to cleavage of the c-terminal Phe from Ang II, Ang III, AP-13 and AP-36. Phe cleavage of AP-36 by mrACE2 was as efficient as its cleavage of AP-13 (2063±106 vs. 1670±163 RFU/ng rACE2, NS, n=5 experiments). mrACE2 formed twice as much Phe from both AP-36 and AP-13 than from Ang II. We further found that rACE2 also removes Phe from Ang III (2-8) to form Ang (2-7). This reaction for both mrACE2 and hrACE2 was as efficient as the cleavage of Phe from Ang II to form Ang (1-7) (for mrACE2 1096±136 vs. 683±56 and for hrACE2 1289±92 vs. 1479±69 RFU/ng protein, NS, respectively). The new fluorometric method detects cleavage of single c-terminal Phe from different peptides which are substrates of ACE2. A novel finding using this approach is the ability of rACE2 to cleave Phe from Ang III (2-8) to form Ang (2-7). This method will allow comparisons of efficiencies of recombinant enzymes in cleaving Phe from variety of bioactive peptides from their c-terminal Pro-Phe motifs and as such can constitute a useful tool for screening carboxypeptidase activities.
Abstract P240: Decarboxylation of Ang-(1-7) to Ala
1
-ang-(1-7) Leads to Major Changes in Pharmacodynamics
Authors: Tetzner Anja , Naughton Maura , Gebolys Kinga , Sala Esther , Villacañas Óscar , Walther Thomas
Published: July 2021

Background:
Within the renin-angiotensin system, angiotensin (Ang)-(1-7) is cardiovascular protective, stimulates regeneration, and opposes the often detrimental effects of Ang II. We identified two receptors for the heptapeptide; the G protein-coupled receptors Mas and MrgD. Recently, a decarboxylated form of Ang-(1-7), Ala
1
-Ang-(1-7) (Alamandine), has been described as having similar vasorelaxing effects as Ang-(1-7) but distinctively stimulating the MrgD receptor.

Methods and Results:
The aim of this study was to elucidate the consequences of the lack in the carboxyl group in amino acid one on intracellular signalling, to discover the receptor fingerprint for Ala
1
-Ang-(1-7), and to characterize the consequences for pharmacodynamics. Therfore cAMP was used as the main readout in receptor-transfected HEK293 cells as well as primary cells (mesangial cells and HUVEC). Ala
1
-Ang-(1-7) elevated cAMP concentrations in primary endothelial and mesangial cells. However, the dose-response curves clearly discriminated from the curves generated with Ang-(1-7) (EC
50
= 1.1 x 10
-8
M), with much lower EC
50
(EC
50
= 3.6 x 10
-11
M) and bell-shape for Ala
1
-Ang-(1-7). We provided pharmacological proof that both, Mas (EC
50
= 6.31 x 10
-12
M) and MrgD (EC
50
= 3.98 x 10
-13
M), are functional receptors for Ala
1
-Ang-(1-7). Consequently, the heptapeptide failed to increase cAMP concentration in primary mesangial cells with genetic deficiency in both receptors. As for Ang-(1-7), the AT2 blocker PD123319 also blocked the Ala
1
-Ang-(1-7) effects on Mas and MrgD receptors and in primary cells. The very distinct dose-response curves for both heptapeptides could be explained by
in silico
modelling, energy calculations, and an involvement of G
alpha i
for higher concentrations of Ala
1
-Ang-(1-7).

Conclusions:
Our results identify Ala
1
-Ang-(1-7) as a peptide with specific pharmacodynamic properties and build the basis for the design of more potent and efficient Ang-(1-7) analogues for therapeutic interventions in a rapidly growing number of diseases.
Abstract P245: Microrna-145-5p, a Potential Therapeutic Target of Regulating Vascular Calcification
Authors: Ahmed Abu S , Sheng Matilda H , Baylink David J
Published: July 2021

