bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P056: Sex Differences In Association Between Gut Microbiome And Essential Hypertension Based On Ambulatory Blood Pressure Monitoring
Authors: LAU Kui Kai , VIRWANI Preeti D , QIAN Gordon , HSU Matthew SS , PIJARNVANIT Tommy KKTS , CHEUNG Carman Nga-Man , CHOW Yick Hin , TANG Lok Kan , TSE Yiu-Hei , XIAN Jia-Wen , LAM Shirley Sau-Wing , LEE Crystal , LO Chelsea CW , LIU Roxanna KC , HO Tsi Lok , CHOW Bak Yue , LEUNG Kin Sum , TSANG Hing Wai , LO Emily Kwun Kwan , TUNG Keith TS , IP Patrick , HUNG Ivan Fan-Ngai , LOUIE Jimmy Chun Yu , EL-NEZAMI Hani , HO Joshua WK
Published: September 2022

Background:
Sex differences in the pathogenesis of hypertension exist. Whilst gut microbiota (GM) dysregulation is a novel risk factor of hypertension, studies using shotgun metagenomics sequencing and ambulatory 24-hour blood pressure (BP) monitoring to study the potential sex differences in the associations between the GM and BP are lacking.

Methods:
We recruited 241 asymptomatic Hong Kong Chinese (113 male, 128 female, mean age 54±6), who were not on anti-hypertensive agents, and studied their GM composition using shotgun sequencing. 24-hour ambulatory BP was recorded. GM-mediated short chain fatty acids (SCFAs) were quantified in stool and plasma by GC-MS. Statistical analysis was performed under covariate-adjusted models including age, sex, BMI, smoking, sodium-intake, fatty liver and menopause status.

Results:
Based on 24-hour BP, 36% (87 of 241) of the study population was hypertensive. Males had a higher mean BP than females (127±13/81±10mmHg vs. 117±12/71±8mmHg, p<0.001). GM β-diversity was significantly different in hypertensive vs. normotensive individuals, driven by females. A significant enrichment of
Ruminococcus gnavus
was observed in the hypertensive group whilst
Oscillibacter sp. CAG:241
and
Gemmiger formicilis
was enriched in the normotensive group. These differences remained significant in females with no significant differential species found in males. Additionally,
Dorea formicigenerans
was negatively associated whilst
Clostridium bolteae
and
Bacteroides ovatus
were positively associated in female-specific hypertension. Repeated cross-validation machine-learning demonstrated microbial features were more predictive of hypertension in females than males, and the addition of microbial features to clinical features (age, BMI) improved the model’s prediction accuracy from 0.69 to 0.84 in females. Total plasma SCFAs and propionic acid were independent predictors of systolic and diastolic BP in females but not males. GM β-diversity was also significantly associated with total SCFAs and propionic acid levels.

Conclusion:
GM dysregulation appears to be more strongly associated with hypertension in females, which may be mediated through circulating plasma SCFAs.
Abstract P061: Effect Of Short Chain Fatty Acid Butyrate On Cardiovascular Parameters In The Zebrafish Larvae
Authors: Sree Kumar Hemaa , Wisner Alexander S , Schiefer Isaac T , Zubcevic Jasenka
Published: September 2022

Introduction:
Short-chain fatty acids (SCFAs) such as butyrate are produced by microbial fermentation of dietary fiber in the gut. While the effects of butyrate on cardiovascular function have previously been described in rodent models, no studies to date have investigated butyrate effects on cardiovascular function in zebrafish (ZF). ZF have become a widely used including in cardiovascular and gut dysbiosis research, and bacterial communities from adult ZF intestines can reportedly synthesize butyrate. Here, we investigated chronotropic and vasoactive effects of acute administration of butyrate in ZF larvae.

Methods:
ZF larvae (7-9 dpf) were anaesthetized with tricaine by immersion (MS-222, 0.3mM in larvae water). For measurements of heart rate (HR), either butyrate (40mM final concentration, ph=7.5) or vehicle (larvae water) was added to the ZF chambers, and HR was recorded with phase contrast microscopy every 10 minutes for an hour. For measurement of vascular reactivity, 30s-long videos of the ZF dorsal aorta were taken before (i.e., at baseline) and after administration of butyrate (40mM final concentration, ph=7.5). Images were exported into ImageJ, and the average width was recorded at several points along the length of the dorsal aorta.

Results:
We observed significantly lower HR in the ZF at 20min following butyrate administration (Mean±SEM: 117±6 bpm) compared to vehicle (Mean±SEM: 131±5 bpm; P<0.05), suggesting a negative chronotropic effect of butyrate. In addition, compared to baseline, we observed a ~20% increase in dorsal aorta width following acute administration of butyrate, suggesting vasodilatory effects.

Conclusion:
Here we show, for the first time, that butyrate can produce similar cardiovascular effects in the ZF as in the rodent models. Thus, ZF may be useful for investigation of gut microbial effects on cardiovascular parameters.
Abstract P075: Functional NOX2 In T Cells Amplifies Salt-Sensitive Hypertension And Associated Renal Damage
Authors: Walton Samuel D , Dasinger John H , Shaw Mary , Burns Emily , Abais-Battad Justine M , Mattson David L
Published: September 2022

