bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 606: Reconstitution Of Bone Marrow With WKY Cells Lowers Central/Peripheral Inflammation And Blood Pressure In The SHR
Authors: Santisteban Monica M , Zubcevic Jasenka , Kim Seungbum , Marulanda Carvajal Jessica , Zingler Michael B , Joseph Jessica , McCarter Monika A , Raizada Mohan K
Published: March 2022

Introduction:
Bone marrow (BM) is a crucial meeting point for the sympathetic innervation and hematopoietic/stem cells, including the inflammatory cells (ICs). However, the role of the BM in hypertension is not well defined. We recently demonstrated that a dysfunctional brain-bone marrow communication in rat models of neurogenic hypertension may be associated with impaired BM activity, characterized by elevated levels of ICs in the blood and BM when compared to the normotensive controls. Therefore, we hypothesized that BM of the spontaneously hypertensive rat (SHR) is pro-inflammatory, and that it contributes to hypertension in this rat model.

Methods:
6-week old female SHR rats underwent BM ablation via a lethal dose of gamma ray irradiation (950 Rads), followed by reconstitution with whole BM mononuclear cells (MNCs, 10E6 cells per rat), derived from the BM of either SHR or WKY male rats (naïve SHR n=7; SHR-SHR n=11; SHR-WKY n=9). Following a three-month recovery period, BM reconstitution was confirmed by Y-chromosome FISH in blood MNCs. Blood pressure was measured by tail-cuff plethysmography. Blood was collected for isolation of MNCs, and quantification of T-cells (CD4/CD8
+
) was carried out by flow cytometry. Brains were collected and immunohistochemistry was performed using the microglial specific marker Iba1. Microglial activation was quantified by number of microglia per 40,000um
2
in the paraventricular nucleus (PVN) of the hypothalamus.

Results:
We observed no significant difference in the mean arterial pressures (MAP) between the naïve SHR (MAP= 134 ± 5 mmHg) and the SHR-SHR groups (MAP = 137 ± 5 mmHg). However, the SHR-WKY group showed an approximate 10% decrease in MAP, to 119 ± 4 mmHg (p<0.05). This was accompanied by a 30% decrease in the circulating T-cell levels, and a 20% decrease in activated microglial cells in the PVN of the SHR-WKY when compared to the SHR-SHR group (p<0.05).

Conclusions:
These data suggest that the pro-inflammatory properties of the SHR BM contribute to hypertension by increasing both the peripheral and central inflammatory status in the SHR. These observations support the role of the BM in hypertension, especially in affecting the inflammatory status, which is a hallmark of hypertension in the SHR.
Abstract 78: Protective Effect of Nox4 in Salt-induced Hypertension in Dahl Salt-induced Rats
Authors: Cowley Allen W , Yang Chun , Kurth Theresa , Ryan Robert P , Geurts Aron M
Published: March 2022

It is recognized that oxidative stress plays a critical role in the development of salt-sensitive hypertension, especially in the kidney. The most abundant renal isoform is Nox4, but there is controversy regarding its distribution and function. There have been no studies of the role of Nox4 in a naturally occurring form of hypertension (Dahl salt-sensitive (SS) rat). We have therefore knocked out the Nox4 gene in the SS genetic background (SS
Nox4-/-
) to determine the effect upon the development of salt-sensitive hypertension and renal kidney injury in the SS rat. The SS
Nox4-/-
rat, developed using zinc finger nuclease (ZFN) techniques, exhibits an 8 base-pair frame-shift deletion within exon 7 resulting in a truncation of the Nox4 peptide by 195 amino acids with a molecular weight of 22 kD predicted (compared to the full length Nox4 peptide of 594 amino acids with a molecular weight of 68 kD). Western blot analysis confirmed the loss of the Nox4 band at ~65 kD in renal medullary tissue of SS
Nox4-/-
rats vs. SS. The functional consequence of this deletion was substantial over a 21 day period following an increase in salt intake from 0.4 (LS) to 4.0% NaCl (HS). SS
Nox4-/-

rats exhibited a change in MAP from 105±3 on LS to 130±2 mmHg (a change of 25 mmHg; n=10) on d21 of HS compared to a change from 107±2 mmHg in SS rats to 171±7 on d21 of HS (a change of 64 mmHg; n=10) as determined by telemetry. Albuminuria was virtually eliminated in SS
Nox4-/-

rats who also exhibited a large reduction of tubular cast (2.4±0.3% positive strained region vs 12.6±1.3 in SS). Glomerular injury in both the cortex and medulla was significantly less in the SS
Nox4-/-
than the SS. The % of glomeruli with an injury score of >2 (0-4 scale) was 43±1.5% (258 of 600 glomeruli) in the SS
Nox4-/-
but 65±3.9% (390 of 600) in the SS. We conclude that Nox4 plays an important role in the development of this form of salt-sensitive hypertension and renal injury in SS rats.
Abstract 90: Epoxyeicosatrienoic Acids Activation of Peroxisome Proliferator-activated Receptor Gamma Contributes to the Reduction in Glomerular Inflammation in Mice Model of Metabolic Syndrome
Authors: Elmarakby Ahmed A , Pye Chelsey , Baban Babak , Maddipati Krishna R
Published: March 2022

Studies support a link between the development of insulin resistance and the progression of renal inflammation in metabolic syndrome. Inhibition of soluble epoxide hydrolase (sEH) increases EETs levels and improves insulin signaling in type 2 diabetes, yet the molecular reno-protective mechanism (s) of EETs is/are not clearly understood. We hypothesize that decreased EETs levels mediate glomerular injury in type 2 diabetes through PPAR-γ induction. Obese db/db mice were treated with the sEH inhibitor t-AUCB (10 mg/L) alone or in combination with the PPAR-γ antagonist GW9662 (10 mg/kg ip, twice weekly) for 12 weeks. Renal EETs levels were significantly lesser in db/db mice compared to lean counterpart (100±6 vs. 162±18 ng/mg) although there was no difference in renal sEH expression between lean and obese mice. Inhibition of sEH with t-AUCB increased EETs levels in db/db mice (176±20 ng/mg). Obese db/db mice showed podocyte detachment from glomerular basement membrane and foot process effacement in glomerular tufts relative to lean and this effect was prevented with t-AUCB. Obese db/db mice also had a significantly lesser glomerular nephrin expression and higher albuminuria and nephrinuria than lean and t-AUCB treatment prevented these changes (p<0.05). Renal PPAR-γ activity and expression decreased in db/db mice together with elevation of renal p65-NFκB compared to lean mice and these changes were prevented with t-AUCB treatment. Co-administration of GW9662 with t-AUCB reversed t-AUCB’s ability to lower albuminuria and to restore glomerular nephrin expression and podocyte loss in db/db mice. Co-administration of GW9662 also blocked the ability of t-AUCB to lower T-cell infiltration, IL-17 expression and renal p-65NFκB activation in db/db mice. In conclusion, our data suggest that increased EETs levels via the inhibition of sEH in db/db mice activate PPAR-γ which in turn inhibits renal NFκB activation and reduce glomerular injury in db/db mice.
Abstract 96: Sirt-3 is Essential for Apelin Gene Therapy Mediated Improvement of Angiogenesis and Cardiac Function in Stz Mice
Authors: Hou Xuwei
Published: March 2022

