bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 228: Targeted Treatment of SIHD: Effectiveness of Personalized Learning-Based Medical Education
Authors: Spyropoulos Jelena , Capparelli Catherine C
Published: June 2021

Study objectives:
Guideline-directed medical therapy (GDMT) to manage angina is widely underused in patients with stable ischemic heart disease (SIHD), and patients are often misdiagnosed and not managed appropriately. This study’s objective was to determine if a personalized learning-based continuing medical education (CME) intervention improved competence/performance of cardiologists and primary care physicians (PCPs) related to management of SIHD.

Methods:
An online, baseline clinical practice assessment (pre-test) consisting of 13 questions was created to assess participant’s gaps in knowledge, skills, and performance related to SIHD management. Each of the questions was mapped to one of 4 online CME activities. Incorrect responses to specific sets of questions lead to individualized recommendations to the activities which were designed to close the specific clinical gaps. Pre- and post-test responses,linked to individual learners (de-identified), were compared in each activity to identify any change in knowledge, skill, and performance. P values are shown as a measure of significance; P values <.05 are statistically significant.

Results:
Overall, 367 cardiologists and 826 PCPs participated in the assessment. 88% of cardiologists and 97% of PCPs received recommendations for 2 or more activities based on gaps in knowledge, skills, and performance. Specific improvements were observed in cardiologists’ and PCPs’ performance on management of SIHD (Table).

Conclusion:
The statistically significant improvements in competence and performance of physicians observed in this study demonstrate the benefits of using personalized educational design to address clinicians’ individual learning needs in management of patients with SIHD. This type of intervention can translate into improvements in patient care. This assessment also suggests that additional education is needed to further narrow clinical gaps regarding risk assessment, optimizing medical therapy, the use of patient-directed tools, and strategies to address adherence in patients with SIHD.
Abstract 232: Minimizing Bleeding Complications in the Cardiac Catheterization Lab
Authors: Lawson William E , Gumersell Kellie , Leippert Yvonne , Kostic Pamela , Wilbert Lisa , Schuhmacher Christine , Sproul Karen , Rowe Alison , Dervan John , Liaqat Khurram
Published: June 2021

Background:
Bleeding complications related to invasive procedures significantly increase costs, length of stay, morbidity and mortality risk. The cardiac catheterization lab [CCL] is a high risk, high volume center requiring continuous monitoring to minimize the incidence of vascular injury, hematomas/ hemorrhage, blood transfusions. Oversight extends beyond the CCL to concerns regarding post procedure access management, appropriate medication administration, ambulation, threshold for transfusion, vascular diagnostics/consultation. Based on best practices a multidisciplinary team has developed and implemented a risk stratification tool, encourages best practices, and continues to collect data to provide feedback and process improvement.

Methods:
The Stony Brook U Hospital CCL serves a Suffolk Co population of >1.5 million and provides 7x24 coverage for STEMI interventions over 912 square miles. Data post implementation of a bleeding risk stratification and optimal practices tool was collected from September of 2014 through September of 2015 to assess bleeding complication incidence [NCDR criteria], associations, and implementation of suggested practice improvements [radial access, bivalirudin, vascular closure).

Results:
A total of 1770 angioplasty procedures were performed during the monitored period [485 radial (27%), 1285 femoral (73%) approach]. In the femoral access group a total of 485 vascular closure devices (38%) were deployed. Bivalirudin [Angiomax] was used in 1,304 cases (74%); UFH was used in 466 cases (26%). Bleeding complications were identified in 69 patients during the monitored period [3.9% incidence]. Preprocedure risk screening identified 50 [72%] patients with bleeding complications as high risk and 19 [28%] patients as intermediate risk. In the patients with bleeding complications: Radial artery access was used in 16 (23%), bivalirudin in 63 (91%), vascular closure devices in 19 of appropriate femoral candidates (48%) with 12 femoral access patients (23%) receiving an Impella or IABP. Monthly case review has identified additional opportunities for improvement including: preprocedure optimization where possible, establishing transfusion thresholds, post procedure large sheathe/device management, and antithrombotic/antiplatelet medication administration.

Conclusions:
A bleeding complication task force developed a PCI bleeding risk assessment tool to identify patients at high risk of bleeding complications by NCDR criteria. Routine implementation accurately identifies high risk patients and encourages the use of best practices designed to reduce complications. Continued feedback, education and encouragement of practitioners and support staff in implementing best practices continues to improve implementation. Focused case review has provided additional opportunities for continued quality improvement.
Abstract 239: A Field Synopsis of the Role of Race in Clinical Prediction Models for Cardiovascular Disease
Authors: Paulus Jessica K , Wessler Benjamin , Lundquist Christine , Kent David M
Published: June 2021

Introduction:
Race and ethnicity-based differences and disparities have been noted in cardiovascular disease (CVD), including inequalities in access to care and differences in genetics and pharmacokinetics that affect response to treatment and development of clinical outcomes. Risk scores commonly used in CVD prevention, such as the Pooled Cohort Equation, include terms for race, yet to date there has been no systematic evaluation of the role of race and ethnicity in clinical prediction models (CPMs). To better understand the potential impact of race-specific clinical decisions encouraged by these models, we conducted a field synopsis of the role of race and ethnicity in CPMs for CVD.

Methods:
We identified CPMs in the Tufts PACE CPM Database, a systematic review of CVD-related CPMs published from 1/1990-3/2015. We report the proportion of models including the effect of race or ethnicity on CVD incidence or prognosis, summarize the directionality of the effects of race (harm or protection), and explore factors influencing the inclusion of race.

Results:
Out of 908 CPMs with CVD as either an index condition or outcome, only 24 (3%) contained a coefficient for race, ethnicity or a combination of race and ethnicity, or presented race-stratified equations. Among the overall group of CVD-related models, the racial or ethnic composition of the underlying model development cohort was reported for only 1 out of every 5 models (168/908 models (19%)), but was reported in the majority of models that included race (22/24 (92%)). Among models where the racial composition of the cohort was reported, the proportion of patients of white race was the same (79%) in models that included race vs. excluded race. The inclusion of race/ethnicity as a covariate or stratification variable was more common in models predicting incident CVD versus prognosis for patients with known CVD (9/126 (7%) versus 15/782 (2%), respectively, p<0.001)). Race/ethnicity was included infrequently even in models predicting outcomes in patient populations where racial differences have been documented: heart failure-mortality (2%, 2/107), population sample-CVD (4%, 2/52), and population sample-stroke (0%, 0/26). Among the 23 models that contained a coefficient for race/ethnicity, 11 (48%) indicated a higher risk of the outcome for patients of non-white race, while 12 (52%) indicated higher risk for patients of white race.

