bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 289: Reduced Uterine Perfusion Pressure (RUPP) Elicits Increased sFlt-1 Levels Not Only In The Placenta But Also Adipose Tissue
Authors: Spradley Frank T , Palei Ana C , Granger Joey P
Published: March 2022

Preeclampsia is a pregnancy-specific disorder characterized by new-onset hypertension. Placental ischemia is causative in this disorder with the release of antiangiogenic factors such as sFlt-1 into the maternal circulation promoting endothelial dysfunction and hypertension. Obesity is a major risk factor for preeclampsia, but the mechanisms are far less understood. Recent evidence suggests that adipose tissue is a source of sFlt-1. This study tested the hypothesis that placental ischemia produced by RUPP elicits an increase of sFlt-1 levels in both placental and adipose tissue from normal pregnant rats. Timed-pregnant Wistar-hannover rats (20 wks old) on NIH31 standard chow were subjected to RUPP (N=15) on gestational day (GD)14 or remained normal pregnant (NP, N=14). Rats were implanted with carotid catheters on GD18 and fasted overnight. On GD19, mean arterial blood pressure (MAP) was assessed in conscious, restrained rats. Statistical significance was P<0.05. MAP was greater in RUPP (114±2 v 101±1mmHg) with reduced fetal weight (1.73±0.02 v 1.88±0.01g) but similar placental weight (RUPP: 0.45±0.03 v NP: 0.47±0.03g). Although RUPP reduced body weight (297±7 v 343±6g), visceral adipose tissue weight was not altered (RUPP: 11.5±1 v NP: 13.2±1g). Total cholesterol was increased (RUPP: 223±35 v NP: 156±6mg/dL) but there was no difference in free fatty acids (RUPP: 8±2 v NP: 8±2mg/L) but reduced triglyceride levels (RUPP: 265±38 v RUPP: 659±100mg/dL). RUPP reduced leptin (3.2±0.2 v 4.2±0.4ng/mL) and adiponectin (2.8±0.2 v 3.3±0.2ug/mL) with increased fasting glucose levels (191±6 v 163±8mg/dL). Very interestingly, RUPP increased sFlt-1 levels in placenta (4702±375 v 3903±309pg/g tissue) and retroperitoneal adipose tissue (179±28 v 76±22pg/g tissue). These data indicate that RUPP-induced hypertension in normal pregnant rats promotes metabolic disturbances along with increases in sFlt-1 not only the placenta but in the adipose tissue. In conclusion, we propose that placental ischemia-induced hypertension is exaggerated in states linked to increased accumulation of adipose tissue, such as in diet-induced or genetic obesity, due to amplified metabolic derangements and increases in sFlt-1 levels from both placental and adipose sources.
Abstract 314: Evidence for the Expression of Renin Angiotensin System (RAS) and a Disintegrin and Metalloproteinase (ADAM) 17-mediated shedding of ACE2 in COS-7 cells
Authors: Kashkari Nada , Grobe Nadja , Chodavarapu Harshita , Somineni Hari K , Singh Richa , Di Fulvio Mauricio , Elased Khalid M
Published: March 2022

The renin angiotensin system (RAS) plays a vital role in the regulation of the cardiovascular and renal system. COS-7 is a robust and easily transfectable cell line derived from the kidney of the African green monkey, Cercopithecus aethiops. The aim of this study was to 1) demonstrate the presence of an endogenous and functional RAS in COS7 cells and 2) to investigate the role of ADAM17 in the ectodomain shedding of ACE2. COS-7 cells were grown to 90% confluency followed by incubation in serum-free media for 24 hours. Western blot, immunohistochemistry and mass spectrometry (MS)-based enzyme assays were used to study RAS protein expression and activity in COS7 cells. Western blot and immunostaining confirmed endogenous expression of ACE (195 kDa), ACE2 (75-70 kDa), AT1R (43 kDa), renin (41 kDa) and ADAM17 (130 kDa) in COS7 cells. A sensitive and selective MS approach was used to determine endogenous RAS enzymatic activity in COS-7 lysate and media using the natural RAS substrate Ang II (m/z 1046). MS analysis detected Ang-(1-7) formation (m/z 899) in lysate and media of COS7 cells. While Ang-(1-7) formation in media was completely blocked by the ACE2-specific inhibitor MLN-4760, Ang-(1-7) formation in the lysate was significantly inhibited by the prolyl carboxypeptidase-specific inhibitor Z-pro-prolinal. Using short-hairpin (sh) RNA-mediated technology, ADAM17 protein expression and activity was significantly reduced in silenced cells compared to normal cells. Western blot analysis showed a significant increase of ACE2 protein expression in the lysate of silenced cells compared to normal cell and ACE2 shedding into the media was significantly reduced in silenced cells. This is the first study to demonstrate endogenous expression of the RAS and ADAM17 in COS-7 cells. Use of COS-7 could enhance our understanding of the cross talk between different arms of the RAS and dissect the direct effects of ACE inhibitors and Ang II receptor blockers. The transfectable nature of this cell line makes it an attractive in vitro cell model for studying the molecular, functional and pharmacological properties of the renal RAS. The data support the utility of COS-7 for the study of ectodomain shedding of ACE2 in vitro.
Abstract 411: Diurnal Variation of Blood Pressure in Medical Residents: MARINE 1 (Measures of Active Residents in Numerous Environments)
Authors: Brown Lorrel , Chaisson Jordan , Cheng Henry , Labaki Wassim , Cingolani Oscar , Schulman Steve , Desai Sanjay
Published: March 2022

Residency training subjects healthy young people to frequent fluctuation in sleep-wake cycles. Although shift work is known to increase cardiovascular risk, little is known about the physiology of phase-shifted sleep in residency. This study uses 24-hr ambulatory blood pressure monitoring to investigate diurnal BP variation in medical residents working alternating day/night periods. We hypothesized that diurnal variation of BP is dependent upon sleep/wake status, not upon day/night hours.

Residents were recruited from the Johns Hopkins Medicine Residency Program during the 2013-2014 academic year. Thirty-six individuals participated while working in three clinical environments (intensive care unit, general medicine wards, outpatient clinic), and during various sleep-wake schedules (night shift, 24-hr overnight call, day shift). BP was measured every 30 min during waking hours and every 2 hr during sleep. Residents self-reported during which hours they slept. Ninety-eight separate 24-hr periods of data were collected (Table 1).

