bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 483: Extracranial Artery Disease is Associated With An Impairment of Cardiac Autonomic Control
Authors: Pereira Valter L , Franqueiro Luiz F , de Godoy Moacir F , Campos Luciana A , Baltatu Ovidiu C
Published: March 2022

Background:
Atherosclerotic intima-media thickening (IMT) of carotid or subclavian artery may be associated with alterations in the sensitivity of carotid and subclavian baroreceptors. This is why we aimed at investigating if IMT is associated with alterations in the autonomic modulation of heart rate variability (HRV).

Methods:
This was a cross-sectional study with 30 subjects. The atherosclerotic artery IMT was determined by B-mode and duplex ultrasonography. HRV was evaluated through linear (low frequency [LF, ms2], high frequency [HF, ms2], standard deviation (SD) of normal R-R wave intervals [SDNN], square root of the mean of the sum of the squares of differences between adjacent normal R wave intervals [RMSSD]) and nonlinear Poincaré analysis (SD of the points perpendicular to the line-of-identity [SD1, ms]; SD along the line-of-identity [SD2, ms]).

Results:
A significant negative linear correlation was identified between the IMT of common carotid artery and the HRV indexes that reflects baroreflex and the fluctuations of autonomic tone: LF, Spearman's r = -0.52, 95% CI: -0.75 to -0.19, P = 0.003; SDNN, Spearman's r = -0.57, 95% CI: -0.78 to -0.25, P = 0.001; SD2, Spearman's r = -0.59, 95% CI: -0.79 to -0.28, P = 0.0006. Similar strong negative correlations were found for subclavian IMT. In contrast, there was no relationship between IMT and the HRV indexes related to autonomic tone (HF, LF/HF, RMSSD, SD1, SD1/SD2). The receiver operating characteristic (ROC) sensitivities/specificities/area under curve of SD2 for predicting carotid and subclavian artery IMT were 0.75/0.83/0.81 and 0.70/0.69/0.70, respectively (cutoff values: <32.8 and <42.5, respectively). Identification of atherosclerotic plaque in subclavian artery (IMT > 1.5 mm) was associated with a drastic reduction of SD2 (25.9 [24.3-40.5] vs. 50.2 [38.1-68.7]; data are median and interquartile range values, p = 0.0009), LF (65% lower, p = 0.005) and SDNN (43% lower, p = 0.001).

Conclusions:
Our data demonstrate that the atherosclerotic carotid and subclavian IMT are associated with an impairment of cardiac autonomic control. The SD2 index of Poincaré plot may be a functional biomarker for subjects at risk for carotid disease and an indicator for ultrasound evaluation.
Abstract 484: Augmented Myogenic Responses in Cerebral Arteries Following Interference with Peroxisome Proliferator-Activated Receptor γ (PPARγ) in Smooth Muscle via a Rho Kinase (ROCK)-Dependent Mechanism
Authors: De Silva Michael , Ketsawatsomkron Pimonrat , Pelham Christopher , Sigmund Curt D , Faraci Frank M
Published: March 2022

Myogenic responses by resistance vessels in brain play a crucial role in regulating cerebral blood flow and protecting the brain parenchyma and blood-brain barrier against potentially detrimental elevations in blood pressure. PPARγ is a transcription factor that may protect the vasculature by suppressing increases in vascular tone. Using transgenic mice expressing a dominant negative mutation of human PPARγ only in smooth muscle cells (S-P467L; TG), we have shown that augmented myogenic responses in mesenteric arteries are mediated by a mechanism that involved protein kinase C, but not ROCK. As ROCK is thought to play a major role in cerebrovascular disease, we hypothesized that interference with PPARγ in smooth muscle would augment myogenic responses in cerebral arteries via a ROCK dependent mechanism. We studied S-P467L TG mice and non-transgenic littermates (NT). Middle cerebral arteries (MCA) from TG mice generated more myogenic tone compared with NT (75 mmHg: 32±2% vs. 13±2% of max diameter). Myogenic reactivity was similarly augmented in MCA from TG mice from 15-150 mmHg whereas vessel diameter in Ca2+ free conditions was similar between groups. We used two approaches to evaluate the impact of ROCK in this model. First, the ROCK inhibitor, Y27632 (3 μM), abolished the augmented myogenic tone in MCA from TG mice (34±5% vs. 9±4% of max diameter). Second, treatment with a cell permeable molecule that modifies RhoA to make it constitutively active (CN-03) elevated myogenic tone to ~60% in both groups, which was reduced to ~10% by Y27632. The large conductance potassium (BK) channel modulates myogenic tone. The BK channel inhibitor, iberiotoxin, caused greater constriction in MCA from NT compared with TG (100 nM: NT, 14±2% vs. TG, 8±1% Δ tone at 75 mmHg). Although angiotensin II can activate ROCK, the AT1 receptor antagonist (losartan) did not affect myogenic tone in MCA from TG mice. Gene expression analysis revealed no change in mRNA for RhoA or ROCK1/2 expression in cerebral arteries from TG mice. These findings support the concept that in cerebral arteries, activity of ROCK has substantial effects on myogenic tone. The data also suggest that under normal conditions, PPARγ in smooth muscle inhibits ROCK, influencing myogenic tone in resistance arteries in brain.
Abstract 489: Hydrogen Peroxide Stimulates Renin Release From Mouse Juxtaglomerular Cells by Enhancing cAMP
Authors: Mendez Mariela
Published: March 2022

