bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P193: Angiotensin-(1-7) Acutely Induces Browning of White Adipose Tissue in Mice
Authors: Miller Amanda J , White Melissa , Bingaman Sarah S , Arnold Amy C
Published: December 2018

Angiotensin (Ang)-(1-7) is a beneficial hormone of the renin-angiotensin system that is emerging as a promising target for obesity. We previously showed that chronic Ang-(1-7) treatment attenuates high fat diet (HFD)-induced weight gain in mice by increasing markers of thermogenesis in subcutaneous white adipose tissue (“browning”) to increase energy expenditure. In this study, we tested the hypothesis that Ang-(1-7) could acutely increase adipose thermogenesis in mice. To test this, adult male C57BL/6J mice were placed on a chow diet or 60% HFD for 12 weeks and then received a single subcutaneous injection of Ang-(1-7) (2mg/kg) or saline. This study included 4 groups of mice: chow+saline (n=9), chow+Ang-(1-7) (n=15), HFD+saline (n=8), HFD+Ang-(1-7) (n=10). Core temperature was measured at baseline and at 6 hours after injection. Subcutaneous white and brown adipose tissues were collected 6 hours post-injection. Gene expression of the thermogenic marker uncoupling protein 1 (UCP1) was measured in tissues by quantitative real-time PCR and quantified with 2-ΔΔCT methods. Acutely, Ang-(1-7) did not alter body mass in chow or HFD mice (
P=
0.992). There were 5 chow fed mice (33%) and 2 HFD mice (20%) that did not respond to Ang-(1-7) injection. In the chow diet responders, Ang-(1-7) increased core temperature (36.7±0.2 vs 35.9±0.2°C saline;
P
=0.012) and UCP1 gene expression in subcutaneous white adipose tissue (14.3±5.8 vs 1.9±0.6 saline;
P
=0.050), with no effect in brown adipose tissue (1.7±0.6 vs 1.0±0.2 saline,
P
=0.284). In the HFD responders, core temperature was not altered by Ang-(1-7) (37.8±0.1 vs 37.8±0.1 saline,
P
=0.689). There was a trend towards decreased UCP1 expression in white adipose of HFD-saline mice compared to chow-saline mice (
P
=0.196). Ang-(1-7) significantly increased UCP1 expression in subcutaneous white adipose of HFD responders (6.2±3.1 vs 0.9±0.2 saline,
P
=0.036), with no effect in brown adipose tissue (1.2±0.1 vs saline: 1.7±0.4 saline,
P
=0.402). These data provide further evidence that targeting Ang-(1-7) may be a promising strategy to increase white adipose thermogenesis, an effect that could serve to enhance energy expenditure and restore energy balance in obesity.
Abstract P197: Vacuolar-type H
+
-ATPase Enhances Renal (Pro)renin Expression and Inflammation in Response to High Glucose
Authors: Akhtar Safia , Devitt Rebecca M , Culver Silas A , Li Caixia , Siragy Helmy M
Published: December 2018

(Pro)renin receptor (PRR) is an accessory subunit for vacuolar-type H-ATPase (V-ATPase). Recently, we demonstrated that PRR expression is upregulated in diabetes and promotes renal inflammation. We hypothesized that V-ATPase enhances renal PRR expression and inflammation in response to high glucose. Rat renal mesangial cells (RMCs) were cultured for 3 days in medium containing 25 mM D-glucose (high glucose) for experiments groups and 5 mM D-glucose plus 20 mM L-glucose (normal glucose) for control group. Cells were serum starved for 6 hrs at the end of the 3 day glucose exposure, thereafter 100nM bafilomycin, a V-ATPase inhibitor, treatment was given for 1 hour. At the end of experiments, cells were harvested for total RNA and protein for the determination of PRR and interleukin-6 (IL-6). High glucose significantly (P<0.05) increased mRNA and protein expression of PRR (Fig 1) by 87 (1 vs 1.87) and 52% (1 vs 1.52) respectively, and IL-6 level (Fig 2) by 44% (1 vs 1.44, P<0.05), when compared to their respective normal glucose cells. Bafilomycin treatment attenuated the increase in mRNA and protein expression of PRR by 74 (1.87 vs 0.47, P<0.05) and 45% (1.52 vs 0.83, P<0.05). Similarly, IL-6 mRNA level was also lower by 62% (1.44 vs 0.54, P<0.05) in bafilomycin treated high glucose cells compared to its non-treated counterpart. The present study conclude that V-ATPase contributes to increased renal PRR expression and inflammation in presence of hyperglycemia.
Abstract P202: Targeting the Cardiac Sympathetic Afferent Reflex in Hypertrophic Cardiomyopathy
Authors: Larson Robert A , Lu Yongjun , Benson Christopher J , Chapleau Mark W
Published: December 2018

Sarcomere gene mutations cause hypertrophic cardiomyopathy (HCM). Hallmark features include cardiac hypertrophy, fibrosis, excessive cardiac sympathetic tone, and increased risk of arrhythmias and sudden death. The mechanism(s) of increased sympathetic tone is poorly understood. We hypothesized that cardiac spinal (sympathetic) afferents are sensitized by metabolites (e.g., H+) produced during myocyte energy depletion in HCM, resulting in an enhanced cardiac sympathetic afferent reflex (CSAR). We studied mice with cardiac-specific expression of mutated alpha-tropomyosin (Glu180Gly), an established model of HCM. The CSAR was assessed by measuring mean arterial pressure (MAP) and heart rate (HR) responses to epicardial application of the TRPV1 agonist capsaicin (2.5mM, 2μL) to the left ventricle in anesthetized, ventilated HCM (n=5) and wild-type (WT) littermate control (n=5) mice. Pressor and tachycardic responses to capsaicin were markedly enhanced in HCM mice (Table). Blocking the tonic influence of cardiac afferent activity by epicardial application of the local anesthetic lidocaine (2%) caused small, significant decreases in MAP in both groups of mice; but decreased HR markedly in HCM mice only (n=5) (Table). mRNA expression (qPCR) of acid-sensing ion channel 3 was increased by 2-fold in dorsal root ganglia (T1-T9) of HCM vs. WT mice (n=6 each, P<0.05), suggesting a mechanism for sensitization of cardiac sympathetic afferents in HCM. We conclude that the CSAR is enhanced in HCM mice and speculate that increased activity of cardiac ‘sympathetic’ afferent nerves contributes to increased sympathetic tone, arrhythmias and disease progression in HCM.
Abstract P203: Cumulative Incidence of Orthostatic Intolerance After Bariatric Surgery
Authors: Zhang James B , Tamboli Robyn A , Albaugh Vance L , Kilkelly Donna M , Grijalva Carlos G , Williams David B , Shibao Cyndya A
Published: December 2018