Diabetic cardiomyopathy is a major diabetic complication featured by the impaired angiogenesis and cardiac dysfunction. Apelin (APLN) has been shown to prompt angiogenesis. Our recent study implicates a critical role of sirtuin 3 (SIRT-3) in the regulation of angiopoietins expression and angiogenesis. In the present study, we investigated the roles of SIRT-3 in APLN-induced angiogenesis in a diabetic cardiomyopathy model underwent ischemia. Male wild type (WT) mice and SIRT-3 knock-out (SIRT-3 KO) mice were induced into diabetic cardiomyopathy model by STZ injection and followed by left anterior descending artery ligation to induce myocardial infarction (MI). The experimental mice then underwent intra-myocardium injection with adenovirus Apelin (Ad -APLN) or β-gal adenovirus (Ad-β-gal) immediately after the ligation. Myocardial angiogenesis and cardiac function was measured 2 weeks after surgery. Our data revealed that the ejection fraction (EF) and fraction shorting (FS) were significantly improved in STZ mice received Ad-APLN treatment than in STZ mice received Ad-β-gal (EF: 64.1% ±3.2 vs 52.8%±5.6, P<0.001; FS: 34.3±2.2% vs, 26.6±3.5%, P<0.001). Ad-APLN treatment in STZ-SIRT3KO mice had little effects on EF and FS (EF: 47.9±5.2% vs, 49.7±8.2%, P=0.634; FS: 24.7±5.1% vs, 23.5±3.1%, P=0.728). The expressions of angiopoietins-1 and angiopoietins-2 were significantly increased in Ad-APLN treated STZ mice compared to Ad-β-gal treated STZ mice. These were accompanied by significant increases in myocardial arterioles and capillary densities. Ad-APLN mediated angiopoietins expressions and neovascularization were completely abolished in STZ-SIRT3 KO mice. In vitro, Ad-APLN significantly enhanced the proliferation and tube formation of endothelial cells derived from WT mice, but failed to do that in cells from SIRT 3 KO mice. Our data suggest that SIRT-3 is essential for the beneficial effect of Apelin on angiogenesis and heart function in diabetic cardiomyopathy.
Abstract 007: Effects Of Finerenone - A Novel Non-steroidal Mineralocorticoid Receptor Antagonist In A Model Of Pressure Overload-induced Cardiac Hypertrophy
Authors: Grune Jana , Benz Verena , Brix Sarah , Ban Zsofia , Salatzki Janek , Hoeft Beata , Foryst-Ludwig Anna , Klopfleisch Robert , Kolkhof Peter , Kintscher Ulrich
Published: March 2022

Activation of mineralocorticoid receptors (MR) by its agonist aldosterone induces several unwanted processes like inflammation, fibrosis, increase of blood pressure and ventricular hypertrophy. Inversely, the blockade of MR is known as a highly efficacious therapy in chronic heart failure and arterial hypertension. Therapy with currently approved MR antagonists is often limited due to side effects. Recently, new highly selective, non-steroidal aldosterone antagonists such as finerenone have been developed.

To investigate the effects of finerenone on progressive cardiac hypertrophy the transverse aortic constriction (TAC) model was used. C57BL/6 male mice underwent a TAC-operation and were treated daily by oral gavage with finerenone (fin; 10 mg/kg/d), eplerenone (200 mg/kg/d) or vehicle (veh). The treatment started one week before the TAC-operation, and was continued 4 weeks postoperative. To examine the efficacy of finerenone on myocardial wall thickening echocardiography was performed one week before and 4 weeks after TAC and left ventricular mass (LVM) was calculated. Furthermore, gene expression analysis in heart and kidney were carried out to investigate molecular mechanisms.

TAC-operated mice treated with finerenone showed a significant lower LVM relative to body weight compared with vehicle-treated mice (fin: 3.89 mg/g; veh: 4.32 mg/g; p<0.05). Also the percentage increase of LVM four weeks after TAC was significantly lower in finerenone-treated animals than in vehicle-treated mice (fin: +37.16 %, veh: +52.9 %, p<0.05). Furthermore, markers of pathological hypertrophy like atrial natriuretic factor (ANF) and β-myosin heavy chain (β-MyHC) were measured in the left ventricle, and showed a higher expression in vehicle-treated than finerenone-treated animals. Treatment with eplerenone did not significantly reduce cardiac hypertrophy after 4 weeks post TAC.

In conclusion, these data show for the first time beneficial effects of the new non-steroidal MR-antagonist finerenone on LVM development in a TAC-model. The distinct actions of finerenone when compared to eplerenone may result from different MR-blocking properties.
Abstract 011: Identification and Electrophysiological Characterization of a Novel Somatic Mutation (insT149KCNJ5) of the Potassium Channel Kir3.4 (KCNJ5)
Authors: Rossi Gian Paolo , Kuppusamy Maniselvan , Caroccia Brasilina , Bader Michael , Lenzini Livia , Stindl Julia , Bandulik Sascha , Gioco Francesca , Fishman Veniamin , Zanotti Giuseppe , Gomez-Sanchez Celso , Warth Richard
Published: March 2022

Context:
Understanding the function of the Kir3.4 (KCNJ5 gene) potassium channel through characterization of occurring novel mutations is key for dissecting the mechanism(s) of autonomous aldosterone secretion in primary aldosteronism.

