bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P192: Psychosocial Correlates of Adherence to Non-pharmacological Therapy: An Examination of Baseline Results From the
SisterTalk Adhere
Intervention for Black Women
Authors: Kang Augustine W , Risica Patricia M
Published: March 2022

Significance:
Non-pharmacological treatment for blood pressure, such as dietary and Physical Activity (PA) adherence, is a key determinant of effective blood pressure management. Psychosocial variables such as self-efficacy, outcome expectations and motivation to engage in health behavior have been reported to be predictors of non-pharmacological adherence. However, few studies have studied these constructs among minority and at-risk populations such as hypertensive Black American women. Identifying psychosocial correlates of adherence is key to the development of efficacious interventions to improve adherence and overall health outcomes.

Methods:
We examined baseline measures (n = 323) of a sample of self-reported hypertensive Black women from
SisterTalk Adhere
, a randomized controlled trial to promote weight loss for hypertension control. Independent variables include (1) self-efficacy, (2) outcome expectations and (3) motivation for health behavior (further divided into PA and diet domain specific) Dependent variables are dietary and PA non-pharmacological adherence (operationalized as “none/some/most/all of the time” for diet and PA adherence in the past 4 weeks).

Results:
The mean age of the sample was 52.17±10.32. Regression analyses revealed that age was significantly associated with dietary adherence (
p
= .037). Dietary self-efficacy and positive outcome expectations from healthy eating were significantly associated with dietary adherence (both
p
< .001). Motivation for healthy eating was also significantly associated with dietary adherence (
p
= .043). Self-efficacy for PA and positive outcome expectations from PA were significantly associated with PA adherence (
p
= .006 and
p
< .001 respectively). Motivation for PA was not found to be associated with PA adherence.

Conclusion:
Results suggest that self-efficacy, outcome expectations and motivation for health behavior may be important targets to improve engagement in health behavior and non-pharmacological adherence among hypertensive Black women.
Abstract P193: Anti-fibrotic and Anti-oxidant Actions of a Muscadine Grape Extract Supplement on Hypertension-induced Cardiac Damage
Authors: Patil Pooja D , Westwood Brian M , Gallagher Patricia E , Tallant E. Ann
Published: March 2022

Approximately 75 million U.S. adults suffer from hypertension, an independent risk factor for pathological cardiac hypertrophy. The current study examines the effects of a muscadine grape extract formulation (MGE; Piedmont Research & Development Corp.) with anti-oxidant, anti-inflammatory, and anti-proliferative properties on hypertension-induced cardiac damage in Sprague-Dawley rats receiving a 4-week Ang II infusion (24 μg/kg/h). We previously showed that MGE in the drinking water had no effect on blood pressure or systolic function in normotensive or hypertensive rats; however, MGE ameliorated the Ang II-induced decrease in diastolic function and increase in cardiomyocyte cross-sectional area. In the current study, MGE supplementation of Ang II-treated rats decreased interstitial cardiac fibrosis, reducing collagen III staining in the myocardium (0.9 ± 0.2% control, 6.8 ± 1.0% Ang II, and 2.8 ± 0.4% Ang II/MGE; p<0.01). MGE alone had no effect on any parameters assessed. The positive correlation between collagen and diastolic function with Ang II compared to control (p<0.01) was significantly abrogated by co-administration of MGE. TGFβ is a critical cytokine that stimulates cardiac fibrosis. The Ang II-mediated increase in cardiac TGFβ mRNA was attenuated by MGE (relative gene expression: 1.0 ± 0.1 control, 2.0 ± 0.4 Ang II, 1.1 ± 0.2 Ang II/MGE; p<0.05) as was downstream pSmad2 protein in cardiomyocytes (% nuclei: 26.1 ± 6 control, 53.8 ± 9.6 Ang II, 27.6 ± 5.5 Ang II/MGE; p<0.05) and cardiac fibroblasts (2.3 ± 1.1 control, 11.7 ± 4.9 Ang II, 2.2 ± 0.5 Ang II/MGE; p<0.05). NADPH oxidase is a primary contributor to cardiac oxidative stress and excess ROS production can lead to activation of pro-fibrotic pathways. MGE significantly reduced the Ang II-induced increase in p22phox mRNA (relative gene expression: 1.0 ± 0.1 control, 1.9 ± 0.1 Ang II, 1.0 ± 0.1 Ang II/MGE; p<0.0001) and protein (relative density: 1.0 ± 0.2 control, 5.4 ± 1.6 Ang II, 2.9 ± 1.0 Ang II/MGE; p<0.05), a subunit required for NADPH oxidase activity. These results indicate that MGE ameliorates cardiac fibrosis by targeting NADPH oxidase and the TGFβ pro-fibrotic pathway, suggesting that MGE supplementation may be an effective, natural therapeutic for hypertension-induced cardiac damage.
Abstract P195: Digital Health Tool Use and Outcomes in People With Uncontrolled Hypertension
Authors: Senecal Conor , Widmer Robert J , Lerman Amir
Published: March 2022

Background:
Digital health applications are increasingly ubiquitous, however the effectiveness remains mixed. Using a large working age cohort we sought to evaluate use of a digital health intervention (DHI) in people with uncontrolled hypertension and their changes in blood pressure control.

Methods:
We analyzed usage of DHI and change in blood pressure control and other CVD risk factors over one year in 9,724 participants of a work health program across 81 organizations in 42 states. Patients were selected from a larger cohort based on a previous ICD-9 diagnosis of hypertension. Patients with uncontrolled hypertension, systolic blood pressure (SBP) greater than 140 mmHg, at the initial visit were included. Application users, defined as at least one login, were compared to non-users. Changes in blood pressure, weight, and lipids were compared at one year.

Results:
Users and non-users were similar in age, weight, blood pressure and lipids at baseline, though users were more likely to be female. At one year users had 16.44 mmHg reduction in SBP and 7.16 mmHg reduction in diastolic blood pressure (DBP), both statistically significantly more than non-users (Table 1). Both users and non-users had an improvement in LDL from baseline, while non-users also had an improvement in BMI and TG from baseline.

