bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P242: Microvascular Changes in Brain During Salt-sensitive Hypertension: Molecular Mechanisms and Implications for Small Vessel Disease
Authors: De Silva Travice M , Grobe Justin , Faraci Frank
Published: June 2021

Hypertension is a major risk factor for small vessel disease (SVD), a leading contributor to stroke and dementias. Mechanisms that underlie SVD in brain are poorly defined, with no specific therapy at present. Because parenchymal and pial arterioles are targets of the SVD process, we examined microvascular changes in a model using deoxycorticosterone-salt (DOCA) to activate the brain renin-angiotensin system (RAS), with resulting salt-sensitive (sodium- and fluid-dependent) hypertension. Male C57Bl/6 mice were treated with DOCA and given the choice of drinking H
2
O or H
2
O with 0.9% NaCl for 3 wks. Along with a modest elevation in mean arterial pressure (79±2 vs 95±3 mmHg, P<0.05), DOCA impaired endothelium-dependent dilation of both isolated parenchymal (baseline diameter of 15±1 μm) and pial arterioles (37±1 μm) in a pathway specific manner. Endothelium-dependent hyperpolarization was intact while eNOS-mediated vasodilation was markedly impaired along with reductions in phosphorylation in AKT (an upstream activator of eNOS). Local inhibition of angiotensin II type 1 (AT1-R) or mineralocorticoid receptors (MR) or Rho kinase (including ROCK2), restored endothelial function in DOCA-treated mice. Inner diameter of maximally dilated parenchymal arterioles was reduced approximately 20% by DOCA (P<0.05 vs sham). DOCA increased mRNA expression of RAS components (eg, AGT, ACE) in both brain and cerebral vessels. In NZ44 reporter mice that express GFP driven by the AT1A-R promoter, DOCA increased cerebrovascular GFP protein expression about 3-fold (P<0.05). Thus, DOCA activates both the brain and the cerebrovascular RAS, impairs select pathways affecting parenchymal and pial arteriolar function, while producing inward microvascular remodeling. AT1R, MR and ROCK2 are key contributors to cerebral microvascular dysfunction in this clinically relevant model of SVD.
Abstract P247: Non-dipping Ambulatory Blood Pressure in Non-hypertensive Individuals
Authors: Cheng Cynthia , Keith Scott
Published: June 2021

Non-dipping predicts increased cardiovascular risk in hypertension. However, the mechanisms leading to non-dipping remain unknown. Also, non-dipping in normotension and prehypertension is less well-described. The study objective was to determine the prevalence of non-dipping, and the association of non-dipping with other cardiovascular risk factors, in non-hypertensive young adults.

The study cohort included 176 non-hypertensive, non-diabetic individuals, mean age 27 (
Table 1
). Office and 24-hour ambulatory BP were measured, along with fasting adipokine levels, insulin sensitivity (QUICKI) and measures of vascular stiffness: Ambulatory arterial stiffness index (AASI) and pulse pressure. The distribution of dipping patterns was 58% dippers, 35% non-dippers, 5% extreme dipping, and 2% reverse dipping. CRP, IL6, and AASI are significantly associated with non-dipping (
Table 2
).

Non-dipping in common in young non-hypertensive adults. Inflammation and vascular stiffness may be potential mechanisms explaining the increased cardiovascular risk associated with non-dipping.
Abstract P248: Office and Ambulatory Brachial and Aortic Blood Pressure in Relation to Renal, Cardiac, and Vascular Organ Damage in Hypertension
Authors: Pareja Julia , Barrera Angela , Yun Sergi , Vazquez Susana , Oliveras Anna , Armario Pedro , Blanch Pere , Fernandez-Llama Patricia , Calero Francesca , De La Sierra Alejandro
Published: June 2021

It has been suggested that aortic BP could be a better predictor of target organ damage (TOD) than peripheral BP. However, studies examining such relationship by using ABPM are scarce. We aimed to examine such relationship in a cohort of hypertensive patients. A total of 208 patients (34% women; mean age: 57±12 years, 90% on antihypertensive treatment) were recruited from 4 hospital clinics in the area of Barcelona, Spain. The presence of TOD was determined if one of the following were present: cardiac, defined as the presence of LVH (LVMI ≥ 115 g/m
2
in men or ≥ 95 g/m
2
in women); renal, defined as the presence of either albuminuria (UAE ≥ 30mg/g) or reduced eGFR (< 60 ml/min/1.73 m
2
, by CKD-EPI formula), or both; vascular: aortic PWV ≥ 10 m/s. Clinic (mean of 4 measurements) and 24-h brachial and aortic BP were determined by the use of the portable automated Mobil-O-Graph device. LVH was present in 37%, albuminuria (17%), reduced eGFR (14%), or both were present in 26%, and elevated aortic PWV in 18%. Overall, 51% of patients presented TOD in at least one organ, whereas 6.3% have all renal, cardiac, and vascular alterations. After adjustments for age, gender, and the use of antihypertensive treatment, odds ratios for clinic, 24-hour, daytime, and nighttime brachial and aortic SBP and pulse pressure (PP) were all significantly associated with the presence of TOD. Higher odds ratios were observed for PP, compared to SBP, and for 24-hour estimates, compared to clinic, daytime or nighttime values. The OR for 5 mmHg increase in brachial and aortic PP were respectively 1.38 (1.16-1.60) and 1.41 (1.14-1.70). PP amplification or augmentation index (clinic or 24-h-derived) were not significantly associated with the presence of TOD, after adjustments. Considering specific organ alterations, aortic SBP and PP estimates were associated with LVH and aortic PWV, but not with albuminuria or reduced eGFR. Both aortic SBP and PP are associated with the presence of TOD in hypertensive patients. The association is stronger for 24-h PP and for LVH and arterial stiffness, whereas renal alterations show a weaker association. The use of ABPM including aortic BP measurement can be of interest in predicting cardiovascular alterations in hypertension.
Abstract P262: Spot Urine Sodium to Potassium Ratio Predicts Increasing Blood Pressure Levels in a General Population: The Nagahama Study
Authors: Tabara Yasuharu , Takahashi Yoshimitsu , Nakayama Takeo , Matsuda Fumihiko
Published: June 2021

