bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 628: The Therapeutic Effect Of Indian Classical Music on Physiological And Psychological Parameters Of Hypertension in Geriatric Patients
Authors: Godbole Manasi M , Basavaraj Anita
Published: March 2022

Background:
Old age is associated with cardiovascular problems including isolated systolic hypertension and uncontrolled hypertension. Poly pharmacotherapy especially in geriatric patients can cause side effects due to drug interactions and there is a need for less pill burden and adjuvant therapy. The basis of calming effect of Indian classical music finds its mention even in mythological anecdotes. Moreover, music therapy is safe, inexpensive and non-invasive.

Aim:
To evaluate the effect of Indian classical music as evidence based treatment for hypertension and anxiety level reduction.

Method:
Pre-selected 60 geriatric patients (60-85years) were randomly assigned to Study Group(SG){n=30}and Control Group(CG){n=30}.The SG, in addition to conventional treatment, received 20 minutes of pre-recorded Indian classical music therapy for 4 sessions. The CG received standard treatment alone. Blood pressure (BP), pulse rate (PR) and respiratory rate (RR) were taken before and after each session. Psychological assessment was done at the start and at the conclusion of final session.

Results:
Comparing pre and post parameters of all the sessions, the intervention resulted in a statistically significant reduction in BP and RR in both groups and significant reduction in PR only in the SG. Further analysis revealed a more statistically significant reduction in BP, PR and RR and anxiety levels in the SG as compared to CG after music session.

Conclusion:
This study indicates the use of Indian classical music as an innovative therapeutic approach in modern era for prevention or in conjunction with antihypertensive medications for blood pressure management in geriatric patients.
Abstract 629: Gender Differences In The Expression Of Cardioprotective Myeloid Cell Leukemia 1 (Mcl-1) In Diabetic Male And Female Rats
Authors: Lum-Naihe Kelly , Slusarz Anna , Luck Christian , Arnold Nicholas , Koppula Purushotham R , Pulakat Lakshmi
Published: March 2022

Diabetes Mellitus (DM) is an important risk factor for cardiovascular disease (CVD) and young, diabetic women are more susceptible to CVD compared to age-matched men and non-diabetic women. It is unclear how the progression of DM differentially affects cardiac pathology in males and females. In this context, recent studies have shown that myeloid cell leukemia 1 (MCL-1) plays an important role in cardiac protection and cardiac-specific ablation of Mcl-1 causes fatal dilated cardiomyopathy. We hypothesized that there is a gender difference in the expression of cardiac MCL-1 in young, diabetic males and females. To test this, we compared cardiac Mcl-1 expression in young rats (ages 11-12 weeks): diabetic males [ZDF obese (n=8)], non-diabetic females [ZDF lean (n=5)], and non-diabetic males [ZDF lean (n=8)] on Purina 5008 diet to young, diabetic female rats [ZDF obese (n=5)] on Research Diets D12468. Body composition analysis by EchoMRI showed that both male and female lean rats had increased muscle mass compared to their obese, diabetic counterparts, whereas obese, diabetic male and female rats had >5-fold increase in body fat than the corresponding lean controls (p<0.05). Fasting plasma insulin levels in lean female rats were lower than that seen in lean male rats; there was a 49-fold increase in diabetic females, and only a 12-fold increase in diabetic males, compared to lean counterparts (p<0.05). Fasting plasma glucose levels increased 2-3 fold in diabetic males and females, compared to lean controls (p<0.05). Quantitative RT-PCR analysis of mRNA isolated from cardiac tissues of lean and diabetic male and female rats showed that there was a 45% decrease in Mcl-1 mRNA in diabetic males compared to lean males, whereas there was an over 90% suppression of Mcl-1 mRNA in diabetic females compared to lean females (p<0.05). Previously, we have shown that insulin improves MCL-1 expression in female mouse cardiomyocytes. Data presented here suggest that although diabetic female rats have more severe hyperinsulinemia compared to diabetic male rats, loss of cardiac MCL-1 expression is more severe in diabetic females compared to diabetic males.This may underlie the loss of cardioprotective benefits in diabetic females.
Abstract 636: Factors Impacting Blood Pressure Control in the Elderly and very Elderly Hypertensive Patients After 8 Weeks Valsartan/Amlodipine SPC Treatment: CHINA STATUS II Subgroup Analysis
Authors: Su Guohai , Li Yueyan , Wang Lei , Wei Min , Cai Xiaojun , Li Zhenhua , Wei Fang , Guan Yuqing , Miao Wei
Published: March 2022

Objective:
To determine the factors that impact BP control in Chinese elderly (aged 65-79 years) and very elderly (aged ≥80 years) hypertensive patients after 8-week treatment with val/aml SPC in the CSII study.

Design and Methods:
3609 elderly and very elderly hypertensive patients uncontrolled by monotherapy with BP ≥140/90 mm Hg (≥130/80 mm Hg for diabetes or CKD) were analyzed. It was a post-hoc analysis of subgroups from the CSII study. All patients received val/aml 80/5 mg SPC for 8 weeks. An additional antihypertensive agent added in patients who were still uncontrolled at the end of 4 weeks. Complications, previous antihypertensive treatment and demographic factors were recorded. Chi-square test, Student’s t-test and logistic regression analysis were conducted.

Results:
After 8 weeks treatment, 61.5% elderly (n=459) and 63.3% very elderly (n=1811) patients achieved BP control. In the 740 elderly patients, low BP control rates were observed in patients with coronary heart disease (CHD) [P <0.05 vs without CHD]. Significant decrease of BP control rates also were observed in 620 elderly type 2 diabetes mellitus (DM) patients (P<0.0001 vs. non-diabetics) and 89 elderly heart failure (HF) patients (P<0.05, vs without HF) while stroke had no effect(P>0.05). Previous antihypertensive treatment (ACEI, ARB, CCB, βblocker and diuretic) had no significant influence on BP control (P>0.05
)
.
Among cardiovascular risk factors
,
while dyslipidemia influenced the BP control rate(P<0.05); patient’s sex, BMI, smoking and alcohol consumption had no effect(P>0.05). In very elderly patients, DM led to low BP control (P<0.0001), but there was no significant impact of CHD, HF and stroke. Previous antihypertensive drugs did not influence the BP control rate (P>0.05), but alcohol consumption had a significant impact (P<0.05).

