bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P007: Hypertension and Myocardial Mechanics in Patients with Hypertrophic Cardiomyopathy
Authors: Manocha Kevin K , Snipelisky David F
Published: March 2022

Introduction:
Strain abnormalities in patients with Hypertrophic Cardiomyopathy (HCM) is associated with poor prognosis. Our aim was to assess whether the presence of hypertension in HCM altered myocardial mechanics using strain echocardiographic imaging.

Hypothesis:
Hypertension is a risk factor for diminished deformation in patients with HCM.

Methods:
A retrospective chart review of patients undergoing HCM evaluation at our facility between December 2008 and December 2011 was performed. Patients were divided into subsets based on the presence of hypertension as a documented diagnosis. All patients had comprehensive echocardiographic assessments with strain imaging. Clinical data was extracted from the electronic medical record. Comparisons in strain values were made between both subsets.

Results:
A total of 200 patients was assessed. The average age was 56 years (60% male) and 42.5% of the patients had a diagnosis of hypertension. Global longitudinal strain, Mid strain, Basal strain, Anterior, Lateral, Inferior, Inferolateral strain had no relation to the presence of hypertension. Septal strain (mean without hypertension -14.306 vs with hypertension -16.071, p value: .0069) Inferoseptal strain (mean without hypertension -12.813 vs with hypertension -15, p value: .0018), and Anteroseptal deformation (mean without hypertension -15.301 vs with hypertension -16.784, p value: .0445) however, was increased in patients with hypertension compared to patients with no history of hypertension (See Table)

Conclusions:
The presence of hypertension is associated with septal strain abnormalities. Thus, future exploration of hypertension control and hypertrophic cardiomyopathy prognosis and outcomes need to be explored.
Abstract P009: Relationship Between Increased Systolic and Diastolic Pressure and Left Atrial Train Rate
Authors: Yang Tsung Yeh , Hung Ta-Chuan , Kuo Jen-Yuan , Hou Jia-Yin , Yeh Hung-I , Hung Chung-Lieh
Published: March 2022

Background:
Left atrial (LA) strain rate analysis by two-dimensional speckle tracking can represent a new tool to evaluate LA function. Office blood pressure is the most powerful non-invasive index of arterial stiffness which can determine LA function.

We hypothesized that increased systolic and diastolic pressure were associated with increased left atrial strain.

Methods:
We consecutively examined 2D speckle-tracking in an asymptomatic cohort, with cardiac geometry, LA volume, LA ejection fraction, LA strain rate analyzed. The LA speckle tracking echocardiography curves were obtained using R-wave onset of the electrocardiogram as a reference point. The LA strain rate (SRs, SRe and SRa) were analyzed by commercialized software (GE EchoPAC; GE Vingmed, Norway).

Results:
A total of 4052 volunteers were divided into quintiles by systolic or diastolic blood pressure (systolic pressure cut point 110, 118, 126, 136mmHg; diastolic pressure cur=t point 68, 71, 80, 85mmHg). A trend toward the greater systolic or diastolic pressure, the higher LA diameter, LA volume and LA strain rate across 5 groups were observed (Table1 and Table 2, all trend p<0.001 except LA SRs).

Conclusion:
Increased in systolic and diastolic pressure were independently associated with increased LA strain rate, LA diameter and LA volume in asymptomatic populations.
Abstract P011: Source and Catabolism Sites of the Cardio-renal Protective Peptide N-Acetyl-Seryl-Aspartyl-Lysyl-Proline (Ac-SDKP)
Authors: Romero Cesar A , Kumar Nitin , Rhaleb Nour-Eddine , Carretero Oscar A
Published: March 2022