Excessive salt intake is a risk factor for hypertension. The most reliable method for estimating daily salt intake is measurement of 24-h urinary sodium excretion, while it is inconvenient. Sodium-to-potassium ratio (Na/K) of a urine sample is another index of salt loading. We previously reported that a simple measure of spot urine Na/K might be a representative of salt loading in a cross-sectional setting. Here, we conducted a longitudinal study aiming to clarify a prognostic significance of spot urine Na/K for increasing blood pressure (BP) levels. Study subjects consists of 9,769 general individuals. Among them, individuals whose baseline Na/K was available (n=9,328), who were normotensive at baseline (n=6,392), and who participated in the follow-up measurement (n=5,209) were included in this analysis (51.8±12.9 years old, male: 29.2%). Mean follow-up duration was 5.0±0.5 years. Mean Na/K at baseline was 3.1±1.7, and showed step-wise increase with BP levels (optimal: 3.0±1.6, normal: 3.3±1.8, high normal: 3.4±1.8, P<0.001). Other major factors that were significantly associated with Na/K was fasting time (r=-0.220, P<0.001), and CKD (CKD (n=694): 2.7±1.6, control: 3.2±1.7, P<0.001). Mean SBP was significantly increased during follow-up period (baseline: 116±12, follow-up: 119±15 mmHg), and 805 individuals (15.5%) were newly diagnosed as hypertension (HT). These individuals were significantly older (HT: 60.3±9.9, NT: 50.3±12.8 years), were frequently male (36.4%, 27.9%), and had higher SBP (127±9, 115±11 mmHg) at baseline (P<0.001). In contrast, baseline spot urine Na/K was slightly lower in individuals who developed HT (3.0±1.6, 3.1±1.8, P=0.013), while that measured at follow-up investigation was oppositely higher in hypertensives (3.1±1.8, 2.8±1.5, P<0.001). Multiple linear regression analysis adjusted for the covariates identified baseline Na/K (β=0.108, P<0.001) and changes in Na/K during follow-up period (β=0.222, P<0.001) as independent determinants for future SBP levels. Higher spot urine Na/K, as well as increases in the Na/K levels, was significant determinant for future BP levels. The apparently lower baseline Na/K levels in individuals who developed HT might be due to reverse causality.
Abstract P264: DNA Methylation and Differences in Blood Pressure Levels in Monozygotic Twins
Authors: Kidambi Srividya , Li Yingchuan , Liu Pengyuan , leittl Michelle L , Coly Gerard , Cowley Allen W , Kotchen Theodore A , Liu Yong , Rockwell Cheryl E , Klump Kelly L , Burt S A , Gernaat Eric P , Donohue Mary F , Rayamajhi Supratik , Watson Ralph E , Mattson David L , Liang Mingyu
Published: June 2021

Background:
The development of common forms of hypertension (HTN) involves both genetic and environmental factors. Methylation changes (one of the epigenetic modifications) of DNA may play a role in the regulation of BP and development of HTN, and may result from interaction of specific genes with the environment. The current study investigates the relationship between genome-wide changes in DNA methylation in T-lymphocytes and BP level differences among monozygotic twins, who have identical DNA sequences.

Methods:
In a preliminary study, we recruited 24 pairs of monozygotic twins (60% women) with a mean age of 44 ± 10 years. Zygosity was determined via self-report or participants’ responses to a standard zygosity questionnaire. BPs were measured in triplicate after 5 minutes of rest at one minute intervals and averaged. Anthropometrics measurements were obtained along with blood for isolation of T-lymphocytes. DNA from T-lymphocytes was used to perform reduced representation bisulfite sequencing (RRBS) to measure methylation levels at single-base resolution in these subjects. Average differences in systolic (SBP) and diastolic (DBP) BPs were used as continuous variables for these analyses.

Results:
Mean SBP was 124 ± 15 (range (r): 97-174) mm Hg and mean DBP was 78 ± 11 (r: 49-106) mm Hg. Average differences in SBP and DBP among the co-twins (members of a twin pair) were 9 ± 10 (r: 0-40) mm Hg and 8 ± 6 (r: 0-26) mm Hg respectively. Average BMI was 29 ± 8 kg/m
2
. We observed that an average of 3063 (r: 757-7250) CpG islands were differentially methylated (DMRs) between co-twins. Among these DMRs, 10 were associated with average SBP difference and 5 were associated with average DBP difference at a significance level of p < 0.001. DMRs showing the strongest association (unadjusted p < 10
-4
) between the co-twins were located in the transcriptional start site (TSS) of genes:
CDC26
(cell division cycle protein) and
NR2F6
(nuclear receptor) for average SBP difference and
LEPRE1
(propyl 3 hydroxylase protein) for average DBP difference.

Conclusions:
BP differences between monozygotic co-twins, which most likely result from environmental factors, appear to be associated with differences in DNA methylation in T lymphocytes.
Abstract P265: Blood Pressure Control is Better and Less Expensive in Chronic Kidney Disease When Associated Metabolic Acidosis is Treated with Fruits and Vegetables Rather Than Sodium Bicarbonate
Authors: Goraya Nimrit , Simoni Jan , Pruszynski Jessica , Xiang Pin , Wesson Donald
Published: June 2021

Background:
Both sodium bicarbonate (NaHCO
3
) and base-producing fruits and vegetables (F+V) improve metabolic acidosis in chronic kidney disease (CKD) and appear to provide similar levels of kidney protection. Because F+V themselves reduce blood pressure, we examined if treatment of metabolic acidosis in CKD with F+V was associated with improved blood pressure control, using fewer anti-hypertensive drugs, and thereby with lower cost of hypertension management.