There is no difference in
mean
24-hr SBP,
awake
SBP/DBP, or
asleep
SBP/DBP between those working different schedules. However, there are statistically significant differences in
day
SBP/DBP and
night
SBP/DBP between participants working different schedules. In all schedules, participants showed at least a 9 mmHg drop in SBP between
awake
and
asleep
. Diurnal variation of BP in this population is present, and dependent upon sleep/wake status, not day/night hours. Further investigation into the correlation between this reversal of diurnal BP variation and cardiovascular outcomes in shift workers is warranted.
Abstract 427: Carotid Disease in Postmenopausal Women Correlates with Cardiac Hypertrophy and Ageing
Authors: Pereira Valter L , Franqueiro Luiz F , Campos Luciana A , Baltatu Ovidiu C
Published: March 2022

Background:
The atherosclerotic carotid disease is characterized by thickening of the intima-media of the vascular wall (IMT), which is considered as a reliable predictor of cardiovascular events. The aim of this study was to investigate a relationship between IMT and nonatherosclerotic cardiac and vascular risk factors in postmenopausal women.

Methods:
Left and right carotid IMT, plaque score and resistive index (common, external and internal carotid RI), left ventricular (LV) wall thickness and ascending aorta diameter were determined by echography.

Results:
Women’s and men’s age was normally distributed with mean(SD) of 71(11) and 68(11), respectively. Left and right carotid IMTs were strongly correlated to each other (Spearman's r = 0.71 in women and 0.89 in men, p < 0.001). Only the right IMT was correlated with the plaque score at both sexes (Spearman's r = 0.36 in women and 0.37 in men, p = 0.01). The carotid resistive indexes had no significant relationship with the C-IMTs. The LV wall thickness and the ascending aorta diameter had a significantly positive correlation with the right IMT in women (Spearman's r = 0.31 and 0.31, p < 0.05) but not in men. The carotid RIs had a significantly positive association with age in both sexes (Spearman's r ranging between 0.30 and 0.56, p < 0.05). However, right IMT, LV thickness and aortic diameter were positively correlated with age only in women (Spearman's r = 0.33, 0.40 and 0.26 respectively, p < 0.05).

Conclusion:
These data demonstrate that nonatherosclerotic risk factors are associated with higher IMT in women but not in men. The synergic action of these factors may contribute to a worse carotid deterioration in postmenopausal women.
Abstract 435: The Association Between Initial Antihypertensive Prescription Interval And Medication Adherence: A Cohort Study Among 203,259 Patients
Authors: Wong Martin C , Tam Wilson W , Wang Harry H , Chan Wai Man , Kwan Mandy W , Cheung Clement S , Tong Ellen L , Cheung Ngai Tseung
Published: March 2022

Background:
Optimal adherence with antihypertensive medications is crucial to prevent hypertension-related complications. This study evaluated whether the interval of initial antihypertensive prescriptions is associated with better medication adherence among hypertensive patients who received their first-ever antihypertensive agent.

Methods:
From a validated clinical database of all patients in the public healthcare sector in Hong Kong, we included all patients who were newly prescribed an antihypertensive agent from 2001 to 2005. They were followed-up for 12 months, and optimal adherence was defined as having the Proportion of Days Covered (PDCs) ≥ 0.80. A binary logistic regression analysis was conducted to test the association between initial prescription interval and adherence level in the subsequent 12 months.

Results:
From 203,259 eligible patients with an average age of 58.7 years (SD 17.3), the overall adherence rate was 32.4% [65,856/203,259]. The proportions of patients whose initial prescriptions lasted for ≤ 6 days; 7-14 days; 15-28 days and ≥ 29 days were 23.7%, 24.3%, 15.1% and 37.0%, respectively. After controlling for patients’ age, sex, socioeconomic status, service type, antihypertensive drug class, and district of residence, those whose initial prescription was 15-28 days (adjusted odds ratio [AOR]: 1.90, 95% C.I. 1.82-1.99, p<0.001) and ≥ 29 days (AOR=4.13, 95% C.I. 3.96-4.31, p<0.001) were significantly more likely to be adherent than those who were prescribed for ≤ 6 days.

Conclusions:
These findings showed that short initial prescription interval was associated with poorer medication adherence, and this practice should be cautious.
Abstract 437: Racial/Ethnic Differences in Cardiovascular Disease Outcomes in Hypertensive Patients
Authors: Pu Jia , Zhao Beinan , Chung Sukyung , Nimbal Vani , Wang Elsie , Fortmann Stephen , Palaniappan Latha
Published: March 2022

Background:
Higher rates of CVD have been found in Mexicans and some Asian American subgroups as well as African Americans. However, little is known about the association between HTN, a main risk factor of CVD, and CVD among rapidly growing minority groups. Our study aims to identify racial/ethnic differences in CVD outcomes in HTN patients across Asian American subgroups (Asian Indian, Chinese, Filipino, Japanese, Korean, Vietnamese), Mexican, African American, and non-Hispanic White (NHW) in a large, mixed payer ambulatory care setting in northern California.

Study Design:
We examined electronic health records of patients age 18 years or older with at least one primary care visit from 2008-2012. CVD was defined by ICD-9 codes while HTN was defined by ICD-9 codes, antihypertensive medication, or two or more abnormal blood pressure readings (>140/90mm Hg). We estimated the rate of CVD incidence among HTN patients. Follow-up period was six years on average.

Results:
African Americans had the highest prevalence of HTN (51%, 2,715 out of 5,328), followed by NHWs (35%, 96,965 out of 276,133), Mexicans (34%, 3,867 out of 11,532), and Asian Americans (24%, 29,872 out of 124,139). Among these HTN patients, 9% (11,667 out of 133,419) developed CVD by the end of year 2012. HTN patients with CVD were more likely to be older, male, smoker and have a higher BMI. There was no significant difference in baseline BP levels. Compared to NHWs, both African American (OR=1.84, CI: 1.61-2.10) and Mexican HTN patients (OR=1.28, CI: 1.12-1.46) were at a higher risk of developing CVD after adjusting for age and gender, while Asian Americans were at a lower risk (OR: 0.81, CI: 0.79-0.86). Racial/ethnic differences also presented across the Asian American subgroups with Filipino and Asian Indian HTN patients at an elevated risk of developing CVD compared to other Asian American subgroups.

