bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P089: Health Facility Readiness For Hypertension Management In Ghana, West Africa
Authors: Byiringiro Samuel , Commodore-Mensah Yvonne , Hinneh Thomas , SARFO FRED S , Dennison Himmelfarb Cheryl R
Published: September 2022

Background:
Hypertension continues to be a growing health burden in Sub-Saharan Africa (SSA). The health systems in SSA are ill-equipped to manage patients with hypertension. The current study explored hypertension service readiness at the health facilities in Kumasi, Ghana.

Methods:
We purposively recruited 11 (out of over 200) health facilities that provide hypertension services in Kumasi, Ghana. At each facility, we administered the Hypertension Service Availability and Readiness tool. The main outcome of interest was hypertension service readiness regarding the availability of hypertension guidelines, provider and their training, equipment, diagnostics, and anti-hypertensive medications. We used descriptive analysis and reported counts (%) and median (Interquartile Range).

Results:
Five facilities (46%) were of medium size with 50-100 bed-capacity, four (36%) were government-owned, five (46%) treated patients with hypertension in a hypertension clinic while the rest managed them in the general outpatient department (Table). At least one provider had received cardiovascular disease training at 7 (64%) facilities. Hypertension-related patient education materials and treatment guidelines were available in 6 (54%) and 8 (72%) facilities, respectively. Also, 7 (64%) had all of the first-line anti-hypertensive medications (as stipulated in Ghana Standard Treatment Guidelines) available.

Conclusion:
We identified gaps in provider training, patient education, and first-line medication availability related to hypertension in Ghana. This study calls for policy changes to improve hypertension management and prevention of cardiovascular diseases.
Abstract P091: Role Of Gata3 In Renin Cell Identity
Authors: Neyra Jesus S , Medrano Silvia , Sequeira Lopez Maria , Gomez Roberto A
Published: September 2022

Renin cells act as precursors for other cell types in the kidney vasculature and show high plasticity in postnatal life in response to challenges to homeostasis. Our scRNA-seq studies revealed that the dual-zinc finger transcription factor (TF) Gata3, important for cell lineage commitment and differentiation in several tissues, is expressed in mouse renin cells under normal conditions and homeostatic threat. In addition, we found an enhancer associated with Gata3 in renin cells leading us to hypothesize that GATA3 is important for renin cell identity. We studied 60 and 76 day-old mice carrying a conditional deletion of Gata3 in renin cells: Gata3fl/fl;Ren1dCre/+ (Gata3 cKO), and control Gata3fl/fl;Ren1+/+ counterparts. Gata3 cKO mice exhibited increased BUN (43±4.6mg/dl) vs. control mice (BUN: 21±2mg/dl), suggestive of hypovolemia and/or compromised renal function. By immunostaining, renin expressing cells appeared very thin compared to the normal plump shape in control mice. Renin signal was abnormally localized to the Bowman’s capsule in some glomeruli, and there was an atypical strong Acta2 signal in the mesangium and Bowman’s capsule in Gata3 cKO mice. Distal tubules showed dilated morphology without visible intraluminal casts whereas proximal tubules appeared normal. Gata3 cKO recruited mice exhibited marked reductions in Ren1+ cells compared to controls by an 87% reduction of Ren1 mRNA levels by RT-qPCR (1.68 ± 0.11 vs 12.98 ± 0.98, p=0.0006). H&E, PAS, and Mason’s Trichrome reactions demonstrated increased glomerular fusion, absent cubical epithelial cells in Bowman’s capsule, and intraglomerular aneurysms.

In summary, we found abnormal expression and localization of Ren1 and Acta2 and abnormal morphology of kidneys in Gata3 cKO mice. Our results suggest a role of Gata3 in the identity of cells of the renin lineage. Future studies will investigate the role of Gata3 during development and in older animals.
Abstract P097: Effect Of Maternal Fecal Microbiome And Adverse Pregnancy Outcomes In The Obese BPH/5 Female Mice
Authors: Beckers Kalie F , Schulz Christopher , Childers Gary , Sones Jennifer L
Published: September 2022

Obesity is a worldwide pandemic and of women childbearing age in the US, two-thirds are considered overweight/obese. Maternal microbial dysbiosis associated with obesity may play a role in adverse pregnancy outcomes. To better understand the relationship of microbiome and adverse pregnancy outcomes, we utilized the obese hypertensive BPH/5 mouse model of preeclampsia (PE). We hypothesized that the maternal fecal microbiome during pregnancy will differ in the BPH/5 mice compared to lean normotensive C57 controls as well as non-pregnant (NP) BPH/5 females. We have shown that adult BPH/5 female mice have an adverse cardiometabolic phenotype: hypertension and obesity that is exaggerated by pregnancy and phenocopies PE. To test our hypothesis, fecal samples were collected from NP and pregnant (P) BPH/5 and C57 control females in late gestation and Illumina sequencing of 16S v4 rRNA amplicon libraries were used to compare microbiomes. Microbial community composition of BPH/5 compared to C57 P mice were significantly different (variance of 33.59% in PCo1 and Pco2 11.48%, n=9-17, p=0.001, PERMANOVA with Bray-Curtis dissimilarity of 16S Amplicon Sequence Variant’s relative abundance). Alpha diversity was increased in BPH/5 compared to C57 P mice (n=9-17, p=0.005). There was a difference in beta diversity within the communities in BPH5 P mice compared to BPH5 NP mice, suggesting that the fecal microbiome was altered with pregnancy (n=9-17, p=0.006), but no difference was seen in the alpha diversity meaning that the richness and evenness were similar (n=9-17, p=0.115). The most abundant phyla are represented by 49.39% Firmicutes, 38.54% Bacteroidetes, and 7.48% Epsilonbacteraeota. The most abundant genera including 22.59%
Alistipes,
19.57%
Lachnospiraceae
genus unknown, and 9.85%
Helicobacter
within the BPH5 samples.
Muribaculum spp
(p=1.58e-06) and
Lachnospiraceae_NK4A136_unclassified
(p=2.46e-03) showing significant decreases with pregnancy. In conclusion, obese BPH5 dams have altered fecal microbial communities associated with obesity and adverse pregnancy outcomes. Further investigations are necessary to further understand the importance of these microbial community changes and how they play a role in maternal hypertension and offspring.
Abstract P100: Combination Treatment Of Resveratrol And Pravastatin Decreases NF-κB Expression In HUVEC Model Of Preeclampsia
Authors: Thamasett Anna R , Arishe Olufunke O , Webb R Clinton , Lane-Cordova Abbi D
Published: September 2022

Introduction:
Preeclampsia (PE) is a hypertensive disorder of pregnancy characterized by new-onset hypertension with new-onset proteinuria or other signs/symptoms of preeclampsia after 20 weeks’ gestation. It is associated with an increase in anti-angiogenic, oxidative, and pro-inflammatory factors including soluble fms-like tyrosine kinase-1 (sFLT-1). While a previous study established cumulative beneficial effects of resveratrol and pravastatin on cardiovascular disease, no study has been conducted regarding this combination therapy on PE.