Renin plays an essential role in blood pressure control and the development of hypertension. Enhanced hydrogen peroxide (H
2
O
2
) levels in the kidney are associated with hypertension enhancing tubular salt transport and vascular reactivity. However, it is not known whether H
2
O
2
can directly stimulate renin release from renal juxtaglomerular (JG) cells. Since cAMP is one of the most potent signaling messengers for renin release, we hypothesized that H
2
O
2
stimulates renin release by enhancing cAMP production in JG cells. Primary cultures of mouse JG cells were treated for 60 min with different agonists and inhibitors and renin released to the supernatant measured by radioimmunoassay. We first tested the effect of decreasing endogenous H
2
O
2
with catalase. Catalase (100U/ml) blunted baseline renin release by 45±9% (
n
= 6;
p
< 0.05) indicating that endogenously produced H
2
O
2
tonically stimulates renin release. We then tested the effect of exogenous H
2
O
2
for 60 min on renin release. Physiological concentrations of H
2
O
2
(0.1 and 1μM) increased renin release by 101±43% (
p
= 0.08) and 166±47% (
p
< 0.04), respectively. To assure that exogenous H
2
O
2
crosses the plasma membrane to activate signaling, we monitored intracellular H
2
O
2
with a genetically encoded sensor (Hyper). We found that addition of exogenous H
2
O
2
enhanced intracellular H
2
O
2
within 5 minutes (
p
< 0.02; n=5). cAMP is the most potent second messenger for stimulation of renin release. Thus, we tested whether H
2
O
2
stimulates intracellular cAMP levels. For this, JG cells were transduced with a cAMP-sensitive FRET (CFP/YFP) probe (Epac2camps) and cAMP measured by confocal microscopy in real-time. Treatment of JG cells with H
2
O
2
increased cAMP within 30 min as indicated by a 22±3% increase in FRET (
p
< 0.03, n=3) whereas forskolin/IBMX (10μM/0.5mM) caused a maximal FRET stimulation of 44±13% (
p
< 0.01; n=5). Inhibition of adenylyl cyclase with SQ22536 (100 μM) completely blocked H
2
O
2
-induced increase in cAMP (N.S from baseline; n=6). We concluded that H
2
O
2
is a potent stimuli for renin release from mouse JG cells and this effect is most likely mediated by enhanced production of cAMP. Our data suggest that enhances renal cortical reactive oxygen species may induce hypertension by enhancing renin release.
Abstract 494: Loss of Gstm1 Alters Blood Pressure Homeostasis and Augments Angiotensin II-induced Hypertension
Authors: Cechova Sylvia , Chan Rosa , Koller Beverly , Le Thu H
Published: March 2022

GSTM1 gene encodes an enzyme that belongs to a superfamily of Glutathione-S-transferases that metabolize xenobiotic, and a broad range of reactive oxygen species and electrophilic compounds formed as secondary metabolites during the oxidative stress. There is a general consensus that oxidative stress contributes to development of hypertension (HTN). We hypothesized that
Gstm1
may influence blood pressure (BP) regulation through its central role in modulating redox homeostasis. Therefore
the objective of this study
was to determine the response of
Gstm1
knockout (KO) mice to 2 models of hypertension that are mediated in part by oxidative stress: 1) high salt model, and 2) angiotensin II model.

Methods:
Gstm1-/-
(KO) mice were generated on the 129S6 background through conventional gene targeting strategy. Hypertension was induced using high salt diet (HSD, 6% NaCl) for 2 weeks or by infusion of angiotensin II (Ang II) via mini-osmotic pump at dose 1000 ng/kg/min for 3 weeks. Systolic BP (SBP) at baseline and under high salt condition was measured using radio-telemetry. Tail-cuff manometry was used to measure SBP in Ang II induced hypertension. Urinary isoprostane (index of oxidative stress) was measured using ELISA assay. For all studies, n ≥ 4 per group.

Results:
By radiotelemetry,
Gstm1
KO mice display a modest but significantly higher baseline SBP compared to their wild type (WT) littermates: KO: 138.8 ± 1.3; WT: 132.1 ± 1.1, p < 0.01. On high salt diet, both WT and KO mice displayed salt sensitivity, and the difference in SBP was maintained between the two genotypes (KO: 148.7 ± 1.6; WT: 143.8 ± 1.3, p < 0.05). Urinary isoprostane (ng/100 mg of body weight) is significantly higher in KO mice than WT mice at baseline as well as during HSD (baseline: KO 15.1 ± 2.9; WT: 8.0 ± 0.8, p < 0.04; HSD: KO 27.2 ± 2.0; WT: 21.0 ± 1.3, p = 0.04). Angiotensin II infusion augments the difference in SBP between WT and KO mice (185.0 ± 3.1; WT mice 175.4 ± 1.4, p < 0.05).

Conclusion:
Loss of
Gstm1
results in elevated baseline blood pressure and augmented Ang II induced hypertension. The altered blood pressure homeostasis from deficiency in GSTM1 enzyme may be mediated by a decreased capacity to handle oxidative stress.
Abstract 504: Muscle Regeneration is Impaired in Diabetic Mice
Authors: Mogi Masaki , Kohara Katsuhiko , Nakaoka Hirotomo , Tsukuda Kana , Wang Xiaoli , Ohshima Kousei , Chisaka Toshiyuki , Bai Huiyu , Kan-no Harumi , Min Li-Juan , Iwanami Jun , Miki Tetsuro , Horiuchi Masatsugu
Published: March 2022

Background
Sarcopenia is the degenerative loss of skeletal muscle mass which impaired quality of life in the elderly. On the other hand, sarcopenic obesity which is a loss of muscle and a concomitant increase in fat is reported to increase in life-style related diseases. Especially the prevalence of sarcopenia is greater in patients with type-2 diabetes mellitus (T2DM) than in non-diabetic subjects. However, the detailed mechanism of such muscle degeneration has not been well established. Here, we investigated muscle regeneration using muscle-injured models in life-style related disease mouse model such as angiotensin II overexpression mice for hypertension and obese diabetic mice for diabetes.

Methods
Male eight-week-old wild-type mice (C57BL6), Tsukuba hypertensive mice (transgenic mice carrying human renin and angiotensinogen genes) and T2DM mice (KKAy) were undergone intramuscular injection of cardiotoxin (Ctx) (100uL/10uM) into tibia. After two weeks, muscle were removed and stained with hematoxylin and eosin. KKAy replaced by bone marrow prepared from GFP-transgenic mice were generated by 8 Gy whole body X-ray irradiation.

