bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P218: Ume (
Prunus Mume
) Attenuates Er Stress And Inflammation In Rat Vascular Smooth Muscle Cells
Authors: Kuroda Ryohei , Okuno Keisuke , Cicalese Stephanie M , Ueda Yuriko , Okuno Yoshiharu , Kono Ryohei , Utsunomiya Hirotoshi , Marumoto Shinsuke , Eguchi Satoru
Published: August 2021

Ume (
Prunus mume
), is a traditional and familiar food in Asian countries, and is most often pickled or used to make wine and juice concentrate. Recently, Ume is attracting attention as a health food. We previously reported that Ume attenuates the activation of EGFR-ERK cascade and ROS production induced by angiotensin II (Ang II) in vascular smooth muscle cells (VSMC) of thoracic aorta. However, how Ume works on vascular inflammation and ER stress still remains unknown. AngII may enhance inflamm-aging via ER stress contributing to arterial stiffness and hypertension.We used primary culture of rat VSMC for experiment. We obtained 5 fractions (methanol, hexane, dichloromethane, ethyl acetate, and water) extracted from Ume juice. VSMC were pretreated with each fraction for 30 minutes followed by Ang II or tumor necrosis factor (TNF)-α stimulation. Stimulation period was 24 and 48 hours for Ang II, 6 hours for TNF-α. As the preliminary screening, we tested the cell response and solubility of each fraction (n=1~2). Pretreatment with 500μg/mL hexane fraction (PMF-H) showed the tendency to decrease VCAM1 expression under Ang II condition. PMF-H was completely soluble in the media at the concentration of 150 μg/mL. Based on this result, we focused on 150μg/mL PMF-H. DMSO was used as vehicle. In Western blot analysis, PMF-H attenuates ATF6 expression under both AII (p=0.0351, n=5) and TNF-α (p=0.0199, n=5) condition compared to vehicle. VCAM1 expression was significantly enhanced by Ang II stimulation (p=0.0471, n=5), and attenuated by PMF-H. PMF-H also reduced VCAM1 expression under TNF-α stimulation compared to vehicle (p=0.0377, n=5). GRP78 and Crystallin AB did not show any significant change. ATF6 and VCAM1 are key modulators of ER stress and vascular inflammation that may lead to cellular aging. Our results indicate PMF-H attenuates ATF6 and VCAM1 induced by Ang II or TNF-α stimulation. Thus, identification of the active ingredient in hexane fraction of Ume is desired as it may constitute a good candidate for a supplement or drug for anti-inflamm-aging therapy.
Abstract P220: Disparity In Hospital Admissions And Length Of Stay Based On Income Status For Emergency Department Hypertensive Crisis Visits: A Cross-sectional Study Using The Nationwide Emergency Department Sample
Authors: Srivastava Shreya , Vemulapalli Bhargav , Okoh Alexis K , Kassotis John
Published: August 2021

Introduction:
Racial, gender and lower socioeconomic status have been shown to negatively impact the delivery of care. How this impacts the management of hypertensive crisis (HC) remains unclear.

Objective:
Identify disparities on admission frequency and length of stay (LOS) among those presenting with HC, as a function of household income.

Methods:
This is a cross-sectional analysis of 2016 ED visits and supplemental Inpatient data from the Nationwide Emergency Department Sample. Median household income quartiles were established. A multivariable logistic regression model was used to estimate odds of admission in each income quartile. A multivariable linear regression model was used to predict LOS.

Results:
After applying sample weighting, the total number of ED visits was 33,728 with 25442, 6906, and 1380 visits for hypertensive urgency (HU), emergency (HE) and unspecified crisis, respectively. There were 13191, 8889, 6401, 5247 visits in the (1
st
) lowest, 2
nd
, 3
rd
and 4
th
(highest) income quartiles, respectively. The median age was 61 and 58 years for HU and HE, respectively. The most common comorbidity was chronic kidney disease. Individuals with the highest income, had a lower odds of admission compared to the lowest quartile [Adjusted Odds Ratio: 0.41, 95% CI: 0.22,0.74] (
Figure 1a
). There was a significant linear association between income quartile and LOS across all HC and HE [beta coefficient: 0.411, 0.407 p value = 0.015, 0.019] (
Figure 1b
).

Conclusions:
In this study, patients with lower income were more likely to be admitted, while those with higher income exhibited a longer LOS. Clinicians must be made aware these disparities to ensure the equitable delivery of care.
Abstract P221: Effect Of Instructional Videos To Enhance Healthcare Provider Knowledge For Hypertension (htn) Management In Tanzania
Authors: Edward Anbrasi , Kagaruki Gibson , Manase Frank , Matsushita Kuni , Appel Lawrence J
Published: August 2021

Introduction:
The growing burden of HTN is a major public health problem in Tanzania, with the estimated prevalence of 28%. The healthcare system faces major capacity constraints, including inadequate provider competencies; <60% of providers reported fair knowledge of HTN, and only 42% had access to HTN guidelines. Knowledge of HTN management standards and guidelines is prerequisite to improving HTN management.

Objective:
Determine the effect of instructional training videos in improving healthcare provider knowledge for HTN management.

Methods:
Eighteen primary healthcare facilities in the Dar es Salaam region were selected using stratified random sampling. In each facility, providers, who screened and managed patients for HTN, completed pre-test knowledge assessments. Subsequently, they were invited to an instructional training, using four, 4-5minute videos developed by investigators at Johns Hopkins. Post-test was conducted 5 months after the introduction of the videos.

Results:
Thirty-three clinicians participated in the pre-post study and consented to attending the video training workshop. Knowledge competencies showed significant improvement in most but not all aspects of HTN management (Table). For example, adequate knowledge about HTN increased from 78.8% to 100% (p<0.01), and knowledge of patient preparation prior to Blood Pressure (BP) screening increased from 69.7% to 93.9% ( p<0.05).

