bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P271: Interleukin-4 Supplementation Improves Chronic Inflammation and Hypertension in Response to Placental Ischemia During Pregnancy
Authors: Amaral Lorena M , Cottrell Jesse , Harmon Ashlyn , Cornelius Denise C , Cunningham Mark W , Wallukat Gerd , Herse Florian , Ibrahim Tarek , Dechend Ralf , LaMarca Babbette
Published: March 2022

Preeclampsia (PE), new onset hypertension in response to placental ischemia during pregnancy, is associated with chronic inflammation characterized by elevated tumor necrosis factor (TNF-α), interleukin-6 (IL-6), agonistic autoantibody to the Ang II type 1 receptor (AT1-AA). In addition, evidence for a shift towards proinflammatory CD4+TH1 vs CD4+TH2 profile exist in PE. Therefore we tested the hypothesis that IL-4, a cytokine essential for TH2 differentiation and proliferation, would improve pregnancy outcomes in response to placental ischemia (RUPP) by shifting the immune profile toward CD4+TH2. Interleukin-4, 600 ng/day, was administered via a mini-osmotic pump on day 14 of gestation into normal pregnant (NP) rats and into RUPP rats immediately following the reduced uterine perfusion pressure (RUPP) procedure and carotid catheters were inserted on gestation day 18. Blood pressure (MAP), TNF-α, IL-6, AT1-AA and circulating TH2 cells were measured on GD 19. MAP in NP rats (n=10) was 96
+
2, 101
+
2 in NP+IL-4 rats (n=14), 130
+
4 in RUPP rats (n=6), which improved to 108
+
2 mmHg in RUPP+IL-4 rats (n=15), p< 0.05. Plasma levels of TNF-α and IL-6 were 25
+
6, 29
+
3 in NP rats (n=7-8/group), 116
+
30, 224
+
62 in RUPP rats (n=9-10/group), which decreased to 23
+
5 and 135
+
48 pg/mL in RUPP+IL-4 rats (n=5-7/group), p<0.05. Importantly, plasma levels of AT1-AA were 0.3
+
0.4 in NP rats (n=8), 0.3
+
0.2 in NP+IL-4 rats (n=5), 18
+
0.3 in RUPP rats (n=9), which were blunted to 4
+
1 bpm in RUPP+IL-4 rats (n=14), p<0.05. Circulating TH2 cells were 17.3 ± 2.7 in NP rats (n=3), 9.0 ± 2.0 in RUPP rats (n=5), which improved to 13.4 ± 3.2 % gated in RUPP+IL-4 rats (n=9). This study illustrates that administration of IL-4 decreases TNF-α, IL-6, AT1-AA, possibly by increasing TH2/IL-4 in RUPP rats, which ultimately led to a reduction in hypertension in response to placental ischemia of pregnancy.
Abstract P290: Unilateral Cervical Vagotomy Protects Against Renal Injury and Hypertension in a Mouse Model of Lupus Hypertension
Authors: Pham Grace S , Chaudhari Sarika , Fancher Daniel L , Michaels Tanya B , D’Cunha Luke , Mathis Keisa W
Published: March 2022

Chronic renal inflammation has been implicated in hypertension. The vagus nerve controls inflammation by immunoregulatory mechanisms like the hypothalamic-pituitary-adrenal axis and the cholinergic anti-inflammatory pathway. It is not clear whether changes in vagal activity affect inflammation downstream in the kidney. We hypothesized that decreasing vagal neurotransmission would impede these anti-inflammatory mechanisms, exacerbate renal inflammation and promote hypertension. To study this, we used a mouse model of systemic lupus erythematosus (SLE) that develops hypertension after the onset of autoimmunity. Female SLE (
NZBWF1)
and control
(NZW)
mice received right unilateral cervical vagotomy (Vx) or sham surgery (n=4-6/group) and 3 weeks later had catheters implanted to measure blood pressure. Sham-operated SLE mice had elevated plasma double-stranded DNA autoantibodies (3.8 x 10
5
± 1.3 x 10
4
vs. 1.1 x 10
5
± 5.0 x 10
4
activity units; all p<0.05 unless otherwise noted), a hallmark of the disease; enlarged spleens (239.1 ± 80.4 vs. 124.1 ± 11.4 mg); and an elevated splenic CD3+CD4+ T cell population (flow cytometry; 21.9 ± 2.1 vs. 14.6 ± 2.8%) relative to controls. Vagotomy did not alter disease severity (3.2 x 10
5
± 3.5 x 10
4
; p = 0.20), but prevented splenomegaly (119.7 ± 24.2 mg) and reduced CD3+CD4+ T cells (14.7 ± 0.7) in SLE mice. SLE mice trended toward higher renal cortical TNF-α than controls (3.5 x 10
6
± 1.7 x 10
6
vs. 7.6 x 10
4
; normalized to total protein; p = 0.22), although SLE/Vx mice had lower concentrations than SLE/sham mice (4.9 x 10
5
± 1.3 x 10
5
vs. 3.5 x 10
6
± 1.7 x 10
6
). SLE mice had higher albumin excretion rates (AER) than controls (1.2 x 10
4
± 3.6 x 10
3
vs. 2.0 x 10
1
± 4.8 μg/day) and Vx decreased AER in SLE mice (1.2 x 10
4
± 3.6 x 10
3
). SLE mice were hypertensive compared to controls (158 ± 10 mmHg vs. 108 ± 4 mmHg) and vagotomy protected SLE mice from hypertension (133 ± 4). In sum, these data counter our initial hypothesis—unilateral vagotomy reduced renal inflammation and halted the progression of SLE hypertension. The vagus nerves have myriad functions and perhaps other neuroimmune interactions compensate for the ligation of one nerve. Future studies will investigate whether our findings may be due to long-term adaptation following unilateral vagotomy.
Abstract P299: Longitudinal Study of Angiotensins in Normal and Preeclamptic Pregnancy
Authors: Brosnihan K. Bridget , Merrill David C , Yamaleyeva Liliya C , Chen Kai , Joyner JaNee , Gruver Courtney , Lanier Kristen , Moorefield Carol
Published: March 2022