Hypothesis:
The combination of resveratrol and pravastatin is more effective in reversing endothelial damage than the individual drugs alone.

Methods:
Human umbilical vein endothelial cells (HUVECs) were incubated with sFLT-1 (25ng/ml) for 24 hours. HUVECs were then treated with either 20μM resveratrol, 20μM pravastatin, 20μM resveratrol + 20μM pravastatin, or left untreated. Cells were lysed and western blotting was performed using primary antibodies against: eNOS, phospho-eNOS and NF-κB. Immunoreactive bands were visualized and quantified using Image Studio Lite Ver 5.2.

Results:
Pre-treatment with sFLT-1 increased expression of NF-κB compared to untreated controls (Mean±SEM: 53.57±24.98 vs 27.78±15.85 p=0.0346), while combination treatment of sFLT-1 pre-treated cells reduced expression of NF-κB (Mean: 53.57±24.98 vs 38.99±26.81 p=0.0186). eNOS expression was not changed by pre-treatment with sFLT-1, but expression was improved by pravastatin only in control cells (Mean: 3.084±0.6266 vs 4.737±1.247 p=0.0463). Phospho-eNOS expression was improved by pravastatin treatment in both control cells (Mean: 1.309±0.4633 vs 2.467±0.8298 p=0.0213) and in sFLT-1 pre-treated cells (Mean 1.235±0.3765 vs 2.716±0.9470 p=0.0347). Finally, phospho-eNOS expression was improved by combination treatment in control cells (Mean: 1.309±0.4633 vs 1.825±0.5793 p=0.0136).

Conclusions:
Our results show that treatment with resveratrol and pravastatin was more effective at decreasing inflammation than individual treatments, therefore suggesting that the combination therapy could be a helpful treatment for PE pathophysiology.
Abstract P102: A History Of Preeclampsia Is Associated With Accelerated Aging And Multimorbidity Later In Life
Authors: Suvakov Sonja , Vaughan Lisa , Parashuram Santosh , Mielke Michelle , Bielinski Suzette J , Chamberlain Alanna M , Kantarci Kejal , Garovic Vesna D
Published: September 2022

Background:
Preeclampsia (PE) is a pregnancy-specific hypertensive disorder which affects up to 7% of all pregnancies. It is associated with increased risk for cardiovascular disease later in life. We have recently reported increased senescence and accelerated aging in women with PE. We hypothesize that senescence burden remains after PE. The aim of this study was to compare senescence profiles, and history of individual chronic conditions and multimorbidity, a marker of accelerated aging, in women with and without a history of PE.

Methods:
Forty women with histories of PE and 40 with histories of normotensive pregnancy confirmed by manual chart review were age- and parity-matched. None of the women had a history of CVD at the time of recruitment. Markers of senescence (p16, p21 and Klotho) and metabolic syndrome (leptin, adiponectin) were measured and compared between women with and without histories of preeclampsia using conditional logistic regression. The rates of chronic conditions and multimorbidity between the groups were also compared.

Results:
Urinary Klotho/creatinine levels were significantly lower (median [IQR]: 117 [87.4, 224] vs. 205 [101, 359], respectively, p=0.042), while leptin/adiponectin ratio was significantly higher (median [IQR]: 0.25 [0.13, 0.45] vs. 0.05 [0.02, 0.12], p=0.027) among women with histories of PE compared to controls. There were no significant differences observed in plasma p21 or p16 concentrations. Women with histories of PE experienced a higher prevalence of two or more co-morbidities compared to women with normotensive pregnancies [67.5% (27/40) vs. 22.5% (9/40), p<0.05]. Results were similar after removing hypertension as a comorbidity [45% (18/40) vs. 20.0% (8/40), p<0.05].

Conclusion:
Women with a history of PE had altered markers of senescence, metabolic syndrome and multimorbidity decades after PE.
Abstract P104: Neutralization Of Marinobufagenin Demonstrates Efficacy In Vitro And In Vivo In Models Of Preeclampsia
Authors: Pantho Ahmed F , Afroze Syeda H , Wages John M , Yu Bo , Vora Niraj , Larrick James W , Beeram Madhava R , Zawieja David C , Uddin Mohammad N
Published: September 2022

Objective:
Marinobufagenin (MBG), a cardiotonic steroid, is distinctly elevated in preeclampsia (preE) and may directly contribute to pathogenesis, possibly through deleterious signaling in cytotrophoblast (CTB) cells. In this study, we evaluate the effects of anti-MBG human monoclonal antibodies on cellular signaling in CTB cells and in a rat model of preE.

Methods:
CTB cell proliferation in response to MBG with and without anti-MBG was measured using a Cell Titer assay. Pro-angiogenic factors (VEGF and PlGF) and anti-angiogenic factors (sFlt-1 and sEng) in response to MBG with and without antibody were measured. Finally, we evaluated the lead anti-MBG antibody in comparison with the parent murine antibody in a previously described rat model of preE which recapitulates the hypertension, proteinuria, and fetal growth effects of human preE.

Results:
CTB cells exposed to ≥ 1 nM MBG showed decreased (
p
< 0.05) proliferation, decreased secretion of VEGF and PIGF, and increased secretion of sFlt-1 and sEng. Pretreatment with anti-MBG significantly (
p
< 0.05) attenuated the MBG-induced modulation of cell proliferation and expression of VEGF, PIGF, sFlt-1, and sEng. In the rat model, anti-MBG treatment normalized blood pressure, reduced proteinuria, and eliminated fetal effects.