Vascular calcification (VC) occurs in most otherwise normal individuals over 60 years of age and is intimately linked to hypertension with severe cardiovascular complications, resulting in considerable morbidity and sometimes death. VC is characterized by the deposition of bone tissue within the vessel wall. The cause of VC is unknown although considerable progress has been made in characterizing its pathogenesis. There is direct or indirect evidence for several factors which could contribute to VC including, vitamin K deficiency, magnesium deficiency increased parathyroid hormone, increased serum phosphate, increased BMP activity and microRNAs (a small non-coding RNA molecules). The BMP pathway deserves attention because it is the sine qua non of osteoblast differentiation. With respect to microRNAs, these also could be relevant to the cause of VC because microRNAs are known to influence VC, and importantly, microRNAs can change with age. Here, we used an
in vitro
model to mimic VC using osteogenic medium in a vascular smooth muscle cell (VSMC) line. From microRNA analysis using quantitative real-time PCR we identified a potential key microRNA, miR-145-5p involved in vascular calcification. Specifically, we found that in our
in vitro
model of VC has a significant decrease in miR-145-5p expression. In silico analysis revealed that miR-145-5p may bind and regulate SMAD5, which mediates the action of BMP. Accordingly, we found an increase in SMAD5 expression at the mRNA level in VC condition
in vitro
. Based on the foregoing, we now propose the hypothesis that in young normal individuals, miR-145-5p normally suppresses SMAD5 activity in vasculature; whereas, in aging, miR-145-5p expression is impaired which would result in increasing SMAD5 gene expression, as well as its phosphorylation, which in turn would eventually lead to VC. Thus, miR-145-5p might be a potential therapeutic target to regulate VC in the elderly.
Abstract P246: Soluble (Pro)renin Receptor Induces Endothelial Cell Injury via Activation of Nox4/NF-κB Pathway
Authors: Fu Ziwei , Hu Jiajia , Wang Fei , Su Jiahui , Lu Xiaohan , Yang Tianxin
Published: July 2021

Circulating soluble (pro)renin receptor (sPRR), the cleavage product of the extracellular domain of PRR, is elevated in patients with cardiovascular diseases, highlighting its value as a disease biomarker. We explored a potential biological effect of sPRR in vascular endothelial cells. In primary human umbilical vein endothelial cells (HUVECs), a 2-h treatment with a recombinant histidine-tagged sPRR (sPRR-His) at 50 nM decreased IκBα protein by 55%, associated with 1.5-fold increase in nuclear fraction protein abundance of NF-κB p65 as assessed by immunoblotting. By immunostaining, the increased p65 was localized to the nucleus. The NF-κB p65 transcription factor activity assay on the isolated nuclear fractions showed that the activity increased 1.9-fold after 2-h sPRR-His treatment. At 3 h of sPRR-His treatment, mRNA expression was increased 3.8-fold for IL-6, 30.8-fold for IL-8, 17.3-fold for VCAM-1, and 4.2-fold for ICAM-1 as assessed by qRT-PCR; ELISA detected a 2.6-fold increase in medium IL-6 and 3.7-fold increase in medium IL-8. Moreover, enhanced apoptosis was evidenced by a 2.9-fold increase of cleaved caspase 3 protein and a 4-fold increase in the apoptotic cell number, as assessed by immunoblotting and annexin V-FITC/PI double staining flow cytometry, respectively. In addition, sPRR-His induced a 69% decrease in phospho-eNOS (Ser1177) protein abundance. At 2 h, sPRR-His induced a 2.0-fold increase in Nox4 protein and a 2.8-fold increase in medium H2O2 as assessed by immunoblotting and ROS-Glo H2O2 assay, respectively. A Nox1/4 inhibitor GKT137892 and NADPH oxidase inhibitor apocynin completely abolished sPRR-His-induced NF-κB activation, cytokine production, and apoptosis, similar to the effect of NF-κB inhibition with PDTC. Overall, the present study for the first time reports that sPRR serves as a pro-inflammatory and pro-apoptotic mediator in vascular endothelial cells through activation of Nox4/NF-κB. These findings support PRR/sPRR as a potential target for treatment of cardiovascular disease.
Abstract P249: Effects of Lifestyle Changes on Metabolism and Early Vascular Lesion in Healthy People
Authors: Fengcheng Xu , Chaoping Yu , Tianhu Liu
Published: July 2021

Objective:
Through propaganda and education on lifestyle change, we study the effects on metabolism and vascular lesions in healthy people.

Methods:
the healthy subjects that conform to the requirements, through propaganda and education on vascular health, through moderate exercise, proper control of starchy foods, low salt, low fat diet, reduce smoking and other lifestyle changes, compare changes in weight, renal function, fasting blood glucose, blood lipids and ankle brachial index (ABI), cardio ankle vascular index(CAVI) before and after lifestyle changes.

Results:
After lifestyle changed, the subjects’ body mass index [(23.13±3.18)kg/m
2
vs
(22.67±3.36)kg/m
2
], ABI[1.11±0.08
vs
1.09±0.09], CAVI[(7.14±1.13 )
vs
(7. 01±1.18) ], serum creatinine[(84.31±22.41)umol/L
vs
(79.92±23.64)umol/L], blood uric acid[(337.79±102.17 )umol/L
vs
(328.12±88.33)umol/L], low density lipoprotein cholesterol[(2.49±0.65) mmol/L
vs
(2.37±0.69) mmol/L],all have good changes.