Conclusion:
In the context of a resource-constrained setting, low-cost training videos have the potential to improve healthcare provider knowledge for HTN management.
Abstract P222: Impact Of Adverse Childhood Experiences On Ambulatory Blood Pressure Profiles In Adolescence
Authors: Kellum Cailin E , Perry Paige , Kemp Keri , Johnson Alan K , Pollock David M , Mrug Sylvie , Pollock Jennifer , Seifert Michael , Feig Daniel
Published: August 2021

Adverse childhood experiences (ACEs) include exposure to abuse (verbal and physical), neglect, and household dysfunction during childhood. ACEs have long-lasting health impacts including increased risk for cardiovascular disease (CVD) and hypertension in adulthood. However, it is not clear how ACE exposure impacts CVD risk earlier in the life course, particularly in adolescence. To address this gap in knowledge, in this study we hypothesized that ACE exposure is associated with abnormal ambulatory blood pressure (ABP) profiles in adolescents, with an increased incidence of ambulatory hypertension phenotypes that have normal casual clinic BP [e.g., masked hypertension (MH) or blunted nocturnal dipping (BND)]. We utilized 24-h ambulatory BP monitoring (ABPM; Spacelabs) and casual clinic BP to construct a profile of adolescents with and without ACEs. Abnormal ABP profiles included the following categories: ambulatory hypertension (AH, elevated ABP and casual clinic BP ≥95
th
percentile for age, sex, and height), white-coat hypertension (WCH, elevated casual clinic BP with normal ABP), MH (normal casual clinic BP with elevated ABP), or BND (drop in ABP < 10% during sleep). This study included 78 male and female adolescents (median age=16) recruited from Children’s of Alabama Pediatric Clinics. Exclusion criteria included known CVD and antihypertensive medication. Participants recorded wake and sleep times in a diary. Based on the ACE questionnaire, 51 (65%) of adolescents experienced at least 1 ACE. The prevalence of abnormal ABP profiles was similar between the group with ACE exposures vs. the group without ACE exposures (34% vs. 36%;
P
=0.87). In participants with ACE exposure (n=51), 9% had AH, 6% had MH, 19% had WCH, 43.1% had systolic BND and 22% had diastolic BND. In participants without ACEs (n=27), 4% had AH, 4% had MH, 29% had WCH, 37% had systolic BND and 15% had diastolic BND. Further analysis with covariates are necessary. These results suggest that adolescents with ACEs have similar prevalence of abnormal ABP overall, but higher prevalence of individual ABP phenotypes such as AH, MH, and BND compared to adolescents without ACEs.
Abstract P223: Vitamin D Supplementation Improves Measures Of Arterial Stiffness And Vascular Function In Adolescents Suffering From Orthostatic Intolerance
Authors: Shaltout Hossam A
Published: August 2021

Background:
In previous work we identified a group of adolescents with orthostatic intolerance (OI) presenting as postural orthostatic tachycardia syndrome or syncope with impairment in autonomic and vascular response upon head up tilt. Low vitamin D level correlated with the severity of symptoms. In this pilot study we hypothesized that vitamin D supplementation will improve the vascular function in these adolescents.

Methods:
A cohort of twenty Adolescents (mean age= 16.2 years, 4 males) who are vitamin D deficient had a non-invasive measurements of brachial BP, aortic BP, augmentation index at heart rate of 75 beats per minute (AIx75), and carotid-femoral pulse wave velocity (cf-PWV) using the SphygmoCor XCEL System at baseline and after two months of vitamin D supplementation (2000-5000 IU daily based on baseline level). Both right and left side ankle pressure and ankle brachial index (ABI) were assessed using COLIN VP-1000 vascular profiling system. Impedance Cardiography (ICG) was used to estimate total arterial compliance (TAC) and thoracic fluid content (TFC) in supine position at rest and after hand grip challenge.

Results:
As shown in the table. Compared to baseline, vitamin D supplementation increased vitamin D level, increased TAC and TFC at baseline and under stress challenge. It also tended to reduced AIx75 and cf-PWV and reduced both right and left ankle pressure and ABI

Conclusions:
Vitamin D supplementation improved different measures of vascular function in adolescent. These data provides evidence of potential therapeutic benefit of vitamin D supplementation for patients suffering from orthostatic intolerance who are vitamin D deficient.
Abstract P226: Transcriptome Signature Of Placentas From Diabetic Pregnancies Is Confounded By Fetal Sex.
Authors: Kedziora Sarah M , Obermayer Benedikt , Sugulle Meryam , Herse Florian , Kräker Kristin , Haase Nadine , Langmia Immaculate , Müller Dominik N , Staff Anne , Beule Dieter , Dechend Ralf
Published: August 2021