Ac-SDKP is a natural tetrapeptide with anti-fibrotic and anti-inflammatory properties in vascular, myocardial and kidney tissues. It is degraded by angiotensin converting enzyme (ACE) and treatment with ACE inhibitors increases its plasma levels. However the main sites of Ac-SDKP production and catabolism are unknown. Thus, the aim of this study was to determine the source and catabolism sites of Ac-SDKP. Ac-SDKP was measured in brain, thymus, lungs, heart, spleen, kidney, liver, intestine, and bone marrow from male 200-250 gr Sprague Dawley rats. Samples from arterial blood delivered to and venous blood exiting from kidney, spleen, intestine, liver, and lungs were obtained, and the vein to arterial (V-A) plasma levels differences were calculated as an index of organ production or elimination. Additionally, urinary Ac-SDKP excretion over 24 hrs and Ac-SDKP renal clearance were measured. Inulin clearance was also determined. The Stop Flow technique was performed to evaluate the renal handling of Ac-SDKP in different segments of the nephron. The greatest amounts of Ac-SDKP were found in Thymus, Spleen and Bone marrow (111.3±9; 92.2±24; 70.3±17 pmoles/mg of protein respectively), while the lowest values were found in brain, large intestine and heart (10.7±6; 14.8±6 ;19.8±7; pmoles/mg of protein, respectively). Ac-SDKP is secreted by different organs into the circulating blood. The highest V-A difference was found in the spleen (15.2±7 nM), reflecting its production, while the lowest V-A differences was observed in the lungs (-1.2±3.7 nM), reflecting a high degradation of Ac-SDKP in this tissue. No V-A differences were found in kidney (0.98±7 nM). It was not possible to obtain venous samples from Thymus or Bone marrow. The renal clearance of Ac-SDKP was 0.15 ml/min/100g body weight, equivalent to a fractional excretion of 14.5%. The stop flow technique showed the highest Ac-SDKP levels in the distal part of the nephron, while the proximal segments showed the lowest (11 Vs. 4.2 Ac-SDKP/Inulin ratio). In conclusion, Ac-SDKP production occurs mainly in lymphoid organs. Expectedly, high Ac-SDKP degradation was observed in the lungs. In the kidney Ac-SDKP is filtered, probably degraded in the proximal tubule and secreted in the distal tubule.
Abstract P012: Mycophenolate Mofetil Prevents Cerebrovascular Injury in Stroke-prone Spontaneously Hypertensive Rats
Authors: Dhande Isha S , Zhu Yaming , Doris Peter A
Published: March 2022

The stroke-prone spontaneously hypertensive rat strain (SHR-A3) develops hypertensive end-organ injury, including strokes and progressive kidney disease, as a result of naturally occurring genetic variations. Our recent work identifies genetic variants in immune signaling pathways that contribute to susceptibility to end organ injury. These variants are present in SHR-A3 animals, but absent in SHR lines that resist end organ injury. To test the hypothesis that a dysregulated immune response promotes stroke susceptibility, 20-week old male SHR-A3 rats were salt loaded (1% NaCl in drinking water) and treated daily with the immunosuppressant mycophenolate mofetil (MMF, 25 mg/kg/day, p.o.) (n=8) or vehicle (saline) (n=9) for 8 weeks. Blood pressure was measured weekly by telemetry for the duration of the study. Compared to untreated controls, MMF-treated SHR-A3 had improved survival and significantly lower neurological deficit scores (1.44 vs 0.125; p<0.02). Salt loading resulted in a progressive increase in blood pressure, which was prevented in rats receiving MMF. At the time of euthanasia, mean systolic BP in the untreated group was 243.0 ± 5.6 mmHg compared to 213.6 ± 7.1 mmHg (p<0.005) in the MMF-treated SHR-A3 group. Gross morphology of the brain revealed cerebrovascular lesions including cerebral edema in 90% (8 out of 9) and microbleeds and hemorrhages in about 50% (5 out of 9) of the untreated SHR-A3 rats. These lesions were absent in the MMF-treated cohort. The expression of CD68, a marker of activated microglia/macrophages, was up-regulated in the brain sections from vehicle-treated rats with microbleeds and hemorrhages, but was not detectable in the brains of rats receiving MMF. MMF also prevented renal injury in SHR-A3 rats, evidenced by reduced proteinuria (albumin to creatinine ratio) from 7.52 to 1.05 mg/mg (p<0.03) and lower tubulointerstitial injury scores (2.46 vs 1.43; p<0.01). Our findings provide evidence that suppression of immune activation prevents the occurrence of cerebrovascular and renal injury in salt-loaded SHR-A3 rats.
Abstract P026: Relationship Between Antihypertensive Adherence and Emergency Department Blood Pressure
Authors: McNaughton Candace D , Rothman Russell L , Brown Nancy J , Roumie Christianne L
Published: March 2022

Background:
Blood pressure is often uncontrolled in the Emergency Department (ED), but the contribution of medication non-adherence to this is not known.