Conclusion:
CHD, DM, HF and dyslipidemia negatively influenced BP control in the elderly patients. In the very elderly patients, diabetes and history of alcohol consumption were associated with low BP control rates. The effect of the above risk factors on BP control should be considered during the management of hypertension in the elderly and very elderly Chinese patients with above complications as they would need stricter BP control.
Abstract 642: Renal Denervation Prevents Dendritic Cell Activation and Renal T Cell Infiltration and Subsequent Renal Damage in Mice with Angiotensin II-induced Hypertension
Authors: Xiao Liang , Kirabo Annet , Wu Jing , Saleh Mohamed , Zhu Linjue , Harrison David G
Published: March 2022

Hypertension is associated with increased sympathetic outflow and subsequent activation of adaptive immunity. We hypothesized that renal sympathetic nerves activate dendritic cells, and that these in turn cause T cells to proliferate and accumulate and the eventual end-organ damage in the kidney. To test this, we performed bilateral renal denervation (RDN) in C57BL/6 mice by applying phenol to the renal artery and cutting large visible nerves. One week later, mice received an infusion of angiotensin II (490 ng/kg/min) subcutaneous for 14 days. RDN lowered the hypertensive response to angiotensin II infusion (130±3 vs. 161±3 mmHg) as measured by telemetry, and also prevented renal fibrosis, albuminuria and nephrinuria caused by angiotensin as measured by Masson Trichrome stain and ELISA. By flow cytometry, we found that RDN reduced accumulation of total T cells and CD44hi (memory) T cells in the kidney. Angiotensin II infusion increased expression of the maturation marker CD80 in dendritic cells (DCs) of the kidney by 2 to 3-fold, and increased IL-1α, IL-1β, and IL-6 production by splenic DCs by 4 to 6-fold. These increases were attenuated by RDN. We also found that angiotensin II infusion increased VCAM-1 and decreased eNOS and Klf4 mRNA levels in the kidney as measured by real time RT PCR, and these changes were prevented by RDN. We conclude that renal sympathetic nerves contribute to dendritic cell activation, and subsequent T cell infiltration and end-organ damage in the kidney in the development of hypertension. Sympathetic modulation of VCAM-1, eNOS and Klf4 may predispose to inflammatory cell infiltration and renal inflammation.
Abstract 654: Endothelial Ephrin-B2 Essential for Arterial Vasodilation is also Required for Acute Ischemic Recovery
Authors: Lin YuanKai , Jiang Weiya , Youn Ji-Youn , Cai Hua , Wang Rong A
Published: March 2022

Rationale:
The cell surface protein ephrin-B2 is expressed in arterial and not venous endothelial cells throughout development and adulthood. We have recently reported that endothelial ephrin-B2 is essential in acute arterial vasodilation. Here we proposed that endothelial ephrin-B2 plays important protective roles against acute ischemia.

Objective:
We investigated the role of endothelial ephrin-B2 following experimental hindlimb ischemia in mice lacking ephrin-B2 specifically in the endothelium.

Methods and Results:
We performed femoral artery occlusion in mice lacking the ephrin-B2 gene specifically in endothelial cells. We found that the mutant mice exhibited lower residual blood flow than controls immediately following the surgical occlusion by laser doppler perfusion imaging. The mutant mice also developed gangrene not seen in controls soon after the occlusion. Histologically the tissue regeneration and recovery were impaired in the hindlimb of the mutant mice. S-nitrosoglutathione, a donor for nitric oxide, alleviated the perfusion impairment immediately after the occlusion and improved the tissue survival and recovery in the mutants to levels comparable to those in controls. We further demonstrated that acetylcholine-stimulated nitric oxide production was impaired in ephrin-B2 mutant aortae, detected by both chemiluminescence that measures NOx and electron spin resonance that measures bioavailable NO.

Conclusions:
Our data demonstrate that mice lacking ephrin-B2 in endothelial cells are defective in acute ischemic recovery after femoral artery occlusion. Our data also suggest that endothelial ephrin-B2 could potentially interact with the nitric oxide pathway that is critical in its function in arterial dilation and protection against ischemia.
Abstract 656: Efficacy and Safety of Valsartan/Amlodipine Single-Pill Combination in Chinese Elderly Hypertensive Patients with Chronic Kidney Disease: Subgroup analysis of the CHINA STATUS II study
Authors: Wang Wei , Wang Kunlun , Zhou Xiaoyong , Mo Weichun , Song Weiping , Wu Shiqing , Gao Mingqiang , Liu Guoping
Published: March 2022

Objective:
To evaluate the efficacy and safety of Valsartan/Amlodipine SPC in a sub-group analysis of elderly hypertensive patients (aged ≥65 years) with Chronic Kidney Disease (CKD) treated for 8 weeks in the CHINA STATUS II study.

Methods:
This post-hoc analysis was conducted in 3297 hypertensive elderly patients from CHINA STATUS II study(CSII), whose blood pressure (BP) was not controlled by monotherapy . 116 patients had CKD (aged 75.5±6.8 years) while 3181 were without CKD (aged 74.4±6.3 years). All patients in CSII received Valsartan/Amlodipine 80/5 mg SPC for 8 weeks. The patients did not achieve BP control at the end of 4 weeks (BP≥ 130/80 mmHg for patients with diabetes or CKD and≥140/90 mmHg for others), an additional antihypertensive agent added. T -test and chi-square test were used for statistical analysis.