Conclusions:
Among HTN patients, there is heterogeneity across R/E minority groups in their risk of developing CVD. It might be differential utilization pattern such that Mexicans, Filipinos, and Asian Indian are likely to receive HTN treatment at more severe stage. Further studies are needed to better understand the pathways of CVD development among HTN patients of different race/ethnicity.
Abstract 440: Gender Differences In Hemodynamic And Autonomic Reactivity During Mental And Physical Laboratory Stress Tests In Normal Subjects Of The “Oman Family Study”
Authors: Hassan Mohammed O , Jaju Deepali S , Bayoumi Riad A , Al-Yahyaee Saeed , Comuzzie Anthony G , Voruganti Saroja V , Albarwani Sulayma A
Published: March 2022

Background:
Exaggerated cardiovascular reactivity for blood pressure (BP) and heart rate (HR) to laboratory stress tests has been proposed as a predictor of cardiovascular disease.

Subjects and Methods:
The study was conducted in normal subject; 262 males and 280 females (mean age 30 ± 9 years, BMI 23.5 ± 4 years), from five large and highly consanguineous homogeneous Arab families of OFS. Hemodynamic parameters: Heart rate (HR), systolic, diastolic, mean BP (mmHg), Stroke index (SI, SI (ml/beat/m
2
), cardiac index (CI,L/min/m
2
) ), total peripheral resistance index (TPRI, yne*s*/cm
2
)), left ventricular ejection time (LVET, ms) and cardiac contractility (IC, 1000/sec ) were computed from beat-to-beat BP, ECG and impedance cardiography (TFM, CNSystems). Autonomic parameters ( LF; sympathetic , HF; parasympathetic, normalized) and baroreceptor sensitivity (BRS, ms/mmHg) were computed using spectral analysis and the sequence method, respectively. Hemodynamic and autonomic eactivity were averaged after 3 minutes of the word conflict (Δ WCT) and the cold presser (Δ CPT) tests. Results were compared using the Student’s paired ‘t’ test. A P value of < 0.05 was considered as significant.

Results:
Males had significantly higher or lower reactivity than females for ΔWCT HR( 9.3 ±7 vs. 6.8 ± 6.3), SBP (9.9 ± 11 vs. 7.5 ±6.) , DBP (7.9 ± 8 vs. 6.3± 6), MBP (9.4 ±9.5, (SI -1.9 ± 5.5 vs. - 0.8 ± 4.7) and BRS ( -7.5± 15.9 vs. -4 ±10.5) . In contrast ΔCPT in females evoked significantly higher HR (8.2 ± 8 vs. 5.1 ± 8), CI (0.19 ± 0.4 vs. 0.08 ±0.4) and LF (11.6 ± 16.vs. 5.8 ±15.9) but significantly lower LVET (-8.7 ±16.6 vs. -3.8 ±9) and , BRS ( -6 ± 13 vs.-3.4 ±14 )

Conclusion:
In this genetically and environmentally homogeneous cohort of OFS we found significant gender differences in hemodynamic and autonomic responses during the WCT and CPT tests. This may indicate that biological mechanism and genetic factors may contribute to such differences and should be receive special emphasis in cardiovascular research.
Abstract 462: Effect of Adding Beet Root Juice to Exercise on Physical Performance and Cardiovascular Measures in Old Hypertensive Subjects
Authors: Shaltout Hossam A , Marsh Anthony P , Laurienti Paul , Basu Swati , Klebous Cassandra C , Kus Nicole , Morgan Ashley , DosSantos Patricia C , Norris James L , Miller Gary , Rejeski Jack , Hawfield Amret T , Diz Debra I , Kim-Shapiro Daniel B
Published: March 2022

Hypertension and diminished blood flow to muscles and brain are common problems for the elderly that may lead to impaired cognition and physical performance and poor cardiovascular outcomes. Exercise improves blood pressure and brain blood flow. Thus, any intervention that can improve exercise effects in hypertensive individuals may have beneficial effects. High nitrate containing beetroot juice (BRJ) has been demonstrated to increase exercise tolerance and improve blood flow to working muscles. In this study we examined the effect of adding BRJ to an exercise regimen on exercise performance and cardiovascular measures in old subjects. Participant (n=26; mean age=65 yrs, range 57-77) with controlled hypertension (on 2 or more medications) were assigned to either exercise with BRJ+nitrate (BRJ+) (n=13, 6F) or BRJ-nitrate placebo drink (Placebo) (n=13, 7F). The exercise intervention was a 6-week treadmill walking program of self-paced. Participants walked 3 times per week for a maximum of 50 minutes per session. Participants also consumed 140 ml BRJ+ nitrate or BRJ-nitrate daily for 6 weeks. There were no differences between groups in mean VO
2
max at the beginning of study (placebo = 19.4 ± 0.5 ml O
2
/kg/min; BRJ = 19.4 ± 0.5 ml O
2
/kg/min) and change VO
2
max by the end of the intervention (increase of 1.15 ml O
2
/kg/min in placebo vs 0.9 ml O
2
/kg/min in BRJ+). There were no differences between groups in 24 hour ambulatory mean arterial pressure (MAP) or heart rate at the beginning of study. Heart rate and MAP showed similar reductions in both groups (4 beats/min for heart rate and 5 mm Hg for MAP). Similarly total arterial compliance was improved in both groups (placebo from 15.1 ± 3.7 ml/mm Hg to 18.3 ± 4.8, p = 0.02; BRJ+ from 18.3 ± 5.0 to 20.5 ± 3.9 ml/mm Hg; p = 0.04). There was no additional benefit of BRJ+ in the cardiovascular measures above that seen with exercise alone.