Results
Treatment with Ctx showed increased muscle satellite cells where nucleuses were observed in the center of cells. In Tsukuba hypertensive mice, inflammatory cells were significantly accumulated in muscle compared with C57BL6, but muscle regeneration was not significantly impaired. KKAy mice exhibited impaired muscle regeneration. Tibia weight was a half in Ctx-injected KKAy compared with Ctx non-injected KKAy, and Ctx-injected and non-injected C57BL6 tibia. Moreover, remarkable fat deposition was observed in Ctx-treated muscle of KKAy. Such change was also observed in another diabetic mouse model, db/db, but not in streptozocin-induced diabetic mice. Fat deposition was remarkably increased in aged KKAy (6-month-old) compared with younger mice (8-week-old). In GFP-chimeric mice, fat tissue showed highly GFP-positive indicating that fat tissue was generated from marrow cells.

Conclusion
Diabetic mice showed significantly impaired muscle regeneration with fat deposition, suggesting that diabetes enhanced sarcopenic obesity possibly due to inhibitory effect of marrow cell differentiation.
Abstract 506: Effects of High Salt Diet on Urine and Sodium Excretion in Db/db Mice
Authors: Obih Patience O , Ezebuenyi Michael , Jimoh Akeem , Summers Brittini
Published: March 2022

Cardiovascular diseases are among the complications of diabetes mellitus. People with hypertension are encouraged to reduce salt intake. Our working hypothesis is that diabetes will not cause sodium retention in the presence sodium load. This study was carried out to evaluate the role of diabetes in pressure diuresis and natriuresis. Diabetic mice (db/db-/-) 5 week old adult (37-49 g) male and diabetic were purchased from Jackson Laboratory, Bar Harbor, ME, USA. The db/db phenotype spontaneously develops characteristics of type 2 diabetes including obesity, hyperinsulinemia, and insulin resistance. The animals were randomly divided into groups (n=5-6 per group) and assigned to normal rat chow (0.5% NaCl diet) or high salt diet (8% NaCl from Research Diets, Inc. New Brunswick, NJ, USA), for 7 weeks. Renal function was determined by measuring urine volume (Uv), urinary and sodium (UNav). Metabolic cages were used in collecting 24-hr urine samples. Urinary sodium was measured using flame photometer. There were no significant differences in urine volumes collected on Day 1 in high salt and normal salt-fed db/db-/- mice. Urinary volumes (ml) of 26.7±8.1, 18.7±3.9, 28.0±3.5 of high salt-fed mice and 22.5±4.7, 18.2±5.9, 26.0±4.2 of normal salt-fed mice were collected in weeks 1, 3 and 7. The observed weekly increase in urine volume was significant in each group, but not significant between the two groups. Urinary sodium (μmol/ 24 hr) of 16.6±7.1, 22.8±4.1, 66.1±21.3 of high salt-fed mice and 1.0±0.2, 12.5±3.4, 3.5±0.1 of normal salt-fed mice were collected in weeks 1, 3, and 7. There were significant weekly increases in sodium excretion in high salt diet only. From our observation, sodium was excreted significantly. Therefore, sodium retention may not be a contributory factor to the incidence of hypertension in diabetes. There is need for further investigations on the complications of diabetes in relation to cardiovascular events such as hypertension.
Abstract 509: Role of TRPV1 and TRPV4 Channels in the Renal Vasculature
Authors: Chen Lan , Kaßmann Mario , Marko Lajos , Tepel Martin , Gollasch Maik
Published: March 2022

Transient receptor potential vanilloid 1 (TRPV1) and vanilloid 4 (TRPV4) cation channels have been recently identified to promote endothelium-dependent relaxation of small mesenteric arteries of mice, with calcium sparklets providing elementary calcium influx through single TRPV4 channels. The role of TRPV1 and TRPV4 in the renal vasculature is largely unknown. We hypothesized that TRPV1 and TRPV4 play an important role in endothelium-dependent vasodilation of renal blood vessels. We performed myography experiments on isolated renal arteries and perfused isolated kidneys of C57BL/6J mice. Isolated mesenteric arteries served as control vessels. The TRPV1 agonist capsaicin relaxed mesenteric arteries with an EC
50
of ~9 nM, but not large renal arteries (EC
50
>10 μM). This relaxation was inhibited by the TRPV1 antagonist capsazepine (10 μM). The capsaicin effect was absent in vessels of TRPV1 knock-out mice. Cytochrome P450 (CYP) 4A- and CYP4F-derived hydroxyeicosatetraenoic acid (20-HETE), which is a proinflammatory mediator of endotoxin-induced acute systemic inflammation, activated TRPV1 channels in patch clamp studies. The TRPV4 agonist GSK1016790A relaxed renal and mesenteric arteries with EC
50
of ~12 nM and 70 nM, respectively. These effects were inhibited by a specific TRPV4 antagonist, AB159908 (10 μM). Capsaicin

and GSK1016790A produced vascular dilation in isolated kidneys with EC
50
of ~11 nM and ~2 nM, respectively. The capsaicin effects were absent in kidneys of TRPV1 knock-out mice, whereas the effects of

GSK1016790A were also observed in TRPV1 knock-out mice. Our results demonstrate that TRPV4 channels are capable to promote endothelial relaxation in large and small renal arteries, whereas TRPV1 channels play a specific vasodilatory role in small arteries of the kidney. We suggest that specific activation of TRPV1 and TRPV4 may be a novel promising strategy for treatment of sepsis-associated renal vasoconstriction.
Abstract 51: Nebivolol, but not Metoprolol, Lowers Blood Pressure in a Human Model of Hypertension Sensitive to Nitric Oxide
Authors: Okamoto Luis E , Gamboa Alfredo , Shibao Cyndya , Arnold Amy C , Choi Leena , Black Bonnie K , Raj Satish R , Robertson David , Biaggioni Italo
Published: March 2022

