bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P454: Does Intensive Pulse Pressure control results in lower rates of cardiovascular events? An Analysis of Systolic BP Intervention Trial (SPRINT) database
Authors: Tran Quan
Published: July 2021

Introduction:
A wide pulse pressure (PP) is a marker of increased artery stiffness and high cardiovascular risk. A high PP may reflect already diseased arterial walls, with several adverse cardiac implications of potential prognostic value. Previous studies show that arterial stiffness is itself an independent predictor of the risk of all-cause mortality, cardiovascular mortality, coronary events and stroke in hypertensive subjects. No guidelines exist regarding tight pulse pressure in chronic hypertension population.

Method:
For the purpose of our study, we examined SPRINT data collected between November 2010 and August 2015. The role of PP and DBP was studied as a qualitative parameter with separation according to the 4 quartiles defined in the studied population. We then combined the results of both PP and DBP in conjunction with preexistence intensive vs. control arm for further analysis.

Results:
Our secondary analysis of SPRINT data first confirmed NEJM article that intensive SBP control reduced rates of heart failure (HF), CVD death, Death from any cause, and SPRINT primary outcome or death. There was a significant difference in primary outcome in each PP quartile. Higher PP quartiles were more likely to have higher rate of MI, stroke, HF, CVD death, and SPRINT primary outcome or death. There also a significant increase in SPRINT primary outcome and Heart Failure when DBP <70 in the intensive treatment arm. Finally, SPRINT data has confirmed that participants in PP
1
and PP
2
with DBP > 70 have significant lower rate of SPRINT primary outcome and death compared to other group regardless of treatment arm.

Conclusion:
In the SPRINT database of 9347 participants, we found that higher PP conferred an increased risk for multiple adverse cardiovascular event. Our study is among the largest US studies of PP and adds further support to the prognostic utility of PP. In addition, the SPRINT database offers a contemporary analysis of the relationship between PP and DBP, which is important given that nature pathophysiology of blood pressure. Current study also strengthen the possibility that DBP <70 mm Hg was associated with a higher rate of cardiac events.
Abstract P461: The Association of NOV/CCN3 With Obstructive Sleep Apnea in Severe Morbid Obesity: Preliminary Evidence of a Novel Biomarker in OSA
Authors: Weingarten Jeremy A , Bellner Lars , Peterson Stephen J , Zaw Moe , Chadha Puja , Mehta Hemal H , Abraham Nader G
Published: July 2021

Background:
Obstructive sleep apnea (OSA) is strongly associated with cardiovascular and metabolic abnormalities, although the mechanism driving this association is not well understood. NOV/CCN3, a multifunctional extracellular matrix protein, may play a mechanistic and/or prognostic role in these associations. We hypothesized that patients with OSA will have increased levels of NOV, and that NOV can serve as a biomarker in patients to predict OSA as well as metabolic and cardiac risk.

Materials and Methods:
10 morbidly obese subjects presenting to the sleep laboratory for clinical evaluation of possible OSA and 10 healthy lean controls underwent overnight polysomnography and clinical evaluation. Blood samples were analyzed for NOV levels, adiponectin, and IL-6.

Results:
OSA was found in 9 obese subjects and 3 lean subjects. NOV levels were significantly higher in the obese vs lean group (2.2±0.3 vs 1.4±0.2 fold change, p<0.03) while apnea-hypopnea index (AHI) was similarly higher in the obese vs lean group (22.9±6.7 vs 8.4±4.1 events/hr, p<0.03). NOV and AHI were positively correlated (ρ=0.49, p=0.033). IL-6 was elevated while adiponectin was reduced in obese vs lean.

Conclusions:
NOV levels and AHI, a measure of OSA severity, were higher in obese subjects vs lean controls and were correlated with each other. NOV may be a marker of OSA severity and a biomarker for cardiovascular and metabolic disease in OSA patients.
Abstract P462: MiRNA and Biomarkers of Metabolic Syndrome: Correlating Biomarkersfor Early Detection of Metabolic Syndrome in Obese Females in West Virginia
Authors: Lakhani Hari Vishal , Sharma Dana , Nawab Athar , Sodhi Komal
Published: July 2021

Metabolic syndrome, a multifactorial disease, causes complications like cardiovascular disease and diabetes mellitus. As metabolic syndrome develops, altered levels of cytokines and miRNAs are measured in the circulation. We constructed a panel detecting abnormal levels of cytokines and miRNAs in patients at risk for metabolic syndrome. Participants included 54 patients from a Family Medicine Clinic at Marshall University School of Medicine and were grouped based on BMI and metabolic syndrome diagnosis: Control, Obese, Metabolic Syndrome (MetS). Serum levels of leptin, adiponectin, leptin: adiponectin ratio, IL-6, six microRNAs (320a, 197-3p, 23-3p, 221-3p, 27a-3p, and 130a-3p), were measured. Among the three groups, leptin, and leptin: adiponectin ratio, and IL-6 levels were highest in MetS, and levels in Obese were greater than Control (p>0.05). Adiponectin levels were lower in Obese compared to Control, but lowest in MetS (p<0.05). MiRNA levels were lowest MetS, and levels in Obese were lower than Control (p>0.05). Our results support the clinical application of biomarkers to diagnose early stage metabolic syndrome, in this population. This would enable attenuation of disease progression before onset of irreversible complications. Since West Virginians are high-risk for developing metabolic syndrome, our biomarker panel could reduce the disease burden on our population.
Abstract P467: Urocortin 2 Levels and Metabolic Profile in Patients With Primary Hypertension
Authors: Walczewska Jolanta , Siga Olga , Dzieza-Grudnik Anna , Wolkow Pawel , Borys Agnieszka , Wizner Barbara , Gryglewska Barbara , Grodzicki Tomasz
Published: July 2021

Objective:
Urocortin 2 (UCN 2) has powerful hemodynamic and neurohormonal actions. Some recent studies suggest that ucn2 may also decrease the steroid production. The aim of this study was to assess the relationship between the serum concentration of UCN 2 and metabolic parameters in patients with primary hypertension.