Methods:
We randomized 108 subjects with CKD stage 3 eGFR (30-59 ml/min) and metabolic acidosis as follows: F+V (n=36) added to reduce dietary potential renal acid load (PRAL) 50%, oral NaHCO
3
(HCO
3
, n=36) to reduce PRAL 50%, or no alkali (Usual Care, n=36). All were treated toward systolic blood pressure (SBP) <130 mmHg with regimens including ACE inhibition and followed 5 years.

Results:
Entry SBP and initial doses of 5 formulary anti-hypertensive drugs most commonly used for blood pressure control in CKD were not different among the 3 groups. At 5 years, SBP was lower in F+V (125±5 mm Hg) than both HCO
3
and Usual Care (135±5 and 134±5 mm Hg, respectively, p<0.01 vs. F+V for each). Daily doses for the following drugs at year 5 were lower in F+V than HCO
3
and Usual Care: Enalapril (8.3±2.4 vs. 11.1±3.6 and 11.7±4.8, mg/day, respectively, p<0.01), Diltiazem (1.7±7.0 vs. 145.8±36.0 and 153.3±35.7, mg/day, p<0.01), Clonidine (0.14±0.20 vs. 0.65±0.15 and 0.63±0.16, mg/day, p<0.01), Atenolol (0 vs. 6.25±15.1 and 6.25±15.1 mg/day, p<0.02) but there was no difference among groups in the year 5 dose of hydrochlorthiazide (16.1±9.9 vs. 21.9±16.2 and 21.5±16.3 mg/day, p=0.27). Five-year drug cost of hypertension management was less in F+V ($79,760) than both HCO
3
($155,372) and Usual Care ($152,305).

Conclusions:
Treating metabolic acidosis in CKD patients with F+V but not NaHCO
3
was associated with lower SBP, use of fewer and lower doses of anti-hypertensive drugs, and lower group cost of hypertension management. The data support that clinicians consider these adjunctive benefits of F+V on hypertension management when recommending treatment strategies for metabolic acidosis in CKD.
Abstract P267: Sex Differences in Cyp450 Activity and the Development of Hypertension and Renal Injury in the Dahl S Rat
Authors: Wu Wenjie , Murphy Sydney
Published: June 2021

It is well documented that a sexual dimorphism exists in the regulation of blood pressure in both the human population as well as experimental animal models, however evidence of a sex difference is lacking in the Dahl S rat. Thus, we hypothesize that alterations in CYP450 expression and 20-HETE production contribute to the progression of renal injury in Dahl S rats. Consistent with what we have previously reported, no difference was noted in the blood pressure of male or female Dahl SSJr rat (213.8±12 vs 196.8±13 mmHg, ns) following 4 weeks of a high salt diet (8%NaCl). However, proteinuria (148±25 vs 355±22 mg/day, p<0.05) and renal injury (1.9±0.01 vs 2.5±0.2) were lower in female relative to male rats. In addition, GFR was significantly reduced in male vs female rats (392.4±89 vs 829.5±98 μl/min/g, p<0.05) following at high salt challenge. Renal cortical (11.3.8±16 vs 20.99±2.8 pmol/min/mg, p<0.05) and outer medullary (19.4±3 vs 6.9±1.8 pmol/min/mg, p<0.05) 20-HETE production was elevated in female versus male rats. Furthermore, renal vascular 20-HETE production was elevated in the renal vessels compared to males (0.53±0.23 vs 3.2±1.2 pmol/min/mg, p<0.05). Thus, alterations in the production of renal eicosanoids may contribute to the delay in renal injury in females relative to male Dahl SSJr rats. AHA 14SDG20160020
Abstract P270: Carotid Stenosis: A Frequently Overlooked Etiology of Secondary Hypertension
Authors: Pannell J S , Gupta Mihir , Wali Arvin R , Abraham Peter , Porras Kevin A , Cheung Vincent J , Alam Yasaman , Khalessi Alexander A
Published: June 2021

Introduction:
Hypertension is one of the most prevalent etiologies for stroke, myocardial infarction, heart failure, retinopathy, and renal failure. Both primary and secondary hypertension can frequently be controlled and end organ damage be prevented by medications alone. However, atherosclerotic diseases resulting in end organ ischemic demand can overwhelm even the most aggressive treatment strategies, which have been demonstrated in cases of renal artery stenosis. However, carotid stenosis is an often-overlooked etiology in secondary hypertension, which can be treated by open surgery or endovascular stenting.

Methods:
In this retrospective study, the medical records of 43 consecutive patients who underwent carotid stenting for symptomatic or severe internal carotid artery stenosis from November of 2013 to December of 2015 were reviewed. All patients either had a prior stroke, TIA, or stenosis of the internal carotid artery measuring greater than 70% by NASCET criteria. All of the patients in the study had a diagnosis of hypertension. The systolic and diastolic blood pressures as well as the anti-hypertensive medication regimen of all of the patients involved were recorded before the surgery, immediately following surgery, and 3 months after the surgery. The average blood pressure reduction was calculated immediately following surgery and at 3 months.

Results:
The average reduction in systolic blood pressures immediately following surgery and at 3 months were 13.86 mmHg and 5.89 mmHg, respectively. The average reduction in diastolic blood pressures immediately following surgery and at 3 months were 8.96 mmHg and 1.84 mmHg, respectively. Additionally, the number of anti-hypertensive medications and the dosages decreased in 11 patients, including two of which who no longer required anti-hypertensives after stenting.

Conclusion:
Although the treatment of carotid artery disease is typically reserved for patients with cerebrovascular compromise, carotid artery stenting and endarterectomy can be useful adjunctives in the treatment of malignant and uncontrolled hypertension.
Abstract P272: Involvement of Neuro-Inflammation in the Pathogenesis of MCT-Induced Pulmonary Hypertension
Authors: Sharma Ravindra K , Shenoy Vinayak , Kumar Ashok , Mandloi Avinash , Katovich Michael J , Raizada Mohan K
Published: June 2021

Background:
Pulmonary hypertension (PH) is a devastating disease characterized by increase in pulmonary pressure that eventually leads to right heart failure and death. PH is associated with heightened circulatory cytokines and infiltration of inflammatory cells within the diseased lungs. However, involvement of inflammation within the central nervous system (CNS) in PH pathophysiology has never been investigated. Emerging evidence suggest that activated microglial cells and neuro-inflammation within the CNS play an important role in the pathology of several CNS disorders, including resistant hypertension.