Objective:
To identify novel KCNJ5 channel mutations and functionally characterize them in a large database of patients with aldosterone-producing adenomas (APA).

Methods:
We sequenced the APA and germinal DNA of 195 consecutive patients, diagnosed with the four corners criteria of the PAPY study. Among the 24.6% (48/195) APA patients who showed somatic KCNJ5 mutations we discovered a novel c.446insAAC insertion resulting in the mutant protein KCNJ5-insT149 in a patient with severe drug-resistant hypertension. The mutated cDNA generated by site-directed mutagenesis was transfected along with KCNJ3 cDNA in mammalian cells. 17α-hydroxylase, CYP11B1, and CYP11B2 were immunochemically localized in the excised adrenal gland. Whole-cell patch clamp recordings, CYP11B2 mRNA, aldosterone and intracellular Ca2+ measurement (Fura-2), and molecular modeling were performed to characterize the KCNJ5-insT149 mutation.

Results:
The patient’s high blood pressure was long-term cured; his LVMI fell from 168 to 106 g/m2 after 2 years follow-up. Compared to wild type and mock-transfected HAC15 adrenocortical cells, those expressing the mutant KCNJ5 showed increased CYP11B2 expression (expression fold change: 2.9±0.3, p<0.05 vs mock transfected cells) and aldosterone secretion (260 pg/μg RNA, p<0.05 vs mock cells). The HEK293 cells expressing the mutated KCNJ5-insT149 channel exhibited a strong Na+ inward current, and a substantial rise in intracellular Ca2+. The L-type Ca2+ channel blocker verapamil [10 μM] inhibited by 50% the pathological Na+ inward current, while both the Na+/Ca2+ exchanger blocker KB-R7943 [10 μM] and the removal of extracellular Na+ abolished it.

Conclusions:
We identified a novel mutation of the Kir3.4 channelopathy located after the pore α-helix preceding the selectivity filter, which causes pathological Na+ permeability, membrane depolarization, raised cytosolic Ca2+, and constitutive hypersecretion of aldosterone resulting in ensuing pseudo resistant hypertension.
Abstract 014: Brain Endoplasmic Reticulum Stress Mediates the Development of Non-Alcoholic Fatty Liver Disease
Authors: Horwath Julie A , Dong Frederick N , Butler Scott D , Mark Allyn L , Davisson Robin L , Young Colin N
Published: March 2022

Non-alcoholic fatty liver disease (NAFLD), characterized by an accumulation of liver lipids without excess alcohol consumption, is a major risk factor for the development of hypertension and Type-II diabetes. Liver endoplasmic reticulum (ER) stress and activation of the unfolded protein response have been implicated in the development of NAFLD. Brain ER stress has emerged as the basis of a number of chronic diseases, although the role of the central nervous system in mediating NAFLD remains unknown. We hypothesized that brain ER stress contributes to the development of NAFLD. First, male C57Bl/6 mice were instrumented with an intracerebroventricular (ICV) cannula for administration of the chemical ER stress inducer thapsigargin (TG; 1 μg/day; 3 days; n=5) or vehicle control (n=4). Daily short-term induction of ER stress in the brain with TG did not alter body weight (27±1 vs. 27±1 g; vehicle vs. TG; p>0.05), food intake or adiposity, but caused a ~40% increase in liver weight (1.04±0.03 vs. 1.39±0.10 g; vehicle vs. TG; p<0.05). ICV TG also elicited a significant increase in liver triglycerides (36±1 vs. 135±6 mmol; vehicle vs. TG; p<0.05), and real-time PCR analysis revealed upregulation of hepatic gluconeogenic and lipogenic markers in response to short-term brain ER stress (e.g.
Pepck
2.45±0.24 fold vehicle; p<0.05). Second, male C57Bl/6 mice were fed high fat diet (HFD, 60% fat, n=8), a model that causes NAFLD and brain ER stress, or remained on normal chow (5% fat, n=6) for 20 weeks. ICV cannula were then implanted for delivery of the chemical ER chaperone TUDCA (5 μg/day) or vehicle control. Inhibition of brain ER stress with ICV TUDCA over 3 days rescued HFD-induced hepatomegaly (2.2±0.2 vs. 1.5±0.1 g; HFD vehicle vs. HFD TUDCA; p<0.05) and reduced obesity-induced liver triglyceride accumulation (365±44 vs. 127±36 mmol; HFD vehicle vs. HFD TUDCA; p<0.05), independent of changes in body weight, food intake or adiposity. These data demonstrate that induction of brain ER stress caused a NAFLD phenotype, whereas inhibition of chronic HFD-induced brain ER stress reduced hepatomegaly and hepatic steatosis. Collectively, these findings illustrate a novel role for ER stress in the brain as a contributor to NAFLD.
HL63887, HL84207, AHA13POST14410020,
K99HL166776
Abstract 075: The Role of Immunological CD8+ Effector Memory T Cells In Hypertension
Authors: Itani Hana A , Wu Jing , Xiao Liang , Zhang Feng , Michell Danielle , Chen Wei , Harrison David G
Published: March 2022

Immunological memory is an important component of adaptive immunity. We have previously shown that T cells are important in hypertension. We therefore hypothesized that memory T cells contribute to long-term and repeated hypertensive challenges. To test this, mice received 2 weeks of angiotensin II, were allowed to recover for 3 weeks and then were received a low dose of ang II (140 ng/kg/min) that minimally raises blood pressure in naïve mice. This low-dose of ang II increased blood pressure to 137±6 mmHg in mice that had previously received a vehicle infusion, but to 172±12 mmHg in mice that had received ang II. In another model we treated mice with L-NAME (0.5mg/ml) in drinking water for two weeks, and then fed a high salt diet (4% NaCl) two weeks after stopping L-NAME. The high salt diet had no effect on blood pressure in naïve mice, but increased blood pressure to 150±6 mmHg in mice that had received L-NAME. In keeping with a memory T cell response, we found that either the ang II or L-NAME and high salt protocols increased CD8+ effector memory cells in the kidney by 4-fold while reducing these effector memory CD8+ T cells in the spleen by 3-fold. CD8+ memory T cells require co-stimulatory signaling between CD70 on macrophages and CD27 on T cells. We found that ang II infusion increased CD70 mRNA in isolated kidney vessels by 5 to 10-fold. Similarly, flow cytometry revealed that ang II increased macrophages expressing CD70 and CD8+ T cells expressing CD27 expression in the kidney. Preliminary results show that anti-CD70 partially blocked memory formation by reducing the hypertensive response to the second low-dose of ang II by about 20 mmHg. Thus, these studies indicate that repeated exposures to ang II promote an immune memory response of CD8+ T cells and markedly enhance the hypertensive response. The role of CD27 and CD70 signaling requires additional study but might serve as a therapeutic target.
Abstract 078: Angiotensin Converting Enzyme type 2 shedding in the central nervous system of hypertensive patients
Authors: Xia Huijing , Culicchia Frank , Moreno-Walton Lisa , Poglitsch Marko , Lazartigues Eric
Published: March 2022