To address whether differential regulation of the renin-angiotensin system (RAS) occurs in preelampsia, we performed an extensive analysis of the time course of circulating and urinary profiles of the vasoconstrictor (Ang II) and the vasodilator [Ang-(1-7)] components in pregnant and preeclamptic women. Plasma and spot urine samples were collected prospectively from nulliparous normal pregnant (n=67) and preeclamptic (n=19) women between the ages of 18-40, at <16, 16-20, monthly between 24-36 weeks of gestation, and a control sample at six weeks post-delivery (CON). Mean blood pressure (p<0.001) was highest at 31-33 and 35-37 weeks of gestation in preeclamptic compared to normal pregnant women, and associated with mild proteinuria. Plasma Ang II was elevated in normal pregnant subjects at < 16 weeks of gestation and maintained at the higher levels over gestation (Fig A). In preeclamptic subjects plasma Ang II was elevated during gestation, but at 35-37 weeks plasma Ang II was reduced compared with normal pregnant subjects. Plasma Ang-(1-7) was unchanged in both normal and preeclamptic subjects (Fig B). Urinary Ang II was increased at 23-37 weeks of gestation in normal subjects and 27-33 in preeclamptic subjects (Fig C); urinary Ang-(1-7) was elevated in normal pregnant subjects (Fig D) but not in preeclamptic women. The activation of the RAS, particularly Ang II throughout normal gestation may contribute to the maintenance of vascular tone during normal pregnancy. However higher sensitivity to Ang II in preeclampsia may be potentiated by the higher circulating and urinary levels of Ang II, unopposed by local renal Ang-(1-7), and thus contribute to the development of preeclampsia.
Abstract P300: Fetal Status Evaluation Using Photoacoustics and 3D Power Doppler Ultrasound
Authors: Yamaleyeva Liliya M , Brosnihan K. B , Bledsoe Tiffaney
Published: March 2022

Pregnancy complications can seriously impact fetal well-being. However the tools used for the evaluation of fetal oxygenation in clinic are limited to the monitoring of fetal heart rate or invasive techniques (fetal scalp pH and PO
2
). Therefore, the development of methods enabling accurate, non-invasive and in real-time assessment of fetal status throughout pregnancy is warranted. In this study we tested the combination of photoacoustic imaging (PAI) and 3D power Doppler for the assessment of fetal distress. PAI combines optical contrast of photoacoustic laser technology with high spatial resolution of ultrasound. Power Doppler is a sensitive technique for the detection of blood flow. We hypothesize that a combination of PAI with 3D power Doppler can non-invasively and in real time establish fetal oxygenation, volume, and tissue vascularity. Pregnant C57Bl/6 mice were infused with nitric oxide synthase inhibitor, L-NAME via osmotic minipumps (50 mg/kg/day; days 11 to 14 of gestation) to induce the hypertensive phenotype. At day 14 of gestation, systolic blood pressures were higher in L-NAME-treated vs. untreated C57Bl/6 mice although these values did not reach hypertensive levels (90.8±3.1 vs. 104.4±3.7 mmHg, p<0.05, n=4-5). Fetal weights were lower in the L-NAME-infused mice versus controls (0.12±0.01 vs. 0.17±0.01 g fetal weight per cm tibia length, p<0.05, n=5-6). Fetal liver sO2 was lower in L-NAME-infused mice (47.5±2.1 vs. 56.1±1.6 %, n=4, p<0.05), while no differences were found in fetal brain sO2. Total fetal volume was lower in L-NAME-treated mice (213.8±26.5 vs. 399±45.1 %, n=5, p<0.05) and positively correlated with fetal body weights obtained postmortem in study groups. Fetal tissue vascularity was also lower in L-NAME-infused mice possibly due to limited vascular branching or blood flow (40.8±1.5 vs. 47.7±1.8%, p<0.05). These changes were associated with a compensatory increase in fetal heart rate in L-NAME-treated mice (170.7±9.1 vs. 135.8±5.7 %, p<0.05). We conclude that a combination of PAI with 3D power Doppler provides valuable information about fetal oxygenation and growth in association with tissue vascularity thus permitting non-invasive, in real-time analysis of fetal well-being.
Abstract P303: Endothelin Modulates Local Complement Activation in Placental Ischemia-induced Hypertension
Authors: Regal Jean F , Wing Cameron R , McCutcheon Luke , Gilbert Jeffrey S , Fleming Sherry D
Published: March 2022

Preeclampsia is characterized by hypertension, reduced placental perfusion, intrauterine growth restriction and increased activation of complement, part of innate immunity. Using the Reduced Uterine Perfusion Pressure (RUPP) model of placental ischemia-induced hypertension in rat, our previous work demonstrated the importance of complement activation in mediating maternal hypertension. Previous studies also demonstrated endothelin A receptor (ET
A
) plays a significant role in RUPP-induced hypertension, and RUPP-induced systemic complement activation occurs even with ET
A
antagonism with atrasentan (ATR). To further probe mechanistic connections between complement and endothelin, we hypothesized that ET
A
regulates local complement activation (C3 deposition). Dams received drinking water with 50 or 75 ug/ml ATR from gestation day (GD)13-19 corresponding to estimated average 5.5-6.6 and 8.0-9.3 mg/kg/day, respectively. On GD14 rats underwent Sham or RUPP surgery with clip placement on abdominal aorta and uterine arteries to decrease placental perfusion. As previously reported, RUPP increased mean arterial pressure (MAP) compared to Sham, and ATR attenuated RUPP-induced hypertension but not RUPP-induced placental C3 deposition (immunohistochemistry). Importantly, in Sham animals ATR increased C3 deposition and ATR 75 markedly decreased message for membrane bound placental complement regulators Crry, CD55 and CD59 that normally control complement activation. These data suggest that endothelin binding ET
A
modulates endogenous complement regulators to influence and prevent excess pathological complement activation in placenta during pregnancy.
Abstract P320: Cooperative Signaling and Membrane Recruitment of the Dopamine-1 (D
1
R) and Angiotensin Type 2 (AT
2
R) Receptors in Human Renal Proximal Tubule Cells (RPTC)
Authors: Gildea John J , Mallory Micah J , Felder Robin A
Published: March 2022