Hypertension-associated obstetric diseases like diabetes mellitus (DM) or preeclampsia occur in 25-35% of pregnancies and are related to placental dysfunction. The placenta has a unique structure and function to guarantee healthy pregnancy, fetal development, and outcome. Thus, the diabetic environment featuring inflammation, metabolic alterations, and hypoxia, is challenging with increased stress for mother and foetus. Delivery-related complications and adverse post-pregnancy outcomes, for example the increased risk for preeclampsia, are related to placental pathology in the different types of DM. Therefore, understanding disturbed placental development during pathologic pregnancy is important. We hypothesize a placental transcriptome signature present in pathologic pregnancy. We aimed to broaden our understanding of placental adaptations to various forms of diabetic pregnancy. Human placenta samples from healthy controls (n = 29), women with gestational DM (GDM, n = 12), DM type 1 (DM1, n = 17), DM type 2 (DM2, n = 3) and GDM, DM1 or DM2 superimposed by preeclampsia (n = 16) were included. We quantified gene expression by Illumina TruSeq stranded total RNA Sequencing and analyzed data by means of principal component analysis (PCA), differential expression and gene set enrichment. The samples were mainly split by fetal sex in the PCA rather than diabetic subgroups. Genes associated with the principal components were primary located on the male sex chromosome (
Xist, Uty, Usp9y, Ddx3y
) and only weakly enriched in functionally relevant gene sets. The fetal sex was identified as single main explanatory variable when the principal components were associated with clinical patient data. Placentas complicated by DM2 identified 93 and 27 significant differential expressed genes (DEG; adj. p-value < 0.05) in comparison to healthy controls and GDM, respectively. Comparison of DM1 to DM2 or GDM showed four and two significant genes. In this cohort possible effects of GDM and preeclampsia were not observed.We conclude that fetal sex is dominating placental gene expression and confounding transcriptome signatures resulting from diabetes, in settings of well controlled diabetic disease.
Abstract P227: Gq Signaling In The Placental Syncytiotrophoblast Layer During Preeclampsia
Authors: Opichka Megan , Livergood Mary Christine , Sigmund Curt D , McIntosh Jennifer , Grobe Justin L
Published: August 2021

Hormones implicated in preeclampsia (PE) such as angiotensin, endothelin, and vasopressin signal via receptors coupled to the Gq cascade, and Regulator of G protein Signaling-2 (RGS2) buffers this signaling. We have published that RGS2 expression is decreased in human PE placenta, and reducing RGS2 in placenta causes development of key features of PE in mice. New in situ hybridization data indicate that in both humans and mice, RGS2 is abundant among many cell types in the placenta, including the syncytiotrophoblast (STB) layer. In addition, RGS2 expression in the human STB layer is reduced during PE. As this layer is strongly implicated in PE, these data lead us to hypothesize a critical Gq-buffering role for RGS2 in STB cells to prevent PE. To explore the effect of excess Gq signaling within the STB layer, we utilized a Cre-Lox approach to cause expression of the Gq-coupled hM3Dq DREADD throughout the fetoplacental unit (dam: hM3Dq+, sire: Actb-Cre+) or only within the STB layer (dam: hM3Dq+, sire: Gcm1-Cre+), and then activated the hM3Dq receptor via clozapine N-oxide (CNO, 0.5 to 2 mg/kg) injection in mid-gestation (GD12.5-14.5) before tissue collection at GD14.5. Gαq activation throughout the fetoplacental unit (Actb-Cre model) severely restricted fetoplacental growth compared to saline-injected controls (n=2 vs 3; placenta: 0.027±0.006 vs 0.115±0.021 g; p<0.05, and fetus: 0.048±0.007 vs 0.268±0.010 g; p<0.05). Similarly, placentas expressing hM3Dq only in STB cells (Gcm1-Cre model) had reduced placental (n=3 0.116±0.022 vs 0.201±0.036 g; p=0.05) and possibly fetal (n=3 0.1112±0.036 vs 0.247±0.028 g; p=0.06) masses after CNO. Vascularization (assessed by CD31 immunostain) was disproportionately reduced in the labyrinth layer of the Actb-Cre model after CNO (n=2 vs 3; 20.189±3.382 vs 35.762±1.976 % area; p<0.05), despite no relative change in layer (ie, decidua/junctional zone/labyrinth) thicknesses. Preliminary results indicate similar findings in the Gcm1-Cre model (n=1 17 vs 25 % area). These data highlight the pathological consequence of excess Gq signaling in the STB layer. Ongoing studies are aimed at characterizing maternal phenotypes in these models and the consequence of STB-specific deletion of RGS2 upon sensitivity to Gq stimulators.
Abstract P233: Microvascular Fingerprint Of Multi-Organ Imaging - New Insights In Preeclampsia Research
Authors: Kraeker Kristin , Napieczynska Hanna , Kedziora Sarah , Haase Nadine , Müller Dominik N. , Heuser Arnd , Dechend Ralf
Published: August 2021

To establish the first protocol for high-resolution three-dimensional imaging of multiple organs in rodents, we performed microvasculature casting with Microfil silicone rubber substance in non-living rats and combined the method with advanced image analysis protocols after micro-CT scanning. This newly established method allows changes in individual organs to be visualized and examined in extraordinary detail as part of a multi-organ approach. Previously published protocols only address visualization of single organs or are insufficiently resolved with respect to vessel diameter analysis. With optimized staining protocols, samples can additionally be examined for possible structural changes such as inflammatory processes or fibrotic formation. Preeclampsia is a condition during pregnancy that leads to up to a 4-fold increased risk of maternal cardiovascular disease later in life, although symptoms usually resolve after delivery. In former preeclamptic rats, a reduction in microvascular density in the heart and brain with a simultaneous increase in retinal vessels has been described, even weeks after the disappearance of major features such as hypertension and albuminuria. The technique described will enhance studies of the vasculature in many other animal models and provide striking insights into vascular physiology necessary to drive translation.
Abstract P241: Salt Directly Alters Molecular Trafficking Of Uromodulin In Renal Thick Ascending Limb Cells
Authors: Boder Philipp , Mary Sheon , Mullen William , Graham Delyth , Mark Patrick B , Leiper James , Delles Christian
Published: August 2021