In summary, in this cohort of controlled hypertensive elderly, addition of BRJ with nitrate to exercise has no additional benefits on physical performance or cardiovascular function. Further studies are warranted in different cohorts such as drug naïve or uncontrolled hypertensive to further characterize any possible benefits of BRJ. Support from WFU Translational Science Center
Abstract 467: A Combination of Metabolic Syndrome Components Increased the Risk of Incident Atrial Fibrillation in a General Urban Cohort: the Suita Study
Authors: Kokubo Yoshihiro , Kobayashi Takashi , Watanbe Makoto , Kamakura Shiro , Kusano Kengo , Kawanishi Katsuyuki , Miyamoto Yoshihiro
Published: March 2022

Background:
Atrial fibrillation (AF), the most common chronic arrhythmia, is linked to increased risks of mortality and cardiovascular disease. Few prospective studies have examined the combined effect of metabolic syndrome (Mets) components on the incidence of AF in a general population. We investigated whether combinational Mets components increase the risk of incident AF in an urban Japanese population.

Methods:
The 6,659 Suita Study participants (30-84 yrs old) were initially free of AF and prospectively followed up for incident AF. Standard 12-lead ECGs were obtained from all subjects in the supine position. The participants were diagnosed with AF if AF or atrial flutter was present on ECGs obtained during a biannual routine health examination or if AF was indicated as a present illness by questionnaire, medical records, or the registration of one or more cardiovascular disease events during follow-up. The Mets components were defined as follows: higher blood pressure (HBP, systolic/diastolic BP ≥130/85 mmHg or use of antihypertensive medication), hyperglycemia (fasting plasma glucose >100 mg/dL or use of diabetes medication), hypertriglyceridemia (HTG, serum triglyceride >150 mg/dL or use of medication for HTG), lowered HDL cholesterolemia (HDL <40 mg/dL in men and <50 mg/dL in women), and central obesity (waist circumference >85 cm in men and >80 cm in women). The definition of Mets was used for the current international recommendations, which constitutes the presence of any three or more of five components. Cox proportional hazard ratios (HRs) and 95% confidence intervals (CIs) were analyzed after adjusting for age, gender, smoking, drinking, chronic kidney disease, stroke, and heart disease at baseline.

Results:
During the 13 years of follow-up, 242 incident AF events occurred. The adjusted HRs (95% CIs) of incident AF for central obesity, HBP, and Mets were 1.39 (1.06-1.82), 1.41 (1.08-1.85), and 1.34 (1.03-1.75), respectively. Compared with the absence of the components, the adjusted HR (95% CIs) of incident AF for subjects with any three components was 1.72 (1.11-2.67).

Conclusion:
Mets, central obesity, and HBP are risk factors for incident AF. Appropriate BP and body weight are important for preventing AF in the general population in Japan.
Abstract 472: Positive Role of Combined Exercise Training in a Model of Metabolic Syndrome and Menopause: Autonomic, Inflammatory and Oxidative Stress Evaluations
Authors: Conti Filipe F , Brito Janaina O , Dias Danielle S , Bernardes Nathalia , Malfitano Christiane , Sanches Iris C , llesuy Susana , Irigoyen Maria-Claudia , De Angelis Kátia
Published: March 2022

The aim of this study was to evaluate the effects of combined exercise training in hypertensive ovariectomized rats submitted to fructose overload. Wistar and spontaneously hypertensive (SHR) rats were divided into (n=8/group): control (C), hypertensive (H), hypertensive ovariectomized (HO), hypertensive ovariectomized submitted to fructose overload sedentary (HOF) and trained (HOFT). The combined exercise training was performed on a treadmill and ladder adapted to rats in alternated days, during 8 weeks. At the end of exercise protocol, arterial pressure (AP) signals were directly recorded. Cardiovascular autonomic modulation was evaluated by spectral analysis. Oxidative stress and inflammation were measured on cardiac tissue. The association of hypertension and ovariecyomy induced an increase in arterial pressure (AP) (HO: 164±5 vs. H: 146±6 mmHg) and body weight in relation to hypertensive rats. The association of risk factors (hypertension+ovariectomy+fructose) induced an additional increase in AP (HOF: 174±4 mmHg), increased heart rate (HOF: 403±12 vs. HOS: 348±16 bpm) and blood triglycerides (HOF: 160±8 vs. HO: 125±6.4 mg/dl), promoted impairment on the insulin sensitivity, cardiovascular autonomic modulation, oxidative stress (redox ratio of glutathione: HOF: 8.94±0.8 vs. HO: 13.00±1.4) and inflammation (TNF-α- HFO: 65.8±9.9 vs. HO: 31.7±8.6 pg/mg protein) of these animals. However, the combined exercise training was able to reduce the AP (HOFTc: 158±4 mmHg) and heart rate (HOFT: 303±5 bp), to normalize the triglyceride levels (HOFT: 137±4.7 mg/dL), insulin sensitivity and vascular autonomic modulation. Moreover, this approach reduced cardiac lipoperoxidation and increased antioxidant defense and redox balance of glutathione (HOFT: 11.94 ± 1.1), reduced TNF-α (HOFT: 33.1±4.9 mg/dL) and increased IL-10. The results of this study demonstrated that the association of risk factors promoted additional impairment in metabolic, cardiovascular, autonomic, inflammatory and oxidative stress parameters and the combined exercise training was able to reduce and/or normalize these dysfunctions.
Abstract 478: Development Of A Novel Technique To Visualize In Vivo Renin Activity And Assess Its Use In The Study Of The Pathogenesis Of Diabetic Nephropathy
Authors: Satoh Minoru , Kidokoro Kengo , Kashihara Naoki
Published: March 2022

Introduction and aims:
Although systemic renin activity is suppressed in diabetes, a renin-angiotensin system (RAS) inhibitor exerts renoprotective effects. Activation of local/tissue RAS, therefore, appears to be involved in the pathogenesis of diabetic nephropathy. We attempted to develop a technique to visualize tissue renin activity in vivo using the fluorescence resonance energy transfer (FRET) system and multiphoton laser microscopy (MP-LM). In the FRET peptide, the fluorescence of 5-fluorescein amidite (FAM) is intramolecularly quenched. After cleavage into two fragments by renin, the fluorescence of 5-FAM is recovered and can be visualized at excitation/emission wavelengths of 490/520 nm.