Discussion:
These results show that the use of a DHI is associated with improved systolic and diastolic blood pressure control compared to a workplace health program alone.
Abstract P203: pNaktide Targeted to Adipocytes Inhibits Na/k-atpase Reactive Oxygen Species, Systemic Inflammation, and Obesity Development in Mice Fed a Western Diet (WD)
Authors: Martin Rebecca , Brickman Cameron , Liu Jiang , Sodhi Komal , Shapiro Joseph I
Published: March 2022

Introduction:
Obesity is a worldwide epidemic with many comorbidities. It has been demonstrated that oxidative stress can exacerbate obesity development. We have previously published that systemic administration of pNaKtide, a Na/K-ATPase signaling antagonist, was able to decrease oxidative stress and adipogenesis by blocking Na/K-ATPase signaling mediated amplification of oxidative stress.

Hypothesis:
Adipocyte dysfunction in mice fed western diet (WD) may be prevented by lentiviral-mediated adipocyte-specific delivery of pNaKtide.

Methods:
C57Bl6 mice were randomly divided into five groups: 1) normal chow 2) normal chow+lenti-adipo-pNaKtide 3) WD 4) WD+lenti-adipo-GFP and 5) WD+lenti-adipo-pNaKtide (n=6-8/group). Lentiviral constructs with pNaKtide driven by an adiponectin promoter were used to achieve pNaKtide expression specifically in adipose tissue. Groups 2 and 5 were given an intraperitoneal injection of lenti-adipo-pNaKtide and group 4 was given an intraperitoneal injection of lenti-adipo-GFP at beginning of the experiment and again at week 2; total time=12 weeks. Body weight was measured weekly. Glucose clearance was determined using an intraperitoneal glucose tolerance test before termination of the experiment. At sacrifice body weight, visceral and subcutaneous fat content of all mice were measured. Blood samples were collected for determination of inflammatory cytokines. Tissues were flash frozen and maintained at -80°C.

Results:
Lenti-adipo-pNaKtide significantly reduced WD-induced weight gain, and visceral and subcutaneous fat content. Lenti-adipo-pNaKtide reduced WD-induced changes in glucose tolerance and inflammatory markers TNFα, IL-6 and MCP-1 (p<0.05). An increase in cardiac hypertrophy in WD animals was attenuated with lenti-adipo-pNaKtide (p<0.05). Visceral fat of WD mice expressed higher levels of adipogenic markers PPARγ, FAS, and C/EBP. WD-induced Na/K-ATPase signaling was decreased.

Conclusion:
Collectively this study introduces the novel idea that adipocytes may have a systemic effect. Specifically targeting pNaKtide to the adipocytes with lenti-adipo-pNaKtide ameliorates this systemic effect. This new information is important in the development of new therapeutic targets for obesity.
Abstract P209: Mechanistic Target of Rapamycin Complex 1 (mTORC1) Signaling Generates Reactive Oxygen Species in Mouse Endothelial Cells via NFkB Signaling
Authors: Reho John J , Guo Deng-Fu , Olson Andrew , Rahmouni Kamal
Published: March 2022

Obesity is associated with vascular endothelial dysfunction and reactive oxygen species (ROS) signaling. We have previously demonstrated a novel role for the mechanistic target of rapamycin complex 1 (mTORC1) signaling pathway in the regulation of cardiovascular reactivity. In this study, we assessed the ability of mTORC1 signaling to generate ROS in mouse endothelial cells (MEC) and investigated the role of NF
κ
B signaling as a potential transcriptional regulator. We used an adenoviral construct of a constitutively active (CA) S6-kinase (Ad-CAS6K) to enhance mTORC1 signaling in MEC. Infection of MEC (48hrs) with the Ad-CAS6K resulted in increased mRNA levels of NADPH oxidase 1 (3.4±0.7 fold) and 2 (3.6±0.6 fold) and decreased superoxide dismutase 2 expression (0.5±0.2 fold) compared to an adenoviral GFP (Ad-GFP) control (p<0.05). Next, we used dihydroethidium staining as a measure of ROS signaling in MEC. Infection of MEC with the Ad-CAS6K enhanced ROS signaling compared to the Ad-GFP (1.5±0.1 fold; p<0.05). Conversely, infection with a dominant negative S6K construct (Ad-DNS6K) decreased ROS generation (0.6±0.1; p<0.05). Notably, infection of aortic rings of wildtype mice with the Ad-CAS6K resulted in reduced acetylcholine induced vasorelaxation compared to Ad-GFP (Max. relaxation: 67±5 vs. 81±3%; p<0.05) with no change in relaxation evoked by sodium nitroprusside (Max. relaxation: 90±1% vs. 90±2%). The reduced endothelial-mediated vasorelaxation was rescued with Tempol (Max. relaxation: 74±2%; p<0.05) indicating ROS signaling as a mechanism of mTORC1 signaling induced endothelial dysfunction. Using immunoprecipitation assays, we uncovered a physical interaction between the mTOR subunit and the IKKβ subunit of the NF
κ
B complex. We also found that blockade of the IKKβ subunit with BMS-345541 (300nM) prevented the increased ROS generation in MEC in response to the Ad-CAS6K (1.4±0.1 vs 0.8±0.1 fold; p<0.05) implicating NF
κ
B in underlying mTORC1 signaling induced ROS generation. Our data demonstrate that enhanced mTORC1 signaling in endothelial cells impair endothelial function via ROS production and NF
κ
B signaling complex and may play a critical role in the endothelial dysfunction associated with various conditions including obesity.
Abstract P213: Metabolic and Cardiovascular Effects of Melanocortin-4 Receptors in Leptin Receptor-expressing Neurons
Authors: Rodriguez Cruz Eva N , Rahmouni Kamal , Cui Huxing
Published: March 2022