Uromodulin (UMOD) is the most abundant renal protein secreted into urine by the thick ascending epithelial (TAL) cells of the loop of Henle. Genetic studies have demonstrated an association between UMOD risk variants and hypertension. We have previously demonstrated that salt induces a reduction in urinary UMOD excretion in normotensive Wistar-Kyoto (WKY) rats after 3 weeks, which was exacerbated in a chronic hypertensive rat model with increased accumulation of UMOD in the endoplasmic reticulum (ER). We further showed that these effects occur at the protein trafficking level. Our objective was to understand whether UMOD trafficking is influenced by acute salt exposure. Further, we aimed to dissect the molecular pathways through an untargeted proteomics approach. To study acute effects, WKYs were subjected to 1% NaCl on an intermittent basis over 3 weeks in an on-off-on pattern (control n=4, salt n=6). In an
ex vivo
experiment, TAL tubules isolated from WKY were incubated with salt (+300mOsm) (control/salt n=3). Protein extractions were labelled with isobaric tandem mass tags and analysed by mass spectrometry. Acute salt-loading significantly lowered UMOD excretion after one week (9-fold, p<0.05), which returned to control levels when halted, and decreased again when resumed a week later (4-fold, p<0.05). Together, this confirmed that salt directly influences UMOD trafficking even after a brief exposure. Proteomic analyses of
ex vivo
tubule incubated with salt revealed 45 differentially expressed proteins (p<0.05), of which 15 were upregulated (>1.2 fold) in the salt group. Gene ontology analysis results showed that these differentially expressed proteins were enriched in the mitochondria/electron transport chain (NDUFS6, FDX1, NDUFV2, TXNRD2), oxidation-reduction processes (PP1R1A, SUOX, CYB5R1), and protein transport (EXOC6, MACF1, GLG1, SORT1, BNIP1). In summary we demonstrate that UMOD excretion responds to changes in exposure to salt
in vivo
and
in vitro
and that altered UMOD trafficking may be due to changes in mitochondrial function and oxidative stress. The exact role of the identified proteins in UMOD trafficking warrants further study.
Abstract P242: The Role Of Kappa Opioid Receptors In The Development Of Hypertension And Kidney Injury In Sprague-Dawley Rats
Authors: Golosova Daria , Zietara Adrian , Bohovyk Ruslan , Levchenko Vladislav , Staruschenko Alexander
Published: August 2021

The extensive use of opioid-based pain management strongly correlates with poor cardiovascular and cardiorenal outcomes. Our recent studies suggest that treatment with kappa opioid receptor (KOR) agonist BRL 52537 leads to the progression of chronic kidney disease (CKD) and aggravation of salt-sensitive hypertension. We hypothesize that stimulation of KORs leads to blood pressure elevation, albuminuria, and kidney damage in healthy Sprague-Dawley (SD) rats. To characterize the effect of the KOR agonist BRL 52537 on the development of blood pressure and kidney function
in vivo
, SD rats were treated with a daily
i.v.
bolus infusion of BRL 52537 or a corresponding vehicle. To test the contribution of KOR stimulation on calcium homeostasis in podocytes, BRL 52537 was used on freshly isolated glomeruli from SD rats. Single-channel analysis was applied to assess the effect of KORs stimulation on TRPC6 channel activity in the human immortalized podocytes. Chronic treatment with BRL 52537 leads to increased mean arterial pressure (88±1 vs 101±4 mmHg, vehicle vs treated, p<0.05), podocyte basal calcium (90±12 vs 216±16 a.u., vehicle vs treated, p<0.05), and GFB impairment in SD rats which is reflected by a transient increase in albumin excretion (Alb/cre ratio 0.35±0.1 vs 0.72±0.2, vehicle vs treated, p<0.05). Cumulative probability distribution analysis of the glomerular injury score revealed a rightward shift toward a high glomerular injury score in the group treated with BRL 52537 (p<0.05). Angiotensin II level was higher in a BRL-treated group (156±17 vs 232±59 pmol, vehicle vs treated, p=0.065); however, it did not reach a statistical difference. Acute application of BRL 52537 resulted in sustained calcium response (0.23±0.01 a.u., Fluo4/FuraRed, maximum calcium response) in freshly isolated glomeruli from SD rats. Furthermore, patch-clamp experiments in human immortalized podocytes (cell-attached configuration) revealed that BRL 52537 activated TRPC6 channels. Taken together, these data support the hypothesis that administration of opioids in SD rats leads to activation of the KOR/TRPC6 pathway, which in turn led to glomerular filtration barrier impairment, increased glomerular damage, and blood pressure elevation.
Abstract P245: Activation Of Protease-activated Receptors 1 Leads To Structural Changes In Immortalized Cultured Human Podocytes.
Authors: Bohovyk Ruslan , Khedr Sherif , Klemens Christine A , Levchenko Vladislav , Isaeva Olena , Palygin Oleg , Staruschenko Alexander
Published: August 2021

Previous studies have suggested that activation of the protease-activated receptors (PAR) exacerbates the development of diabetic nephropathy. PARs are transmembrane proteins activated by extracellular serine proteases, such as thrombin and trypsin, which are frequently elevated in disease states. We hypothesize that serine protease activation of the PAR1 signaling cascade upregulates calcium channel activity and promotes excessive intracellular Ca
2+
([Ca
2+
]
i
), leading to podocyte foot retraction, apoptosis, and glomerular filtration barrier (GFB) damage. Immunofluorescent labeling revealed the co-localization of PAR1 and podocyte marker nephrin in freshly isolated human glomeruli. To explore the functional role of PAR1 mediated signaling, we used an immortalized cultured human podocyte (hPod) cell line. Live confocal imaging revealed rapid elevation of [Ca
2+
]
i
in response to a PAR1 specific agonist (TFLLR-NH
2
). Moreover, preincubation with a PAR1 antagonist (RWJ 56100) completely blocked this effect (1616±48 vs. 206±135 a.u., for TFLLR vs. TFLLR+RJW, n<100 cells, p<0.05). Furthermore, we tested potential downstream signaling proteins in hPod cells after PAR1 activation using Western blot analysis. Application of TFLLR-NH
2
led to a decrease in PAR1 expression due to internalization and lysosomal degradation, accompanied by an increase in TRPC6 expression and time-dependent changes in p-ERK1/2 and PLC-γ1 cascades. We further used scanning ion-conductance microscopy (SICM) to detect morphological changes in podocytes, which measures real-time surface topography in hPod cells. This imaging revealed normal protrusion of lamellipodium under control conditions. In contrast, PAR1 activation led to retraction of the lamellipodium, and a significant decrease in surface area (+51±46 vs. -15±8 μm
2
, 30 min after application of vehicle or TFLLR, respectively; n=5, p<0.05). Our studies suggested that PAR1 is involved in GPCR-induced calcium influx in podocytes. Furthermore, these data demonstrate that PAR1-mediated signaling is involved in podocyte structural alterations and may increase [Ca
2+
]
i
, potentially leading to loss of GFB integrity and podocyte apoptosis.
Abstract P254: Sex Differences In Angiotensin Ii-induced Hypertension In Mice With Proximal Tubule-specific Deletion Of Mitochondrial Protein Sirtuin 3 In The Kidney
Authors: Leite Ana Paula O , Li Xiao C , Hassan Rumana , Chen Jian-xiong X , Sato Akemi , Dugas Courtney M , Sato Ryosuke , Zhuo Jia L
Published: August 2021