Conclusions:
Race/ethnicity is rarely included in CVD-related CPMs, although racial and ethnic differences in outcome risk have been documented for many conditions, and several commonly used CPMs include race. Model development from cohorts including greater numbers of non-white patients may uncover additional or more consistent racial/ethnic effects.
Abstract 240: Prevalence of Intense Systolic Blood Pressure Control in Unselected Real World Non-Diabetic Hypertensive Patients
Authors: Valadri Ravinder , Reddy Sandhya , Yang Chien-Wen , Azizi Hussain , Loughran Dan , MercadoRodriguez Estiban , Spring Deborah , Litchman Maureen , Veledar Emir , Bloom Heather , Pancholy Samir
Published: June 2021

Background:
Recent data from SPRINT trial demonstrated that, intense systolic blood pressure (ISBP) control to <120 mm Hg was associated with lower cardiovascular morbidity and mortality, compared to standard systolic BP (St-SBP) control to <140 mm Hg. We sought to determine current trend in the clinical practice in terms of intense SBP control.

Methods:
Electronic medical records from 3 different ambulatory clinics were reviewed to identify hypertensive patients with at least 3 consecutive clinic visits. Patients with diabetes, with <3 clinic visits, end-stage renal failure, and end stage liver failure were excluded. Cohort was categorized in to ISBP group and St-SBP group, when more than 50% of the clinic encounters had SBP ≤ 120 mm HG and > 120 mm Hg respectively.

Results:
1644 non diabetic patients with HTN included in the analysis. 1389 (84.5%) patients had St-SBP control whereas only 255 (15.5%) patients had ISBP control. ISBP group had significantly lower mean (±SD) SBP compared to St-SBP group; 116(±6.4) mm Hg vs. 136(±11.6) mm Hg respectively; P<0.0001. Baseline characteristics shown in table 1. ISBP group and St-SBP group had similar comorbid profile, except slightly greater prevalence of COPD in ISBP 46 (18.0%) vs 165 (11.9%); P=0.007.

Conclusion:
There is significant variation in SBP control in non-diabetic hypertensive patients in clinical practice, despite comparable comorbid profile. Furthermore, ISBP control is not a common practice in real world. Quality improvement measures should focus on promoting ISBP control in non-diabetic hypertensive patients.
Abstract 242: Influenza Seasonal Peak in Critical Congestive Heart Failure Hospitalizations
Authors: Valadri Ravinder , Veledar Emir , Deborah Spring , Litchman Maureen , Butler Javed , Pancholy Samir
Published: June 2021

Background:
Recent data from post hoc analysis of PARADIGM-HF trial demonstrated that Influenza vaccination was associated with reduced risk of all-cause mortality in patients with chronic heart failure. We sought to determine if CHF patients admitted during Influenza season (CDC definition: October to May) are critically ill compared to non-Influenza season

Methods:
Consecutive patients presented to the ER of a community teaching hospital between January 2007 to January 2016, with the diagnosis of CHF were analyzed for disposition trends. Critical illness was pre-specified as patients being admitted to either Telemetry or Intensive care unit (ICU)

Results:
A total of 3424 CHF ER-admissions were recorded in 2536 patients; 888 were readmissions. Mean age was 77 ±12 years. 1455 (57.4%) were females. 2356 (68.8%) were disposed to Telemetry, 141 (4.1%) were to ICU and 927 (27.1%) were to either medical floor or stabilized and discharged from ER. 2424 (70.8%) ER-admissions occurred in Flu season whereas only 1000 (29.2%) were in non-Flu season. There was a trend towards peak of critically ill CHF hospitalizations in Flu-season when compared with non-Flu season: 1788 (73.8%) vs 709 (70.9%); P=0.08 (Figure 1).

Conclusions:
There is a trend towards critically ill CHF hospitalizations in Flu-season. Further studies in larger data sets are needed to validate our findings.
Abstract 250: Role of First Degree Atrioventricular Block and Long Term Mortality in U.S. Veterans With Atherothrombotic Risk Factors
Authors: Patel Amish , Pollock Jonathan , Roberto Edward Sam , Aung Thein Tun , Markert Ronald , Agarwal Ajay
Published: June 2021

Background:
First-degree atrioventricular (AV) block [PR interval exceeding 200 milliseconds (ms)] on a 12-lead electrocardiogram is a common finding. Previous studies suggested that first-degree AVB has a benign prognosis, but more recent reports suggest that first-degree AVB may be associated with adverse outcomes. We investigated the relationship between PR interval and long term morality in U.S. Veterans with atherothrombotic risk factors.

Methods:
We retrospectively collected and analyzed data from a Veterans Affairs (VA) medical center for consecutive patients (October 2001 to January 2005) who presented for coronary angiography.

Results:
Of 1193 charts, 1082 had a PR interval reading recorded (mean = 172.5±30.5 ms; median = 168 ms [range 59-334]). Mean follow-up period was 103±52 months. Mean age was 63.2±10.8 years with 98% male. Mean body mass index was 30.0±5.9. The prevalence of selected comorbidities was: hypertension (88%), hyperlipidemia (79%), obstructive coronary artery disease (73%), diabetes mellitus (45%), smoker (39%), history of peripheral vascular disease (17%), and history of cerebrovascular accident (8%). Mean left ventricular ejection fraction was 47%±13%. Eighty-two percent were on beta-blockers (BB), and 25% were on calcium channel blockers (CCB) while intraventricular conduction delay was seen in 6%. In a comparison of patients with PR interval ≤200 ms (n=936) vs. >200 ms (n=146), long term mortality was higher with PR interval >200 ms (58.2% vs. 44.4%, p=0.002). Mortality rate was also higher with patients on BB or CCB vs. not on either (49.8% vs 39.7%, p=0.024). While PR interval was a significant univariate predictor of mortality, it was not significant when adjusted for the covariates listed above [not including BB and CCB] (odds ratio = 1.08 [95% Cl = 0.70 to 1.66], p = 0.73).