Objective:
These observations led us to propose the hypothesis that microglial activation and neuro-inflammation in the autonomic brain regions play regulatory role in PH. Minocycline (Mino), an anti-inflammatory antibiotic, which has been reported to inhibit microglial activation in the autonomic brain regions, was used to test this hypothesis.

Methods:
PH was induced in adult male rats by a single injection of monocrotaline (MCT; 50mg/kg sc). A subset of MCT-injected animals was infused intracerebroventricularly (ICV) with Mino (20mg/ml). After 4 weeks of treatment, animals were sacrificed for the measurement of physiological and pathological parameters.

Results:
ICV infusion of Mino significantly attenuated right ventricular systolic pressure (RVSP; Con: 30.1±5, MCT: 76.1±14, MCT+Mino: 50.1±11 mmHg) and right ventricular hypertrophy (RVH; Con: 0.26±0.02, MCT: 0.49±0.12, MCT+Mino: 0.38+0.1) induced by MCT. MCT administration resulted in ~2 fold increase in microglial cells, predominantly in the hypothalamic paraventricular nucleus (PVN), an effect significantly attenuated by ICV Mino (Con: 4.0±1.0, MCT: 8.6±1.1, MCT+Mino: 5.0±1.0). MCT injection increased pro-inflammatory cytokines [IL-1β (155%), TNF-α (165%) and IL-6 (113%)] and decreased IL-10 (46%) levels in the PVN. However, ICV Mino treatment restored these cytokines to control levels.

Conclusion:
Our observations demonstrate that microglial activation in the PVN is involved in PH pathophysiology. They, for the first time, suggest the involvement of neuro-inflammation and autonomic dysregulation in the development and establishment of PH.
Abstract P279: Human Angiotensin Receptor Gene Variations, Hypertension & Reno-Vascular Complications
Authors: Jain Sudhir , Sirianni Natalie , Puri Nitin , Khudhair Ahmed A , Kumar Ashok
Published: June 2021

Angiotensin receptor type 1 (AT1R), a G-protein coupled receptor mediates the effect of angiotensin-II and contributes to the pathophysiological consequences of reno-vascular system. AT1R-signaling promotes renal sodium retention, vascular remodeling, hypertension, and end organ damage. Genetic variations that increase AT1R can cause pathological outcomes associated with renin angiotensin system overactivity. However, genetically variable, transcriptional regulation of the human AT1R gene is poorly understood. In this regard, the human AT1R gene has a haplotype block of four SNPs: T/A at -810, T/G at -713, A/C at -214, and A/G at -153 in its promoter. Variants -810T, -713T, -214A, and -153A always occur together (named haplotype-I or Hap-I) and variants -810A, -713G, -214C, and -153G always occur together (haplotype-II or Hap-II). We have found that hap-I is associated with hypertension in Caucasians. Thus, we generated transgenic (TG) mice with hap-II and I of the hAT1R gene to study its transcriptional regulation
in vivo
. TG mice with hap-I have higher baseline expression of hAT1R (3.9 folds) in the kidney with increased blood pressure (Hap-I, 126±3 vs. Hap II, 115±4). Since, diet-induced obesity is accompanied by systemic inflammation and redox imbalance that, in turn, alter the cellular transcriptional milieu, we gave Western diet (WD) treatment to our TG mice. Preliminary studies show that transcription factors like USF1, GR and STAT3 binds strongly (2.1; 2.3; 1.7 folds respectively Vs Hap-II) to increase hAT1R expression (5.8 folds) and resulting blood pressure (136±2 vs. 120±3 in Hap II) in TG-mice with hap-I, as compared to hap-II. Complementary experiments show increased inflammatory and redox markers in renal tissues of Hap-I mice, when compared to Hap-II, after WD; including, IL6 (5.9 fold), NOX1 (5.2 fold), CRP (9.8 fold), and TNFα (6.3 fold). Also, histochemical analysis of kidneys show an elevated pathology in Hap-I TG mice. Thus, haplotype-dependent transcriptional regulation of the hAT1R gene causes increased hAT1R expression and blood pressure, in Hap-I TG mice. Importantly, WD exacerbates this differential gene- expression regulation, further increasing hAT1R and promoting a pro-oxidant/inflammatory milieu in mice with Hap-I.
Abstract P285: Diastolic Dysfunction After Estrogen Loss is Linked to Cardiac Chymase in WKY but Not SHR Rats
Authors: Wang Hao , da Silva Jacqueline , Gabriel-Costa Daniele , Ahmad Sarfaraz , Sun Xuming , Lin Marina , Sudo Roberto T , Varagic Jasmina , Ferrario Carlos , Zapata-Sudo Gisele , Groban Leanne
Published: June 2021

Left ventricular diastolic dysfunction (LVDD) develops in response to hypertension and estrogen (E2) loss and is consequent to heart failure in women. To understand the mechanisms underlying the development of LVDD as a result of the interaction between E2 loss and the cardiac RAS, we compared the relationships of LV tissue RAS components and E/e′ between adult SHR (n=13) and WKY (n=9) female rats after ovariectomy (OVX) or sham surgery (intact). In intact rats, E/e′ was higher in SHR vs. WKY rats (P<0.05 strain effect) and after OVX, the diastolic phenotype of WKY’s mimicked that of intact SHR counterparts (Figure).