Results:
After valsartan/amlodipine SPC treatment for 8 weeks, mean sitting systolic blood pressure (MSBP) decreased, 25.4±13.3 mmHg and 28.3±12.5 mmHg from baseline in patients with and without CKD, respectively. Mean sitting diastolic blood pressure (MSDP) was reduced by10.4±9.3 mmHg and 13.5±9.5 mmHg from baseline in patients with and without CKD, respectively. 17.2%patients with CKD achieved their BP target of <130/80 mmHg and 77.8% non-CKD patients achieved their BP goal, <140/90 mmHg. The incidence of adverse events was 0.9% in patients without CKD, while none were reported in patients with CKD. Two serious adverse events (SAEs), which were not drug-related, occurred in non-CKD patients and no SAE was reported in CKD patients. 100% of the patients with CKD and 99.2% of patients without CKD rated the tolerance to therapy as ‘good’ or ‘very good’, respectively.

Conclusions:
Valsartan/amlodipine SPC showed good safety and tolerance in hypertensive patients aged ≥65 with CKD whose BP was uncontrolled by previous monotherapy. Lower proportion of elderly patients with CKD achieved the strict BP target compared to elderly patients without CKD. In view of the limitation of this being a post-hoc analysis, the reason for the observed low BP control rate in elderly patients with CKD needs further analysis and research.
Abstract 001: Mutation in the PPARγ Ligand Binding Domain Impairs the Anti-inflammatory Action of PPARγ
Authors: Mukohda Masashi , Ketsawatsomkron Pimonrat , Stump Madeliene , Quelle Frederick W , Sigmund Curt D
Published: March 2022

Peroxisome proliferator-activated receptor gamma (PPARγ) has been proposed to antagonize the activities of nuclear factor kappa B (NFκB) to regulate inflammation. Transgenic mice expressing dominant negative (DN) PPARγ specifically in vascular smooth muscle cells (SMC) exhibited exacerbated atherosclerosis but the mechanism remains unknown. We hypothesized that DN PPARγ promotes NFκB-induced inflammation in SMC. To test this, we cultured mesenteric SMCs from transgenic mice that would conditionally express wild-type (WT) or DN PPARγ (P467L) in response to adenovirus expressing Cre-recombinase (AdCRE). PPARγ expression remained silent in control SMC infected with AdGFP. TNF-α (0.05 ng/ml, 6 hr) induced NFκB target gene (MCP-1, iNOS and MMP9) expression to a greater extent in P467L-Cre compared to P467L-GFP. For example, MMP9 expression was induced 6.3±0.2-fold in P467L-Cre vs 3.2±0.5-fold in P467L-GFP (p<0.01). The NFκB subunit, p65, mRNA level was not altered in these cells. There was no induction of the PPARγ target aP2 in P467L-Cre, but it was induced 6.5±1.7-fold in WT-SMC infected with AdCRE (WT-Cre). The ability of TNF-α to induce NFκB target gene expression was blunted or abrogated in WT-Cre cells, and their expression was significantly reduced in TNF-α-treated WT-Cre compared to WT-GFP (MMP9: 0.7±1.2 vs 6.0±0.3, p<0.01). To examine mechanisms in vivo, we crossbred transgenic mice expressing WT PPARγ specifically in SMC (S-WT) with mice expressing luciferase under control of a NFκB-responsive promoter. TNF-α (500 ng/ml, 24 hr)-induced NFκB activity was decreased in aorta and carotid artery from S-WT mice compared to control mice (aorta: 4.7±1.1 v s 6.7±0.7, p<0.05, carotid artery: 4.0±0.6 vs 8.7±1.2, p<0.01). Finally, to assess the mechanism preventing anti-inflammatory activity by DN PPARγ, we assessed its interaction with p65 protein when co-expressed in HEK293T cells. WT PPARγ co-precipitated with p65, but the interaction between p65 and P467L PPARγ was severely impaired (n=6). Other mutants (R165T, V290M) could bind to p65, suggesting that loss of the ability is specific to P467L PPARγ. We conclude that SMC-PPARγ has anti-inflammatory effects mediated through inhibition of NFκB activity, which is abolished by P467L mutation.
Abstract 006: Distinct and Overlapping Roles of Cytokines IL-17A and IL-17F in Angiotensin II-induced Hypertension and End-organ Injury
Authors: Saleh Mohamed A , Norlander Allison E , Harrison David G , Madhur Meena S
Published: March 2022

Recently, interleukin-17A (IL-17A) has been found to contribute to the renal and vascular dysfunction associated with hypertension, but the mechanisms involved are unknown. We and others found that IL-17F is upregulated in IL-17A deficient mice, suggesting a compensatory response. The goal of the present study was to determine the role of IL-17F in hypertension and the effect of IL-17A or IL-17F neutralization on blood pressure and renal/vascular inflammation. Twelve-week-old male C57BL/6J mice received angiotensin II (Ang II) (490 ng/kg/min, s.c. via 4 wk-osmotic minipump). After 2 weeks, the systolic blood pressure as measured by tail-cuff was 179.4 ± 3.7 mmHg, n=33. Mice were then randomly assigned to one of four treatment groups: (1) Mouse anti-IL-17A; (2) Rat anti-IL-17F; (3) Mouse IgG1 control; (4) Rat IgG1 control. Mice were injected intraperitoneally with 100 μg of antibodies twice weekly during the remaining 2 weeks of Ang II infusion. The administration of anti-IL-17A, but not anti-IL-17F, improved Ang II-induced hypertension (166.5 ± 7.4 mmHg, n=10 vs 181.8 ± 8.6 mmHg, n=7 respectively). Interestingly, both anti-IL-17A and anti-IL-17F blunted the increase in aortic and renal total leukocyte infiltration as quantified by flow cytometry in both organs (Fig. 1a). In addition, both treatments attenuated the increase in glomerular injury as monitored by albuminuria measurement (ELISA) during the Ang II infusion period (Fig. 1b). In conclusion, IL-17F neutralization does not lower blood pressure in response to Ang II, but both IL-17A and IL-17F are pro-inflammatory and orchestrate a major role in the end-organ damage associated with Ang II-induced hypertension.
Abstract 011: Chronic Renal-artery Infusion of Insulin+Glucose Increases Mean Arterial Pressure in Rats
Authors: Irsik Debra L , Duggan A. D , Alaisami Rabei N , Blazer-Yost Bonnie L , Staruschenko Alexander , Brands Michael W
Published: March 2022

