bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P162:
In vivo
miR-431 Inhibition Protects Against Vascular Damage and Hypertension
Authors: Huo Ku-Geng , Fraulob-Aquino Julio C. , Barhoumi Tlili , Richer Chantal , Coelho Suellen C. , Ouerd Sofiane , Caillon Antoine , Lajoie Mathieu , Sinnett Daniel , Paradis Pierre , Schiffrin Ernesto L.
Published: July 2021

Background:
Vascular injury is an early manifestation of hypertension. microRNAs (miRNAs) play an important role in cardiovascular disease, but their implication in vascular injury remains unclear. Using small and total RNA sequencing, we identified in murine mesenteric arteries (MAs) a conserved angiotensin (Ang) II-upregulated Dlk1-Dio3 miRNA miR-431 that correlated with blood pressure (BP), and an Ang II-downregulated BP-correlated conserved putative miR-431 target, the transcriptional factor ETS homologous factor (
Ehf
). miR-431 might be involved in vascular remodeling as
Ehf
regulates expression of extracellular matrix genes including alpha-1 type I collagen (
Col1a1
) and of other Dlk1-Dio3 miRNAs. In this study, we proposed to validate the miR-431-
Ehf
-
Col1a1
interaction
in vitro
and
in vivo
, and determine whether miR-431 inhibition antagonizes angiotensin (Ang) II-induced hypertension and vascular injury.

Methods and Results:
Transfection of miR-431 mimics into human aortic smooth muscle cells decreased
Ehf
expression (0.13±0.05 fold,
P
<0.001) and increased
Col1a1
(1.7±0.5 fold,
P
<0.01), whereas miR-431 inhibitors increased
Ehf
(1.5±0.2 fold,
P
<0.001) and decreased
Col1a1
(0.89±0.11 fold,
P
<0.05).
Ehf
siRNA transfection increased 1.2±0.1 fold
Col1a1
(
P
<0.01). Co-transfection of miR-431 mimics with luciferase reporter vectors that contain the wild-type but not mutated miR-431 human
EHF
3’ UTR binding site decreased 0.51±0.01 fold (
P
<0.05) luciferase expression compared to scrambled mimics. miR-431 inhibitor IV injection in mice at day 0 and 7 of Ang II infusion decreased miR-431 (0.16±0.05 fold,
P
<0.01),
Col1a1
(0.58±0.11 fold,
P
<0.05), increased
Ehf
(2.9±0.8 fold,
P
<0.05) in MAs, delayed BP elevation (
P
<0.01), improved endothelium-dependent relaxation (33±8 vs 64±7%,
P
<0.05) and reduced vascular stiffness (strain at 140mmHg: 0.68±0.02 vs 0.58±0.02 ΔD/D,
P
<0.01) compared to scrambled mimics-injected Ang II-infused mice.

Conclusion and Perspectives:
miR-431 and its target
Ehf
may act as master regulators in the pathophysiology of vascular damage in hypertension. miR-431 inhibition has the potential to serve as a novel therapeutic approach for treatment of vascular damage and hypertension.
Abstract P187: Nitric Oxide Function in Postural Tachycardia Syndrome During High and Low Sodium Diets
Authors: Celedonio Jorge E , Nwazue Victor C , Garland Emily M , Shibao Cyndya A , Okamoto Luis E , Biaggioni Italo , Raj Satish R , Gamboa Alfredo
Published: July 2021

Background:
Postural tachycardia syndrome (POTS) is characterized by an increase in sympathetic activity, with an exaggerated rise in heart rate upon standing and symptoms of cerebral hypoperfusion. Endothelial/Nitric Oxide (NO) dysfunction might be involved in POTS pathophysiology. As part of the non-pharmacologic treatment of POTS, a high sodium diet is often recommended to increase plasma volume. We assessed endothelial/NO function in conduit vessels (using flow-mediated dilation, FMD) and resistance vessels (using finger pulse arterial tonometry, PAT) in POTS patients during high and low salt diets.

Methods:
In 14 female POTS patients (34±9 years, BMI 23±3 kg/m
2
) and 13 matched healthy control subjects (29±4 years, BMI 24±3 kg/m
2
), we evaluated the time course responses to FMD and PAT. Subjects were randomly assigned to either high salt diet (300mEq/day) or low salt diet (10mEq/day) for 6 days and crossed over to the other arm after 1 month. The areas under the curve for brachial artery diameter by FMD and finger artery dilation by PAT were compared between interventions and groups.

Results:
No differences in NO function in a conduit artery were seen. In contrast, in resistance vessels, high salt diet increased vasodilation in both POTS and healthy subjects (figure). In addition, POTS patients have greater vasodilation than healthy subjects during both low and high salt diets (p=0.036 and 0.033 for high and salt diets respectively).

Conclusions:
POTS patients may have an exaggerated NO response to reactive hyperemia in resistance vessels, but not in conductance vessels. This excessive vasodilation could contribute to POTS symptoms on standing.
Abstract P196: Effect of Combined Oral Intake of Ginger Extract and Rice Vinegar on Blood Pressure with 2-kidney,1-clip Renovascular Hypertensive Rats
Authors: Saito Natsumi , Segawa Yukiko , Maruyama Saki , Yamaoka Ayuna , Hashimoto Hiroko , Osera Tomoko , Kurihara Nobutaka
Published: July 2021

Objective:
Ginger is widely used as traditional Asian herbal medicine. Ginger has the same pungent ingredient as chili and vanillyl. We showed that administration of capsaicin to renovascular hypertension (RH) model rats increased endothelial nitric oxide (NO) synthase (eNOS) mRNA expression and NO production, and suppressed blood pressure (BP). Traditionally in Japan, ginger is pickled and eaten. Ginger and vinegar each are supposed to have an effect of suppressing an increase in BP in RH rats. The aim of this study was to investigate the effect.

Method:
Male Sprague-Dawley rats (6wks) were treated with sham operation (SHAM) as controls or clipping the left renal artery (2K1C) as RH model. After surgery, the rats started receiving a control diet (C) or a diet with 0.08% (w/w) of Ginger Extract (GE) for 6 weeks, and a tap water (W) or a water with 4.5% (v/v) rice vinegar (V). The systolic BP (SBP) was measured by a tail-cuff method every week. At the end of the protocol, the mean arterial BP (MAP) was measured under anesthesia. Then, the aortas were removed for extracting mRNA. mRNA for angiotensin type 2 receptor (AT
2
) and eNOS was evaluated by real-time RT-PCR.