Methods:
We examined 65 participants with essential hypertension treated with at least 3 antihypertensive medications. The evaluation included anthropometric measurements (BMI, WHR) and ambulatory blood pressure monitoring (ABPM). Moreover we evaluated metabolic parameters in blood serum: glucose, total cholesterol , low-density cholesterol (LDL-C), high density cholesterol (HDL-C), triglycerides (TG), HbA1c, insulin, cortisol, hsCRP. For the analysis of UCN 2 serum level we used ELISA Kit.

We divided the surveyed subjects into two groups according to the urocortin’s median value. The comparative analyses between the groups were based on the student’s T-test, U Mann-Whitney and Chi
2
. To examine the relationships between selected parameters we used Pearson and Spearman correlations.

Conclusion:
Subjects with higher level of UCN 2 have lower concentration of total cholesterol and LDL-C, however changes in UCN 2 concentration were not associated with others metabolic parameters.

Ucn2 may play a protective role in cardiovascular morbidity through reduction of cholesterol synthesis and development of visceral obesity in patients with arterial hypertension.
Abstract P470: Cardiac Autonomic Neuropathy Develops As A Result Of Mild Hypercaloric Intake In Absence Of Signs Of Diabetes: Modulation By Anti-diabetic Drugs.
Authors: El-Yazbi Ahmed , Al-Assi Ola , Ghali Rana , Kaplan Abdullah , Mougharbil Nahed , Eid Ali H , Zouein Fouad A
Published: July 2021

Cardiac autonomic neuropathy (CAN) represents a major cause of morbidity and mortality in diabetes. It is usually seen early in the course of diabetes as an impaired heart rate variability (HRV) and baroreflex sensitivity (BRS), and represents an independent risk predictor of cardiac mortality. CAN development is linked to hyperglycemia; however, current understanding extends cardiovascular risk to pre-diabetic patients with slight glycemic changes. As well, recent evidence suggests that anti-diabetic drugs (metformin and pioglitazone) reduced the risk of cardiovascular complications in pre-diabetic patients. Here, we assessed whether CAN develops independent of hyperglycemia and whether metformin or pioglitazone modify this process. Rats were fed a hypercaloric (HC) diet (4.035 KCal/g vs. 3 KCal/g for control rats) composed of: weight (calories) 18.06 % fat (38.68%), 15.8% protein (15.66%), and 46.13% carbohydrates (45.73%). Stable fasting hyperglycemia developed by 16 weeks of feeding. However, at 12 weeks of feeding, there was no elevation in body weight, fasting or random blood glucose, and no difference in oral glucose tolerance, yet an increase in adipose inflammatory cytokines was observed (4- and 40- fold increase in IL-1β and TGF-β expression). No change in systolic blood pressure was observed over the course of feeding. At 12 weeks, carotid and jugular access were established. Mean arterial pressure (MAP) and heart rate (HR) were recorded, and BRS was assessed using Oxford method. HC-fed rats had a higher pressor response to increasing i.v. doses of phenylephrine vs. control rats. BRS sensitivity was blunted (slope of the ΔMAP vs. ΔHR line, -1.018 ± 0.1217 vs. -0.3379 ± 0.04135) indicating reduced parasympathetic feedback. A 2-week treatment with pioglitazone (2.5 mg/Kg) or metformin (100 mg/Kg) normalized the adipose cytokine profile, yet only pioglitazone improved BRS (-0.7463 ± 0.05775). Parasympathetic dysfunction in HC fed rats was further demonstrated by a decreased high frequency power upon frequency domain analysis of HRV data (3098 ± 233 vs. 89 ± 88 μs
2
). To our knowledge, this is the first report that CAN occurs prior to any glycemic alterations with a potential role for adipose inflammation and modification by antidiabetic drugs.
Abstract P472: Prorenin Independently Causes Hypertension And Renal and Cardiac Fibrosis In Cyp1a1-Prorenin Transgenic Rats
Authors: Huang Yufeng , Zhou Guangyu , Wu Jie , Gu Chunyan , Cheung Alfred
Published: July 2021

Plasma prorenin is commonly elevated in diabetic patients and appears to predict the development of diabetic nephropathy. However, the pathological role of prorenin is unclear. In this study, a transgenic, inducible, hepatic prorenin-overexpressing rat model was generated and the effect of prorenin in organ injury was examined. Four groups of rats (Cyp1a1 prorenin transgenic male and female rats and nontransgenic littermates) were assigned to receive a diet containing 0.3% of the transgene inducer indole-3-carbinol (I3C) for 4 weeks. Plasma prorenin concentration rose from 23±6 to 208±44 (μg/ml) and MAP increased from 77±5 to 138±17 (mmHg), whereas renal prorenin/renin protein expression was unchanged, in transgenic rats fed with I3C diet. The intact prorenin, not renin, in plasma and urine samples was further observed by western blot analysis. Importantly, transgenic rats with high levels of prorenin developed albuminuria, glomerular and tubulointerstitial fibrosis associated with increased expression of TGFß1, PAI-1, collagen and fibronectin. These rats also exhibited cardiac hypertrophy determined by echocardiography, with elevated ratio of heart weight to body weight. Cardiac collagen in interstitial and perivascular area was prominent, accompanied by the increases in mRNA contents of ANP, BNP, ß-MHC, TGFß1 and PAI-1 in the heart tissue. Furthermore, renal protein levels of phosphor-NF-kB-p65 and MCP-1, NAPDH oxidase and MDA, phospho-ß-catenin and phospho-Akt were dramatically increased in prorenin overexpressed rats. These results indicate that prorenin, without being converted to renin, causes arterial hypertension, renal and cardiac fibrosis independently via the induction of inflammation, oxidative stress and the ß-catenin and Akt-mediated signals.
Abstract P473: Group IV Cytosolic Phospholipase A2α is Critical for Norepinephrine-Induced Hypertension
Authors: Pingili Ajeeth K , Song Chi Yong , Shin Ji Soo , Bonventre Joseph V , Malik Kafait U
Published: July 2021

Previously we reported that angiotensin (Ang) II-induced hypertension and associated cardiovascular and renal dysfunction are mediated by cytosolic phospholipase A
2
α (cPLA
2
α) activation, the release of arachidonic acid (AA), and production of eicosanoids predominantly with pro-hypertensive effects (
Hypertension. 2015; 65: 784-792; 2016; 29: 258-265
). We have also shown that norepinephrine (NE) by activating cPLA
2
releases AA, and production of prostanoids in vascular smooth muscle cells (
J Biol Chem. 1996; 217:30149-30157; J. Pharmacol. Exp. Ther. 1993; 266: 1113–1124
).

