bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P235: Reduced Uterine Perfusion Pressure Impairs Seizure-induced Downregulation Of Neurotransmitter Transporters In Mice
Authors: Jones- Muhammad Maria , Stancil Kennedi , Shao Qingmei , Warrington Junie P
Published: August 2021

Preeclampsia, a hypertensive disorder of pregnancy, can advance to eclampsia, if new onsetseizures occur. Previous work showed increased susceptibility to chemically-induced seizures inthe reduced uterine perfusion pressure (RUPP) rat model of preeclampsia; however, theunderlying mechanisms are unknown. Because seizures occur due to neurotransmitter activityimbalance, we hypothesized that RUPP mice have elevated excitatory and reduced inhibitoryactivity and that seizures exacerbate this imbalance.Timed-pregnant SMA-GFP mice (n=5-6 per group/treatment) were subjected to sham or RUPPsurgery on gestational day (GD) 13.5 and seizures were induced using 40mg/kg pentylenetetrazolon GD18.5. Tissues were harvested 30 minutes post-seizure induction. Maximum seizure scoreswere similar in sham (4.7±0.3) and RUPP (4.5±0.3) mice; p=0.37. Fluorometric assay showsseizures increased [F (1, 16) = 5.99, p=0.03], while RUPP had no effect [F (1, 16) = 1.15, p=0.3]on hippocampal glutamate concentration. No pairwise differences were observed within the shamand RUPP groups exposed to seizures (p>0.05). Seizures increased [F (1, 16) = 6.96, p=0.02],while RUPP had no effect [F (1, 16) = 0.61, p=0.45] on GABA concentration, with no significantpairwise difference in GABA concentration (p>0.05).Western blot analysis shows seizures significantly reduced hippocampal NMDAR1 (1.0±0.5 vs0.6±0.96, p=0.02; 1.0±0.1 vs 0.6±0.0, p=0.04) and GABAAR receptor expression (1.0±0.4 vs0.35±0.1, p<0.5; 1.05±0.3 vs 0.3±0.1, p<0.05) in sham and RUPP mice. Following seizureexposure, vesicular glutamate transporter (VGLUT1: sham: 1.0±0.3 vs 0.5±0.1; p<0.01, RUPP:0.8±0.2 vs 0.6±0.1; p=0.11), excitatory amino acid transporter 1 (EAAT1: sham: 1.0±0.4 vs0.6±0.2; p=0.02, RUPP: 0.9±0.1 vs 0.6±0.2; p=0.17) and GABA transporter (GAT1: sham: 1.0±0.5vs 0.4±0.1, p=0.01; RUPP: 0.8±0.2 vs 0.5±0.1, p=0.12) was reduced in sham mice, but not RUPPmice.Although RUPP does not change baseline GABA or glutamate related receptor or transporterexpression, our findings suggest seizure-induced reductions in vesicular and astrocyticneurotransmitter transporters is impaired by the RUPP procedure. Ongoing studies assesswhether other receptors and transporters are affected.
Abstract P237:
Alterations In Renal Sodium Handling Proteins Determined By Urinary Extracellular Vesicle Analysis During Dietary Sodium Modulation And In Primary Aldosteronism
Authors: Luther James M , Peng Dungeng , Vaidya Anand , McDonald Hayes , Schey Kevin , Adler Gail K
Published: August 2021

Sodium handling is altered during extremes in dietary sodium intake or in pathologic conditions such as primary aldosteronism. Urinary extracellular vesicles (uEVs) are excreted by all cell types along the nephron, are enriched for membrane proteins, and could provide insight into renal physiology in humans. We assessed uEV protein excretion of renal epithelial proteins relevant to sodium handling from 54 volunteers with pre-hypertension on high (HS, 200 mmol/d) and low (LS, 10 mmol/d) sodium diet for 5-7 days. To compare pathologic versus physiologic responses to aldosterone excess, samples were collected from 9 patients with primary aldosteronism (PA). uEVs were isolated by ultracentrifugation and trypsin digested proteins were quantified using multiple reaction monitoring mass spectrometry using isotopically labeled standards, normalized to the uEV-specific marker CD9. Wilcoxon signed rank and rank sum tests were used for within-subject and between-subject comparisons, respectively. LS diet altered uEV excretion of pendrin (4.0, expressed as fold-change vs HS), aquaporin-2 (-2.03), TRPV5 (-1.91), NCC (1.51), OSR1 (1.42;
P
<0.01 for all), and SGLT2 (1.20,
P
=0.042). In primary aldosteronism, expression of ENaC-α, ENaC-β, and ENaC-γ, NCC, SGLT2, and OSR-1 were increased to a greater extent compared to during LS diet (
P
<0.05 for each), whereas AQP2 and TRPV5 were similar to HS diet (
Figure
). Expression of uEV proteins relevant to renal tubular sodium handling are dynamically altered by dietary sodium restriction and aldosterone excess. Further studies could identify patterns specific to pathologic versus physiologic aldosterone responses.
Abstract P246: Relationship Between Urinary Sodium/potassium Ratio And Systolic Blood Pressure In Salt Sensitive Subjects In The Dash-sodium Trial
Authors: Abramov Rozalia , Drugge Elizabeth D , Farhan Khalid A , Ferreri Nicholas R
Published: August 2021