Conclusion:
Healthy lifestyle is good for metabolism and early vascular lesions, can improve metabolic disorder and slow the occurrence of arteriosclerosis.
Abstract P284: Neuron Specific (Pro)renin Receptor Knockout Reduces Sodium Appetite and Attenuates the Development of DOCA-salt Induced Hypertension
Authors: Trebak Fatima , Li Wencheng , Feng Yumei
Published: July 2021

The (pro) renin receptor (PRR) is a key component of the renin-angiotensin system that is highly expressed in the brain. We previously showed that the neuronal PRR deletion attenuates deoxycorticosterone acetate (DOCA)-salt induced hypertension. However, the mechanism underlying remains unclear. To test our hypothesis that PRR is involved in the regulation of sodium appetite during DOCA-salt hypertension, we used a neuron-specific PRR knockout (PRRKO) mouse model generated using the Cre-LoxP system. The PRRKO and their wildtype controls (WT) were implanted with 50 mg DOCA pellet with free access to regular water and 0.9 % saline as the drinking solution. Blood pressure (BP) was monitored by telemetry system in conscious free moving mice. The fluid intake and urine output were monitored along 21 days of DOCA-Salt treatment. The BP is significantly lower in PRRKO compared with WT mice following 21 days of DOCA-salt treatment (112 ± 2 vs. 134 ± 7 mmHg, P= 0.0186). Interestingly, we found that saline intake (27.8 ± 1.8 vs. 15.9 ± 1.2 ml/day, P=0.0007) and total fluid intake (31.1 ± 1.9 vs. 21.1 ± 1.4 ml/day, P=0.003) were higher; while the regular water intake was lower (3.4 ± 0.6 vs. 5.2 ± 0.3 ml/day, P=0.03) in WT compared to PRRKO mice. PRR deletion in the neurons reduced sodium appetite presented as the ratio of saline intake over total fluid intake (0.75 ± 0.016 vs. 0.89 ± 0.019, P=0.0005), as well as total sodium intake (2.45 ± 0.19 vs. 4.28 ± 0.28 mmol/day, P=0.0007) compared with WT mice at the end of the protocol. In addition, the urinary sodium excretion was lower (13.3±1.17 vs. 20±1.17 mmol/day, P<0.0001), but not potassium excretion (0.64 ± 0.028 vs. 0.56 ± 0.05 mmol/day, P=0.1291) in PRRKO compared with WT mice; however, there is no difference in urine sodium and potassium concentrations. Furthermore, plasma vasopressin level (19.0 ± 2.7 vs. 33.6 ± 2.7 pg/ml, P=0.0037) is lower in the PRRKO compared with WT mice at the end of DOCA-salt treatment. In summary, PRR deletion in the neurons reduced sodium appetite, circulating vasopressin level, and attenuated the development of DOCA-salt induced hypertension. Taken together, the present findings suggest that PRR regulates the BP and plays a key role in salt-sensitive hypertension, at least in part, by modulating sodium appetite.
Abstract P291: Sodium-independent Dietary Effects on Renal Immune Cell Infiltration in Salt-sensitive Hypertension
Authors: Abais-Battad Justine M , Lund Hayley , Liang Mingyu , Liu Pengyuan , Mattson David L
Published: July 2021