Nebivolol, unlike other selective β1-receptor blockers, induces vasodilation attributable to increased nitric oxide (NO) function. Autonomic function normally masks the contribution of this effect because of baroreflex buffering. We have previously shown that sildenafil lowers blood pressure (BP) in autonomic failure patients with supine hypertension. These patients, therefore, provide a unique model of hypertension devoid of autonomic modulation but sensitive to increases in NO function. To test the hypothesis that nebivolol would decrease BP in this patient population through a mechanism independent of β-blockade, we randomized 20 autonomic failure patients with supine hypertension (14 men, 69±2 yr) to receive a single oral dose of placebo, nebivolol 5 mg, metoprolol 50 mg (negative control) and sildenafil 25 mg (positive control) on separate nights in a double-blind, crossover study. Medications were given at 8 pm. Supine BP was monitored every 2 hr for 12 hr. Compared to placebo, sildenafil and nebivolol decreased systolic BP (SBP) during the night (P<0.001 and P=0.036, by mixed-effects model) with a similar maximal reduction in SBP 8 hr post drug (-20±6 and -24±9 mm Hg, respectively. Figure). Metoprolol, in contrast, had no significant effect on BP. Despite lowering nighttime BP, nebivolol did not worsen orthostatic tolerance the following morning (8am) compared to placebo. We conclude that nebivolol effectively lowers BP by a mechanism that does not involve β1-blockade and to a similar magnitude to that produced by NO potentiation with sildenafil. Nebivolol might be a useful alternative to treat patients with this condition.
Abstract 510: Dietary Restriction Improves Vasodilator Dysfunction Caused by Accelerated Vascular Aging Due to Genomic Instability
Authors: Wu Haiyan , Durik Matej , Reiling Erwin , Danser Jan , Hoeijmakers Jan , Dollé Martijn , Roks Anton
Published: March 2022

Objective
Recently we discovered that genomic instability is associated with accelerated vascular aging in humans and mouse models, and that DNA repair-defective mice (

ERCC1
-/

Δ

7

) display accelerated age-dependent deterioration of endothelium-dependent and vascular smooth muscle dilator function (Durik M. et al. Circulation 2012).

Dietary restriction (
DR
) is known to slow aging. We explored whether
DR
would inhibit accelerated vascular aging caused by genomic instability by measuring vasodilator function in both male and female

ERCC1
-/

Δ

7

mice vs. wild-type (WT) littermates.

Methods
WT and ERCC1
-/
Δ
7
were fed with
DR
(increasing 10 to 30% nutrient restriction) or ad libitum (
AL
)
diets for 9 weeks, whereafter

thoracic aortas were isolated and used for
ex vivo
organ bath experiments. To investigate endothelial and vascular smooth muscle dilator function, aorta preconstricted with U44619 (thromboxane analogue) was exposed to acetylcholine (Ach: 10
-9
-10
-5
M) and sodium nitroprusside (SNP: 10
-9
-10
-4
M), respectively. Maximal dilator responses (mean+/-SEM) are shown between brackets, and were calculated as % decrease of precontraction. Significance values are those of dose-related responses tested by general linear model for repeated measures.

Results
In ERCC1
-/
Δ
7
mice fed
AL
both ACh and SNP responses were significantly smaller than in WT (Ach: 66.89+/-8.9% (n=8)
vs.
41.45+/-4.74% (n=11); SNP: 80.27+/-4.89% (n=5)
vs.
59.35+/-3.32% (n=5),
p
<0.0001, WT
vs.
ERCC1
-/
Δ
7
) with no gender differences. Whilst without effect in WT,
DR
normalized the SNP responses of ERCC1
-/
Δ
7
(79.60+/-7.09% (n=6)) to that of WT gender-independently. ACh responses were also improved (from 41.45+/-4.74%
AL
(n=11) to 56.02+/-4.73%
DR
(n=11)), but most pronounced in female ERCC1
-/
Δ
7
(38.33+/-8.39%
AL
(n=4)
vs.
-67.31+/-6.22%
DR
(n=4)
, p
<0.0001).

Conclusion
In ERCC1
-/Δ7
mice, DR improved vasodilator response in both genders. Endothelial function might be improved only efficiently in female mice, whilst the NO-mediated dilation of VSMC is improved equally well in both genders. Therefore,
DR
is beneficial for vascular function during vascular aging caused by genomic instability, and its impact on EC versus VSMC appears to be gender-dependent.
Abstract 514: Females are Protected from Hypertension and Kidney and Brain Infiltration of T Lymphocytes During Angiotensin II Infusion
Authors: Pollow Dennis , Uhrlaub Jennifer , Brooks Heddwen , Nikolich-Zugich Janko , Hay Meredith
Published: March 2022