ACE2 (Angiotensin Converting Enzyme type 2) is involved in the conversion of the vasoconstrictor peptide Angiotensin (Ang)-II into its vasodilatory metabolite Ang-(1-7), thereby offering a new perspective for the treatment of cardiovascular diseases associated with over-activity of the renin-angiotensin system. Our group recently reported that ACE2 shedding, a process by which ACE2 is cleaved from the plasma membrane and secreted into the surrounding milieu, contributes to the loss of the enzyme’s compensatory activity in the central nervous system (CNS) during the development of experimental hypertension. The objective of this study was to determine whether ACE2 shedding is taking place in the CNS of patients during hypertension. Unused cerebrospinal fluid, collected from patients (17 males and 27 females, aged 22-66) for diagnostic purposes, was divided in 3 groups: normotensive (n=23), hypertensive (n=9), and hypertensive controlled with medication (n=12). In addition, blood pressure values and current medications were recorded. While ELISA lacked sensitivity, soluble levels of hACE2 (sACE2) could be detected by Mass spectrometry-based RAS-Fingerprint™ in all patients. Validation experiments (n=16) indicated that pre-treatment with MLN4760 (ACE2 inhibitor) abolished the conversion of Ang-II to Ang-(1-7) by 76 ±3 %, suggesting that ACE2 is the main enzyme converting Ang-II to Ang-(1-7) in human CSF. In hypertensive patients not taking blood pressure medications, (Systolic: 157 ±10, Diastolic: 95 ±7 mmHg; n=6) CSF sACE2 activity was significantly higher (21.7 ±8.3 AU; P<0.05) than in normotensive (Systolic: 114 ±4, Diastolic: 75 ±4 mmHg; n=15) patients (6.6 ±0.9 AU). However, in patients with controlled hypertension (Systolic: 129 ±6, Diastolic: 86 ±5 mmHg; n=5), CSF sACE2 levels were normalized (6.9 ±1.9 AU). Comparison of systolic or diastolic blood pressure vs. sACE2 activity levels showed no significant correlation. These data suggest that ACE2 shedding is taking place in the CNS of hypertensive patients. Soluble ACE2 activity in the CSF is correlated with high blood pressure and could be used as a biomarker of neurogenic hypertension. AU: arbitrary unit.
Abstract 082: Oral Minocycline Reduces Blood Pressure and Restores Autonomic Balance in Chronic Ang II-Infusion Rat Model of Hypertension
Authors: Santisteban Monica M , Zubcevic Jasenka , Marulanda-Carvajal Jessica , Zingler Michael , Raizada Mohan K
Published: March 2022

Introduction:
We have established that microglial activation and dysfunctional autonomic nervous system (ANS) are associated with Ang II hypertension (HTN). The objective of this study was to test the hypothesis that oral delivery of minocycline (Mino), an anti-inflammatory antibiotic that crosses the blood brain barrier, would impact PVN microglia activation, improve autonomic function, and reverse HTN.

Methods:
SD rats irradiated and reconstituted with donor eGFP bone marrow (BM) cells were infused with saline or 200ng/kg/min Ang II S.C. for 7 wks. Additionally, rats received vehicle or Mino (50 mg/kg) by oral gavage. MAP was measured weekly by telemetry (n=3/group) and tailcuff (n=9/group). Spectral analysis of telemetry BP signal was performed for ZT12-13 at 3 wks to reveal autonomic balance.

Results:
Ang II increased MAP vs control (178±9 mmHg vs 104±2 mmHg; p<0.05), while a reduction was achieved by Mino (121±7 mmHg; p<0.05). Cardiac spontaneous baroreflex gain, measured by ΔsBRG(PI), was dampened in Ang II treated rats vs control (-0.51±0.09 ms/mmHg vs 0.21±0.23 ms/mmHg; p<0.05). This effect was attenuated by Mino treatment (-0.08±0.08 ms/mmHg; p<0.05). Vasomotor sympathetic tone (ΔLF[SBP]) and vasovagal balance (ΔLF[SBP]:HF[PI]) were increased in Ang II rats (1.0 ±0.48 mmHg2; 0.12±0.06 mmHg2/ms2, respectively; p<0.05), and were reversed by Mino (-0.83±0.45 mmHg2; -0.11±0.05 mmHg2/ms2, respectively; p<0.05). In contrast, no significant changes were observed in cardiac parasympathetic drive (ΔHF[PI]) and cardiac sympathetic tone (ΔLF[PI]:HF[PI]). Immunohistochemical analysis revealed 5.7 fold increase (p<0.05) in GFP+/Iba1+ cells in PVN of Ang II group vs control, indicating an increase in BM-derived microglia/macrophages. Treatment with Mino was associated with a 32% decrease in these cells. Blood CD4+8+ cells, indicative of peripheral inflammation, increased by ~29% in the Ang II group, and decreased in the Ang II/Mino group by 31% (p<0.05).

Conclusions:
These observations demonstrate that oral Mino treatment inhibits microglia activation, attenuates autonomic dysfuction and peripheral inflammation that arrests the progression of HTN. This suggests that Mino might be an effective therapeutic agent to target neurogenic HTN.
Abstract 206: Incorporation Of Clinical Parameters To Morisky Medication Adherence Scale Assessment Improves Its Ability To Predict Non-adherence In Patients With Resistant Hypertension
Authors: Pandey Ambarish , Raza Fayez , Velasco Alejandro , Brinker Stephanie , Ayers Colby , Arbique Debbie , Price Angela , Vongpatanasin Wanpen
Published: March 2022

Background:
Morisky Medication Adherence scale (MMAS), a self-report questionnaire for screening non-adherence, has been validated in patients with uncomplicated hypertension. However, its accuracy in predicting non-adherence in patients with resistant hypertension (RH) is not known.