Results:
Through the experiment period, SBP was significantly effects in time, model (SHAM vs 2K1C), diet (C vs GE) , timeхanimal (
P
<0.001, each) and water (W vs V) (
P
<0.05). At the end of the protocol, 2K1C-C+W was higher in SBP than SHAM-C+W (176 ± 6 vs 138 ± 1 mmHg,
P
<0.05). 2K1C-GE+W showed lower SBP (150 ± 2 mmHg) than -C+W (
P
<0.05). SBP was not significantly different in 2K1C-GE+V (149 ± 4mmHg) from in -GE+W. The observations in MAP were similar to those in SBP. AT
2
R mRNA expression showed significant effects in model (
P
<0.05) : the mRNA in 2K1C-C+W (0.9 ± 0.2) was significantly greater than in SHAM-C+W (0.4 ± 0.1) (
P
<0.05). There were no significant differences among the 2K1Cs: -C+W, -C+V (0.9 ± 0.1), -GE+W (0.8 ± 0.1) and -GE+V (0.9 ± 0.2). eNOS mRNA expression showed significant effects only in diet (CTL vs GE,
P
<0.05), but not in water and any interactions.

Conclusion:
Continuous ingestion of GE and V may suppress BP increase in 2K1C, respectively. Simultaneous ingestion of GE and V showed no enhanced effects compared to GE or V solo ingestion in 2K1C. The roles of eNOS and AT
2
R in the mechanism did not become clear in this study.
Abstract P199: Higher Circulating Triglycerides are Independently Associated With Greater Short-term Blood Pressure Variability in Middle-aged/older Adults With Obesity
Authors: Luehrs Rachel E , Kalil Graziela Z , Holwerda Seth W , Wooldridge Nealy A , Fiedorowicz Jess G , DuBose Lyndsey E , Rader Melissa M , Pierce Gary L
Published: July 2021

Elevated short-term (24 hour) blood pressure variability (BPV) is associated with subclinical target organ damage and cardiovascular disease (CVD) among middle-aged/older (MA/O) adults with hypertension and obesity. Circulating total cholesterol (TC), low-density cholesterol (LDL-C) and triglycerides (TGs) increase with human obesity and are independent risk factors for CVD. In addition, BPV is increased in mouse models of hyperlipidemia and is normalized with statins. However, whether higher circulating lipoproteins independently contribute to greater short-term BPV among adults with obesity remains unclear. We hypothesized that higher LDL-C, TGs and lower high-density lipoprotein (HDL-C) would be associated with greater short-term BPV among individuals with obesity. Fasting plasma lipids and 24 hour ambulatory BP monitoring were assessed in fifty-six MA/O adults with obesity defined as body mass index (BMI) ≥ 30 kg/m
2
(56% F; age 54±7 yrs; BMI, 38.2±5.6 kg/m
2
) and at least one other CVD risk factor. There was a significant relation between 24 hour systolic BPV and TC (r=0.30, P=0.03), TGs (r=0.34, P=0.01) and LDL-C (r=0.25, P=0.059), but not HDL-C (r=-0.07, P=0.61). Interestingly, these findings remained significant after adjusting for age, sex, BMI and 24 hour systolic BP (TC: r=0.34, P=0.01; TGs: r=0.39, P<0.01; LDL-C: r=0.31, P=0.03) but HDL-C remained non-significant (r=-0.16, P=0.27). In contrast, other cardiometabolic risk factors such as fasting glucose, insulin, c-reactive protein concentrations, carotid-femoral pulse wave velocity and HOMA-IR were not associated with 24 hour systolic BPV. In a multiple linear regression model that included age, sex, BMI, 24 hour systolic BP, TGs and LDL-C, only fasting TGs (β=0.02 ± 0.01, P=0.02) were a significant correlate of 24 hour systolic BPV (Model R
2
=0.24, P=0.03). Results were the same if TC was substituted for LDL-C in the model. In conclusion, higher plasma TC, LDL-C and TGs are associated with greater 24 hour BPV among MA/O adults with obesity with only TGs being independently associated with BPV. These data suggest that greater variability in BP among MA/O adults with obesity is mediated in part through circulating TGs suggesting that TGs may be a therapeutic target to modify short-term BPV.
Abstract P229: cGMP Stimulates Ubiquitination and Degradation of NKCC2 in Thick Ascending Limbs
Authors: Ares Gustavo R
Published: July 2021

NaCl reabsorption by the thick ascending limb (TAL) is mediated by the Na/K/2Cl cotransporter (NKCC2). Nitric oxide inhibits NaCl reabsorption in TALs by increasing the second messenger cyclic guanylate monophosphate (cGMP) and decreasing NKCC2 in the apical plasma membrane. Recently, we showed that internalized NKCC2 is constitutively degraded (0.33%/min). Protein degradation regulates channels and transporters activity along the nephron. However, whether NKCC2 is regulated by lysosomal degradation or ubiquitin-proteasomal system remains unknown. We hypothesized that internalized NKCC2 is degraded by the ubiquitin-proteasome system in a process stimulated by cGMP. TAL surface proteins were biotinylated and allowed to internalize. The biotin remaining in surface proteins was stripped away and only internalized NKCC2 measured by Western blot. cGMP enhanced the rate of disappearance of internalized NKCC2 by 83 % and this was blocked by a proteasomal (MG132) but not lysosomal (leupeptin) inhibitor (Control: 0.29 ± 0.04; cGMP: 0.53 ± 0.10 ; cGMP + MG132: 0.10 ± 0.10 ; cGMP + Leupeptin: 0.44 ± 0.06 %/min;
p
< 0.05). In general, protein degradation by the proteasomal system requires ubiquitination of the targeted protein. We found that NKCC2 inmuno-precipitated with ubiquitin. Proteasome inhibition induced the accumulation of ubiquitin-NKCC2 and this was enhanced by cGMP (MG132: 59 ± 14%, MG132+cGMP: 111 ± 25%;
p
<0.05). We concluded that internalized NKCC2 is degraded
via
the ubiquitin-proteasome pathway in a process stimulated by cGMP. cGMP-induced degradation of internalized NKCC2 may contribute to decreased NKCC2 trafficking to the apical membrane therefore decreasing NaCl reabsorption in TALs.
Abstract P253: Relation Between Microvascular Function and Large Artery Stiffness in African American Diabetic Patients
Authors: Adedayo Ajibola M , Eluwole Ayobami , Tedla Fasika , Kremer Arye , Mastrogiovanni Nicole , Afzal Mohammed , Rosenberg Carl , Dreizen Paul , LaRosa John , Boutjdir Mohamed , Banerji MaryAnn , Brown Clinton , Salifu Moro , Bakillah Ahmed , Lazar Jason
Published: July 2021