Infiltrating T cells in the kidney amplify salt-sensitive (SS) hypertension and renal damage, but the mechanisms are not known. Genetic deletion of T cells (SS
CD247-/-
) or the p67phox subunit of NOX2 (SS
p67phox-/-
) attenuates salt-sensitive hypertension in the Dahl SS rat. We hypothesized that reactive oxygen species (ROS) produced by NOX2 in T cells drive the SS phenotype and renal damage. T cells were reconstituted by adoptively transferring ~10 million splenocytes from the Dahl SS (SS→CD247, n=14), the SS
p67phox-/-
(p67phox→CD247, n=19), or PBS only (PBS→CD247, n=8), into the peritoneum of SS
CD247-/-
rats on postnatal day 5. Animals were instrumented with radiotelemeters at 8 weeks of age. After a week of recovery, blood pressure and albumin excretion were measured while rats were fed a low salt (LS, 0.4% NaCl) diet. Rats were then switched to a high salt (HS, 4.0% NaCl) diet for 21 days, with continuous blood pressure measurements and urine collections every 7 days. On day 22, the rats were euthanized, and their kidneys were perfused and collected for histology and renal immune cell characterization by flow cytometry. At baseline there was no detectable difference in mean arterial pressure (MAP) between groups (124±2 vs 123±2 vs 116±2 mmHg; SS→CD247 vs p67phox→CD247 vs PBS→CD247, respectively). After 21 days of HS, MAP was significantly greater in the SS→CD247 compared to the p67phox→CD247 rats (155±3 vs 144±2 mmHg; p<0.001) and the PBS→CD247 rats (155±3 vs 141±3 mmHg; p<0.001). Indices of renal damage identified no difference in albuminuria between SS→CD247 and p67phox→CD247 on LS (25.8±4.7 vs 23.2±3.2 mg/day). After 21 days of HS, albuminuria was significantly greater in the SS→CD247 (242.8±30.3 mg/day) compared to the p67phox→CD247 (175.5 ± 14.3 mg/day, p<0.03) and PBS→CD247 (135.9±27.8 mg/day, p<0.005). There was no difference between p67phox→CD247 and PBS→CD247 in albuminuria or MAP during LS or HS. The lack of CD3+ cells in the PBS→CD247 but presence of CD3+ cells in rats receiving the T cell transfer demonstrated the effectiveness of the adoptive transfer. These results indicate that ROS production by T cells participates in the amplification of salt-sensitive hypertension and renal damage.
Abstract P084: Exploring The Role Of Necroptosis In Hypertension
Authors: O'Donnell Joanne A , Dinakis Evany , Rhys-Jones Dakota , Muralitharan Rikesih M , Marques Francine Z
Published: September 2022

Objective:
Altered immune cell activation plays a key role in promoting hypertension. How the immune system becomes activated during hypertension is unknown. A possibility is necroptosis, a newly described form of cell death that alerts the immune system to dying cells, causing inflammation. Using

Mlkl
-/-

mice that are genetically unable to undergo necroptosis, we explored the role of necroptosis in inflammation or blood pressure regulation during hypertension. We also examined whether Ang-II sensitizes human and murine cells to necroptosis.

Methods:
Monocytes were sorted from peripheral blood mononuclear cells of 3 normotensive and 3 untreated hypertensive participants diagnosed with 24h BP monitoring. Primary murine cardiomyocytes and fibroblasts were obtained from 3 litters of neonatal C57BL/6J mice. Necroptosis was induced using TNF-alpha, SMAC mimetic and zVAD in the presence or absence of Ang-II. Viability of human cells was measured using propidium iodide and in murine cells with CellTitre-Glo Viability Assay. Hypertension was induced in male

Mlkl
-/-

mice and WT littermate controls by implanting minipumps containing Ang-II (0.5mg/kg/day, 28 days; n=6/group). BP was measured weekly by tail-cuff.

Results:
Monocytes from hypertensive participants were not more susceptible to necroptosis than normotensive participants (P=0.79). This was not altered by Ang-II treatment in normotensive (P=0.98) or hypertensive participants (P=0.92). Moreover, Ang-II did not alter the viability of primary murine fibroblasts
in vitro
(P=0.79). Mouse cardiac fibroblasts were unable to undergo necroptosis. In vivo, there was no difference in BP of Ang-II-treated

Mlkl
-/-

and WT control mice at 4 weeks (P=0.87).

Conclusions:
We show that monocytes from hypertensive participants were not more susceptible to necroptosis, compared to normotensive participants. Furthermore, Ang-II did sensitize mouse cardiac fibroblasts or human monocytes to necroptosis. No changes in BP were observed between Ang-II treated WT and MLKL knockout mice, suggesting necroptosis does not impact BP regulation. Further analyses are being performed to determine if inflammation or fibrosis are altered when necroptosis is inhibited.
Abstract P096: Association Of Maternal Blood Pressure With Offspring Blood Pressure In A Hypertensive Non-Human Primate Model
Authors: Vincent Carol , Chen Ashton , Kavanagh Kylie , South Andrew M
Published: September 2022

Background:
Mechanisms behind primary HTN-related organ damage are not well understood and animal models are lacking. The spontaneous HTN vervet monkey (
Chlorocebus spp.
) is a potential model for studying HTN development across the lifespan.

Methods:
This is a prospective cross-sectional study of
Chlorocebus spp.
maternal-offspring dyads. We identified 10 non-pregnant HTN mothers based on prior sedated BP measurements and randomly selected a single offspring to include. We matched HTN dyads by age and offspring sex 2:1 with maternal normotensive dyads. We defined HTN by the 2017 ACC/AHA Guideline criteria. We obtained sedated BP, weight, and labs on all dyads. Our exposures were presence of maternal HTN and BP in mothers. Our outcome was offspring BP. We estimated associations between the exposures and outcome with adjusted generalized linear models.

Results:
Maternal age ranged 9.6—19.7 years and offspring age ranged 1.6—8.7 years. Maternal systolic BP was associated inversely with offspring systolic BP (
β
-0.44 mmHg, 95% CL -0.87 to -0.01) and maternal HTN was associated with lower diastolic BP (
β
-16.1 mmHg, 95% CL -30.7 to -1.6).