Obesity, metabolic syndrome, and Type 2 Diabetes are a continuum of hyperglycemia and hyperinsulinemia that is associated with hypertension. We have reported that inducing chronic hyperglycemia and hyperinsulinemia by continuous IV infusion increases blood pressure in rats. To test the hypothesis that a renal mechanism underlies the increase in blood pressure, we developed a method for chronic renal-artery (IR) infusion in rats. Male Sprague Dawley rats underwent right nephrectomy and a catheter was placed in the left renal artery. Artery and vein catheters and a DSI telemetry unit for 24 hr/day blood pressure also were implanted. Control IV and IR vehicle infusions were begun in all rats via dual-channel Instech swivels. Rats then were assigned randomly to receive either IV (n=11) insulin (1.5 mU/kg/min) and glucose (20 mg/kg/min) or IR (n=8) insulin and glucose at 20% of the IV doses. The alternate syringe in each group continued with the control vehicle solution. MAP averaged 108±1 and 110±2 mmHg in IV and IR rats, respectively, during control, increased progressively and significantly in both groups during 7 days of insulin+glucose infusion, and returned towards control during recovery. Cumulative sodium balance increased significantly in both groups, with no significant change in GFR in either group. Blood glucose did not change significantly and did not differ between groups. Plasma insulin (uU/ml) did not change significantly in the IR rats (8.3±2.6 to 4.8±1.2), but increased significantly in the IV group (13.3±5.5 to 27.9±4.9). These data suggest that chronic hyperinsulinemia and hyperglycemia increase arterial pressure through a direct action on the kidneys.
Abstract 012: Brain Endoplasmic Reticulum (ER) Stress Reduces Appetite and Increases Blood Pressure Independent of the Melanocortin-4 Receptor in Rats
Authors: do Carmo Jussara M , da Silva Alexandre A , Yoo Jung , Moak Sydney P , Spradley Frank , Hall John E
Published: March 2022

Although there is evidence that chronic endoplasmic reticulum (ER) stress affects hypothalamic pathways that regulate food intake, body weight and blood pressure (BP), the specific mechanisms are unclear. One key pathway for controlling energy balance and BP is the central nervous system (CNS) melanocortin system. However, the importance of this system in mediating the effects of ER stress on metabolic and cardiovascular function is unclear. In this study we examined the role of the melanocortin-4 receptor (MC4R) in controlling blood pressure (BP) and metabolic functions during chronic brain ER stress. MC4R knockout (MC4R
-/-
, n=5) and control wild-type Wistar Hannover rats (WT, n=5) were implanted with blood pressure (BP) telemetry transmitters and an intracerebroventricular (ICV) cannula was inserted into the third ventricle at 22 weeks of age. After 10 days of recovery, food intake, BP and HR were measured 24-hrs/day. After stable baseline measurements for 4 days, thapsigargin (TG, 5μg/5μl, ICV) was injected daily for 3 consecutive days to induce ER stress. At baseline, MC4R
-/-
rats ate 23% more food and were 41% heavier than WT rats. MAP was slightly higher (115±3 vs. 109±2 mmHg) and HR was lower (318±10 vs. 363±6 bpm) in MC4R-/- rats. Induction of brain ER stress decreased food intake (26%) while causing no changes in blood glucose levels (WT: 94±4 vs. 98±3 and MC4R-/-: 110±6 vs.116±4 mg/dl) or HR in both groups. Induction of brain ER stress also raised BP in both groups (7 and 5 mmHg, respectively for WT and MC4R
-/-
rats). These results suggest that chronic brain ER stress-induced reductions in appetite and increases in BP are independent of MC4R signaling. (NHLBI-PO1HL51971, NIGMS- P20GM104357 and AHA SDG5680016)
Abstract 018: Sox6 is a Critical Factor in Renin Cell Fate in Development and Pathology
Authors: Gomez Jose
Published: March 2022

Introduction:
the renin-angiotensin system (RAS) is an important component of blood pressure regulation in mammals. Renin, primarily expressed and secreted by kidney Juxtaglomerular (JG) cells, catalyzes the rate limiting step in RAS. However, the transcriptional mechanisms that govern the specification of renin expressing cells in development and under normal or pathophysiological conditions remain poorly understood. We sought to determine new regulators of renin cell fate during kidney development and JG recruitment.

Methods:
Gene expression profiles of renal MSC and JG cells were determined by Affymetrix Mouse 430 2.0 array. JG cells and renal MSC were isolated from adult C57Bl6 Ren1c YFP mice by collagenase digestion of the kidney, followed by flow cytometry to select for the cells. Renin expression
in vitro
was induced by treatment with IBMX and Forskolin. Renal MSCs were transduced with lentivirus carrying vectors for Sox6, Sox6 shRNA or controls.
Ex vivo
analysis was performed in embryonic kidneys (14.5 dpc) isolated and transduced with Sox6 shRNA or scrambled shRNA. The kidneys were then cultured for 4 days.