Rationale:
Diabetes is highly prevalent among African Americans and poses a higher risk for vascular complications in this population. Although socioeconomic factors are well known to influence outcomes, true biologic differences in risk factor vulnerability have been suggested. Vascular complications have been traditionally viewed as either macrovascular (myocardial infarction and stroke) or microvascular (retinopathy, nephropathy, and neuropathy). Better glycemic control is known to improve microvascular but not macrovascular complications. In recent years, there has been a growing appreciation that microvascular dysfunction may promote large artery disease and vice versa. Given this notion of vascular “cross-talk” and since subclinical dysfunction is known to precede target organ damage, the objective of this study was to determine whether subclinical microvascular dysfunction is related to large artery stiffness.

Methods:
A total of 141 patients with type II diabetes were recruited from our outpatient clinics over a 6 month period. Medical information was obtained via patient interview and electronic medical record review including laboratory results. Microvascular function was assessed by the 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 and 90% had dyslipidemia. 15% had chronic kidney disease. Mean HbA1C levels were 8.1
+
2.2%. For the entire group, VRI was significantly correlated with PWV (r=.27, p=.002). On multivariate analysis, VRI was independently associated with PWV (β=-1.0, p=.001) and a trend towards an association with HbA1c (β =.07, p=.09) after adjusting for traditional cardiovascular risk factors.

Conclusions:
Among African Americans with diabetes, subclinical microvascular dysfunction is significantly correlated to large artery stiffness and possibly to glycemic control. Further study is needed to clarify mediating factors of these relationships.
Abstract P270: First Trimester Elevation in Circulating Endothelin-1 and Arterial Stiffness are Predictive of Late Pregnancy Preeclampsia
Authors: Betz Alexandria M , Pierce Gary L , Santillan Donna A , Devor Eric J , Scroggins Sabrina M , Brandt Debra S , Stroud Amy K , Thomas Kellie M , Holwerda Seth W , O’Deen Allison L , DuBose Lyndsey E , Grobe Justin L , Sigmund Curt D , Santillan Mark K
Published: July 2021

Preeclampsia (PE) is characterized by late pregnancy hypertension and proteinuria. PE causes significant morbidity for the maternal-fetal unit. Circulating endothelin-1 (ET-1), a potent vasoconstrictor, is elevated at the time of diagnosis of human PE. In addition, women with PE demonstrate arterial stiffness as early as the end of the first trimester. However, it is unknown if arterial stiffness is associated with a first trimester elevation in ET-1 and post-delivery placental ET-1. We hypothesized that 1) first trimester plasma ET-1 is elevated and is associated with arterial stiffness in women who develop PE; 2) first trimester ET-1 is predictive of PE; and 3) placental ET-1 is increased in PE. To address these questions, we performed a nested case-control study in women at risk for PE. First trimester plasma ET-1 was measured via ELISA; aortic stiffness and carotid beta-stiffness (CβS) were measured by carotid-femoral pulse-wave velocity (CFPWV) and carotid tonometry/ultrasound, respectively. While the maternal age of controls (n=126; age 30 ± 0.45 years) and PE (n=15; age 31 ± 1.3 years) were similar, the PE group had a higher first trimester BMI (35 ± 3 vs. 29 ± 1 kg/m
2
, p = 0.01), systolic (125 ± 2 vs. 113 ± 1 mmHg, p< 0.01) and diastolic blood pressure (68 ± 2 vs. 60 ± 1 mmHg, p< 0.01) compared with controls. In addition, first trimester plasma ET-1 (2.7 ± 0.4 vs. 2.0 ± 0.2 pg/mL, p < 0.01), CFPWV (7.2 ± 0.5 vs. 6.1 ± 0.2 m/s, p = 0.016), and CβS (8.4 ± 1.9 vs. 6.3 ± 0.3, p = 0.055) were higher in the PE group. Consistent with previous studies, third trimester plasma ET-1 was elevated in the PE group (2.9 ± 1.1 vs. 1.6 ± 0.1 pg/mL, p < 0.01) which paralleled a 2.5 fold increase in placental decidual ET-1 mRNA (p < 0.0001). ROC analyses showed that first trimester plasma ET-1 (AUC=0.71, p < 0.001) and CFPWV (AUC=0.70, p=0.014) were predictive of PE. This study supports the novel concept that elevated ET-1 in preeclampsia begins early in the first trimester and is associated with premature arterial stiffness. Further, these novel data suggest that ET-1 may play an important role in the first trimester prediction and pathogenesis of preeclampsia.
Abstract P275: Peripheral 18-oxocortisol Concentration Reflects Intra-tumoral Activities of Both CYP11B1 and CYP11B2 in Aldosterone-producing Adenoma
Authors: Tezuka Yuta , Yamazaki Yuto , Kitada Masaaki , Nezu Masahiro , Morimoto Ryo , Kudo Masataka , Igarashi Yasuhiro , Nakamura Yasuhiro , Ito Sadayoshi , Dezawa Mari , Sasano Hironobu , Satoh Fumitoshi
Published: July 2021

Background:
The 18-oxocortisol (18oxoF) is an expected biomarker for detection of patients with aldosterone-producing adenoma (APA) among patients with primary aldosteronism. However, the mechanism of 18oxoF synthesis in APA and differences of the production amount among APAs remain unclear.

Objective:
To examine the associations between peripheral 18oxoF concentration (p18oxoF) and clinical or pathological parameters in APA cases.