Recent studies have shown sodium-independent dietary effects to be important in the development of Dahl SS hypertension and renal disease. Dahl SS/JrHsdMcwiCrl (SS/Crl) rats fed a grain-based diet (Teklad 5L2F) were less susceptible to salt-induced blood pressure elevation (116.5±1.2 vs 141.9±14.4 mmHg) and albuminuria (21.7±3.5 vs 162.9±22.3 mg/day) compared to SS/JrHsdMcwi (SS/Mcw) rats fed a 4.0% high salt casein-based diet (AIN-76A) for 3 weeks (n=6-8/group). Given the role of immunity in hypertension, SS/Crl rats displayed significantly less total CD45+ leukocyte infiltration in the kidney than SS/Mcw (6.1±0.9 vs 11.6±2.6 x10
6
cells/kidney). Reductions were observed in all subsequent immune cell populations, including CD3+ T cells (6.8±0.8 vs 14.4±2.2 x10
5
), CD45R+ B cells (1.5±0.4 vs 6.7±2.2 x10
5
), and CD11b/c+ antigen presenting cells (5.0±0.7 vs 9.2±2.1 x10
6
cells/kidney) in SS/Crl versus SS/Mcw. The SS/Crl immune system appears to be less activated, as demonstrated by a respective 44.8% and 66.8% reduction in CD4+CD25+ and CD8+CD25+ T cells, as well as a 40.4% decrease in TNFα-producing CD4+ T cells. Upon taking a discovery approach, RNA sequencing data (n=4 pools of 3 rats each) revealed genes related to hematopoietic cell lineage, the complement and coagulation cascade, B cell signaling, and primary immunodeficiency. Specifically, RNA expression of components of the NLRP3 inflammasome pathway were upregulated, including TLR4 (44.0%), NLRP3 (33.8%) and caspase-1 (26.4%), with a similar trend for IL-1β (25.4%), in the renal medulla of SS/Mcw rats fed high salt. This coincided with increased protein expression of NLRP3 (1.5-fold) and ASC (3.2-fold), together demonstrating increased NLRP3 inflammasome mRNA and protein expression in SS/Mcw rats after high salt challenge. Interestingly, RNA expression of these NLRP3 inflammasome proteins were significantly reduced in SS/Crl versus SS/Mcw rats fed high salt, including TLR4 (39.2%), NLRP3 (31.2%), caspase-1 (32.8%) and IL-1β (39.2%). These data indicate that the NLRP3 inflammasome may be a key mechanistic step in determining how these non-sodium components of the diet affect immunity, and warrant further studies to elucidate its role on salt-sensitive hypertension and renal damage.
Abstract P296: Deletion of Endothelial Sirt3 Causes Diastolic Dysfunction
Authors: He Xiaochen , Zeng Heng , Chen Jian Xiong X
Published: July 2021

The morbidity of heart failure with preserved ejection fraction (HFpEF) increases with advanced age and is associated with endothelial dysfunction. In the present study, we analyzed the cardiac function and coronary microvascular function in endothelial specific SIRT3 knockout (ECKO) mice and global SIRT3 KO mice, as well as endothelial glycolytic metabolism and angiogenesis
in vitro
. We found that SIRT3 ECKO mice developed left ventricle (LV) diastolic dysfunction, as evidenced by prolonged isovolumic relaxation time (IVRT) and reduction of coronary flow reserve. Western blot analysis revealed a significant decrease in glycolytic enzyme PFKFB3 in SIRT3 KO-ECs, accompanied by a dramatic reduction in basal glycolysis, glycolytic reserve and glycolytic capacity. Moreover, SIRT3 KO-ECs exhibited higher oxygen consumption rate and more prominent production of reactive oxygen species (ROS) than WT-ECs. SIRT3 KO-ECs exhibited less potent angiogenic capabilities as indicated by decreased network formation and migration. In contrast, global knockout of SIRT3 in mice resulted in progressive deterioration of LV function, as evidenced by a decrease in ejection fraction and fraction shortening at 12 months of age. SIRT3 KO mice also exhibited reduction of ANP and ERK-1/2, and increased ROS production, gp91(phox), caspase-3 and Wnt7 in the heart. In conclusion, we demonstrated that ablation of SIRT3 in ECs caused an HFpEF phenotype with impaired glucose metabolism and angiogenesis. Global SIRT3 KO mice, however, exhibited an HFrEF phenotype. Our data suggest cell-specific effects of SIRT3 deletion on cardiac function.
Abstract P347: γδ T Cells Drive CD4
+
And CD8
+
T Cell Activation In Hypertension
Authors: Caillon Antoine , Paradis Pierre , Schiffrin Ernesto L
Published: July 2021

Objective:
Both innate (monocyte/macrophages) and adaptive immune cells (T lymphocytes) have been shown to play a role in the development of vascular injury in hypertension. Recently, we demonstrated that a small subset of “innate-like” T lymphocytes, expressing the γ/δ T cell receptor (TCR) rather than the αβ TCR, plays a key role in hypertension and vascular injury. We demonstrated an increased number and activation (CD69
+
) of γδ T cells during the development of hypertension caused by angiotensin (Ang) II infusion, and that deficiency in γδ T cells prevented Ang II-induced hypertension, resistance artery endothelial dysfunction and spleen T-cell activation in mice. We hypothesized that γδ T cells mediate activation of other T cells in hypertension.