Excessive salt intake is associated with hypertension and cardiovascular morbidity. However, identifying those at risk of salt sensitive hypertension (SSH) remains a challenge due to its unequal distribution among populations and inaccurate assessment of dietary sodium (Na) and potassium (K) intake. The objective of this study was to compare indices of dietary Na intake in relation to systolic blood pressure (SBP) in salt sensitive (SS) and salt resistant (SR) subjects from the Dietary Approaches to Stop Hypertension (DASH)-Sodium trial. We hypothesized that when compared to urinary Na or K independently, Na/K ratio is a better predictor of SSH when defined as a 5-10 mmHg change in SBP from low to high dietary Na. Among 404 Black and White subjects, baseline classifications included 177 SS and 227 SR. After diet randomization, on the control 107 were SS and 92 SR and on the DASH 70 were SS and 135 SR. Descriptive statistics, bivariate analysis, followed by linear regression models for baseline and multilevel mixed-effects models after intervention were used to assess the relationship between SBP and dietary Na (as measured by urinary Na/K ratio or Na and K independently) using SS as a categorical factor. SBP was consistently associated with SS, Na/K ratio, and age in all models
.
At baseline, SBP was significantly higher in SS and SR subjects of the same race and sex, after controlling for age and urinary Na/K ratio and was highest for White females, SS:142.3 (138.8, 145.7) vs. SR:133.2 (130.7, 135.7), and for Black males, SS:137.0 (133.8, 140.1) vs. SR: 129.6 (127.0, 132.3). On average, SBP increased 1.02 (0.065, 1.98) mmHg with each unit increase in Na/K ratio and 3.30 (2.41, 4.19) mmHg with each 10-year increase in age. After randomization and exposure to increasing levels of sodium, SBP increased in SS subjects on the control diet:125.3 (123.2, 127.3) to 136.8 (134.8, 138.9), an effect that was greater in White vs Black females and in Black vs White males. SBP increased in SS subjects on the DASH diet: 121.4 (118.8, 124.0) to 131.2 (128.7, 133.7), but there were no differences by race and sex. These results suggest that a 5-10 mmHg change in SBP in subjects on a typical American diet and Na/K ratio are good predictors of SSH and that the DASH diet may help to reduce race and sex disparities.
Abstract P247: Elastin Insufficiency Impairs Renal Hemodynamics And Accelerates Aging-associated Structural And Functional Remodeling Of Preglomerular Arterioles
Authors: Dixon Alethia , Osei-Owusu Patrick
Published: August 2021

Elastin degradation and fragmentation are hallmarks of arterial stiffness and renal dysfunction associated with aging. However, it is unclear whether elastin insufficiency contributes to the changes in the structure and function of the resistance vasculature of the kidney during aging. Here we determined how increased vascular stiffness due to elastin insufficiency alters renal hemodynamics and mechanical properties of preglomerular arterioles. We assessed renal hemodynamics under anesthesia in 14-16-month-old female wild type (WT) and elastin heterozygous (
Eln

+/-

) mice. Renal autoregulation was assessed by a stepwise increase in renal perfusion pressure (RPP) by simultaneously occluding the superior mesenteric and celiac arteries. Myogenic constriction and arterial stiffness were assessed by pressure myography of isolated renal interlobar arteries. Baseline renal vascular resistance (RVR) was elevated in
Eln

+/-

mice (13.8 ± 2.9 vs 11.2 ± 1.4 mmHg/μL/min/g left kidney weight), while systolic blood pressure (SBP; 75.1 ± 7.4 vs 91 ± 4.2mmHg), renal blood flow (RBF; 6.3 ± 1.2 vs 7.4 ± 1.7 μL/min/g left kidney weight), renal plasma flow (RPF; 3.4 ± 0.8 vs 5 ± 1.2 mL/min/g/ left kidney weight) and urine flow rate, all trended lower in
Eln

+/-

mice compared to WT mice. Glomerular filtration rate (GFR) and filtration fraction (FF) were similar between the two groups. A stepwise increase in RPP caused a slower decline and rise in RBF and RVR, respectively, in
Eln

+/-

relative to WT mice. The maximal changes in RBF (5 ± 1.1 vs 4.7 ± 0.8 μL/min/g left kidney weight), RVR (17.6 ± 7.3 vs 22.5 ± 2.1 mmHg/μL/min/g left kidney weight), urine flow rate,GFR, and FF were less robust in Eln
+/-
mice. RPF decreased in WT mice in response to raising RPP, whereas it remained unchanged in
Eln

+/-

mice. Myogenic response and increases in elastic modulus and wall tension following stepwise changes in intraluminal pressure were all augmented in interlobar arteries from
Eln

+/-

relative to WT mice. However, there was no difference in kidney weight/tibia length ratio between the two genotypes. We conclude that elastin insufficiency impairs renal hemodynamics by exacerbating age associated increase in vascular stiffness.
Abstract P251: Out-of-clinic Cortisol Secretion Is Increased In Masked Uncontrolled Hypertension
Authors: Siddiqui Mohammed , Carey Robert M , Calhoun David A , Oparil Suzanne
Published: August 2021

Introduction:
Masked uncontrolled hypertension (MUCH) in treated patients is defined as controlled office BP but uncontrolled out-of-clinic ambulatory BP (ABP). In contrast, patients with true controlled hypertensive (CHTN) have controlled BP by both office BP and out-of-clinic ABP. Previously, we have shown that patients with MUCH have increased out-of-clinic catecholamines/metanephrines (indicative of SNS activity) and aldosterone levels (indicative of RAAS activity). The current study tested the hypothesis that MUCH patients have higher cortisol levels (corticotropin-glucocorticoid axis) compared to patients with true CHTN.

Methods:
222 patients with controlled office BP at ≥3 clinic visits were recruited. Patients taking glucocorticoid-containing medications were excluded. All patients were evaluated by automated office BP. Out-of-clinic ABP monitoring and 24-hour-urinary cortisol, cortisone, aldosterone, catecholamines and metanephrines levels were also measured. 115 patients had MUCH and remaining 75 patients had true CHTN.

Results:
MUCH patients had significantly higher out-of-clinic levels of 24-hour-urinary cortisol, catecholamines, metanephrines and aldosterone compared to true CHTN. Further, in correlation matrix analysis higher urinary cortisol was associated with higher catecholamines, metanephrines and aldosterone in MUCH patients.BP.