Studies in both humans and rodents have clearly established a role for melanocortin-4 receptor (MC4R) in energy homeostasis, sympathetic and cardiovascular function. Both MC4R-deficient humans and rodents develop severe obesity while keeping the sympathetic tone and blood pressure below or within the normal range, indicating that MC4R is required for obesity-associated sympathetic overdrive and hypertension. Indeed, sympathetic overdrive and cardiovascular side effect have been a major problem in developing anti-obesity drugs that target MC4R. Thus, better understanding the neural substrates that mediate distinct metabolic and sympathetic actions of MC4R is needed to provide a more specific target to treat obesity and associated hypertension. We have recently mapped the pattern of co-expression of MC4R and leptin receptor (LepR) and found that, throughout entire mouse brain, these two metabolically important receptors are uniquely co-expressed only in neurons of two brain regions, including the lateral hypothalamic area (LHA) and the periaqueductal gray (PAG). To specifically test the role of MC4R in LepR-expressing neurons, we generated mice with MC4R expression only in LepR-positive neurons by breeding Cre-dependently “reactivatable” MC4R-null mice (MC4R-TB) to LepR-Cre mice. We found that specific re-expression of MC4Rs only in LepR-positive neurons partially suppresses body weight gain especially in female mice (MC4R-TB 31.73 ± 2.2g vs. MC4R-TB::LepR-Cre 27.97 ± 1.8 g, p=0.0013). Direct multi-fiber recording of renal nerve activity (RSNA), however, revealed that neither baseline activity nor MC4R agonist-induced RSNA is significantly altered by restoration of MC4R signaling in LepR-positive neurons (change of RSNA in MC4R-TB 8.844 ± 3.05 vs. MC4R-TB::LepR-Cre 11.95 ± 2.7, p>0.05). Our results suggest that MC4R signaling in leptin-responsive neurons is sufficient to suppress body weight in female mice, but seemingly has no effect on sympathetic traffic to the kidney. Further investigation of the effect of MC4Rs in LepR-positive neurons on blood pressure regulation is underway.
Abstract P215: NADPH Oxidase Dependent Oxidative Stress and Redox Sensitive MAPKs Contribute to the Mechanisms of Acupuncture in Rostral Ventrolateral Medulla of Spontaneously Hypertensive Rats
Authors: Wang Xuerui , Yang Jingwen , Ji Caishuo , Zeng Xianghong , Shi Guangxia , Fisher Marc , Liu Cunzhi
Published: March 2022

NADPH Oxidase-Dependent Oxidative Stress and Redox-Sensitive MAPKs Contribute to The Mechanisms of Acupuncture in Rostral Ventrolateral Medulla of Spontaneously Hypertensive Rats.

Oxidative stress in the rostral ventrolateral medulla (RVLM), where the sympathetic nervous control center is located, contributes to neural mechanisms of hypertension. Acupuncture was previously reported to favorably affect high blood pressure. However, little is known about the effect of acupuncture on oxidative stress-modulated mechanisms in hypertension. This study was designed to evaluate the hypothesis that acupuncture exerts an antihypertensive effect via ameliorating oxidative stress and the redox-sensitive pathway in the RVLM of spontaneously hypertensive rats (SHRs). Two weeks of acupuncture reduced blood pressure and sympathetic nervous system activity in SHRs. Oxidative stress in the RVLM was alleviated by acupuncture, accompanied by a decrease in NADPH oxidase activity and expression of its subunits. Acupuncture significantly altered the mitogen-activated protein kinases (MAPKs) signaling pathway as assessed by pathway enrichment analysis in a gene chip assay. The phosphorylation of p38MAPK, extracellular signal-regulated protein kinase (ERK)1/2, but not Jun N-terminal kinase (JNK), were downregulated by acupuncture. Microinjection bilaterally of the superoxide dismutase mimetic tempol, NADPH oxidase inhibitor apocynin or diphenyleneiodonium chloride into the RVLM mimicked the antihypertensive effect of acupuncture. In contrast, the NADPH oxidase agonist TBCA abolished the beneficial effects of acupuncture. Furthermore, injection of capsaicin, or surgical sectioning of the sciatic nerve abolished the antihypertensive effect of acupuncture. We conclude that acupuncture decreases NADPH oxidase derived ROS and inactivates its downstream MAPKs in the RVLM to restore sympathetic vasomotor activity and decrease blood pressure. The neural transmission from the sensory sciatic nerve is involved in mechanism of acupuncture in treating hypertension.
Abstract P218: Role of UCP2 on Mitochondrial Dysfunction and Blood Pressure Regulation in the Renal Oxidative Stress-mediated Hypertension Associated With Dj-1 Depletion
Authors: De Miguel Carmen , Hamrick William C , Jagarlamudi Sudhar , Ma Xiaobo , Asico Laureano , Jose Pedro , Cuevas Santiago
Published: March 2022

DJ-1 and uncoupling protein 2 (UCP2) exert protective roles against mitochondrial (MT) oxidative stress.

DJ-1
-/-

mice have increased systolic blood pressure (BP) (+30±3% vs WT, n=6). This study determined the mechanisms involved in the oxidative stress-mediated hypertension due to
DJ-1
germline deletion. There were no differences in sodium excretion, renal renin expression, NADPH oxidase activity and serum creatinine between

DJ-1
-/-

and WT mice (n=5). However, renal expression of nitro-tyrosine was increased in

DJ-1
-/-

mice (+176.8±31% vs WT mice, n=5). Tempol, a radical scavenger, normalized the BP (tempol: 118±2% vs 100±1% vs WT, n=4) and renal malondialdehyde (tempol: 160±23% vs 109±15% vs WT, n=4) in

DJ-1
-/-

mice. Tempol-treated

DJ-1
-/-

mice had higher serum nitrite/nitrate levels than placebo-treated (172±30% vs WT, n=4). Heat shock protein mtHSP60 was increased in

DJ-1
-/-

mice (2.9±0.1-fold increase vs WT, n=4), indicating MT stress. However, there were no changes in the renal mRNA expression of mitophagy, MT fusion and MT biosynthesis markers indicating that MT function was not altered. Renal expression of
UCP2
was increased in

DJ-1
-/-

mice (4.1±1.1-fold change vs WT, n=4), and was partially normalized by tempol (1.8±0.2-fold change vs WT, n=4),
UCP2
may have a protective role on MT function in this model.
UCP2
was selectively silenced via sub-capsular infusion of
UCP2
siRNA in the kidney (WT: 63%±7 vs control:

DJ-1
-/-

:60%±6 vs control; n=4). mRNA expression of mitophagy markers BNIP3 (-0.65±6-fold) and PINK1 (1.55±0.3-fold), MT fusion markers FIS1 (-0.29±0.03-fold) and NFN2 (1.42±0.06-fold), and MT biosynthesis marker PPRC1 (1.71±0.07-fold) were altered by
UCP2
silencing in

DJ-1
-/-

mice (n=4). Renal-selective silencing of
UCP2
normalized BP in

DJ-1
-/-

mice (

DJ-1
-/-

mice: 122±5 vs 98±7 mmHg, n=4), and the serum nitrite and nitrate concentrations (-40±9% vs WT, n=4). In conclusion, deletion of
DJ-1
leads to oxidative stress-induced hypertension associated with down-regulation of NO synthesis.
UCP2
has protective properties against the development of MT dysfunction in MT oxidative stress conditions
.
However, excessive and chronic over expression of
UCP2
could have deleterious consequences on BP regulation.
Abstract P220: Perception of Blood Pressure Control and Satisfaction with Hypertension Care in the Rural Setting
Authors: Bejjanki Harini , Gong Yan , Willis David , Siler-Marsiglio Kendra , Handberg Eileen , Haight Daniel , Smith Steven , Pepine Carl , Cooper-DeHoff Rhonda
Published: March 2022

Background:
National Health and Nutrition Examination Survey (NHANES) data indicate only 50% with hypertension (HTN) have controlled blood pressure (BP) yet limited data are available regarding patient’s knowledge of and satisfaction with their BP control.

Methods:
We utilized MyHealthStory, an online health information exchange portal to identify a rural, diverse HTN population. Adult patients (≥ 18 yrs) were recruited from existing federally-qualified health center (FQHC) sites. The survey was sent via email to 5000 patients who voluntarily shared email addresses through MyHealthStory. Equal numbers of men and women of any race/ethnicity with a diagnosis of HTN and who consented to receive information about research in MyHealthStory were targeted. We developed the 21 question survey using Qualtrics. The first 300 patients who completed the survey were included. We compared a) patient-reported BP control (BP ≥ 140/90; or <140/90) and patient-reported satisfaction with BP control (yes=happy, no=unhappy) and b) patient reported BP control and perception of BP control (well controlled, uncontrolled); using McNemar’s test for paired nominal data.

Results:
Here we report data from the 238 completed surveys received from Caucasians (n=184, 61% female) and African Americans (n=54, 83% female). Among 117 Caucasians reporting BP, 51% (n=60) reported uncontrolled BP, 52% (n=61) reported perception of well controlled BP (p=0.49). A total of 77% (n=46) of patients who had uncontrolled BP reported satisfaction with their HTN care compared to 96% in those who had controlled BP (
P
<0.0001). Among 32 African Americans reporting BP, only 13% (n=4) reported controlled BP but 31% (n=10) reported perception of well controlled BP (p=0.034). Among the 4 African Americans whose BP was controlled, all 4 reported satisfaction with their HTN care, while among the 28 patients who reported uncontrolled BP, 23 (82%) reported satisfaction with their HTN care (
P
<0.0001).

Conclusions:
Among hypertensive individuals, our survey data suggest there is a substantial disconnection between reported BP control, perception of BP control and satisfaction with HTN treatment, suggesting need for improvement in patient education surrounding HTN treatment and BP control.
Abstract P222: Prevalence of Hypotension in Adolescents
Authors: Bhatia-Barnes Jyoti , Gupta Monesha , Bell Cynthia , Samuels Josuha
Published: March 2022

Background:
Hypotension is a widely used but poorly defined clinical entity. Currently accepted definitions of hypotension are the Pediatric Advanced Life Support (PALS) definition, the Brain Trauma Foundation definition, and the International Pediatric Sepsis Consensus Conference. However, its prevalence has not been defined.

Objective:
To determine the prevalence of hypotension in school aged children.

Study Design:
This study is a retrospective analysis of data from a series of cross-sectional school-based blood pressure screenings completed by the Houston Pediatric and Adolescent Hypertension Program (HPAHP) at the UTHealth at Houston. Screenings were performed in 21 urban and suburban middle and high schools in the Houston area from 2001 to 2012. We restricted the data set to include subjects with three systolic blood pressure (SBP) measurements performed using a Spacelabs 90217 oscillometric monitor, who did not report taking hypertensive medication, and were not missing demographic information. We defined hypotension based on 2 criteria: average 1
st
-3
rd
SBP less than the 5
th
percentile for height, age and gender. Within these definitions we also restricted the hypotension definition to subjects with average SBP below 90mm Hg. Stata 13 SE was used for all data analysis.

Results:
Of the total 22, 382 children enrolled, 15,114 children met inclusion criteria: 52% females, mean age 13.6 years (range 10.1-19.9). Racial distribution reflected that of Houston: 35% White, 32% Hispanic, 23% Black, 8% Asian, and 2% other/unknown. Seventy one (0.5%) children had an average SBP reading less than the 5
th
percentile and 112 (0.7%) had an average SBP less than the 5
th
percentile or 90 mm Hg. Compared to students without hypotension, students with hypotension defined by only the 5
th
percentile were older (14.3 vs 13.6 years, t-test p=0.0001); they also tended to be more female, though not significantly (61% vs 39%, Chi
2
p-value=0.134).

Conclusion:
Although not as prevalent as hypertension in this age group (generally 2-5%), hypotension is a clinically significant entity that is seen in 0.5-0.7% of the general school-aged population.
Abstract P223: The ω-oxidation Pathway Underlies Hexadecanedioate Induced Blood Pressure Elevation
Authors: Alharbi Nora H , McBride Martin W , Padmanabhan Sandosh , Graham Delyth
Published: March 2022

Background:
A novel pathway for blood pressure (BP) regulation involving the dicarboxylic acid hexadecanedioate (HEXA) was identified in a study associating BP and mortality outcomes with fasting blood metabolites. The functional role of HEXA on BP elevation and vascular reactivity was confirmed by oral administration of hexadecanedioic acid to WKY rats. This study characterized the metabolic effects of hexadecanedioic acid administration in WKY rats, and assessed hemodynamic changes after modulation of endogenous HEXA levels by perturbing the ω-oxidation pathway in the SHRSP rat.