Method and Results:
Fourteen to 15-week old male C57BL/6 wild-type (WT) mice were infused with Ang II (490 ng/kg/min, SC) for 3, 7 and 14 days (n=5-7) and spleen T cell profile was determined by flow cytometry. A correlation was demonstrated between the frequency (FREQ) and the number (#) of activated CD69
+
γδ T cells and CD4
+
CD69
+
T cells (FREQ: r=0.41,
P
<0.05 and #: r=0.58,
P
<0.001) and CD8
+
CD69
+
T cells (FREQ: r=0.36,
P
<0.05 and #: r=0.50,
P
<0.01). We also demonstrated a high correlation between the # of CD69
+
γδ T cells expressing CD27, a marker of interferon-γ expressing cells and a member of the T-T interaction molecules, with CD4
+
CD69
+
(r=0.88,
P
<0.001) and CD8
+
CD69
+
(r=0.81,
P
<0.01) T cells after 7 days of Ang II infusion.

Conclusion:
This study demonstrated an association between CD27
+
CD69
+
γδ T cells and activated T cells. These results suggest that γδ T cells drive activation of other T cells in Ang II-induced hypertension. Targeting γδ T cells may contribute to reduce inflammation in hypertension.
Abstract P361: Prevalence & Predictors of Orthostatic Hypotension at a Tertiary Care Hypertension Clinic With New Diagnostic Thresholds
Authors: Hiremath Swapnil , Faraz Mohammad A , McCormick Brendan , Ruzicka Marcel
Published: July 2021

Background:
Orthostatic hypotension (OH), defined as a decrease of blood pressure (BP) of 20/10 mm Hg (systolic/diastolic) on change in posture from supine to standing is seldom assessed in routine practice because of logistical constraints. A recent study reported a sit-to-stand decrease of 15/7 mm Hg as also having good diagnostic yield. We measured the prevalence & risk factors associated with OH with the new threshold of sit-to- stand of either ≥ 15 mm Hg in systolic (SBP) or ≥ 7 mm Hg in diastolic BP (DBP).

Methods:
We reviewed medical charts of patients being followed at Renal Hypertension Center, a referral centre for difficult to control hypertension. Sitting BP is measured after 5 minutes of resting, as an average of 5 measurements with an automated device. Standing BP is measured three times at one minute intervals and averaged. OH was determined on the basis of the difference in either average SBP or DBP. Demographic characteristics, comorbidities, medication details, laboratory values and BP measurements were extracted.

Results:
Data from 219 patients was extracted (see table). The overall difference in SBP (sitting - standing)was 0.94 and DBP was 2.1 mm Hg. 190 patients (87%) did not have OH, whereas 29 (13%) had OH using either SBP or DBP thresholds. The difference in SBP and DBP was 17 mm and 6 mm Hg in those with OH, versus 1.6 and 3 mm Hg amongst those without OH respectively. Higher SBP was significantly associated with OH; age, gender, diabetes, number and hypertension drug class were not.

Conclusion:
Amongst referred patients to a specialist hypertension clinic, the prevalence of OH using a threshold of 15/7 mm Hg was 13%. The new diagnostic threshold allows for easy assessment of OH.
Abstract P365: Blood Pressure Evaluation From Ppg Signal Analysis and Artificial Neural Network
Authors: Lamonaca Francesco , Spagnuolo Vitaliano , De Prisco Serena , Carnì Domenico L , Grimaldi Domenico
Published: July 2021

The analysis of the PPG signal in the time domain for the evaluation of the blood pressure (BP) is proposed. Some features extracted from the PPG signal are used to train an Artificial Neural Network (ANN) to determine the function that fit the target systolic and diastolic BP. The data related to the PPG signals and BP used in the analysis are provided by the Multi-parameter Intelligent Monitoring in Intensive Care (MIMIC II) database. The pre-analysis of the signal to remove inconsistent data is also proposed. A set of 1750 valid pulse is considered. The 80% of the input samples is used for the training of the network. Instead, the 10% of the input data are used for the validation of the network and 10% for final test of this last. The results show as the error for both the systolic and diastolic BP evaluation is included in the range of ±3 mmHg. Tab.1 shows the results for 20 PPG pulses randomly selected analyzed together with the systolic and diastolic blood pressure furnished by MIMC and evaluated by the trained ANN.

Tab.1 experimental results comparing MIMIC and the ANN results.

Moreover, a suitable hardware to validate the ANN with the sphygmomanometer is designed and realized. This hardware allows clinicians to collect data according to the requirements of the validation procedure. With the sphygmomanometer the systolic and diastolic values are referred to two different PPG pulses. As a consequence, it is proposed a new hardware interface allowing the synchronized acquisition and storage of the PPG signal and clinician voice. For the validation, the clinician: (i) evaluates the BP on both the arms and assesses that no significant differences occur; (ii) plugs the PPG sensor on the finger of one arm; (iii) starts the recording of both the PPG signal and the audio signal; (iv) evaluates the BP on the other arm with sphygmomanometer and says the systolic and diastolic values when detected.