While relationships between RAS enzymatic activities and E/e′ were not significant in SHRs with respect to estrogen status (data not shown), OVX-induced increases in E/e′ were significantly linked to increases in chymase gene expression and enzymatic activity in the WKY strain (Figure). These data indicate that 1) the altered diastolic function in SHR is relatively insensitive to loss of estrogen while the opposite is true in WKY rats, and 2) OVX-induced LVDD in WKY is directly related to increases in cardiac chymase activity. Further elucidation of the interplay between an activated cardiac chymase-mediated RAS metabolism and LVDD following estrogen loss in normotensive subjects is warranted.
Abstract P291: High Fat Diet Promotes Hypertension and Vascular Remodeling but Not Vascular Fibrosis or Altered Contractility in Dahl Salt Sensitive Rats
Authors: Fernandes Roxanne , Perez Bonilla Patricia A , Garver Hannah , Galligan James J , Fink Gregory D , Xu Hui
Published: June 2021

Obesity associated hypertension in rodent models is commonly associated with altered vascular reactivity to sympathetic neurotransmitters and inflammation-induced vascular remodeling/fibrosis. Dahl salt-sensitive (SS) rats exhibit elevated sympathetic activity and vascular remodeling. We hypothesized that diet-induced obesity in Dahl SS rats would promote hypertension, vascular dysfunction and remodeling/fibrosis. Male Dahl SS rats were placed on high fat diet (HFD, 60% kcal from fat with final concentrations of 0.33% NaCl and 1% K
+
, n=5) or normal-fat diet (NFD; 10% kcal from fat, 0.24% NaCl, 0.36% K
+
, n=5) for 24-26 weeks after weaning (3 weeks of age). Compared with NFD rats, HFD rats displayed severe hypertension (MAP, 165±4 mmHg vs 133±6 mmHg, P<0.05), higher body-weight (470±6g vs 433±7g, P<0.05), and hyperlipidemia (cholesterol, 211±22 mg/dl vs 138±23 mg/dl, P=0.05). HFD rats did not show significant changes in plasma levels of fasting glucose (85±5 mg/dl vs 75±5 mg/dl), insulin (2.6±0.8 ng/ml vs 2.2±1.1 ng/ml), leptin (0.77±0.18 ng/ml vs 0.44±0.06 ng/ml), or aldosterone (249±3 pg/ml vs 234±3 pg/ml) (all P>0.05). HFD did not affect pressurized mesenteric arterial (~300 μm inner diameter, 60 mmHg) reactivity to norepinephrine or ATP
in vitro
. Pressurized mesenteric arteries from HFD rats displayed thicker walls (Ca
2+
free buffer, 40±1 μm vs 36±1 μm, P<0.05), but showed slightly increased distensibility. Morphological studies did not reveal greater fibrosis in adventitia of mesenteric, intrarenal and coronary arteries from HFD rats. However, HFD induced inflammation in mesenteric perivascular adipose tissue, as shown by increased CD3 positive cell infiltration and histological evidence of fibrosis and angiogenesis. Our studies indicate that HFD in male Dahl SS rats promotes hypertension, perivascular adipose tissue inflammation and vascular remodeling, but not vascular fibrosis. Alteration of vascular contractility to sympathetic neurotransmitters, however, is not required for obesity associated hypertension in Dahl SS rats.
Abstract P304: A Sympathetic-cholinergic Pathway Performs a Connection Between Brain and Spleen, to Prime Immune System and Induce Arterial Hypertension
Authors: Carnevale Daniela , Perrotta Marialuisa , Pallante Fabio , Carnevale Lorenzo , Cifelli Giuseppe , Fardella Valentina , Iacobucci Roberta , Fardella Stefania , Lembo Giuseppe
Published: June 2021

It is now widely recognized that immune system has a crucial role in hypertension. Various studies have demonstrated that the activation of adaptive immunity, and in particular of T cells, is a crucial moment in the onset and maintaining of hypertension induced by various stimuli in mice. Our previous studies have shown that hypertensive stimuli couple the sympathetic nervous system to determine the activation of splenic immune system. However, how the brain-to-spleen connection is realized in hypertension remains unknown. In this study we demonstrate that mice subjected to various hypertensive stimuli (AngII, DOCA-salt) show an increase of sympathetic nervous activity recorded in vivo in the splenic nerve (Firing Frequency: AngII 131±17 vs Veh 30±10 spikes/10 min, p<0.001). We also show how the sympathetic pathway induced by pro-hypertensive stimuli has its origin in the brain, converging into the spleen through a cholinergic-sympathetic connection that is realized through the vagus-splenic nerve drive and mediated at the molecular level by cholinergic nicotinic receptors at the level of celiac ganglion. In fact, we show that in celiac vagotomized mice, i.e. mice subjected to a procedure inhibiting vagal efferents but not central afferents, the splenic nervous drive induced by AngII was absent (AngII+VagX 21±4 vs AngII+sham 148±29 spikes/10 min, p<0.001). The same result was shown in α7 cholinergic nicotinic receptor KO mice, a receptor typically expressed by neurons in the peripheral ganglia (α7nAChR KO AngII 43±8 vs WT AngII 141±27 spikes/10 min, p<0.01). Moreover, we found that this cholinergic-sympathetic pathway was necessary to allow the activation of T cell costimulation and egression upon hypertensive challenges. Our results highlight a cholinergic-sympathetic pathway played by vagus-splenic nerves and responsible for immune system activation in response to hypertensive stimuli. We believe our results are significant because they reveal a previously unknown sympathetic pathway in hypertension for the first time. The brain-to-spleen connection realized through a cholinergic-sympathetic nervous drive that resembles the cholinergic anti-inflammatory pathway identified by immunologists in endotoxemia.
Abstract P307: Effect of Bradykinin Receptor Antagonism on ACE Associated Angioedema
Authors: Ramirez Claudia E , Straka Brittany T , Byrd James B , Stone Elizabeth , Woodard-Grice Alencia , Hui Nian , Yu Chang , Banerji Aleena , Brown Nancy J
Published: June 2021