Conclusion:
In this study of US Veterans with atherothrombotic risk factors, the long term mortality rate was higher with first-degree AV block. PR interval has prognostic value within this specific cohort.
Abstract 001: Hypertension Induces Heightened Activity of the Adaptive Immune System
Authors: Zhao Tuantuan , Shen Xiao Z , Bernstein Ellen A , Bernstein Kenneth E
Published: June 2021

Adaptive immunity plays a key role in the pathogenesis of hypertension, but how does hypertension affect immunity? To study this, splenic dendritic cells (DC) and peritoneal macrophages were isolated from normotensive and hypertensive angiotensin II (AngII)-infused C57BL6/J mice (490 ng/kg/min, 2 wk). These antigen presenting cells (APCs) were loaded with ovalbumin (OVA) or the OVA MHC class I epitope SIINFEKL (SKL) for 3h. Then, splenocytes from OT-I mice, containing OVA-specific, CD8
+
T cells (OT-I cells), were added for 4h. Flow cytometry showed significantly more activated T cells expressing CD69 when stimulated with APCs from AngII-treated mice than equivalent cells from naïve mice (As example, DC+OVA: 2.9 ± 0.4% vs. 1.8 ± 0.3% ,P<0.05; DC+SKL: 13.9 ± 0.9 % vs. 7.4 ± 1.1%, P<0.01;). To study antigen presentation in vivo, we immunized AngII and sham treated mice with OVA and adjuvant. Consistently more OVA-specific CD8
+
T cells were induced in the blood (2.7 ± 0.2% vs. 1.2 ± 0.4%, P<0.05) and the spleen (2.6 ± 0.3% vs. 1.5 ± 0.2%, P<0.05) of hypertensive mice vs. sham when measured by flow using a H-2k
b
-SKL tetramer.

RIP-mOVA mice were also used to study if hypertension affects APC cross-presentation of self-antigens. This transgenic mouse line expresses membrane-bound OVA in pancreatic islet β cells and kidney proximal tubular cells. When OT-I cells (5x10
6
) are injected into RIP-mOVA mice, they activate by cross-presentation of OVA and cause insulitis and diabetes. RIP-mOVA mice were made hypertensive with either AngII or L-NAME. When OT-I cells were infused into hypertensive RIP-mOVA mice, they rapidly developed much higher blood glucose levels as compared to equivalently treated normotensive RIP-mOVA mice. For example, comparing mice 3 weeks after AngII and 1 wk after OT-I cells to normotensive mice 1 wk after OT-I cells, blood glucose was 331 ± 47 mg/dl in the hypertensive mice vs. 168 ± 39 mg/dl in the normotensive controls. Also, far more pancreatic islets were infiltrated by T cells in the hypertensive mice (AngII 78%, 31 of 40; L-NAME 72%, 21 of 29; control 13%, 10 of 75). In conclusion, hypertension itself is associated with higher activity of APC presentation of foreign and self-antigens which may explain why hypertension induces inflammation.
Abstract 019: Evidence that Remodeling of Insular Cortex Neurovascular Unit Contributes to Hypertension-related Sympathoexcitation
Authors: Marins Fernanda R , Iddings Jennifer A , Fontes Marco A , Filosa Jessica A
Published: June 2021

Introduction and Hypothesis:
The intermediate region of the posterior insular cortex (intermediate IC) mediates sympathoexcitatory responses to the heart and kidneys. Previous evidence indicates that hypertension alters both structure and function of neurons, blood vessels, astrocytes and microglia, disrupting the architecture of the neurovascular unit (NVU) in specific brain regions. Thus, the goal of this study is to evaluate the functional and anatomical integrity of the NVU in the intermediate IC during hypertension using
in vivo
and
in situ
experiments in male hypertensive (SHR) and normotensive (WKY) rats.

Methods:
Under urethane anesthesia, NMDA microinjection (0.2mM/100nL) was performed in the intermediate IC with simultaneous recording of renal sympathetic nerve activity (RSNA), heart rate (HR) and mean arterial pressure (MAP). NVU structure was investigated by immunofluorescence for NMDA receptors (NR1, NeuN and TOTO), blood vessels (perfused with 70kDa FITC-dextran), astrocytes (GFAP) and microglia (Iba1).

Results:
NMDA injections into intermediate IC of SHR (n=4) evoked higher amplitude responses of RSNA (Δ= WKY 26 ± 1.5 vs. SHR 44 ± 4.1 % of baseline,
P
=0.006), MAP (Δ= WKY 9 ± 1.8 vs. SHR 19 ± 2.2 mmHg,
P
=0.017) and HR (Δ=WKY 40 ± 2.5 vs. SHR 54 ± 4.9 bpm,
P
=0.044). Immunofluorescence data of the intermediate IC of SHR showed increased NMDA receptor density (WKY 16.67± 1.05% vs. SHR 24.17± 1.68%, n=6,
P
=0.003). Vascular density (WKY 1.73± 0.13% vs. SHR 2.52± 0.27%, n=10,
P
=0.015), branch and end-point number were also increased suggesting angiogenesis at the IC of SHR. Additionally, IC of SHR presented greater GFAP immunoreactivity (WKY 9.37± 2.28% vs. SHR 19.51± 3.52%, n=13,
P
=0.023) and increased contact between astrocyte processes and the vasculature (Δ= 12.8%, n=13,
P
=0.015). Skeleton analysis indicated enhanced microglia activation in IC of SHR (reduced number of branches, junctions, end-points and process length, n=13), suggesting an inflammatory process in this region.