There is extensive evidence that activation of the immune system is both necessary and required for the development of Ang II induced hypertension in males and that the central nervous system (CNS) is involved in this response. The purpose of the present study was to determine if there are sex differences in ability of the adaptive immune system to induce Ang II hypertension and the effect of sex on T-cell infiltration into the brain and kidney during Ang II induced hypertension. Rag-1-/- mice, with a genetic deletion of the recombinase-activating gene and lacking both T and B cells were used in this study. Male (n=4) and female (n=4) Rag-1-/- mice both received adoptive transfer of male CD3+ T cells three weeks prior to 14 day Ang II infusion (490 ng/kg/min). Blood pressure was monitored non-invasively via tail cuff. In Rag-1-/- mice, in the absence of T cells, male and female mean arterial pressure (MAP) responses to Ang II were similar, males increased MAP 14.6 mmHg (107.8mmHg to122.5mmHg) and females increased 14.4 mmHg (102.7mmHg to 117.2 mmHg). However, following adoptive transfer of male CD3+ Tcells, Ang II induced a significantly greater increase in blood pressure in males (delta 37.8 mmHg, 107.2mmHg to 139.5mmHg, p<.05) while female mice had similar changes as seen in controls (delta 16.7 mmHg, 110.1mmHg to126.8mmHg). Flow cytometric analysis of CD3+, CD3+CD4+, CD3+CD4+Foxp3 and CD3+CD8+, lymphocyte surface markers from Rag-1-/- males and females was performed on whole blood, splenic, renal and brain tissue. Absolute CD3+ T cell count from spleen suggested that both sexes had equal T cells engraftment. Males had significantly greater numbers of CD3+CD4+, CD3+CD4+Foxp3 and CD3+CD8+ lymphocyte infiltration of the kidney compared to females in both control and AngII infused animals (p<0.05 comparison of males to females). Males also had significantly increased CD3+CD4+ brain infiltration relative to females. These results suggest that intact female Rag-1-/- mice during Ang II infusion are protected from the hypertensive effects of CD3+ male T cell adoptive transfer as compared to their male counterparts and this protection may involve sex differences in T cell infiltration of the kidney and brain.
Abstract 516: Urinary Proteomic Biomarkers to Predict Cardiovascular Events
Authors: Delles Christian , Brown Catriona E , McCarthy Nina S , Albalat Amaya , Mullen William , Sattar Naveed , Hughes Alun D , Thom Simon , Mayet Jamil , Stanton Alice , Sever Peter S , Dominiczak Anna F , Mischak Harald
Published: March 2022

We have previously demonstrated associations between the urinary proteome profile and coronary artery disease (CAD) in cross-sectional studies. Here we evaluated the potential of urinary proteomics as a predictor of CAD in the Hypertension Associated Cardiovascular Disease (HACVD) sub-study population of the ASCOT study.

Thirty-seven cases with the primary endpoint CAD (fatal CAD, non-fatal myocardial infarction and coronary revascularisation) but without established cardiovascular disease at baseline were selected and matched for sex and age within ±2 years to controls who had not reached a CAD endpoint during the study (median observation time, 5 years). A spot urine sample collected at 1 to 1.5 years post randomisation was analysed using capillary electrophoresis (CE) on-line coupled to Micro-TOF mass spectrometry (MS). A previously developed 238-marker CE-MS model for diagnosis of CAD (CAD
238
) was assessed for its predictive potential.

Sixty urine samples (32 cases; 28 controls; 88% male, mean age 64±5 years) passed quality control for proteomic analysis. There was a trend towards lower ("healthier") values in controls for the CAD model classifier (-0.432±0.326
vs
-0.587±0.297,
P
=0.062), and the CAD model showed statistical significance on Kaplan-Meier survival analysis (Log Rank (Mantel-Cox)
P
=0.021). After unblinding we found 190 individual markers out of 1501 urinary peptides that separated cases and controls with an AUC>0.6. Of these, 28 peptides including fragments of PGRC1, PTGDS, CO1A1, CO3A1, COGA1, PXDC2, B3GT6 and RET4 were also components of CAD
238
.

A urinary proteome panel that was originally developed in a cross-sectional study predicts CAD endpoints independent of age and sex in a well controlled prospective study. Proteomic analysis may have the potential to detect subclinical cardiovascular damage that is associated with increased cardiovascular risk.
Abstract 518: Systems Biology Approach to Identify Genetic Mechanisms Underlying the Metabolic Syndrome in the LH Rat
Authors: Wang Jinkai , Ma MC John , Mennie Amanda K , Pettus Janette , Xing Yi , Kwitek Anne E
Published: March 2022

The metabolic syndrome (MetS) is a collection of co-occurring complex disorders including hypertension, obesity, dyslipidemia, and insulin resistance. The Lyon Hypertensive (LH) and Lyon Normotensive (LN) rats are models of MetS sensitivity and resistance, respectively. To identify genetic determinants and mechanisms underlying MetS, rats from an F2 intercross between LH and LN were studied. Multi-dimensional data were obtained including SNP genotypes, physiological traits including blood pressure, plasma lipid and leptin levels, and body weight/adiposity, and more than 150 billion nucleotides of RNA-Seq reads from the livers of F2 (n=36), LH (n=6), and LN (n=6) individuals. We identified 17 pQTLs (physiological quantitative trait loci) and 1200 eQTLs (gene expression quantitative trait loci) at genome-wide p<0.05. Systems biology methods were applied to prioritize candidate MetS genes, including genes previously shown to be MetS-related. We identified an eQTL peak on rat chromosome 17 (RNO17) shared by large number of genes; RNO17 also has pQTL for MetS-related traits. Network analysis in these genes identified two major co-regulation modules involving mitochondria and gene regulation; genes in the two modules causally affect multiple MetS traits. A gene,
RGD1562963,
closest to the eQTL peak was identified as the eQTL driver gene. It is directly affected by genetic variation between LH and LN rats, affects downstream genes in the eQTL cluster, and is correlated with MetS phenotypes. Our study sheds light on the intricate pathogenesis of MetS and proved that systems biology with high-throughput sequencing is a powerful method to study the etiology of complicated diseases.
Abstract 521: Upregulation of Fibroblast Growth Factor 23 Contributes to Klotho Gene Deficiency-induced Hypertension
Authors: Schuman Mariano , Sun Zhongjie
Published: March 2022

Objective.
Klotho is a newly-discovered anti-aging gene. Klotho (KL) gene delivery prevented progression of spontaneous hypertension and attenuated kidney damage. The purpose of this study is to determine if KL deficiency affects blood pressure and investigate the underlying mechanism.