Results:
Microarray data showed that the transcription factor Sox6 was highly expressed in FACS isolated JG cells compared to renal MSC (96-fold, n=3, P<0.05). This finding was validated by qPCR (100-fold, n=4, P<0.05). Sox6 was expressed in renin producing cells during kidney embryogenesis as determined by immunofluorescence and confocal microscopy (E14.5, N=4). Knockdown of Sox6 by shRNA in an
ex vivo
model of kidney development resulted in a 70% reduction of renin expression (N=4, P<0.01). Differentiation of adult renal MSCs to renin producing cells in vitro was enhanced by overexpression of Sox6 (20-fold, N=6, P<0.01). Knockdown of Sox6 by shRNA inhibited in vitro MSC differentiation (5-fold, N=6, P<0.001). In vivo, low salt and furosemide, which stimulates JG recruitment, increased Sox6 expression (5-fold, N=5, P<0.001) and colocalization with renin.

Conclusion:
Our results support a novel and critical role for the transcription factor Sox6 in renin cell fate and in renal development and physiology. Further studies can provide an in-depth understanding of the role of Sox 6 in hypertension and its therapy.
Abstract 029: Reduced Bone Marrow Adrenergic Receptor Signaling Modulates Inflammatory Factors and Alters Gut Microbiota
Authors: Yang Tao , Li Eric , Ahmari Niousha , Carvajal Jessica , Santisteban Monica , Kim Seungbum , Raizada Mohan , Zubcevic Jasenka
Published: March 2022

Brain - bone marrow (BM) communication is implicated in regulation of blood pressure (BP) in neurogenic hypertension (HTN), and chronic elevation of sympathetic drive contributes to BM inflammatory cell (IC) activity and promotes vascular inflammation. Our recent study linked gut dysbiosis to human and animal HTN; thus, we hypothesize that blocking the effects of sympathetic drive in the BM will result in reduction of BM ICs and modify gut microbiota.
Methods
: Whole BM cells were extracted from adrenergic receptor beta 1 and 2 knock out mice (Adrb1
tm1Bkk
Adrb2
tm1Bkk
/J KO) and reconstituted into lethally- irradiated C57BL/6J mice to generate C57-AdrB1.B2 KO chimera, characterized by reduced/diminished effect of sympathetic drive on the BM cells. Control mice (C57-C57) were generated by reconstitution of irradiated C57BL/6J with whole BM from C57BL/6J mice. All mice were recovered for 2-3 months prior to measurements. Specific primers were used to amplify variable region 4-5 of 16S rDNA from isolated fecal DNA. Purified amplicons were pooled to generate bacterial library, quantified by qPCR and subjected to Illumina Miseq sequencing. Reads were aligned with the Silva nonredundant 16S reference database. BP recordings were performed using radiotelemetry (DSI). Blood was collected at Night (8-9pm, i.e. at highest activity) for analysis of circulating ICs.
Results
: We observed a significant reduction of circulating ICs (CD4
+
CD8
+
: 2.3±0.6% vs. 0.876±0.1%; CD11b
+
: 15.4±4% vs. 8.5±0.8%; CD184
+
CD62L
+
: 25±4.3% vs. 6.6±0.8%; (% of mononuclear cells)), and increase in endothelial progenitors (Tie2
+
: 21.3±1.5% vs. 32.2±5.7%) in chimera vs. control. A trend in BP reduction was observed in chimera (Day: 110±6 vs. 101±2 mm Hg; Night: 130±2 vs. 123±2 mm Hg). Expansion of
Bacteroidetes
(control: 34.2%±3.0 vs. chimera: 49.9%±3.3), and contraction of
Firmicutes
(control: 63.9%±3.0 vs. chimera: 48.9%±3.8) were observed in chimera, lowering the F/B ratio two fold.
Conclusion
: Diminished effect of BM sympathetic drive dampens BM-derived ICs and favors the bloom of Bacteroidetes phylum, suggesting the role of BM in gut homeostasis.
Abstract 048: Epigenetic Regulation of the Human Angiotensinogen Gene: Role of Individual SNPs and Implication for Hypertension
Authors: Mopidevi Brahmaraju , Perla Sravankumar , Puri Nitin , Kumar Ashok
Published: March 2022

Hypertension is a complex disease caused by a combination of genetic and environmental factors. Renin angiotensin system (RAS) dysfunction is a frequent accompaniment of essential hypertension. In this regard, polymorphisms in the angiotensinogen (AGT) gene increase RAS activity and cause blood pressure elevation. We have identified two distinct haplotypes of the human AGT gene constituted by a group of four SNPs in linkage disequilibrium. Variants -1670A, -1562C, and -1561T always occur with -6A and form the haplotype-I (Hap-I), while variants -1670G, -1562G, and -1561G always occur with -6G and constitute haplotype-II (Hap-II). We hypothesized that these SNPs, when present together as Hap-I or Hap-II in transgenic mice (TG) may lead to haplotype-dependent DNA methylation of CpG sites in the promoter of the hAGT gene. Methylation patterns alter gene transcription at an epigenetic level and this, in turn, could be dependent on individual polymorphisms. hAGT promoter CpG sites in the kidney are more methylated as compared to liver and fat. In the Kidney, the hAGT promoter CpG sites are methylated at -460, -434, -386, -346, -282, -261, -245, -229, -218, -185, -144, -18, -11, -7, +10, +42, +65 in Hap-II whereas, in Hap-I the promoter DNA methylation is observed at -460, -434, -401, -386 positions. Further, in both liver and fat the CpG sites were methylated at -11, +42, +65 positions in Hap-II TG mice, whereas in the Hap-I there were no methylation sites detected. The hAGT gene mRNA levels from these tissues were quantitated by qPCR and were 2.1 (kidney), 2.5 (liver) and 4.28 (fat) fold higher in haplotype-I vs. haplotype-II (p<0.05). Our results indicate that, in Hap-II TG mice liver, kidney and fat tissues having more CpG sites methylated in the promoter of the hAGT, and thus having less gene expression. On the other hand, Hap-I has either less or no methylation of CpG sites in the hAGT promoter and hence high gene expression. Thus, we show here for the first time that SNP blocks in the hAGT gene alter its methylation pattern in a tissue-dependent manner. This shifts the paradigm in favor of an interdependent system where epigenetic regulation is reliant on the gene haplotypes; together, these regulate protein expression and associated physiological outcomes.
Abstract 062: Inhibition of Phosphodiesterase 5 Attenuates Angiotensin II Dependent Hypertension and Renal Vascular Dysfunction in C57BL/6 but Not in eNOS-KO Mice
Authors: Thieme Manuel , Sivritas Sema , Potthoff Sebastian A , Yang Guang , Rump Lars C , Stegbauer Johannes
Published: March 2022