Conclusion:
In this vervet monkey HTN model, we observed an intriguing association of maternal HTN with lower offspring BP. The variability of the vervet monkeys’ BP over time could be related to environmental factors, and BP alone may not be the most reliable measure of cardiovascular risk in this group. Future steps include investigating association of biochemical markers and renin-angiotensin-aldosterone system components with BP and target organ damage in this model.
Abstract P101: AT1-AA Induced Hypertension During Pregnancy Decreases Maternal Cerebral Blood Flow Autoregulation Postpartum
Authors: Campbell Nathan , Strong Luke , Fang Xing , Herrock Owen , Fitzgerald Sarah , Turner Ty , Deer Evangeline M , Amaral Lorena M , Roman Richard J , Lamarca Babbette B
Published: September 2022

Preeclampsia (PE), new-onset hypertension during pregnancy, is the leading cause of morbidity and mortality for the mother and is associated with multi-organ dysfunction including the brain. Women with PE have activated B cells producing agonistic autoantibodies to the angiotensin II type 1 receptor (AT1-AA) which remain elevated in maternal circulation up to 8 years postpartum (PP); therefore, we believe it to play a role in PE pathophysiology and the increased incidents of cardiovascular disorders PP. Although we have shown at AT1-AA infusion into pregnant rats results in elevated mean arterial pressure (MAP), impaired cerebral blood flow (CBF) autoregulation, and reduced pup weight, we don’t know the effects of AT1-AA on MAP and CBF hemodynamics in the PP period. We hypothesize that AT1-AA induced hypertension during pregnancy will cause maternal hypertension and impaired maternal CBF PP. To test this hypothesis, AT1-AA (1:40) was infused on gestational day (GD) 14 via a mini-osmotic pump. Dams were allowed to deliver; pup weights were recorded within 12 hours. PP dams were aged to 9 weeks after birth and one group of AT1-AA PP dams received a second infusion of AT1-AA (1:80), to mimic the levels of AT1-AA seen in PP PE women. At 10 weeks PP, maternal MAP was measured and at 12 weeks PP, CBF autoregulation was measured by laser Doppler flowmeter. At PP week 12, MAP was 118±3 mmHg in NP (n=4), 119±2 mmHg in AT1-AA (n=5), and 126±5 in AT1-AA + AT1-AA (1:80) PP (P<0.05, n=5). CBF increased by 43±8% (P<0.05, n=2) in rats with AT1-AA during pregnancy and by 37±9% (P<0.05, n=4) in rats with AT1-AA during pregnancy and (1:80) PP in response to increased MAP from 100 to 140 mmHg, versus only 6±2% (n=3) in normal pregnant controls. In conclusion, AT1-AA during pregnancy causes sustained changes in CBF hemodynamics PP. Sustained maternal AT1-AA PP also causes increased blood pressures long-term in association with impaired CBF. These data indicate that perinatal and PP AT1-AA cause long-term cardiovascular and cerebrovascular consequences for PE women.
Abstract P110: Differential Gene Expression Associated With Ccn1-positive Renal Cells In Murine Hypertensive Models
Authors: McDermott Justin G , Creed Heidi , Navaneethabalakrishnan Shobana , Goodlett Bethany , Rutkowski Joseph M , Mitchell Brett M
Published: September 2022

Hypertension affects roughly half of the U.S. population and is characterized by detrimental pro-inflammatory changes that alter renal function and the cardiovascular system over time. The kidney is a major regulator of blood pressure, however, the specific contributions of renal cell types, including lymphatic endothelial cells (LECs), to hypertension have not been thoroughly explored. Previous work from our lab has shown an increase in renal lymphangiogenesis in response to hypertension and that augmentation of renal lymphangiogenesis can reduce blood pressure. CCN1/Cyr61 is an extracellular matrix protein secreted by multiple cell types, particularly LECs and fibroblasts, which contributes to integrin signaling pathways. CCN1 is also known to be involved in lymphangiogenesis, and has previously been implicated in GWAS studies of hypertension. Given these connections, we performed single cell RNA sequencing using angiotensin II and salt sensitive mouse models of hypertension, which were analyzed separately as well as aggregated into one hypertensive group. Kidneys were turned to a single cell suspension and separated for CD31+/Podoplanin+ cells meant to better represent LECs, which are a small fraction of renal cells. Sequencing was performed and the gene reads were run through the Cell Ranger, Seurat, FGSEA, and Slingshot pipelines, then CCN1+ cells were isolated digitally and analyzed separately. In the AngII model, a total of 60 genes (28 up, 32 down) were differentially expressed (p<0.01) between the renal CCN1+ cells in the hypertensive mice compared to controls, while the salt sensitive model had a total of 331 differentially expressed genes (305 up, 26 down). When the hypertension groups were merged, there were a total of 600 differentially expressed genes (503 up, 97 down) with GSEA showing 48 pathways with enrichment scores >3 or <-3. Additionally, the cellular profile of CCN1+ cells shifted, branching out from a fibroblast-like population to other cell types such as endothelium and tubular epithelium. Overall, the expression of CCN1+ renal cells demonstrates notable changes in response to hypertension in murine models and encourages further study of the mechanisms behind CCN1’s role in the development and maintenance of hypertension.
Abstract P114: Differential Expression Of Angiotensin Ii Receptors In Aorta Of Ela2ko Mice With Abdominal Aortic Aneurysm.
Authors: Couto Ariel E , Mestriner Fabiola , D Avila Mesquita Carolina , Corsi Carlos , Flora Vinicius , Franco Francisco Daniely , Campos Ligia C , Rosales Roberta R , Vasconcelos Jocianny L , Polonis Katarzyna , Ribeiro Maurício S , Becari Christiane
Published: September 2022

Elastase-2 (ELA-2) enzyme generates angiotensin-II (AngII) in arteries and is implicated in abnormal vascular remodeling in animal models.Our group demonstrated previously that ELA-2 knockout (ELA-2KO) mice were less susceptible to develop an aorta dilation in a response to AngII infusion. In this study, we hypothesize that susceptibility to an aorta dilation and abdominal aortic aneurysm (AAA) is mediated by AngII receptors (AT1 and AT2).

Methods:
Male wild type (C57bl/6, WT) and ELA-2KO (CELA-2aTm1Bdr) mice were treated with either saline or AngII (2.28g/g/min) for twenty-eight days by subcutaneously mini-pumps infusion. Mice were divided into four groups: WT treated with saline (WT+SAL, n=5), ELA-2KO treated with saline (ELA-2KO+SAL, n=5), WT treated with AngII (WT+AngII,n=5), and ELA-2KO treated with AngII (ELA-2KO+AngII, n=5). Protein expression of AT1 and AT2 receptor, CD68, and α-actin was measured by immunofluorescence in formalin-fixed, paraffin-embedded aorta tissue. ANOVA and Kruskal Wallis tests were used to compare the average fluorescent intensity between groups.