Several retrospective studies describe an orthostatic intolerance (OI) syndrome after bariatric surgery. However, its incidence remains unknown. Using a de-identified registry of 4547 bariatric surgery patients, we identified cases of OI syndrome via structured chart reviews for all subjects who reported new-onset post-operative symptoms of OI. Cases of OI were confirmed using an operational case definition developed by the Vanderbilt Autonomic Dysfunction Center, and available autonomic function test results were examined. The cumulative incidence of post-bariatric surgery OI syndrome was estimated using a life table. Of 741 patients reporting new OI symptoms after surgery, we confirmed the presence of post-bariatric surgery OI syndrome in 85 patients (1.9% of 4547), 14 with severe OI requiring pressor agents. At 5 years post-surgery, follow-up was reduced to 15% (682 of 4547), and the estimated cumulative incidence of OI syndrome adjusted for losses to follow-up was 4.2%. Most OI cases developed during relatively weight-stable months, and weight change was not significantly different between patients with and without OI syndrome (Figure 1). Of OI cases with available autonomic function test data, 52% (11 of 21) showed evidence of impaired sympathetic vasoconstrictor activity. In conclusion, new onset orthostatic intolerance is relatively frequent in the bariatric surgery population. Some patients with OI exhibit evidence of impaired autonomic function. Figure 1. Excess Weight Loss for All Bariatric Operations (solid) vs. Patients with OI Syndrome (dotted). Shaded Areas Represent 95% Confidence Interval. Rug Plot on X-axis Indicates Time of OI Onset per Patient.
Abstract P204: Induction of Endoplasmic Reticulum Stress Upregulates Renin Expression in Immortalized Mouse Hypothalamic N43/5 Cells
Authors: Gomez Javier A , Nakagawa Pablo , Sigmund Curt D
Published: December 2018

Dysregulation of the brain renin-angiotensin system (RAS) has been implicated in many forms of hypertension, including resistant hypertension. However, the mechanism by which the brain RAS is activated is unknown. We have recently published on the activation of brain RAS following disinhibition of renin-a expression after deletion of the brain-specific renin-b isoform. Endoplasmic reticulum (ER) stress, which is an accumulation of unfolded proteins in the ER, has been linked to hypertension as central angiotensin II administration induced ER stress markers. Here, we hypothesize that ER stress induces brain RAS activation by upregulating renin expression in the brain. We first tested if ER stress could induce renin expression
in vitro
in a mouse neuroblastoma hypothalamic cell line, N43/5 cells. After treating cells with ER stressor thapsigargin, a SERCA pump inhibitor, (100nM) for 8 hours we observed transcriptional upregulation of ER stress marker Bip (28.9±5.0 fold increase vs vehicle, p<0.05, n=6). This same treatment also induced an increase in renin expression (13.8±5.8 fold increase vs vehicle, p<0.05, n=6). We have previously shown that deletion of renin-b in the brain causes an increase in renin-a expression specifically in the rostral ventrolateral medulla (RVLM). This data suggests that ER stress may induce RAS activation through a similar renin-a disinhibitory mechanism. Under basal conditions, the RVLM of renin-b knockout mice showed no difference in gene expression of ER stress marker Bip (0.7±0.5 fold increase vs control, p=0.67, n=3-4) or Chop (0.7±0.35 fold increase vs control, p=0.64, n=3-4). This suggests that ER stress may be upstream of renin induction. To test this, we treated C57BL/6 mice with tunicamycin (40mg/mL, 2uL) via intracerebellar ventricle cannula. We detected a positive endogenous renin transcript signal via
in
situ hybridization bilaterally in the medulla in the vicinity of the RVLM (possibly the nucleus ambiguous) from vehicle and tunicamycin treated mice. We are currently testing if ER stress quantitatively increases renin expression using qPCR. These data extend previous studies showing that RAS activation in the brain causes ER stress by suggesting that ER stress may also induce the brain RAS.
Abstract P213: Angiotensin Converting Enzyme Inhibitors (ACEi) Increase Antifibrotic Biomarkers in African American Patients With Hypertension and Left Ventricular Hypertrophy
Authors: Romero Cesar A , Reed Brian , Mathew Shobi , Brodi Aaron , Dawood Rachel , Twiner MIchael , McNaughton Candance , Flack John M , Carretero Oscar A , Levy Phillip D
Published: December 2018

ACEi are first line treatment in hypertension; however, there is controversy regarding the benefit over other antihypertensive drugs. ACEi have pressure independent effects that may make them preferable for certain patients. We aimed to evaluate the impact of ACEi on antifibrotic biomarkers in hypertensive patients with left ventricular hypertrophy (LVH). We conducted a post hoc analysis of a randomized controlled trial where hypertensive African American patients with LVH and vitamin D (VTD) deficiency were randomized to receive standard antihypertensive therapy plus VTD supplementation or placebo. We selected patients who had detectable lisinopril in plasma at one year follow-up and compared them to subjects who did not. The profibrotic marker propeptide of procollagen type I (CPIP) and the antifibrotic markers: matrix metalloproteinase-1 (MMP-1), tissue inhibitor of MMP1 (TIMP-1), the MMP-1/TIMP-1 ratio, telopeptide of collagen type I (CITP) and Ac-SDKP peptide were measured. Sixty six patients were included. Table 1 shows patients characteristics. Patients with lisinopril had lower blood pressure at one-year but no differences were observed in left ventricular mass index (LVMI). The antifibrotic markers Ac-SDKP (3.9±2.6 vs 6.3±2.8; p<0.001), MMP1 and MMP1/TIMP-1 ratio were higher in patients with detectable ACEi (all p<0.05). In a model adjusted for systolic blood pressure, low LVMI (p=0.01), MMP1/TIMP-1 (p=0.02) and Ac-SDKP (p<0.001) levels were associated with lisinopril. We conclude that ACEi increase antifibrotic biomarkers in African Americans with hypertension and LVH, suggesting that they may be offer added benefit over other agents in such patients.
Abstract P219: Site-1 Protease-Derived Soluble (Pro)Renin Receptor Contributes to Salt-Sensitive Hypertension via Activation of the Intrarenal RAS
Authors: Wang Fei , Peng Kexin , Xie Shiying , Yang Kevin , Luo Renfei , Yang Tianxin
Published: December 2018