The D
1
R and AT
2
R are interdependent natriuretic receptors that are critical for the regulation of renal sodium balance and are implicated in both hypertension and salt-sensitivity. D
1
R and AT
2
R stimulation with the D
1
R agonist fenoldopam (FEN) and Angiotensin III (AngIII), selectively increases cAMP and cGMP, respectively, leading to D
1
R and AT
2
R recruitment to the cell surface (measured by fluorescent extracellular receptor epitope specific antibodies). The interdependence of the D
1
R and AT
2
R recruitment was investigated following AT
2
R stimulation with AngIII (10 nmol/L 1 hour) which leads to D
1
R cell surface recruitment using a normally D
1
R/Gs coupled cell line (i22) (VEH 8209±863, AngIII 19485±3425 RFU, n=6, p<0.05) but not in a D
1
R uncoupled from Gs cell line (i19). These data were verified by increasing intracellular sodium with monensin (100 μM, 1 hr) and measuring cell surface binding with a fluorescent labelled D
1
R agonist SKF83566 (10 nmol/L, 1 hr, i22 1.67±0.7 fold, n=5, p<0.0001, i19, NS). The importance of dopamine in regulating these receptor actions was shown by stimulating the AT
2
R with AngIII and measuring D
1
R recruitment followed by blockade of amino acid decarboxylase using either carbidopa or benzeraside (100 μM each, 1 hour). Additionally, FEN stimulated D
1
R surface recruitment is blocked by the AT
2
R inhibitor PD123319 (1 μM, 1 hour) and the coupling defect is fully rescued by 8Br-cGMP (1 mmol/L, 1 hour), a cell permeable second messenger normally thought to signal specifically through the AT
2
R. We then show that FEN stimulation leads to increased production of AngIII only in the D
1
R/Gs coupled i22 cell line (5.82±0.43 fold, n=6, p<0.05) but not in i19 the D
1
R/Gs uncoupled cell line and this production was blocked with an aminopeptidase A inhibitor, EC-33 (10 μmol/L, 1 hour). Addition of 8Br-cAMP (1mmol/L 1 hour) not only leads to an increase in AngIII production, but also cGMP production (4.33±0.36 fold, n=6, p<0.05) that is blocked by PD123319 (potent, selective, non-peptide angiotensin AT
2
R antagonist, 1 μM, 1 hour). In summary we not only show that dopaminergic stimulation leads to increased AngIII production in a D
1
R coupled manner, but also that AngIII stimulation of AT
2
R leads to dopaminergic activation in a D
1
R coupled manner.
Abstract P322: Interaction of Shroom3 With Fyn Impacts Phosphorylation of Nephrin Causing Proteinuria With Foot Process Effacement
Authors: Menon Madhav C , Philippe Nimrod , Wei Chengguo , Liu Ruijie , Zhang Weijia , Li Zhengzi , He John C , Murphy Barbara
Published: March 2022

In the translational GoCAR study, we identified that a CKD-associated SHROOM3-SNP, and tubular
Shroom3
expression correlated with the development of fibrosis in renal allografts. We showed that SHROOM3 facilitated TGF-B signaling suggesting its potential role as a therapeutic target. However, recent data suggest a
protective
role for SHROOM3 in proteinuria and glomerular development. To study the role of SHROOM3 in adult glomeruli, we used doxycycline-inducible (DOX), shRNA-mediated SHROOM3 knockdown, Podocin- and tubular-specific (PAX8)-RTTA mice, comparing these to non-transgenic DOX-fed littermates. After 2wks of DOX, adult Podo-RTTA mice developed significant albuminuria compared to littermates. Albuminuria was reversible on DOX-withdrawal, and reappeared on re-initiation. No podocyte loss [WT1 stain] was seen in these mice(8-wks DOX). EM revealed >50% foot process effacement. PAX8-RTTA mice did not show proteinuria. Glomerular RNA- seq identifed intracellular signaling/Small GTPAse signaling/integrin signaling/ actin-cytoskeleton among downregulated Gene-ontology terms in knockdown mice vs Controls. To identify protein interacting partners of SHROOM3, we performed mass spectrometry on protein lysates of 293-T cells overexpressing SHROOM3 immunoprecipitated (IP) with either anti-V5, -SHROOM3 or IgG. Among 491 unique interactions, we confirmed SHROOM3 as the top ranking protein. Interestingly, FYN - a src-kinase - was a top ranking candidate. Podocyte FYN is crucial for NPHS1-phosphorylation, and FYN-deficient mice show identical proteinuria phenotype. In human podocytes, we confirmed the interaction of endogenous SHROOM3 and FYN by IP. Glomerular protein extracts of Shroom3-knockdown mice showed decreased phosphorylation of FYN, and NPHS1. In human allografts from GoCAR, we identifed a corresponding reduced albuminuria (>1year post-transplant) associated with homozygosity of the risk allele in the donor. In summary, Podocyte-specific SHROOM3 knockdown causes a reversible proteinuria phenotype in adult mice, by interacting with FYN, a mechanism distinct from its effect on renal fibrosis in allografts.
Abstract P351: Cornell Product in Electrocardiogram is More Strongly Related to LV Regional Wall Motion Than Sokolow-Lyon Voltage
Authors: Ishikawa Joji , Yamanaka Yuko , Watanabe Shintaro , Harada Kazumasa
Published: March 2022

Backgrounds:
Left ventricular hypertrophy (LVH) evaluated by Cornell product in electrocardiogram predicted future stroke events greater than LVH by Sokolow-Lyon voltage. Therefore, we evaluated whether Cornell product was related to regional myocardial wall motion greater than Sokolow-Lyon voltage.

Methods:
We enrolled 288 hypertensive patients who were performed echocardiography for evaluating target organ damage. Cornell product was calculated as follows; R in aVL lead + S in V3 lead (0.6 mV added in female)хQRS duration; Sokolow-Lyon voltage, R in V5 lead + S in lead V1. We evaluated left ventricular mass index (LVMI), diastolic function of septal E/e’, and [longitudinal (GLS), radial (RS) and circumferential (CS)] strain using Altida (Toshiba, Japan).