The development of Angiotensin II (Ang II)-induced hypertension is associated with mitochondrial dysfunction and kidney injury. Sirtuin 3 (SIRT3), a key mitochondrial protein, plays an important role in maintaining mitochondrial homeostasis. However, it remains unknown whether deletion of SIRT3 in the proximal tubules will alter the pressor and renal responses to Ang II in sex-different manners. In the present study, adult male, and female wild-type (WT) and mutant mice with proximal tubule-specific knockout of SIRT3, PT-
Sirt3
-/-
, were infused with or without a slow pressor dose of Ang II via an osmotic minipump (0.5 mg/kg/day, i.p.), supplemented with a 2% NaCI diet or losartan, 20 mg/kg/day, for 2 weeks. Systolic (SBP), diastolic (DBP), and mean arterial blood (MAP) pressure were determined using the tail-cuff method, whereas 24 hr. urinary sodium and potassium excretion were determined using a metabolic cage. Serum and urine creatinine were measured using colorimetric assays, whereas glomerular and tubulointerstitial injury was evaluated by Masson’s Trichrome staining. Basal SBP levels were lower in PT-
Sirt3
-/-
than in WT mice (SBP-WT: 112 ± 2 vs. SBP-PT-
Sirt3
-/-
: 93 ± 2 mmHg,
P
<0,01). The magnitude of Ang II-induced hypertension was similar between WT and PT-
Sirt3
-/-
mice with or without losartan treatment. Serum creatinine levels and urinary creatinine excretion were higher in PT-
Sirt3
-/-
mice than in WT mice (
P
<0.05), but without significant sex differences in response to Ang II infusion or losartan treatment. Differences were found only in female WT and PT-
Sirt3
-/-
mice with lower 24 hr. urine and urinary creatinine excretion in response to Ang II infusion or losartan. Losartan significantly increased 24 hr. urinary potassium and chloride excretion in Ang II-infused male and female PT-
Sirt3
-/-
mice (
P
<0.01). Finally, Ang II-infused PT-
Sirt3
-/-
mice showed significant renal cortical tubulointerstitial fibrotic responses (
P
<0.05), but not glomerular fibrotic responses. We conclude that basal blood pressure is lower in male and female PT-
Sirt3
-/-
mice and that Ang II induces similar hypertensive and renal fibrotic responses in male and female PT-
Sirt3
-/-
mice without significant sex differences.
Abstract P257: Refractory Hypertension Partially Improved With Anxiolytics
Authors: Al-Qudah Bara , Ahmed Ashraf E , Alamin Mohammed , Aboolmaaly Cheikh , Shariff Mohamed
Published: August 2021

Background:
Resistant hypertension is a BP that is not controlled by three or more anti-hypertensive medications of different classes, including diuretics. Refractory hypertension is an evolving term reflecting a BP that is not controlled with five agents or more of different anti-hypertensive classes, including diuretics. Patient with refractory HTn has to have secondary hypertension workup.

Case:
A 45-year-old lady with a known past medical history of obesity, HTN, and DM-II. She was admitted as she had generalized fatiguability and vague chest pain. The ECG, and the TTE showed hypertensive heart disease. She had normal cardiac enzymes. Initial BP was recorded to be 185/127 mmHg. Consequently, she was commenced on anti-HTN medications. Her BP was difficult to control with the maximum doses of six anti-HTH medications (amlodipine, valsartan, hydralazine, indapamide, labetalol, spironolactone)(Fig 1).

Decision-making:
All the secondary causes of HTN were excluded; normal renin-aldosterone ratio, normal US doppler for the renal arteries, normal sleep study, normal CT chest and abdomen looking for masses, normal dotatate scan, and negative pheochromocytoma work-up. Because she had social stress, she was commenced on anxiolytics (alprazolam). Subsequently, the BP reading improved by around 10-20 SBP; however, her average reading continued to be around 160/90 mmHg.