Dahl salt-sensitive (SS) hypertensive rats exhibit increased expression of renal (pro)renin receptor (PRR) and aberrant activation of the intrarenal renin-angiotensin system (RAS) with unclear functional implication. Recent evidence demonstrates that soluble PRR (sPRR) serves as a potential regulator of tubular transport via activating Wnt/ β-catenin pathway in the distal nephron and is derived from the cleavage by site-1 protease (S1P). The present study attempted to define the role of sPRR during salt-sensitive hypertension in SS rats. The effect of S1P inhibitor PF-429242 (PF at 20 mg/kg/d via mini-pump) on blood pressure, proteinuria, and urinary renin were tested in SS rats during high salt loading. In a separate group, a recombinant sPRR (sPRR-His at 20 μg/kg/d) was infused via catheterization of jugular vein to SS rats during PF treatment. In High sodium (HS)-loaded SS rats, administration of PF attenuated the hypertension development (MAP on day 10, HS+PF group: 124.5 ± 4.6 mmHg vs. HS group: 144.1 ± 8.2 mmHg) as assessed by radiotelemetry and proteinuria (HS+PF group: 50.9 ± 5.5 μg/24h vs. HS group: 105.3 ± 7.4 μg/24h). PF treatment reduced HS-stimulated urinary sPRR excretion by 65% in SS rats. Supplement of sPRR restored HS-induced hypertension during PF treatment (MAP on day 10, HS+PF+sPRR-His group: 138.7 ± 6.1 vs. HS+PF group: 124.5 ± 4.6 mmHg) and proteinuria (HS+PF+sPRR-His group: 79.0 ± 5.8 μg/24h vs. HS+PF group: 50.9 ± 5.5 μg/24h) in SS rats. Concurrently, PF treatment reduced HS-stimulated urinary renin activity (by 80%), aldosterone excretion (by 36%) and renal medullary prorenin and renin (by 60%), which were all restored by sPRR-His infusion (1.5- to 3-fold). Indices of Wnt/β-catenin pathway including protein levels of renal cytosolic Axin-2 and nuclear β-catenin as assessed by immunoblotting were elevated by HS (3.4-fold, 2.2-fold respectively) in SS but not SR rats, which were all suppressed by S1P inhibition (by 80% and 50% respectively) and restored by sPRR-His infusion. Given β-catenin signaling as a known regulator of intrarenal RAS, our results suggest that S1P-derived sPRR plays a pathogenic role in salt-sensitive hypertension in SS rats via β-catenin/intrarenal RAS pathway.
Abstract P221: The Mechanism of Salt-Sensitive Hypertension in Intermittent Hypoxia
Authors: Yeerbolati Alimila , Ogura Sayoko , Shimosawa Tatsuo , Maeda Hideyuki
Published: December 2018

The mechanism of Salt-sensitive Hypertension in Intermittent Hypoxia (IH)

Object:
The morbidity of sleep apnea syndrome (SAS) is about 4% of the general population and it has been recognized as an independent risk factor for hypertension as well as other cardiovascular diseases. We established IH condition for rats and mice to investigate the development of salt-sensitive hypertension under IH.

Methods:
FiO2 was adjusted to cyclically drop to 4-5% every 90 seconds for 8 hours. Male Sprague-Dawley rats at age of 11weeks old and C57B6j mice at age of 8 weeks were kept in IH for 2 weeks with normal salt (0.5%) and high salt (8% NaCl) diet. Blood pressure (BP) and heart rate (HR) were monitored by telemetry. To explore the mechanism of salt sensitivity, Na excretion was measured. At the end of study, we evaluated the activity of sodium-chloride cotransporter (NCC) and epithelial sodium channel (ENaC) by treatment with either hydrochlorothiazide (HCTZ) and Amiloride, respectively. mRNA of WNK4, ACE, ACE-2, Angiotensinogen, 11bHSD-2 were examined. In addition, we analyzed T cell population both by histologically and by FACS analyzer.

Results:
Only during IH, high salt diet-induced hypertension (+20-30mmHg) and BP decreased after changed to a low-salt diet. In normoxic controls, BP did not change by high salt diet. Natriuresis by HCTZ but not by amiloride was significantly enhanced IH group than normoxic control. There was no significant difference in mRNA level of WNK4, ACE, angiotensinogen, 11bHSD-2. CD3+ T cell was found around the renal tubule cells.

Conclusion:
The current study revealed that IH induces salt-sensitive hypertension and the enhanced Na+ re-absorption through the NCC may be involved. T-cell inflammation might be one of the reasons for activation of NCC.
Abstract P226: Reducing Sympathetic Activity and Cardiac Pre-Load Have Differential Effects on Mean and Pulsatile Blood Pressure Components
Authors: Faconti Luca , Farukh Bushra , Chowienczyk Phil
Published: December 2018

Objective:
Blood pressure (BP) can be divided into steady state (mean arterial pressure, MAP) and pulsatile (pulse pressure, PP) components that are differentially associated with target organ damage and cardiovascular events. Here, we tested the effects of non-pharmacological interventions that reduce sympathetic activity (SA) and cardiac pre-load on MAP and PP.