Results:
Mean age was 63.4±13.2 years (male 47.9 %). There were 65.2 % of patients with antihypertensive medication. Both Cornell product, and Sokolow-Lyon voltage were related to LVMI (r=0.392, p<0.001 and r=0.315, p<0.001) and E/e’ (r=0.260, p<0.001; r=0.264, p<0.001), independently in multivariate linear regression analysis (both p<0.05). On the other hand, Cornell product was more strongly related, compared to Sokolow-Lyon voltage, to GLS (beta=0.200, P=0.005; beta=0.098, P=0.161), inner per outer ratio of RS (beta=-0.163, P=0.031; beta=0.117, P=0.116), and inner per outer ratio of CS (beta=0.148, P=0.044; beta=0.078, P=0.279).

Conclusions:
Both Cornell product and Sokolow-Lyon voltage were independently related to LVMI and diastolic function measured by tissue Doppler image; however, Cornell product was more strongly related to left ventricular regional wall motion of global longitudinal strain and trasmurality of radial and circumferential strain, than Sokolow-Lyon voltage.
Abstract P363: Dual Monitor Protocol Comparing the Accuracy of the Oscar 2 & Spacelabs 90207 24-hr Ambulatory Blood Pressure Monitors
Authors: Lombardi V P , Reichhold P C , Dietz N R , Pate D L
Published: March 2022

Background:
Our dual monitor protocol enables simultaneous blood pressures (BPs) using two ambulatory monitors (ABPMs) and two observers with a mercury (Hg) column. In normotensives, hypertensives and alcoholics, an
auscultatory
ABPM underestimated diastolic BP (DBP) progressively from supine to seated to standing. Our goal was to test the accuracy and reliability of
oscillometric
ABPMs.

Methods:
We programmed an Oscar 2 and Spacelabs 90207 to record simultaneous, opposite arm measurements for 24 hr in a hypertensive male (62 yr). Clocks were synchronized and cuffs were switched every 2-3 hr during waking hr. In the lab observers assessed simultaneous same arm BPs using an Hg column and ABPMs recorded simultaneous opposite arm BPs. Lab techniques were repeated in two young (21 yr) male normotensives.

Results:
The monitors’ SBP, DPB and mean arterial pressure differed significantly (all p < 0.001) for 109 simultaneous 24-hr measures. The Oscar averaged 154/101 ± 12/9 mm Hg (Stage II), while the Spacelabs 146/98 ± 10/8 (Stage I), differing by 8/3 ± 7/9 mmHg. Both machines exhibited troubling run-away inflations during walking and stair climbing as sustained motion artifacts were interpreted as not achieving MAP or peak SBP. The ABPMs common variance for SBP was 63% (R = 0.79) and for DBP was 48% (R = 0.69), significantly lower than BPs for trained clinicians using an Hg column (SBP 97%, DBP 95%). In the lab, for SBP the Oscar (151 ± 3 mm Hg) was significantly (P<0.05) higher than the Spacelabs (143 ± 3 mm Hg) and observers (144 ± 4 mm Hg). For DBP, the Spacelabs (88 ± 3 mm Hg) tended (P=0.051) to be lower than the Oscar (90 ± 3 mm Hg) and observers (92 ± 4 mm Hg).

Conclusions:
Subjects can be classified differently based on the ABPM model used. The Oscar and Spacelabs ABPMs demonstrate greater variability estimating DBP. If results from multiple ABPMs are pooled in meta-analyses, interpretations may be muddled especially if one is overestimating and another underestimating 24-hr pressures. Though our dual monitor protocol is challenging to implement, we hope to conduct studies in a large, diverse group. US and international protocols should require postural and exercise testing to ensure that devices truly can be classified as ambulatory and are more consistently accurate and reliable.
Abstract P372: Validation of a m-Health Platform for Blood Pressure Control and Reduction
Authors: Peretto Giovanni , Del Mauro Gaia , Bordignon Tommaso , Colombo Eros , Fabi Martina , Negri Stefano , Vergani Vittoria , Cianflone Domenico
Published: March 2022

Background:
Self-measured blood pressure (BP) monitoring, has been proposed as a means of improving BP control. Lifestyle modifications are indicated in hypertension at any stage.
AHA’s Life Simple 7
provides a straightforward call-to-action. mHealth services (mHs) may provide additional tools to effective BP control.

Aim:
We tested the reproducibility of the initial results of a mHs for remote BP monitoring and lifestyle counseling. The mHs in study is a
cloud
-based BP -recording, -interpretation and -trend evaluation service (BT) coupled with personalized lifestyle-change programs (LC). Initial results showed a significant reduction of BP at the end of the LC program. BT collects BPs, and after consistency-checks, feeds a proprietary CE-certified medical device algorithm to provide actual BP interpretation and BP trend assessment. Through a separate proprietary algorithm, LC provides a 3-month personalized lifestyle modification program for BP reduction, built on the
AHA’s Life Simple 7
, after collecting detailed clinical and lifestyle data. BP changes were evaluated comparing BPs recorded at the beginning and at 3 months in both groups. We studied 2601 subjects (50±14 years old) that voluntarily and unsolicited joined LC+BT or BT-only, from June 2016 to April 2017, via App or website.

Results:
During the study period a total 60044 BP values were collected, subjects with discontinuous BP measurements were excluded from evaluation. We hence analyzed the data of 575 LC+BT subjects and 1653 BT-only subjects. We observed a significant BP reduction in LC+BT subjects at the end of the program for both systolic and diastolic BPs: 4.7±1.2 and 3.3±0.8 (mean
+
SEM, mmHg), p<0.0001, respectively. In BT-only subjects the reduction of BP values at 3-months was
2.5 ±0.9 and 1.7±1.0, p<0.001, respectively.
The subjects of the LC+BT group with
>
3 systolic BPs above normal values in the first 10 days, experienced a greater BP reduction:
7.9±1.4 and 4.2±1.1, p<0.0001, respectively.