Through suitable post processing algorithm, the Systolic and Diastolic values are associated to the corresponding PPG Pulses. Following this procedure, the dataset to further validate the ANN according the standard is obtained.

Once the ANN is validated it will be implemented on smartphone to have always in the pocket a reliable measurement system for Blood Pressure, oximetry and heart rate.
Abstract P368: Effects of Empagliflozin on Day-to-day Variability of Home Blood Pressure and Heart Rate in Patients with Type 2 Diabetes
Authors: Misawa Tatsuo , Kawagoe Yumi , Oda Fujiko , Ogata Misao
Published: July 2021

Background:
The effects of empagliflozin in addition to standard care, on day-to-day variability of self-measured home blood pressure (BP) and heart rate (HR) at home in patients with type 2 diabetes (T2DM) at high cardiovascular risk are not known.

Method:
We followed twenty-three consecutive T2DM patients (mean age: 69 years old, 12 men) to add 10 mg of empagliflozin once daily for three months. Home BP and HR were measured once every morning at home, using an oscillometric device. The variability of BP and HR were defined as the standard deviations (SD) of measurements which were performed on seven consecutive days.

Results:
For home blood pressure, empagliflozin significantly reduced systolic blood pressure (SBP) from 130±11 mmHg at baseline to 126±11 mmHg at the first week(1W) of the administration (P<0.05). SBP achieved the target home BP goal (125±11 mmHg) at the second week(2W) and was maintained during the study (P<0.01). As regards day-to-day variability of SBP, SD decreased from 7.3±3.5 mmHg at baseline to 6.7±2.5 mmHg at 1W (4W: 6.4±2.8 mmHg, 8W: 5.2±2.3 mmHg, P<0.05). In diastolic blood pressure (DBP), there was a significant reduction of SD compared with that at baseline (4.9±1.6 mmHg at baseline, 4W: 4.3±1.6 mmHg, 8W: 3.9±1.8 mmHg, 12W: 4.4±1.6 mmHg, P<0.05); however, there was no change of DPB (71±10 mmHg at baseline, 12W: 71±8 mmHg). Similarly, there was a decreasing trend in SD of HR (3.9±1.0 beats per minute (bpm) at baseline, 4W: 3.3±1.3 bpm, 12W: 3.1±1.3 bpm, P<0.1), although there was no significant change in HR (62±13 bpm at baseline, 12W: 61 ± 12 bpm).

Conclusion:
Empagliflozin tended to reduce the day-to-day variability of self-measured morning home BP and HR.
Abstract P370: Nocturnal Blood Pressure Fall on Ambulatory Monitoring in a Cohort Data of Elderly Subjecsts
Authors: Lopes Patricia d , Rodrigues Livia F , Prado Yasmin S , Rocha Ligia d , Almeida Paulo E , Lucci Maria A , dos Santos Mariella P , Costa Lílian S
Published: July 2021

Introduction:
The use of Ambulatory Blood Pressure Monitoring has offered parameters that directly support stratification and independent association of cardiovascular risk (CV), especially in the elderly.

Objective:
To evaluate systolic (SBP) and diastolic (DBP) patient’s blood pressure (BP) on 24-hour, daytime and night time periods, pulse pressure (PP) and systolic and diastolic nocturnal drop in a population of elderly referred to be clinically healthy.

Method:
In a cohort of 1652 individuals over 60 years of age, both genders, we selected 434 subjects with no prior history of CV disease and off CV medications. Of these, 242 (56%) had normal and 192 (44%) had elevated mean 24-hour BP (G2), being 68 with isolated systolic hypertension (G2A) and 124 with systolic and diastolic hypertension (G2B). Analysis of variance was applied for a comparative study between groups, assuming a significance level of
p
less than 0.05.