Material and Methods:
Pulse-wave analysis of the radial artery (SphygmoCor, AtCor Medical, Sydney, Australia), calibrated from 3 consecutive readings of brachial BP, was used for the evaluation of central PP and MAP. Measurements were obtained before and after supervised device-guided paced breathing (DGB), and bilateral lower limb venous occlusion (LVO, performed non-invasively using thigh cuffs), in random order. DGB reduces SA whereas LVO produces an acute reduction in pre-load. High resolution heart rate variability (HRV) as log-ratio of low-frequency/high-frequency range (LF/HF),( Schiller Medilog AR12plus, United States) was used to assess the effects of DGB on SA, while reduction of pre-load was evaluated by inferior vena cava diameter (IVCd) measured with cardiac ultrasound.

Results:
Thirty six HT patients (16 female) age (mean±SD) 46±12 years, BP 144.8±17.7/88.2±10.3 mmHg were studied. DGB decreased log LF/HF from 0.306±0.27 to 0.171±0.3 and LVO decreased IVCd from 15.16±3.96 mm to 12.39±3.43mm. Reduction of brachial BP was greater during DGB compared to LVO: (mean (95% confidence interval)):-9.7(-11.6,-7.8) mmHg vs -3.8(-5.8,-1.7) mmHg for systolic BP and -4.1(-5.6,-2.5) mmHg vs 1.1(-0.1, 2.2) mmHg for diastolic BP, respectively (
P<0.05)
. Effects of DGB and LVO were similar on PP: -3.5(-4.8,-2.3) mmHg for DBG and -3.1(-5.5,-0.7) mmHg for LVO, P>0.1. MAP reduction was significant only during DBG (-6.4 (-8.1,-4.7) mmHg) compared to LVO (-1.2 (-2.7, 0.1) mmHg,
P<0.01
between the interventions.

Conclusion:
Reducing sympathetic activity via DGB decreases both steady state and pulsatile components of BP whereas pre-load reduction selectively decreases PP. Depending on individual BP components, interventions targeting sympathetic activity and/or pre-load might be used to personalized treatment.
Abstract P231: Circulatory Gene Regulation in Youth With Primary Hypertension and Target Organ Damage
Authors: Mohammed Arif Fnu , Urbina Elaine , Sadayappan Sakthivel , Becker Richard
Published: December 2018

Introduction:
Primary hypertension (PH) is a multifactorial disease mainly influenced by genetic, epigenetic and environmental dynamics. Despite the occurrence of PH-associated cardiovascular (CV) events in youth, we have acquired only a limited understanding of epigenetic and gene regulation of blood pressure (BP)-related target organ damage (TOD) in adolescents.

Aims and Objectives:
To define circulatory gene and miRNA expression levels and allied signaling pathways in hypertensive adolescents (Hyp) with TOD.

Methods and Results:
Three hundred adolescents of both genders aged 11 to 18 years participated in the AHA-funded Study of Hypertension in Pediatrics - Adult Hypertension Onset in Youth (SHIP AHOY) Project. Mean clinic SBP was 104.3 ±8.1 and 133.0 ± 8.2 mmHg in normal
vs
Hyp (
p<0.005
) and mean daytime ambulatory SBP was 115.7 ±11.4 and 128.9 ±6.4 mmHg in normal
vs
Hyp (
p<0.005
), respectively. Youth with PH also had higher BMI (22.33 ±3.67
vs
31.56 ±9.42;
p<
0.05
), elevated serum creatinine (0.73 ± 0.18
vs
0.92 ±0.12;
p<
0.05
), and left ventricular hypertrophy (LVM/ht
2.7
; 25.4 ±1.8
vs
41.7 ±1.8;
p<
0.05
), and trend for increased arterial stiffness (PWV; 4.8 ±0.6
vs
5.8 ±2.0;
p
=0.18). mRNA and microRNA seq were performed using peripheral blood cells of 10 normal (mean 15.2 ±1.4 years; 50% male), and 10 Hyp patients (mean 15.5 ±1.5 years; 70% male). Seq data analysis revealed master genes (Fig. 1) and pathways that were differentially regulated in Hyp patients with TOD.

Conclusions:
PH in youth is associated with TOD that relates to a distinct gene expression profile. Knowledge of allied pathways may lead to new treatments in hypertensive youth to prevent future CV diseases.
Abstract P232: Hemodynamic Phenotype and Target Organ Damage in Adolescents With Spurious Hypertension
Authors: Obrycki Lukasz , Feber Janusz , Niemirska Anna , Litwin Mieczyslaw
Published: December 2018

Objective:
Spurious hypertension (sHTN) is characterized by elevated systolic blood pressure (BP) in peripheral arteries measured by office BP and ambulatory BP monitoring (ABPM), but normal central BP. The prevalence of sHTN and its clinical significance remains unknown. The aim of the study was to assess hemodynamics and target organ damage (TOD) in adolescents with sHTN in comparison with other forms of hypertension (HTN).

Patients and Methods:
We analyzed 265 children (54 girls; 15.1 ±2.5 years) referred with arterial HTN, in whom secondary HTN was excluded. We assessed BP levels - office BP, ABPM, central systolic BP (cSBP), hemodynamics - cardiac output (CO), stroke volume (SV) and target organ damage (TOD) - left ventricular mass index (LVMi), carotid intima-media thickness (cIMT), pulse wave velocity (PWV). Anova with TukeyHSD post hoc test was used for analysis.