Conclusions:
The data confirm that this mHealth service provides a strong and effective adjunctive tool for BP reduction. These results support the value of the combination of algorithm based remote BP monitoring and a personalized implementation of
AHA’s Life Simple 7
recommendations.
Abstract P392: Autonomic Reflexes Mask the Hemodynamic Effects of Continuous Positive Airway Pressure
Authors: Okamoto Luis E , Gamboa Alfredo , Celedonio Jorge E , Diedrich Andre , Paranjape Sachin Y , Biaggioni Italo
Published: March 2022

Continuous positive airway pressure (CPAP) improves cardiac function and blood pressure (BP) in patients with sleep-breathing disorders by reducing increased sympathetic tone. The direct hemodynamic effects of increased intrathoracic pressure with CPAP, however, are not known and are likely masked by baroreflex buffering. To determine the acute hemodynamic effects of CPAP in the absence of autonomic modulation, we applied 5 levels of CPAP (0, 4, 8, 12 and 16 cm H
2
O, each for 2 min) to 7 healthy normotensive (NTN, 33±3 years, BMI 29±3, 4 females) and 12 hypertensive subjects (HTN, 50±2 years, BMI 30±2, 8 females) before and during autonomic withdrawal with the ganglionic blocker trimethaphan. Hemodynamic parameters were measured at the end of each CPAP level. At baseline, CPAP had no effect on systolic BP (SBP), heart rate (HR), cardiac output (CO), stroke volume (SV) or systemic vascular resistance (SVR) in either group. During the autonomic blockade, CPAP significantly decreased SBP in both groups (Figure). The maximal BP drop was twice as much in HTN than NTN (-17±2 vs. -8±2 mmHg at 16 cm H
2
O; P<0.02) but the final SBP was the same in both groups (85±3 mmHg). In HTN, CO and SV decreased -25±4% and -24±4% at 16 cm H
2
O, respectively; whereas in NTN, the decreases were -20±5% and -24±4%. Neither SVR nor HR changed significantly with CPAP in either group. We conclude that, in the absence of autonomic modulation, CPAP acutely decreases SBP significantly, due to decreases in CO and SV likely reflecting a decrease in venous return and increase in venous capacitance. The effect on BP was greater in HTN suggesting that a sympathetically mediated reduction in capacitance may contribute to the hypertension.
Abstract P400: Relative Contributions of Systolic and Diastolic Blood Pressure to Adverse Cardiovascular Outcomes in an Outpatient Cohort of 1.3 Million Adults
Authors: Flint Alexander C , Conell Carol , Ren Xiushui , Banki Nader , Chan Sheila L , Rao Vivek A , Melles Ron B , Bhatt Deepak L
Published: March 2022

Systolic hypertension is believed to have a greater influence than diastolic hypertension on adverse cardiovascular events. We tested this hypothesis using data from an integrated healthcare system, with 36.8 million measures from 1.3 million adults. Increasing SBP was associated with risk of a composite outcome of MI, ischemic stroke, or hemorrhagic stroke (Fig A). DBP showed a very different relationship with outcomes, with higher risk at the lowest and highest levels of DBP (Fig B). We examined the impact of systolic and diastolic pressures in multivariable survival analysis controlling for age, sex, race, and comorbidities using two different approaches. We first used the burden of systolic (SBP>=140) and diastolic (DBP>=90) hypertension as predictors. Both systolic burden (hazard ratio [HR] 1.17 for z-score, 95% CI 1.16 to 1.18, P<0.001) and diastolic burden (HR 1.07 for z-score, 95% CI 1.06 to 1.08, P<0.001) were associated with outcomes (Fig C). To examine a broader range of pressures from normal to elevated, where both measures are directly correlated with outcome, we used weighted average SBP and DBP from subjects above the 75th percentile for SBP (>133) or DBP (>78). Again, both SBP (HR 1.39 for z-score, 95% CI 1.37 to 1.41, P<0.001) and DBP (HR 1.24 for z-score, 95% CI 1.22 to 1.26, P<0.001) independently predicted outcomes (Fig D). A complicated relationship exists between SBP, DBP, and outcomes; at mid-normal to elevated pressures, both SBP and DBP are independently associated with the risk of adverse cardiovascular events.
Abstract P433: Impact of Renal Function on Outcomes After Percutaneous Transluminal Renal Angioplasty in Hypertensive Patients With Renal Artery Stenosis
Authors: Iwashima Yoshio , Fukuda Tetsuya , Horio Takeshi , Hayashi Shin-ichiro , Kusunoki Hiroshi , Kishida Masatsugu , Kamide Kei , Kawano Yuhei , Yoshihara Fumiki
Published: March 2022

Objective:
Atherosclerotic renal artery stenosis (ARAS) is associated with secondary hypertension. Renal dysfunction is often present in ARAS patients; however, evidence for the impact of pretreatment renal function on outcomes after percutaneous transluminal renal angioplasty (PTA) is limited.

Methods:
A total of 139 hypertensive patients with ARAS (mean age, 69.5±8.9 years, 112 male) who underwent renal PTA were included. Renal function was evaluated based on estimated glomerular filtration rate (eGFR) and albuminuria/proteinuria, and classified into three categories according to eGFR (≥45, 30-44, <30 ml/min/1.73m
2
) and albuminuria/proteinuria [normal-to-mild: albumin/creatinine ratio (ACR) <3.0, protein/creatinine ratio (PCR) <15; moderate: ACR 3.0-30.0, PCR 15-50; severe: ACR >30.0, PCR >50 mg/mmol]. The primary end point of this study was first occurrence of the composite of cardiovascular and renal events including all-cause death, myocardial infarction, stroke, adverse aortic events, or end-stage renal failure requiring regular hemodialysis.

Results:
During a median follow up of 5.4 years, 36.0% (50 of 139) of patients developed the primary composite end point including cardiovascular and renal outcomes. In multivariate Cox regression analysis, eGFR <30 (hazard ratio [HR] 3.47, p <0.01) as well as severe albuminuria/proteinuria (HR 2.63, p <0.05) was an independent predictor of worse outcome. In the subgroup without events within one year after PTA (n=117), the outcome differed among the three renal functional categories at one year based on eGFR (log-rank χ
2
=16.28, p <0.001) as well as on albuminuria/proteinuria (log-rank χ
2
=8.30, p <0.05). At one year, 24 of 117 patients (20.1%) showed a ≥20% decrease in eGFR, and their outcome was worse than that of those with a ≥20% increase (n=23) (HR 3.50, p <0.05). Multiple logistic regression analysis indicated that pretreatment moderate-to-severe albuminuria/proteinuria was an independent predictor of a ≥20% eGFR decrease (odds ratio 2.82, p <0.05).