Methods:
WKY were treated orally with hexadecanedioic acid (250 mg/kg/day) or vehicle (n=5/group) for 3 weeks. Aorta, heart, brain, adipose, kidney, and liver were harvested and global metabolic profiles analysed by UPLC-MS/MS (Metabolon). The aldehyde dehydrogenase (ALDH) inhibitor disulfiram (25 mg/kg/day) or vehicle were administered to SHRSP rats for 14 days (n=7/group). BP was assessed by tail plethysmography and mesenteric arteries were used to assess vascular function by wire myography.

Results:
Treatment with hexadecanedioic acid increased HEXA levels in all tissues except brain in WKY rats. Metabolomic analysis identified increased fatty acid (FAs) metabolites (e.g. acylcarnitines) in heart, long-chain FAs and ketone body (β-hydroxybutyric acid) in kidney, and dicarboxylate FAs in adipose tissue from hexadecandioic acid treated rats indicating an impairment of FA β-oxidation and a shift towards ω-oxidation. SHRSP rats showed a significant reduction in BP after treatment with disulfiram (ΔSBP, 20.7±4.6 mmHg;
P
=2.4x10
-4
). Mesenteric resistance arteries from disulfiram treated SHRSP demonstrated a shift to the right in the contractile response curve to noradrenaline, indicating reduced vascular sensitivity compared to control vessels.

Conclusion:
Administration of hexadecanedioic acid in addition to increasing BP impacts several metabolic readouts including changes in FA metabolism. Importantly, we demonstrate that inhibition of the ω-oxidation pathway enzyme, ALDH, results in a significant lowering of BP in the hypertensive SHRSP rat. The ω-oxidation pathway is a target for further research to elucidate mechanisms by which HEXA affects BP.
Abstract P227: Prostaglandin EP
4
Receptor Mediate Angiotensin II-induced Hypertension via Activation of PRR/ENaC Pathway
Authors: Lu Xiaohan , Wang Fei , Yang Kevin , Peng Kexin , Yang Tianxin
Published: March 2022

Activation of COX-2/EP
4
pathway is shown to upregulate renal (pro) renin receptor (PRR) contributing angiotensin II (Ang II)-induced hypertension but the underlying mechanism remains unclear. Here, we tested the hypothesis that activation of EP
4
receptors increases ENaC expression/activity through PRR in the setting of Ang II treatment. Sprague-Dawley rats were infused for 2 weeks with vehicle, Ang II alone (100 ng/kg/min), or in combination with an EP
4
antagonist ONO-AE3-208 (ONO) (0.2 mg/kg/d). Following 14-d AngII treatment, 24-h urine volume and 24-h urinary Na
+
excretion both exhibited a significant increase which was completely blocked by ONO. Ang II infusion selectively elevated α-ENaC protein abundance (1.9-fold) in the renal medulla but not in the renal cortex, which was completely blocked by ONO, as assessed by immunoblotting. Interestingly, Ang II infusion elevated the cleaved γ-ENaC protein in both medulla and cortex, which was similarly abolished by ONO. In response to Ang II infusion, the increases in renal medullary mRNA were 1.7-fold for α-ENaC, 1.3-fold for β-ENaC, 2.4-fold for γ-ENaC as assessed by qRT-PCR; in contrast, none of the ENaC subunits showed obvious changes in the renal cortex. Ang II-induced increases in α-ENaC mRNA were reduced 41% by ONO with a modest effect on the response of β-ENaC and γ-ENaC. In cultured mpkCCD cells, Ang II treatment at 500 nM for 24 h induced a 2.5-fold increase of ENaC activity as assessed by epithelial volt-ohmmeter, which was completely blocked by ONO (10 μM) or a specific PRR inhibitor PRO20 (1.0 μM). To further determine the role of the EP4 receptor in the regulation of ENaC activity, the mpkCCD cells were treated with vehicle, CAY10580 alone (agonist of the EP
4
receptor, 10 μM), or in combination with PRO20 for 24 h. The ENaC activity showed a 2.0-fold increase in response to CAY10580, which was nearly completely abolished by PRO20 (n = 6-8, p<0.05). CAY10580 induced a 2.1-fold increase in PRR protein abundance. Overall, these results suggest that activation of EP
4
induces PRR, leading enhancement of ENaC expression/activity, eventually contributing to increased blood pressure in response to Ang II treatment.
Abstract P228: Chronically Enhanced Fructose Intake Induces Salt Sensitive Hypertension in Mice and Increases NKCC2 Phosphorylation
Authors: Kassem Kamal M , Harding Pamela , Ortiz Pablo
Published: March 2022