Methods:
A retrospective chart review was performed in 91 patients with RH referred to hypertension clinic from 2009 to 2013 that filled out a MMAS survey during the initial clinic visit. Non-adherence was confirmed by presence of undetected serum levels of at least 1 prescribed antihypertensive drug by therapeutic drug monitoring (TDM).

Results:
By MMAS, 27% (25/91) patients had low adherence, 30% (27/91) medium adherence, and 43% (39/91) high adherence with no significant difference in mean BP among these groups. MMAS was limited in its ability to predict non-adherence when validated against TDM (sensitivity=40%; specificity = 33%) and was not an independent predictor of non-adherence. However, clinical characteristics such as Heart rate (HR), Heart Failure (HF) and systolic BP (SBP) were independent predictors of non-adherence after multivariable adjusted logistic regression analysis. Area under the curve (AUC) resulting from receiver operator curve (ROC) analyses was greater for clinical predictors (SBP+HF+HR) than the MMAS score alone (0.89 vs. 0.65). When SBP, HF, and HR were added to model with MMAS score, the AUC increased significantly (see Fig; MMAS= 0.65; MMAS + SBP +HF + HR= 0.91).

Conclusion:
Ability of MMAS to predict non-adherence in patients with RH can be significantly improved by incorporating clinical parameters readily available for physicians in clinical practice.
Abstract 207: Angiotensin -II (Ang-II) Induces Endoplasmic Reticulum (ER) Stress-mediated Overproduction of Mitochondrial Reactive Oxygen Species (mROS) and Alterations in Mitochondria-associated Membrane (MAM) in Neural Cells
Authors: Sarkar Pallabi , Patel Unisha , Wang Gang , Iadecola Costantino , Mark Allyn L , Davisson Robin L
Published: March 2022

ER stress and mROS are identified as key mediators of Ang-II-induced neurogenic hypertension. Although ER and mitochondria are linked both structurally and functionally, the interplay between these two organelles in neural cells and in the subfornical organ (SFO), a key forebrain region in Ang-II hypertension, is unknown. Previously we reported that Ang-II causes mitochondrial fragmentation in murine SFO neurons. Here we tested the hypothesis that Ang II-induced mitochondrial dysfunction is mediated by ER stress and disruption of the MAM. Neuro-2A cells were incubated with Ang-II (300 nM, 24 h) and real-time PCR was performed to examine expression of key molecular markers of MAM (n=3-5). Dynamin related peptide 1 (drp1), a MAM marker and mitochondrial fission protein, was increased in the presence of Ang-II (1.77±0.17-fold vehicle, p<0.05).This effect was blocked when cells were co-treated with the chemical ER stress inhibitor TUDCA (0.77 ± 0.27 fold vs. Ang-II; p<0.05). Ang-II also increased the expression of phosphofurin acidic cluster sorting protein 2 (pacs2, another MAM marker) and the co-administration of TUDCA was able to block this effect (Ang-II: 1.5 ± 0.02-fold vs. vehicle; TUDCA+Ang-II: 0.9 ± 0.03 fold vs. Ang-II; p<0.05). Ang-II-induced mROS production, measured by MitoSox Red dye (n=11-15), was decreased in the presence of TUDCA (12 ± 4% increase from vehicle compared to a 58 ± 2% increase with Ang-II alone ; p<0.05). BD-1047, a chemical disruptor of MAM, also prevented the Ang-II-induced increase in MitoSox fluorescence (9.6 ± 0.2 % vs. Ang-II alone; p<0.05). Furthermore, when MDivi-1, a selective drp-1 blocker, was added, a similar blockade of mROS production was observed (11.9 ± 0.8 %; p < 0.05) In dissociated SFO cells from adult male C57Bl/6 mice (n=8-23), Ang-II-induced mROS production (12.9 ± 2% increase from vehicle, p< 0.05) was blocked by TUDCA (0.8 ± 0.01%, p<0.05) and by losartan (2.2 ± 1.6%, p<0.05). These data demonstrate that induction of ER stress by Ang-II leads to mitochondrial fragmentation mediated by MAM disruption, which then results in mROS overproduction. Collectively, our data suggest a novel role for ER-MAM interaction in Ang-II-mediated hypertension.
Abstract 211: Increased Blood Pressure Regulation By Nitric Oxide In Obese African American Women Unmasked By Autonomic Blockade
Authors: Shibao Cyndya , Gamboa Alfredo , Celedonio Jorge E , Arnold Amy C , Okamoto Luis E , Diedrich Andre' , Biaggioni Italo
Published: March 2022

African American women have the highest prevalence of hypertension (46%) in the world. African Americans (AA) have decreased vascular reactivity to pharmacologic (methacholine, nitroprusside, isoproterenol) and physiological (ischemia) stimuli. Such findings could be explained by racial differences in nitric oxide (NO) release, NO responsiveness, or by a differential suppression of NO by the sympathetic nervous system. The goal of this study was to evaluate tonic restraint of blood pressure by NO during complete autonomic blockade in obese AA vs. white women. We hypothesized that if AA women had a primary NO “deficiency” then they would have less increase in systolic blood pressure (SBP) during intravenous infusion of the NO synthase inhibitor N(ω)-monomethyl-l-arginine (L-NMMA, 250 ug/kg/min) during autonomic blockade. We studied 8 obese AA women and 9 obese white women matched by age (41±6.4 vs. 42±8.2 years, P=0.66), BMI (34±4.4 vs 35±4.1 kg/m2, P=0.83) and seated SBP (121±11 vs. 125±7 mm Hg, P=0.49). Antihypertensive medications were discontinued for 2 weeks and smokers were excluded. All subjects underwent complete autonomic blockade with the ganglionic blocker trimethaphan (TMT, 4 mg/min) followed by L-NMMA. The alpha-1 adrenergic agonist phenylephrine was titrated to clamp systolic blood pressure at 110 mm Hg prior to infusion of L-NMMA. SBP response to L-NMMA was greater in obese AA women (54±13.6 vs. 39±9.6 mm Hg in whites, P=0.02, Figure) suggesting that NO, in the absence of autonomic influences, is an important regulator of blood pressure in obese AA women. This suggests that sympathetic activation contributes to "NO deficiency" in AA.
Abstract 234: AGEs Enhance the Metabolism of Angiotensin-(1-7) and Lower Intracellular Peptide Expression
Authors: Alzayadneh Ebaa M , Wilson Bryan A , Marshal Allyson C , Chappell Mark C
Published: March 2022