Conclusions:
MBG has the potential to be a causative agent for preE as it causes dysfunction in CTB cells due to the anti-angiogenic milieu. Our study suggests that anti-MBG antibodies can bind to MBG, neutralizing it, and preventing downstream signaling
in vitro.
In a saline-induced rat model of preE, treatment with anti-MBG antibody was effective at normalizing blood pressure, kidney function, and fetal birth weights. These data suggest that a human monoclonal antibody with high specificity and affinity for MBG has potential as a therapeutic for preE.
Abstract P113:
SOX6
IS NOT A RELEVANT TRANSCRIPTION FACTOR FOR RENIN TRANSCRIPTIONAL REGULATION BY JUXTAGLOMERUALR CELLS
Authors: Martini Alexandre G , Sequeira Lopez Maria L , Gomez Roberto A
Published: September 2022

The SRY (sex determining region Y)-box 6,
Sox6
, encodes a member of trancription factors (TFs) crucial for organogenesis. It has been suggested that
Sox6
is essential for the
Renin
transcriptional regulation. However, we identified
Sox6
expression only in tubular cells. We aim to investigate whether
Sox6
contributes to Renin gene expression by the Juxtaglomerular Cells in normal and under stress. Therefore, we generated Ren1
d
-Cre;Sox6
-/-
mice, where cells expressing
Renin
lacks the
Sox6
gene. We randomized the mice into four groups (n= 6/group): Wild-type (WT) and mutants (
Sox6
cKO) exposed to either regular salt diet or low salt diet and captopril (0.5 mg/mL in water
ad libitum
for 8 days). The later condition decreases blood pressure and increases
Renin
transcription and exocytosis. We measured the blood pressure by direct carotid artery catheterization. There were no differences between WT and cKO in mean blood, systolic or diastolic pressures. ELISA assay measured the plasmatic Renin content, without significance in any group (WT: 126775 pg/mL ± 59035; cKO: 200008 pg/mL ± 68556; n=3).
In sytu hybridization
(
ISH
) for
Sox6
has never been visible in the kidney despite all optimizations.
Renin
ISH
had usual distribution in the Juxtaglomerular Apparatus (JGA; n=6). In the captopril treated groups,
Renin
intensity was higher and no differences were noted. Immunohistochemistry for
Renin
also reavealed no differences between WT and cKO. Yet, the renin+ cells extended along the afferent arterioles in the captopril treated groups. We calculated the Juxtaglomerular Index, which is the total number of renin+ JGA divided by all glomeruli. WT showed 156 renin+ JGA of 441 glomeruli (n=3), while the cKO exhibited 173 of 449 glomeruli (n=3). In the captopril groups, WT had 302 of 452 glomeruli (n=3), while the ckO exhibited a subtle decrease, 176 of 312 glomeruli (n=4), but without significance. Quantitative RT-PCR from the kidneys showed an increase in the relative R
enin
gene expression to
s14
in cKO group (WT: 1.2 ± 0.8; cKO: 2.1 ± 2.2; n=3), while
Sox6
relative gene expression to
s14
decreased (WT: 1.02 ± 0.27; cKO: 0.35 ± 0.2; n=3). We conclude that
Sox6
is not an important TF for
Renin
transcriptional regulation by JuxtaGlomerular Cells in neither normal nor stressed conditions.
Abstract P116: Specific Chromatin Configurations Across The Genome Determine The Renin Cell Identity
Authors: Xue Bingjie , Medrano Silvia , Smith Jaosn , Martini Alexandre G , Sheffield Nathan , Sequeira-Lopez Maria Luisa , Gomez Roberto A
Published: September 2022

Renin cell descendants, such as smooth muscle cells, regain the endocrine renin phenotype to overcome homeostatic threats. The mechanism involved and the determinants of the renin cell identity are not well understood. We hypothesize that renin cell identity is governed by specific chromatin domain configurations genome wide. We compared open chromatin regions between renin- and non-renin-expressing cells. Non-renin control ATAC-seq data were mainly from ENCODE, containing 178 samples of 26 cell types. The differential analyses with DESeq2 between three renin groups (renin-expressing tumoral cell lines, native and recruited renin cells) and non-renin control group revealed that the chromatin configurations are different between the tumoral cell line and the primary kidney cells (native and recruited) in 1) number of differential regions (22,321 increased accessibility regions in the tumoral cell lines and 3,964 in the primary kidney renin cells); 2) enriched GO terms, and 3) distribution pattern over the genome. Within a small proportion of overlapped differential accessibility regions, all three groups have accessibility regions associated with known renin-specific genes: Ren1 and Akr1b7. The motif enrichment analyses with the MEME suite on the increased accessibility regions show different sets of enriched transcription factor (TF) motifs between the tumoral cell line and primary cells. The top enriched motifs in the tumoral cell line are the bZip family, including Atf3 (q-value: 2.56e-2147), Fos (q-value: 1.01e-2406), and JunB (q-value: 8.73e-2432). The top enriched motifs in the primary cells are the MEF2 family, including MEF2a (q-value: 2.18e-91), MEF2c (q-value: 1.28e-80), and MEF2d (q-value: 1.28e-80), consistent with unpublished work in our lab. In addition, we found four topmost enriched motifs (MEIS1, NR4A2, FOXJ3, and NR4A1, with q-values of 8.24e-10, 9.78e-9, 3.96e-7, and 2.67e-6) co-enriched in regions associated with Ren1 and Akr1b7. This indicates that a set of TFs regulates the co-expressing of Ren1 and Akr1b7. The chromatin domains and TF we described are likely to control renin cell identity and may serve as therapeutic targets for kidney/vascular diseases and hypertension.
Abstract P118: Metabolic And Hemodynamic Disorders In Obese Patients With True And Pseudo-resistant Hypertension
Authors: Shalimova Anna
Published: September 2022

Objective:
to conduct a comparative assessment between pseudo-resistant and true resistant hypertension (RH) in obesity.