Conclusions:
These findings suggest that the neurogenic origin of hypertension in SHR is associated with marked alterations to NVU structure within the IC contributing to enhanced NMDA-mediated sympathoexcitatory responses and maintenance of hypertension.
Abstract 021: Macrophage-dependent Impairment of the Alpha 2-adrenergic Receptor Occurs in DOCA-salt but Not Obesity-associated Hypertension
Authors: Mui Ryan , Fernandes Roxanne , Garver Hannah , Fink Gregory , Xu Hui , Galligan James
Published: June 2021

Inflammation and increased sympathetic activity contribute to hypertension. Alpha 2-adrenergic receptor (α2AR) activation decreases norepinephrine (NE) release from sympathetic nerves by inhibiting Ca
2+
channels. We hypothesized that macrophage infiltration into mesenteric arteries (MA) impairs α2AR in salt-sensitive and obesity-associated hypertensive rats. Uniphrectomized Sprague Dawley (SD) rats were given water (SHAM) or 200mg/kg DOCA (pellet, sc) and water containing 1% NaCl, 0.2% KCl (DOCA) for 4 weeks. A high fat diet (HFD, 60% kcal from fat, 0.33% NaCl, 1% K
+
) or normal fat diet (NFD, 10% kcal from fat, 0.24% NaCl, 0.36% K
+
) was given to a second group of SD rats for 20 weeks and to Dahl salt-sensitive (SS) rats for 24-26 weeks after weaning (3 weeks). Immunohistochemistry for CD163 (macrophage marker) was used to count macrophages in MA. Whole-cell patch clamp was used on dissociated celiac ganglion neurons to evaluate α2AR-mediated Ca
2+
current inhibition with NE (1 μM). Liposome-encapsulated clodronate (Clod) was used to deplete rats of macrophages. Plasma aldosterone levels were assessed by ELISA. Summary data is provided in Table 1. Systolic blood pressure was higher in DOCA vs SHAM and HFD vs NFD rats. Vascular macrophage number increased in DOCA vs SHAM but not in HFD vs NFD rats. NE inhibited Ca
2+
current to a greater degree in neurons of SHAM vs DOCA but not NFD vs HFD rats. Clodronate reduced vascular macrophages in all rats and preserved α2AR-mediated inhibition of Ca
2+
current in DOCA rats. HFD did not affect plasma aldosterone levels. Therefore, we conclude that macrophage-associated impairment of α2AR may only occur in states of mineralocorticoid and salt excess.
Abstract 029: Photoacoustic Imaging - Novel In Vivo Quantification of Placental Hypoxia in a Mouse Model of Hypertensive Pregnancy
Authors: Yamaleyeva Liliya M , Bledsoe Tiffaney , Brosnihan K. Bridget
Published: June 2021

Placental hypoxia/ischemia induces abnormal maternal and neonatal outcomes including preeclampsia and intrauterine growth restriction (IUGR). The ability to accurately determine placental oxygenation in a non-invasive way and in real-time is highly important during pregnancy as it may allow for the early diagnosis of IUGR and preeclampsia. Photoacoustic imaging (PA) is a novel preclinical and emerging clinical tool that combines optical contrast of photoacoustic laser technology with high spatial resolution of ultrasound. PA measures tissue oxygen saturation (sO2) that reflects differences in absorption spectra for oxygenated and deoxygenated hemoglobin. By using photoacoustic features of VEVO LAZR high resolution ultrasound system (VisualSonics) in a three-dimensional mode we investigated the sensitivity and accuracy of PA for placental oxygenation in C57Bl/6 mice at day 14 of gestation. Furthermore, since nitric oxide deficiency is associated with upregulation of circulatory hypoxia markers, C57Bl/6 mice were chronically treated with the nitric oxide inhibitor, L-NAME via osmotic minipumps (50 mg/kg/day; days 13 to 18 of gestation). The comparisons between scanned vs. not scanned uteroplacental units showed that PA had no effect on fetal (scanned: 0.028±0.001 vs. not scanned: 0.03±0.001 g/maternal body weight; p>0.05) or placental (scanned: 0.002±0.003 vs. not scanned: 0.002±0.002 g/maternal body weight; p>0.05) weights in C57Bl/6 mice. Changing inhaled O2 from 100- to 20% resulted on average in 12.5% reduction in total placental sO2. Systolic blood pressures were higher in L-NAME-treated vs. C57Bl/6 mice (215.8±0.8 vs. sham 99.3±4.4 mmHg; p<0.05). L-NAME infusion decreased sO2 in all areas of the placenta: labyrinth (73.6±0.97 vs. 58.6±3.4%, p<0.05), mesometrial triangle (63.0±2.0 vs. 48.9±1.0%, p<0.05), and total placenta (68.2±1.7 vs. 54.4±2.2%, p<0.05). Placental labyrinth had higher sO2 vs. mesometrial triangle area in both L-NAME infused (58.6±3.4 vs. 48.9±1.0%, p<0.05) and in C57Bl/6 (73.6±0.97 vs. 63.0±2.0%, p<0.05) mice reflecting elaborate branching morphology of the labyrinth. Our data suggest that PA imaging can detect regional differences in placental sO2 non-invasively and at different physiological states.
Abstract 033: Differential Vasopressin Receptor Expression on CD4+ T Cells from Mouse and Human Preeclamptic Pregnancies
Authors: Scroggins Sabrina M , Santillan Donna A , Peterson Jenna M , Pearson Nicole A , Sandgren Jeremy A , Devor Eric J , Gomendoza Daffney T , Pierce Gary L , Grobe Justin L , Santillan Mark K
Published: June 2021