This study was conducted to determine the contribution of cPLA
2
α in NE-induced hypertension. Eight weeks old male wild-type (cPLA
2
α
+/+
) and cPLA
2
α gene disrupted (cPLA
2
α
-/-
) mice were infused with NE (10 mg/kg/day, s.c.) or its vehicle using mini-osmotic pumps for 2 weeks, and the systolic blood pressure (SBP) was measured by tail-cuff. Infusion of NE increased the SBP in cPLA
2
α
+/+
mice (148±3 vs. 118±3 mmHg, P<0.05, n=4-5); but not in cPLA
2
α

-/-

mice (122±5 mmHg, n=5). The NE-induced increase in SBP was minimized by treatment with AA metabolism inhibitor, 5,8,11,14-eicosatetraynoic acid (ETYA) (25 mg/kg, i.p., every 3
rd
day) in cPLA
2
α
+/+
mice (125±5 vs. 148±3 mmHg, P<0.05, n=4-5). Prostaglandin (PG) E2-EP1 and EP3 receptor activation that increase blood pressure have been implicated in Ang II-induced hypertension. In our study antagonists of the EP3 receptor (L-798106) (10 mg/kg, i.p. every 3
rd
day) decreased the NE-induced increase in SBP (130±5 vs. 148±3 mmHg, P<0.05, n=5/group). These data suggest that cPLA
2
α contributes to NE-induced increase in SBP via cPLA
2
α activation, the release of AA and generation of eicosanoids, most likely PGE2 that exerts pro-hypertensive effects by stimulating EP3 receptors. Therefore, the development of agents that selectively inhibit the cPLA
2
α activity or block EP3 receptors could be useful in treating hypertension and its pathogenesis.
Abstract P480: Effects Of Long And Intermediate Acting Dihydropyridine Calcium Channel Blockers In Hypertension: A Systematic Review And Aeta-analysis Of 18 Prospective, Actively Controlled, Randomized Clinical Trials
Authors: Chaugai Sandip , Arima Hisatomi , Sherpa Lhamo Yangchen , Sepehry Amir
Published: July 2021

Background:
Dihydropyridine calcium channel blockers are a heterogeneous group of antihypertensive drugs. Long acting dihydropyridine agent Amlodipine is widely used for mono and combination therapy for hypertension in clinical practice, while intermediate acting dihydropyridine agents such as Felodipine, Nifedipine-GITS etc. have shown inconsistent results in randomized clinical trials (RCTs).

Methods and Results:
A meta-analysis of 18 prospective, actively controlled RCTs enrolling a total of 80,069 hypertensive patients followed for a mean of 51.7 months was performed. Amlodipine therapy was associated with 24% higher risk of heart failure (RR: 1.24, [95%CI: 1.04 to 1.48], p= 0.019) but 16% lower risk of stroke (RR: 0.84, [95%CI: 0.73 to 0.95], p=0.009) without statistically significant effect on acute myocardial infarction (AMI) (RR: 0.90, [95%CI: 0.78 to 1.03], p=0.128) or cardiovascular event (CVE) (RR: 0.99, [95%CI: 0.93 to 1.06], p=0.814). Intermediate acting dihydropyridine calcium channel blocker therapy was associated with 24% higher risk of heart failure (RR: 1.24, [95%CI: 1.07 to 1.44], p=0.005), 27% higher risk of AMI (RR: 1.27, [95%CI: 1.04 to 1.55], p=0.019), 14% higher risk of CVE (RR: 1.14, [95%CI: 1.01 to 1.29], p=0.034) without statistically significant effect against stroke (RR: 1.03, [95%CI: 0.87 to 1.21], p=0.580). Amlodipine showed higher risk of heart failure (RR: 1.20, [95%CI: 1.00 to 1.43], p=0.045) and lower risk of stroke (RR; 0.85, [95%CI: 0.75 to 0.96], p= 0.007) compared to Renin-angiotensin-system (RAS) blockers. Intermediate acting dihydropyridine calcium channel blocker showed higher risk of heart failure (RR: 1.31, [95%CI: 1.09 to 1.57], p=0.004) and AMI (RR: 1.46, [95%CI: 1.00 to 2.14], p=0.050) compared to RAS blockers. Multivariate meta-regression suggested that the effect size may differ between intermediate and long acting agents for stroke (B: -0.27, [95%CI: -0.50 to -0.05], p=0.012) and AMI (B: -0.42, [95%CI: -0.69 to -0.16], p=0.002) after adjustment for baseline presence of diabetes.

Conclusions:
This study suggests that Amlodipine offers greater protection against major complications of hypertension compared to intermediate acting dihydropyridine calcium channel blockers.
Abstract P482: Different Hemodynamic Phenotypes Of Isolated Systolic Hypertension In Adolescents
Authors: Obrycki Lukasz , Niemirska Anna , Sarnecki Jedrzej , Kulaga Zbigniew , Litwin Mieczyslaw
Published: July 2021

Objective:
Isolated systolic hypertension (ISH) is a dominant form of primary hypertension (PH) in adolescents. Some of them present with normal central systolic blood pressure (cSBP), a phenomenon called spurious hypertension (sHT). The study was aimed to describe hemodynamics of PH in relation to cSBP, central pulse pressure (cPP) and target organ damage (TOD) in adolescents referred because of PH.

Patients and Methods:
In 267 children (59 girls; 14.9 ±2.6 years) referred with arterial hypertension, in whom secondary hypertension was excluded, 24 hour ABPM, left ventricular mass index (LVMi), carotid intima-media thickness (cIMT), pulse wave velocity (PWV), cSBP, cPP, cardiac index (CI) and stroke volume (SV) was assessed. 64 age and sex matched normotensive control children were control group.