Results:
AngII treatment was associated with increased AT1 receptor expression in a WT group, but not with AT2 receptor expression (vs. WT+SAL,p=0.06 and p=0.31, respectively). The ELA-2KO group showed no difference in AT1 receptor expression in a response to AngII infusion compared to a ELA-2KO+SAL group (p=0.81). While ELA-2KO+Ang II mice had lower AT2 expression compared to ELA-2KO+SAL (p=0.05). The α-actin expression was similar in all groups. WT+AngII group expressed more CD68 expression in a response to an AngII treatment that was increased in a WT group (p=0.03), but not in a ELA-2KO group.

Conclusion:
Our data demonstrate that Ang II treatment did not increase an expression of AT1 receptor in ELA-2KO, but was associated with a downregulation of AT2 receptors. Taken together, this suggests That ELA-2KO mice resistance to developing an aorta dilation/AAA is related to downregulation of Ang II receptors. Our results also highlight the importance of the local/tissue response in the pathophysiology of AAA.
Abstract P115: Oxidized Ldl Potentiates Angiotensin Ii-induced gαQ Activation Through The At1-lox1 Receptor Complex In Kidney Cells
Authors: HUANG YIBIN , Yamamoto Koichi , Rakugi Hiromi
Published: September 2022

Introduction:
In this study, we performed cell experiments to investigate how the co-treatment of oxLDL and AII activates AT1 particularly by focusing on Gαq-dependent pathway.

Methods:
We used CHO-cells stably transfected with AT1 (CHO-AT1), LOX-1 and AT1 (CHO-LOX-1-AT1) and two rat kidney cells (NRK49F and NRK52E). Activation of Gαq and the subsequent induction of Ca influx were quantified by accumulation of IP1 and Fura-2 AM assay, respectively. We also assessed protein expression of αSMA as a molecular marker of epithelial mesanchymal transition (EMT) after 3 days of the indicated stimulation in NRK49F and NRK52E.The proliferative activity was measured using BrdU assay in NRK49F.

Results:
The activation of Gαq was not observed by oxLDL treatment alone but was enhanced by the co-treatment of oxLDL and AII compared to AII alone in CHO-LOX-1-AT1 but not in CHO-AT1. Ca influx was induced not by oxLDL alone but by the co-treatment of oxLDL and a low concentration of AII that was insufficient to induce Ca influx when treated alone in CHO-LOX-1-AT1. We also found that the activation of Gαq and the induction of Ca influx were detected by the co-treatment of oxLDL and AII but not by single treatment in rat kidney cells. These phenomenons were inhibited by ARB, a Gq inhibitor or the siRNA knockdown of AT1 or LOX-1. Furthermore, the EMT was induced only by the combination treatment compared to the single treatment. These phenomenons were inhibited either by ARB or the Gq inhibitor. The combination treatment enhanced the proliferative activity of kidney fibroblasts compared with single treatment, that was inhibited by ARB, the Gq inhibitor or the siRNA knockdown of AT1 or LOX-1..

Conclusions:
These findings suggest the synergistic role of the AT1-LOX-1 interaction whereby oxLDL potentiates Ang II-induced Gαq signaling, leading to the enhancement of renal injury-associated cellular response.
Abstract P127: Systemic Immune-Inflammatory Index (SII) In Hypertensive Adults
Authors: DeSouza Christian S , Garcia Vinicius P , Greiner Jared J , Stauffer Brian L , DeSouza Christopher A
Published: September 2022

Aside from age, hypertension is the most predominant cardiovascular disease risk factor worldwide. In the United States, ~45% of the adult population have blood pressure higher than 130/80 mmHg. Hypertension is associated with increased risk of myocardial infarction, heart failure, stroke and vascular disease. Systemic immune-inflammation index (SII) is a novel biomarker of systemic inflammation and immunity. Clinical interest in SII has intensified due to its association with major cardiovascular events and prognostic value in cardiovascular outcomes. Indeed, the index has been shown to be a strong prognostic marker for coronary artery disease, heart failure, cardiac death and stroke. It is currently unknown whether SII is elevated in adults with elevated blood pressure. We tested the hypothesis that hypertension is associated with an elevated SII, independent of other cardiometabolic risk factors and/or disease. Fifty-six adults (age range: 43 to 66 years) were studied: 22 normotensive (NT: 10M/12F; age 57
+
1 yr; BMI: 27.7
+
1.0 kg/m
2
; BP: 120/75
+
1/1 mmHg) and 34 hypertensive (HT: 23M/11F; age 57
+
1 yr; 28.4
+
0.6 kg/m
2
; BP 140/85
+
1/1 mmHg). All subjects were sedentary, non-smokers, normolipidemic, normoglycemic, non-medicated, and free of overt cardiometabolic diseases. SII was calculated as total peripheral platelets count x neutrophil-to-lymphocyte ratio. SII was significantly higher (~20%) in the HT (479.6
+
25.0) vs NT (402.5
+
27.8) group. There was no significant sex by blood pressure interaction with SII. SII was similar and equally elevated in HT males (487.7
+
34.7) and females (462.7
+
28.7) than NT males (364.4
+
30.8) and females (425.9
+
39.9). SII was independently associated with systolic (r=0.30; P<0.05) and mean arterial (r=0.26; P<0.05) blood pressure; SII was not significantly associated with any other anthropometric, hemodynamic or metabolic variable. Hypertension is associated with elevated SII indicative of greater systemic immune-inflammatory activation. Increased immune activation and inflammatory burden contribute not only to atherosclerosis but to progression of hypertension. Given the link between SII and poor prognosis, SII may aid in identifying high risk hypertensive groups.
Abstract P137: Proteomic And Phosphoproteomic Profiling Reveals Structural And Functional Changes In The Abdominal Aorta Of Male Sprague Dawley Rats That Associate With Age-dependent Hypertension
Authors: Amraei Razie , Lampl Noah , Emili Andrew , Wainford Richard D
Published: September 2022

Aim:
We hypothesized that alterations in the expression and phosphorylation of abdominal aorta protein networks contribute to arterial remodeling in age-dependent hypertension (HTN).