Method:
We included 48 APA cases who were diagnosed in our center. The p18oxoF and tissue steroid levels were measured by highly sensitive LC-MS/MS measurement. We performed immunohistochemical (IHC) staining of 11β-hydroxylase (CYP11B1), aldosterone synthase (CYP11B2) and 17α-hydroxylase (c17) for all the cases. The IHC images were analyzed by HALO digital image software. Moreover, we performed immunofluorescence staining to evaluate the number of CYP11B1/B2 double positive cells (B1/B2 cells).

Result:
The characteristics of 48 cases were as follows; Age 51.7 ±11.8 years; Male 54.2% (26/48); Plasma aldosterone (A) 52.2 ±45.8 ng/dl; Plasma renin activity 0.35 ±0.43 ng/ml/h; Serum cortisol (F) 10.0 ±3.5 μg/dl; Cross-sectional area (CSA) of APA 63.9 ±52.7 mm
2
. The p18oxoF showed a lognormal distribution from 0.19 to 183.13 ng/dl (Median; 11.92 ng/dl). The p18oxoF, a derivative of F, was positively correlated with plasma A (r=0.728, P<0.0001) but not serum F (P=N.S.). However, there were significant correlations between p18oxoF and intra-tumoral A or F levels in 15 frozen APA tissues which were available (A: r=0.750, P<0.01, F: r=0.575, P<0.05). On the other hand, p18oxoF was positively correlated with CSA (r=0.734, P<0.0001). Divided them into low and high groups based on p18oxoF, high group had larger CSA (36.1 ±37.9 vs 91.8 ±51.2 mm
2
, P<0.0001) and immunoreactive positive areas of CYP11B1 (7.9 ±12.5 vs 17.9 ±18.4 mm
2
, P=0.003), CYP11B2 (9.0 ±8.8 vs 20.7 ±9.9 mm
2
, P<0.0001) and c17 (6.1 ±10.1 vs 12.6 ±7.6 mm
2
, P=0.0004) than low group. High group also had more B1/B2 cells than low group (106.5 ±93.1 vs 49.3 ±59.3 /mm
2
, P=0.009).

Conclusion:
These results suggest 18oxoF synthesis could be influenced by the size of APA, which reflects intra-tumoral activities of not only CYP11B2 but also CYP11B1, a rate-limiting enzyme of F.
Abstract P314: Effect of Sex on Blood Pressure and Endothelial Dysfunction in the Novel BPH2 Mouse Model of Hypertension
Authors: Didion Sean P
Published: July 2021

Very little is known regarding blood pressure and endothelial function between the sexes in the hypertensive BPH2 mouse. Thus, the first goal was determine whether blood pressure and endothelial function are significantly different between male and female BPH2 mice. Information regarding the role of the renin-angiotensin system in the BPH2 mouse is also limited; therefore the second goal was to determine the role of the renin-angiotensin system by treating BPH2 mice with captopril for 4 weeks. Systolic blood pressure (SBP) was significantly elevated (P<0.05) and yet comparable (P>0.05) in male and female BPH2 mice and averaged 140±3 and 136±3 mmHg, respectively, whereas, in control mice SBP averaged 112±4 mmHg. Endothelial responses to acetylcholine in carotid artery were markedly impaired (P<0.05) and to a similar degree in male and female BPH2 mice as compared to controls. Captopril treatment was associated with a significant (P<0.05) reduction in blood pressure of 35±7 and 43±4 mmHg in male and female BPH2 mice, respectively. Captopril also resulted in an improvement of endothelial responses in male and female BPH2 mice. These findings demonstrate that male and female BPH2 mice are equally hypertensive and both sexes are characterized by endothelial dysfunction. In addition, the renin-angiotensin system may contribute to both hypertension and endothelial dysfunction in this model. Taken together, our data define the BPH2 mouse as an important model to compare and contrast the effects of hypertension between the sexes. Supported by NIH HL-107632.
Abstract P317: Sex Differences in the Hemodynamic and Sympathetic Responses to Centrally Administered Tumor Necrosis Factor-α in Rats
Authors: Wei Shun-Guang , Yu Yang , Felder Robert B
Published: July 2021

Introduction:
Accumulating evidence indicates that sex differences exist in the clinical and experimental outcomes of various cardiovascular diseases. In addition to its protective effect on renin-angiotensin system activity, estrogen has an anti-inflammatory influence. The central actions of pro-inflammatory cytokines (PICs) contribute significantly to cardiovascular and autonomic dysfunction in hypertension and heart failure. In male adult rat, central administration of PICs induces substantial increases in blood pressure (BP), heart rate (HR) and renal sympathetic nerve activity (RSNA), and blocking PICs reduces sympathetic excitation in experimental models of hypertension and heart failure. Whether PICs have similar central sympatho-excitatory effects in the female rat remains unknown.

Hypothesis:
We hypothesized that female rats may be protected from the central cardiovascular and autonomic effects of PICs.

Methods:
Urethane anesthetized male and female Sprague Dawley rats (10-12 weeks) underwent an intracerebrovascular (ICV) injection of the prototypical PIC tumor necrosis factor-α (TNF-α, 100 ng). BP (mmHg), HR (beats/min) and RSNA (% change) responses were continuously recorded for 4-5 hours.

Results:
In male rats (n=6), ICV TNF-α induced a dramatic and long-lasting increase (*p<0.001 vs. baseline) in BP (23.1 ± 2.5*), HR (82 ± 8*) and RSNA (109.5 ± 4.3 %*), that began within 20-30 mins and peaked at 90-120 mins after ICV injection. In the female rats (n=6), ICV TNF-α elicited significantly (p<0.05) smaller increases (*p<0.001 vs. baseline) in BP (14.8 ± 1.8*), HR (55 ± 6*) and RSNA (78.5 ± 6.3*), compared with the male rats.