Conclusions:
Patients with MUCH had higher out-of-clinic urinary cortisol levels compared to patients with true CHTN. These findings suggest that patients with MUCH have higher out-of-clinic cortisol, aldosterone secretion and SNS tone that may contribute to their higher out-of-clinic BP.
Abstract P252: Smooth Muscle At1a Receptor Mediates Perivascular Fibrosis In Angiotensin Ii-infused Mice
Authors: Okuno Keisuke , Eguchi Satoru , Sparks Matthew A
Published: August 2021

Angiotensin II (Ang II) signaling via AT1 receptor has been shown to play a critical role in the pathogenesis of hypertension, cardiovascular hypertrophy and fibrosis. We have demonstrated that ADAM17 expressed in vascular smooth muscle cells (VSMC) mediates EGF receptor activation and promotes cardiac hypertrophy and perivascular fibrosis induced by Ang II. It is conceivable that Ang II signaling in VSMCs specifically initiates cardiovascular remodeling, such as hypertrophy and fibrosis. In a recent study, deficiency of smooth muscle AT1a receptors results in diminished hypertension and protection from cardiac hypertrophy induced by Ang II. However, we have limited understanding whether smooth muscle AT1a receptors affects hypertensive fibrosis in vasculature. Thus, this study was designed to elucidate the roles of the AT1a receptor in VSMCs in cardiovascular remodeling including fibrosis during Ang II stimulation using VSMC AT1a receptor deficient mice. To delete the AT1a receptor from VSMCs, we crossed C57BL/6 transgenic mouse lines expressing Cre recombinase under the control of the sm22α promoter (KIsm22α-Cre). Male AT1a
flox/flox
KIsm22α-Cre+/- (SMKO) and Controls (AT1a
flox/flox
KIsm22α-Cre-/-) mice were infused with Ang II (1 μg/kg/min) for 2 weeks via osmotic mini-pump. In Control mice, Ang II infusion for 2 weeks induced cardiac hypertrophy indicated by heart-to-body weight ratio and echocardiogram. After 2 weeks of Ang II infusion, heart-to-body weight ratios were significantly increased in Control mice compared with AT1a SMKO mice (6.04 versus 4.89, respectively, p=0.032). Cardiac wall hypertrophy was seen in Controls after 2 weeks of Ang II infusion, which was attenuated in AT1a SMKOs. Control mice (n=5) showed vascular medial hypertrophy and perivascular fibrosis, whereas these phenotypic changes were attenuated in SMKO mice (n=4). In conclusion, AT1a receptors from VSMC could mediate Ang II-induced cardiovascular hypertrophy and perivascular fibrosis. Whether the data can be fully explained by the prevention of hypertension remains to be determined, the data contrast to the past manuscript showing a protective effect in AT1a
flox/flox
S100A4-Cre+/- mice (fibroblast silencing) with Ang II infusion.
Abstract P253: Relationship Between Aldosterone And Parathyroid Hormone In Patients With Primary Hyperparathyroidism
Authors: Stanic Branka , Ilincic Branislava , Zeravica Radmila , Milicic-Ivanovski Dragana , Cabarkapa Velibor , Mijovic Romana
Published: August 2021

In primary hyperparathyroidism, an increased risk of developing cardiovascular disease may exist due to increased activity of the renin-angiotensin-aldosterone system. The aim of this study was to evaluate relationship between parathyroid hormone and aldosterone in patients with primary hyperparathyroidism. The study included 48 patients with primary hyperparathyroidism and 30 healthy subjects who matched age and gender to the study group. This study was conducted at the Center for Laboratory medicine, Clinical center of Vojvodina, Novi Sad, Serbia. Compared to the controls, the study group had statistically significantly higher values of aldosterone (p=0.028), total calcium (p=0.01), ionized calcium (p=0.003) and parathyroid hormone (p=0.00). Serum aldosterone and parathyroid hormone levels were correlated positively in patients with primary hyperparathyroidism (r=0.509, p<0.05). A statistically significant positive correlation between renin and parathyroid hormone (r=0.688, p<0.05) and renin and calcium (r=0.673, p<0.05) was determined in hyperparathyroid patients. In the study group 28 patient had hypertension (58.3%) and in a control group only 9 patients had hypertension (30%), which is statistically significant (p=0.028). In multivariate regression analysis, the strongest predictive variable of aldosterone secretion was parathyroid hormone (p=0.011). An independent relationship between parathyroid hormone and aldosterone in patients with primary hyperparathyroidism and the correlation between renin and parathyroid hormone as well as with calcium, indicate not only the direct but also the indirect associations between parathyroid hormone and aldosterone in primary hyperparathyroidism. These findings may represent another possible model of renin-angiotensin-aldosteron-induced organ damage.
Abstract P255: Resistance Hypertension And Achievement Of Target Blood Pressure Level By Continuous Positive Airway Pressure Long Therapy In Patients With Obstructive Sleep Apnea
Authors: Rekovets Oksana , Sirenko Yuriy , Krushynska Nina , Torbas Olena , Kushnir Svitlana , Sirenko Oleksandr , Primak Galina
Published: August 2021

The aim was to assess the arterial stiffness changes in patients with resistant arterial hypertension (AH) and obstructive sleep apnea (OSA) and possibilities of its correction by continuous positive airway pressure (CPAP)-therapy.

Design:
In 10 month follow-up study were included 46 patients with RAH, who were divided into groups: 1-st - patients with RAH and moderate to severe OSA on CPAP (n=21); 2-nd - patients with RAH and moderate to severe OSA without CPAP (n=25). They underwent somnography by dual-channel portable monitor device, office and ambulatory blood pressure monitoring, echocardiography and applanation tonometry. All patients received similar antihypertensive therapy according to 2013 ESH Guidelines for the management of arterial hypertension.

Results:
Patients with RAH and OSA (mean apnea-hypopnea index (AHI) 36.5±2.7 event h-1) in comparison with patients with RAH without OSA (mean AHI 3.4±0.2 event h-1) had significantly higher body mass index (34.2±0.7 vs 31.6±0.7 kg m-2, P<0.05), uric acid level (6.7±0.1 vs 5.6±0.4 mg dl-1, P<0,05)). Patients with RAH and OSA in comparison with patients with RAH without OSA had higher carotid-femoral pulse wave velocity (PWVcf) (12.1±0.5 vs 10.2 m s-1, P<0,05) and central systolic blood pressure (CSBP) (143.8±2.7 vs 136.2±3.4 mm Hg, P<0,05). During 10 months follow-up in patients with RAH and moderate and sever OSA on CPAP-therapy there were significantly decrease of PWVcf (from 12.1±0.5 to 10.5±0.5 m s-1, P<0,05), decrease office systolic blood pressure (from 147.8±3.7 to 136.7±2.8 mm Hg; P<0,05) and diastolic blood pressure (from 96.8±3.5 to 87.0±3.3 mm Hg; P<0,05) with achievement of target levels in 67,2% patients. Central systolic BP decreased (from 143.8±2.7 to 137.7±2.8 mm Hg; P<0,05).