High fructose consumption is implicated in hypertension and renal damage in humans. The mechanisms for the renal effects of fructose are not clear. In normal rats, a high fructose diet (20%) does not increase blood pressure, but rapidly (one week) induces a salt sensitive increase in blood pressure. NaCl reabsorption by the thick ascending limb (TAL) is mediated by the NKCC2 cotransporter. Higher NKCC2 activity is involved in salt sensitive hypertension, and its phosphorylation is increased by fructose in rats. We hypothesized that high fructose (20% drinking water), would induce NKCC2 phosphorylation and induce salt sensitive hypertension in normal C57/Bl6J mice. To test this, mice were given 20% fructose in drinking water and after two weeks they were treated for 10 more weeks with either fructose alone or fructose plus 4% Na diet. Tail cuff systolic blood pressure (SBP) was measured after 3 weeks of training. Total NKCC2 expression, GAPDH and NKCC2 phosphorylation at Thre96/101 (P-NKCC2) were measured by Western blot in freshly isolated TALs. Baseline SBP was 112 ± 3 mmHg. A transient increase to 125± 4 mmHg (
p
<0.05) was observed after 1 week on fructose which later decreased to 119 ± 3 mmHg (
p
=0.14) by week 2. At the end of week 2, high salt chow was added. BP remained normal for 3 weeks and then increased to 137 ± 11 mmHg (
p
<0.05) by week 4 on high salt, but not in fructose alone mice. After 10 weeks SBP remained elevated in the fructose plus high salt diet group (124 ± 3 vs 110 ± 5 mmHg;
p
<0.05). Water intake and urine volume was similar between the two high fructose groups. After 10 weeks, fasting glucose was higher in all mice with fructose (fructose + HS: 231± 8, fructose: 218 ± 7, normal diet: 122 ± 4 mg/dL,
p
<0.001). A fructose diet increased P-NKCC2 by 248 ± 82% (n=5,
p
<0.01) by one week and by 61 ± 21% (
p
<0.05) at week 12. The combination of high-fructose high salt diet increased P-NKCC2 by 226 ± 75% (
p
<0.05) at 12 weeks. Transcriptome profiling of isolated mouse TALs showed that 20% fructose (7 days) increased the expression of genes upstream of the SPAK and AMPK pathway. We conclude that 20% fructose in drinking water leads to a salt dependent increase in SBP, which appears to occur after mice develop abnormalities in glucose metabolism.
Abstract P230: Uncoupling of NOS3 is Involved in Diminished Flow-induced No Production in Dahl Salt-sensitive Rat Thick Ascending Limbs
Authors: Hong Nancy J , Gonzalez-Vicente Agustin , Garvin Jeffrey L
Published: March 2022

Increased luminal flow enhances nitric oxide (NO) production in thick ascending limbs (TALs). NO produced by NO synthase 3 (NOS3) inhibits Na transport. However, its effect on transport is reduced in Dahl salt-sensitive (SS) vs salt-resistant rats (SR). In Sprague-Dawley rat TALs, angiotensin II can acutely cause NOS3 to uncouple to produce superoxide (O
2
-
) thereby reducing NO production. We hypothesized that flow-induced NO production is decreased in SS TALs and that this is due to NOS3 uncoupling. We measured flow-induced NO in isolated perfused TALs using the fluorescent dye DAF-FM and performed Western blots of renal medullary lysates. Flow-induced NO production was reduced 69% in TALs from SS (11±2 arbitrary units (AU)/min, n=6) vs SR (35±6 AU/min, n=8, p < 0.008). This difference between strains was not due to altered NOS3 expression (NOS3/GAPDH ratio of 0.91 ± 0.08 for SS vs 1.09 ± 0.08 for SR; n = 5 for each). The difference in flow-induced NO between strains was slightly reduced in the presence of the superoxide (O
2
-
) scavenger tempol (19±2 vs 30±5 AU/min for SS and SR, respectively; n=9 for each strain, p < 0.04), suggesting that scavenging of NO by O
2
-
plays only a minor role in the difference in flow-induced NO production between SS and SR thick ascending limbs. We next investigated whether NOS3 uncoupling could account for the difference between strains by using the fluorescent dye dihydroethidium to measure flow-induced O
2
-
before and after treatment with the NOS inhibitor L-NAME. Blocking NOS3 reduced O
2
-
production in SS TALs by 21±7%, from 38±5 to 30±5 AU/min (n=6, p < 0.05) whereas it had no effect in SR TALs (26±6 vs 28±3, n=5). We conclude that the diminished flow-induced NO in SS TALs is not due to differences in NOS3 expression nor acute flow-induced O
2
-
, but rather in large part due to uncoupling of NOS3. Impaired flow-induced NO production in TALs could contribute to the Na retention associated with salt-sensitive hypertension.
Abstract P235: Renin and Prorenin Uptake by Human Conditionally Immortalized Proximal Tubule Epithelial Cells Involves Megalin and Mannose 6-phosphate Receptor
Authors: Martini Alexandre G
Published: March 2022

Renin is filtered by the glomerulus, and subsequently reabsorbed in the proximal tubule in a megalin-dependent manner, as evidenced by a 20-40-fold rise in urinary renin in patients with Dent’s disease or Lowe syndrome, i.e. disorders characterized by defective proximal tubular reabsorption. Remarkably, the reabsorption of filtered prorenin appears to be complete, as it can only be detected in urine of patients with Dent’s disease or Lowe syndrome. To further investigate megalin-mediated renin and prorenin reabsorption, human conditionally immortalized proximal tubule epithelial cells (ciPTEC) were incubated at 37 °C or 4 °C with recombinant human renin or prorenin (1000 U/L), in the absence or presence of 10 mmol/L mannose 6-phosphate (M6P) to block (pro)renin binding and internalization via M6P receptors. Cell lysate (pro)renin levels were measured by immunoradiometric assay both before and after prorenin activation. At 37 °C, cellular renin and prorenin tended to level off at 6 hours, reaching lysate levels of 75±18 and 100±54 pg/mg (mean±SEM), and all prorenin was detected as renin. Incubating ciPTEC at 4 °C diminished the cellular accumulation of renin and prorenin by 90%, and at this temperature prorenin remained in its inactive conformation. M6P blocked renin and prorenin uptake by >80% under all conditions. Importantly, M6P also concentration-dependently blocked the uptake of fluorescently labeled albumin (IC
50
= 3 mmol/L), to the same degree as the megalin inhibitor RAP-GST (IC
50
= 50 μg/mL). Since albumin is internalized in a megalin-dependent manner, these data indicate that M6P, on top of blocking M6P receptors, also interferes with the megalin-dependent uptake process. In conclusion, ciPTEC bind and internalize renin and prorenin in a temperature-dependent manner. This process involves megalin and/or M6P receptors, and results in prorenin activation at 37 °C.
Abstract P237: The At2 Receptor Agonist, C21, Can Also Stimulate Mas and Mrgd Receptors
Authors: Tetzner Anja , Naughton Maura , Gebolys Kinga , Sala Esther , Villacañas Óscar , Walther Thomas
Published: March 2022

Background:
It is well accepted that Compound 21 (C21) is a highly selective non-peptide angiotensin AT2 receptor agonist. C21 as well as angiotensin (Ang)-(1-7) have cardiovascular protective effects and are opponents of the often detrimental Ang II within the renin-angiotensin system. Since the chemical structure of C21 is similar to the Mas receptor specific non-peptidic agonist AVE0991, and we could recently show that the AT2 receptor blocker, PD123319, can also block the effects of Ang-(1-7) at its natural receptors, Mas and MrgD, we tested whether C21 is also not AT2-specific but can stimulate the two Ang-(1-7) receptors, too.