Methods & Results.
Two groups of wild-type (WT) and 2 groups of KL heterozygous (+/-) mice were used. Blood pressure was elevated significantly in KL(+/-) mice
vs
the WT mice (mean BP 126
vs
103 mmHg), suggesting that KL gene deficiency causes hypertension. Plasma level of FGF23 was increased significantly in the KL(+/-) mice. We then inhibited FGF23 by a FGF receptor inhibitor (PD173074). One group of each strain received PD at 1 mg/kg/day (IP) for one week followed by 10 mg/kg for another week, while the other group received vehicle and served as a control. Following the 2-week treatment, the mice were euthanized and blood, kidney, heart were collected. Distal convoluted tubules (DCT) were isolated from 1 kidney of each animal. PD significantly attenuated hypertension and cardiac hypertrophy. Interestingly, the level of urine norepinephrine was increased in KL(+/-) mice (KL(+/-) 490 ng/ml
vs
WT 350 ng/ml), suggesting that KL deficiency activates the sympathetic nervous system (SNS). PD abolished the KL deficiency-induced increase in urine norepinephrine.

Conclusions.
Klotho deficiency causes hypertension and cardiac hypertrophy probably
via
upregulation of FGF23 which further activates the SNS.
Abstract 524: Vinegar Enhanced the Anti-hypertensive Effect of Saccharina Japonica Intake in Rats With 2-kidney, 1-clip Renovascular Hypertension
Authors: Kitamura Saori , Hashimoto Hiroko , Segawa Yukiko , Kurihara Nobutaka
Published: March 2022

Objective:
Saccharina japonica (SJ) is one of brown algae and is cultivated or grows wild in Japan and neighbor countries. SJ is rich in alginate, the intake of which was reported to decrease blood pressure (BP) in spontaneously hypertensive rats. It is also rich in fucoidan, which was reported to improve the renal blood flow in rats with acute renal failure. We recently observed that intake of SJ decreased BP in 2-kidney, 1-clip renovascular hypertensive (2K1C) rats, probably through nitric oxide, and did not change BP in normotensive rats. In Japan, there are many traditional processed foods made of SJ with vinegar, which is originally used for softening SJ and raising its preserving property. Vinegar is also suggested to suppress angiotensin-converting enzyme activity. In this study, we investigated the interactive effects of SJ and vinegar on BP in 2K1C rats.

Methods:
Male Sprague-Dawley rats (6 wks) were treated with sham operation (SHAM) or clipping the left renal artery (2K1C). After surgery, SHAM or 2K1C rats started receiving a control diet (CTL) or a diet containing 5.0% (w/w) freeze-dried SJ powder (SJ) for 6 weeks, and a tap water (Veh) or a water containing 5.0% (v/v) of 4.3% acidity rice vinegar (VG). The systolic BP (SBP) was measured by a tail-cuff method every week. At the end of the protocol, mean arterial BP (MAP) was measured in each rat under anesthesia. Then, thoracic aorta was removed and the section of aorta was stained with hematoxylin and eosin.

Results:
Analysis of variance revealed significant effects in time, animal model (SHAM
vs
2K1C), diet (CTL
vs
SJ) and model x diet (
P
< 0.001, each), but not in water (Veh
vs
VG) or model x water. At the end of the protocol, in CTL-Veh, 2K1C was higher in SBP than SHAM (179 ± 33
vs
130 ± 20 mmHg,
P
< 0.001). In 2K1C-Veh, the SJ diet decreased SBP (156 ± 6 mmHg) compared with CTL (
P
< 0.05). Vinegar did not significantly change SBP (184 ± 19 mmHg) in 2K1C-CTL, but decreased SBP (142 ± 11 mmHg) in 2K1C-SJ (
P
< 0.05) as low as in SHAM-SJ-VG (139 ± 3 mmHg,
P
= NS). The observations in MAP were similar to in the SBP. Moreover, compared to SHAM, 2K1C-CTL-Veh showed the aortic media thickening, which was reduced in 2K1C-SJ-Veh/VG.

Conclusion:
A certain daily intake of vinegar did not lower BP by itself, but enhanced the BP reduction by a SJ diet, in 2K1C rats.
Abstract 525: Evaluation the Effect of Green Tea on Blood Pressure, Endothelial Function, Metabolic Profile, Inflammatory Activity and Body Adiposity in Obese Pre-hypertensive Women
Authors: Nogueira Lívia d , Rodrigues Maria d , Valença Débora C , Torres Márcia R , Neto José F , Marques Ruy G , Soares Elaine , Guedes Marcella , Sanjuliani Antonio F
Published: March 2022

Background:
Cardiovascular diseases are the leading cause of mortality in Western countries. Some studies have suggested that green tea has beneficial effects on different cardiovascular risk factors. However, others have failed to show such an association.

Objective:
To evaluate the effects of green tea on blood pressure, endothelial function, metabolic profile, inflammatory biomarkers and body adiposity in obese pre-hypertensive women.

Methods:
Crossover randomized controlled double-blinded trial. Twenty women with obesity and pre-hypertension, aged 28-59 years, with stable body weight were randomized to receive a daily supplement of 3 capsules that contained either 500mg of green tea extract (GTE) or a matching placebo for 4 weeks, with a washout period of 2 weeks between the treatments. Blood pressure was evaluated by ambulatory blood pressure monitoring (ABMP). Endothelial function was evaluated by peripheral arterial tonometry method, using Endo-PAT 2000®. The inflammatory status was assessed by interleukin 6, tumor necrosis factor alpha, vascular cell adhesion molecule, intercellular adhesion molecule, plasminogen activator inhibitor-1, vascular endothelial growth factor, E-selectin, adiponectin and C-reactive protein.

Results:
After administration of GTE compared with placebo there was a significant reduction in systolic blood pressure during 24h (pre 130.3±1.7 vs. post 127.0±2.0 mmHg; p= 0.02), daytime (pre 134.0±1.7 vs. post 130.7±2.0 mmHg; p= 0.04) and nighttime (pre 122.2±1.8 vs. post 118.4±2.2 mmHg; p= 0.02). After consumption of green tea, there was an increase, although not statistically significant, in reactive hyperemia index (pre 1.98±0.10 vs. post 2.22±0.14), besides reduction in the concentration of intercellular adhesion molecule (pre 91.8±8.0 vs. post 85.8±5.6 ng/ml) and vascular endothelial growth factor (pre 195.8±46.2 vs. post 158.6±38.7 pg/ml), however without statistical significance. The other variables did not change significantly after consumption of green tea.