Results:
145 subjects had white coat hypertension (WCH) including 24 with ambulatory prehypertension (ambpreHT). Of 122 hypertensive pts, 39 had ambulatory hypertension (ambHT) and 83 severe ambulatory hypertension (severeHT). Normal cSBP was found in all WCH subject and 23 with ambpreHT. 39 of 122 (32%) hypertensive pts had sHT - 47.4% in those with ambHT and 26.5% with severeHT (p=0.0001). cIMT, LVMi, PWV, cSBP and cPP increased across blood pressure strata from normotension, through sHT to PH with elevated cSBP (all p<0.05). LVMi and cIMT correlated with cSBP (r = 0.220; p = 0.0007; r = 0.14; p = 0.04, respectively) and cPP (r = 0.274; p = 0.0001; r=0.202; p=0.002, respectively). 36 pts with left ventricular hypertrophy (LVH) had greater cPP (52 ±10 mmHg) in comparison with subjects without LVH (47 ±8 mmHg; p = 0.027). Regression analysis revealed cPP as the only predictor of LVMi (r
2
= 0.09, β = 0.143, p = 0.03). ROC area for predictors of LVH revealed similar area under curve for cSBP (0.585), cPP (0.618) and 24h systolic ABPM (0.612). Patients with sHT had greater amplification of pulse pressure than normotensive ones. CI and SV was lowest in normotensive controls, intermediate in sHT patients and highest in patients with elevated cSBP (p<0.05).

Conclusions:
sHT present with intermediate hemodynamic phenotype between normotension and sustained PH. cSBP and cPP differentiates patients with severeHT and TOD from patients with WCH, ambpreHT and ambHT without TOD
Abstract P483: Healing a Broken Heart Using a Closed-loop, Allostatic, Neurotechnology: A Case Study in a Patient Suffering From Takotsubo Syndrome
Authors: Tegeler Charles H , Tegeler Catherine L , Shaltout Hossam A
Published: July 2021

Background:
Stress cardiomyopathy or Takotsubo syndrome (TS) is an acute, reversible disorder of the heart characterized by left ventricular dysfunction, usually triggered by a stressful event. Excessive sympathetic excitation and shift in symapthovagal balance are proposed as mechanisms. High-resolution, relational, resonance-based, electroencephalic mirroring (HIRREM®) is a noninvasive, closed-loop, allostatic, neurotechnology using software algorithms to identify specific brain frequencies, translating them in real time into audible tones, to support self-optimization of brain rhythms and improve autonomic balance.

Objective:
To evaluate benefits of HIRREM on autonomic function and symptom reduction in a 55 year old female enrolled in an IRB-approved open label feasibility study, after TS diagnosis.

Results:
The participant received 13 HIRREM sessions over 11 days (9 in office days). Data were collected before, and 21 days after HIRREM completion. Baseline brain patterns had prominent right dominance at temporal lobes (sympathetic pattern), which balanced by the end of the sessions. Cardiovascular autonomic balance also shifted away from sympathetic towards parasympathetic. This was seen as reduced LF/HF ratio (from 1.89 to 0.63), increased heart rate variability (rMSSD from 27 to 40.8 ms), and baroreflex sensitivity (from 11.8 to 24.4 ms/mmHg). Blood pressure dropped from 132/90 to 121/88 with no change in heart rate despite discontinuation of her ACE inhibitor medication due to her BP being “too low.” HIRREM use was also associated with clinically meaningful improvements in multiple symptom inventories including insomnia (ISI) from 15 to 6, depression (CES-D) from 16 to 2, anxiety (GAD-7) from 18 to 2, and perceived stress scale (PSS) from 30 to 14. The patient reported resolution of a feeling of heaviness and discomfort in the chest after starting HIRREM sessions.

Conclusion:
These data provide the first report of potential cardiovascular benefits of a non- pharmacological therapy to patients suffering from broken heart syndrome.
Abstract P486: Effects of Isometric Leg Training on Ambulatory Blood Pressure and Morning Blood Pressure Surge in Young Normotensive Men and Women
Authors: Baross Anthony W , Milne Kevin J , McGowan Cheri L , Swaine Ian L
Published: July 2021

Introduction:
Cardiovascular disease (CVD) is a major cause of death globally with hypertension reported to be a leading modifiable risk factor. Ambulatory blood pressure (BP), in particular diurnal BP variability, is considered to be associated with CVD risk. In addition, the morning BP surge (MBPS) is thought to be associated with increased stroke risk and to be a destabilizing factor for atherosclerotic plaque. Isometric resistance training (IRT) is an effective method of lowering BP and has been recommended by the American Heart Association as an alternative treatment for reducing BP. To date, few studies have investigated the effects of IRT on ambulatory BP and particularly the morning surge in BP. Therefore, the purpose of this study was to determine whether (i) IRT causes reductions in ambulatory BP and the MBPS, in young normotensives and (ii) there are any sex differences in these changes.

Methods:
Ambulatory BP was measured prior to, and after, 10 weeks of bilateral leg IRT using an isokinetic dynamometer (4 x 2 minute contractions at 20% MVC, with 2 minute rest periods on 3 days per week). Twenty normotensive individuals (10 men, age=21 ± 4 years; 10 women, age=23 ± 5 years) were recruited. A two-way repeated measures ANOVA was used to assess the within and between groups ambulatory (mean 24-h, daytime, night time and diurnal variation) BP and MBPS. MBPS was calculated as: mean systolic BP 2 hours after waking minus the lowest sleeping 1 hour mean systolic BP.

Results:
There were significant reductions in 24-h ambulatory (4 ± 2 mmHg, p=0.0001; 4 ± 2 mmHg, p=0.0001) systolic BP in both men and women following IRT. This comprised significant reductions in day time (5 ± 5 mmHg, p=0.019; 5 ± 4 mmHg, p=0.002) but not night time (1 ± 5 mmHg, p=0.75; 1 ± 3 mmHg, p=0.3) systolic BP. Additionally, there were significant reductions in the MBPS (6 ± 8 mmHg, p=0.044; 6 ± 7 mmHg, p=0.019). There were no significant differences between men and women in these changes (p>0.05).