Methods and Results:
Using cAMP as readout in receptor-transfected HEK293 cells, we generated pharmacological proof that Mas (EC
50
= 1.995 x 10
-12
M) and MrgD (EC
50
= 2.958 x 10
-9
M) are functional receptors for C21, whereby the three receptor blockers, A779, D-Pro7-Ang-(1-7), and PD123319 showed receptor-specific effects towards C21 signalling. Furthermore, C21 elevated the cAMP concentration in primary cells such as mesangial cells (EC
50
= 1.12 x 10
-6
M). However, significant increase in cAMP levels, but not in PKA activity, was still detectable in mesangial cells isolated from AT2-deficient mice, but completely blunted in Mas/MrgD-double knockouts. Finally,
in silico
modelling was performed to illustrate the structural similarities and differences between C21, AVE0991, and Ang-(1-7).

Conclusions:
Our results identify C21 as not being a specific AT2 receptor agonist but also interacting with the two Ang-(1-7) receptors, Mas and MrgD. Therefore, the partial overlap in beneficial effects of Ang-(1-7) and C21 might be based on the stimulation of the same receptors under specific pathophysiological circumstances. This also enforces the revisit of such publications which concluded on AT2 function by only using C21.
Abstract P243: Smooth Muscle Cullin-3 Deficiency Causes Severe Early Onset Hypertension and Nitric Oxide Resistance
Authors: Agbor Larry N , Nair Anand , Wu Jing , Davis Deborah R , Keen Henry L , Quelle Frederick W , McCormick James A , Singer Jeffrey D , Sigmund Curt D
Published: March 2022

Mutations in
Cullin-3
(
Cul3
) resulting in exon 9 skipping (CUL3α9) cause human hypertension. We demonstrated that selective expression of CUL3α9 in smooth muscle causes arterial stiffness and hypertension. We hypothesized that deletion of CUL3 in smooth muscle causes severe hypertension. Mice carrying a conditional allele of CUL3 were bred with mice expressing a tamoxifen-inducible CRE-recombinase driven by a smooth muscle promoter. Mice were administered tamoxifen
i.p.
(75 mg/kg) for 5 consecutive days to generate smooth muscle CUL3 knockout (S-CUL3KO). CUL3 protein was undetectable whereas Cullin-1 protein was preserved in aorta from S-CUL3KO mice. We assessed vascular function in the cerebral basilar artery and aorta using pressurized and wire myograph, respectively. Blood pressure (BP) was measured by radiotelemetry. S-CUL3KO mice exhibited significantly increased systolic BP (SBP) at 2 weeks and 4 weeks post tamoxifen compared to corn oil controls (2 wks SBP mmHg: 145±1 vs 115±2, p<0.001; 4 weeks SBP: 169±1 vs 115±3, p<0.001). Pulse wave velocity was also increased in S-CUL3KO mice (3.7±0.1 m/s vs 2.2±0.1, P<0.001), suggesting increased arterial stiffness. Aorta from S-CUL3KO mice exhibited severely impaired vasorelaxation to acetylcholine (ACh) compared to controls (at 100 μM: 1.0±3% vs 77±5%, p<0.0001), and to the nitric oxide donor sodium nitroprusside (SNP) (at 100 μM: 15±4% vs 96±1%, p<0.001). In agreement with data from aorta, cerebral basilar artery from S-CUL3KO mice also exhibited significant impairment to ACh- and SNP-mediated vasorelaxation. Conversely, S-CUL3KO aorta retained an ability to relax in response to a cGMP analogue (8-pCPT-cGMP, at 100 μμ: 8-pCPT-cGMP 70±3%) and to the heme-independent soluble guanylate cyclase activator (BAY 58-2667, at 10 μμ: BAY 58-2667 94±2%), indicating that downstream mechanisms controlled by cGMP remain intact in the absence of CUL3. Captopril (120 mg/kg/day) was sufficient to normalized BP in S-CUL3KO mice to pre-tamoxifen levels (change in SBP mmHg: -50.2±1.9 S-CUL3KO vs -25.0±2 control, P<0.0001). We conclude that smooth muscle CUL3 is a major BP determinant, and identifying novel CUL3 substrates in smooth muscle would be beneficial as therapeutic targets in the treatment of hypertension.
Abstract P250: Comparison of Clinical Variables Independently Associated With Large Artery Stiffness and Microvascular Dysfunction in African-American Diabetic Patients
Authors: Eluwole Ayobami , Adedayo Ajibola , Tedla Fasika , Kremer Arye , Mastrogiovanni Nicole , Afzal Mohammed , Rosenberg Carl , Dreizen Paul , Larossa John , Boutjdir Mohamed , Banerji Mary Ann , Brown Clinton , Salifu Moro , Bakilah Ahmed , Lazar Jason
Published: March 2022

Rationale:
Diabetes mellitus is a risk factor for large and small vessel disease and portends a disproportionate morbidity and mortality burden in African Americans. Diabetes is often accompanied by cardiovascular risk factors that predispose to arteriopathy, it’s been shown to impair large and small vessel function prior to onset of target organ damage. Factors contributing to subclinical vascular dysfunction have not been well studied in this population. The objective of this study was to compare clinical variables associated with large artery stiffness and microvascular dysfunction.

Methods:
A total of 141 patients with diabetes were recruited from medical clinics over a 6 month period. Medical information was obtained via interview and medical record review including laboratory results. Pooled Cohort Score was calculated for each subject. Microvascular function was assessed by vascular reactivity index (VRI), which assesses changes in digital temperature before and after release of arterial cuff occlusion (VENDYS 5000BC DTM system (Endothelix, Inc.). Large artery stiffness was assessed by carotid-femoral pulse wave velocity (PWV) using applanation tonometry (Sphygmocor, Atcor Inc.).