Methods:
The study included 302 patients with uncontrolled hypertension and obesity. Initial treatment efficacy was assessed 3 months after dual therapy was administered. Those patients who did not reach target blood pressure (BP) in dual therapy were transferred to triple therapy. Among patients who received triple therapy, 69 people did not reach target BP (they received the fourth drug spironolactone). All patients were additionally examined 6 months after the initiation of antihypertensive therapy.

Results:
In the absence of a difference in BP levels, patients who subsequently become resistant, already at the stage of inclusion, significantly more often had circadian rhythm (CR) violations with a predominance of the non-dipper CR. Despite the target BP achievement after 6 months, in the majority (91.42%) of non-RH patients, the CR normalized, while in RH a normal CR was only in 60.87%. At the stage of enrolling patients into the study and 3 months after the start of therapy there was no significant difference in BP levels between true and pseudo-RH, then after 6 months of therapy, patients with true RH had significantly higher office SBP levels (p<0.01) and 24 h average SBP according to ABPM data (p<0.05) compared with pseudo-RH. Already at the stage of inclusion in the study, the normal CR in true RH was significantly less common than in pseudo-RH, and during the prescribed therapy, a similar trend persisted: after 6 months of therapy, 71.05% of pseudo-RH had a normal CR, while with true RH - only about half of the patients. Obese patients with true RH had also significantly lower BMI and LDL-cholesterol (p<0.05) as well as higher levels of aldosterone and SBP (p<0.05) compared with pseudo-RH.

Conclusions:
Even when target BP levels in antihypertensive therapy are achieved, obese patients with RH are characterized by more pronounced CR disturbances and higher levels of office and out-of-office BP, compared with non-RH. Compared with pseudo-RH, the presence of true RH in obesity is associated with higher SBP and aldosteron levels, as well as lower BMI and LDL-cholesterol.
Abstract P119: Severe Paroxysmal Hypertension In Patient With Gastric Neuroendocrine Tumor Detected By PET Dotatate Imaging
Authors: Ayzenbart Vira , Wasilewski Melissa , Schumaecker Matthew
Published: September 2022

Neuroendocrine tumors (NET) are rare neoplasms originating from neuroendocrine cells in the endocrine, gastrointestinal, or bronchopulmonary systems. When symptomatic, patients may present with paroxysmal HTN. A 51-year-old woman with CAD and prior CABG presented to the hospital with sudden onset headache and BP in the 200s/90s. Her physical exam was unremarkable. ECG showed new T wave inversions in leads V1 and V2 and high-sensitivity troponin was elevated at 1827 ng/L. CT angiography of her head and neck was normal. Her HTN improved with IV labetalol and pain control. We admitted her for BP control and secondary HTN evaluation. Initial lab findings were unremarkable (see table). A renal artery ultrasound showed no renal artery stenosis. CT of her chest, abdomen, and pelvis showed no evidence of NET. She continued to have paroxysms of severe HTN, initially controlled with IV medications, then transitioned to an oral four-drug regimen prior to discharge. Fractionated plasma metanephrines and catecholamines, and 24-hour urine fractional metanephrines were obtained (results pending at time of discharge).Follow up testing showed elevated urinary metanephrines and Chromogranin levels (see table), suggestive of NET. A Ga-68 PET-DOTATE scan showed a focus of increased activity in the anterior stomach. Endoscopy revealed a gastric body lesion, identified as grade 2, well-differentiated NET on biopsy. The patient underwent laparoscopic wedge gastrectomy with clear margins. Her headaches resolved, and BP improved. This case demonstrates the utility of Ga-68 PET DOTATE imaging in determining the presence of NET, not detected by CT images, as the reason for severe, paroxysmal HTN.
Abstract P123: Angiotensin Ii Type 1a Receptor Expressed Insmooth Muscle Cells Is Required Forangiotensin Ii-induced Vascular Remodeling But Not For Cardiac Hypertrophy
Authors: Okuno Keisuke , Torimoto Keiichi , Cicalese Stephanie , Sparks Matthew A , Eguchi Satoru
Published: September 2022

Angiotensin II Type 1a Receptor (AT1a) has been shown to play a critical role in cardiovascular diseases. We have demonstrated that ADAM17 mediates cardiovascular remodeling induced by angiotensin II (AngII) by using an SM22α Cre reporter, which expresses in smooth muscle cells (SMC) and cardiomyocytes. It is conceivable that AT1 signaling in SMC specifically initiates cardiovascular remodeling, such as hypertrophy and fibrosis. Deficiency of SMC AT1a receptors via endogenous SM22α promoter knock-in (KISMKO) results in diminished hypertension and cardiac hypertrophy induced by AngII. However, we have limited understanding whether SMC AT1a receptors affect vascular fibrosis and cardiac hypertrophy. Thus, this study was designed to elucidate the roles of the SMC AT1a receptor in cardiovascular remodeling using two distinct SMC AT1a receptor deficient mice. One line is the KISMKO, and another line is Tamoxifen inducible SMMHC-Cre AT1aflox/flox mouse (SMMHCKO) silencing in SMC but not cardiac myocytes. 8 to 12-week-old male mice were infused with AngII (1 μg/kg/min) for 2 weeks. Cardiac hypertrophy was assessed by heart-to-body weight ratios and found to be reduced in KISMKO mice compared with control mice (6.04 mg/g versus 4.89 mg/g, p=0.032), whereas ratios were not altered in SMMHCKO mice (5.31 mg/g versus 5.47 mg/g, p=0.941). Histological assessment demonstrated that perivascular fibrosis in coronary arteries was increased in control mice and attenuated in both KISMKO and SMMHCKO mice (1.27 vs. 0.22, p<0.0001 and 0.70 vs. 0.16, p<0.0001, respectively.). Medial thickness of descending aorta was increased in control mice and these phenotypic changes were attenuated in both KISMKO and SMMHCKO mice (97 vs. 56 μm, p<0.0001 and 98 vs. 74 μm, p<0.0001, respectively.). In conclusion, SMC AT1a receptors could mediate AngII-induced vascular remodeling including perivascular fibrosis but not cardiac hypertrophy. Cardiac myocyte AT1a appears critical for cardiac hypertrophy. Our data together with a past manuscript demonstrate a protective effect of SMC AT1a silencing in overall vascular remodeling. However, the exact relationship between hypertension and vascular remodeling remains to be elucidated.
Abstract P125: Long-term Exposure Of Angiotensin II Enhanced Vascular Contraction Via Activation Of Protein Kinase C Beta
Authors: Morikita Nayu , Mukohda Masashi , Nakamura Sho , Naganishi Sayaka , Mizuno Risuke , Ozaki Hiroshi
Published: September 2022