Methods:
Three hundred ED patients with hypertension were enrolled from June 2012 to April 2013 to evaluate the association of antihypertensive adherence (measured by serum assay and by survey) with systolic blood pressure (SBP) in the ED. Presence of anti-hypertensive medications in the blood was detected using a previously validated mass spectrometric assay. Adherence was defined as the ratio of detected to prescribed antihypertensive medications (1=completely adherent, 0=completely non-adherent, <1=partially non-adherent). Patients also completed the Adherence to Refills and Medications Scale (ARMS, where a score of 12 defines adherence and >12 indicates some degree of non-adherence). Spearman’s rho was used to evaluate correlation of the assay and ARMS. A single BP was measured by research staff using an oscillometric cuff with the patient in a seated position. Multiple linear regression was used to assess the relationship between adherence and SBP, adjusting for age, sex, race, insurance, literacy/numeracy, education, and comorbid conditions. Missing numeracy for 18 subjects was imputed.

Results:
Three participants failed to meet inclusion criteria. Remaining 297 patients were 59.1±11.2 (SD) years old, 53.9% female, 62.3% White, 37.7% diabetic, and 24.6% had chronic kidney disease. Patients were prescribed a median of 2 medications (interquartile range 1 to 3, maximum 5). Mean SBP was 136.7±24.2 mmHg. By the assay, 214 (72.1%) patients were adherent, 59 (19.9%) partially non-adherent, and 24 (8.0%) completely non-adherent. By ARMS, 65 (21.9%) were adherent, 232 (78.1%) were non-adherent (rho=-0.23, p<0.001). Adherence by assay was associated with 11.6mmHg lower SBP (95% CI -17.7 to -5.4 mmHg, p<0.001) compared to partial or complete non-adherence. Adherence (by ARMS) was associated with 5.9 mmHg lower SBP, but this was not statistically significant (95% CI -12.6 to 0.8 mmHg, p=0.08).

Conclusions:
In this preliminary study, antihypertensive adherence measured by a mass spectrometry assay was associated with lower SBP among patients in the ED. The assay may be a useful tool for future research.
Abstract P030: Early Cognitive Deficits Induced by Diabetes Are Masked by an Increase in Exploratory Behavior: Evidence From TLR2 Knock-out Mice
Authors: Hardigan Trevor , Hernandez Caterina , Ergul Adviye
Published: March 2022

Toll-like receptor 2 (TLR2) has been shown to contribute to cardiovascular complications of diabetes such as nephropathy. However, the role of TLR2 in cerebrovascular inflammation and dysfunction leading to cognitive impairment, an emerging cerebrovascular complication of diabetes and hypertension, remains unknown. We reported that streptozotocin (STZ)-induced type-1 diabetic TLR2 knockout mice were protected from decreased cerebral blood flow (CBF) at 4 and 6 weeks post induction. Given that decreased CBF precedes cognitive impairment, we hypothesized that these TLR2-KO STZ mice would subsequently be protected from cognitive impairment. 10 week old male C57Bl:6 and TLR2-KO mice were injected with 50mg STZ/kg body weight daily for 5 days and observed alongside control mice. Fasting blood glucose was measured, with levels > 240 mg/dL considered to confirm diabetes. Cognitive function was assessed via Y-maze testing. There was no difference in total arm entries between WT and WT-STZ mice or between WT and TLR2-KO. There was a significant decrease in total entries between TLR2-KO and TLR2-KO STZ (40.33± 1.88 vs.29.67± 2.09, p≥0.05, n=12/group). However, percent novel arm entries was statistically non-significant amongst all groups. Paradoxically, both diabetic groups had a non-significant increase in percent of time spent in the novel arm vs. their respective controls (WT-STZ: 35.7±5.3 seconds vs. WT: 33.2± 4.4 seconds; TLR2-KO STZ: 41.6±5.8 seconds vs. TLR2-KO: 37.9±5.5 seconds). The lack of difference in percent novel arm entries is likely a result of the relative increase in exploratory behavior in the STZ groups. Additional experiments with open-field testing and novel object recognition test are being performed to determine if the apparent lack of change in short term hippocampal memory is due to a confounding artifact of the exploration time and whether these cognitive tests are accompanied by changes in CBF in this cohort. If indeed TLR2 KO mice are protected from early mild cognitive deficits, TLR2 antagonism may be a novel target in the prevention/treatment of cerebrovascular complications of diabetes.
Abstract P041: Inhibition of MicroRNA-221 by Estradiol Contributes to its Differential Effects on Smooth Muscle Cell Growth and Endothelial Cell Capillary Formation
Authors: Rigassi Lisa , Barchiesi Federica , Unterleutner Elisabeth , Imthurn Bruno , Dubey Raghvendra K
Published: March 2022