Results:
115 subjects had white coat hypertension (WCH; office BP>95
th
perc+ABPM<95
th
perc), 25 had ambulatory prehypertension (preHTN; office BP<95
th
perc+ABPM<95
th
perc+ABPM load >25%), 43 had sHTN (office BP+ABPM >95th perc+cSBP<95
th
perc), 82 had true hypertension (tHTN; office BP+ABPM+cSBP >95th perc. There were no significant differences between groups in age, weight and BMI. Only pts with tHTN presented features typical of hyperkinetic circulation and pts with preHTN presented intermediate hemodynamic phenotype between WCH and sHTN and those with tHTN (p<0.001). Median LVMi ratio to upper limit of normal (g/m
2.7
/95
th
percentile by Khoury et. 2009) was similar in both WCH and sHTN (0.82 and 0.80, respectively), and significantly lower than in tHTN (0.89, p=0.04). The proportion of patients with LVH (LVMi>=95
th
perc) was 12% in WCH, 4% in preHTN, 9% in sHTN, 22% in tHTN (p=0.05). cIMT Z-scores were also significantly lower in both WCH (0.92) and sHTN (0.90) groups compared to tHTN (1.22, p=0.01). Similar results were obtained for PWV Z-scores (WCH=1.8, sHTN=1.29, tHTN=2.28, p=0.01). Patients with preHT were not significantly different from tHTN patients in all parameters.

Conclusions:
Adolescents with sHTN have similar hemodynamic phenotype and TOD as patients with WCH. However, because of lack of data on the evolution of sHTN, close monitoring and follow-up is warranted.
Abstract P234: Obstructive Sleep Apnea is More Common in Patients With Masked Uncontrolled Hypertension
Authors: Siddiqui Mohammed , Thomas Stephen J , Judd Eric K , Dudenbostel Tanja , Harding Susan , Oparil Suzanne , Calhoun David A
Published: December 2018

Introduction:
Masked uncontrolled hypertension (MUCH) is defined as controlled automated office blood pressure (AOBP) in clinic, but uncontrolled out-of-clinic BP by 24-hr ambulatory blood pressure monitoring (ABPM). Prior studies indicate that the prevalence of masked hypertension in patients with newly diagnosed obstructive sleep apnea (OSA) is nearly 30%. In addition, patients with OSA have a higher prevalence of masked hypertension than patients without OSA.

Aim:
Prospective determination of the prevalence and severity of OSA in patients with MUCH.

Methods:
In this prospective evaluation, 167 treated hypertensive patients were recruited from the University of Alabama at Birmingham Hypertension Clinic after having controlled BP at three or more clinic visits. All patients were evaluated by AOBP with the BpTRU device, ABPM, and diagnostic polysomnography (PSG) to determine the presence and severity of OSA based on the apnea-hypopnea index (AHI). Out of 153 who completed ABPM, 58 patients were controlled by AOBP and by ABPM, indicating true controlled hypertension and the remaining 95 patients were controlled by AOBP, but uncontrolled by ABPM, indicative of MUCH. 49 true controlled hypertensive and 69 MUCH patients completed PSG.

Results:
MUCH patients had a mean AHI of 10.3±13.3 compared to 4.5±10.2 events/hr in true controlled hypertension (p =0.045). Overall, the prevalence of OSA was 48.5% in patients with MUCH compared to 26.5% in true controlled hypertension (p = 0.01).

Conclusion:
MUCH has significant higher prevalence and severity of OSA compared to true controlled hypertension. These findings suggest that untreated OSA may contribute to the development of MUCH.
Abstract P254: Synergistic Effect of Angiotensin II and Amyloid-Beta on Brain Vascular Smooth Muscle Cell Senescence
Authors: Min Li-Juan , Bai Hui-Yu , Shan Bao-shuai , Kan-no Harumi , Higaki Akinori , Mogi Masaki , Iwanami Jun , Horiuchi Masatsugu
Published: December 2018

Objectives:
Cerebrovascular damage has been known to contribute to cognitive impairment and dementia. Amyloid-β (Aβ) induces cerebrovascular damage and is reported to stimulate endothelial cell senescence. We previously demonstrated that angiotensin II (Ang II) plays an important role in enhancing the vascular senescence. Here, we examined the possible cross-talk between Ang II and Aβ in regulating brain vascular smoot muscle cell (BVSMC) senescence.

Methods:
BVSMCs were prepared from adult male C57BL/6 mice by enzymatic digestion method. Cells were stimulated with Ang II with/without Aβ 1-40. Cellular senescence was evaluated by senescence associated β-galactosidase staining. Oxidative stress was determined by measuring superoxide anion production. Signal transduction was examined by Western blotting and luciferase activity assay.

Results:
Treatment with Ang II (100nM) or Aβ (1uM) at a relatively higher dose for 6 days significantly increased the percentage of senescent cells (25±1.43, 16±0.53 respectively) compared with control (13±2.22, 11±0.76 respectively). Treatment with Ang II less than 10nM or Aβ less than 0.5uM had no significant effect on cellular senescence. Interestingly, combination treatment of lower doses of Ang II (10nM) and Aβ (0.5uM) for 6 or 8 days markedly increased the percentage of senescent cells compared with control (control, 9±1.11; Ang II+Aβ, 15±0.77 for 6 days, control, 8±0.83; Ang II+Aβ, 15±1.14 for 8 days). Moreover, this lower dose combination of Ang II and Aβ induced significant increases in superoxide anion production, p-IκB, p16 and p53 expression, NF-κB activity for 4 days, p-ERK and p-IKKα/β expression for 2 days, and obvious decrease in pRb expression. These increases in senescent cells, superoxide anion production, expressions of p-ERK, p16, pRb by lower dose combination Ang II and Aβ were significantly inhibited by the treatment with U0126, an ERK inhibitor. However, U0126 had no significant effect on NF-κB activity.