Conclusion:
Impaired renal function, and in particular, a poor response of eGFR to PTA, is associated with worse outcome. Therapeutic effectiveness of renal angioplasty seems to be limited in patients with albuminuria/proteinuria.
Abstract P453: Hypertension Mortality Risk May be Fake News for Nonagenarians
Authors: Kheirbek Raya E , Alemi Farrokh , Fletcher Ross D
Published: March 2022

Background:
Little is known regarding BP control and mortality risk in subjects greater than 90 years of age.

Objective:
This paper assesses the association between systolic blood pressure and mortality risk for nonagenarians.

Methods:
Data from the Veterans Administration Informatics and Computing Infrastructure were used to analyze the survival of 193,651 nonagenarians in 130 Veteran Administration medical centers. Following clinical guidelines, for each day, we selected the lowest Systolic reading of the day exceeding 90 mmHg and defined sustained pressure as the average of two consecutive readings at least one month apart. Kaplan Meier curves and Cox regression was used to analyze survival.

Results:
Odds of mortality from hypertension declined with age. When patients were over 90 years old, the odds of mortality for hypertensives was below 1 to 1, suggesting a protective effect. Patients whose sustained systolic pressures exceeded 140 mmHg survived longer than patients whose highest sustained pressure was between 90 mmHg to 140 mmHg (p<.0001). Furthermore, strict control of hypertension of 90 year old patients (SBP 120) was associated with lower days of survival than hypertensives, raising questions about value of strict control of hypertension among 90 year olds.

Conclusions:
For nonagenarians, mortality from hypertension may be a lower concern than mortality from other causes. Randomized clinical trials are needed to examine the impact of control of hypertension of patient over 90 year olds.
Abstract P459: The BBSome Mediate the Sorting of the Serotonin 5-HT2C Receptor to the Plasma Membrane in POMC Neurons
Authors: Guo Deng-Fu , Searby Charles , Zhang QiHong , Nishimura Darryl , Sheffield Val , Rahmouni Kamal
Published: March 2022

The BBSome, a multiplex of 8 Bardet-Biedl Syndrome (BBS) proteins including BBS1, has emerged as an important regulator of energy homeostasis and cardiovascular function. Disrupting the BBSome, through deletion of its units, globally, in the nervous system, hypothalamus or selective neurons such as the proopiomelanocortin (POMC) neurons causes obesity associated with sympathetic overdrive and hypertension. However, the mechanisms underlying these phenotypes evoked by BBSome disruption remain ill defined. Serotonin 5-HT2C receptor (5-HT2CR) present in POMC neurons plays an important role in the regulation of energy homeostasis. We hypothesized that the BBSome contribute to the regulation of 5-HT2CR signaling. Given the importance of the BSBome for the trafficking of proteins to cilia, we first assessed whether 5-HT2CR localize the cilium in mouse MICD cells transfected with GFP-tagged 5-HT2CR. We found 5-HT2CR present in the plasma membrane, but no evidence for ciliary localization of this receptor. To study the relevance of the BBSome for the trafficking of 5-HT2CR, we deleted the
Bbs1
gene in IMCD cells using CRISP-Cas9. We found that the 5-HT2CR failed to traffic to the plasma membrane in IMCD cells lacking the
Bbs1
gene. Moreover, the increase in intracellular calcium and membrane potential evoked by serotonin were significantly reduced in cells lacking the
Bbs1
gene. Interestingly, double immunostaining revealed that most of 5-HT2CR was stuck in the late endosome co-localizing with CD63, a marker of this organelle. Next, we analyzed the localization of the 5-HT2CR in the hypothalamic arcuate nucleus of mice lacking the
Bbs1
gene selectively in POMC neurons (Bbs1
POMC
). In control mice, 5-HT2CR was predominantly present in the plasma membrane of POMC neurons and in some non-POMC cells. In Bbs1
POMC
mice, 5-HT2CR was altered selectively in POMC neurons appearing stalled in the cytoplasm. Finally, i.p. administration of lorcaserin, a 5-HT2C specific agonist, caused a significant decrease in body weight and food intake in control mice, but not in Bbs1
POMC
mice. These findings demonstrate that the BBSome underlie the trafficking of 5-HT2CR in POMC neurons with important implications for the regulation of energy homeostasis and development of obesity.
Abstract P503: Role of TRPV4 and Endothelial BK Channels in Endothelial-dependent Dilation Following Chronic Hypoxia
Authors: Naik Jay S , Walker Benjimen R
Published: March 2022

Following chronic hypoxia (CH) the systemic vasculature exhibits blunted vasoconstriction due to endothelial cell-dependent hyperpolarization (EDH). Previous data demonstrate that after CH, EDH-mediated dilation switches from a reliance on SK
ca
and IK
ca
channels to activation of endothelial BK
ca
channel (eBK). The mechanism by which endothelial cell stimulation activates eBK channels following CH is not known. We hypothesized that following CH, EDH-dependent dilation involves a transient receptor potential V4 (TRPV4)-dependent activation of eBK channels. ACh induced concentration-dependent dilation in pressurized gracilis arteries from normoxic and CH rats. TRPV4 blockade only attenuated ACh-induced vasodilation in arteries from CH rats (Fig. 1). ACh elicited an increase in endothelial TRPV4-dependent Ca
2+
events in arteries from both groups. Direct activation of TRPV4 elicited dilation in arteries from normoxic and CH rats (97 ± 2 % and 95 ± 2 % for normoxic and CH, respectively). In arteries from normoxic rats, inhibition of SK
ca
and IK
ca
abolished dilation to TRPV4 activation (6 ± 5 %), whereas, luminal iberiotoxin had no effect (89 ± 7 %). In CH rats, only administration of all three K
ca
channel inhibitors abolished dilation to TRPV4 activation (3 ± 10%). Disruption of endothelial caveolae with methyl-β-cyclodextrin significantly decreased ACh-induced dilation in arteries from both groups. Using a proximity ligation assay, we observed co-localization between caveolin-1, TRPV4, and eBK channels in the endothelium of gracilis arteries. In conclusion, CH results in functional coupling between muscarinic receptors, TRPV4 and K
ca
channels in gracilis arteries.
Abstract P505: Longitudinal Associations Among Flow-mediated Vasodilatation, Preclinical Vascular/renal Damage and Serum Cholesterol Levels in Treated Hypertension
Authors: Tomiyama Hirofumi , Yamashina Akira , Group FMDJ Study
Published: March 2022

Objective:
The endothelial dysfunction is thought to be an early precursor of vascular/renal damages. The present study was conducted to examine whether endothelial dysfunction, as assessed by measurement of flow-mediated vasodilatation (FMD), is longitudinally associated with the progression of vascular/renal damage, and also to identify factors that can perpetuate endothelial dysfunction in treated hypertension.