Advancedglycated end products (AGEs) including glucose- and methylglyoxal-modified albumin (MGA) may be important mediators of diabetic pathologies. An accompanying abstract described AGE-dependent cellular hypertrophy and myofibroblast transition (MT) in NRK-52E renal epithelial cells that was abolished by treatment with the alternative angiotensin (Ang) product Ang-(1-7); the inhibitory effect of Ang-(1-7) was likely mediated by attenuation of ERK and TGF-β stimulated pathways. AGEs evoke multiple signaling events, but the impact on the Ang-(1-7) system is unknown. The present study addressed whether AGEs suppress the Ang-(1-7) axis, and thus facilitate their deleterious actions. MGA exposure for 48 hrs significantly reduced the cellular expression of Ang-(1-7) [396 ± 59 vs. 219 ± 54 fmol/mg protein; p<0.05] in the NRK-52E cells; however, the cellular content of Ang II and Ang I was not changed. We then assessed the processing pathways of Ang I to Ang-(1-7) in the 100,000 xg soluble fraction, as well as the activity secreted extracellularly in the media. Conversion of Ang I to Ang-(1-7) was also not changed following MGA exposure in the cellular supernatant [56 ± 7 vs. 66 ± 8 fmol/min/mg protein]. In contrast, the metabolism of Ang-(1-7) to Ang-(1-4) was significantly enhanced in the soluble fraction of the MGA exposed cells [175 ± 9 vs. 115 ± 11 fmol/min/mg protein, p<0.05]. The Ang-(1-7) endopeptidase activity was abolished by the thiol inhibitor PCMB, but was insensitive to the metallopeptidase inhibitor JMV390. AGE slightly but significantly induced the metabolism of Ang-(1-7) to Ang-(1-4) in the cell media [1.36 ± 0.01 vs. 1.42 ± 0.01 fmol/min/ml, p<0.05]. In this case, the secreted endopeptidase exhibited marked sensitivity to JMV390 [IC50 = 7.6 nM]. Finally, chronic treatment with the ERK1/2 inhibitor PD98059 abrogated the increase in Ang-(1-4), and suggests that downregulation of the peptidase is linked to AGE-induced MT. We conclude that the cellular actions of AGE may reflect a loss of Ang-(1-7) tone through enhanced metabolism of the peptide. Moreover, we identify two distinct endopeptidase activities that metabolize Ang-(1-7) to Ang-(1-4) and both are increased following AGE exposure.
Abstract 239: Natural Language Processing Identifies Important Side-effects In Very Elderly Patients With Hypertension,
Authors: Odunukan Olufunso W , Komadur Elayavilli Ravikumar , Cha Stephen , Liu Hongfang , Rahman Parvez A , Roellinger Daniel , Nyman Mark A
Published: March 2022

Background:
Concerns for side-effects makes the identification of treatment targets of hypertension in the very elderly challenging. We sought to utilize natural language processing (NLP) to evaluate the prevalence of prespecified side-effects in a cohort of very elderly (≥80 years) patients.

Methods and Results:
We conducted a two-year, cross-sectional study involving 5169 very elderly patients with and without hypertension seen by primary or specialist providers. Hypertension patients had to have a prior hypertension ICD-9 code in the electronic medical records problem list and ≥2 office visits in 2008 and 2009. We used NLP to analyze extracted free text reports from last clinic visits between 2008 and 2009. Of the 10 prespecified side-effects, hypotension, fatigue, and dizziness were the commonest three conditions and hypotension was significantly higher in the HTN patients. Of the 2835 patients with no prior hypertension history and 2334 hypertension patients identified, the prevalence of each prespecified item in both groups were: acute renal failure (0.3% v 0.6%, p=0.13); syncope (2.8% v 2.2%, p = 0.17), dizziness (3.1% v 4.0%, p=0.09), history of fall (1.0% v 0.9%, p=0.65), fatigue (5.4% v 4.5%, p=0.19), femoral neck fracture (0.18% v 0.04%, p=0.16), palpitations (1.2% v 1.5%, p=0.35), hypotension (4.1% v 6.6%, p<0.001), hyperkalemia (1.1% v 1.7%, p=0.04), hypokalemia (0.4% v 0.9%, p=0.01). At least one presumed side-effect was present in 17.5% of all 5169 patients. Among hypertension patients, there was no significant difference in the prevalence of the side-effects among patients with BP above (578) or below 140/90 (1756) except for acute renal failure (1.21% vs 0.40%, p = 0.03).

Conclusion:
Analysis of free-text outpatient notes using NLP showed that hypotension and potassium abnormalities were significantly more prevalent in very elderly patients with hypertension compared to similarly aged patients with no hypertension history.
Abstract 241: Burden of hypertension on hospitals in USA: Analysis of Nationwide Emergency Department Sample Data
Authors: Aggarwal Sourabh , Agrawal Yashwant , Gupta Vishal
Published: March 2022

Introduction:
Cardiovascular diseases are a leading cause of morbidity and mortality in USA. Hypertension (HTN) is a major risk factors for cardiovascular disease with unclear extent of burden on USA healthcare. This study was done to identify the burden imposed by HTN independently on hospitals in USA.

Methods:
We used Nationwide Emergency Department (ED) Sample data to extract data for patients who visited ED with first listed diagnosis of HTN. Clinical classification software code 98 and 99 were used to identify patients with essential HTN and HTN with complications and secondary HTN respectively. Data was extracted for the years 2006 to 2011.

Results:
We identified a total of 3,938,456 ED visits with first listed diagnosis of essential HTN for years 2006-11 with admission rate of 9.76% (384,495 out of 3,938,456). The rate of ED visits increased from 190.1 visits per 100,000 persons in 2006 to 238.5 visits per 100,000 persons in 2011 (p<0.01). However, the admission rates decreased significantly from 10.47% (59,374 out of 567,232) in 2006 to 8.85% (65,729 out of 742,997) in 2011 (p<0.01).