Results:
Mean 24-hour, daytime and night time SBP in G1 were 118.94 ± 7.2, 122.27 ± 3.0, 110.23 ± 4.1 mmHg in men and 117.05 ± 6.3, 120, 46 ± 2.4 and 107.57 ± 3.4 mmHg in women (p = 0.06). Mean DBP values in the same periods were, respectively, 67.66 ± 6.62, 70.61 ± 5.3 and 60.24 ± 2.4 mmHg in men and 67.94 ± 5.2, 70.70 ± 5, 2 and 60.57 ± 5.1 mmHg in women (p = 0.08). Statistical analysis showed no difference for PP values between genders in different periods evaluated. In G2, there was a significant difference between subgroups A and B for all parameters, with mean 24-hour BP being 137,18x71,85 in G2A and 142,67x86,33 in G2B, p 0.003 SBP and 0.001 DBP, and 24-hour PP 65.32 and 56.33 mmHg, respectively (P 0.001), with no significant difference between genders. Regarding mean systolic and diastolic nocturnal drop, we observed values in G1 10,06 ± 7,07 and 14,37 ± 8,18, in G2A 5,24 ± 10,2 and 9,73 ± 9,8 and in G2B 8,93 ± 6.47 and 13.36 ± 6.5 mmHg, respectively; that is, lower values in G2A compared to G1 and G2B (p 0.001 and p 0.033), with G1 and G2B with no significant difference.

Conclusions:
Since the nocturnal drop pattern is the most consistent predictor of CV outcome, we have confirmed in our findings, previous literature data, which shown reduced nocturnal BP falls in the elderly population as a marker of BP elevation itself, after adjusting for age, gender and comorbidities.
Abstract P378: Current Progress of Antihypertensive Medication Use According to Healthy People 2020 Goals Among Adults With Hypertension- the National Health Interview Survey 2014-2015
Authors: Fang Jing , Ayala Carma , Loustalot Fleetwood
Published: July 2021

Background:
Hypertension is a major risk factor for heart disease and stroke; yet only half of those with hypertension have it under control. One of the Healthy People 2020 goals is to increase the proportion of adults with hypertension who are taking antihypertensive medications to lower their blood pressure from a baseline of 63.2% to a target of 69.5% by 2020. The objective of this study was to examine progress towards meeting this national objective and to assess disparities mid-decade.

Methods:
Adult participants (≥18 years) with self-reported hypertension from the National Health Interview Survey in 2014 and 2015 were included in this study. Current antihypertensive medication use was assessed and age-standardized for analysis. Multivariable logistic regression models were used to determine the odds ratio of antihypertensive medication use, adjusting for age, sex, race/ethnicity, education and health care access status.

Results:
Among the 21,050 (26.7%) adults with self-reported hypertension, 69.2% reported current antihypertensive medication use and disparities were noted among subgroups. Adults with estimates of antihypertensive medication use that fell below the Healthy People 2020 goal (69.5%) included women, non-Hispanic whites, Hispanics, and those with access health care barriers (Table).

Conclusion:
To control hypertension, most patients require antihypertensive medications to achieve control. Understanding disparities in antihypertensive medication utilization is needed for all population to achieve Healthy People 2020 targets by the end of the decade.
Abstract P403: Seasonal Variation in the Daily Urinary Sodium Excretion in Outpatients From the Northern Japan
Authors: Hashimoto Tomoko
Published: July 2021

Although the daily urinary sodium excretion (UNaV) is considered to provide the most reliable estimate of the daily sodium intake, it may be affected by salt loss due to sweating in summer. However, theseasonal variation in the daily UNaV associated with a normal lifestyle is unknown. This study was performed in 348 outpatients from the Morioka region during three seasons: summer(summer 1), winter, and the following summer (summer 2). The daily UNaV (g salt/day) was estimated by the second morning urine method three times during each season. Seasonal variation was defined as a significant trend across the three seasons together with a significant difference between winter and both summers. In women, the daily UNaV was higher in winter (11.8±3.0 g salt/day) than in summer 1 (11.2±2.9g salt/day) or summer 2 (11.0±2.9 g salt/day). In contrast, there was no marked seasonal variation in men. An analysis stratified by age (4 quartiles) identified seasonal variation in the older 2 quartiles of women (aged ≧68 years). In these women, the mean seasonal difference in the daily UNaV was 0.9 g of salt/day for both winter vs. summer 1 and winter vs. summer 2, while it was 0.1-0.8 g of salt/day in the other groups. Seasonal variation in the daily UNaV only occurred in older female patients and was relatively small. This is evidence for restricting salt intake throughout the year and should reassure patients who are anxious about salt loss due to sweating in summer.
Abstract P407: Five-year Longitudinal Bp In Treated Hypertensive Patients Shows Evidence For Diastolic J-curve And Cardiovascular Outcomes
Authors: Tan Li E , Lip Stefanie , McCallum Linsay , Touyz Rhian , Dominiczak Anna F , Padmanabhan Sandosh
Published: July 2021

Current guidelines recommend a BP target <140/90 mmHg irrespective of cardiovascular (CV) risk level. The SPRINT showed significant benefit of treating SBP to <120 mmHg. Concerns exist regarding the potential increased CV risk from intensive DBP lowering - the diastolic J-curve. We analysed the relationship of longitudinal on-treatment SBP and DBP on CV events.