The B2 receptor antagonist icatibant is approved for treatment of attacks of hereditary angioedema. Icatibant has been reported to decrease time-to-resolution of ACE inhibitor-associated angioedema in one study of European patients. Patients with ACE inhibitor-associated angioedema (defined as swelling of lips, tongue, pharynx or face during ACE inhibitor use and no swelling in the absence of ACE inhibitor use) were randomized within six hours of presentation to SQ icatibant 30 mg or placebo at zero and six hours later. Patients assessed severity of swelling using a visual analog scale serially following study drug administration or until discharge. Thirty-one patients were randomized to and received treatment, with 13 receiving icatibant and 18 receiving placebo. One patient randomized to icatibant did not complete the visual analog scale and was excluded. Two-thirds of patients were African American and two-thirds were women. Time-to-resolution of symptoms was similar in placebo and icatibant treatment groups [p=0.19 for the primary symptom (Figure) and p>0.16 for individual symptoms of face, lip, tongue, or eyelid swelling]. Frequency of administration of H1 and H2 blockers, corticosteroids, and epinephrine was similar in the two treatment groups. Lip swelling was more severe in blacks (p=0.03), while tongue swelling was more severe in whites (0.02). White patients were significantly more likely to receive epinephrine (p=0.01). Time-to-resolution of symptoms was similar in blacks and whites. This study does not support the clinical efficacy of administration of a B
2
receptor blocker in black or white American patients with ACE inhibitor-associated angioedema.
Abstract P309: Safety, Tolerability and Pharmacokinetic Data of the Novel Orally Active Formulation of Angiotensin- (1-7), Hydroxypropyl-β-cyclodextrin/ Ang- (1-7), in Healthy Volunteers- A Randomized Double-blinded Controlled Pilot Study
Authors: Koenen Janaina , Oliveira Marilene , Hamamoto Daniele , Santos Sérgio H , Sinisterra Ruben , Sousa Frederico B , Oliveira Junior Antônio R , Foscolo Rodrigo , Santos Robson A
Published: June 2021

Angiotensin-(1-7) is an endogenous peptide of the renin-angiotensin system in humans. It has several properties, which make it of great interest for healthcare, such as in systemic/ pulmonary hypertension treatment, insulin resistance improvement, reduction of visceral obesity and cardiac remodeling and arrhythmias. In this randomized double-blinded controlled phase I study we aimed to determine safety, tolerability, and pharmacokinetic properties of the new drug, angiotensin-(1-7) included in Hydroxypropyl β-cyclodextrin [Ang(1-7)/HPβCD)] in healthy adult volunteers, who were recruited in accordance to inclusion and exclusion criteria. Thirty-two volunteers ( 18-40 years-old ) were admitted to a hospital clinical research ward and remained under observation for two days, during which they received a single oral dose of Ang(1-7)HPβCD (equivalent to 0.35, 1.75 or 7.0 mg of the peptide) or placebo (N=8/ group). Vital signs and side effects were recorded according to the study protocol and blood samples were collected to obtain the pharmacokinetic profile of Ang-(1-7using LC/MS/MS. The oral administration of the novel compound Ang-(1-7)/HPβCD caused a dose-dependent elevation in the Ang-(1-7) plasma levels with a Tmax of 6.5±0.6 hours. The medians of the area under the curve for the placebo, 0.35, 1.75 and 7.0 mg doses were (in pg/mL/24 hours): 837.5±139.8 , 1094.4±224.3, 1415.0±187.7 and 1719.4±304.9, respectively. The Cmax for the 0.35, 1.75 and 7.0 mg doses were 26.1±3.17 pg/mL, 30.7±4.18pg/mL and 44.1±7.42 pg/mL, respectively. We did not find any statistically significant differences among the clinical and laboratorial parameters before and after the study. There were few mild side effects reported in the study, not related to the dose: two volunteers reported headache (one in the placebo group and one in the 0,35mg group) and three had dizziness and sweating in the orthostatic position (one in the 0.35 and two in the 1.75mg group). Our data show that the novel Ang-(1-7)/HPβCD formulation allow the absorption of Ang-(1-7), and is safe and well tolerated. Our results open new perspectives for future clinical trials with Ang-(1-7)/HPβCD for the treatment of arterial hypertension and other conditions.
Abstract P341: Osteoprotegerin-induced ROS Production and Redox Signalling is Exacerbated in Hypertension and Involves Syndecan-1 and TRPM2 Activation
Authors: Montezano Augusto C , Hepburn Ross , Graham Delyth , Touyz Rhian M
Published: June 2021

Osteoprotegerin (OPG) levels are increased in metabolic diseases, and are a biomarker of vascular dysfunction and cardiovascular risk. Mechanisms related to OPG-induced vascular dysfunction and its role in hypertension are not fully understood, but we previously demonstrated that OPG induces vascular dysfunction through ROS-dependent mechanisms. Here we assessed the molecular mechanisms whereby OPG regulates ROS and vascular function, with a focus on syndecan-1. VSMCs from normotensive (WKY) and hypertensive (SHRSP) rats were stimulated with OPG (50 ng/mL). ROS production was measured by lucigenin, amplex red and ELISA. In VSMCs from WKY rats, OPG increased ROS generation (158±15% vs veh, p<0.05). This effect was blocked by the syndecan-1 inhibitor (synstatin) and by removal of syndecan-1 sulfate proteoglycans side chains, chondroitinase and heparinase. OPG also increased H
2
O
2
(2 fold) and ONOO
-
(1.5 fold) levels in VSMCs (p<0.05). H
2
O
2
further stimulates ROS levels and redox signalling through activation of TRPM2, a redox-sensitive Ca
2+
channel. TRPM2 inhibitors, 8-bromo-ADPR (8Br) and N-(p-amylcinnamoyl)anthranilic acid (ACA), did not block OPG-induced ROS generation in VSMCs from WKY rats. Syndecan-1 activation leads to FAK and c-Src activation, which are redox-sensitive signalling proteins. FAK, but not c-Src, activation (117±2%, p<0.05) was observed after OPG stimulation of WKY VSMCs. In VSMCS from SHRSP rats, OPG effects on ROS generation were exacerbated (230±40%, p<0.05) and inhibited by synstatin, 8Br and ACA. OPG also increased FAK (118±2) and c-Src (113±1) activation (p<0.05) in VSMCs from SHRSP rats. In conclusion, OPG regulation of oxidative stress is increased in hypertension and involves not only syndecan-1, but also TRPM2 channels, which may lead to activation of redox-sensitive proteins and vascular damage.
Abstract P600: Activation Of GPER Ameliorates Pulmonary Hypertension In Female Rats
Authors: Zapata-Sudo Gisele , Alencar Allan K , Gabriel-Costa Daniele , Silva Ananssa M , Montes Guilherme C , Martinez Sabrina T , Fraga Aline M , Wang Hao , Groban Leanne , Sudo Roberto T
Published: June 2021