The pathogenesis of preeclampsia (PreE) involves the failure of the maternal immune system to normally tolerate the pregnancy. Inflammatory cytokines are elevated in PreE-affected women with a concurrent decrease in anti-inflammatory cytokine production. Consistent with what other groups have observed in mouse models of hypertension during pregnancy and in human PreE-affected pregnancies, we observed increased inflammatory cytokine production and CD4+ T helper populations in our chronic infusion of vasopressin (AVP) mouse model of PreE. The mechanisms of immune modulation by AVP have not been elucidated. As increased T cell activity is involved in the development of PreE, the objective of this study was to investigate if CD4+ T cells express AVP receptors. Splenic CD4+ T cells were negatively purified from C57BL/6J saline and AVP-infused (24 ng/hour) dams. Expression of AVP receptors (AVPR) 1a, 1b, 2, and the aminopeptidase LNPEP (catalyzes AVP degradation) was determined via qPCR. Raw cycle threshold (Ct) values were normalized (ΔCt) against the 18S rRNA endogenous control. Mouse CD4+ T cells express all AVP receptors and LNPEP. By ANOVA, AVPR2 is the highest expressed receptor in CD4+ T cells from saline (N=7, p=0.002) and AVP-infused (N=10, p<0.0001) dams. Human maternal mononuclear cells, obtained from the University of Iowa Maternal-Fetal Tissue Bank (IRB #200910784) from control and PreE-affected women, were similarly analyzed. As in mouse CD4+ T cells, human control (N=27, p<0.0001) and PreE-affected (N=26, p<0.0001) CD4+ T cells most highly expressed AVPR2. AVPR1a was also highly expressed while AVPR1b was the least expressed. CD4+ T cells isolated from human PreE-affected women expressed significantly lower AVPR1a (10.0±0.3 N=27 vs. 11.1±0.2 N=0.23, p=0.009) and increased LNPEP (17.2±0.5 N=27 vs. 15.1±0.3 N=26, p=0.001) than controls. Here, we demonstrate CD4+ T cells, both mouse and human, express AVP receptors and that 1a and 2 are highest expressed. Although the actions of AVP on the vasculature are primarily mediated through AVPR1a, these data suggest AVP may differentially act through AVPR1a to mediate immune responses during PreE.
Abstract 043: Dendritic Cells Mediate Renal T Cell Activation in Hypertension
Authors: Rudemiller Nathan P , Zhang Jiandong , Hammer Gianna E , Yu Yen-Rei A , Griffiths Robert , Gunn Michael D , Crowley Steven D
Published: June 2021

Activated T lymphocytes exacerbate hypertension in part by infiltrating the kidney to promote sodium retention. Dendritic cells (DCs) are the most potent antigen presenting cells that activate T cells and have been shown to contribute to hypertension. The current studies therefore explored whether DC-mediated activation of renal T cells can exaggerate the chronic hypertensive response to angiotensin (Ang) II. First, we confirmed that renal T cells undergo DC-mediated activation during the initiation of hypertension by analyzing immune cell populations in renal tissue via flow cytometry. In Fms-like tyrosine kinase 3 ligand-deficient (FLT3L KO) mice that lack DCs, the proportions of effector (CD44
hi
CD62
lo
) T cells in the kidney were similar to wild-type (WT) controls at baseline (50±3 vs. 52±7% of CD3
+
cells). However, after 5 days of Ang II-induced hypertension, the proportions of effector T cells were dramatically higher in the WT kidney versus the FLT3L KOs (69±3 vs. 52±3% of CD3
+
cells; p<0.01), indicating that DCs activate T cells in hypertension. As DCs activate T cells in local lymph nodes, we phenotyped T cell subsets in the kidney lymph node (KLN) following 4 weeks of hypertension and detected elevated proportions of effector CD4
+
T cells compared to baseline (10.7±2.0 vs. 6.9±0.8% of CD4
+
T cells). The ubiquitin-editing protein A20 in DCs suppresses their capacity to stimulate T cells. Thus, mice with heterozygous deletion of A20 in DCs (CD11c-cre A20
flox/wt
= DC ACT) harbor spontaneously active DCs that enhance T cell activation. To test the contribution of DC-mediated T cell activation to hypertension, we measured blood pressures in WT and DC ACT mice during 4 weeks of chronic Ang II infusion (300ng/kg/min). While MAPs were similar in the 2 groups at baseline, the DC ACT mice had an exaggerated chronic hypertensive response (143±2 vs. 131±4 mmHg; p=0.04) with more severe cardiac hypertrophy (7.3±0.3 vs. 6.4±0.4 mg/gm body wt; p<0.04). The KLNs from the DC ACT animals also contained higher proportions of effector T cells than controls (12.1±0.2 vs. 8.2±0.5% of CD3
+
cells; p<0.01). In conclusion, DC-mediated activation of T cells promotes blood pressure elevation by facilitating the accumulation of effector T cells in the kidney during hypertension.
Abstract 045: Endothelial Colony Forming Cells Dysfunction Relates to Cardiovascular Alterations in Preterm Born Adults
Authors: Bertagnolli Mariane , Paquette Katryn , Sutherland Megan , Lukaszewski Marie-Amelie , He Ying , Cloutier Anik , Wu Rong , Bigras Jean-Luc , Thebaud Bernard , Luu Thuy Mai , Nuyt Anne Monique
Published: June 2021

Endothelial colony-forming cell (ECFC), a subtype of endothelial progenitor cells with high clonogenic and proliferative capacity, is present in cord and peripheral blood, participating in neovessel formation and regeneration. Cord blood ECFCs have impaired bioactivity in pregnancy complications and preterm (PT) birth. Dysfunction of cord blood ECFC is also related to complications of prematurity such as bronchopulmonary dysplasia. Although cardiovascular alterations, such as high blood pressure (BP) and left ventricular (LV) dysfunction, occur in PT subjects during adulthood, whether ECFC dysfunction beyond the neonatal period relates to such alterations is not known. OBJECTIVE: We aim in this study to assess if ECFC function relates to cardiovascular alterations in PT born adults. METHODS: Peripheral blood mononuclear cells from 30 young adults (21-28 years old) born very PT (<29 weeks gestation) and 30 at term (T, ≥37 weeks gestation) were separated by density gradient and cultured to ECFC colony formation. ECFC proliferative and angiogenic function were assessed
in vitro
by modified thymidine analogue (EdU) incorporation and tube formation in Matrigel. BP was measured by 24h monitor and LV mass index (g/cm
2
) by ultrasound imaging. All analyses were performed blind; correlations were significant when p<0.05. RESULTS: The proportion of early (<15 days) and late (≥15 days) time for ECFC colony formation was different between PT and T, with a higher frequency for late growth or no colony in the PT born group. Time to colony formation inversely correlated with ECFC proliferation rate (r=-0.57, r
2
=0.32) and tube formation (r=-0.57, r
2
=0.33) in PT, which shows ECFC dysfunction in those PT subjects with late colony formation, with no significant correlations in T. Additionally, time to colony formation has positively related to systolic BP (r=0.54, r
2
=0.29) and LV mass index (r=0.49, r
2
=0.24) in PT born subjects. CONCLUSION: Our findings demonstrate, for the first time, an association between dysfunctional circulating ECFCs and cardiovascular alterations in adults born PT. We also show that ECFC dysfunction, in PT adults, significantly relates to important cardiovascular risk factors such as high BP and increased LV mass.
Abstract 046: The Role of Tumor Necrosis Factor α and Natural Killer Cells in Uterine Artery Function During Pregnancy in the Stroke Prone Spontaneously Hypertensive Rat
Authors: Small Heather Y , Nosalski Ryszard , Morgan Hannah , Beattie Elisabeth , Guzik Tomasz , Graham Delyth , Delles Christian
Published: June 2021

Objective:
We have previously characterised the stroke prone spontaneously hypertensive rat (SHRSP) as a model of deficient uterine artery remodelling and identified an increase in pro-inflammatory TNFα relative to the normotensive WKY strain during pregnancy.