Methods and results:
Diabetes was induced with streptozotocin (STZ) in C57B6/ mice, followed by aliskiren (Alis: 25 mg/kg/day with osmotic pump) or valsartan (Val: 15 mg/kg/day by lavage) treatment for 4 weeks starting at 6 weeks after STZ injection. Diabetic mice (DM) showed higher urinary albumin excretion (UAE; control: 85±15 μg/day, DM: 220±25 μg/day, P<0.05). Alis or Val treatment significantly reduced UAE (Alis: 120±21 μg/day, Val: 135±18 μg/day). Renin activity in the kidney assessed by the FRET system was markedly enhanced in the glomeruli in DM (1.7±0.2-fold that in controls, P<0.05). Alis reduced plasma and renal renin activity to baseline levels. Production of reactive oxygen species (ROS) and nitric oxide (NO) was evaluated by MP-LM using dichlorofluorescin and diaminorhodamine-4M acetoxymethyl ester. Increased ROS production and diminished bioavailable NO were noted in the glomeruli in DM, but were significantly ameliorated with Alis or Val treatment. Glomerular permeability of macromolecules was visualized in vivo using MP-LM with 70-kDa dextran. Glomerular permeability was significantly greater in DM than in the controls and was ameliorated by Alis or Val treatment.

Conclusions:
We developed an in vivo imaging technique to visualize renin activity in living animals. Alis inhibited both plasma and tissue renin activity, ameliorated tissue oxidative stress, and attenuated UAE in a diabetes model. The in vivo imaging technique with the FRET technique will provide a novel approach to study the pathophysiology of kidney diseases.
Abstract 486: Uric Acid, Indoxyl Sulfate And Methylguanidine Activate The Bulbospinal Neurons In The Rostral Ventrolateral Medulla Through Their Specific Transporters
Authors: Oshima Naoki , Onimaru Hiroshi , Uchida Takahiro , Watanabe Atsushi , Takechi Hanako , Nishida Yasuhiro , Kumagai Hiroo
Published: March 2022

Chronic renal failure (CRF) is often accompanied with hypertension. However, the mechanism of hypertension in patients with CRF is not well known except the excess of body fluid. We considered that the mechanisms of hypertension in patients with CRF were due to increased activity of bulbospinal rostral ventrolateral medulla (RVLM) neurons caused by the stimulation of uremic toxins. The harmful effects of uremic toxins on the neurons of the bulbospinal RVLM have been scarcely examined until date. The aim of this study was to examine the effects of uremic toxins on the bulbospinal RVLM neurons electrophysiologically and histologically. To directly investigate whether RVLM neurons show sensitivity to uric acid (UA), indoxyl sulfate (IS), and methylguanidine (MG), we examined changes in the membrane potentials (MPs) of the bulbospinal RVLM neurons using the whole-cell patch-clamp technique, during superfusion of these toxins. During UA superfusion (0.05-1 mM), 29 of 33 RVLM neurons were depolarized, and during IS superfusion (0.01-5 mM), 14 of 14 RVLM neurons were depolarized. During MG superfusion (0.01-5 mM), 18 of 19 RVLM neurons were depolarized. To examine the transporters on RVLM neurons for these toxins, histological examinations were performed. As the results, all of 4 UA-depolarized RVLM neurons showed the presence of URAT 1. All of 3 IS-depolarized and four of 4 IS-depolarized RVLM neurons showed the presence of organic anion transporter (OAT) 1 and OAT 3, respectively. All of 3 MG-depolarized RVLM neurons showed the presence of organic cation transporter (OCT) 3. Our findings suggest that uremic toxins such as UA, IS, and MG increase the blood pressure by increasing the activity of the bulbospinal RVLM neurons through their specific transporters on these neurons.
Abstract 503: Maintenance of Normokalemia with ZS-9 Once Daily in CHF Patients on RAAS Inhibitors: Subgroup Analysis of a Phase 3 Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial of Patients With Hyperkalemia
Authors: El-Shahawy Mohamed A , Singh Bhupinder , Rasmussen Henrik S , Lavin Philip T , Yang Alex , Qunibi Wajeh
Published: March 2022

Risk of hyperkalemia (HK) limits the use of RAAS inhibitors (RAASi) in pts with congestive heart failure (CHF), chronic kidney disease (CKD), or diabetes. ZS-9, a nonabsorbed cation exchanger designed to preferentially entrap K
+
in the GI tract, significantly reduced serum K
+
vs placebo (PBO) over 48 hours with low rates of adverse events in CKD patients with hyperkalemia. A pre-specified subgroup analysis was conducted with CHF patients from a large Phase 3 trial of ZS-9 for HK.

Pts (N=753) with K
+
5.0-6.5 mmol/L were randomized (1:1:1:1:1) to ZS-9 (1.25, 2.5, 5 or 10 g) or PBO orally 3х daily for 48 hr (acute phase). At the end of this phase, pts with K
+
3.5-5.0 mmol/L (n=542) were randomized 1:1 to the same ZS-9 acute phase dose or PBO once daily (QD) on Days 3-15 (extended phase); patients initially on PBO were randomized to 1.25 or 2.5g ZS-9 QD. RAASi were kept constant during the study. Serum K
+
in CHF pts treated with the highest ZS-9 dose (10g) was compared to PBO with an unpaired t-test.

Among 753 pts, 300 (40%) had CHF at baseline (per investigator assessment), of whom 72% were on RAASi. Mean baseline K
+
was 5.3 mmol/L in the CHF pts. CHF pts who entered the Extended Phase (n=204) either remained on ZS-9 10g (n=26) or switched to PBO (n=23). The extended phase baseline K
+
was similar between groups (4.4 vs 4.5 mmol/L) and to the overall group (4.5 mmol/L ZS-9 [n=63] vs 4.4 mmol/L PBO [n=61]). On Day 15, mean K
+
for CHF patients on ZS-9 10g was 4.5 mmol/L vs 5.0 mmol/L for PBO (P=0.002; Fig).