Introduction:
Pulmonary hypertension (PH) is primarily a disease of women (female-to-male ratio 4:1) and is associated with cardiac dysfunction.

Aim:
The activation of GPER by its agonist G1 was evaluated in monocrotaline (MCT)-induced PH rats.

Methods:
Depletion of estrogen was induced by bilateral oophorectomy (OVX; n = 18) in female Wistar rats (12 wks old). Experimental groups were: SHAM or OVX that received i.p. injection of MCT (60 mg/kg) for PH induction followed by administration of vehicle or G1 (400 μg/kg/day s.c.) for 14 days (n=7 per group). Hemodynamic parameters were determined by echocardiography. The effects of G1 in the maintenance of exercise capacity was investigated using a treadmill test.

Results:
MCT injection and estrogen loss led to a significant decrease in pulmonary acceleration time (PAT) and an increase in RV free wall thickness and MCT-related changes were attenuated by treatment with G1 (
P
< 0.05; Table 1). Right ventricular systolic pressure (RVSP) was higher in MCT-injected rats and the magnitude of this increase in OVX group was significantly higher than that in SHAM group. G1 normalized RVSP in both SHAM and OVX rats (Table 1). G1 treatment reversed altered expression of SERCA2a and phospholamban proteins in the RV (Table 1). Interaction between estrogen loss and MCT also reduced treadmill time to exhaustion in PH rats (
P
< 0.05), and chronic administration of G1 restored the exercise capacity (
P
< 0.05).

Conclusion:
G1 reversed PH-related cardiopulmonary dysfunction and exercise intolerance in female rats, a finding that may have important implications for the ongoing clinical evaluation of new drugs for the treatment of the disease in aging females.
Abstract P615: Stress-Induced Changes in Total Bone Mineral Content and Total Bone Mineral Density due to Activation of the Renin-Angiotensin-Aldosterone System in Adolescent Females
Authors: Pasham Vishwajeeth , Stewart Deborah , Carbone Laura , Harshfield Gregory A
Published: June 2021

Background:
Previous literature has shown a strong negative effect of angiotensin II (ANGII) on bone metabolism within mouse models. Additionally, psychological stress has been associated with activation of the renin-angiotensin-aldosterone system (RAAS). Stress has also been related to lower total bone mineral density (TBMD). However, there is controversy in the literature examining the relationship between the RAAS and bone metabolism within humans and stress has not been considered as a direct link between these systems.

Purpose:
We aimed to examine the relationship between stress-induced RAAS activation and TBMD and total bone mineral content (TBMC).

Methods:
Participants were placed on a sodium controlled diet for three days. Participants then underwent two hours rest, one hour mental stressor, and two hours recovery with hourly collections of blood/urine samples. Renin, ANGII, aldosterone, TBMD and TBMC were measured.

Results:
This study recruited 586 adolescents (mean age 16±1.116) with 51% women and 62% African-American and 38% Caucasian. Overall, relationships were observed between ANGII and aldosterone, and TBMC and TBMD controlling for age, race, and BMI. During stress, aldosterone was related to TBMD (r=-.150, p<0.05) and ANGII was related to TBMC (r=-.156, p<0.05) and TBMD (r=-.139, p<0.05). When comparing males and females, only females demonstrated a relationship between TBMC and ANGII in response to stress (stress: r=-.229, p<0.05; post-stress: r=-.277, p<0.01) and between aldosterone and TBMC (stress: r=-.199, p<0.05) and TBMD (stress: r=-.250, p<0.01). Renin was not significantly correlated with TBMD nor TBMC in any population.

Conclusion/Interpretations:
These data suggest that stress-induced RAAS activation may be associated with lower TBMD and TBMC in girls. Despite small correlations, consistency across multiple measures of RAAS activation being apparent in adolescents is significant. This observation may indicate that stress activation of RAAS contributes to bone remodeling in early life.
Abstract P618: Inactivation of Mitochondrial Deacetylase Sirt3 Promotes Vascular Oxidative Stress, Increases Endothelial Dysfunction and Exacerbates Hypertension
Authors: Dikalova Anna E , Uzhachenko Roman , Itani Hana A , Harrison David G , Dikalov Sergey
Published: June 2021