Conclusion:
These results support previous research showing that IRT is effective in lowering ambulatory BP. Furthermore, the significant reductions in the MBPS offer the potential for clinically meaningful CVD and stroke risk reduction, provided these effects can be demonstrated in those who are at risk.
Abstract P496: Effects of
Nitrosomonas Eutropha
D23 Topical Spray on Blood Pressure: Results From a Randomized, Double-blind, Vehicle Controlled, Dose-ranging Study in Normotensive Adults
Authors: Hino Peter , Liu Yan , Jiang Lilly , Weiss Lawrence , Jamas Spiros , Neutel Joel M
Published: July 2021

Objective:
To evaluate the safety and blood pressure effects in normotensive individuals of
Nitrosomonas eutropha
bacteria applied as a topical spray.

Methods:
After initial screening, 36 healthy, normotensive adults participating in a facial acne vulgaris treatment trial were randomized to placebo or one of three concentrations (2, 4, or 8 x10
9
cells/ml) of a pure culture of ammonia oxidizing bacteria strain
N. eutropha
D23. Treatment occurred through the application of a topical spray to the face, which delivered ~0.6ml twice daily for 14 days. A standardized safety assessment (including physical exam and 12-lead EKG) and an evaluation of systolic and diastolic blood pressure occurred on Days 1 (pre-treatment), 3, 7, 10, 15 and on Day 28 following a 14 day wash-out.

Results:
A total of 9 subjects in each treatment group were randomized and completed the study (8 men and 28 women). Pre-treatment blood pressure was similar across treatment groups (115-119/75-79 mmHg). A dose-dependent reduction in systolic and diastolic blood pressure was emergent by treatment day 3. At day 15, individuals treated with the 8 x10
9
cells/ml concentration experienced a reduction in systolic blood pressure of 6.1 mmHg from baseline (SD=5.9; p=0.014) and diastolic blood pressure was reduced by 3.56 mmHg from baseline (SD=4.49; p=0.041). By comparison, in the placebo group, the treatment phase systolic and diastolic blood pressure changes from baseline were -0.34 (SD=7.80; p=0.94) and -1.19 (SD=4.57; p=0.486) respectively. Blood pressure reductions observed at Day 15 with
N. eutropha
were generally maintained at Day 28. All 3 concentrations of
N. eutropha
and placebo (vehicle alone) were well tolerated and no safety issues were identified.

Conclusions:
Topical treatment with the ammonia oxidizing bacteria
N. eutropha D23
produced a dose-dependent reduction in systolic and diastolic blood pressure that was statistically significant in the 8 x10
9
cells/ml treatment group. This effect is consistent with the proposed mechanism-of-action of
N. eutropha
treatment: modification of NO/NO
2
levels on the skin. These findings support further development of
N. eutropha
for treatment of acne vulgaris and evaluation of blood pressure lowering effects in hypertensive individuals.
Abstract P499: Protein Kinase Cα Deletion Causes Hypotension Due to Decreased Vascular Contractility
Authors: Wynne Brandi M , McCarthy Cameron G , Szasz Theodora , Klein Janet D , Webb R. Clinton , Hoover Robert S
Published: July 2021

Protein kinase Cα (PKCα) regulates multiple cell signaling pathways, including those that impact blood pressure. PKCα activation increases vascular smooth muscle contractility, yet reduces cardiac contractility. PKCα has also been shown to modulate nephron ion transport. We have shown that PKCα deletion leads to hypotension, with compensatory increases in sodium retention. Here, we hypothesized that PKCα deficiency reduces vascular contractility, leading to decreased mean arterial pressure (MAP). MAP, measured by telemetry, was decreased in PKC KO (≈12 mmHg) compared to PKC control (PKC CTL) mice. Aorta and mesenteric arteries were isolated, and concentration response curves (CRCs) to phenylephrine (Phe), acetylcholine (ACh) or sodium nitroprusside (SNP) were performed in the presence of vehicle or the following inhibitors: L-NAME or indomethacin (NOS, COX inhibitor,
resp.
). CRCs to KCL were performed to assess receptor-independent vascular responses. In aorta, we observed a striking reduction in KCl-mediated contraction (5.8±0.3mN
vs.
10.4±1.1mN control, **p<0.01). PKC KO aorta and mesenteric arteries had decreased contractile responses to Phe, as compared to control (aorta, 12.7±0.5mN

R
max

vs.
16.3±0.5mN

R
max

, and mesenteric 9.9±0.3mN

R
max

vs.
11.8±0.6mN

R
max

; n=4, **p<0.01), revealing a role for reduced vascular contractility. Endothelium-mediated relaxation responses to ACh were also increased in PKC KO mice, as compared to control (59.3±6.8%

R
max

vs.
45.4±3.2%

R
max

, n=4, *p<0.05). Interestingly, NOS inhibition increased contractility in mesenteric arteries from PKC KO mice (8.55±2.65mN

R
max

vs.
6.95±0.39mN

R
max

control, n=4, ***p<0.001). However, PKC KO aorta had an enhanced response to COX inhibition (12.2±0.7mN

R
max

vs.
10.1±0.6mN

R
max

control, n=4, *p<0.05) suggesting that PKCα may be negatively regulating NOS in mesenteric arteries, and COX-mediated prostaglandin production in the aorta. No differences were observed in the relaxation responses to SNP. These data suggest that global deletion of PKCα results in hypotension due to decreased vascular contractility, and loss of PKCα-mediated inhibition of endothelial relaxing factors. Thus, systemic targeting of PKCα may be beneficial for the reduction of MAP.
Abstract P500: Activation of the Heterodimeric Erythropoietin /β-Common Receptor Impairs Acetylcholine Mediated Vasodilation in Mouse Mesenteric Arterioles
Authors: Kilar Cody R , Diao YanPeng , Sautina Larysa , Sekharan Sivakumar , Keinan Shahar , Conrad Kirk P , Segal Mark S , Mohandas Rajesh
Published: July 2021

Erythropoietin (EPO) increases systemic vascular resistance and blood pressure. However, endothelial cells cultured in the presence of EPO demonstrate increased production of the potent vasodilator, nitric oxide (NO). The mechanism by which EPO causes vasoconstriction despite stimulating NO production may be dependent on its ability to differentially activate the two receptor complexes, the homodimeric EPO (EPOR
2
) and the heterodimeric EPOR/β-common receptor (βCR).