ZS-9 10g once daily maintained mean serum K
+
<5.0 mmol/L in pts with CHF despite RAASi continuation in most pts, indicating that ZS-9 permits optimal cardiorenal protection with RAASi for this population.
Abstract 520: Na/H Exchanger Participates in Tubuloglomerular Feedback
Authors: Wang Hong , D’Ambrosio Martin A , Ren YiLin , Monu Sumit , Leung Pablo , Kutskill Kristopher , Garvin Jeffrey L , Peterson Edward L , Carretero Oscar A
Published: March 2022

The afferent arteriole (Af-Art) accounts for most renal vasculature resistance, thus controlling glomerular filtration and renal function. The nephron regulates Af-Art resistance via two mechanisms, the vasoconstrictor tubuloglomerular feedback (TGF), initiated in the macula densa via Na/K/2Cl cotransporters (NKCC2), and the vasodilator connecting tubule glomerular feedback (CTGF), initiated in the connecting tubule via the epithelial Na channels (ENaC). However, when TGF and CTGF are inhibited by furosemide and benzamil, a novel form of TGF is observed.

We hypothesize that in addition to NKCC2, TGF can be initiated by Na/H exchangers (NHE). Furthermore, we hypothesize that when NKCC2 and NHE are blocked, CTGF causes Af-Art dilation.

In vivo, using the nephron micropuncture technique, we performed two consecutive stop-flow pressure (PSF) curves (an index of glomerular capillary pressure) by increasing the perfusion of the nephron from 0 to 40 nL/min while adding drugs to the tubular perfusate.

TGF was blocked by furosemide (vehicle -7.9±0.2 mmHg at 40 nL/min, furosemide -0.4±0.2; P<0.001). An NKCC2-independent form of TGF was revealed by blocking TGF with furosemide and CTGF with the ENaC inhibitor benzamil (furosemide: -0.2±0.1 mmHg, furosemide+benzamil: -4.3±0.3 mmHg, P<0.001). TGF was completely blocked by the combination of furosemide and the NHE inhibitor dimethylamiloride (DMA), resulting in vasodilatation (furosemide+DMA: 2.7±0.5 mmHg, n=6), that was blocked by CTGF inhibition (furosemide+DMA+benzamil: -1.1±0.2 mmHg, P<0.01, n=6).

We conclude that NHE in the nephron causes TGF when NKCC2 and ENaC are inhibited and that CTGF causes dilation of the Af-Art when TGF is completely blocked with NKCC2 and NHE inhibitors.
Abstract 533: Effect of Dipeptidyl Peptidase-4 Inhibition on Blood Pressure and Its Dipping Pattern in Dahl Salt-Sensitive Rats
Authors: NISHIYAMA AKIRA , Fujisawa Yoshihide , Rahman Asadur , Nakano Disuke , Rafiq Kazi , Kobori Horoyuki , Sufiun Abu
Published: March 2022

Several studies have indicated that treatment with dipeptidyl peptidase-4 (DPP-4) inhibitors decreases blood pressure. We aimed to examine the effects of vildagliptin, a specific DPP-4 inhibitor, on blood pressure and its dipping pattern in Dahl salt-sensitive (DSS) rats. Male DSS rats were treated with a high salt (8% NaCl) diet and vehicle (0.5% carboxymethylcellulose) or vildagliptin (3 or 10 mg/kg, twice daily, p.o.) for 7 days (n = 7 per group). Mean arterial pressure (MAP) was measured by telemetry. High salt diet for 7 days significantly increased MAP with an extreme dipping pattern of blood pressure in DSS rats. Vildagliptin dose-dependently inhibited DPP-4 enzymatic activity and increased plasma GLP-1 levels. Furthermore, vildagliptin dose-dependently attenuated the development of salt-induced hypertension. Interestingly, vildagliptin significantly increased urine sodium excretion and normalized the dipping pattern. In anesthetized high salt-diet DSS rats, acute intra-cerebroventricular infusion of vildagliptin (50, 500 or 2500 μg in 10 μl solution) did not alter MAP or heart rate (n = 4). These data suggest that treatment with the DPP-4 inhibitor, vildagliptin, attenuates the extreme dipping pattern of blood pressure and development of salt sensitive hypertension by increasing urinary sodium excretion. These effects of vildagliptin may not be mediated through the central nervous system.
Abstract 540: Mineralocorticoid Receptor Antagonism Prevents The Aortic Dilatation And Extracellular Matrix Degradation Induced By Beta-adrenoceptor Overactivation
Authors: Clerici Stefano P , Victorio Jamaira A , Santos-Silva Junia C , Bessa Kleiber L , Ferrucci Danilo L , Carvalho Hernandes F , Rossoni Luciana V , Davel Ana P
Published: March 2022

Chronic stimulation of β-adrenoceptor (β-AR) and activation of renin-angiotensin-aldosterone system are common feature of cardiovascular diseases as heart failure and hypertension. β-AR overactivation induces cardiac remodeling, which contribute to cardiac dysfunction. However, it is unknown whether it alters arterial structure. Here, we demonstrated that β-AR overactivation induced by isoproterenol (ISO) treatment in Wistar rats (0.3 mg/kg/day, for 7 days) compared to control group (vs. CT, *p<0.05) increased thoracic aortic lumen area (CT: 1.8±0.06 vs. ISO: 2.3±0.08* mm2) and internal diameter (ID) (CT: 1.63±0.28 vs. ISO: 1.77±0.08* mm), without changes on wall thickness (wt). ISO caused disorganization and rupture of elastin fibers (Score; CT: 1.53±0.07 vs. ISO: 2.52±0.27*) and reduced aortic collagen deposition measured by picrosiuris red (CT: 56±3 vs. ISO: 32±2* A.U./vessel area). These alterations induced by ISO were associated with increased phosphorylation of extracellular signal-regulated kinase 1/2 (p-ERK/ ERK; CT: 0.75±0.07 vs. ISO: 0.95±0.05* A.U.); increased matrix metalloproteinase (MMP)-2 activity evaluated by zymography (CT: 4.0±0.9 vs. ISO: 10.8±1.2* A.U.); and reduced protein expression of TIMP-2 (CT: 0.95 ±0.07 vs. ISO: 0.48±0.08* A.U.) and collagen I/III (CT: 0.9±0.08 vs. ISO: 0.6±0.06* A.U.). ISO treatment did not change mean arterial pressure (MAP); however, it increased the mechanical forces circumferential wall tension (CWT= MAPxID/2; CT: 10.1±0.35 vs. ISO: 11.6±0.35* 103dyn/cm) and tensile stress (TS=CWT/wt; CT: 102±6 vs. ISO: 121±6* 104dyn/cm2) on aorta. The co-treatment with spironolactone (SPI, 200 mg/kg/day), a mineralocorticoid receptor (MR) antagonist prevented the ISO-induced aortic structural changes, MMP-2 and ERK1/2 activation, as well as the reductions on collagen and TIMP-2 expression. However, SPI did not modify MAP, CWS or TS. The results suggest that β-AR overactivation induces an aneurysm-like thoracic aortic remodeling characterized by vessel dilatation and extracellular matrix degradation, associated with MMP-2 and ERK1/2 activation. This remodeling and signaling were prevented by MR antagonist, independently of changes on aortic wall tension and tensile stress.
Abstract 547: Toll-Like Receptor 4 Signaling induces Systolic Blood Pressure Increase with Age
Authors: Baumann Marcus , Kemmner Stephan , Chmielewski Stefan , Aoqui Cristiane , Heemann Uwe
Published: March 2022