Conclusion:
These data demonstrate a sex difference in the cardiovascular and sympathetic responses to centrally administered PICs. Whether the observed differences can be explained by an estrogen effect on TNF-α signaling per se or by an estrogen effect on TNF-α-induced renin-angiotensin activity remains to be determined. However, a reduced response of female rats to central inflammation may be an important contributor to sex differences in pathophysiology of hypertension and heart failure.
Abstract P327: Inter-individual Difference in Sensitivity of Aversive Salt Taste
Authors: Isomura Minoru , Nabika Toru
Published: July 2021

Salt is an indispensable nutrient, but excessive amount of salt intake becomes harmful. In order to prevent excessive intake of salt, there is a mechanism to feel high concentration salt as an aversive taste. In recent years, it has clarified that high concentration salt stimulates different taste buds from those sensed by low concentration of salt. In this study, we analyzed relationship between inter-individual difference in sensitivity of aversive salt taste and amounts of daily salt intake. After obtained written informed consent, 1,254 individuals who came annual health checkups were recruited for this study. To test aversive salt taste, five different saline solutions (0.25%, 0.5%, 1%, 1.5%, 2%) were prepared. They started drinking of salt solution from low to high concentration of solutions. In each trial of drinking the salt solution, they were asked whether they were capable of drinking it. When they felt to avoid drinking the salt solution, salt concentration of the solution was recorded as sensitivity of aversive salt taste. Amount of daily salt intake was estimated by using spot urine and by dietary questionnaire. As a result, 784(63%) and 267(21 %) of individuals felt up to the 1.5% and 2% of salt solution as aversive salt taste, respectively. Hence, their sensitivities of aversive salt taste were 1.5% or 2% of salt solutions, respectively. However, resting 203(16%) of individuals showed capability to drink 2% of salt solution. Although we compared sensitivities of aversive salt taste of each individual and daily salt intake that estimated by urine, there were no difference between them. However, daily salt intake that estimated by dietary questionnaire showed clear relationship, in which individuals with the high sensitivities of aversive salt taste consumed the more amount of salt.
Abstract P328: Angiotensin II Inhibits the High Salt-induced Production of Sphigosine-1-phosphate in the Renal Medulla
Authors: Wang Weili , Hu Junping , Li Pin-Lan , Poklis Justin L , Li Ningjun
Published: July 2021

We have previously shown that sphigosine-1-phosphate (S1P) produces natriuretic effects via activation of S1P receptor 1 in the renal medulla and that this natriuretic effect may be through inhibition of epithelial sodium channel. The present study examined the expression of the enzymes that produce S1P in the renal medullary tissue and tested the hypothesis that angiotensin II (ANG II) reduces the expression of S1P-producing enzyme and thereby the levels of S1P in the renal medulla. Male adult C56BL/6 mice, 10-12 weeks old, were treated with a low salt diet (LS, 0.4%), high salt diet (HS, 4% NaCl) or HS + ANG II (600ng/kg/min, S.C.) for 10 days. A high salt diet increased the level of S1P, whereas ANG II significantly inhibited the HS-induced increase of S1P levels in the renal medullary tissue. The levels of S1P were 6.6 ± 0.34, 11.4 ± 1.33 and 3.5 ± 0.49 pmol/mg of tissue in LS, HS and HS + ANG II group, respectively. There were no difference in the levels of sphingosine kinase 1 (SPHK1), the enzyme that produces S1P by phosphorylating sphingosine, among the different groups of mice by Western blot analysis. However, a high salt diet increased the protein levels of acid ceramidase (ACDase), an upstream enzyme that produces sphingosine, the substrate for SPHK1. This HS-induced increase in ACDase was inhibited by ANG II. The relative protein levels of ACDase were 1.0 ± 0.07, 1.4 ± 0.07 and 0.17 ± 0.11 in LS, HS and HS + ANG II group, respectively. These results demonstrated that a high salt diet increased the levels of S1P in the renal medulla, probably by increasing the level of one of the S1P-producing enzymes ACDase, and that ANG II reduced the levels of ACDase and S1P in the renal medulla. Given the diuretic effect of S1P, ANG II-induced reduction of S1P production in the renal medulla may be a mechanism contributing to the sodium retention and hypertension associated with excessive ANG II. (Support: NIH grant HL89563)
Abstract P331: Angiotensin-salt Hypertension Requires Ouabain-sensitive Na
+
Pumps
Authors: Chen Ling , Lingrel Jerry B , Hamlyn John M , Blaustein Mordecai P
Published: July 2021

Dietary salt is a major factor in the pathogenesis of essential hypertension (EH), but the underlying links are unresolved. Animal models indicate that angiotensin (Ang) II and high dietary salt (HS) are convergent signals that act via the brain to elevate blood pressure (BP). Low-dose sc Ang II+HS is a common model for EH. We tested the Na
+
pump ouabain binding site’s role in this model because it is crucial in some other hypertension models (e.g., ACTH and Nedd4-2-knockout +HS). Mice that express Na
+
pumps with a mutant, ouabain-resistant α2 catalytic subunit (α2
R/R
; cation transport is normal), and wild type (WT), ouabain sensitive controls (α2
S/S
) were studied. [80-90% of rodent artery myocyte Na
+
pumps are ouabain-resistant (α1
R/R
); only 10-20% are α2.] BP was measured by telemetry. First, 3 basal 24 hr BPs were recorded. Osmotic 4-week minipumps were then implanted sc in all mice to deliver vehicle (saline; Expt. #1,3), or 400 (Expt. #1,2) or 800 (Expt. #3) ng/kg/min Ang II; simultaneously, in Expt. #2, the diet was switched from 0.4% (standard) to 2% NaCl (HS). BPs were monitored every 3-4 days for up to 4 weeks. Also, in Expt. #2, on day 21, all mice received 2 ip injections, 4 hrs apart, of 10 mg/kg DigiFab, Fab fragments that immuno-neutralize ouabain, while BP was continuously monitored; on day 23, the mice received 2 ip injections of CroFab, anti-crotalus toxin (‘control’) Fab fragments.

Results:
1. Basal mean BP (MBP) was 10±2 mm Hg higher in α2
R/R
than in WT mice (
P
<0.01;
n
=21 & 29; ANOVA). 2. In WT mice, 400 ng/kg/min sc Ang II and Ang II+HS raised MBP by 15±1 and 34±1 mm Hg, respectively (
P
<0.01;
n
=7-8; ANOVA). 3. The MPB elevation in Ang II+HS α2
R/R
(17±2 mm Hg) was only half that in WT mice (
P
<0.01;
n
=7 each; ANOVA). 4. DigiFab rapidly (<1 hr) reduced MBP by 14±2 mm Hg in Ang II+HS hypertensive WT mice (
P
<0.001;
n
=7; T-test), but not in α2
R/R
mice (
P
<0.01;
n
=7 each; ANOVA); CroFab did not lower MBP in either strain. 5. 800 ng/kg/min sc Ang II elevated systolic BP by 55±3 mm Hg in WT mice, but by only 37±3 mm Hg in α2
R/R
mice (
P
<0.05;
n
=3-5; ANOVA).