Conclusion:
Refractory hypertension is a management dilemma, especially when the causes of secondary hypertension are excluded. Benzodiazepines can be a reasonable option in certain situations where the refractory hypertension is believed to be stress-induced.
Abstract P259: Cardiovascular Pharmacotherapy And Covid-19
Authors: Lip Stefanie , McCallum Linsay , du Toit Clea , Kilmartin Jason , Murray Eleanor , Reetoo Salil , Knox Laura , Rostron Maggie , Guzik Tomasz J , Delles Christian , Dominiczak Anna F , Touyz Rhian M , Padmanabhan Sandosh
Published: August 2021

COVID-19 illness is associated with cardiovascular vulnerabilities at all stages pre-infection, acute phase and subsequent chronic phase. The major cardiometabolic drivers identified to have epidemiological and mechanistic associations with COVID-19 are abnormal adiposity, dysglycaemia, dyslipidaemia, and hypertension. Recent findings demonstrate that cardiovascular risk reduction interventions may have short-term benefits on COVID-19 risk and outcomes. Our aim was to investigate the effect of cardiovascular pharmacotherapy on SARS-CoV-2 infection. Demographic data, cardiovascular disease and other comorbidities, prescription data, discharge outcomes, RT-PCR tests, and investigations were obtained through record linkage for all patients admitted between 01/04/2020 and 31/05/2020 to NHS Greater Glasgow & Clyde hospitals. The relationship between cardiovascular pharmacotherapy (antihypertensives and statins) and SARS-CoV-2 infection was analysed using multivariable logistic regression models. From 218,472 patient records available in Glasgow SafeHaven. 3,610 patients had confirmed COVID-19 based on clinical criteria or a positive RT-PCR test) and 18,050 propensity matched controls were selected. The average age was 62 years, 55% (1,985 of 3,610) females, 43% (1,552 of 3,610) resided in the most deprived areas (SIMD:1). There were 46% (1,660 of 3,610) on antihypertensive therapy and 35% (1,264 of 3,610) were on statin therapy. The risk of SARS-CoV-2 infection was greater in those with cancer, pulmonary or renal disease in the multivariable logistic regression model. Those on antihypertensive or statin therapy had an 11% (O.R.[95% CI]: 0.89 [0.82-0.97], p=0.007) and 9% (0.91 [0.83-0.99], p=0.03) lower risk of SARS-CoV-2 infection, respectively. The protective effect of statins and antihypertensive drugs were apparent only in those on both drug therapies (0.82 [0.74-0.92], p=0.001). Prior treatment with statins and antihypertensive drugs are associated with a lower risk of SARS-CoV-2 infection. The observed beneficial effect of statins and antihypertensives therapy warrants further investigation to address critical gaps in knowledge in prevention, treatment and long term follow up of patients with COVID-19.
Abstract P266: Hypertensive Response In Recently Covid-19 Vaccinated Patients
Authors: SANIDAS ELIAS , PAPADOPOULOS DIMITRIOS , MANTZOURANI MARINA , VELLIOU MARIA , ANASTASIOU THEOCHARIS , FOTSALI ANASTASIA , BARBETSEAS JOHN
Published: August 2021

Background:
A non-immediate hypertensive response after COVID-19 vaccination has been reported. Mild to moderate elevated arterial blood pressure (BP) levels have been documented a few days after a single or two-doses vaccine. This study sought to investigate this observation as a potential side effect in patients with known hypertension and healthy controls.

Methods:
Overall 60 vaccinated patients between the age of 50 to 70 years old were studied. They were randomly assigned to one of the approved and available vaccines (Pfizer, Astra Zeneca or Moderna) twice within a month. Half of them were hypertensives under medical treatment and half of them were not.All participants volunteered for standard daily home blood pressure measurements (HBPM) and had also ambulatory blood pressure measurements (ABPM) between the 1
st
and the 30
th
day after the second dose of COVID-19 vaccine.

Results:
All patients, hypertensives or not, had at some point a recorded and substantial hypertensive response for both systolic (SBP) and diastolic (DBP) blood pressure after the second dose of the vaccine. Hypertensives were older and with higher body mass index (BMI). Some of the hypertensive patients received additional medication whereas some of the non-hypertensive patients started life modification changes and systematic BP measurements for a possible diagnosis of hypertension.

Conclusions:
Vaccination for COVID 19 seems to be related with a short period of hypertensive response. This phenomenon was partial and mostly observed in older overweight hypertensives.
Abstract P267: Arbidol, A Targeted Covid-19 Therapy, Inhibited Contractile Responses To Phenylephrine In Mesenteric Arteries And Aortas Of Hypertensive Rats
Authors: Arishe Olufunke O , Wilczynski Stephanie , Thamasett Anna , Webb Clinton R
Published: August 2021

Background:
Hypertension is the most prevalent comorbidity of COVID-19 infection. This infection is caused by the severe acute respiratory syndrome coronavirus SARS-CoV-2. SARS-CoV-2 is a single-stranded RNA and could activate the endosomal Toll-like receptor 7 (TLR7), indicating a vital role of the receptor in its therapeutic target. Studies have reported that the lysosomotropic drug chloroquine has antihypertensive effects through its action on inhibiting the endosomal TLRs. Arbidol is a potential broad-spectrum antiviral used in the treatment of many viral infections including influenza. Arbidol prevents viral infection by interacting with aromatic acids; arbidol also binds to lipid membranes, altering the configuration of the cytoplasm or endosome. Wang
et al.,
in 2020 reported that arbidol effectively inhibited the coronavirus SARS-CoV-2 in vitro by impeding viral attachment and release from the endolysosomes.

Hypothesis:
We hypothesized that arbidol would improve vascular reactivity in hypertension.

Methods:
We studied the effects of arbidol on the contractility of mesenteric resistance arteries (MRA) and aortas of male Wistar rats (20 weeks old, n=4) and spontaneously hypertensive rats (SHR) (20 weeks old, n=4) on a myograph by incubating the arterial rings with 10μM arbidol (Sigma, USA.) or vehicle (DMSO) for 30 mins. Statistical analysis was performed using nonlinear regression, and one- and two-way ANOVAs.