Objective:
The purpose of this study was to investigate the contribution of the EPOR
2
and βCR receptor to the vasoactive properties of EPO.

Methods:
First order, mesenteric arteries isolated from 16-week old male C57BL/6 mice were cannulated and perfused using a pressure arteriography system. To determine the contribution of each receptor complex, arteries were incubated with EPO stimulating peptide (ESP) which binds and activates only the heterodimeric EPOR/βCR complex or EPO which activates both receptors, 20 min prior to evaluation of vasoconstrictor (phenylephrine and potassium chloride), endothelium-dependent (acetylcholine, bradykinin, A23187) and -independent (sodium nitroprusside) vasodilator responses. Additionally, we studied the effect of a novel βCR inhibitory peptide (βIP) which was developed in silico and validated by demonstrating that it selectively inhibits binding of ligands to the βCR.

Results:
Acetylcholine induced vasodilation was impaired in arteries pretreated with EPO or ESP by 100% and 60%, respectively. EPO and ESP did not affect endothelium-dependent vasodilation by Bradykinin or A23187, endothelium-independent vasodilation by sodium nitroprusside, or vasoconstriction by phenylephrine and KCl. The βIP prevented the impairment of acetylcholine-induced vasodilation by EPO and ESP.

Conclusion:
Together, our findings suggest that activation of the heterodimeric EPOR/βCR leads to selective impairment of ACh-mediated vasodilator response in mouse mesenteric resistance arteries. Thus the βCR might have a role in mediating hypertensive effects of EPO. Therapeutic inhibition of the βCR might prevent vascular complications of EPO without affecting erythropoiesis.
Abstract P502: Vascular Tissue Proteomics Enables Detection of Cell Type-specific Changes in Target Expression
Authors: Mackenzie Ruth M , Pinel Karine , Carrick Emma J , Nather Katrin , Husi Holger , Graham Delyth , Mullen William , Nicklin Stuart A , Delles Christian
Published: July 2021

Introduction:
Identification of robust targets from proteomic studies in cardiovascular tissue may prove challenging. Using an angiotensin II (Ang II)-infusion mouse model, we performed a proteomics study in isolated thoracic aortas. Changes in proteins related to cardiovascular pathophysiology were identified and candidate targets selected for validation via traditional techniques.

Methods and Results:
C57 black/6 mice were infused with Ang II (24 μg/kg/hour) via osmotic minipump for 6 weeks. Elastic Van Gieson staining demonstrated significantly increased medial area in aortas from Ang II-infused mice compared to water-infused control mice (
P
<0.005 ), indicating Ang II-induced remodelling.

Nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS) revealed a 1.28 fold increase in galectin-3 (LGALS3) expression in vessels from Ang II-infused mice as compared to controls. LGALS3, a β-galactoside binding lectin, is a well known marker of cardiovascular disease reported to play a role in Ang II-induced cardiac remodelling.

Immunohistochemical (IHC) staining showed increased LGALS3 expression throughout the vessel wall and particularly in the endothelial layer (quantification using Image J Fiji software: 110.4 ± 1.37 vs 120.5 ± 2.6 arbitrary units;
P
=0.005) in Ang II-infused mice compared to controls.

Human primary endothelial cells (ECs) were isolated from saphenous veins of patients undergoing coronary artery bypass graft surgery and translational studies performed.
LGALS3
/LGALS3 expression was detected at both mRNA and protein level by qRT-PCR and immunoblotting respectively. Acute stimulation of ECs with Ang II (200nM for 24 hours) failed to upregulate
LGALS3
/LGALS3 expression suggesting that the increased endothelial expression observed
in vivo
is due to chronic infusion of Ang II.

Conclusions:
We have successfully validated the Ang II-induced increase in LGALS3 identified via vascular tissue proteomics. Despite the use of homogenised whole aortic tissue, nano LC-MS/MS proved sensitive enough to detect elevated expression of a candidate protein that is predominantly expressed in the endothelium. Tissue proteomics can detect changes in expression specific to a single cell type.
Abstract P517: Dialysis Vintage Longer Than Sixty Months Contributes to Increased Arterial Stiffness and Impaired Diastolic Function in Patients with End-stage Renal Disease
Authors: Calvez Valentin , Palladino Marco , Montefusco Giulio , Cesario Vincenzo , Fofi Claudia , Ciavarella Giuseppino M , Volpe Massimo , Savoia Carmine
Published: July 2021

Hemodialysis (HD) may induce vascular stiffness through several mechanisms. We sought to determine the role of dialysis vintage (DV) on the development of cardiovascular alterations. We studied 14 patients in chronic HD and 24 newly diagnosed never treated hypertensive patients and 16 normotensive controls.

The patients in HD were divided in two groups according to DV: <60-months (DV<60,n=7) or >60-months (DV>60,n=7). After HD session, when dry weight was reached, we evaluated peripheral blood pressure (pBP), the parameters derived by tonometric analysis of the pulse waveform (central blood pressure-cBP-, Subendocardial Viability Ratio-SEVR-, carotid-femoral pulse wave velocity-cf-PWV-) and those derived from echocardiography: ejection fraction (EF-for systolic function) and E/e’ (for diastolic function), and the ultrafiltration volume (UV). Calcium/phosphate (Ca/P) levels, serum albumin, and Kt/V were evaluated retrospectively on repeated measurements over the past 5 years.