Conclusions:
The findings of the present study suggest that green tea has a beneficial effect on blood pressure and possibly on endothelial function.
Abstract 538: Prolonged Functional Sympathetic Efferent Denervation Following Ablation of the Renal Nerves in SHR
Authors: Denton Kate M , Tjongue Melissa , Tare Marianne
Published: March 2022

Renal denervation (RDN) has been shown to be an effective and safe treatment for human resistant hypertension and is considered one of the most promising advances in the field of hypertension. However, the mechanisms mediating the sustained decrease in arterial pressure in response to RDN in humans are unknown. This issue is particularly perplexing, given reports that the depressor effect of RDN in animal models is short lived. The current dogma in the literature is that overtime the efferent sympathetic nerves regrow and so the longevity of the depressor effect of RDN in humans cannot be ascribed to the absence of efferent renal sympathetic nerve activity. We hypothesize that the renal sympathetic nerves regenerate following RDN, and arrive at the target tissues, but the functional response is not fully restored and this contributes to the long-term depressor effect of RDN. Our aim was to examine mean arterial pressure (MAP) and vascular function in spontaneously hypertensive rats (SHR) following RDN. In 10 week old male SHR radiotelemetry probes were implanted to measure MAP. At 12 weeks of age, following 3 days basal MAP recording, sham (n=5) or bilateral RDN (n=5) surgery was performed. MAP was recorded until 24 weeks of age (12 weeks post-RDN). Renal lobar arteries were mounted on a myograph and membrane potential and tension responses to perivascular nerve stimulation assessed. Perivascular nerve stimulation with single pulses at increasing voltage evoked excitatory junction potentials (Ejps) of increasing amplitude in SHR-sham. Ejp amplitude was markedly reduced in arteries from SHR-RDN (P<0.01). Repetitive stimulation (1-8Hz, 5s) evoked slow depolarisation and contraction. At 5Hz 5s, slow depolarisation was 12±2 mV in SHR-shams and 7±2 mV in SHR-RDN (P<0.01), and contractions were smaller than SHR-sham (10 ± 3% vs 22 ± 2% HiK; P<0.01). In conclusion, smaller Ejps suggest that close contacting neuroeffector junctions are not reformed and blunted vasoconstrictor responses indicate neuro-vascular function has only partially returned 12 weeks post-RDN (Fig. 2). Our finding of continued ‘functional’ denervation is compatible with our data that RDN causes a sustained decrease in arterial pressure in SHR.
Abstract 54: Diminished Endothelial Fibrinolytic Function With Prehypertension
Authors: Diehl Kyle J , Weil Brian R , Greiner Jared J , Stauffer Brian L , DeSouza Christopher A
Published: March 2022

Prehypertension (systolic blood pressure: 120-139 mm Hg and/or diastolic blood pressure: 80-89 mm Hg) affects ~30% of the US adult population and is associated with increased atherothrombotic vascular disease risk. The underlying mechanisms responsible for this heightened risk are unclear. The capacity of the endothelium to release tissue-type plasminogen activator (t-PA), the primary activator of the fibrinolytic system, is an important endogenous defense mechanism against intravascular fibrin deposition and thrombosis. As part of an ongoing study, we are determining whether the capacity of the endothelium to release t-PA is impaired in adults with prehypertension. To date, 22 sedentary, non-obese middle-aged men have been studied: 12 normotensive (age: 53±2 yr; BP: 113/72±2/2 mm Hg) and 10 prehypertensive (age: 55±2 yr; BP: 133/85±2/1 mm Hg). All subjects were free of overt cardiometabolic disease. Net endothelial release of t-PA was determined, in vivo, in response to intrabrachial infusions of bradykinin (BK: 125-500 ng/min) and sodium nitroprusside (SNP: 2-8 μg/min). Endothelial t-PA release in response to each dose of BK was significantly lower (~50%) in the prehypertensive (from -1.7±0.7 to 34.1±5.7 ng/100 mL tissue/min) vs the normotensive (from 0.6±1.4 to 54.1±5.7 ng/100 mL tissue/min) adults. Total amount of t-PA released (area under the BK curve) was ~40% less (P<0.05) in the prehypertensive compared with normotensive group (210±32 vs 336±45 ng/100 mL tissue). There was no effect of SNP on t-PA release in either group. In the overall study population, there was a significant inverse relation between both systolic (r=-0.44) and mean arterial (r=-0.42) blood pressure and total t-PA release. No other anthropometric, hemodynamic or metabolic variable was associated with t-PA release. These initial results indicate that the capacity of the endothelium to release t-PA is impaired in prehypertensive men. Diminished endothelial fibrinolytic capacity may contribute to the increased atherothrombotic risk with prehypertension.
Abstract 544: Development of a Stabilized Bioactive VEGF Chimera for the Management of Preeclampsia
Authors: George Eric , Liu Huiling , Bidwell Gene L
Published: March 2022