Angiotensin (Ang) II is a key player in the pathogenesis of hypertension by influencing the NO/cGMP signaling cascade. Thus, Ang II induces vascular dysfunction by activating phosphodiesterase (PDE) 1 and PDE5 mediated cGMP-degradation. PDE1 is activated by increased intracellular Ca2+ concentrations observed during Ang II mediated vasoconstriction. PDE5 is activated by a cGMP mediated negative feedback mechanism. The present study examines the role of PDE1 and PDE5 in regulation of renal hemodynamics and blood pressure (BP) during Ang II induced hypertension.

In unconscious C57BL/6 (WT) mice, changes in renal blood flow (RBF) and BP was measured in the presence of specific PDE5- (sildenafil) or PDE1- (vinpocetine) inhibitors. Secondly, in WT and eNOS-KO mice, a mouse model characterized by low NO/cGMP levels, hypertension was induced by chronic Ang II infusion (500 ng/kg/min) for 14 days. Vascular relaxation was tested in the presence of acute or chronic PDE1 or PDE5 inhibition in isolated perfused kidneys.

During acute Ang II infusion, sildenafil increased RBF and decreased BP more potently than under baseline conditions. In contrast, vinpocetin had no effect on RBF and BP during normotensive or hypertensive conditions, Moreover, sildenafil but not vinpocetine improved NO (GSNO) dependent vasodilation in isolated perfused kidneys of Ang II treated WT mice. Next, we tested the impact of chronic PDE5 inhibition on Ang II dependent hypertension. Sildenafil (100mg/kg/d) attenuated Ang II dependent hypertension in WT (156±4 vs. 139±7mmHg; p<0.05) but not in eNOS-KO mice (166±4 vs. 172±4mmHg). Respectively, chronic Sildenafil treatment improved NO dependent vasodilation in isolated perfused kidneys of Ang II treated WT but not in eNOS-KO mice. In addition, urinary nitrite excretion, a marker for NO generation was significantly increased during Ang II-infusion compared to baseline in WT but not in eNOS-KO mice.

In conclusion, PDE5 seems to be the predominant PDE regulating RBF and renal vascular function. Moreover, sildenafil ameliorates Ang II dependent hypertension and improves vascular dysfunction. No effect of PDE5 inhibiton was observed in eNOS-KO mice supporting the hypothesis that Ang II activates PDE5 initially by increased NO/cGMP generation.
Abstract 080: Role of High-mobility Group Box 1 in Angiotensin Converting Enzyme 2 -Mediated Cardioprotective Effects in a Mouse Model of Myocardial Ischemia
Authors: Qi Yanfei , Zhang Juan , Wang Lei , Krause Eric , Oh S. P , Shenoy Vinayak , Guzzo Dominic S , Katovich Michael J , Raizada Mohan K , Pepine Carl J
Published: March 2022

Backgrounds:
High-mobility group box 1 (HMGB1) activates inflammation process, and its elevation is associated with adverse clinical outcomes in patients with myocardial infarction (MI). Previous studies have demonstrated that angiotensin-converting enzyme 2 (ACE2), a key member of vasoprotective axis of the renin angiotensin system, provides protection against MI. However, the involvement of HMGB1 in ACE2-mediated protective effects in cardiovascular diseases has yet to be elucidated. Thus, we hypothesized that the cardioprotective effects of ACE2 are, in part, by inhibition of cardiac HMGB1 and inflammatory pathways.

Methods and results:
Transgenic (TG) mice that ubiquitously overexpress ACE2 were used for the study. ACE2 levels in the hearts of TG mice were 58-times higher than wild type (WT). Mice were subjected to coronary artery ligation surgery and cardiac functions were evaluated by echocardiography 4 weeks post MI. MI resulted in a 53% decrease in ejection fraction (EF) and a 42% infarct area of left free wall. However, ACE2 TG mice showed 60% improvement of EF over WT mice. This was associated with a 55% decrease in the infarct area. Additionally, MI caused a three times increase in plasma HMGB1 levels which was associated with elevated macrophage infiltration and proinflammatory cytokine levels. In comparison to WT-MI mice, ACE2 overexpression attenuated the increase in plasma HMGB1 by 25% and reduced the proinflammatory cytokine levels to normal in the TG mice. To investigate the modulatory effects of ACE2 on HMGB1, adult cardiomyocytes incubated with an ACE2 activator (DIZE), a selective ACE2 inhibitor (C16), and a combination of DIZE and C16 were exposed to hypoxic condition for three hours. Hypoxia (H) caused 35% cell death and 44% increase in the HMGB1 level (H: 23.6± 0.8 vs normoxia: 16.4± 1.2, ng/ml). DIZE significantly attenuated the cell death (2% cell death) and 37% decrease in HMGB1 levels (DIZE+H: 14.8± 2.5 ng/ml), which was blocked by a selective ACE2 inhibitor (C16+DIZE+H: 20.8± 2.9ng/ml).