Methods:
Mean arterial pressure (MAP) and blood pressure variability (BPV) were measured by femoral artery cannulation in 3- and 16-month-old (mo) male Sprague-Dawley (SD) rats (N=4/group). Quantitative proteomic/phosphoproteomic analysis was performed on individual abdominal aorta. The differentially expressed/phosphorylated proteins identified at 3 vs. 16-mo (
Abstract P142: Progranulin Controls Vascular Proliferation And Migration Via Regulating Autophagy Flux
Authors: Singh Shubhnita , Bruder Nascimento Ariane , Bruder-Nascimento Thiago
Published: September 2022

Progranulin controls vascular proliferation and migration via regulating autophagy flux Shubhnita Singh, Ariane Bruder-Nascimento, Thiago Bruder-Nascimento Vascular smooth muscle cells (VSMC) play a critical role in the genesis of varied cardiovascular diseases by controlling the integrity of the arterial wall. Progranulin (PGRN) is a protein expressed in multiple cell types, including VSMC, which regulates repair and inflammation. Deficiency in PGRN leads to frontotemporal dementia, but whether such ablation can interfere on VSMC biology is not understood. We tested the hypothesis that PGRN maintains the VSMC homeostasis by regulating autophagy flux. Experiments were performed in aortic VSMC (aVSMC) and arteries isolated from wild-type (PGRN+/+) and PGRN deficient mice (PGRN-/-). PGRN-/- aVSMC showed increase in proliferation and migration measured by Ki67 and PCNA expressions and scratch assay, respectively. An RNA-sequencing in aortae revealed a suppression in autophagy pathway in PGRN-/-. To analyze whether such reduction is associated with impaired autophagy flux, we analyzed the p62 and LC3A/B expression, PGRN-/- aVSMC displayed reduced p62 (2-fold less) and more accumulation of LC3B/A ratio, which was further accentuated by pre-treating the PGRN-/- aVSMC with Bafilomycin A1. Spermidine (autophagy inducer) restored the vascular proliferation and migration, as well as reduces Erk1/2 phosphorylation. In ex vivo experiments, we studied whether impaired autophagy flux in PGRN-/- mice is associated with vascular dysfunction, interestingly we observed that intact aortae from PGRN-/- presented an elevated vascular constriction to norepinephrine (maximal response in mN, PGRN+/+: 4.5 ± 0.4 vs PGRN-/-: 5.3 ± 0.3*, *P<0.05) and thromboxane A2 analogue (maximal response in mN, PGRN+/+: 5.2 ± 0.3 vs PGRN-/-: 6.0 ± 0.5*, *P<0.05), which was abolished by treating the mice with spermidine for 7 consecutive days (4mM in drinking water). Our findings indicate that PGRN regulates the vascular physiology by regulating autophagy flux and propound PGRN deficiency as a risk factor for cardiovascular disease.
Abstract P201: Canagliflozin, Visit-to-visit Blood Pressure Variability, And Risk Of Cardiovascular, Kidney, And Mortality Outcomes: Pooled Individual Participant Data From The CANVAS And CREDENCE Trials
Authors: Fletcher Robert A , Arnott Clare , Rockenschaub Patrick , Schutte Aletta E , Carpenter Lewis , Vaduganathan Muthiah , Heerspink Hiddo J , Neal Bruce , Jun Min , Rodgers Anthony , Perkovic Vlado , Neuen Brendon
Published: September 2022

Background:
Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduce systolic blood pressure (SBP), but whether they affect SBP variability is unknown. There also remains uncertainty about the prognostic value of SBP variability for clinical outcomes.

Methods:
We conducted a post-hoc analysis of the CANVAS Program and CREDENCE trial in individuals with type 2 diabetes at high cardiovascular risk and/or with chronic kidney disease. We used linear mixed models to assess the effect of canagliflozin on SBP variability across 4 study visits over 1.5 years after randomization as measured by standard deviation (SD), coefficient of variation (CV), and variability independent of the mean (VIM). We used multivariable Cox regression models to assess the association of SBP variability with clinical outcomes.

Results:
In 11551 trial participants, canagliflozin modestly lowered SD of SBP variability (-0.25 mmHg, 95% CI -0.44 to -0.06) but there was no effect on CV (0.02%, 95% CI -0.12 to 0.16) or VIM (0.08 U, 95% CI -0.11 to 0.26). Each 1 SD increase in SD of SBP variability was associated with increased risks of hospitalization for heart failure (HR 1.19, 95% CI 1.02 to 1.38) and all-cause mortality (HR 1.12, 95% CI 1.01 to 1.25), with consistent results observed for other variability measures. No associations were observed with myocardial infarction, stroke, or kidney outcomes.

Conclusion:
Higher visit-to-visit SBP variability is independently associated with hospitalization for heart failure and all-cause mortality but not myocardial infarction, stroke, or kidney outcomes. Canagliflozin has little to no clinically meaningful effect on SBP variability.
Abstract P217: Cardiac Involvement In Creatine Deficiency Syndrome: From Transgenic Model To Humans
Authors: Del Franco Annamaria , Battini Roberta , Ferrari Chen Yu Fu , Kusmic Claudia , Forini Francesca , L'Abbate Serena , Masotti Silvia , Musetti Veronica , Borrelli Chiara , Barison Andrea , Emdin Michele , Vergaro Giuseppe
Published: September 2022

Background:
Electrocardiographic/echocardiographic abnormalities consistent with early-stage cardiomyopathy were previously reported in creatine transporter (CrT) deficiency. Nevertheless, cardiac involvement has never been accurately characterized.

Aim:
widen the current knowledge of the cardiac phenotype associated with creatine deficiency in a murine transgenic model and in patients with creatine deficiency syndrome (CDS).

Methods:
Wild type and transgenic CrT
-/y
mice (n=20) were serially evaluated
in vivo
by electrocardiogram (EKG) and echocardiography for 6 months, and
ex vivo
by histological/biochemical analyses. Nine patients with CDS (n=4 with L-arginine:glycine amidinotransferase - AGAT- deficiency, n=5 with CrT deficiency) underwent blood samples, including cardiac biomarkers, EKG, Holter monitoring, echocardiography and cardiac magnetic resonance (CMR).