All the groups were similar for sex and BMI, both DV<60 and DV>60 were older than hypertensives and controls (58.33±3.71 and 59.83±7.98 vs 44.14±1.28 and vs 40.63±2.05 years, respectively, P<0.05). Both DV<60 and DV>60 presented similar levels of Ca/P, serum albumin, Kt/V and UV. pBP was increased and similar to hypertensives in DV>60 vs DV<60 (systolic-pBP: 154.2±4.5 vs 132.5±5.18 mmHg, P<0.01 and diastolic-pBP: 90.4±49 vs 78.5±3.3 mmHg, P<0.01). Likewise cBP was increased and similar to hypertensive patients in DV>60 vs DV<60 (systolic-cBP: 140.8±8.4 vs 111.2±3.36 mmHg, P<0.001 and diastolic-cBP: 88.2±3.73 vs 72.33±7.78 mmHg, respectively, P<0.05). cf-PWV was similar in normotensives, hypertensives and DV<60, and increased only in DV>60 vs DV<60 (9.6±1.4 vs 7.13±1.4 m/s, p<0.05). SEVR and EF were preserved and similar in all the groups. E/e’ was significantly increased only in the groups in HD, however it was higher in DV>60 vs DV<60 (9.16±1.14 vs 6.96±0.72, P<0.01).

In conclusion, only patients with DV>60 presented increased aortic stiffness. This was associated to higher BP and diastolic dysfunction. Hence, chronic HD, particularly after 60 months, may play a putative role in developing cardiovascular alterations in patients with end-stage renal disease.
Abstract P529: Renal
Authors: Konkalmatt Prasad , Han Fei , Xu Gaosi , Zheng Xiaoxu , Gildea John J , Felder Robin A , Jose Pedro A , Armando Ines
Published: July 2021

Some common single nucleotide polymorphisms (rs6276 and rs6277, SNPs) of the human
DRD2
gene are associated with decreased D2R expression and function and increased blood pressure or hypertension. Human renal proximal tubule cells (hRPTCs) from subjects carrying these SNPs (hRPTCs-SNPs) express elevated levels of proinflammatory and profibrotic proteins, indicating that the D2R has protective effects in these cells and that decreased D2R function may contribute to the susceptibility to renal disease associated with essential hypertension. Micro RNA 4301 (miR4301) is an intronic miRNA that resides in the second intron of the primary human
DRD2
transcript. A mouse homolog of miR4301 has not been identified to date. We hypothesized that miR4301 expression and function are decreased in hRPTCs-SNPs and that loss of miR4301 mediates, in part, the deleterious effects of decreased D2R function. We studied four cell lines carrying no SNPs (hRPTCs-WT) and four hRPTCs-SNPs lines. miR4301 expression was lower in hRPTCs-SNPs than in hRPTCs-WT (0.52±0.07- vs 1.03±0.14-fold; P<0.05). Silencing D2R via siRNA in hRPTCs-WT also decreased miR4301 expression (0.59±0.12- vs 1.01±0.11-fold; P<0.05). We measured the expression of several genes with pro-inflammatory and pro-fibrotic effects identified by Target 7.1 as miR4301 targets. The mRNA expressions of
SMAD1
(2.7-fold);
CASP2
(4.13-fold),
CASP7
(3.0-fold),
TGFβR1
(4.9-fold), and
CTGF
(7.3-fold) were increased in hRPTCs-SNPs, in comparison with hRPTCs-WT. Moreover, the mRNA expression of the miR4301 target
LEF1
in hRPTCs-WT transfected with miR4301 mimic was lower (0.70±0.02 vs 1.0±0.05-fold; P<0.05) than in cells transfected with control miR, while transfection with miR4301 inhibitor increased the expression of
LEF1
mRNA (1.30±0.03 vs 1.0±0.05-fold; P<0.05), compared with the control miR indicating that miR4301 represses
LEF1
expression. These results show that miR4301 mediates, in part, the protective effects of D2R expression and function on renal injury by repressing the expression of genes related to fibrosis and inflammation and suggest that the function of D2R-related proteins may be dependent, as well, on the regulation of specific miR4302.
Abstract P531:
FOXO3
, a Cardiovascular Protector, is at the Hub of a 46-gene Cell Resilience “Gene Factory” on Human Chromosome 6
Authors: Morris Brian J , Donlon Timothy A , Chen Randi , Masaki Kamal H , Allsopp Richard C , Willcox D. Craig , Elliott Ayako , Willcox Bradley J
Published: July 2021

The transcription factor FoxO3 regulates multiple genes involved in cell resilience. We have previously implicated variation in non-coding DNA of the FoxO3 gene (
FOXO3
) with lower blood pressure, reduced inflammation, less hypertension, reduced coronary heart disease mortality, and longevity. The aim of the present study was to determine transcriptional, genetic and genomic mechanisms involving
FOXO3
. By DNA sequencing of chromosome 6q21 in lymphoblastoid cell lines of 95 men who had survived to ≥ 95 years of age we identified 110
FOXO3
single nucleotide polymorphisms (SNPs). Thirteen SNPs were at binding sites for 18 transcription factors. Those SNPs appeared to be in physical contact, via RNA polymerase II binding chromatin looping, with sites in the
FOXO3
promoter, and likely function together as a
cis
-regulatory unit. At the chromosome level,
FOXO3
was located at the center of a 7.3 Mb 46-gene chromatin domain flanked by gene deserts. We identified distant contact points between
FOXO3
and these 46 neighboring genes, through long-range physical contacts via CCCTC-binding factor zinc finger protein (CTCF) binding sites. The genes in this “archipelago” of neighbourhood genes mediate a similar repertoire of functions as FoxO3, including stress resistance, nutrient sensing, cell proliferation, autophagy, apoptosis and stem cell maintenance. The 7.3 Mb gene domain was highly conserved across species, indicating evolutionary importance. We believe that
FOXO3
serves as the hub for an “interactome” involved in healthy aging, including cardiovascular disease reduction, in those with favorable
FOXO3
genotypes. In support, we found that cellular stress (H
2
O
2
) could stimulate
FOXO3
expression in 20 lymphoblastoid cell lines, being 3-fold stronger for those with a favorable
FOXO3
genotype. In FISH experiments, stress-induced activation of
FOXO3
caused it to move towards its neighboring genes as suggested by our genomic data. In conclusion, we have shown, for the first time, that
FOXO3
is at the central hub of a gene network on chromosome 6 involved in cell protection and healthy aging. The concept of “gene factories” may apply more broadly to genome and genetic mechanisms involved in cardiovascular disease etiology.
Abstract P541: Ambulatory Blood Pressure and Endothelial Dysfunction an Hematopoietic Cell Transplant Patients
Authors: Pao Emily , Gove Nancy , Flynn Joseph , Hingorani Sangeeta
Published: July 2021