Conclusions:
Ouabain-sensitive α2 Na
+
pumps and their endogenous ligand are both required for full expression of low-dose Ang II-salt hypertension. Ouabain-sensitive α2 pumps apparently also contribute to high-dose Ang II-hypertension.
Abstract P338: Long Term End Organ Inflammation and Dysfunction in Mice After Angiotensin II Induced Hypertension
Authors: Chen Wei , Xiao Liang , Harrison David G
Published: July 2021

Hypertension is associated with vascular and renal inflammation leading to organ dysfunction and injury. Although reduction of hypertensive stimuli or antihypertensive therapy can lower blood pressure, it is unclear if inflammation persists beyond the initial hypertensive stimulus. We sought to examine the hypothesis that a short-term hypertensive insult leads to ongoing inflammation and end organ dysfunction. C57BL/6 mice received a subcutaneous infusion of angiotensin II (490 ng/kg/min) or a sham infusion via osmotic minipumps for two weeks. The minipumps were then removed, and the mice were allowed to recover for two months. To evaluate renal function, mice received a challenge of normal saline equal to 10% of each body weight via intraperitoneal injection and the urine excreted in the subsequent 4 hours was measured. Whereas sham-treated mice excreted 88±7%, mice that had received ang II 2 months earlier excreted only 51±9% of the injected volume (p<0.05). Moreover, albuminuria was doubled in the mice that had received prior ang II infusion (0.55 ± 0.1 vs 0.26 ± 0.1 μg/ml, p < 0.05). After sacrifice, the renal and aortic samples of both groups of mice were analyzed by flow cytometry. We found that the numbers of total leukocytes (CD45
+
), total T lymphocytes (CD3
+
) and monocytes/macrophages (F4/80
+
) were 4 to 5 times higher in aortas and 45 to 70% higher in the kidneys even after two months following ang II infusion compared to sham-treated mice. We have previously shown that isolevuglandin-protein adducts in antigen presenting cells are immunogenic, and we found that these were persistently in MerTK
+
/CD64
+
macrophages in the aorta mice that had received ang II 2 months earlier. There was also striking perivascular fibrosis in mice that had received prior ang II, but not in sham infused mice. These data indicate that a persistent inflammatory process, accompanied with renal dysfunction and aortic fibrosis continues for a prolonged period of hypertension. Efforts to ameliorate this might reduce the long-term risk renal and vascular disease.
Abstract P342: Sirt 1 on Endothelial Dysfunction in Small Arteries From Obese Patients
Authors: Virdis Agostino , Masi Stefano , Nannipieri Monica , Guarino Daniela , Anselmino Marco , Colucci Rocchina , Duranti Emiliano , Blandizzi Corrado , Taddei Stefano
Published: July 2021

Obesity is associated with endothelial dysfunction, characterised by a reduced nitric oxide (NO) bioavailability due to increased reactive oxygen species (ROS). Sirtuins, and more specifically Sirt-1, are enzymatic proteins involved in regulation of glucose metabolism, inflammation and intracellular levels of ROS. This study aimed to determine the role of Sirt-1 in regulating NO bioavailability of small resistance arteries isolated from subcutaneous tissue of obese patients. 10 subjects (5 with severe obesity, Ob; 5 normal weight controls, Ctrl) underwent biopsy of subcutaneous adipose tissue during laparoscopic bariatric surgery. Function of small arteries was assessed with pressure micromyography. Endothelial-dependent vasodilation (VDep) and NO production was assessed by acetylcholine (ACh, 0,001-100μM), with and without pre-incubation with L-NAME (100μM). The influence of sirtuins on NO bioavailability was assessed repeating Ach with a selective Sirt-1 agonist (SRT-1720, 1μM), alone or plus L-NAME. Ob showed a reduced response to Ach vs Ctrl (P<0.001), associated with a reduced inhibition of L-NAME on Ach (Ob: P=0.002; Ctrl: P<0.001). SRT-1720 improved the VDed induced by Ach (P<0.001) in Ob, although it did not reach values from Ctrl. The simultaneous incubation with L-NAME and SRT-1720 before Ach stimulation abolished the VDed obtained with the incubation of SRT-1720 in the Ob group (SRT-1720 vs SRT-1720+L-NAME: P<0.001). In small arteries of Ob, stimulation of Sirt-1 activity partially restores endothelial function due to an improved NO bioavailability.
Abstract P348: Immunosuppression with Cyclophosphamide Attenuates Renal Injury but Not Hypertension in an Experimental Model of Autoimmune Disease
Authors: Wolf Victoria L , Taylor Erin B , Ryan Michael J
Published: July 2021

Cardiovascular disease is the major cause of mortality among patients with the autoimmune disorder systemic lupus erythematosus (SLE). Our laboratory previously showed that immunosuppression with mycophenolate mofetil, a common therapy in patients with SLE, attenuates the development of hypertension in an experimental model of SLE. Cyclophosphamide is another common therapy for patients with SLE that has contributed to improved disease management; however, its impact on the development of hypertension associated with SLE is not clear. We tested whether treatment with cyclophosphamide (25 mg/kg, once/week, IP injection) for four weeks attenuates hypertension in an established female mouse model of SLE with hypertension (30 week old NZBWF1 females). Plasma anti-dsDNA IgG levels, pathogenic for the disease, were lower in cyclophosphamide-treated SLE mice compared to vehicle-treated SLE mice (161 ± 43, n=19 vs 501 ± 114, n=21, p<0.05 units/mL), suggesting efficacy of the therapy to suppress aberrant immune system function. Mean arterial pressure (MAP) was assessed by carotid artery catheters in conscious mice. Treatment did not attenuate the development of hypertension (136 ± 3 mmHg, n=15) when compared to vehicle treated SLE mice (131 ± 4 mmHg, n=13, p=0.69]; however, urinary albumin excretion (72.2 ± 68 mg/day, n=21, mg/day) was lower in cyclophosphamide treated animals (0.06 ± 0.02 mg/day, n=13). Corresponding with the reduction in autoantibodies, preliminary data suggest that cyclophosphamide treatment lowered circulating CD45R
+
B cells (15.87 ± 8.49%, n=10 vs 26.96 ± 4.72%, n=8). Paradoxically, circulating CD11b
+
Ly6G
+
neutrophils were increased in cyclophosphamide treated SLE mice compared to vehicle treated (39.26 ± 4.92%, n=10 vs 20.58 ± 6.01%, n=8, p=0.06). These data suggest that cyclophosphamide treatment attenuates autoantibody production and renal disease during SLE, but that the potential to impact MAP may be blunted by the increase in circulating neutrophils.
Abstract P352: Prostaglandin E2 Reduces Mitochondrial Function in Adult Mouse Cardiomyocytes
Authors: Harding Pamela , Bryson Timothy D , Datta Indrani , Wang Yun , Levin Albert
Published: July 2021