Background:
Systolic blood pressure (SBP) increases steadily with age in industrialized Western societies and bears an independent continuous relationship with the incidence of cardiovascular events. The molecular mechanisms linking age and SBP remain uncertain, but vascular aging is one suspected pathomechanism.

Vascular aging is characterized by increased aortic stiffness and promoted by oxidative stress. The origin of oxidative stress is multifactorial, but Toll-like receptor 4 (TLR4) as regulator of innate immunity is an important inducer. We hypothesized that TLR4 mediates oxidative burden during physiological aging, promotes vascular stiffness and increased systolic blood pressure.

Methods:
We investigated adult (6-9 months) and aged (12-15 months) C57Bl/6j and TLR4 null-mice (n=12/group). We determined blood pressure using tail-cuff plethysmography. Oxidative burden was determined by MDA. Vascular stiffness was assessed by aortic pulse wave velocity using the Complior and determination of aortic distensibility and compliance using ultrasound technique.

Results:
Here we show that the SBP increase with age (6±2mmHg) is abolished in aged TLR4 null-mice as compared to C57Bl/6j mice. With respect to vascular aging we show that despite similar aPWV in adult mice (2.2±0.2m/s) in aged TLR4 null-mice aortic stiffness is ameliorated (2.5±0.7m/s) as compared to C57Bl/6j mice (4.2±0.7m/s) and oxidative damage as determined by MDA was abolished in TLR4 null mice. Aortic distensibility reduced in both strains during aging. Aortic compliance was reduced with age only in C57Bl/6j mice, not TLR4 null mice.

Conclusion:
We suggest that the lack of TLR4 signaling reduced the aging associated cumulative oxidative burden. This ameliorated the development of aortic stiffness with age. The ameliorated vascular aging in TLR4 null-mice protected against the SBP increase with age.
Abstract 563: Whole Mitochondrial Genome Association Analysis Of Essential Hypertension In Chinese General Population
Authors: Li Yang , Chen Xi , Yang Jie , Gao Jinliao , Li Zongbin , Liu Yuqi , Zhang Yuxiao , Yin Tong
Published: March 2022

Objective:
Genetic studies of essential hypertension have mainly focused on the genome-wide association study of nuclear genes, which could explain just a small fraction of the heritability of blood pressure traits. This study aimed to investigate the relationship between mitochondrial genetic variants and essential hypertension (EH) in Chinese general population.

Methods:
Demographic and clinical matched 100 pairs of Chinese-Han EH patients and controls were recruited in the derivation cohort. A total of 797 EH patients and 816 controls were recruited in the derivation cohort. DNA was extracted from the peripheral whole blood of each patient. The whole mitochondrial genome was re-sequenced in the derivation cohort. The screened related variants were genotyped in the derivation cohort. Association analysis was performed to identify EH related variants.

Results:
In the derivation cohort, a total of 537 variants were detected in mitochondrial genome with 29 in tRNA-coding genes, 45 in rRNA-coding genes, 10 in D-Loop functional regions, and 453 in protein-encoded genes. No statistical difference could be found between EH patients and controls for the distribution of t-RNA, r-RNA and D-loop variants. For the protein-encoded genes, compared with controls, the occurrence of ND2 C5178A genetic variant was significantly lower [18% (18 in 100) vs. 31% (31 in 100),
P
= 0.03] in EH cases. Similarly, the distribution of C8414T of ATP8 gene in EH cases was significantly lower(10% vs. 22%,
P
= 0.02)than that in controls. No other mitochondrial variant exhibited significantly different distribution between EH cases and controls in the derivation cohort. The significant protective roles of ND2 and ATP8 genetic variants on EH were confirmed in the validation cohort (For ND2 C5178A: 24.59% (196 in 797) in EH vs. 32.72% (267 in 816) in control; adjusted OR: 0.62, 95% CI: 0.49-0.79,
P
< 0.001; For ATP8 C8414T: 17.69% (141 in 797) in EH vs. 23.16% (189 in 816) in control; adjusted OR: 0.68, 95% CI: 0.52-0.89,
P
= 0.005).

Conclusion:
This study provides the evidence that mitochondrial genetic variants of C5178A in ND2 gene and C8414T in ATP8 gene may exert protective role against EH in Chinese general population.
Abstract 574: The "Myotrophoblast": Endothelin-Induced Contraction in the Modified Spiral Artery of Rat Placenta
Authors: Ariel Ilana , Skarzinski Galina , Kossovsky Tally , Belzer Vitali , Knigin David , Kha'maisi Mogher , Abassi Zaid , Bursztyn Michael
Published: March 2022

We observed alpha-smooth muscle actin (αSMA) expression in rat endovascular trophoblasts (EVasT) and investigated the spatial and temporal expression of smooth muscle (SM) proteins and their potential function in remodeled spiral artery. Rat placentas were examined from gestational day 13 to term, and were immunostained for cytokeratin, αSMA, α heavy chain of SM myosin, myocardin (marker of SM differentiation) and endothelin receptors A and B (ETA, ETB: BQ-123 and BQ-788, respectively). Transverse sections of the modified spiral artery were studied
ex vivo
for endothelin-1-induced contraction. EVasT expressed SM proteins co-localizing with cytokeratin, confirming their trophoblastic origin, Thin fibers, consistent with actin fibers, were observed by transmission electron microscopy, in the cellular localization of αSMA in EVasT.Functional experiments revealed that addition of 10
-7
M endothelin-1
ex vivo
reduced vascular lumen cross section area by 10.1+/-0.9% compared with control. This effect was reduced to only 1.1+/-4.9% in the presence of ETA antagonist, and to 5.34.1% by ETB antagonist,
p
<0.001.