Protein kinase C (PKC) β is linked to lymphoma and diabetes complications including diabetic retinopathy, nephropathy and neuropathy. Also, recent studies revealed that PKCβ activation also accelerates atherosclerotic plaque formation in diabetes. These findings strongly indicate that PKCβ is a potential target for the treatment of diabetic vascular complications. Although PKCβ is associated with diabetic vascular dysfunction, role of PKCβ in hypertension remains unclear. Since hypertension is one of most important risk factor for vascular diseases including atherosclerosis, we here examined whether PKCβ is involved in angiotensin II-induced vascular dysfunction. Treatment with angiotensin II (100 nM) for 30 min strongly increased phosphorylation at the PKCβ activation site at S660 in aorta from Wistar rat (n=4). Treatment of rat aorta with angiotensin II (100 nM) for 24 hr increased contractile responses to serotonin (5-HT, Maximum contraction, control vs. Angiotensin II, 92.2±20.0 vs 142.1±20.5%, P<0.05, n=6) or endothelin-1 (ET-1, 157.4±30.6 vs 214.1±24.3%, P<0.05, n=8). In contrast, angiotensin II did not affect vascular contractile response to noradrenaline (n=4). The enhanced contraction induced by angiotensin II was blocked by CGP53353 (1 μM), a PKCβ inhibitor (5-HT, Angiotensin II vs. CGP53353+Angiotensin II,197.8±27.8 vs. 167.4±17.6%, P<0.05, n=4-5). Pretreatment with LY333531 (1 μM), another PKCβ inhibitor, also prevented angiotensin II-induced vascular dysfunction (P<0.05, n=3-4). Next, we generated PKCβ knockout rat using rGONAD. Consistent with the results using PKCβ inhibitors, angiotensin II-induced enhanced contractile responses to 5-HT (122.6±15.6 vs 111.0±16.3%, n=4) and ET-1 (137.9±39.9 vs 139.2±30.1%, n=4) were dismissed in PKCβ knockout rat. We also confirmed that phorbol12-myristate13-acetate (0.05-5 μg/ml), a PKC activator, enhanced vascular contractile responses to 5-HT and ET-1 in dose-dependent manner (P<0.05, n=4). We conclude that long exposure of angiotensin II (24 hr) causes increase in vascular contractile responses to 5-HT and ET-1 and the augmented contraction are due to activation of PKCβ. Our result suggests that PKCβ might be associated with angiotensin II-related vascular dysfunction.
Abstract P128: Sexual Dimorphisms In Perivascular Adipose Tissue In A Novel Mouse Model Of Diet-Induced Metabolic Syndrome.
Authors: Tran Vivian , Brettle Holly , Diep Henry , Dinh Quynh Nhu , Sobey Christopher G , Drummond Grant , Vinh Antony , Jelinic Maria
Published: September 2022

Metabolic syndrome (MetS) is a complex multifactorial disease that increases cardiovascular risk. Vascular dysfunction associated with MetS is strongly influenced by perivascular adipose tissue (PVAT) which acts as a reservoir for immune cells. Sexual dimorphisms of PVAT pathophysiology are poorly understood due to a lack of clinically relevant animal models of MetS that show metabolic disturbances in males and females. Using a new diet-induced model of MetS pathophysiology in both sexes, we characterised the aortic leukocyte profile and determined PVAT influence on vascular function. Six-week-old male and female C57BL/6 mice were fed either a high-fat diet (43% kcal) with high sugar and salt (10% high fructose and 0.9% NaCl; HFSS) in drinking water, or normal control diet (NCD) for 10 weeks. Physiological parameters were measured fortnightly. At endpoint, aorta with PVAT intact was isolated for leukocyte enumeration (flow cytometry) or for
in vitro
vascular reactivity (± PVAT; wire myography). Weight gain (NCD M: 29.3±0.5 g vs. MetS M: 37.5±1.0 g; NCD F: 23.6±0.7 g vs. MetS F: 28.9±1.2 g), fasting blood glucose (NCD M: 9.5±0.4 mmol/L vs. MetS M: 11.3 mmol/L; NCD F: 8.9±0.3 mmol/L vs. 10.4±0.5 mmol/L), blood cholesterol (NCD M: 125.0±3.0 mg/dL vs. MetS M: 177.5±6.5 mg/dL; NCD F: 96.7±2.9 mg/dL vs. MetS F: 130.7±3.4 mg/dL) and systolic blood pressure (NCD M: 121.0±1.8 mmHg vs. MetS M: 132.0±2.1 mmHg; NCD F: 122.3±2.2 mmHg vs. MetS F: 133.6±2.7 mmHg) were increased in both sexes of HFSS-fed mice (P<0.05 n= 12; vs. NCD) confirming that the HFSS model is a clinically relevant diet-induced model of MetS. Despite comparable metabolic disturbances in both sexes, aortic inflammation was only observed in males (increased pro-inflammatory monocytes (Ly6C
hi+
; M: 3.3±0.5 x 10
3
cells/aorta vs F: 1.6±0.4 x 10
3
cells/aorta) and neutrophils (Ly6G
+
; M: 0.9±-0.2 x 10
3
cells/aorta vs F: 0.3+/0.1 x 10
3
cells/aorta); P<0.05 vs. HFSS-fed female. MetS altered PVAT and vascular function in both sexes, but with contrasting effects. PVAT blunted endothelium-dependent relaxation in HFSS-fed males (P<0.05 vs. NCD male) and in NCD-fed females (P<0.05 vs. HFSS females). The data indicates that sexual dimorphisms in PVAT impacts MetS-induced aortic inflammation and endothelial dysfunction.
Abstract P131: Abnormal Blood Pressure Fall During Hyperventilation Maneuver In Postural Tachycardia Syndrome
Authors: Urechie Vasile , Rigo Stefano , Shibao Cyndya A , Okamoto Luis E , Gamboa Alfredo , Biaggioni Italo , Wahba Amr , Elkholey Khaled , Giesecke Morgane , Muldowney James , Mohr Abigail E , Diedrich Andre
Published: September 2022