Methods and Results:
A 3-year multicenter prospective observational study was conducted in 674 Japanese patients with hypertension receiving antihypertensive medication, in whom the carotid intima-media thickness/plaque (CIMT), brachial-ankle pulse wave velocity (baPWV), estimated glomerular filtration rate (eGFR), urinary albumin/creatinine excretion ratio (UACR) and FMD were measured thrice at 1.5-year intervals. A lower value of the FMD at the study baseline was associated with a higher rate of increase of the baPWV during the study period (The change of baPWV during the study period was -25 cm/sec in subjects with the highest quartile range of FMD vs. +18 cm/sec in subjects with the lowest quartile range of FMD, p<0.05). However, no association of the FMD at the study baseline was observed with any of the other markers of vascular/renal damage examined. Mixed model linear regression analysis revealed a significant inverse relationship of the FMD at each measurement point with the baPWV at each measurement point (beta estimate = -10.97, P < 0.01); furthermore, the serum level of low density lipoprotein cholesterol (LDLC) at the study baseline was also inversely associated with the longitudinal changes of the FMD values (estimate = -0.39, P < 0.01).

Conclusion:
Our results suggest that an “endothelial dysfunction-arterial stiffening continuum” may exist in treated hypertension, and endothelial dysfunction, as measured by FMD, may be more closely associated with the arterial stiffness than with other parameters of preclinical renal/vascular damage. Furthermore, inadequate control of LDLC may also contribute to the progression of endothelial dysfunction.
Abstract P510: Nox4 Deficiency Leads to Hypertension and Vascular Damage With Enhanced Effects in Ang II-dependent Hypertension
Authors: Lacchini Silvia , Montezano Augusto C , Lopes Rheure A , Harvey Adam P , Rios Francisco J , Schroder Katrin , Brandes Ralf P , Touyz Rhian M
Published: March 2022

We previously showed that Nox1 and Nox2 are not involved in chronic Ang II-dependent hypertension, which recapitulates human hypertension. Here we questioned the role of Nox4 by studying transgenic mice expressing human renin (LinA3) crossed with Nox4-/- mice. Four groups were used: wildtype (WT), LinA3, Nox4 KO (Nox4), and LinA3/Nox4 KO (LinA3/Nox4). Blood pressure was measured by tail cuff. Aorta was collected to assess wall thickness, collagen and glycosaminoglycans (GAGs) deposition, and TNFα expression. Mesenteric arteries were used to access vascular function by myography. Blood pressure was increased in LinA3, Nox4 and LinA3/Nox4 mice vs WT (p<0.05). All three experimental groups exhibited vascular remodeling with evidence of increased fibrosis. Although LinA3 had increased aortic wall thickness (+31%), there was no significant change in collagen (10.3±3 vs. 8.5±2% in WT) and GAGs (6.6±3 vs 2.8±2% in WT) deposition (p<0.05). Nox4 mice, which presented a similar increase in wall thickness to LinA3 (+31%), had significant increase in collagen (20.6±6%) and GAGs (22.3±4%) in aorta (p<0.05). In LinA3/Nox4 mice, collagen (24.6±7%) and GAGs (37.1±10%) deposition were increased vs LinA3. TNFα was increased in LinA3 (130.4±6 a.u.) and LinA3/Nox4 mice (129±5 a.u.) vs WT (116.8±9 a.u.) (p<0.05). Mesenteric arteries from LinA3, Nox4 and LinA3/Nox4 mice, exhibit increased Phenylephrine-induced vasoconstriction vs WT (Emax: WT 6.79±0.29 vs LinA3 9.37±0.51; Nox4 9.87±1.59; LinA3/Nox4 9.12±1.63, p<0.05). Endothelium-dependent vasodilation was not reduced in Nox4 but impaired in LinA3 and LinA3/Nox4 (Emax: WT 86.48±0.01 vs LinA3 59.70±0.03; LinA3/Nox4 33.57±0.26, p<0.05). In conclusion, Nox4 deficiency was associated with increased blood pressure, vascular dysfunction and fibrosis, effects that were variably enhanced in LinA3/Nox4 mice. We also observed that the fibrosis in vessels from Nox4 mice was not associated with inflammation. These results suggest that Nox4 may be cardiovascular protective, which when downregulated leads to blood pressure elevation and vascular injury, processes that may be amplified by Ang II-dependent hypertension.
Abstract P526: MMP-2 Activation By S-glutathiolation Decreases Calponin-1 During Vascular Remodeling In Hypertension
Authors: Parente Juliana , Mello Marcela , Chan Brandon , Schulz Richard , Castro Michele
Published: March 2022

Aim:
Antioxidant tempol may decrease MMP-2 activation by S-glutathiolation as it reduces oxidative stress, and then prevents calponin-1 cleavage and aortic remodeling in hypertension.

Methods:
Male Wistar rats were submitted to the two kidney-one clip (2K-1C) model of hypertension or sham surgery and were treated with tempol (18 mg/kg/day) or water by gavage for one week. Systolic blood pressure (SBP) was accessed by tail-cuff plethysmography. Aortas were used to perform
in situ
zymography, immunofluorescence for dihydroethidium and calponin-1, immunohistochemistry for nitrotyrosine, co-immunoprecipitation for glutathione followed by Western blot for MMP-2 and histomorphology. Two-way ANOVA followed by Bonferroni test was done (Ethics Committee approval number: 0015/2017).