Conclusion:
The combination of therapy continuous positive airway pressure with antihypertensive treatment in patients with resistant arterial hypertension and moderate to severe obstructive sleep apnea improved achievement of target blood pressure, decreased arterial stiffness and decreased central blood pressure.
Abstract P260: SARS-Cov-2 Spike Protein S1-Mediated Endothelial Injury And Pro-Inflammatory State Is Amplified By Dihydrotestosterone And Prevented By Mineralocorticoid Antagonism
Authors: Kumar Nitin , Hunker Kristina , Kanthi Yogendra , Obi Andrea T , Ganesh Santhi
Published: August 2021

Objectives:
Men are disproportionately affected by the coronavirus disease-2019 (COVID-19) and experience higher mortality as compared to women. Endothelial dysfunction has been proposed as a major inciting factor of pro-inflammatory and thrombotic changes in COVID-19 infection. The vascular effects of androgen signaling and inflammatory cytokines in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-mediated endothelial damage are not defined.

Methods:
Here, we determined the effects of SARS-CoV-2 spike protein-mediated endothelial damage under conditions of exposure to androgens and TNF-α. We tested the therapeutic effects of approved drugs: angiotensin receptor blockade by valsartan and mineralocorticoid receptor antagonism by spironolactone. Circulating endothelial injury markers VCAM-1 and E-selectin were measured in the plasma from men and women diagnosed with COVID-19.

Results:
Exposure of endothelial cells (ECs)
in vitro
to androgen dihydrotestosterone (DHT) exacerbated spike protein S1-mediated endothelial injury transcripts for cell adhesion molecules E-selectin, VCAM-1, and ICAM-1 and anti-fibrinolytic PAI-1 (P<0.05), as well as increased THP-1 monocyte adhesion to ECs (P=0.032). Spironolactone dramatically reduced DHT+S1-induced endothelial activation. Pre-treatment of ECs with valsartan did not increase endothelial activation by S1. TNF-α exacerbated S1-induced EC activation and spironolactone abrogated this effect. Analysis of plasma from patients hospitalized with COVID-19 showed concordant higher VCAM-1 in men as compared to women.

Conclusions:
Androgen exposure promoted spike protein-mediated endothelial injury by increasing markers of inflammation and thrombosis. A beneficial therapeutic effect of the FDA-approved drug spironolactone was observed, supporting a rationale for further evaluation as an adjunct treatment in COVID-19.
Abstract P263: Symptomatic Post-Acute SARS-CoV-2-Infection Hypertension Syndrome
Authors: East Lauren , Mahmoud Zainab , Verma Amanda
Published: August 2021

Introduction:
The Post-COVID Cardiology Clinic at Washington University evaluates and treats patients with ongoing cardiovascular symptoms following acute COVID-19 infection. One clinical manifestation seen in the clinic is an increase in blood pressure, with associated symptoms like chest pain. Our investigation aims to describe the increase in blood pressure seen in symptomatic patients presenting to the Post-COVID Cardiology Clinic.

Methods:
The study employed a retrospective cohort design of consecutive adult patients who presented between September 2020 to May 2021 with cardiovascular symptoms following COVID-19 infection. Demographic information, symptoms, vital signs, and follow-up visit data were collected for the patients. To determine a baseline blood pressure, two blood pressure readings from office visits prior to COVID-19 infection were averaged. The blood pressure values were compared between baseline and cardiology office visits using a non-parametric Wilcoxon test for paired data.

Results:
One-hundred patients were included in the cohort (mean age 46.4 years (SD 46.4); 81% (81) female). At the initial visit, there was a significant increase in systolic (median 128 mmHg) and diastolic (median 83.5 mmHg) blood pressure from baseline (systolic median 121.5, p=0.029; diastolic median 76, p<0.001). All patients with an increase in blood pressure reported symptoms like chest pain. In the subset of 36 (36%) patients that have followed up, 35 (97%) patients were prescribed a new anti-hypertensive or required an increased dose of a prior anti-hypertensive at their initial visit. Blood pressures at follow-up were not significantly different from baseline (median systolic delta= 1.0mmHg, diastolic delta= -1.0mmHg; p>0.05), and 83% (30) reported improvement in symptoms.

Conclusions:
Patients presenting with cardiovascular symptoms post-acute COVID-19 show increased blood pressure when compared to blood pressure prior to infection. During subsequent follow-up appointments, patients showed improvement in their blood pressure and symptoms. While the pathophysiology has yet to be determined, it is likely related to the effects of a proinflammatory state, endothelial dysfunction, dysautonomia, or altered effects of the RAAS.
Abstract P270: Covid19 Hypertension In Primary Care
Authors: Moreno Laura
Published: August 2021

One of the symptoms of Sars-cov-2 is hypertension with diastolic dysfunction. Out of 432 patients, 112 had hypertension and 102 of them had diastolic hypertension that helped identify early infection despite having a negative PCR test. Diastolic hypertension led to pulmonary edema. The use of spironolactone, an aldosterone antagonist selective diuretic, at a dose of 25-50mg po q d reduced the effects of hypertension and pulmonary edema in 94/112 of the treated patients. This treatment was added to the regimen the patient was on for previously diagnosed hypertension or initiated due to Sars-cov-2 infection causing hypertension. The key to this medication is in the renin aldosterone system. Uncontrolled aldosterone promotes myocardial fibrosis and is most likely why many people have long haul covid19 cardiac complications. Aldosterone also causes perivascular fibrosis, blocks the myocardial uptake of norepinephrine, and increases plasminogen activator inhibitor levels. In conjunction with angiotensin II, aldosterone causes vascular damage that sends coagulation signals to platelets, endothelial dysfunction, and decreased vascular compliance. Therefore, aldosterone management plays a major role in the development of both hypertension and heart failure in Sars-cov-2 and a key target for therapeutic interventions.
Abstract P271: Delineating Sex-specific Mechanisms Of Impaired Vasoreactivity In Thermoneutrality
Authors: Chun Ji Hye , Henckel Melissa M , Knaub Leslie A , Walker Lori A , Reusch Jane E , Keller Amy C
Published: August 2021