Hematopoietic cell transplantation (HCT) is a common treatment for malignancies, metabolic, and genetic disorders. Albuminuria post-HCT, which may represent systemic endothelial injury or inflammation from graft vs host disease, increases the risk of kidney disease and non-relapse mortality at 1 year post-HCT. HCT patients are also known to have abnormal blood pressure (BP) and increased rates of cardiovascular complications. We sought to determine the relationships between albuminuria, endothelial dysfunction and ambulatory BP in HCT patients.

Patients ≥ 12 years of age who had their first allogeneic HCT between 2012-2015 and survived through day 80 were eligible. Peripheral endothelial function was assessed using the EndoPAT2000 device at day 80 post HCT along with a 24 hour ambulatory blood pressure monitor (ABPM) study. Clinical and lab data were collected and a urine sample for an albumin to creatinine ratio (ACR). Both logistic and linear regression analyses were used to identify associations between EndoPAT score and clinical variables including eGFR, 24 hour ABPM study, albuminuria, and serum lipids.

Sixty patients completed the study. The median age was 48 years (range, 14-69). The median EndoPAT score (RHI normal ≥ 1.69) was 2.05 (range, 1.02- 4.45) and 28% (17/60) of the patients had abnormal endothelial function. Forty-two patients (72%) had hypertension based on the 24-hr ABPM results and 63% of the patients (38/60) had abnormal nocturnal dipping. Hypertension on ABPM (p = 0.04), abnormal nocturnal dipping (p=0.04), and elevated serum triglyceride levels (p=0.03) were associated with a lower EndoPAT score. Albuminuria was not associated with EndoPAT score or hypertension based on ABPM. Abnormal nocturnal dipping was associated with lower eGFR (p=0.03). There was a lack of agreement between office BP and use of medications and results on a 24 hour ABPM study (p < 0.001).

We did not find a correlation with lower EndoPAT scores and albuminuria; we did find associations between abnormal nocturnal dip, elevated triglyceride levels and hypertension diagnosed by 24 hour ABPM. Casual office BP readings do not accurately reflect the HCT patient’s true BP suggesting that 24 hour ABPM studies are needed to diagnose and treat hypertension appropriately.
Abstract P542: Coenzyme Q10 Supplementation in Orthostatic Hypotension and Multiple-system Atrophy, Report on 5 Cases
Authors: Rembold Christopher
Published: July 2021

Multiple-system atrophy is a neurologic disorder characterized by orthostatic hypotension, Parkinsonian signs, and cerebellar signs. Mutations in COQ2, an enzyme involved in coenzyme Q10 (CoQ10) synthesis were recently associated with familial and sporadic cases of multiple-system atrophy. I hypothesized that people with symptoms suggesting multiple-system atrophy might benefit from CoQ10 administration. Five patients with symptomatic orthostatic hypotension were treated in an unrandomized manner with 240±40 mg CoQ10 daily for 8±3 months - two had Parkinsonian and one had cerebellar signs. Prior to starting CoQ10, systolic BP fell 30±5 mmHg upon standing from a sitting position. After treatment with CoQ10, systolic BP fell 8±5 mmHg upon standing from a sitting position (p=0.013 for change in systolic BP fall by paired t test). These data suggest that orthostatic hypotension could possibly improve with CoQ10 administration and that a randomized clinical trial to test this hypothesis should be begun.
Abstract P544: Osteopontin Deficiency Decreases Autophagy and Exacerbates Tubular Injury in Acute Kidney Injury Induced by Folic Acid
Authors: Nagao Tomoaki , Okura Takafumi , Tanino Akiko , Miyoshi Ken-ichi , Kukida Masayoshi , Higaki Jitsuo
Published: July 2021

Osteopontin (OPN), a secreted glycosylated phosphoprotein and pro-inflammatory cytokine, has been implicated in the pathology of several renal conditions, especially renal fibrosis in chronic kidney disease. OPN is slightly expressed in renal tubular cells in normal condition, but after acute tubular injury, OPN is highly induced in these cells. However, the role of induced OPN is still unclear.

The aim of this study was to clarify the roles of OPN in acute kidney injury (AKI).

AKI was induced in wild type (WT) and OPN knockout (KO) mice by using folic acid (FA) injection (0.35mg/kg). After 2days of injection, 34% of WT mice died, whereas 54% of KO died from renal failure. Kidneys from survived mice were removed and the renal histological changes, protein expression were examined. BUN and Creatinine levels were markedly elevated in WT-AKI and KO-AKI mice (BUN: WT-sham; 25.7±4.7mg/dl, WT-AKI; 315.0±173.2mg/dl, KO-AKI; 337.7±163.7mg/dl, Creatinine: WT-sham; 0.08±0.03 mg/dl, WT-AKI; 1.60±0.87 mg/dl, KO-AKI; 1.80±0.94 mg/dl). Renal OPN mRNA expression was increased in WT-AKI mice compared to WT-sham mice (p<0.05). High levels of OPN expression in renal tubular cells were induced in WT-AKI mice. TUNEL positive tubular cells were increased in KO-AKI mice compared to WT-AKI mice. In immunohistochemical analysis, Kidney injury molecules 1 (Kim-1) positive tubular cells were also highly

increased in KO-AKI mice compared to WT-AKI mice. In contrast, LC3B (autophagy related protein) positive tubular cells were decreased in KO-AKI mice compared to WT-AKI mice.

These results indicate that OPN deficiency exacerbates tubular injury via through the inhibiting autophagy in folic acid induced AKI mice.