Results:
Compared to wild type, CrT
-/y
mice showed a prolongation of QT interval (p=0.008) associated with a lower expression of ATPase sarcoplasmic/endoplasmic reticulum Ca
2+
transporting 2, involved in turn in intracellular Ca
2+
homeostasis. The significant reduction in mitochondrial ATP rate production (p<0.001) was attributed to a limited availability of ketone bodies, whereas ATP synthase F1 subunit alpha was not affected. No differences were detected in the left ventricular systolic function, neither by 2D-echo nor by speckle tracking. A prolonged QTc interval, detected in 7 patients at Holter monitoring, was mainly detected in case of CrT deficiency, without evidence of tachyarrhythmias. Although no impairment of cardiac function was detected at CMR, only subjects with AGAT deficiency showed an increase level of plasmatic norepinephrine and of native T1 value - 1 with an increase of extracellular volume, matching with the detection of late gadolinium enhancement.

Conclusion:
Prolonged QT interval is observed in murine model - even with a slightly impaired metabolism - and in subjects with CrT deficiency. Biohumoral signs of neurohormonal activation and an initial replacement fibrosis - the latter suggested by the increase of native T1 - are identified in subjects with AGAT deficiency, but their clinical significance remains to be determined.
Abstract P223: Hypertension Telemonitoring And Protocolized Case Management For Community-Dwelling Older Adults: A Randomized Controlled Trial
Authors: Lau Darren , Ringrose Jennifer , Finlay McAlister , Fradette Miriam , Wood Peter , Boulanger Pierre , Klarenbach Scott , Holroyd-Leduc Jayna , Alagiakrishnan Kannayiram , Rabi Doreen M , Padwal Raj
Published: September 2022

Background:
Home blood pressure (BP) telemonitoring combined with case management leads can reduce BP in adults with hypertension. However, the benefits of telemonitoring and case management for hypertension are not well established in older (age >= 65 years) adults. Interventions that can safely improve BP control in this distinct group will be critically important, particularly given recent trials showing cardiovascular benefits of intensive BP lowering in this group, and their increased vulnerability to over-treatment-related side effects.

Methods:
Twelve-month, open-label, randomized controlled trial of community-dwelling older adults comparing home BP telemonitoring (HBPM) with pharmacist-led case management versus enhanced usual care with HBPM alone. The primary outcome was the proportion achieving age-specific systolic BP targets on 24-hour ambulatory BP monitoring (ABPM). Changes in HBPM were also examined. Logistic and linear regression were used for analyses, with adjustment for baseline BP.

Results:
Subjects randomized to intervention (n = 61) and control (n = 59) groups were mostly female (77%), with mean age of 79.5 years. The adjusted odds ratio (aOR) for ABPM BP target achievement was 1.48 (95% CI 0.87-2.52, p = 0.15). At 12 months, the mean difference in BP changes between intervention and control groups was -1.6/-1.1 for ABPM (p-value 0.26 for systolic and 0.10 for diastolic BP), and -4.9/-3.1 for HBPM (p-value 0.04 for systolic BP and 0.01 for diastolic BP), favoring the intervention. Intervention group subjects had a higher rate of systolic BP < 110 (21% vs 5%, p = 0.009), but no differences in orthostatic symptoms, syncope, non-mechanical falls, or ED visits. Adherence to HBPM declined by 12 months, with 65% of trial-completers providing all 4 of the study-driven 3-monthly HBPM series.

Conclusions:
Home BP telemonitoring and pharmacist case management did not improve achievement of target range ambulatory BP, but did reduce home BP. It was not associated with major adverse consequences. Further trials will be necessary to refine HBPM interventions, improve adherence, and determine effectiveness in older individuals more conclusively.
Abstract P227: A High Fat Diet Does Not Enhance Blood Pressure Dependence On Chemerin In Normotensive Rats
Authors: Watts Stephanie W , Mullick Adam E , Garver Hannah , Orr Alexis , Fink Gregory D
Published: September 2022

Various humoral factors produced by adipose tissue are suggested to contribute to obesity associated hypertension and cardiovascular pathology. The adipokine chemerin is a frontrunner candidate. Male Dahl SS rats fed a high fat (HF; 60% kCal) diet from weaning develop severe hypertension that is profoundly reduced by weekly treatment with an antisense oligonucleotide (ASO) that disables chemerin mRNA. Here we hypothesized that feeding a HF diet from weaning to male Sprague Dawley (SD) rats would similarly increase the dependence of blood pressure (BP) regulation on chemerin. Over 17 weeks of feeding, HF fed SD rats gained significantly more weight and body fat than those fed control (10% kCal) diet. Radiotelemeters were implanted at 17 weeks of age to measure BP. Two weeks of basal BP was collected, and as we and others have shown previously, the HF diet did not increase BP in SD rats (control = 117±2.5 mm Hg; HF = 122±2.2 mm Hg). Vehicle or Gen 2.5 ASO chemerin (25 mg/kg, sc) were then given once a week for four weeks. Gen 2.5 ASO chemerin caused a slowly developing but significant reduction in BP in control rats (-14.0±2.7 mm Hg) that was not significantly different from the BP fall in HF rats (-12.4±2.3 mm Hg). RT-PCR analyses validated complete loss of chemerin mRNA in the liver and fat (primary producers of chemerin) from rats given the Gen 2.5 ASO chemerin vs control. These data show that in normal, normotensive rats a HF diet alone is insufficient to increase BP dependence on chemerin
Abstract P229: Treatment And Eradication Of
Helicobacter
Spp. In TGR(mREN2)27 Transgenic Rat Breeding Colony Does Not Alter Hypertensive Phenotype.
Authors: Graham Wayne , Cruz-Diaz Nildris , Aileru Azeez , Mitchell Erin L , Diz Debra I
Published: September 2022