Cardiovascular disease (CVD) is a leading cause of hospitalization and death. CVD is characterized by impaired vasoreactivity and mitochondrial dysfunction. Perivascular adipose tissue (PVAT), considered brown adipose tissue (BAT), surrounds the vasculature and regulates its response. Preliminary data with rats housed at either their thermoneutrality (TN, 30°C) or room temperature (RT, 22°C) showed diminished vasodilation in aorta from TN rats as compared with those from RT rats (10.2% ± 4.0% (0.159 g of vasodilation capacity, starting from maximal force constriction of 1.563 g) versus 64.2% ± 5.3% (0.909 g of 1.417 g, p<0.001). TN-housed rat aorta also showed less mitochondrial respiration with lipid substrates in multiple states (p<0.05). We hypothesize that remodeling of PVAT phenotype from BAT to white adipose tissue (WAT) may alter mitochondrial lipid utilization and cause vasoreactivity dysfunction. To test this, we housed male and female rats at either RT or TN and investigated their own PVAT + aorta or PVAT from the oppositely- housed animals along with each rat’s own aorta for vasoreactivity ex situ. There was diminished vasodilation in all TN animals with PVAT + aorta (29.2% ± 3.8% (0.269 g of 0.923 g) versus 37.6% ± 6.0% (0.255 g of 0.677 g), p<0.02), with only male animals showing a significant effect from PVAT (p<0.001). In aorta of TN-housed animals analyzed with PVAT from RT-housed animals, female vessels showed an increase in vasodilation capacity as compared to controls (56.8% ± 13.6% (0.589 g of 1.037 g) versus 5.2% ± 2.3% (0.028 g of 0.534 g), p<0.001), strongly suggesting that PVAT not only regulates vasoreactivity, but can repair TN-induced diminished dilation in a sex-dependent manner. All animals at TN had significantly less mitochondrial respiration with lipid substrates (p<0.05), with no sex differences. We further observed a significantly greater amount of lipids in PVAT from male TN-housed animals as compared to that in RT-housed animals (p<0.05), consistent with a WAT phenotype. Our data support that TN alters PVAT phenotype in a sex-dependent manner, resulting in dysfunctional vasoreactivity and mitochondrial function. These targets of CVD in both male and female animals are exciting avenues for novel therapeutics.
Abstract P274: Inhibition Of Abnormal Exosome Release Prevents Excessive Aortic Stiffness And Relaxation Dysfunction In Diet-induced Obesity
Authors: Habibi Javad , DeMarco Vincent G , Chen Dongqing , Hulse Jack L. , Whaley-connell Adam , Hill Michael A , Sowers James R , Jia Guanghong
Published: August 2021

Interactions between over-nutrition and abnormal exosome release impact insulin sensitivity and the development of cardiovascular disease (CVD). Recent data have shown that exosomes can be released from various cell types, including adipocytes and vascular cells, and that they exist in body fluids and tissues functioning as mediators of cell-cell communication. However, the specific role of exosomes in diet-induced excessive vascular stiffness and hypertension has not been explored. Accordingly, we hypothesized that abnormal release of exosomes contributes to western diet (WD)- induced aortic stiffening and impaired vascular diastolic relaxation. We further posited that GW4869, an antagonist of neutral sphingomyelinase 2 (nSMase2) which promotes exosome production and release, would prevent WD-induced aortic stiffening and impaired vascular relaxation. Six week-old female C57BL/6L mice were fed a mouse chow (CD) or WD containing excess fat (46%) and fructose (17.5%) for 16 weeks with or without GW4849. To this point, 200 μl of 0.3 mg/mL GW4869 in 0.9% normal saline (60 μg/mouse; 2-2.5 μg/g body weight) was injected intraperitoneally every 48 hours for 12 weeks. 16 weeks of WD induced an increase of aortic stiffness as examined by pulse wave velocity (PWV) and impaired the aortic vasodilation responses to acetylcholine (Ach) and sodium nitroprusside (SNP) (10
-9
-10
-4
mol/L). However, GW4869 treatment prevented the WD-induced excessive aortic stiffness, as well as impairment of endothelium dependent/independent vascular relaxation. There were no significant differences in blood pressure between each group examined by tail cuff blood pressure measurement. These findings support the hypothesis that abnormal release of exosomes play an important role in WD-induced excessive aortic stiffness, impaired vascular relaxation and CVD in diet-induced obesity.
Abstract P275: Deletion Of Endothelial Leptin Receptor Elevates Blood Pressure And Impairs Endothelial-dependent Relaxation Via Upregulation Of Endothelial Glycolytic Enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3)
Authors: Atawia Reem T. , Faulkner Jessica L. , Kennard Simone , Mehta Vinay , Antonova Galina , Fulton David , Patel Vijay S. , Saeed Muhammad I. , Belin de Chantemèle Eric J.
Published: August 2021