Conclusion:
Ang II and Aβ synergistically promoted BVSMC senescence at least due to the enhancement of p-ERK-p16-pRb signaling pathway and oxidative stress, and the increase in NF-κB/IκB activity would be involved in this synergistic effect.
Abstract P256: Development of an Automated High Throughput Screen for GRK4 as Potential Novel Therapeutics for Treatment of Hypertension and/or Salt Sensitivity
Authors: Daley Michael G , Gildea John J , Xu Peng , Schiermeyer Katie A , Yue Wei , Marshall Stephen G , Jose Pedro A , Felder Robin A
Published: December 2018

G protein coupled kinase type 4 (GRK4) reduces renal sodium excretion by deactivating the renal dopamine type 1 receptor (D
1
R) through serine phosphorylation. At least 3 polymorphisms in GRK4 have been shown to be associated with the expression of hypertension and/or salt sensitivity in humans and in mice. GRK4 is an ideal therapeutic target for treating hypertension since putting human GRK4 variants in mice results in hypertension which can be reversed by reducing the expression of GRK4. Inhibitors of GRK4 could serve as potent and selective agents to promote sodium excretion in salt sensitive and/or hypertensive individuals. Therefore, we developed an automated high throughput homogeneous time resolved fluorescent resonance energy transfer assay (TR-FRET) to identify small molecule inhibitors of GRK4 isoenzymes. The assay relies on the close proximity (90 A) transfer of 337 nm energy from a europium anti-phosphoserine antibody (Eu-pSer) to an allophycocyanin labeled streptavidin acceptor (streptavidin-APC), producing FRET at 665 nm. The assay was optimized through serial dilutions of streptavidin-APC, Eu-pSer, GRK4, and a peptide consisting of serine sites in the D
1
R, serving as a substrate. To validate the assay and determine its suitability for a large scale high throughput screen, three inhibitors, GRKA, GRKB, and GRKC were selected and concentration response assays were performed to determine IC50 values and variability analysis. The IC50 and (R
2
) values of GRKA, GRKB, and GRKC were 4.99 μM (0.99), 26.52 μM (0.99), and 179.8 μM (0.82), respectively. GRKA is the most efficient GRK4 inhibitor, while GRKC is the least. This assay will be used to test congeners of compounds A, B, and C to identify more selective candidates, as well as for screening available small molecule compound libraries.
Abstract P257: Angiotensin Ii Type 1 Receptor Autoantibodies Decrease Angiotensin Ii Type 1 Receptor Internalization Through Attenuating The Recruitment Of β-arrestin1/2
Authors: Bian Jingwei , Zhang Suli , Lei Jinghui , Yin Xiaochen , Wang Pengli , Liu Huirong
Published: December 2018

Introduction:
Angiotensin II type 1 receptor autoantibody (AT1-AA) can continuously activate angiotensin II type 1 receptor (AT
1
R), which is related to cardiovascular disease, but the exact mechanism remains obscure. The activated AT
1
R signaling is adjusted or terminated effectively via the internalization, which is mediated by β-arrestin1/2 dependent-endocytic pathways. The hypothesis is that AT1-AA can attenuate β-arrestin1/2 recruitment, leading to the decreased AT
1
R internalization and the sustained AT
1
R activation.

Methods and Results:
Firstly, subcellular protein fractionation and Total Internal Reflection Fluorescence were used to examine the AT
1
R internalization induced by AT1-AA. The results confirmed that AT1-AA limited the AT
1
R internalization. Then, by co-expressing YFP-labeled AT
1
R and RFP-labeled β-arrestin1/2 in HEK293 cells with different stimulant for 10 min, the image showed that AT
1
R and β-arrestin1/2 were co-localized in the cytoplasm with Ang II. However, there were rare co-localization with AT1-AA. To further investigate the AT1-AA-induced recruiting of β-arrestin1/2, we recorded the relative change of BRET ratio of the interaction between YFP-labeled AT
1
R with Rluc-labeled β-arrestin1/2. The relative change of BRET ratio was significantly elevated with the time [(the BRET ratio with Ang II for 10 min were considered as 100%; 0 (β-arrestin1/2: 0.33 ±0.58/0.00 ±1.46), 2 (89.10 ±19.36/69.27 ±4.47), 5 (107.08 ±12.17/89.27 ±1.46), 10 (100.22 ±0.38/100.00 ±13.12), 20 (93.45 ±11.59/102.93 ±10.38) and 30 min (101.45 ±16.93/114.15±8.15)] and with the concentration (EC50
β-arrestin1/2
=6.69 ±4.09/6.34 ±2.29 nmol/L) after Ang II treatment. However, AT1-AA could not induce the recruitment of β-arrestin1/2. Furthermore, pre-incubation with an inhibitor of β-arrestin1/2 dependent-endocytic pathways prolonged the duration of AT
1
R downstream PKC, ERK1/2 phosphorylation, elevate intracellular Ca
2+
and vasoconstriction caused by Ang II, which simulated AT1-AA-caused persistent AT
1
R activation.

Conclusions:
Our data suggested that reduced AT
1
R internalization caused by AT1-AA was attributed to the inhibition of β-arrestin1/2 recruitment, which played a key role in how AT1-AA prolonged receptor activation.
Abstract P261: GPER is Required for Age-dependent Albuminuria and Glomerulosclerosis: Evidence for its Role in Podocyte Injury and Mesangial Nox1 Regulation
Authors: Meyer Matthias R , Daniel Christoph , Woods Christine D , Sharma Geetanjali , Fredette Natalie C , Amann Kerstin , Barton Matthias , Prossnitz Eric R
Published: December 2018

We recently found that the G-protein coupled estrogen receptor (GPER) exerts constitutive effects on cell proliferation and fibrosis in heart failure and arterial hypertension via the NADPH oxidase isoform Nox1 (Sci Signal 2016; 9(452): ra105). Whether GPER affects glomerulosclerosis or podocyte function is unknown. Thus, the present study investigated the effects of GPER in a model of age-dependent spontaneous focal segmental glomerulosclerosis and studied effects of GPER inhibition in mesangial cells and podocytes. Albuminuria and kidney histology were studied in male wild-type (WT) and GPER-deficient (