Results:
Arbidol (10 μM) impaired the contractile responses of MRA and aorta from normotensive and hypertensive rats to phenylephrine (PE). MRA Wistar: (pEC50 6.11 ± 0.15 vs 3.42 ± 0.38), SHR: (5.89 ± 0.19 vs 3.56 ± 0.19) and aorta Wistar: (pEC
50
7.49 ± 0.19 vs 5.97 ± 0.14), SHR (pEC50 7.14 ± 0.31 vs 4.41 ± 0.15). The arbidol-induced impaired contraction to PE was greater in the SHR aorta compared with the Wistar aorta. After washing out the drug, the arterial rings contracted to 120mM KCl at the same magnitude as the initial contraction to 120mM KCl.

Conclusions:
Our data demonstrate that arbidol impairs vascular contractile responses to α-adrenergic stimulation, with a greater response in the hypertensive rats. This indicates antihypertensive effects potentially through the regulation of TLR signaling.
Abstract P268: The Impact Of Covid-19 On A Large Pragmatic Clinical Trial Embedded In Primary Care
Authors: Hau Cynthia , Leatherman Sarah , Klint Alison , Glassman Peter , Taylor Addison , CUSHMAN WILLIAM , Ishani Areef
Published: August 2021

Introduction:
The COVID-19 pandemic has had a significant impact on traditional clinical trial operations, but it is unclear whether the impact persists in pragmatic trials with a centralized study design. The Diuretic Comparison Project (DCP) is a Point-of Care pragmatic trial that operates through a usual care system to compare chlorthalidone and hydrochlorothiazide at preventing major cardiovascular (CV) events and non-cancer death. We assessed the impact of the COVID-19 pandemic on the centralized recruitment, patient follow-up, data collection, and outcome ascertainment performed in the DCP.

Methods:
We assessed operations in two 8-month periods: Pre-COVID-19 (Jul 2019 - Feb 2020) and Mid-COVID-19 (Jul 2020 - Feb 2021). Enrollment, study medication adherence, blood pressure (BP) and electrolyte follow-up rates, VA records of CV events, all-cause hospitalization, and death rates were compared.

Results:
Providers agreed to participate at a lower rate, but more patients were contacted and randomized during mid-COVID-19. While BP evaluations decreased, the rates of electrolyte, major CV, and medication prescription records were comparable to the pre-COVID-19 period (
Table 1
).

Conclusions:
The DCP was able to recruit and maintain critical data collection at the pre-COVID-19 levels. There were some decreases in BP evaluations, likely due to fewer in-person visits. All-cause VA hospitalizations also decreased, despite rises in COVID-related hospitalizations and death. While the impact on outcome and safety rates awaits complete data from Medicare, the DCP has demonstrated a promising centralized design that can support pragmatic trial operations during a pandemic.
Abstract P279: Glycocalyx Restoration Reduces Arterial Stiffness In Diabetic Female Mice
Authors: Manrique Acevedo Camila , Ramirez-Perez Francisco , Soares Rogerio N , Jurrissen Thomas J , Ghiarone Thaysa , Morales-Quinones Mariana , Cabral-Amador Francisco J , Vink Hans , Foote Christopher A , Martinez-Lemus Luis A , Padilla Jaume
Published: August 2021

Arterial stiffening, a characteristic feature of type 2 diabetes, is an important contributor to the development and progression of cardiovascular disease (CVD). Thus, a better understanding of the precipitating factors underlying arterial stiffening is vital to identify newer targets and strategies to reduce CVD burden, particularly in diabetic women who exhibit heightened arterial stiffening and more severe CVD. Degradation of the endothelial glycocalyx in diabetes is thought to contribute to endothelial dysfunction and CVD development. However, whether glycocalyx degradation is also an important determinant of arterial stiffening remains unknown. Herein, we hypothesize that restoration of the glycocalyx with dietary supplementation of glycocalyx precursors (DSGP, including glucosamine sulfate, fucoidan, superoxide dismutase, and high molecular weight hyaluronan; Endocalyx
TM
) improves endothelial function and lessens arterial stiffness in diabetic female mice. To test this hypothesis, we used 12-week old db/db female mice that were treated with DSGP (100 mg/kg/day) or vehicle (
i.e.
, peanut butter) for four weeks, and an age-matched db/+ cohort as reference control. After euthanasia, we assessed
ex vivo
aortic stiffness and glycocalyx length via atomic force microscopy. Using pressure myography, we also determined
ex vivo
mesenteric artery endothelial function and stiffness by measuring flow-mediated dilation and the passive mechanical properties of the arterial wall, respectively. Consistent with our hypothesis, vehicle-treated db/db mice exhibited degradation of the endothelial glycocalyx, impaired endothelium-dependent vasodilation, and increased arterial stiffness when compared with control db/+ females. Moreover, treatment with DSGP was effective at restoring the endothelial glycocalyx in db/db mice. Notably, this restoration of the glycocalyx was accompanied with improvements in endothelial function and reductions in arterial stiffness. Collectively, these findings support the notion that the endothelial glycocalyx should be considered as a putative therapeutic target to reverse arterial stiffening in diabetic females.
Abstract P280: CCL5 Causes Vascular Injury Via CCR5 And Nadph Oxidase 1- Derived Ros
Authors: Singh Shubhnita , Bruder Nascimento Ariane , Bargaje Anita , Bruder Nascimento Thiago
Published: August 2021