Conclusions:
The protective effect of ACE2 against ischemia is correlated with decrease in HMGB1 and its downstream pro-inflammatory cascades. Thus, HMGB1 could be useful target for the development of novel treatment for MI.
Abstract 118: Role of Connecting Tubule Glomerular Feedback in Tubuloglomerular Feedback Resetting After Unilateral Nephrectomy
Authors: Monu Sumit R , Wang Hong , Ren Yilin , Kutskill Kristopher , Romero Cesar , Juncos Juan Xavier , Janic Branislava , Peterson Ed , Garvin Jeffrey L , Carretero Oscar A
Published: March 2022

Tubuloglomerular feedback (TGF) and connecting tubule glomerular feedback (CTGF) autoregulate nephronal afferent arteriolar resistance. In TGF, the macula densa signals the afferent arteriole to constrict when NaCl transport is enhanced by increased luminal NaCl, via sodium[[Unable to Display Character: –]]potassium-2-chloride cotransporter-2 (NKCC2). CTGF is mediated by connecting tubule sodium transport via epithelial sodium channel (ENaC) and dilates the afferent arteriole. Attenuation or resetting of TGF occurs after unilateral nephrectomy (UNX), but the mechanism behind this resetting remains unclear. This TGF resetting after UNX has been implicated in progressive glomerular damage due to sustained increase in glomerular capillary pressure. Since TGF is attenuated after UNX, we sought to test the hypothesis that CTGF is enhanced and that it contributes to TGF resetting after UNX. To test this hypothesis, we performed right side UNX in Sprague Dawley (SD) rats. 24 hours after surgery, we performed micropuncture of individual rat nephrons while measuring stop-flow pressure (PSF), which is an index of glomerular capillary pressure and afferent arteriolar tone. PSF decreases with an increase in afferent arteriolar tone. TGF response was measured as a decrease in PSF induced by switching late proximal perfusion from 0,10,20,30 and 40nl/min. Maximal TGF response was 1.3 ± 1.7 mmHg in UNX rats while 8.2 ± 0.9 mmHg in sham-UNX rats indicating a TGF resetting in UNX rats. When CTGF was inhibited with the ENaC blocker Benzamil (1μM), TGF response was 10±1.2 mmHg in UNX rats and 14.8± 1.3 mmHg in sham-UNX rats, indicating the restoration of TGF responses in UNX. We conclude that enhanced CTGF contributes to the TGF resetting after 24 hours of UNX. Enhanced CTGF may be responsible for glomerular damage post UNX.
Abstract 119: The Imbalance of Mitochondrial Superoxide Dismutase Activity and Superoxide Production in Endothelial Dysfunction and Hypertension
Authors: Dikalova Anna E , McMaster William G , Itani Hana A , Nazarewicz Rafal R , Fessel Joshua P , Gamboa Jorge L , Harrison David G , Dikalov Sergey
Published: March 2022

Clinical studies have shown that Sirt3 expression declines by 40% by age 65 paralleling the increased incidence of hypertension and metabolic conditions further inactivate Sirt3 due to increased NADH and Acetyl-CoA. Sirt3 activates a major mitochondrial antioxidant enzyme, superoxide dismutase 2 (SOD2) by deacetylation of specific lysine residues. We hypothesized that loss of Sirt3 activity increases vascular oxidative stress due to SOD2 hyperacetylation and this promotes endothelial dysfunction and hypertension. Indeed, Western blot analysis showed 3-fold increase in SOD2 acetylation and 1.5-fold decrease in Sirt3 level in hypertensive human subjects. Infusion of angiotensin II (0.7 mg/kg/day) in C57Bl/6J mice increased acetylation of vascular SOD2 by 2-fold, reduced SOD2 activity to 52% and raised blood pressure to 156 mm Hg. We have tested if Sirt3 depletion exacerbates endothelial dysfunction and hypertension. Indeed, Western blot analysis of Sirt3-/- mice showed SOD2 hyperacetylation and reduced SOD2 activity, however, blood pressure and superoxide production was normal. Angiotensin II infusion in Sirt3-/- mice increased blood pressure to 182 mm Hg, increased superoxide, reduced endothelial nitric oxide and impaired acetylcholine-mediated vasodilatation compared with angiotensin II infused wild type mice. Treatment of hypertensive Sirt3-/- mice with SOD2 mimetic mitoTEMPO (1.4 mg/kg/day) after onset of angiotensin II-induced hypertension normalized blood pressure and restored endothelial dependent vasodilatation. We suggest that mitochondrial superoxide does not contribute to the basal vasodilation and blood pressure regulation however high salt, angiotensin II and inflammation increase mitochondrial superoxide. Indeed, superoxide production by complex I in submitochondrial particles was significantly increased in hypertensive mice and this was associated with complex I S-glutathionylation. Interestingly, SOD2 overexpression in transgenic mice prevents complex I S-glutathionylation, reduces superoxide production by complex I and attenuates hypertension. These data indicate that imbalance of SOD2 activity and mitochondrial superoxide production contributes to endothelial dysfunction and hypertension.
Abstract MP01: Vascular versus Tubular Renin: Role in Kidney Development
Authors: Sequeira Lopez Maria Luisa S , Nagalakshmi Vidya K , Li Minghong , Sigmund Curt D , Gomez R. Ariel
Published: March 2022

Renin, the key regulated enzyme of the renin angiotensin system regulates blood pressure and fluid electrolyte homeostasis as well as morphogenesis of the kidney. Classically, renin is synthesized and released from juxtaglomerular cells located in the afferent arterioles at the entrance to the glomeruli. It has also been suggested that renin may also be synthesized by tubular cells. Interestingly, whole body deletion of the renin gene results in striking vascular and tubular abnormalities, hydronephrosis and a urine-concentrating defect. Given the complexity of such phenotype, it has not been possible to discriminate the relative contributions of vascular versus tubular renin. To investigate the relative contribution of renin in the vascular versus tubular compartments during kidney development we deleted renin independently in either compartment by crossing Ren1cfl/fl mice to Foxd1-cre or to Hoxb7-cre mice. We performed blood chemistries, histological analysis, immunostaining for specific cell markers, total kidney renin mRNA quantification and ELISA for renin in plasma. Vascular deletion of renin (Foxd1 lineage) resulted in neonatal mortality that could be rescued with daily injections of saline suggesting that this phenotype is due to an inability to concentrate the urine and improper medullary development. These mice showed absence of renal renin and negligible levels of plasma renin. Histologically, the kidneys had abnormal development of its arterioles, which became progressively thickened by disorganized, concentric hypertrophy of smooth muscle cells and marked hydronephrosis. On the other hand, lack of renin in the collecting ducts (Hoxb7 descendants) did not affect kidney morphology, intra-renal renin distribution, kidney renin mRNA expression or circulating renin during basal conditions or in response to a homeostatic stress such as low sodium diet. We conclude that renin generated in the renal vascular compartment is fundamental for the development and integrity of the kidney whereas the presence of renin in the collecting duct system is dispensable for normal kidney development and cannot compensate for the lack of renin in the vascular compartment. Further, the main source of circulating renin is the kidney vasculature.
Abstract P002: Aldosterone Promotes the Release of miRNA-Containing Exosomes from Endothelial Cells, Leading to Uptake by Smooth Muscle Cells
Authors: Robertson Stacy , Romano Adelina , Dababneh Edward , Bursill Christina
Published: March 2022