The expression of proteins involved in SM differentiation, regulation of function and contraction, along with the expression of the endothelin system, suggest that some vascular tone is potentially maintained by endothelin-1 in the rat remodeled spiral artery despite replacement of SM cells by trophoblasts. Endothelin-induced contraction of the modified spiral artery
ex vivo
is mediated via its receptors, suggesting its role in situations of dysregulation of the vasoactive systems.
Abstract 605: Brain ERK1/2 Mitogen-Activated Protein Kinase Mediated Phosphorylation of Potassium Channel Subunit Kv4.2 Likely Contributes to Sympathetic Activation by Blood-borne TNF-α
Authors: Yu Yang , Wei Shun-Guang , Zhang Zhi-Hua , Felder Robert B
Published: March 2022

Blood-borne tumor necrosis factor (TNF)-α acts upon the central nervous system to augment sympathetic drive in normal and pathophysiological conditions including heart failure. We have found that brain extracellular signal-regulated kinase (ERK)1/2 mediates TNF-α induced sympatho-excitatory responses, but the precise mechanisms are not understood. ERK1/2 can rapidly phosphorylate the voltage gated potassium channel (Kv) subunit Kv4.2 in neurons to reduce the transient outward potassium current that normally restrains neuronal excitability in cardiovascular-related brain regions. We hypothesized that ERK1/2 phosphorylation of Kv4.2 in the hypothalamic paraventricular nucleus (PVN) may contribute to TNF-α-induced sympathetic activity. In anesthetized rats, a centrally directed intracarotid artery (ICA) injection of TNF-α (0.5 μg/kg) increased (*p<0.05, vs baseline) renal sympathetic nerve activity, mean blood pressure and heart rate beginning as early as 30 minutes. These responses were reduced by a 4-hour continuous intracerebroventricular (ICV) infusion of the ERK1/2 inhibitor PD98059 (0.4 μg/hour). Molecular studies revealed that TNF-α-induced phosphorylation of ERK1/2 and Kv4.2 was detectable at 30 minutes (0.51±0.03* vs 0.28±0.03 for ERK1/2, 0.19±0.02* vs 0.00±0.00 for Kv4.2, TNF-α vs control, corrected by β-actin) and increased with time (4 hours after TNF-α: 0.65±0.01* vs 0.25±0.04 for ERK1/2; 0.30±0.02* vs 0.00±0.00 for Kv4.2). Interestingly, mRNA expression for nuclear factor (NF)-κB p65 or its inhibitor IκBα was unchanged at 30 minutes. However, 4 hours after TNF-α injection NF-κB p65 mRNA (1.77±0.11* vs 1.00±0.02 fold change) was augmented and IκBα mRNA (0.48±0.03* vs 1.00±0.07 fold change) was decreased. ICV PD98059 normalized phosphorylation of ERK1/2 (0.30±0.01*) and reduced phosphorylation of Kv4.2 (0.16±0.02*) as well as activation of NF-κB (1.34±0.11* for NF-κB p65; 0.70±0.08* for IκBα) in the PVN 4 hours after TNF-α injection. These data suggest that the acute sympatho-excitatory responses elicited by blood-borne TNF-α are mediated, at least in part, by brain ERK1/2-induced phosphorylation of Kv4.2, reducing the transient outward potassium current and increasing neuronal excitation.
Abstract 614: Anti-inflammatory Effect of Ang-(1-7) non-peptide mimetic (AVE 0991), on T Cell Infiltration in Perivascular Adipose Tissue in ApoE-/- Mice
Authors: Skiba Dominik , Nosalski Ryszard , Mikolajczyk Tomasz , Olszanecki Rafal , Jawien Jacek , Rios Francisco , Montezano Augusto C , Touyz Rhian M , Guzik Tomasz J
Published: March 2022

Angiotensin 1-7 (Ang 1-7) has anti-atherosclerotic effects, possibly through its anti-inflammatory properties. As perivascular inflammation is an important component of early atherosclerosis, we aimed to determine whether angiotensin-(1-7) non peptide mimetic (AVE0991) can modulate these inflammatory mechanisms in the early development of atherosclerosis. Twelve-week old ApoE-/- mice and controls (C57BL/6J) on chow diet were treated with placebo or AVE0991 (0.58μM/g/day, per os). Mice were analysed at 16, 20 and 24 weeks of age. The level of perivascular leukocyte and T cell infiltration was measured by flow cytometry. Atherosclerotic plaque area was assessed by Oil Red O staining of en-face aortic preparations. Endothelial dysfunction was measured by wire myography and ROS production in the aorta by lucigenin-enhanced chemiluminescence (5μM). RESULTS: Levels of endogenous Ang 1-7, measured by mass spectrometry in the perivascular adipose tissue were low in both WT and ApoE-/- (32.9±1.2 vs 31.1±1.1 pg/mg; p=0.1). AVE0991 significantly reduced plaque area in ApoE-/- mice at all time points (14.2±1.9% vs 7.25±1.3% at 24 weeks; p<0.05). During this early stage of atherosclerosis there was no evidence of endothelial dysfunction and no significant increase in ROS generation in ApoE-/- mice. However, there was significant perivascular inflammation in 16 week old ApoE mice: CD45+ leukocytes were 2 fold higher in ApoE-/- mice vs controls (3200±850 vs 1700±267 cells/mg; p<0.05) and the number of CD3+ T in ApoE-/- mice was higher than in C57Bl/6J (638±88 vs 310±64 cells/mg; p<0.05). With aging, the number of leukocytes and T cells further increased in ApoE-/- but not in C57BL/6J mice, an effect abrogated by AVE0991. Moreover in cell culture of SW872 preadipocytes, AVE 0991 preincubation abolished TNF-α induced increase of chemokine RANTES and IL-6 mRNA expression. CONCLUSION: Ang-(1-7) non-peptide mimetic (AVE 0991) effectively inhibits early perivascular inflammation during the development of atherosclerotic plaque. This occurs independently of changes in redox status or endothelial dysfunction.