Method:
SHRSP were treated with etanercept (0.8 mg/kg) or vehicle at gestational day (GD) 0, 6, 12 and 18. Animals were sacrificed at GD18.

Results:
Etanercept reduced systolic blood pressure in the SHRSP after GD12 (ΔSBP GD 10-21 SHRSP 12.0 ± 4.17 vs. ETN 25.8 ± 4.27 mmHg; p<0.05). Uterine arteries from GD18 showed that etanercept reduced uterine artery contraction (SHRSP 57.3 ± 8.75 vs. ETN 35.2 ± 2.19 kPa; p<0.01) and increased carbachol response (SHRSP 13.8 ± 3.8 vs. ETN 40.1 ± 3.25 %; p<0.05). Uteroplacental blood flow analysed using Doppler showed that etanercept reduced uterine artery resistance index in SHRSP (SHRSP 0.79 ± 0.02 vs. ETN 0.61 ± 0.02 UARI; p<0.01). Etanercept increased litter size (SHRSP 7.80 ± 0.44 vs. ETN 12.75 ± 0.94 fetuses), reduced resorption frequency (SHRSP 66.7% vs. ETN 25.0% dams with resorption) and decreased glycogen cell loss from the placenta in SHRSP. We sought to identify the source of excess TNFα in the SHRSP. Natural killer (NK) cells (CD3-CD161+) were increased in the SHRSP relative to the WKY in the maternal circulation (WKY 1.5 ± 0.4 vs. SHRSP 6.06 ± 0.28 %; p<0.01) and placenta (WKY 11.6 ± 2.39 vs. SHRSP 659.8 ± 201.2 cells/mg; p<0.01). These NK cells produced excess TNFα in the SHRSP maternal circulation (SHRSP 6.5 ± 0.4 vs. WKY 2.5 ± 0.4 %; p<0.05) and placenta (SHRSP 65.7 ± 4.2 vs. WKY 16.9 ± 1.7 %; p<0.01) relative to the WKY. In the SHRSP placenta, etanercept treatment reduced the number of cytotoxic NK cells (SHRSP 659.8 ± 201.2 vs. ETN 148.0 ± 12.62 cells/mg; p<0.01) by down-regulating CD161 expression associated with a decrease in granzyme B production (CD161+ 71.47 ± 2.1 vs. CD161
Low
14.32 ± 0.77 % granzyme B+; p<0.01).

Conclusions:
Excess TNFα plays a causative role in adverse pregnancy outcome in the SHRSP. One source of this TNFα is an increase in NK cells during gestation in the SHRSP. Etanercept targets NK cells in the SHRSP placenta and down-regulates cytotoxic granzyme B production.
Abstract 050: mir-192-5p in the Kidney is Protective Against the Development of Hypertension
Authors: Baker Maria Angeles , Liu Pengyuan , Liu Yong , Kriegel Alison , Regner Kevin , Liang Mingyu
Published: June 2021

MicroRNAs (miRs) are short RNAs that primarily reduce protein abundance by base-paring with their target mRNA. The role of most miRs in the development of hypertension remains unknown. We performed a deep sequencing analysis of miR expression in human kidney biopsies with hypertensive nephrosclerosis or without any significant injury. miR-192-5p was one of the most abundant miRs detected and was down-regulated in hypertensive nephrosclerosis. Previous studies have shown that miR-192-5p targets the beta 1 subunit of Na/K-ATPase which drives renal tubular reabsorption. We hypothesized that miR-192-5p in the kidney protects against hypertension. We used the Dahl salt sensitive (SS) rat and a congenic rat SS.13BN26 (L26) with reduced salt sensitivity as well as Mir192 knockout mice (KO) to test this hypothesis. SS rats had a decreased level of abundance of miR-192-5p in the renal cortex compared to the L26 rats (n=9, p<0.05). The protein abundance of the beta 1 subunit of Na/K-ATPase was higher in the SS rat compared to the L26 rat (n= 3, p<0.05). Treatment with anti-miR-192-5p, delivered directly into the kidney through renal artery injection, in uninephrectomized L26 rats significantly exacerbated hypertension. Mean arterial blood pressure (MAP) of L26 rats treated with anti-miR-192-5p reached 151+/-5 mmHg at day 14 post anti-miR treatment and 4% NaCl (HS) diet, which was significantly higher than L26 rats treated with a control anti-miR and HS (135+/-5 mmHg, n=6 and 8, p<0.05). Tissues were collected in additional groups of rats at 9 days after anti-miR injection, which was just before MAP became significantly different between the two groups, for analysis of Na/K-ATPase activity. Na/K-ATPase activity was increased in the renal cortex of rats treated with anti-miR-192-5p compared to control anti-miR (9.8 +/-1.8μmole/min/μg vs 7.2+/-1.3μmole/min/μg, n=5 and 6, p<0.05). Furthermore, Mir192 KO mice treated with 1μg/Kg/min of Angiotensin II and HS for 14 days exhibited an increased MAP compared to wild-type (WT) mice (190+/-4 mmHg vs 167+/-12 mmHg; n= 3 WT and 5 KO, p<0.05). In conclusion, miR-192, particularly miR-192-5p in the kidney, confers significant protection against the development of hypertension.
Abstract 051: A Panel of CRISPR/Cas9 Genome Edited Rat Models Define Quantitative Trait Nucleotides within a Novel Rat Long Non-coding RNA Implicated in Cardiovascular Disease
Authors: Cheng Xi , Waghulde Harshal , Mell Blair , Miller Shondra , Filipiak Wanda , Saunders Thomas , Joe Bina
Published: June 2021