Methods:
Longitudinal BPs were obtained from clinic records for hypertensive patients attending the Glasgow BP Clinic. AUC BP for the first year (Y1) and 2-5years (Y2-5) were calculated for patients who had >2 BP measurements during these periods. Survival analyses were performed using Cox proportional hazard model adjusted for age, sex, cholesterol, smoking status, BP (time-dependent) and Charlson comorbidity index (time-dependent).

Results:
There were 4813 eligible patients (mean age 54±14 years; female 52%). The average baseline, Y1 and Y2-5 BPs were 167/97±26/12, 153/91±19/9 and 146/87±16/8 mmHg respectively. There were 398, 452 and 455 first admissions with IHD in 15, 25, 35 years (31,563, 39,255 and 40,714 person-years respectively). DBP demonstrated a J-shaped relationship with IHD and MI events but not for CVA (Figure). Compared to DBP80-90 mmHg, DBP<80mmHg was associated with a nearly two-fold increased risk of IHD admissions 1.95[95%C.I. 1.46;2.61, P=7.6E-06]. Similar results were observed for 25- and 35-year follow-up periods.

Conclusions:
Low DBP (<90mmHg) is associated with increased risk of IHD but not CVA. Increasing SBP is linearly associated with increased risk of IHD and CVA. Therefore, intensive SBP reduction measures should recognise potential risks of concomitant DBP reduction.
Abstract P418: PPARγ Targets both (Pro)Renin Receptor and Site-1 Protease in the Collecting Duct to Mediate Rosiglitazone-induced Fluid Retention
Authors: Yang Tianxin , Wang Fei , Fu Ziwei , Zou Chang-Jiang , Peng Kexin , Hu Jiajia , Guan Guangju , Zhao Long
Published: July 2021

Collecting duct (CD)-specific deletion of PPARγ attenuates the fluid retention side effect of thiazolidinedione including rosiglitazone (Rosi) but the underlying mechanism largely remains unknown. (Pro)renin receptor (PRR) is a newly described regulator of CD function. Recent results have demonstrated site-1 protease (S1P) but not furin or ADAM19 as the predominant PRR cleavage protease. A histidine-tagged soluble PRR, sPRR-His, stimulates AQP2 expression. Here, we examined involvement of PRR/S1P during Rosi-induced fluid retention. PRR promoter contains two putative PRREs at positions -834~-828 bp (AGGTTA) and -318~-312 bp (GGTGCA). A 2.0-kb mouse PRR promoter showed a 10-fold increase in luciferase activity in mpkCCD cells exposed to 10 μM Rosi. The Rosi-induced response was first mapped to -421~ -219 bp by the deletion analysis and then to the second PPRE by mutagenesis. Rosi treatment at 80 mg/kg diet in C57/BL mice rapidly induced a 1.9-fold increase in renal PRR protein, a 2.4-fold increase in urinary sPRR, a 2.0-fold increase in urinary renin activity, a 48% reduction of urine volume, and a 4.8-fold increase in renal AQP2 protein, without an effect on expression of ENaC subunits, at 36 h, all of which were diminished thereafter. On day 3, Rosi induced a 9% body weight gain (BWG) and a 15% reduction of Hct, a 20% increase of fluorescein isothiocyanate dextran (FITC-d)-measured plasma volume. In contrast, the indices of Rosi-induced fluid retention along with AQP2 upregulation were all nearly abolished by a PRR antagonist PRO20 and CD PRR deletion. The sensitivity to Rosi-induced volume expansion as assessed by Hct and FITC-d in the null mice was fully restored by a 3-d i.v. infusion of sPRR-His at 30 μg/kg/d. A 2.1-kb mouse S1P promoter also contained two PPRE sites and showed a 3-fold increase in luciferase activity in response to Rosi. Administration of a S1P inhibitor PF-429242 via minipump in C57/BL6 mice attenuated Rosi-induced FITC-d-measured plasma volume (Control 1.28
+
0.1 vs. Rosi 1.61
+
0.05 vs. Rosi
+
PF429242 1.37
+
0.08 ml/kg), BWG, and Hct drop. Overall, PPARγ simultaneously targets PRR and S1P to generate sPRR to increase AQP2 expression to expand plasma volume during Rosi treatment.