We identified 1,429,308 ED visits with first listed diagnosis of HTN with complication and secondary HTN for years 2006-11 with admission rate of 73.41% (1,049,199 out of 1,429,308). The rate of patient visits to ED increased from 71.2 per 100,000 persons in 2006 to 84.7 per 100,000 persons in 2011 (p<0.01), however the admission rate decreased significantly from 77.79% (165,254 out of 212,440) in 2006 to 68.75% (181,527 out of 264,050) in 2011 (p<0.01). The in-hospital mortality for admitted patients decreased from 1.95% in 2006 to 1.25% in 2011 (p<0.05).

Conclusions:
Our study revealed that number of patients visiting ED with first listed diagnosis of essential HTN and HTN with complication and secondary HTN increased steadily from 2006 to 2011. Though admission rate for patients visiting ED decreased, the aggregate number of admitted patients increased leading to increased burden on both ED and in-patient healthcare facility indicating still large unmet need for better outpatient care to prevent these ED visits and hospitalizations. The decreasing in-hospital mortality rate points to improving care of hospitalized patients.
Abstract 260: Toll-like Receptor 3 (TLR3) Activation Causes Contraction Of Vascular Smooth Muscle Through ERK1/2 Pathway And Increases Blood Pressure In Mice
Authors: Carrillo-Sepulveda Maria Alicia , Hardigan Trevor , Spitler Kathryn , Webb R. Clinton
Published: March 2022

Emerging evidence has suggested that viral infection can trigger hypertension. TLR3, a pattern-recognition receptor of the innate immunity, is activated by double-stranded RNA (dsRNA) from viruses or necrotic tissue. Vascular smooth muscle cells express TLR3; thus we hypothesized that TLR3 activation causes an increase in ERK1/2 signaling-induced vascular contractility and subsequently increases blood pressure. Stimulation of mouse C57bl/6 aortic smooth muscle cells (ASMC) with polyinosine polycytidylic acid (Poly I:C) (10 μg/mL, 60 min), a synthetic analog of dsRNA, resulted in increased myosin light chain phosphorylation (pMLC) (2.2 fold vs. unstimulated cells, p<0.05), a marker of vascular contraction. This effect was accompanied by increased phosphorylation of caldesmon (pCaD) on Ser789 (3.35 vs. unstimulated cells, p<0.05) as well as activation of ERK1/2 (2.56 fold increase vs. unstimulated cells, p<0.05). Pre-treatment with PD98059 (50μM, 30 min), an ERK1/2 inhibitor, decreased Poly I:C-mediated phosphorylation of CaD and MLC. Treatment with Poly I:C for 24 hours downregulated TLR3 expression by 40% in ASMC (p<0.05). Vascular studies performed on a tension myograph showed that aortas from mice treated for 2 weeks with intraperitoneal injection of Poly I:C (20mg/Kg every 48 hours) exhibited increased contraction to norepinephrine compared to control mice (EMax as %of maximum KCl response: 126± 4.56% vs. 108± 1.61%, respectively). This effect was attenuated in aortas from Poly I:C mice pre-incubated with 10uM PD98059 (EMax as %of maximum KCl response: 110.6± 5.12%). Systolic blood pressure (mmHg) was higher in Poly IC mice compared with control (128 ± 2 vs 159 ± 2). Increased spleen and kidney weight (mg) were observed in Poly I:C mice (77.63 ± 4.67 vs. 127.06 ± 7.70 control and 141.10 ± 2.42 vs. 180.35 ± 8.85 control, respectively).Our results show that activation of TLR3 in VSMC causes ERK1/2-mediated phosphorylation of MLC and CaD, thereby contributing to augmented contraction and development of vascular dysfunction and hypertension. Our data provide the functional assessment of the role of TLR3 in vascular contractile events, suggesting TLR3 as a potential new therapeutic target in vascular dysfunction and regulation of blood pressure.
Abstract 269: Look Ma, No Hands! Cardiopulmonary Resuscitation
Authors: Brown Coffee , Fullerton Lynne , Mayschak Trevor , Edwards Isaac , Light Kelli , Watkins Matthew
Published: March 2022

Background:
CPR saves lives. Community-based, bystander CPR increases survivability by reducing time to compressions. Inadequate chest compressions have been associated with poor patient outcome. There is substantial inter-operator variability in quality and duration. While training laypersons in CPR, we observed that individuals lacking upper body strength had difficulty sustaining effective compressions. An improvised compression technique, transferring the force to the operator’s forearm, immediately resolved this issue.

Objective/hypothesis:
We hypothesized that a modified technique would be associated with increased duration of adequate compressions relative to standard CPR.

Methods:
Using a convenience sample of 18 paramedic students, we standardized this forearm technique and compared duration of adequate compressions with Standard CPR using Laerdal PC SkillReporting software. Trials were terminated upon 5 consecutive inadequate compressions. Evaluators were blinded to which sequence-randomized technique was being used. Our case-crossover study was approved by the university IRB.

Results:

Limitations:
The small, convenience sample of young, fit providers trained in CPR was not representative of the group for whom the improvisation was originally devised. Larger scale testing with greater diversity is indicated.

Conclusions:
Our pilot study supported our hypothesis that the forearm CPR method was not inferior to the standard method with respect to time to fatigue. Having the mannequin at hospital bed height likely influenced the effectiveness of both techniques, so ensuing studies will use a mannequin on the ground.
Abstract 272: Divergent Neural Control of Blood Pressure vs Resting Metabolic Rate by Angiotensin: The Role of AT
1A
Signaling in Leptin-Sensitive Cells
Authors: Claflin Kristin E , Stauss Harald M , Grobe Justin L
Published: March 2022