Chemokine (C-Cmotif) ligand 5 (CCL5) and its receptor CCR5 belong to the family of chemokines and are expressed and active in the vasculature. NADPH oxidases (Noxs) are the major source of reactive oxygen species (ROS) in vascular cells, but whether the activation of these oxidases is CCL5/CCR5 sensitive and whether such interaction participates in the genesis of vascular disease is not fully known. We investigated whether CCL5/CCR5 leads to vascular injury by activating Nox1. Carotid ligation model (CL, for 2-weeks) was used to induce pathological vascular growth in 10-weeks old (C57BL6/J) mice. Rat aortic smooth muscle cells (RASMC) were treated with recombinant CCL5 (100ng/mL) to study the molecular mechanisms. CL induces neointima formation, which was associated with increase in IL1β, TNFα, CCR3, CCR5 (3-fold increase), CCL5, and Nox1 gene expression. No difference was observed for Nox2 and 4. Treatment with CCR5 blocker (maraviroc, 25mg/Kg/day i.p) partially inhibited CL-induced vascular injury (media/intima ratio, CL: 1.2 ± 0.2 vs CL + maraviroc: 0.7 ± 0.2) and Nox1 expression (Fold changes: CL: 2.1 ± 0.4 vs CL + maraviroc: 1.2 ± 0.4). In RASMC, CCL5 induced Nox1 expression, which was blunted by pre-treating cells with maraviroc (10uM). Also, it increases p47phox content in membrane fraction (index of Nox activation), and elevated ROS production, analyzed by L012. CCL5 also induced cell migration, measured by transwell assay (number of cells per spot, control: 21.3 ± 3.1 vs CCL5: 31.1 ± 2.4), proliferation, analyzed by Edu+ cells (% of cells per spot, control: 10.6 ± 4.3 vs CCL5: 22.8 ± 5.1), and inflammation (studied by IL1β and TNFα levels). Lastly, CCL5 elevated NF-κB translocation into the nucleus, indicating NF-κB activation. Strikingly, inhibition of Nox1 (GKT771, 10uM), blocked CCL5-induced vascular migration, proliferation, and inflammation, as well as NF-κB activation. We propose that CCL5 activates Nox1 in the vasculature leading to local injury characterized by vascular inflammation and cellular migration and proliferation, perhaps by activating NF-κB signaling. Herein, we place CCR5 signaling as possible therapeutic target to reduce the cardiovascular risk in inflammatory diseases associated with dysregulation of CCL5 and/or CCR5
Abstract P281: Vascular Remodeling And Impaired Vascular Smooth Muscle Cell Plasticity In Age And Sex-dependent Hypertension
Authors: Amraei Razie , Nist Kayla , Moreira Jesse , Wainford Richard D
Published: August 2021

Aim:
Hypertension (HTN) and aging are associated with the development of vascular dysfunction. We speculated that vascular smooth muscle cell plasticity and vascular remodeling play major roles in age and sex-dependent HTN.

Methods:
Male and female Sprague-Dawley (SD) rats aged 3 and 16-months-old (N=6/group) were housed under standard conditions. Blood pressure was measured via femoral artery cannulation and sympathetic tone to the vasculature was estimated by ganglion blockade via Hexamethonium (30mg/Kg IV). PBS-perfused abdominal aorta and renal arteries were collected and immunoblotting was performed following protein extraction.

Results:
Male SD rats, but not females, develop HTN and increased sympathetic tone with age. Aged hypertensive male rats, but not aged normotensive females, exhibit reduced p-Erk1/2 and p-eNos levels in both abdominal aorta and renal arteries. α-Smooth Muscle Actin significantly increased in aged male abdominal aorta. Elevated c-Src was observed in aged female abdominal aorta and p-c-Src was reduced in aged male abdominal aorta. Caveolin-1 changed oppositely in young and aged abdominal aorta and renal arteries of two sexes.

Conclusions:
Our data suggest that artery-specific changes of key signaling molecules contribute to impaired vascular smooth muscle plasticity and vascular dysfunction in aged hypertensive male but not in aged normotensive female rats.
Abstract P284: The Association Of Oral Hypofunction And Increased Arterial Stiffness
Authors: Matsumoto Chisa , Tomiyama Hirofumi , Matsuura Mari , Nakai Takayuki , Chikazu Daichi , Yamashina Akira , Kawai Takashi
Published: August 2021

Background:
Frailty is associated with higher risk of cardiovascular disease (CVD) and mortality. Recently, oral hypofunction, a disease in which the oral function is complexly reduced not only by ageing but also by a variety of factors, is regarded as a major risk factor for frailty, as it develops malnutrition and sarcopenia. However, no studies have evaluated the association of oral hypofunction and arterial stiffness, a marker for CVD.

Hypothesis:
We hypothesized that subjects with oral hypofunction have increased arterial stiffness compared to those without oral hypofunction.

Methods:
Japanese subjects above 50 years old who underwent annual health checkup were enrolled in this cross-sectional study. Subjects with history of CVD and dementia were excluded. Arterial stiffness was evaluated by brachial-ancle pulse wave velocity (baPWV). Oral hypofunction was evaluated based on the guidance by the Japanese Association for Dental Science. Seven oral factors, oral hygiene, oral moisture, occlusal force, tongue-lip motor function, tongue pressure, masticatory function, and swallowing function were assessed. Oral hypofunction was diagnosed if more than 3 factors showed deterioration. The association of oral hypofunction and baPWV was evaluated by multivariate linear regression analysis adjusted for conventional CVD risk factors. We also performed subgroup analysis stratified by age. (<60, ≧60 years).

Results:
Among 148 subjects (mean age: 59±7 years), 34 subjects (23%) had oral hypofunction. BaPWV in subjects with oral hypofunction was significantly higher than subjects without oral hypofunction (mean baPWV: 1539±312 v.s. 1416±260cm/sec, p=0.02). However, after adjustment for CVD risk factors, oral hypofunction did not significantly associate with baPWV (β=46±49, p=0.35). On the other hand, in subgroup analysis, oral hypofunction was significantly associated with increased baPWV among subjects in subjects younger than 60 years old even after adjustment of CVD risk factors (β=135±67, p=0.046), but this association was not observed in subjects over 60s.

Conclusion:
Oral hypofunction was associated with increased arterial stiffness, especially in subjects younger than 60s. Further research on oral hypofunction and CVD is warranted.