Gper
-/-

) mice at 4 and 24 months of age. Aged

Gper
-/-

mice were largely protected from albuminuria (albumin/creatinine-ratio, 0.9±0.4 vs. 3.5±1.0, -74 %, p<0.05 vs. WT).
Gper
deficiency had no effect at 4 months of age, but largely prevented age-dependent increases in kidney weight (306±11 vs. 554±94 mg), and glomerulosclerosis index (1.3±0.2 vs. 2.9±0.4, p<0.05 vs. WT). All changes were independent of blood pressure. In human podocytes exposed to TGFβ-1, treatment with the selective GPER blocker G36 markedly reduced mRNA expression of injury markers nephrin, collagen-4, and Wilms-tumor-1 (all p<0.01).
Gper
knock-down in rat mesangial cells reduced Nox1 protein expression by approx. 50% (p<0.05) while Nox2 and Nox4 expression remained unchanged. These results indicate that constitutive activity of
Gper
maintains Nox1 expression and contributes to podocyte injury in vitro and that
Gper
is essential for age-dependent podocyte injury, subsequent albuminuria, fibrosis and glomerulosclerosis in vivo. Nox1 downregulators such as G36 represent a new class of drugs that may offer therapeutic potential for patients with chronic renal diseases and other forms of chronic non-communicable diseases involving inflammation and fibrosis.
Abstract P263: Renal Proximal Tubule Cells from Inverse Salt Sensitive Individuals Have Lower Basal but Higher Sodium Stimulated ENaC Compared to Salt Resistant Individuals
Authors: Xu Peng , Gildea John J , Yue Wei , Jose Pedro A , Felder Robin A
Published: December 2018

Our previous studies on salt sensitivity has demonstrated that approximately 11% of study participants had a paradoxical increase in blood pressure (BP) (greater than 7-mm Hg) on a low NaCl diet and an increased basal renal proximal tubule cells (RPTC) Na
+
transport, which we define as inverse salt sensitivity (ISS). We hypothesized that the amiloride blockable Na
+
channel (ENaC) could be involved in ISS, particularly in the RPTC where up to 75% of renal nephron Na
+
reabsorption occurs. Previous investigators have localized αENaC in the distal nephron; however, we found αENaC is also present in the human RPTCs, as confirmed by tissue and cellular immunostaining, RT-PCR, and western blot with two selective αENaC antibodies. The presence of αENaC in human kidney tissue was confirmed by the Human Protein Atlas. Urine derived RPTCs from 3 ISS and 3 salt resistance (SR) clinical study participants were treated overnight with normal salt and high salt (140mM Na
+
and 170mM Na
+
respectively). High salt increased αENaC expression to a greater extent in ISS versus SR cells; however, ISS cells have significantly less αENaC at basal level (αENaC/actin: SR 1.41±0.248 VS ISS 0.592±0.157, n=3, P<0.05). Patch clamp ramp currents were recorded using a voltage ramp from -100 to +45 mV within 200 ms (N≥5 for each cell line). Na
+
current was measured at both 140 and 170 mM Na
+
with and without Bensamil (ENaC antagonist) and expressed as a ratio of current (170 mM/ 140 mM Na
+
). Bensamil blocked ENaC activity in ISS cells, but not in SR cells (Bensamil ratio/primary ratio: SR 1.149±0.026 VS ISS 0.818±0.017, n=3, t-test, P<0.001; final ratio/primary ratio: SR 1.088±0.022 VS 0.705±0.1, n=3, t-test, P<0.05). αENaC knockdown using siRNA significantly decreased Na
+
current in ISS cells (0.763±0.029, n=3, t-test, P<0.05) but not in SR cells (1.046±0.097, n=3, t-test, P<0.05). In conclusion, RPTC from ISS study participant urine demonstrated increased Bensamil Na
+
current compared to SR RPTC when exposed to increased sodium. This data could provide a novel functional assay for the diagnosis of ISS in the clinical setting and allow for an appropriate personalized medical intervention.
Abstract P266: Western Diet Impairs Small Vessel Relaxation and Initiates Kidney Endothelial Stiffening, Fibrosis and Tubulointerstitial Fibrosis Through the Endothelial Mineralocorticoidreceptor
Authors: Aroor Annayya , Habibi Javad , Ramirez-Perez Francisco I , Martinez-Lemus Luis , Jaffe Iris Z , Sowers James R , Jia Guanghong , Whaley-Connell Adam
Published: December 2018

Obesity enhances mineralocorticoid receptor (MR) activation, development of vascular stiffness and end organ injury. In this context, western diet (WD) activation of the endothelial mineralocorticoid receptor (ECMR) contributes to endothelial cell stiffening and promotes maladaptive inflammatory responses and fibrosis in cardiovascular tissue of female mice. However, the role of ECMR on kidney endothelial stiffening, inflammation and fibrosis remains unknown. We hypothesized that deletion of the ECMR would prevent WD-induced increases in endothelial cell stiffness, reductions in bioavailable nitric oxide (NO), increased perivascular and tubulointerstitial inflammation oxidant stress, and fibrosis in females. Four-week-old female ECMR knockout and wild-type mice were fed either a mouse chow or a WD high in saturated fat and refined carbohydrates for 16 weeks. Without blood pressure changes between groups, WD-feeding increased body weight and fat mass as well as indices of vascular stiffness (pulse wave velocity and kidney endothelial cell stiffness) and impaired endothelial-dependent vasodilatation. The WD-induced kidney endothelial cell stiffness was associated with attenuated endothelial NO synthase activation, increased oxidative stress, along with pro-inflammatory immune responses, alterations in extracellular matrix degradation pathways and tubulointerstitial fibrosis. ECMR deletion prevented these abnormalities through improvements in endothelial NO synthase and reductions in macrophage polarization, LARP6, TG2 and MMP2. Our data support that activation of ECMR contributes to endothelial dysfunction, increased permeability and stiffening in the kidney which, in turn, promotes macrophage infiltration, M1 polarization, inflammation and oxidative stress, resulting in alterations in matrix degradation that promote tubulointerstitial fibrosis in females consuming a WD.
Abstract P270: Increase of Total Peripheral Vascular Resistance and Arterial Stiffness in Non-Diabetic Hypertensive Patients With Overweight, Compared With Hypertensive Patients of the Same Age With Normal Body Mass Index
Authors: Cabrera-Sole Ricardo , Turpin Lucas Caridad , Garcia Ruiz Santiago , Urrego Rivera Liliana , Stephan Luekpe Erik , Aguilera Saldaña Manuel
Published: December 2018

Introduction:
it is known that obese hypertensive patients (HTAO) have more difficulties for the better control of blood pressure, being involved many factors that try to explain this situation. However, little has been said regarding total vascular resistance and arterial stiffness (AS) in overweight patients (OWHTA), so we present this work that could provide data of interest in these

Objectives:
to assess the relationship between adequate control of blood pressure, AS and TPR in patients with OWHTA compared with hypertensive patients with a normal body mass index (BMI).