Preeclampsia is a gestational hypertensive disorder which occurs in 5-10% of all pregnancies in the US. While the underlying cause of the disease is unknown, there is growing recognition that placental ischemia leads to the secretion of pathogenic factors into the maternal blood stream, perhaps most notably the VEGF antagonist sFlt-1. Circulating sFlt-1 sequesters free VEGF and decreases the overall level of VEGF available for cell signaling. Preclinical experiments suggested a palliative effect of VEGF delivery in preeclampsia models, but translation of this therapy is severely hampered by the short half-life of free VEGF and convenient routes of administration. We have recently constructed a VEGF chimera fused to an Elastin Like Polypeptide (ELP) polymer with significantly increased
in vivo
half-life and stability which appears to retain full VEGF signaling activity. Importantly, we have preliminary evidence that the ELP fusion prevents placental translocation and fetal exposure, preventing harmful fetal effects. ELP-VEGF stimulated normalized HUVEC proliferation at 72hr was dose dependent from 1-100nM, and was equivalent to that of free VEGF (2 ± 0.25 for ELP-VEGF vs 1.94 ± 0.18 at 100nM, p<0.05 versus control for both). Free VEGF (20nM) induced tube formation by HUVECs (8.8 ± 2 vs 14.5 ± 3 tubes/field, p<0.05). Likewise ELP-VEGF significantly increased tube formation compared to control cells (p<0.05), and trended toward augmentation of tube formation versus VEGF alone (18.6 ±4 tubes per frame, p=0.07), though this did not reach significance. Finally, ELP-VEGF (50nM) stimulated HUVEC migration through matrigel (9.9 ± 0.7 vs 3.4 ± 0.3 cells/field, p<0.05) to a similar extent to free VEGF (8.4 ± 0.6 cells/field, p<0.05 vs control). Together, these data suggest that ELP-VEGF retains full VEGF signaling potential, which along with enhanced pharmacokinetic parameters could be a useful therapeutic for the restoration of VEGF signaling in the preeclamptic patient.
Abstract 545: PLacental eXpanded (PLX) Cell Treatment Ameliorates Preeclampsia Induced by TLR3 or TLR7 Activation in Mice
Authors: Chiasson Valorie L , Chatterjee Piyali , Hatahet Mohamad , Pinzur Lena , Abraham Eytan , Flaishon Liat , Chajut Ayelet , Mitchell Brett M
Published: March 2022

PLacental eXpanded (PLX) cells (Pluristem Therapeutics Inc.) are human placenta-derived, mesenchymal-like adherent stromal cells that release proteins in response to the environment of the host. PLX cells are non-immunogenic and have been shown to decrease inflammation and increase angiogenesis in inflammatory and ischemic conditions. Therefore, we tested whether PLX cell treatment could attenuate symptoms of preeclampsia (PE) in mice. We hypothesized that one-time PLX cell treatment would decrease the pregnancy-dependent hypertension, proteinuria, endothelial dysfunction, splenomegaly, inflammation, and placental injury induced by Toll-like receptor (TLR) activation during pregnancy. Pregnant C57BL/6 mice were given ip injections of saline vehicle (P), the TLR3 agonist poly I:C (PPIC), or the TLR7 agonist R837 (PR) on days 13, 15, and 17 of gestation. P, PPIC, and PR mice were also given either plasmalyte A (PLA, vehicle) or PLX cells (1 million) by im injection in the right leg on gestational day 14 (n=8 in each group). PLX cell treatment progressively decreased SBP over 3 days in PPIC and PR mice and had no effect in P control mice (day 17 SBP in mmHg: P+PLA = 100±4, P+PLX = 96±4, PPIC+PLA = 144±3, PPIC+PLX = 111±1, PR+PLA = 145±2, PR+PLX = 106±3; PPIC+PLA and PR+PLA p<0.05 vs. P+PLA). PLX cell treatment also normalized the urinary protein/creatinine ratio and aortic endothelium-dependent relaxation responses in PPIC and PR mice to that of P mice while having no significant effects on the number of fetuses or incidence of fetal demise per litter. Inflammation plays a central role in the development of TLR-induced PE and PLX cell treatment reduced spleen weight/body weight ratios, normalized splenic levels of gamma-delta T cells, decreased plasma IL-6 levels, and restored plasma IL-4 levels in PPIC and PR mice. Additionally, PLX cell treatment reduced fibrin deposition in the placental vasculature and significantly reduced placental HIF-1alpha protein levels. These data demonstrate that one-time PLX cell treatment after PE is induced was able to decrease inflammation, proteinuric hypertension, endothelial dysfunction, and placental injury in mice and may be beneficial in women with PE.
Abstract 55: PPAR-δ Binding to Heme Oxygenase 1 Promoter Prevents Angiotensin II Induced Vascular and Adipocyte Dysfunction in a Model of Renovascular Hypertension
Authors: Sodhi Komal , Puri Nitin , Khitan Zeid , Dial Larry , Shapiro Joseph , Abraham Nader G
Published: March 2022

Introduction:
Renin-Angiotensin System (RAS) induces oxidative stress and contributes to various pathological conditions including insulin resistance and the metabolic syndrome. Recent studies have shown the role of PPARδ-agonist in attenuation of Angiotensin II induced oxidative stress.The aim of this study was to explore the effectiveness of a PPARδ-agonist in the prevention of Ang II-induced vascular and adipocyte dysfunction and the possible interaction between PPARδ and HO-1 system in a model of enhanced oxidative stress, the Goldblatt 2K1C model.

Methods:
We first established a direct stimulatory effect of the PPARδ-agonist (GW 501516) on the HO-1 gene by demonstrating increased luciferase activity in COS-7 cells transfected with a luciferase-HO-1 promoter construct. SD rats were divided in 4 groups: sham operated animals, 2K1C rats and 2K1C rats treated with GW 501516, in the absence or presence of the HO activity inhibitor, stannous mesoporphyrin (SnMP).

Results:
2K1C animals had increased blood pressure, visceral adiposity, adipocyte hypertrophy, increased inflamatory cytokines, increased circulatory and adipose tisssue levels of renin and Ang II along with increased adipose tissue gp91 phox expression (p<0.05) when compared to sham operated animals. Treatment with GW 501516 increased adipose tissue HO-1 and adiponectin levels (p<0.01) along with enhancement of Wnt10b and β-catenin expression. HO-1 induction was accompanied by the decreased expression of Wnt 5b, Mest and C/EBPα levels and an increased number of small adipocytes (p<0.05). These effects of
GW501516
were reversed in 2K1C animals exposed to SnMP (p<0.05).

Conclusion:
This novel study corroborates the existence of a PPARδ-HO-1 axis whose direct stimulation via the administration of an exogenous PPARδ-agonist induces HO-1 expression, abates RAS associated adipocyte and vascular dysfunction and induces adipocyte remodeling with smaller adipocytes while decreasing inflammation and increasing adiponectin secretion.