This study is focused on a GWAS locus for cardiovascular disease (QT-interval) on human chromosome 17. The homologous genomic segment of this human locus was previously mapped with high resolution to <42.5 kb on rat chromosome 10. The locus in rats regulates both QT-interval and blood pressure and contains a novel long non-coding RNA (lncRNA), with a large 19bp deletion/insertion polymorphism observed between the strains used to map the locus. Characterization of this novel lncRNA using rapid amplification of cDNA ends (RACE) provided evidence for the presence of more than a single isoform of the lncRNA. To further assess the role of this locus, a panel of CRISPR/Cas9 based gene-edited ‘knockout’ models of the lncRNA was developed. The lncRNA targeted rats were developed on the genomic background of the hypertensive Dahl salt-sensitive rats and harbored varied disruptions around the critical 19bp region. The rat strains with the disrupted lncRNA sequences had a significantly elevated blood pressure compared with the controls. QT-interval is currently being examined. Overall, this is the first demonstration of a CRISPR/Cas9 based targeted gene-editing approach applied to identify a novel lncRNA as a Blood Pressure Quantitative Trait Locus.
Abstract 053: RhoBTB1 is a Novel Gene Protecting Against Hypertension
Authors: Mukohda Masashi , Ibeawuchi Stella-Rita C , Hu Chunyan , Lu Ko-Ting , Davis Debbie R , Guo Deng Fu , Nair Anand R , Agbor Larry N , Wu Jing , Rahmouni Kamal , Quelle Frederick W , Sigmund Curt D
Published: June 2021

Peroxisome proliferator-activated receptor gamma (PPARγ) is a ligand activated transcription factor regulating metabolic and vascular function. We previously reported that mice (S-DN) expressing dominant-negative PPARγ in smooth muscle cells (SMC) are hypertensive, exhibit impaired vascular relaxation and enhanced contraction, and display reduced expression of a novel PPARγ target gene, RhoBTB1. We hypothesized that RhoBTB1 may play a protective role in vascular function that is disrupted in S-DN mice and in other models of hypertension. We generated double transgenic mice (termed R+) with tamoxifen-inducible, Cre-dependent expression of RhoBTB1 in SMC. R+ mice were crossed with S-DN to produce mice (S-DN/R+) in which tamoxifen-treatment (75 mg/kg, ip, 5 days) restored RhoBTB1 expression in aorta to normal. Thoracic aorta and basilar artery from S-DN showed impaired acetylcholine (ACh)-induced endothelial-dependent relaxation, which was reversed by replacement of RhoBTB1 in SMC (thoracic aorta, 43.3±4.4 vs 74.2±1.1%, p<0.01, basilar artery, 19.9±6.7 vs 48.1±12.3%, p<0.05, n=6). Aorta from S-DN mice also displayed severely decreased sodium nitroprusside (SNP)-induced endothelial-independent relaxation with a right-shifted dose-response, which was also reversed in tamoxifen-treated S-DN/R+ mice (p<0.01, n=6). Importantly, replacement of RhoBTB1 also reversed the hypertensive phenotype observed in S-DN mice (Radiotelemetry SBP, 135.9±3.9 vs 123.7±3.0 mmHg, p<0.05, n=4). To examine if overexpression of RhoBTB1 in SMC has a protective effect on other hypertensive models, Ang-II (490 ng/min/kg) was infused in tamoxifen treated R+ mice for 2 wks. RhoBTB1 expression prevented Ang-II-induced impairment of ACh relaxation in basilar artery (17.0±8.6 in control mice vs 40.7±5.3 % in R+ mice, p<0.05, n=4) and decreased SBP (166.0±7.2 in control mice vs 133.3±5.1 mmHg in R+ mice, p<0.05, n=4). We conclude that a) loss of RhoBTB1 function explains the vascular dysfunction and hypertension observed in response to interference with PPARγ in smooth muscle, and b) RhoBTB1 in SMC has an anti-hypertensive effect and facilitates vasodilatation.
Abstract 065: Overexpression of an Intracellular Angiotensin II Fusion Protein Selectively in the Mitochondria of the Proximal Tubules Elevates Blood Pressure in Mice via AT
1a
Receptor-mediated Mitochondrial Respiratory and Glycolysis Stress
Authors: Li Xiao C , Soleimani Manoocher , Nguyen Hoang , Li Hong , Zhuo Jia L
Published: June 2021

An intracrine mitochondrial renin-angiotensin system (RAS) has recently been identified in various animal and human tissues, but whether the mitochondrial RAS plays a physiological role in the regulation of blood pressure remains unknown. The present study tested whether overexpression of an intracellular angiotensin II fusion protein, ECFP/ANG II, selectively in the mitochondria of the proximal tubules alters blood pressure, and whether the effects may involve AT
1a
receptors and the Na
+
/H
+
exchanger 3 (NHE3). An adenoviral vector encoding ECFP/ANG II, a mitochondria targeting sequence, and the sglt2 promoter, Ad-sglt2-mito-ECFP/ANG II, was constructed for proximal tubule- and mitochondria-specific overexpression for 2 weeks. In adult male C57BL/6J mice, overexpression of mito-ECFP/ANG II in the mitochondria of the proximal tubules increased systolic blood pressure (SBP) significantly (Control: 116 ± 3 vs. mito-ECFP/ANG II: 128 ± 3 mmHg;
p
<0.01, n=15). The blood pressure-increasing effect of Ad-sglt2-mito-ECFP/ANG II was blocked in proximal tubule-specific AT
1a
-KO mice (Control: 105 ± 2 vs. mito-ECFP/ANG II: 104 ± 4 mmHg;
n.s
., n=7), or in proximal tubule-specific NHE3-KO mice (Control: 108 ± 3 vs. mito-ECFP/ANG II: 107 ± 3 mmHg;
n.s
., n=13), respectively. In further experiments, mouse proximal tubule cells were transfected with Ad-sglt2-mito-ECFP/ANG II for 48 h and treated with the AT
1
blocker losartan (10 μM) or the AT
2
blocker PD123319 (10 μM) to measure mitochondrial respiratory and glycolytic function using Seahorse XF Cell Mito and XF Glycolysis Stress Tests. The mito-ECFP/ANG II expression was robust and colocalized with MitoTracker® Red FM. Overexpression of mito-ECFP/ANG II markedly increased oxygen consumption rate (OCR) (Control: 139.4 ± 9.2 vs. mito-ECFP/ANG II: 236.3 ± 12.6 pmol/min;
p
<0.01, n=12) and extracellular acidification rate (ECAR) (Control: 8.8 ± 0.6 vs. mito-ECFP/ANG II: 11.8 ± 1.2 mpH/min;
p
<0.01, n=12), respectively. Losartan blocked the effects of mito-ECFP/ANG II on OCR and ECAR, whereas PD123319 had no effect. We conclude that intracellular ANG II may activate AT
1
receptors in the mitochondria of the proximal tubules to alter mitochondrial respiratory and glycolytic function and arterial blood pressure.
Abstract 071: Adam17-mediated Neurogenic Hypertension Requires Neuronal Activation of At1a Receptors
Authors: Xu Jiaxi , Sriramula Srinivas , Martin David A , Lazartigues Eric
Published: June 2021