The TGR(mREN2)27, transgenic homozygous (TH) rat strain is severely hypertensive, and unless treated, only 20% of males and 50% of the females survive past 20 weeks of age. Losartan-treated TH male rats are mated with normotensive female Hannover Sprague-Dawley (HnSD) rats to produce less hypertensive hemizygous (HEMI-) TGR(mREN2)27 rats with systolic blood pressure (SBP) ~180-200 mm Hg for females and males at ~20 weeks of age, allowing experimental study without antihypertensive treatment.
Helicobacter
species infection, which in rodents causes gastro-intestinal and hepatic disturbances and inflammation, compromised immunity and decreased breeding success, was detected in our TH and HnSD breeding colonies. Eradication improves animal health and breeding outcomes. However, the effects of
Helicobacter
diet treatment on phenotypic characteristics of the HEMI-TGR(mREN2)27 are unknown. TH and HnSD rats from the original WFSM colony were isolated and treated in a clean room for 8-12 weeks with Mouse Diet, Helicobacter 4 Drug Combo (Bio-Serv., Frenchtown, N.J.; pelleted diet containing 3.0 mg Amoxicillin, 0.5 mg Clarithromycin, 1.0 mg Metronidazole, 0.02 mg Omeprazole per 5 grams). Eight male and six female offspring from TH X HnSD rats and five male and six female HnSD X HnSD rats from diet-treated parents with confirmed infection eradication had tail-cuff SBP measured at 20 weeks of age for comparison with rats from the original non-treated colony. The SBPs were similar between control and treated female HEMI-TGR(mREN2)27 (191 ± 8 vs 184 ± 7 mm Hg) and between control and treated male HEMI-TGR(mREN2)27 groups (194 ± 4 vs 182 ± 3 mm Hg). Also, SBP was not different between female or male, control vs treated, HnSD rats (Female: 130 ± 6 vs 121 ± 5 mm Hg; Male:130 ± 1 vs 136 ± 5 mm Hg). Consistent with previous studies, male HEMI-TGR(mREN2)27 rats weighed significantly more than male HnSD rats (516 ± 10 vs 454 ± 16 g, p<0.01), while female mRen and female HnSD were similar (280 ± 8 vs 255 ± 4 g). Therefore,
Helicobacter
treatment and eradication in homozygous parental TGR and HnSD lines did not alter the hypertensive phenotype in HEMI-TGR(mREN2)27 rats, nor change the normotensive phenotype of HnSD rats.
Abstract P230: Hypertensive Rats Show Increased Expression Of Flavin Monooxygenases And Diminished Hypertensive Response To Trimethylamine, A Gut Bacterial Product And An Industry Air Pollutant
Authors: Konop Marek , Gawrys-Kopczynska Marta , Szudzik Mateusz , Rybka Mateusz , Mazurek Lukasz , Ufnal Marcin
Published: September 2022

Flavin-dependent monooxygenases (FMOs) are involved in a wide range of biological processes. Those enzymes catalyze oxygenation of a wide range of compounds like amines. Increasing evidence suggests a contribution of trimethylamine (TMA), a gut-bacteria product and industry pollutant, to cardiovascular disease. TMA is oxidized by FMOs to trimethylamine oxide (TMAO), an emerging marker of cardiovascular risk. We hypothesized that blood pressure phenotype may be associated with altered expression of FMOs.Gene expression and proteins of FMO1, FMO3 and FMO5, were evaluated in the kidneys, liver, lungs, small and large intestines harvested from normotensive, male, Wistar-Kyoto (WKY), spontaneously hypertensive (SHRs) and WKY with angiotensin II-induced hypertension (WKY-ANG) rats. Plasma concentrations of TMA and TMAO were measured at baseline and after the intravenous administration of TMA at dose of 45, 135 or 405μmol/kg using LC-MS. mRNA expression level of FMO1, FMO3 and FMO5 was examined. The obtained results showed that WKY, SHR and WKY-ANG rats revealed expression of FMOs in the following order of magnitude FMO3>FMO1>>FMO5.The highest expression of FMOs was found in the liver and lungs, the lowest in small intestine. In the liver and lungs, SHRs showed significantly higher expression of FMO3 than WKY (0.44 ± 0.04 vs. 0.24 ± 0.02 for liver and 0.73±0.03 vs. 0.35±0.03 a.u. for lungs respectively). It was found that, at the baseline, SHR showed significantly higher TMAO plasma concentration than WKY and WKY-ANG (10.52 ± 0.97, 3.31 ± 0.57 and 6.11 ± 0.55 μmol/L, respectively). TMA plasma level was not significantly higher in SHR than in WKY and WKY-ANG 0.15 μmol/L ± 0.01, 0.14 μmol/L ± 0.02 and 0.09 ± 0.02 μmol/L, respectively. 20 min after the infusion, SHR showed significantly higher plasma TMAO/TMA ratio than WKY and WKY-ANG, for all examined TMA doses. In conclusion, hypertensive rats show increased expression and activity of FMO3. Increased activity of FMOs may contribute to blood pressure phenotype due to altered metabolism of endogenous and exogenous amines such as TMA.
Abstract P256: Higher Angiotensin Converting Enzymes Activities Are Observed In Covid-19 Positive Volunteers.
Authors: Oliveira Lilian Caroline G , Febba Andreia C , dos Santos Lilian , Ribeiro Lys Angela F , Azinheira Nobrega Cruz Nayara , Bello Marina M , Cenedeze Marcos A , Tedesco-Silva Helio , Medina-Pestana José O , Casarini Dulce E
Published: September 2022