The renin-angiotensin system (RAS) and leptin both contribute to resting metabolic rate (RMR) and blood pressure control, and thus may contribute to obesity-hypertension. Previously we demonstrated a required role for angiotensin AT
1A
receptors in the brain in RMR, and renal and sympathetic nerve activity (SNA) responses to leptin. Here we tested the hypothesis that AT
1A
receptors, specifically localized to leptin-sensitive cells, are required for the blood pressure and RMR effects of brain ANG. The AT
1A
gene was conditionally disrupted in cells that express the long (b) form of the leptin receptor (ObRb-Cre x AT
1A
flox
mice; “KO”), and blood pressure was assessed by radiotelemetry under baseline and deoxycorticosterone (DOCA)-salt stimulated conditions. While 24-hr blood pressure (MAP; control n=10, 115 ± 4, vs KO n=9, 115 ± 6 mmHg) and heart rate (control n=10, 561 ± 9, vs KO 563 ± 11 BPM) were not different between control and KO mice, baroreflex sensitivity was significantly increased in KO mice at baseline (control n=6, 3.4 ± 0.7, vs KO n=6, 5.6 ± 0.5 msec/mmHg, P<0.05). DOCA-salt significantly increased MAP (control=8, 130 ± 4, vs KO n=7, 129 ± 3) and decreased baroreflex sensitivity (control=6, 3.0 ± 0.5, vs KO n=5, 2.8 ± 0.5, P<0.05 vs baseline) in all mice. In contrast, RMR responses to DOCA-salt were abolished in KO mice (baseline n=2, 0.2 ± 0.01, vs DOCA-salt n=2, 0.2 ± 0.02). We conclude that AT
1A
signaling specifically in leptin-sensitive cells is critically involved in RMR control, but is relatively dispensable for cardiovascular control. To further characterize the cells that mediate the leptin-AT
1A
interaction critical for RMR control, we performed immunohistochemical staining for adrenocorticotropin (ACTH) in the arcuate nucleus of mice that express GFP via the AT
1A
promoter (“NZ44”). Complete lack of colocalization supports the conclusion that AT
1A
are not expressed by proopiomelanocortin (POMC) neurons. In contrast, KO mice exhibit a baseline increase in glutamate decarboxylase mRNA in the arcuate nucleus, consistent with a possible localization of AT
1A
upon agouti related peptide (AgRP) neurons. AT
1A
may therefore suppress AgRP neuron γ-aminobutyric acid production and thus disinhibit POMC neuronal activity, to ultimately stimulate RMR.
Abstract 273: Aerobic, Resistance And Combined Exercise Training Across The Lifespan Are Effective In Improving Heart Rate And Blood Pressure Variability In Obese Mice
Authors: Sartori Michelle , Santos Fernando , da Silva Maikon B , De Angelis Katia , Irigoyen Maria C
Published: March 2022

The aim of this study was to compare the effects of aerobic, resistance and combined exercise training on autonomic parameters in youth obese mice. Female mice (4 weeks) were randomized into: wild type sedentary (WS, n=11), ob/ob sedentary (OS, n=11), ob/ob aerobic trained (OA, n=8) (treadmill,5days/w, 60min/d, during 8 wks), ob/ob resistance trained (OR, n=8) (ladder climb, 5 days/w, during 8 wks) and ob/ob combined trained (OC, n=8) (alternate days of aerobic and resistance exercise) groups. Blood pressure (BP) signals were directly recorded using a data acquisition system. OS group presented higher blood glucose levels (169±8mg/dL) than WS animals (121±6mg/dL). In addition ob/ob trained groups (OA, OR and OC) groups had reduced blood glucose (OA:133±3, OR:155±9 and OC:143±11mg/dL) in comparison with OS group. Systolic, diastolic and mean BP and heart rate were similar among groups. The ob/ob mice showed an decrease in pulse interval variance and RMSSD compared with WS group. The three types of training were effective in increasing these parameters in trained mice compared with ob/ob sedentary mice. The BP variability was similar among groups; however, the low frequency band of BP was higher in the sedentary groups (WS:10±1 and OS:10±1mmHg
2
) compared to OA (5±1mmHg
2
) and OR (5±1mmHg
2
) groups. . The spontaneous baroreflex sensitivity was lower in ob/ob groups compared to WS (3±0.3 ms/mmHg). Furthermore, the OS group showed decrease in this parameter (0.6±0.3 ms/mmHg) compared to OA (1.7±0.2ms/mmHg) and OR groups (2±0.6ms/mmHg). The OA, OR and OC groups were similar in relation to low frequency band of BP and baroreflex sensitivity. Thus, our results suggest that physical training across the lifespan, either aerobic and/or resistance exercise, can be effective in attenuating the autonomic dysfunctions associated with obesity in mice.
Abstract 284: Lower Nrf2 Activity And Decreased Levels Of Pdi And No In The Kidney Contributes To Hypertension In The Aging Fbns
Authors: Wang Xitao , Asghar Mohammad
Published: March 2022

We recently reported that age-associated oxidative stress is causal to higher renal angiotensin AT1 receptor (AT1R) function and blood pressure (BP) in the aging Fischer Brown Norway (FBN) rats. This suggests that redox homeostasis is critical for normal AT1R function and blood pressure (BP). Our objective was to understand the mechanism responsible for the higher oxidative stress in the aging kidneys contributing to impaired AT1R function and high BP in the aged FBNs. Kidneys from adult (3-month) and aged (21-month) FBN rats were isolated. Nrf2 transcription factor, involved in transcriptional activity of diverse antioxidant enzymes genes, was determined in the nuclear and cytosolic fractions of cortical tissues by Western blotting. Nuclear accumulation of Nrf2 is an index of its activation. Protein disulfide isomerase (PDI), an enzyme involved in isomerization reaction, was determined in the cortical homogenates. Moreover, nitrate/nitrite levels were determined in the urine as an index of NO levels in the kidney. We found that the levels of Nrf2 were decreased (adult vs old: 1.86±0.69 vs 0.87±0.18, -53.2%) in the nuclei but increased (adult vs old: 1.20±0.27 vs 1.51±0.36, +25.8%) in the cytosol of aging kidneys. PDI levels also were decreased (adult vs old: 5.38±2.60 vs 3.59±1.76, -33.3%) in the aging kidneys. Furthermore, nitrate/nitrite levels were lower (adult vs old: 372±73 vs 211±30, -43.3%) in the urine from aged than in adult rats. These data suggest that impaired Nrf2 activity in the aging kidneys contributes to increased oxidative stress, renal AT1R function and BP in the aged FNBs. Furthermore, these data also indicate that PDI in an NO-dependent pathway regulates Nrf2 activity contributing to redox-balance in the kidney.