Hypertension is a leading cause of heart failure and both conditions are characterized by increased prostaglandin E2 (PGE2) which signals through 4 receptor subtypes (EP1-EP4) to elicit diverse physiologic effects. We previously reported that cardiomyocyte-specific deletion of the EP4 receptor results in a phenotype of dilated cardiomyopathy in male mice that is characterized by reduced ejection fraction. Subsequent gene array on left ventricles from these mice, coupled with Ingenuity Pathway Analysis (IPA) demonstrated that genes differentiating WT mice and EP4 KO mice with low ejection fraction were significantly overrepresented in mitochondrial (p=2.51x10
-28
) and oxidative phosphorylation (p=3.16 x10
-30
) pathways. We therefore hypothesized that PGE2 could reduce mitochondrial function. To test this hypothesis, we used isolated mouse cardiomyocytes (AVM) from 16-18 week old male C57Bl/6 mice and treated them with 1 μM PGE2 for various times. Mitochondrial gene expression was examined using a RT-profiler kit for mitochondrial energy metabolism, complex I activity with a spectrophotometric assay, ATP levels with a bioluminescence assay and mitochondrial membrane potential using JC-1 staining. Treatment of AVM with PGE2 for 4 hrs reduced expression of multiple genes from mitochondrial pathways including sub units of mitochondrial NADH dehydrogenase ubiquinone flavoprotein (Nduf), a component of complex I. In accord with the mRNA data, Complex I activity was reduced by 50% (p < 0.05) by 4 hr treatment with PGE2, from 1.32 ± 0.36 to 0.66 ± 0.08 mOD/min. Cytochrome c oxidase subunit 8 (Cox8c) mRNA was also reduced from a control value of 1.00 to -1.75 ± 0.20 (p < 0.005) after PGE2 treatment. Immuno-fluorescence showed that JC-1 aggregates were reduced after 1 or 3 hr treatment with either 1 μM PGE2 or the EP3 agonist, sulprostone, suggesting reduced mitochondrial membrane potential. Subsequent experiments also showed that ATP levels were reduced 16% from 11.18 ± 0.71 nmol to 9.39 ± 0.83 nmol after treatment with sulprostone for only 1 hr. Taken together, these results suggest that increased PGE2 in hypertension may contribute to impaired mitochondrial function and provide yet another link between inflammation and cardiac dysfunction.
Abstract P355: Prevalence of Diastolic Dysfunction With Preserved Left Ventricular Systolic Function in Hypertensive and Type II Diabetic Patients in a Large Community Practice in Greenville, South Carolina
Authors: Saxena Naveen R , Saxena Juhi , Saxena Anju , Srivastava Vinita
Published: July 2021

Diastolic dysfunction with preserved left ventricle systolic function is a major cause for adverse cardiovascular events in hypertensive and Type II Diabetic patients. Review of medical literature reveals prevalence of diastolic dysfunction in the range of 20 to 60 percent in hypertensive and diabetic patients. The aim of this study is to examine prevalence of diastolic dysfunction with normal systolic function in hypertensive and Type II Diabetic patients in a community practice. This is a retrospective chart review of 3085 hypertensive and 899 type II Diabetic patients. All patients underwent 2D color Doppler studies for the evaluation of diastolic dysfunction using American Society of Echocardiography criteria. E wave velocity, A wave velocity, E to A ratios, and deceleration time were measured. Patients with known systolic heart failure were excluded from the study. The age distribution of the patients in the study ranged from 45 to 85 years with a mean age of 65 years. The sex distribution of the hypertensive patients were 45% male and 55% female. The sex distribution of Type II Diabetic patients were 44% male and 56% female. Given the data, we conclude prevalence of diastolic dysfunction in hypertensive patients is 29% and prevalence of diastolic dysfunction in type II Diabetic patients is 33%. Echocardiography is an excellent tool to risk stratify hypertensive and type II Diabetic patients. Aggressive management of this high-risk group may reduce cardiovascular mortality and morbidity.
Abstract P380: Blood Pressure Control Rates When Measured in a Specialty Clinic versus Primary Care Within a Large Integrated Health System
Authors: Billups Sarah J , Saseen Joseph J , Vande Griend Joseph P , Schilling Lisa M
Published: July 2021

Background:
Treating patients with hypertension to goal blood pressure (BP) values is a pillar of cardiovascular risk reduction. Its importance is prominent in quality metrics used in public reporting, accreditation, and value-based reimbursement programs. We hypothesized that BP measurements performed in a specialty (SP) clinic would be less likely to be at goal compared to when measured in a primary care (PC) clinic.

Objective(s):
Identify differences in rates of BP control for patients with hypertension using measurements performed in a SP clinic versus a PC clinic.

Methods:
Clinic-based BP measurement performed in 2016 for adults in Colorado’s UCHealth Clinically Integrated Network with a hypertension diagnosis were included. The most recent measurement for each patient was classified as controlled (<140/90 mm Hg) or uncontrolled, and from a PC or SP clinic. Logistic regression was used to calculate the OR and 95% CI for the likelihood of having controlled BP in a SP versus PC clinic, controlling for age and gender. A secondary analysis of patients with measurements done in both clinic types used conditional logistic regression to compare the likelihood of control based upon each patient’s most recent SP versus PC measurement.