Postural Orthostatic Tachycardia Syndrome (POTS) might have altered endothelial function, causing abnormal vasomotion in the body and in cerebral vessels. Non-cardiac symptoms experienced by these patients suggest that they may be especially sensitive to acute reduction of arterial CO
2
concentration during hyperventilation (HV). We hypothesized HV induces more significant blood pressure (BP) fall with compensatory higher HR increase in POTS. Subjects previously diagnosed with POTS and gender-matched controls were enrolled. ECG, finger BP, and EtCO
2
were continuously recorded during hyperventilation (1breaths/sec) over 30 seconds. Hemodynamic parameters were computed in absolute and change. We studied 14 POTS and 15 controls matched for age (29
±
2 vs 30
±
3; p=0.63) and BMI (24.1±0.8 vs 23.0±0.7; p=0.34). At baseline mean HR (79±4 vs 70±3 bpm, p=0.12) was comparable and SBP was higher (113±3 vs 105±2 mmHg, p<0.05) compared to controls. EtCO
2
values were comparable at baseline (35±1 vs 38±1 mmHg, p=0.14) and at the end of the maneuver (20±1 vs 21±1 mmHg, p=0.58). POTS had greater SBP drop (-27±2 vs -19±2 mmHg, p<0.05) and a comparable HR increase (34±3 vs 29±2 bpm, p=0.11), despite POTS reached a higher absolute maximal HR (113±4 vs 99±2 bpm, p<0.05) [Figure]. Recovery time for HR (31±4 vs 34±4 sec, p=0.68) and SBP (27±7 vs 18±4 sec, p=0.42) were similar. POTS have greater SBP drop with less adequate compensatory HR increase during HV. These results suggest that vascular responsiveness to hypocapnia is especially strong in POTS and it can contribute to orthostatic stress and symptoms.
Abstract P136: Exploring The Interactions Between Rho-related BTB Domain Containing 1 (RhoBTB1) And Phosphodiesterase 5 (PDE5) To Identify Novel Therapeutic Candidates To Counter Hypertension
Authors: Kumar Gaurav , Wu Jing , Brozoski Daniel T , Lu Ko-Ting , Sigmund Curt D
Published: September 2022

Protein-protein interaction studies have provided valuable insights to understand fundamental mechanisms in cells. We previously identified that PDE5 is a binding partner of RhoBTB1 and stringently regulates PDE5 activity in vascular smooth muscle (VSM). This in turn regulates the nitric oxide response of VSM and consequently blood pressure. We showed that RhoBTB1 can function as an adapter for Cullin-3 (CUL3) E3 ring ubiquitin ligase complex delivering PDE5 for ubiquitination and proteasomal degradation. In order to identify other RhoBTB1 binding partners, we assessed the binding interface of RhoBTB1 which binds to PDE5 with the notion that the same region will bind to other RhoBTB1-CUL3 substrates. Full-length RhoBTB1 was truncated into its component domains, separately (GTPase, BTB1, BTB2, and C-terminal (CT)) and in combination (e.g. B1B2C) domains. HEK293 cells were co-transfected with epitope-tagged PDE5 and RhoBTB1 domain constructs, and their interaction was analyzed by co-immunoprecipitation (Co-IP). Interestingly, the CT domain itself was essential for the interaction between RhoBTB1 and PDE5 but was not sufficient on its own to bind to PDE5. This was supported by our finding that the domains lacking CT region could not bind with PDE5. The Co-IP data was further corroborated by immunofluorescence and proximity labeling assay (PLA). Our data suggest that RhoBTB1 could bind to PDE5 utilizing its BTB1-BTB2-CT domains. Moreover, protein stability and ubiquitination studies revealed that the BTB1-BTB2-CT domain is sufficient for CUL3-dependent proteasomal degradation. Molecular approaches including site-directed mutagenesis are being explored to investigate potential “hotspots” underlying the RhoBTB1-PDE5 interaction. We are currently utilizing ascorbate peroxidase (APEX2) fusion system involving BTB1-BTB2-CT domain as “bait” to identify unknown interacting partner in RhoBTB1-Cullin3 axis. Preliminary experiments have been performed to validate the system. For example, BTB1-BTB2-CT-APEX2 can bind to PDE5. These studies are designed to assess if there are other proteins which are involved in the regulation of blood pressure in VSM which may become novel therapeutic candidates against hypertension.
Abstract P206: Are Home Blood Pressure Devices Accurate? A Systematic Review Of The Evidence
Authors: Hiremath Swapnil
Published: September 2022

Background:
Out of office blood pressure (BP) measurements, especially home BP monitoring, are recommended for diagnosis and management of hypertension by many societies including by the Canadian Hypertension Education Programme (CHEP). Though validation protocols exist for individual monitors, little data exists on the real world accuracy of home BP monitors, in actual use. We undertook this systematic review to estimate the accuracy of home BP monitors as reported in the literature.

Methods:
We undertook a literature search of MEDLINE and EMBASE from 1946 and 1947 until April 2015 respectively. We included studies that evaluated the accuracy of home BP devices against a mercury sphygmomanometer considered as the gold standard. Two reviewers independently selected studies, extracted data and assessed quality. Disagreements between the two reviewers were resolved by a third author.

Results:
Our search revealed 797 non-duplicate citations. After applying selection criteria, nineteen studies, involving 4954 patient-devices (median 91, interquartile range 69, 489) were included in the systematic review. The reported inaccuracy of home BP monitors compared to mercury sphygmomanometer ranged from 10% to 72% for systolic BP, with each study using different thresholds for inaccuracy. The absolute mean difference for systolic BP between home BP monitor and the standard ranged from 2.4 mm Hg to 10.4 mm Hg and for diastolic BP from 1 to 8.7 mm Hg.