Background:
Aldosterone increases exocytosis from endothelial cells, which may contribute to the detrimental actions of aldosterone in the vascular system. We propose a role for exocytosis of exosomes; these small extracellular particles contain host cell genetic material and once released they can circulate in the blood and transfer their contents (e.g. miRNAs) to recipient cells. We aimed to examine the effect of aldosterone on exosome release from endothelial cells, the miRNA content of exosomes and finally, exosome trafficking of miRNA from endothelial cells to smooth muscle cells.

Methods and Results:
Exosomes were isolated via ultracentrifugation from the media of human coronary artery endothelial cells (HCAECs) following incubation with aldosterone (100nM) or vehicle (DMSO). Exosomes were analyzed using western blot analysis testing for exosome and cell markers, and by nanoparticle-tracking analysis (NanoSight); aldosterone increased exosome concentration by 2.65 fold (19.1x1010 particles/mL) compared to the vehicle control (7.2x1010 particles/mL). RNA was extracted from isolated exosomes and host HCAECs and levels of miR-24 measured by qRT-PCR. miR-24 levels were unaffected by aldosterone stimulation in HCAECs (0.58±0.19; p=0.07) but were significantly increased in exosomes isolated from the media (2.35±0.07 fold; p=0.03). To test exosome paracrine trafficking, HCAECs were transfected with a c.elegans miRNA (cel-miR-39) and incubated adjacent to human coronary artery smooth muscle cells (HCASMCs) separated by a 0.4 μm filter, with or without aldosterone. Levels of cel-miR-39 in HCASMCs increased significantly (2.42±0.52 fold; p=0.02) in aldosterone-stimulated cells, compared to control. Blocking transcription with Dactinomycin (2.07±0.61 fold; p>0.05) or blocking the mineralocorticoid receptor had no effect on aldosterone-mediated trafficking (3.43±1.49 fold; p>0.05).

Conclusion:
This study demonstrates that aldosterone: 1) increases exosome secretion from endothelial cells, 2) increases miR-24 content of exosomes and 3) contributes to an increased uptake of exosome-packaged microRNAs by smooth muscle cells. Together, this represents a novel mechanism by which aldosterone contributes to vascular disease.
Abstract P023: Toll-like Receptor 4 Deficiency Reduces Macrophage Mediated Renal Injury in Ang-II Induced Hypertension
Authors: Pushpakumar Sathnur , Amin Matthew , Sen Utpal
Published: March 2022

Infiltration of macrophages has been amply demonstrated in the kidney of humans and experimental models of hypertension however, its role in tissue injury and repair remains unclear. Furthermore, recent studies implicate upregulation of toll-like receptor 4 (TLR4) and inflammation in hypertension and vascular remodeling. Previously, we observed that C3H/HeJ mice lacking functional TLR4 were hyporesponsive to Ang-II induced hypertension and renal damage. In this study, we hypothesized that TLR4 deficiency inhibits macrophage mediated inflammation and TGF-β-induced fibroblast accumulation to reduce renovascular injury and fibrosis. Male C3H/HeOuJ (functional TLR4) and C3H/HeJ mice (dysfunctional TLR4) aged 8-10 weeks underwent subcutaneous osmotic pump insertion for infusion of Angiotensin-II (Ang-II) at 1000 ng/kg/min or vehicle (0.9% saline) for 4 weeks. Blood pressure (BP) was measured twice weekly by non-invasive tail-cuff plethysmography. Ang-II treatment increased mean BP in both groups but to a greater degree in C3H/HeOuJ mice compared to C3H/HeJ (138.6± 6.3 vs. 103.4±7.2 mm/Hg). Plasma creatinine in saline treated C3H/HeOuJ and C3H/HeJ mice was normal (0.37±0.026 and 0.32±0.023 mg/dL respectively) in contrast to ang-II treated mice (1.34±0.075 vs. 0.78±0.052 mg/dL). Flow cytometry revealed increased F4/80+ and CD40+ cells indicating pro-inflammatory M1 macrophage in Ang-II treated C3H/HeOuJ compared to C3H/HeJ mice. Similarly, Ang-II increased the mRNA and protein expression of CD40, MCP-1, TNF-α and TGF-β in C3H/HeOuJ compared to C3H/HeJ mice. Immunostaining revealed increased colocalization of CD45 and procollagen-1 in renal sections of C3H/HeOuJ mice suggesting increased accumulation of myeloid fibroblasts than in C3H/HeJ mice. The expression of α-smooth muscle actin an indicator of myofibroblast and extracellular matrix proteins, collagen type I and fibronectin (Western blot and immunostaining) was increased in C3H/HeOuJ compared to C3H/HeJ mice. In conclusion, our data suggests that in C3H/HeJ mice, deficiency of functional TLR4 reduces macrophage activation and inflammation which in turn decreases TGF-β-induced renal fibrotic response in Ang-II induced hypertension.