Results:
Mean age was 60+8 years, 64% were female. 80% had hypertension, 90% had dyslipidemia and 15% had chronic kidney disease. Mean HbA1C levels were 8.1+2.2%. On univariate analysis, Pooled-cohort-score was significantly correlated with PWV (r=.25, p=.003) but not with VRI (r=.02, p=.78). Neither PWV nor VRI was significantly correlated with HbA1c. On multivariate analysis, PWV was independently associated with age, gender, creatinine level and waist circumference but not with traditional risk factors (R2=.26, p<.001 for model). VRI was not significantly correlated with any of the clinical or laboratory measures.

Conclusions:
Large artery stiffness is independently associated with age, female gender, renal function and waist circumference but not with poorer glycemic control. In contrast, microvascular dysfunction was not associated with any of the clinical or laboratory measures. The role of metabolic risk factors in the differential progression of subclinical large and small artery vessel dysfunction and outcomes merit further study.
Abstract P252: Hemochromatosis Induce Arterial Stiffness and Endothelial Dysfunction and They Can Be Effectively Reversed by Phlebotomy
Authors: Duarte Mariano , Aquieri Analia , Coyle Javier , Gonzalez Ballerga Esteban , Savio Galimberti Eleonora , Reyes Toso Carlos
Published: March 2022

Hereditary Hemochromatosis (HH) is a genetically determined disease where iron is excessively stored in tissues, causing organ injury and failure. An increase of cardiovascular risk (CVR) has been reported in HH. Part of the iron overload induces vascular fibrosis and endothelial dysfunction (ED). Therefore, arterial wall emerge as a novel organ target organ in HH. Carotid-femoral pulse wave velocity (PWV-CF) assesses arterial stiffness (AS), and has predictive value for CV events. PWV variation pre- and post-induced transient ischemia of brachial artery (PWV-CB) has been validated to evaluate ED. We hypothetized that patients with HH have an increase in PWV-CF and PWV-CB due to iron overload, and that phlebotomy by reducing iron levels decreases arterial stiffness and restores endothelial function. To test this hypothesis we assessed PWV-CF and PWV-CB in 52 patients with HH before and after phlebotomy and compare against control. PWV-CF and PWV-CB variations were assessed using Complior System (Artech-Medical, Francia). PWV-CF was significantly increased in HH patients compared to control group (8.5±1.7 m/s vs. 6.4±0.8 m/s, P<0.001, n=52/30 patients/group), suggesting increased AS. Changes in PWV-CB after transient ischemia were reduced in HH patients (0.3%) compared to controls (8.2%) (P<0.001), suggesting ED. Phlebotomy significantly reduced PWV-CF in HH patients (Post-phlebotomy: 6.5±0.8 m/s vs. Pre-phlebotomy: 8.5±1.7 m/s, P<0.001) back to control values (6.4±0.8 m/s). Changes in PWV-CB variation were partially reduced post - phlebotomy. (PWV-CB variation was 0.3 % in HH pre-phlebotomy vs. - 5.7 % in HH post- phlebotomy). These results showed a significantly increased AS and ED in HH group suggesting that arterial wall represents a novel target in HH and could may the nexus to cardiovascular disease. Both parameters (AS and ED) were significantly improved after phlebotomy, Our findings supports future research including more patients with HH and others iron overload disorders.
Abstract P257: Investigating Angiotensin II Infusion as a Model of Superimposed Preeclampsia in Pregnant Stroke Prone Spontaneously Hypertensive Rats
Authors: Morgan Hannah L , Butler Elaine , Beattie Elisabeth , McBride Martin W , Graham Delyth
Published: March 2022

Background:
Hypertensive disorders of pregnancy are a leading cause of maternal mortality worldwide. Chronic hypertension with superimposed preeclampsia is becoming more prevalent in modern society. Animal models are vital for understanding this condition. Our aim was to induce a preeclamptic phenotype in pregnant stroke prone spontaneously hypertensive (SHRSP) rats by angiotensin II (AngII) infusion.

Methods:
AngII, at a dose of 500ng/kg/min or 1000ng/kg/min, was administered by minipump in pregnant SHRSP rats from gestational day (GD)10. Cardiovascular and renal changes were monitored using radiotelemetry, echocardiography and weekly urine collections. GD18
ex vivo
uterine artery function and structure were assessed using myography.

Results:
Both AngII doses resulted in an immediate and sustained increase in systolic blood pressure compared to vehicle control (
172.5±5.0mmHg; 500ng/kg/min and 204.1±5.6mmHg; 1000ng/kg/min vs 159.0±0.4mmHg; p<0.001
). Cardiac output increased over pregnancy in the vehicle group (
53±6L/min vs 81±5L/min; p<0.01
), however this trend was not observed in 500ng/kg/min treatment group (
60±6L/min vs 56±11L/min
) and was reversed in 1000ng/kg/min treatment group (
60±6L/min vs 36±6L/min; p<0.01
). GD14 urinary albumin:creatinine ratios were significantly increased in both treatment groups (
1.3±0.2; 500ng/kg/min, 2.9±0.1; 1000ng/kg/min vs 0.5±0.2; vehicle; p<0.05, p<0.001
). Uterine arteries in the 1000ng/kg/min treatment group demonstrated significantly reduced cross-sectional area (

4.8x10
4
±32μm
2
vs 7.6x10
4
±16μm
2
; p<0.01

). Both treatment groups had a reduced vasorelaxation to 2x10
5
M carbachol (
93.6±3.9%; 500ng/kg/min, 84.7±4.3%; 1000ng/kg/min vs 56.9±21.1%; vehicle; p<0.05
) and contractile response to 2x10
5
M noradrenaline demonstrated an increased trend in both treatment groups (
66.9±16.1KPa; 500ng/kg/min, 61.8±6.2KPa; 1000ng/kg/min vs 45.1±5.7KPa; vehicle
).

Conclusion:
This study demonstrates that AngII infusion in pregnant SHRSP rats can mimic specific haemodynamic, cardiac and urinary profiles common to preeclamptic women. This rodent model of superimposed preeclampsia will be crucial for detailed investigation of underlying causes and treatment options for this condition.