Material and Method:
cross-sectional and prospective study of a cohort of 1700 male patients (P) with hypertension, we have 290 P divided into two groups: OWHTA Group 190 P with BMI between 25 and 29 (65 ± 8 years) and Group HTA: 100 P with BMI <25 (66 ± 6 years) who have received treatment from their GP, to keep the BP below 140/80 and apparently were well controlled. A 24 hours blood pressure holter (ABPM) was performed in all patients, and augmentation index (AI) ,pulse wave velocity (PWV) as well as the TPR were measured. The results were compared in both groups and are presented in the following table:

Results:
Table of results:

*means p value less than 0.05.

Conclusions:
according to our data, in hypertensive males with overweight there are already aggravating factors such as the difficulty for better hypertension control , increase in AS and a significant increase in the TPR that has been seen in obese patients. All of these factors make the cardiovascular risk of these patients much greater, and forces the physician to be more demanding with the therapeutic measures.
Abstract P272: Gender Features of Angiological Screening in Young People
Authors: Evsevyeva Maria , Eremin Mikhail , Rusidi Anjelika , Fursova Elena , Koshel Vladimir , Baturin Vladimir
Published: December 2018

Relevancy:
The wide occurrence of risk factors (RF) necessitates rapid introduction of angiological screening.

Purpose:
To determine the results of mass instrumental evaluation of vascular stiffness in students taking into account gender and cases of EVA-syndrome.

Methods and Materials:
It was surveyed 224 students on base of Student Health Centre of StSMU as a part of project “ The University of healthy life style” for study of FR profile, including fat metabolism by using rapid diagnosis "Lipid Panel" (USA). It was valued the condition of the vessel wall according to Cardio-Ankle Vascular Index (CAVI) from left (L) and right (R) sides thanks to device Vasera VS-1500(‘Fukuda Danshi’, Japan). The device also gives an opinion on vascular age, which allows to identify cases of early vascular aging or EVA syndrome. Statistics analyses were made with programme ‘ Statistica 10.0’( StatSoft Inc, USA).

Results:
Among boys in the upper quartile of CAVI-R and CAVI-L there were 18 (25,4 %) and 17 (23,9%) , but among the female - 30 (19,6%) and 26 (17,0%). Signs of syndrome EVA also were recorded more often in boys than in girls - 9 (12,7 %) versus 12 (7,8%). Noteworthy that there were some differences in lipidogram in carriers of this syndrome, depending on sex. Thus, girls' total cholesterol and low density lipoprotein levels were 4.4±0.34 and 2.76±0.24 versus 3.7±0.11 and 2.3±0.32 mmol/l in males. But at same time, high density lipoproteins were also higher in the fairer sex - 1,3±1,2 against 0,98±0,06 mmol/l in boys. In girls triglycerides were registered at lower level - 0,72±0,1 against 0,98±0,25 mol/l. However, all described differences did not reach a reliable level. The average value of CAVI-R and CAVI-L for male were 5,80 ± 0,09 and 6,01 ± 0.09, but for female - 5,58 ± 0,06 and 5,90 ± 0,05. The median of CAVI-R and CAVI-L for men was 5, 81 and 6,02, but for the women - 5,73 and 5,81.

Conclusion:
In upper quartile of CAVI values boys fall almost a third more than girls. Carriers of EVA syndrome are also almost half the most common among representatives of the stronger sex. At the same time, not all indicators of lipid status in boys with EVA syndrome are worse than in their girl-peers with same syndrome. These data can be useful in forming groups of increased cardiovascular risk among young people.
Abstract P279: Regulation Of H
2
S Production By Cystathionine Gamma-lyase In Endothelial Cells.
Authors: Mendiola Perenkita J , Gonzalez Bosc Laura V , Naik Jay , Kanagy Nancy L
Published: December 2018

Hydrogen sulfide (H
2
S) is the most recently described endothelium-derived vasodilator. In the endothelium, H
2
S is predominantly produced by cystathionine γ-lyase (CSE). We and others have previously shown CSE inhibition diminishes acetylcholine (ACh)-induced dilation in aortic ring segments. However, the mechanism of CSE regulation is not well-defined. The goal of this study was to identify endogenous regulators of CSE in endothelial cells. Previous reports suggest that [Ca
2+
] and phosphorylation regulate CSE activity. We previously reported that there was an increase in H
2
S production in rat aortic endothelial cells loaded with an H
2
S fluorescence indicator, sulfide fluor-7 acetoxymethylester (SF7-AM, 10 μM) after exposure to either ACh (10 μM) or an H
2
S donor (NaHS, 10 μM). Pretreatment with the CSE inhibitor, β-cyanoalanine (BCA, 100 μM) prevented the ACh-induced but not the NaHS-induced increases in SF
7
flourescence. Our current study tested the hypothesis that increasing intracellular calcium stimulates CSE-dependent H
2
S production. Cultured rat aortic endothelial cells were loaded with SF
7
and a nuclear dye (Nuclear-ID Red DNA stain, to correct for cell number) and fluorescence was measured at baseline and in response to a calcium ionophore (ionomycin, 100 nM). Ionomycin increased H
2
S production (Control: 0.8293, +ionomycin: 1.0365, p = 0.0001, n=3 of 7-15 wells per replicate). Pretreatment with BCA significantly decreased ionomycin-induced H
2
S production (+ionomycin: 1.0365, +BCA: 0.6600, p < 0.0001, n=3 of 7-15 wells per replicate). Preliminary ongoing studies in pressurized mesenteric arteries suggest a tendency of CSE inhibition to reduce ACh-induced dilation (Control: 77.06%, +BCA: 42.04%, p=0.08, n=3-4 arteries per group). However, a larger sample size is needed. Taken together, these studies suggest that ACh elicits vasodilation through a Ca
2+
-dependent activation of CSE.