The adipokine leptin plays a crucial role in blood pressure (BP) regulation notably by exerting pressor effects centrally via sympatho-activation and depressor effects via direct activation of its receptor (LepR) peripherally resulting in nitric oxide (NO)-mediated vasodilation. However, the predominant effects and cell type responsible for leptin-mediated NO production is not clearly understood. Herein, we examined the effect of selective deletion of LepR in endothelial cells (LepR
EC-/-
, KO) on BP and vascular function. BP recording via radiotelemetry in male KO and WT (LepR
EC+/+
) mice revealed significant increases in diastolic and mean arterial pressure in KO mice (DBP, WT: 90.2±2.1 vs. KO: 100.1±3.6; MAP, WT: 105.7±2.1 vs. KO: 113.7±2.6 mmHg, n=5, p<0.05). There was no difference in Systolic blood pressure or heart rate between KO and WT. Leptin infusion (0.9mg/kg/day,7 days) elicits a significant increase in BP of WT but not KO mice (DBP, WT: 89.2± 2.6 vs WT+Leptin 95.7±3.3; MAP, WT: 104 ±2.8 vs WT+Leptin: 110 ±2.7, n=5, p<0.05). We quantified sympathetic contribution to BP elevation by measuring BP response to glanglionic blockade (Hexamethonium). At baseline, KO mice exhibited a lower BP response than WT supporting a reduced neurogenic control of BP regulation in KO mice. Vascular contribution to high BP was investigated using wire myography in thoracic aorta. LepR deficiency impaired endothelial-dependent relaxation (EDR) to acetylcholine (n=7, p<0.05). L-NAME completely abolished EDR in KO and WT indicating that EC LepR deficiency reduced NO bioavailability. Recent evidence presents PFKFB3-mediated EC glycolysis as a new regulator of endothelial homeostasis. We found that aortic EC from KO exhibited increased PFKFB3 mRNA expression (p=0.065) and PFKFB3 inhibition restored EDR in KO. Remarkably, overexpression of PFKFB3 increased EC glycolysis
in vitro
and impaired EDR in WT aortic rings
ex vivo
. Collectively, our data suggest that impaired endothelial leptin receptor signaling induces a PFKFB3-dependent hyper-glycolytic phenotype resulting in NO deficiency and endothelial dysfunction that predisposes to higher BP regardless the reduced sympatho-activation which might prevent the increase in BP induced by exogenous leptin.
Abstract P277: β-arrestin-2-mediated Vasodilatation In Mouse Mesenteric Arteries
Authors: Pulgar Victor M , Toth Krisztian
Published: August 2021

As part of GPCRs-dependent signaling,
β
-arrestin-2 has been shown to stimulate eNOS activity and thus has the potential to modulate vascular function. We hypothesized that the absence of
β
-arrestin-2 would alter vascular dilatation and contraction in resistance arteries. We tested acetylcholine (ACh)-dependent relaxation and phenylephrine (PE)-dependent contraction in mouse mesenteric arteries isolated from 3-mo old male C57Bl6 (WT, n=5) and
β
-arrestin-2 KO (

βarr2
-/-

, n=5) mice. Segments were mounted in a Wire Myograph (DMT) for determination of isometric force; vessels studied included intact, without endothelium, or pre-incubated with L-NAME (10
-4
M). Dose-response curves were performed for ACh (10
-10
-10
-4.5
M) and PE (10
-10
-10
-4.5
M). Data were acquired using a PowerLab (ADInstruments) system. Maximal response to ACh (ACh
MAX
) was expressed as maximal relaxation after pre-constriction, maximal response to PE (PE
MAX
) as % of contraction to 75mM KCl (%K
MAX
), and sensitivity as pD
2
(-Log[EC
50
]). Data were analyzed using Prism (GraphPad). After pre-constriction (PE, 3x10
-6
M), arteries from

βarr2
-/-

mice presented similar ACh
MAX
(79±6
vs.
82±6, p>0.05) and lower sensitivity to ACh compared to WT (6.66±0.2
vs.
7.12±0.1, p<0.05). The sensitivity of the contraction to PE was increased in

βarr2
-/-

arteries (6.4±0.2
vs.
6.04±0.1, p<0.05), with no changes in PE
MAX
. Differences in vasodilation and contraction were abolished in arteries without endothelium and in arteries pre-incubated with L-NAME. We conclude that the absence of
β
-arrestin-2 induces a pro-contractile phenotype in an endothelium- and nitric oxide-dependent manner in mouse resistance arteries.
Abstract P278: Coronary Microvascular Dysfunction Is Associated With Augumented Autophagy-lysosomal Signaling In Mice With Hypercholesterolemic Diet
Authors: Wang Yun-Ting , Zhao Wei , Huckaby Ashton A , Li Xiang , Zhang Yang
Published: August 2021

Accumulating evidence indicates coronary microvascular dysfunction (CMD) contributes to myocardial ischemia with or without epicardial coronary atherosclerosis. However, it remains unknown which molecular pathway is associated with compromised coronary microvascular function preceding the development of myocardial ischemic injury. Recent studies suggest that autophagy-lysosomal signaling pathway is involved in the regulation of endothelial homeostasis under various metabolic stresses such as hypercholesterolemia. In this study, the early effects of hypercholesterolemia on the function and integrity of coronary microcirculation were studied in mice and the expressions of various molecular markers of autophagy-lysosome pathway were also determined in the coronary circulation. Mice were fed a hypercholesterolemic paigen diet (PD) for 8 weeks and coronary microvascular function was determined by measuring coronary flow reserve (CFR) under baseline and hyperemic conditions. The effects of PD on cardiac function or remodeling were also assessed by echocardiograph or immunohistochemistry studies. In PD-fed hypercholesterolemic mice, hyperemia-induced increase in CFR was significantly abrogated compared to that in normal chow diet-fed (ND) control mice (PD: 1.583±0.4193 vs. ND: 3.087±0.586) (n=7-8). The diet-induced hypercholesterolemia did not lead to cardiac dysfunction (EF%, PD: 59.870±7.549 vs ND: 64.040±9.088) or hypertrophic remodeling (LV mass (mg), PD: 92.240±14.410 vs ND: 96.030±25.07). PD increased mild cardiac inflammation and fibrosis but did not resulted in rarefaction in the myocardium. In small coronary arterial wall, PD induced endothelial inflammasome activation and inflammation, which was accompanied by upregulation of autophagy and lysosome signaling pathway. In conclusion, hypercholesterolemic diet induces CMD without alterations in cardiac function or remodeling. These alterations in coronary microvascular function represent the early effects of diet-induced hypercholesterolemia, which may be ameliorated with activation of autophagy and lysosome signaling pathways.
Abstract T2: Agreement Between Attended Home Blood Pressure Measurements And Ambulatory Blood Pressure Monitoring
Authors: Glenn Trevor , Eaton Cyd K , Psoter Kevin J , Eakin Michelle N , Pruette Cozumel S , RIEKERT KRISTIN , Brady Tammy M
Published: August 2021

Objectives:
Compare attended home blood pressure (BP) measurements (HBPM) with ambulatory BP monitor (ABPM) readings and examine if level of agreement between the measurement approaches differ overall and by subgroup.