Conclusion:
The existing literature reports a relatively high degree of inaccuracy in home BP monitors being used. Data is limited by varying definitions being used for reporting inaccuracy. Consideration should be given to standardised definitions of accuracy, and real world monitoring of accuracy as home BP monitors use for clinical decision making becomes widespread. Research into predictors of inaccuracy is also necessary.
Abstract P210: Mir-204 In Endothelial Cells Contributes To The Development Of Hypertension
Authors: Liu Jing , Guenther Maya , Liu Yong , Wu Jing , Liu Xuanchen , Usa Kristie , Pandey Rajan , Markson Ian , Geurts Aron M , Miller Sheldon , Liang Mingyu
Published: September 2022

Global knockout of microRNA miR-204 leads to significant attenuation of hypertension in mice treated with uninephrectomy, angiotensin II (AngII), and a high-salt diet. We hypothesize that miR-204 in endothelial cells (EC) contributes to hypertension. We developed
Mir204
fl/fl
mice and crossed them with VE-Cadherin-Cre mice. The loss of miR-204 was confirmed in isolated endothelial cells (1 ± 0.15, 0.41 and 0.13 ± 0.05, fold in EC-
Mir204
+/+
, EC-
Mir204
+/-
, and EC-
Mir204
-/-
mice, respectively; n=4, 1, and 4). AngII and 4% NaCl diet increased systolic blood pressure (SBP) by up to 40-50 mmHg in both male and female EC-
Mir204
+/-
and WT littermates (n=11 and 17). The SBP was significantly decreased by 20-30 mmHg in EC-
Mir204
-/-
mice (P < 0.05, n=10). Vascular function was assessed by pressure myograph in basilar artery following the Ang II and 4% NaCl diet treatment. In EC-
Mir204
-/-
male mice, the wall thickness (21.5 ± 0.1 vs 30 ± 3.1μm, n=7 and 2) and media/lumen ratio (0.2 ± 0.01 vs 0.51 ± 0.09, n=7 and 2) was significantly reduced compared with EC-
Mir204
+/+
mice. Endothelial-dependent vasodilation (response to Ach) was lost in EC-
Mir204
+/+
mice but largely preserved in EC-
Mir204
-/-
mice (P= 0.007). Renal interstitial fibrosis was not influenced by the miR-204 KO in endothelial cells (Cortex: 0.27 ± 0.04, 0.18 ± 0.01 %; Outer medulla: 0.18 ± 0.05, 0.19 ± 0.05 % in EC-
Mir204
+/+
and EC-
Mir204
-/-
male mice, respectively; n=3 and 2). Urinary albumin excretion was increased during the two-week treatment. However, the albuminuria was not significantly affected by the knockout of miR-204 in endothelial cells (2.65 ± 1.13, 3.41 ± 0.86, and 2.86 ± 0.85 mg/24h in EC-
Mir204
+/+
, EC-
Mir204
+/-
, and EC-
Mir204
-/-
mice, respectively; n=9, 11 and 6). Glomerular filtration rate did not differ among the three groups at baseline or study termination (Baseline: 188.3 ± 30.1, 223.1 ± 34.9, and 185.3 ± 21.3 μl/min, n=4, 2 and 5; Termination: 203.8 ± 22.1, 150.7 ± 30.3, and 193.9 ± 42.5 μl/min, n=6, 2 and 5 in EC-
Mir204
+/+
, EC-
Mir204
+/-
, and EC-
Mir204
-/-
mice, respectively). The findings suggest that miR-204 in endothelial cells plays an important role in the development of hypertension associated with impaired endothelial-dependent vasodilation and vascular remodeling.
Abstract P213: Bridging The Gap In Hypertension Outcomes In Rural United States: A 6-month Report From A Pharmacist-run Hypertension Clinic In An Appalachian University Teaching Hospital
Authors: Ezeh Ebubechukwu , Messer Brittany , Wang Xiaoliang , Mansoor Kanaan , Thompson Ellen
Published: September 2022

Background:
Hypertension (HTN) constitutes a significant risk factor for cardiovascular diseases, including stroke and myocardial infarction. According to CDC, HTN rate is 10% higher in rural areas than in most urban areas. Rural Appalachia has even higher HTN rates (>10% above the national rate). Optimization of rural health care is critical in improving overall outcomes of HTN. We, thus, compared a 6-month trend in BP control pre and post-referral of patients to a pharmacist-led BP clinic.

Methods:
This was a prospective study carried out in the HTN clinic of an Appalachian teaching hospital. The analysis was restricted to hypertensive patients aged ≥18 years. Interventions included medication management and health promotion. Average and upper limits of BP and the number of office visits in both periods were collected. Paired T-test was used for analysis. A p-value <0.05 is considered to be statistically significant.

Results:
Of the 51 persons analyzed, 38 were eligible for inclusion. The average systolic BP demonstrated a significant decline from 160s to 140s mm Hg (p-value <0.0001; 95% confidence interval -27.83 to -14.75; SEM 3.228). DBP showed a drop from around 85 to 80 mm Hg (p-value 0.0039; 95% CI -9.683 to -2.002; SEM of 1.895). The upper limits of SBP before and after referral also reached statistical significance (p-value 0.0008; CI -24.43 to -6.976; SEM 4.303). The BMI did not reach statistical significance. Patients visited the clinic almost twice as much after referral within the 6-month period that was studied (1.27 vs 2.29).

Conclusions:
Overall, pharmacist-led BP control may potentially play a huge role in improving the outcomes of HTN in resource-limited, rural areas.
Abstract P214: Disparities In Hypertension Medication Adherence Across Sexual Orientations
Authors: Hyder Syed , Schoenl Samantha A , Kesiena Onoriode
Published: September 2022

Objective:
The LGBTQ community faces unique psychosocial stressors and barriers in access to care. This study seeks to characterize the difference in hypertension (HTN) medication adherence between LGBTQ and straight men and the difference between LGBTQ and straight women.

Methods:
This cross-sectional study examines 1,905 self-identifying men and 2,349 women, who selected “gay” or “bisexual”, and 66,938 men and 76,464 women, who selected “straight”, for sexual orientation in response to the National Health Interview Survey (NHIS) conducted by the Center for Disease Control and Prevention (CDC) from 2016-2021. Other sexual orientations included under the umbrella term, LGBTQ, were not listed as options on the NHIS survey. Unpaired t-test was used to compare the study groups and linear regression was used to calculate slope (m) of the trendline. Statistical analysis was weighted for population size.