Results:
Of the 86,632 hypertensive adults with BP measurements in 2016, 51% (43,850/86,632) had their most recent measurement performed in a SP clinic. Control rates for BP were lower in SP versus PC clinics (63% (27,771/43,850) vs 67% (28,843/42,782), OR=0.84 (95% CI 0.81-0.86)). Among patients with measurements performed in both clinics in 2016, control rates for the same patients were lower in SP compared to PC clinics (63% (17,900/28,455) vs. 71% (20,077/28,455), OR=0.64 (0.61-0.66)). This pattern persisted even in patients whose measurements were done within 7 days of each other (63% (1,634/2,593) vs 69% (1,781/2,593), OR=0.70 (0.61-0.80)).

Conclusions:
BP measurements performed in SP clinics are significantly less likely to be controlled compared to those in PC clinics, even between measurements performed on the same patient within a short time. This has significant treatment and quality measurement implications for health systems. Investigation is needed to identify underlying reasons and to resolve discrepancies.
Abstract P391: Local Ionotropic Glutamate Receptors Contribute to the Sympathoexcitatory Effects of Leptin in the Paraventricular Nucleus
Authors: Shi Zhigang , Brooks Virginia L
Published: July 2021

In male rats, intracerebroventricular infusion of leptin increases sympathetic nerve activity (SNA), and this response can be partially reversed by nanoinjections of the ionotropic glutamate receptor (iGluR) blocker, kynurenate (KYN), and also the melanocortin type 3/4 receptor (MC3/4R) antagonist, SHU9119, into the hypothalamic paraventricular nucleus (PVN). These results suggest that leptin increases SNA in part via activation of PVN iGluR and MC3/4R. More recently, we found that injection of leptin directly into the PVN also dose-dependently increases SNA. However, the role of PVN iGluR or MC3/4R in this response is unknown. Therefore, we determined if PVN KYN (2.7 nmol in 60 nL) or SHU9119 (30 pmol) reverses the increases in lumbar SNA (LSNA), mean arterial pressure (MAP), and heart rate (HR) evoked by bilateral PVN nanoinjections of leptin (30 ng in 60 nL; n=6) or, as a control, of artificial cerebrospinal fluid (aCSF; n=4). After ninety min, PVN leptin increased (all P<0.05) LSNA to 164±10 % of control, MAP from 106±5 to 118±5 mmHg, and HR from 360±12 to 399±16 bpm. Then, PVN injections of KYN decreased (all P<0.05) LSNA to 133±8 % control, MAP to 108±4 mmHg, and HR to 379±16 bpm. On the other hand, neither PVN aCSF injections (LSNA, to 105±3 % control; MAP, from 107±8 to 109±7 mmHg; and HR, from 363±31 to 367±20 bpm) nor subsequent PVN KYN injections (LSNA, to 102±5 % control; MAP, to 108±6 mmHg; and HR, to 373±17 bpm) significantly altered these variables. In a second set of animals, while PVN leptin (n=4) elicited similar increases (all P<0.05) in LSNA (to 158±8 % control), MAP (from 101±6 to 113±4 mmHg), and HR (from 367±19 to 405±11 bpm), subsequent SHU9119 had no effects (LSNA, to 151±7 % control; MAP, to 108±5 mmHg; and HR, to 400±14 bpm). Again, no changes in these variables were observed in response to PVN injections of aCSF (n=4), even when followed by SHU9119 (LSNA, to 114±11 and then 110±13 % control; MAP, from 107±7 to 109±8 and then 106±9 mmHg; and HR, 357±31 to 355±24 and then 354± 25 bpm). In conclusion, activation of local iGluR, but not MC3/4R, contribute to the increases in LSNA, MAP, and HR in response to PVN leptin.
Abstract P413: Vascular AT1 Receptors Control Blood Pressure by Augmenting Peripheral Vascular Resistance in Female Mice
Authors: Wolf Erin , Diaz Edward , Azad Hooman , Kupin Aaron , Sparks Matthew A
Published: July 2021

Angiotensin (Ang) II is a major mediator of hypertension pathogenesis and end organ damage. Our understanding of how Ang II elicits these effects on a cellular level continues to evolve. However, the majority of these studies utilize male subjects, despite the awareness that males and females differ in mechanisms of BP regulation. Here we investigated the role of vascular smooth muscle cell (VSMC) AT1A receptors to BP control in female mice. Using Loxp technology and Cre transgenes, we created female mice with cell-specific deletion of AT
1A
receptors in smooth muscle cells (SMKO mice). We found that elimination of these receptors led to a significant (8 mmHg) reduction in baseline BP in female SMKO mice (similar to male SMKO mice) compared with controls (108±2 versus 116±1 mmHg;
P
=0.004) with no effect on modulating sodium sensitivity in female SMKO mice (unlike male SMKO mice in which sodium sensitivity was enhanced). Over a 4-week Ang II infusion, the severity of Ang II
-
dependent hypertension was minimally affected during the first 2 weeks of Ang II infusion, corresponding to no difference in sodium excretion. However, female SMKO mice had a 35% less BP elevation compared to controls over the final 2 weeks of Ang II infusion (SMKO, 20±2 versus control, 30±3 Change in SBP mmHg;
P
=0.01). The acute vasoconstrictor responses to Ang II in the systemic vasculature were reduced (by approximately 50-75%) in female SMKO mice (0.1 μg/kg- SMKO, 3.6±1.2 mmHg versus control, 7.3±0.7 mmHg;
P
=0.01, 0.3 μg/kg- SMKO, 3.3±1.8 mmHg versus control, 12.1 ± 1.3 mmHg;
P
=0.0008, 1.0 μg/kg- SMKO, 8.5±1.9 mmHg versus control, 19.4±2.0 mmHg;
P
=0.002). In addition, the acute Ang II reduction in renal tissue perfusion was nearly eliminated in female SMKO mice. In contrast, we previously reported that in male SMKO mice have a much larger (50%) reduction in BP during chronic Ang II infusion associated with increased natriuresis, but relatively preserved vasoconstrictor response (25% reduction) during acute Ang II infusions. These findings suggest that in female mice, direct actions of AT
1A
receptors in VSMCs are essential for regulation of basal BP and Ang II-dependent hypertension. However, this effect is mediated by reductions in peripheral vascular resistance rather than sodium retention.