Results:
SBP increased in 2K-1C rats compared to Sham group (156±2.9 vs. 128.6±4.1, p<0.05) and tempol did not reduce it (147.3±4.7, p>0.05). Morphological analysis showed increased that 2K-1C aortas had increased media to lumen ratio (M/L) (8.6±0.7 vs. 6.5±0.2) and cross-sectional area (CSA) (1.4x10
6
±1.5x10
5
vs. 9.7x10
5
±4.2x10
4
) (p<0.05 vs. Sham), but tempol did not decrease them (M/L: 7.9±0.8; CSA: 1.3x10
6
±1.3x10
5
, p>0.05 vs. 2K-1C). However, tempol decreased 2K-1C-induced increased nitrotyrosine levels (5.9x10
5
±1.3x10
4
vs. 4.0x10
5
±6.1x10
4
), dihydroethidium (16.9±1.7 vs. 8.9±1.1) and MMP-2 activity (57.5±6.4 vs. 39.4±2.6; p<0.05). All four groups presented S-glutathiolation of MMP-2, however, the levels of calponin-1 were only reduced in aortas from 2K-1C rats (17.2±1.5 vs. 28.5±1.2, p<0.05 vs. Sham groups) and tempol reverted such loss (23.6±1.6, p<0.05 vs. 2K-1C). As calponin-1 is a potential intracellular target of MMP-2 in VSMC, these results suggest that the intracellular activation of MMP-2 may be more pronounced in hypertension. Different concentrations of tempol (0.5, 1 or 1.5 mM) did not

directly inhibit MMP-2 activity in gelatin zymography (respectively: 4.6x10
6
±7.4x10
4
, 4.9x10
6
±1.2x10
5
and 4.8x10
6
±1.5x10
5
vs. 4.6x10
6
±2.2x10
5
; p>0.05).

Conclusion:
Increased oxidative stress may contribute to activate aortic MMP-2 by S-glutathiolation in early hypertension, thus resulting in calponin-1 loss and maladaptive remodeling.
Abstract P543: Changes in Plasma Anandamide Concentrations are Associated With Reduction in Blood Pressure in Morbidly Obese Patients Undergoing Bariatric Surgery
Authors: Oliveras Anna , Pastor Antoni , Granados Ana Marina , Goday Albert , Ramon-Morós José M , Vázquez Susana , Vera Maria , Sans Laia , de la Torre Rafael , Pascual Julio
Published: March 2022

The role of endocannabinoids in the vascular system is not well-known. Blood pressure (BP) levels decrease in obese patients after undergoing bariatric surgery, but the underlying mechanisms remain unclear.

Aim:
to explore the associations between changes (Δ) in serum endocannabinoids [anandamide (AEA) and 2-arachidonoylglycerol (2-AG)] and Δ in ambulatory BP in morbidly obese patients 1 month after undergoing bariatric surgery (BxS).

Methods:
Thirty-one patients (25 female, 16 hypertensives, mean age 45±8,6 yr, mean body mass index [BMI] 44±5,6 kg/m
2
) with morbid obesity undergoing BxS were prospectively examined. Twenty-four hours-ambulatory BP parameters and serum AEA and 2-AG concentrations were evaluated at baseline and 1 month after BxS.

Results:
24h- systolic and diastolic BP decreased at 1 month (Table).

SBP=systolic blood pressure; DBP=diastolic blood pressure; BMI=body mass index; AEA=anandamide; 2-AG=2-arachidonoylglycerol

There were inverse correlations of Δ AEA with Δ 24h-SBP (P=-0,493; p=0,020) and with Δ 24h-DBP (P=-0,560; p=0.007). After adjusting for Δ BMI and baseline BP values, the correlation between Δ AEA and Δ 24h-DBP (β coef. =-0,38; 95%CI: -26,47 to -0,38) remained statistically significant (p=0,044; R
2
=0,523).

Conclusion:
in obese patients undergoing BxS, the decrease of 24h-DBP at 1 month is related to changes in AEA levels.
Abstract 3: Interaction Between NFκB and NFAT Coordinates Cardiac Hypertrophy and Pathological Remodeling
Authors: Liu Qinghang , Molkentin Jeffery
Published: March 2022

Both NFAT and NFkB are Rel homology domain-containing family members of transcription factors whose independent activities are critically involved in regulating cardiac hypertrophy and failure. Here we identified a novel transcriptional regulatory mechanism whereby NFkB and NFAT directly interact and synergistically promote transcriptional activation of one another in cardiomyoctes. We showed that NFkB-p65 co-immunoprecipitates with NFAT isoforms in cardiomyocytes, and this interaction is mapped to the Rel homology domain (RHD) within p65. Intriguingly, overexpression of p65-RHD disrupts the association between endogenous p65 and NFATc1, leading to reduced NFAT luciferase activity. Overexpression of p65-RHD or IKKβ leads to significant nuclear translocation of NFATc1, and expression of a constitutively nuclear NFATc1-SA similarly facilitated p65 nuclear translocation, further suggesting a physical interaction between p65 and NFATc1. Indeed, we observed that combined overexpression of p65 and NFAT leads to synergistic activation of NFAT transcriptional activity in cardiomyocytes. Conversely, NFAT transcriptional activity in cardiomyocytes is significantly reduced by inhibition of NFkB with IkBαM or dominant negative IKKβ. Importantly, hypertrophic agonist-induced NFAT activity is also significantly reduced in NFkB-p65 null MEFs compared to wild-type MEFs, while adenoviral-mediated expression of p65 restored NFAT luciferase activity in p65 null MEFs. In vivo, cardiac-specific deletion of NFkB-p65 using a Cre-LoxP system caused a ∼ 50% reduction in NFAT activity in NFAT-luciferase reporter mice. Moreover, ablation of p65 in the mouse heart decreased the hypertrophic response following pressure overload stimulation, reduced the degree of pathological remodeling, and preserved contractile function. Taken together, our results suggest a direct interaction between NFAT and NFkB pathways that may serve as a novel signaling mechanism in cardiac hypertrophy and failure.