Endothelial dysfunction is associated with aging, diabetes, hyperlipidemia, obesity and these risk factors affect the expression and activity of the mitochondrial deacetylase Sirt3. Sirt3 activates major antioxidant SOD2 by deacetylation of specific lysine residues and Sirt3 depletion increases oxidative stress. We hypothesized that loss of vascular Sirt3 increases endothelial dysfunction, promotes hypertension and end organ damage. The role of vascular Sirt3 was studied in wild-type C57Bl/6J mice and tamoxifen-inducible smooth muscle specific Sirt3 knockout mice (Smc
Sirt3 KO
) using angiotensin II model of hypertension (Ang II, 0.7 mg/kg/day). Western blot showed 30% reduction of vascular Sirt3 and 2-fold increase in SOD2 acetylation in Ang II-infused WT mice. We have tested if ex vivo treatment of aorta with Sirt3 activator resveratrol improves endothelial function. Indeed, ex vivo incubation with resveratrol (10 μM) significantly reduced SOD2 acetylation, diminished mitochondrial O
2
and increased endothelial NO to normal level while Sirt3-inactive analog dihydroresveratrol had no effect. Specific role of vascular Sirt3 was studied in Smc
Sirt3 KO
mice by crossing floxed Sirt3 mice with mice carrying gene for inducible cre in the vascular smooth muscle. Sirt3 deletion exacerbates hypertension (165 mm Hg vs 155 mm Hg in wild-type) and significantly increases mortality in Ang II-infused Smc
Sirt3 KO
mice (60% vs 10% in wild-type) associated with severe edema and aortic aneurysm (100% vs 20% in wild-type). Decrease of NO is a hallmark of endothelial dysfunction in hypertension due to vascular oxidative stress. Indeed, Ang II infusion increased vascular O
2
by 2-fold and reduced endothelial NO by 2-fold. Interestingly, Ang II infusion in Smc
Sirt3 KO
mice caused severe vascular oxidative stress (3-fold increase in O
2
) and exacerbated endothelial dysfunction (4-fold decrease in NO). These data indicate that reduced vascular Sirt3 activity occurs in hypertension and this promotes vascular oxidative stress, increases endothelial dysfunction, exacerbates hypertension, increases end-organ-damage and mortality. It is conceivable that Sirt3 agonists and SOD2 mimetics may have therapeutic potential in cardiovascular disease.
Abstract P620: Aging Decreases Vascular GPER Expression and Function
Authors: Lindsey Sarah , Duong Jen L , Zimmerman Margaret A
Published: June 2021

Menopause accelerates the development of hypertension, arterial stiffness, end organ damage, and diastolic dysfunction. Postmenopausal hormone therapy relieves menopausal symptoms but promotes adverse cardiovascular outcomes, which may be due to aging-induced alterations in estrogen receptors. We previously published that GPER expression as well as agonist-induced vasorelaxation is decreased in mesenteric arteries from 12 month-old mRen2 female rats. We hypothesized that aging-induced GPER downregulation is present in other rodent species and strains as well. Aortas from young (2-3 months) and aged (11-23 months) C57BL/6 and BALB/c mice were lysed and immunoblotted for GPER protein. In addition, mesenteric arteries from C57BL/6 mice were mounted on a wire myograph and assessed for vasorelaxation in response to estrogen receptor agonists. We found a significant downregulation of GPER in aging aortas (3.1 ± 0.6 vs. 0.63 ± 0.42, P=0.016, N=4 per group). Moreover, we found that vasodilation to the GPER agonist G-1 was significantly attenuated in aging resistance arteries (22 ± 7.5% vs 58 ± 6.5%, P=0.023, N=2-4). Interestingly, vasodilation to the nonselective receptor agonist estradiol was not altered by aging (24 ± 10% vs. 32 ± 2.0%, P=0.23, N=2-4). In light of our previous findings in the rat vasculature, our data indicate that aging-induced decreases in vascular GPER expression and function are conserved across vascular beds and rodent species. We propose that aging-induced GPER downregulation switches the vascular benefits of postmenopausal estrogen therapy from positive to negative. Our future goal is to determine whether therapies that target GPER improve cardiovascular outcomes to protect the aging female population.
Abstract P628: The Influence Of Religiosity On The Embracement Vs. Technology Based Distance Learning In Therapy Adherence In Patient Hypertensive
Authors: Guerra Grazia M , Wen Chao L , Vieira Margarida , Fistarol Isabela , Tsunemi Miriam H , Giorgi Dante M , Motta Raquel A , Oliveira Jefferson C , Hong Valéria , Lopes Heno F , Consolim-Colombo Fernanda M , Bortolotto Luiz A
Published: June 2021

Introduction:
The approach known as ‘
embracement
’ adopts relational strategies or soft technologies which promote bonding and may impact therapy adherence

Objectives:
To assess the influence of the religiosity in the embracement approach on therapy adherence, quality of life, in hypertensive outpatients. This approach may be associated or not with the use of educational technology in a virtual learning environment(VLE) for distance learning (DL).

Methods:
This was a prospective randomized clinical study conducted with the following 3 groups of hypertensive patients: Group A (n=16, 12 women, mean age of 55.3±13 years, mean BMI of 32.3±6 kg/m
2
, receiving individual orientation required by an embracement strategy characterized by 7 nursing visits at 20-day intervals, for 4 months); Group VLE (n=10, 7 women, mean age of 51.5±7 years, mean BMI of 29.4±6 kg/m
2
,using a technological education strategy for DL and making 7 nursing visits at 20-day intervals, for 4 months); Control group (n=10, 5 women, mean age of 57.6±9 years, mean BMI of 29.7±6 kg/m
2
, making 1 nursing visit at baseline and 1 after 120 days.) At baseline and after 120 days, the following tools were applied: the Morisky test, WHOQOL, Religion Index (DUREL), and ambulatory blood pressure monitoring (ABPM). The VLE group had remote access to the ‘
Hypertension E-Care
’ site (6 specific educational modules).

Results:
At baseline, there were no differences in clinical blood pressure, ABPM, and socio-demographic variables among the 3 groups. At the final assessment, the VLE group (44.4±0.4) showed significant improvement (p<0.05) in the social domain of quality of life when compared to group A (40.8±4) and the controls (41.9±3);groups. In therapy adherence (Morisky test), the VLE group showed significant improvement at the end of the study, which was not the case with the other two groups. The significant correlations were observed between index of religiosity and the differences of BP Office for SBP R = - 0.667 , (p = 0.035 - negative correlation) and for DBP R =-0.666 (p = 0.035 - negative correlation) in VLE Group.

Conclusion:
This study shows that religious belief can improve blood pressure control, specifically when associated with education technology.