Methods:
This was a secondary analysis of data from a 2-yr observational study of children 11-19 yrs (mean 15, SD=2.69) with chronic kidney disease. Participants had 3 standardized resting oscillometric home BPs taken by staff followed by 24-hr ABPM within 2 weeks of home BP. BP indices (measured BP/95%ile BP) were calculated for the mean triplicate attended HBPM and mean daytime ABPM readings. All paired HBPM and ABPM measurements taken during any of 5 study visits were compared using linear regression with robust standard errors. Generalized estimating equations-based logistic regression determined the sensitivity, specificity, negative, and positive predictive values with ABPM as the gold standard. Analyses were conducted for the group overall and by subgroup.

Results:
There were 103 participants who contributed 251 paired measurements. Indexed systolic BP did not differ between modalities (mean difference -0.002; 95% CI: -0.006, 0.003), and the difference in indexed diastolic BP was minimal (mean difference -0.033; 95% CI: -0.040, -0.025). The overall agreement between HBPM and ABPM in identifying abnormal BP was high (88.5%), and findings were consistent in each subgroup (Table).

Conclusions:
Attended HBPM may be a reasonable substitute for ABPM when monitoring BP. The greater accessibility and feasibility of attended HBPM has the potential to improve BP control among at-risk youth.
Abstract T6: Adipocyte Bardet-Biedl Syndrome 1 Gene Is Critical For Blood Pressure Regulation
Authors: Zhao Yuying , Guo Deng-Fu , Rahmouni Kamal
Published: August 2021

Bardet-Biedl syndrome (BBS) proteins have emerged as critical regulators of various physiological functions including blood pressure. The presence of hypertension in BBS patients is consistent with the finding that BBS knockout mice exhibit increased renal sympathetic nerve activity and arterial pressure. However, the role and contribution of
Bbs
genes in various tissues such as adipose tissue to blood pressure regulation remains unclear. To address this, we generated a novel conditional knockout mouse that lacks the
Bbs1
gene in adipocytes by breeding mice bearing floxed alleles of the
Bbs1
gene (Bbs1
flox/flox
) with mice expressing Cre specifically in adipocytes (Adiponectin
Cre
mice). Cre-mediated recombination was verified by loss of
Bbs1
expression in fat pads, but not in other tissues such as the kidneys, liver and skeletal muscle. Next, we assessed whether adipocyte
Bbs1
gene deletion affects body weight and glucose homeostasis. Interestingly, no body weight phenotype was observed in both male and female Adipo
Cre
/Bbs1
flox/flox
mice compared to their control littermates (males: 40.5±2.9 g vs 37.4±5.1 g; female: 27.7±4.0 g vs 27.5±3.6 g; 16 weeks old). However, insulin tolerance test uncovered impaired insulin-induced glucose clearance in both male and female Adipo
Cre
/Bbs1
flox/flox
mice. Subsequently, we measured blood pressure and found that Adipo
Cre
/Bbs1
flox/flox
mice have a remarkable increase in systolic blood pressure (130.4±11.2 mmHg) compared to the control littermates (119.1±4.5 mmHg,
p
<0.05). Finally, measurement of the expression of the renin-angiotensin system components revealed significantly elevated angiotensinogen mRNA (2.43±1.31 AU vs 1±0.72 AU,
p
<0.01) and angiotensin-converting enzyme mRNA (2.96±2.51 AU vs 1±0.95 AU,
p
<0.05) in peri-renal adipose tissue, but not in other fat pads such as brown and inguinal adipose tissues. Interestingly, gene expression of angiotensin receptor 1a was not significantly altered by
Bbs1
gene deletion. Taken together, our findings demonstrate that
Bbs1
gene in adipose tissue play an important role in the control of insulin sensitivity and blood pressure.
Parthian moγ and Middle Persian moγ/mow in Light of Earlier Eastern and Western Iranian Sources
Authors: Panaino Antonio
Published: August 2021

The present article analyses the historical importance assumed by Parthian and Middle Persian moγ/mow (and related words) in the framework of the religious and administrative language of Late Antiquity despite its seemingly absolute absence in the Avestan Sprachgut. Although moγ should be reasonably considered as a word of (prominent) Western Iranian derivation, i.e. from Median and Old Persian magu-, the progressive phonetic evolution toward a spelling, such as that of early Parthian and Middle Persian *moγ(u)- created a fitting resonance with a rare Avestan word (in its turn probably nonexistent in the older strata of the language, if not even a Western loanword itself), specifically moγu-°, which is attested only in the Y.Av. compound moγu.tb̰ iš-. The rising weight assumed by the priestly college of the Magi in secular activities already during the Achaemenian period promoted the preservation of this title also after the diffusion of the Avestan liturgy in Western Iran. This development also ensured that the designation of *moγ(u)- became extended to the whole family of the Zoroastrian priests following the Avestan tradition.
Iran’s Security Interests and Policies in the South Caucasus
Authors: Monshipouri Mahmood , Heiran-Nia Javad
Published: August 2021

The aim of this essay is to demonstrate that cooperation and competition between Russia, Iran, and Turkey requires ongoing regional political recalibrations regarding oil and gas pipelines and transportation routes. We argue that while much has changed in regional politics, Russian domination of the region remains intact, with Tehran and Ankara finding themselves in constant competition with each other while also balancing their interests versus those of Russia. We first review oil and gas pipelines administered by the Republic of Azerbaijan, the major beneficiary of the U.S. sanctions on Iran and the recent Nagorno-Karabakh war. We then explore a comparative analysis of the security interests and strategies of Iran, Russia, and Turkey toward the South Caucasus. Finally, we examine the way in which the second Nagorno-Karabakh war has posed new challenges to Iran’s interests and policies in the region.
Delivery of MiRNA-92a Inhibitor Using RP1-Linked Peptide Elicits Anti-Inflammatory Effects in an Acute Lung Injury Model
Authors: Zhuang Chuanyu , Piao Chunxian , Choi Myoungjee , Ha Junkyu , Lee Minhyung
Published: August 2021