We previously reported that neurogenic hypertension is associated with a reduction of Angiotensin Converting Enzyme 2 (ACE2) activity and an increase in A Disintegrin And Metalloprotease 17 (ADAM17) activity in the hypothalamus. Since ADAM17 is known to be expressed in multiple cell types and can be activated by various receptors, we tested the hypothesis that neuronal AT1a receptors (AT1aR) are necessary for ADAM17-mediated ACE2 shedding during neurogenic hypertension. DOCA-salt treatment (DOCA 1mg/g body weight sc + 1% saline p.o./3 weeks) was given to male neuronal AT1aR knockdown mice (AT1aR floxed crossed with Nefh-cre mice, 14-16 week-old) and their non-transgenic (NT) littermates (n=8/group). Following DOCA-salt treatment, enzyme activity assays were performed in hypothalamus of both DOCA-salt-treated and sham mice. Unlike in NT mice, ADAM17 activity was not increased by DOCA-salt treatment in the hypothalamus of neuronal AT1aR knockdown mice (153.7 ±23.8
vs.
128.5 ±11.6 FU/min,
P
=0.22). To assess whether impaired ADAM17 activation resulted from down-regulation of neuronal AT1aR, or was due to an attenuated BP rise, wild-type mice were infused with norepinephrine (NE; 4 ug/kg/min/14 d, n=6/group). Contrary to DOCA-salt hypertension, ACE2 activity in the hypothalamus remained intact during NE-induced hypertension and there was no significant increase in ADAM17 activity (59.1 ±4.3
vs.
52.3 ±11.7 FU/min,
P
=0.29). Using FACS, neurons, astrocytes and non-neuronal/non-astrocyte cells were sorted from the hypothalamus. A significant increase in ADAM17 mRNA resulting from DOCA-salt treatment (+1.72 ±0.19 fold
vs.
sham,
P
<0.05) was observed only in the neuronal population of NT mice but not in neuronal AT1aR knockdown mice. ERK phosphorylation, NOX4 expression and ROS production, all of which are involved in both intra- and extracellular mechanisms of ADAM17 activation were then assessed using Western blotting and DHE-staining. All were significantly impaired in the hypothalamus of neuronal AT1aR knock-down mice, compared to NT, after DOCA-salt treatment.

Taken together, our data provide strong evidence that activation of neuronal AT1aR is responsible for ADAM17-mediated ACE2 shedding and the maintenance of neurogenic hypertension.
Abstract 081: Obesity-related Reductions in Cardiac Baroreflex Sensitivity and Higher 24-hour Blood Pressure Variability Are Not Related to Carotid Artery and Aortic Stiffness in Young And Middle-aged Adults
Authors: Holwerda Seth , Fiedorowicz Jess , DuBose Lyndsey , Stroud Amy , Ajibewa Tiwa , Persons Jane , Woolridge Nealy , Myers Janie , Pierce Gary
Published: June 2021

Alterations in cardiac baroreflex sensitivity (BRS) and 24-hr blood pressure variability (24-hr BPV) are independent predictors of increased cardiovascular disease (CVD) risk, and occur in individuals with obesity. Obese humans are also likely to have a higher large elastic artery stiffness compared with normal-weight individuals. While an increase in stiffness of carotid and aortic arteries, the anatomical sites where baroreceptors reside, may likely be responsible in part for the decline in cardiac BRS with advancing age in adults, it remains unclear whether 1) elevated carotid and aortic stiffness are also directly associated with obesity-associated reductions in cardiac BRS in young/middle-aged individuals, and 2) if reduced BRS with obesity is associated with elevated 24hr BPV. We tested the hypothesis that lower BRS would be associated with higher carotid and aortic stiffness and 24hr BPV in young and middle-aged individuals with obesity. In a cross-sectional design, 22 normal-weight (body mass index, BMI 24.5 ± 0.6 kg/m
2
; age 35±2 yrs; 8M/14F) and 22 obese (BMI 34.2 ± 1.1 kg/m
2
; age 39 ± 2 yrs; 8M/14F) individuals underwent measures of spontaneous cardiac BRS (sequence technique), carotid artery β-stiffness (carotid tonometry and B-mode ultrasound of common carotid artery), aortic stiffness (carotid-femoral pulse wave velocity, CFPWV), and 24-hr-systolic BPV (24 hr ambulatory BP monitoring). A significant relation between cardiac BRS and 24-hr systolic BPV (r=-0.42, P<0.01) was corroborated by lower cardiac BRS (11.7±1.2 vs. 16.8±1.7 ms/mmHg, P<0.05) and higher 24-hr BPV (12.4±0.6 vs. 10.1±0.4 mmHg SD, P<0.05) among obese compared with normal-weight subjects. In contrast, carotid β-stiffness (7.8±0.6 vs. 6.9±0.4 U, P>0.05) and CFPWV (745±71 vs. 611±19 cm/s, P=0.07) were not significantly different between groups despite greater average 24-hr systolic BP in the obese vs. normal weight subjects (127±2 vs. 118±1 mmHg, P<0.05). These preliminary data suggest that an increase in carotid artery and aortic stiffness may not precede the decline in cardiac BRS and increase in 24hr BPV in young and middle-aged obese individuals, suggesting non-arterial stiffness related mechanisms for obesity-related reductions in cardiac BRS.