Results:
Gay/bisexual men [x = 70.1%, IQR 3.75%] had significantly lower rates [p < 0.001] of HTN medication adherence compared to straight men [x = 78.1%, IQR 5.5%]. The difference in HTN medication adherence between gay/bisexual women [x = 72.2%, IQR 13.1%] and straight women [x =82.4%, IQR 2.80], was also statistically significant [p = 0.014]. Adherence in both gay/bisexual and straight groups trended upward since 2016; however, the upward trend was steeper for straight men [m= 1.87] and women [m= 0.81] compared to gay/bisexual men [m= 1] and women [m= 0.12].

Conclusion:
The findings of this study display significantly lower rates of HTN medication adherence between males and females who identify as gay/bisexual compared to those that identify as straight. Since 2016, the gap in medication adherence between gay/bisexual and straight has continued to widen. As medical systems strive to reduce health disparities across various marginalized groups, further research is needed to delineate potential sources for this difference and devise solutions.
Abstract P220: Effects Of Orthostatic Hypotension On Intensive Blood Pressure Treatment With Respect To All-cause Mortality: An Individual-level Meta-analysis
Authors: Juraschek Stephen P , Hu Jiun-Ruey , Cluett Jennifer L , Ishak Anthony M , Mita Carol , Lipsitz Lewis A , Appel Lawrence J , Beckett Nigel S , Coleman Ruth L , Cushman William C , Davis Barry R , Grandits Greg , Holman Rury R , Miller Edgar R , Peters Ruth , Staessen Jan A , Taylor Addison A , Wright Jackson T , Mukamal Kenneth J
Published: September 2022

Background:
In a recent individual level meta-analysis, intensive versus standard blood pressure (BP) treatment reduced participants’ risk of orthostatic hypotension (OH). Whether OH modified the relationship between intensive treatment and risk of cardiovascular disease (CVD) or death is unknown.

Methods:
We performed an individual participant data meta-analysis, updating a previous systematic review of MEDLINE, EMBASE, and CENTRAL databases through May 13, 2022, which included randomized trials of BP pharmacologic treatment (more intensive BP goal or active agent) on CVD or death. CVD events were adjudicated and included coronary heart disease, stroke, and congestive heart failure. OH was defined as a drop in SBP ≥20 mmHg and/or DBP ≥10 mmHg after changing positions from sitting to standing. Ultimately, 8 trials were identified with OH and outcomes data. Effects were examined overall and by trial type (BP goal or active agent), using Cox proportional hazard models adjusted for age and sex.

Results:
There were 27,974 participants followed for median of 4 years (mean age 69.5±10.7 years; 48.5% female; 8.6% with OH). Baseline OH was associated with a higher risk of CVD or death (HR 1.24; 95% CI: 1.10, 1.39). More intensive BP treatment or active therapy lowered risk of CVD or death among those without OH at baseline (HR 0.82; 95% CI: 0.76, 0.88) and with OH at baseline (HR 0.79; 95% CI: 0.63, 0.98). Effects did not differ by baseline OH (
P
-interaction 0.78) (
Table
).

Conclusion:
While baseline OH was associated with CVD or death, it did not increase the risk of CVD or death from intensive treatment. OH prior to the initiation of intensive therapy should not be viewed as a reason to avoid BP treatment.
Abstract P233: Cardiac Regulator Of G Protein Signaling-4 Upregulation Protects Against FFAR3-mediated Inflammation And Sympathetic Hyperactivity
Authors: Cora Natalie , Carbone Alexandra , Suster Malka , Borges Jordana , Ferraino Krysten , Desimine Victoria , Maning Jennifer , Lymperopoulos Anastasios
Published: September 2022

Background:
Free fatty acid receptor (FFAR)-3 is a Gi/o protein-coupled receptor (GPCR) mediating many cellular effects of short-chain fatty acids like propionate. It inhibits cyclic 3’,5’-adenosine monophosphate (cAMP) synthesis and promotes sympathetic nervous system (SNS) activity and norepinephrine (NE) release. Regulator of G protein Signaling (RGS)-4 deactivates (terminates) Gi/o- and Gq-protein signaling. Cardiac RGS4 protects the heart against arrhythmogenesis via calcium signaling attenuation and against pressure overload-induced hypertrophy.

Study objective:
We examined whether RGS4 regulates FFAR3 signaling and function in cardiac myocytes and sympathetic neurons in vitro, as well as in mice post-pressure overload in vivo.

Methods & Results:
siRNA-mediated RGS4 depletion in H9c2 cardiomyocytes enhances propionate-induced cAMP lowering, Giα protein subunit activation, and upregulation of pro-inflammatory mediators [p38 MAPK, interleukin (IL)-6 & IL-1β, transforming growth factor (TGF)-β]. On the other hand, RGS4 overexpression in sympathetic neuron-like Neuro-2A cells significantly reduces propionate/FFAR3-mediated NE release. In vivo, in C57/B6 mice undergoing transverse aortic constriction (TAC) to induce pressure overload (increased afterload), cardiac RGS4 was found upregulated, at both the mRNA and protein levels, in post-TAC mice compared to sham-operated controls. This was accompanied by markedly reduced propionic acid-dependent FFAR3 signaling in isolated membranes from the post-TAC hearts. Finally, although initially (24 hours post-TAC) elevated, compared to sham (6.6
+
0.4 nmol/g of tissue vs. 4.1
+
0.7 nmol/g of tissue, respectively; n=5), cardiac NE levels were markedly suppressed at 7 days post-TAC (5.4
+
0.3 nmol/g of heart tissue, n=5).

Conclusions:
RGS4 inhibits cardiac propionic acid/FFAR3 pro-inflammatory, and neuronal propionic acid/FFAR3 NE release-stimulating signaling. Pressure overload upregulates cardiac RGS4 in vivo to protect the heart against FFAR3-mediated inflammation and elevated SNS activity/NE release. Thus, cardiac RGS4 upregulation might represent a cardioprotective mechanism against the chronically increased afterload imposed by systemic hypertension.