Acute lung injury (ALI) is an inflammatory lung disease. miRNA-92a (miR92a) is induced in the lungs of ALI patients and mediates inflammatory reactions. In this study, a RP1-linked R3V6 (RP1R3V6) peptide was synthesized and evaluated as a carrier of anti-microRNA-92a oligonucleotide
(AMO92a) into the lungs of an ALI animal model. In addition to the carrier function, the RP1-linked peptide can have anti-inflammatory effects in the lungs, since RP1 is an antagonist of the receptors for advanced glycation end-products (RAGEs). In a gel retardation assay, the RP1R3V6 peptide
formed a spherical complex with AMO92a. In an in vitro delivery assay to L2 rat lung epithelial cells, RP1R3V6 had a lower AMO92a delivery efficiency than R3V6 and polyethyleneimine (PEI25k; 25 kDa). However, RP1R3V6 had an additional anti-inflammatory effect, reducing tumor necrosis
factor-α (TNF-α) in lipopolysaccharide-activatedmacrophage cells. With the combined effects of AMO92a and RP1, the RP1R3V6/AMO92a complex reduced the miR92a level more efficiently than did the R3V6/AMO92a and PEI25k/AMO92a complexes. The RP1R3V6/AMO92a complex was
administered into the lungs of ALI animals by intratracheal instillation. As a result, the expression of phosphatase and tensin homolog, a target of miR92a, was increased in the lungs. Furthermore, the RP1R3V6/AMO92a complex decreased the TNF-α and interleukin-1β (IL-1β)
levels more efficiently than did the PEI25k/AMO92a and R3V6/AMO92a complexes, decreasing the damage in the lungs. These results suggest that RP1R3V6 is a useful carrier of AMO92a and has anti-inflammatory effects in an ALI animal model.
Lipidated Methotrexate Microbubbles: A Promising Rheumatoid Arthritis Theranostic Medicine Manipulated via Ultrasonic Irradiation
Authors: Zhao Zhuofei , Lin Xiaona , Zhang Lulu , Liu Xia , Wang Qingwen , Shi Yu , Cui Guanghui , Cai Huali , Chen Yan , Li Yongbin , Hu Azhen , Zhang Zhuxia , Liu Jun , Xie Haiqin , Zheng Tingting , Liang Xiaolong , Shuai Xintao , Chen Yun , Sun Desheng
Published: August 2021

De novo designed lipidated methotrexate was synthesized and self-assembled into microbubbles for targeted rheumatoid arthritis theranostic treatment. Controlled lipidatedmethotrexate delivery was achieved by ultrasound-targetedmicrobubble destruction technique. Methotrexate was
dissociated inflammatory microenvironment of synovial cavity, owing to representive low pH and enriched leucocyte esterase. We first manipulated methotrexate controlled release with RAW 264.7 cell line in vitro and further verified with rheumatoid arthritis rabbits in vivo. Results
showed that lipidated methotrexate microbubbles precisely affected infection focus and significantly enhanced rheumatoid arthritis curative effect comparing with dissociative methotrexate. This study indicates that lipidated methotrexate microbubbles might be considered as a promising rheumatoid
arthritis theranostics medicine.
Rapid Detection of DNA Methylation with a Novel Real-Time Fluorescence Recombinase-Aided Amplification Assay
Authors: He Ziyu , Tong Zengrui , Tan Boyu , He Xuliang , Zhang Tao , Guo Yuan , Jin Lian , He Nongyue , Li Song , Chen Zhu
Published: August 2021

Researchers have conducted in-depth research on DNA methylation mechanism, which is related to various diseases such as deficiency of imprinted gene and occurrence of tumors. This study provides a novel rapid quantitative detection assay and real-time fluorescence recombinase-aided amplification assay (RAA) for DNA methylation. Firstly, specific sequence of methylation genes was chosen and primers and fluorogenic probe for RAA experiment were designed and synthesized. Lastly, these amplification products were proven by sequencing and analysis. Results showed that the amplification efficiency and template concentration of RAA had linear dependent (R2> 95%) when the concentration range was 4.64×108copies/μL˜4.64×104copies/μL. The test assay can also detect positive samples when the template concentration is below 4.64×104copies/μL. Remarkably, the entire experiment process only takes 15–20 minutes, so it is beneficial for rapid bedside simple screening of some special DNA methylation sites, such as detection of resistance genes. In a word, this method has very great potential for diseases with DNA methylation in clinical settings, especially if methylation analysis needs to be done quickly and easily.
Titanium Cage in Comparison with Nano-Hydroxyapatite Bone Graft Substitutes in Cervical Reconstruction
Authors: Zhang Tao , Guo Ying , Zhang Dapeng , Zhao Runze , Hu Naiwu
Published: August 2021

Cervical spondylosis is a disease that occurs with age and affects the discs and joints in the cervical spine located in the neck. The hydroxyapatite (HA) bone graft substitutes can used as a potential bone-forming agent, however, the efficacy of using HA is challenging in cervical
reconstruction. In this regard, nano-based HA was used in this study to explore its sagittal parameters and clinical potency in relative to titanium (TiO2) cage in patient with cervical spondylosis. 50 patients suffering from cervical spondylosis were divided in two groups and were
grafted with either TiO2 cage or nano-HA. The sagittal parameters, including cervical spine lateral radiographs (C0−2Coob and C2−7Coob) were taken pre- and post-operation (3-month, one-year and two-year). The clinical potency was also done based
on the JOA scores. Angle analysis indicated that the C0−2Cobb and C2−7Cobb angles were significantly changed after the operation in both TiO2 cage and nano-HA groups, whereas no significant changes was determined in nano-HA relative to TiO2
cage condition. Also, it was shown that JOA scores were significantly higher after the operation than pre-operation, indicating a potential cervical reconstruction after surgery in both groups which slightly were higher for nano-HA groups.