The high transmissibility and the broad spectrum of clinical manifestations of COVID-19 are in part due to the high affinity of SARS-CoV-2 for its receptor, Angiotensin Converting Enzyme 2 (ACE2). The depletion of the biological functions of ACE2, due to the internalization of the receptor along with SARS-CoV-2, leads to impairment of Renin Angiotensin System (RAS), which can contribute to COVID-19 pathogenesis. In addition, genetic differences in RAS may be associated with more severe symptoms and complications observed in COVID-19 patients. This study aims to perform a comparative analysis between COVID-19 positive patients and uninfected individuals, to correlate such disease profiles with ACE I/D (Insertion/Deletion) and ACE2 G8790A polymorphisms, and their enzymatic activities. The anthropometric, demographic, clinical and cardiovascular parameters of 764 individuals from Ipaussu/SP (Brazil) were evaluated. ACE and ACE2 activities were measured by fluorometric assays, and assessment of both enzymes polymorphisms was performed by PCR. In this cohort, 35,2% (269 of 764) the volunteers were positive for COVID-19. The prevalence of COVID-19 was higher among women (67%) and individuals aged between 18 and 49 years. Also, comorbidities as obesity and arterial hypertension were more frequent in the positive group, when considered individuals under 60 years old. Higher ACE and ACE2 enzymatic activities were observed in positive group (46.4 vs 41.6 and 11.3 vs 8.5, respectively). Individuals with ID genotype in the positive group presented higher ACE activity compared to individuals with same genotype in control group (46.9 vs 41.7). In the positive group, ACE activity was increased in the DD (54.5) when compared to ID (46.9) and II (37.9) genotypes. No significant differences related to ACE2 activity and polymorphism were observed. ACE/ACE2 activity ratio was higher in the COVID-19 negative group (4.7 vs 3.7). The increased ACE activity for the DD genotype was in line with the literature data for hypertension and cardiovascular diseases. We can suggest a synergic effect between ACE DD genotype and COVID-19 infection enhancing ACE activity, what may contribute to pro-inflammatory phenotype and more severe symptoms of COVID-19.
Abstract P311: Blood Pressure Control And Anti-hypertensive Drug Utilization Among Minorities In A Large Health Care System
Authors: Zoghby Ziad , Taler Sandra J , Schwartz Gary L , Garovic Vesna D
Published: September 2022

Background:
Hypertension (HTN) affects almost half of the adults in the US, and most do not have their blood pressure (BP) under control. Minorities and individuals living in rural areas experience health disparities. The goal of this study was to assess the rate of BP control among adults in a large health system that included minorities and adults living in rural areas.

Methods:
Clinical cohort study of adults with HTN who live in Southeast Minnesota and had at least one outpatient clinic encounter within the Mayo Clinic Health System in the last 12 months (5/2021-5/2022). Data was extracted through the HTN registry of the electronic health record and included age, race, ethnicity, last office BP and any active anti-hypertensives drug use.

Results:
The overall rate of HTN control among 127,644 individuals with HTN, defined as an office BP < 130/80 mmHg, was 25.7%. The rates of BP control between HTN subjects living in rural (64527, 67.3%) versus urban areas (41285, 32.3 %), were significantly lower at 24.9% vs. 27% respectively, p< 0.001. A total of 8,722 (6.8%) were non-white, including 2640 Black, 1919 Asian, and 2867 Hispanics A total of 5,734 (45%) had a SBP ≥ 140 or DBP ≥ 90 mmHg and 7178 (12.5%) did not have any anti-hypertensives in their medication list. The percentage of White and Asian persons who were not on anti-hypertensives was 12.4% and 12.5%, respectively, while a larger percentage of Black (13.8%) and Hispanic (14%) persons were not (p<0.001). The corresponding percentage was 16.7% among individuals who did not disclose their race (p<0.001).

Conclusion:
Among hypertensive adults, BP was not at goal in most patients based on their most recent office BP measurement with a higher rate among those living in rural areas. About 12.5% were not on anti-hypertensives with a disproportionate larger percentage of Black and Hispanic compared to White and Asian persons.
Abstract P313: Knockout Or Antagonism Of The Prostaglandin E2 Ep3 Receptor Is Protective In A Mouse Model Of Myocardial Infarction
Authors: Bryson Timothy D , Maxwell DruAnne , Taube David , Xu Jiang , Peterson Edward , Harding Pamela
Published: September 2022

Prostaglandin E
2
is an autacoid that acts through 4 G-protein-coupled receptors (EP1-EP4). We previously reported that expression of the EP3 receptor increases after myocardial infarction (MI) and mediates reduced cardiac function. Moreover, we reported that a selective EP3 antagonist (L798,106) prevents reduced cardiac function in Angiotensin II hypertension. We therefore hypothesized that cardiomyocyte specific knock-out and/or pharmacological inhibition of EP3 protects the heart from cardiac dysfunction after MI. To test our hypothesis, we created a conditional cardiomyocyte-specific EP3 knockout mouse (EP3 KO) by crossing EP3
fl
/
fl
mice with a tamoxifen-inducible αMHC-MerCreMer mouse. Fifteen-week-old male EP3 KO and EP3
fl
/
fl
underwent sham or MI surgery. After 2 wks cardiac function was assessed by echo and western blot was performed. There were no significant differences in cardiac function between strains after sham operation. After MI, EP3
fl/fl
mice showed significant reductions in ejection fraction (EF;68 ± 5.6 % sham vs 49.1 ± 5.2 % MI; p<0.05), coupled with increased left ventricle dimension at systole (LVDs;1.12 ± 0.08 mm sham vs 2.08 ± 0.34 mm MI; p< 0.05). However, these changes were not observed for EP3 KO mice post-MI. EF was 73.1 ± 0.68% in sham vs 69.0 ± 0.81% for MI; and LVDs was 1.09 ± 0.05 in sham vs 1.09 ± 0.04 for MI. Additionally, phosphorylated phospholamban protein levels were significantly reduced after MI in EP3
fl/fl
mice (1.14 ± 0.2 A.U. in sham to 0.42 ± 0.2 A.U. in MI; p<0.05) whereas they were not altered after MI in EP3 KO mice (1.37 ± 0.5 A.U. in sham to 1.55 ± 0.4 A.U. in MI). We next determined whether daily treatment with L798, 106 would improve cardiac function post MI. C57BL/6 mice were subject to MI and assigned to vehicle or L798,106 treatment groups (40 μg/Kg/day, s.c.) 3 days post-surgery. After 2 wks, EF was improved from 36.4 ± 4.6 % in vehicle-treated mice to 47.3 ± 5.1% in mice receiving L798,106. Fractional shortening also was improved from 17.6 ± 2.5 % in vehicle-treated mice to 24.0 ± 2.9% in L798,106 -treated mice. Altogether, these data suggest that EP3 may play a direct role in regulating cardiac function and future experiments are warranted to investigate the use of an EP3 antagonist as a potential therapeutic in heart failure.