MicroRNAs play a key role in vascular remodeling associated with cardiovascular disease. MiR-221 actively contributes to injury-induced neointima formation by inhibiting endothelial cell (EC) growth and promoting smooth muscle cell (SMC) growth. Since estradiol (E2) prevents neointimal thickening, we hypothesize that E2 mediates its vasoprotective actions by downregulating miR-221 expression and abrogating its effects on SMC and EC growth. RT-qPCR confirmed that both Human Umbilical Vein ECs (HUVECs) and Human Coronary Artery SMCs (HCASMCs) produce miR-221. Treatment of HCASMCs with PDGF-BB (20ng/ml) induced miR-221 levels from 100±8% to 189±9% (p<.05) and treatment with E2 (100nM) inhibited this to 126±4% (p<.05 vs PDGF). PDGF-BB stimulated DNA synthesis (BrdU incorporation), CyclinD1 expression (Western Blot) and migration (Scratch assay) in HCASMCs and these effects were mimicked by miR-221 (25nM) overexpression; and abrogated in HCASMCs transfected with miR-221 antimiR (25nM). E2 inhibited PDGF-induced HCASMC numbers by 30±4% and these effects were reversed by miR-221 (p<0.05). Inhibitory effects of E2 on PDGF-induced miR-221 production in HCASMCs were mimicked by estrogen receptor (ER) α agonist PPT, but not by ERβ and GPER agonists; and blocked by ERα antagonist MPP, suggesting these effects are ERα mediated. In contrast to SMCs, transfection of HUVECs with miR-221 mimic inhibited capillary formation and wound healing by 39±8% and 27±6%, respectively (p<.05). Neutralization of miR-221 with antimiR induced capillary formation and wound closure by 26±3 % and 51±15%, respectively (p<.05). E2 (10nM) inhibited miR-221 levels in HUVECs from 100 to 73±6% (p<.05). Moreover, transfection of HUVECs with miR-221 mimic inhibited E2-induced capillary formation (from137±9% to 85±13%; p<.05 vs E2) and wound closure (from 125±5% to 82±12%; p<.05 vs E2). Our findings provide the first evidence that E2 inhibits miR-221 production in HCASMCs and HUVECs and these effects contribute to its antimitogenic effects on HCASMCs and capillary promoting effects in HUVECs. Modulation of miR-221 by E2 represents a novel mechanism by which E2 may mediate its differential effects on SMC and EC growth, and confer vascular protection.
Abstract P045: High Methionine, Low Folate and Vitamin B6/B12 Containing Diet Leads to Cognitive Functions Impairment by Epigenetic Silencing of Netrin Gene
Authors: Kalani Anuradha , Nuru Mohammed , Kamat Pradip K , Lominadze David , Tyagi Neetu
Published: March 2022

Hypermethylation of the genes silence the transcriptions of functional genes associated to cognitive functions. Loss of memory due to epigenetic modifications, called memory-epigenetics was studied. Netrin is a glycoprotein involved in neurogenesis, axonal guidance and maintenance of synaptic plasticity. We hypothesized that high methionine-low vitamins containing diet leads to memory impairment by increasing global DNA methylation and therefore, silencing the netrin gene. Wild type (C57BL/6J) mice were fed diet containing excess met (1.2%), low-folate (0.08 mg/kg), vitamin B6 (0.01 mg/kg), and B12 (10.4 μg/kg) for 6 weeks. Mice were weekly examined for cognitive functions. Our results using a passive avoidance test confirmed a loss in fear motivated long-term memory starting from 4th week that continued up to 6th week of diet (F=37.14, p<0.001). The loss of memory was negatively associated with an increase in global DNA methylation in mice brains fed with diet (R2=-0.99, p<0.001). We found a time-dependent decrease in netrin protein expression (F=27.63, p<0.001) and an increase in methylation of netrin gene promoter, defined by restriction digestion-PCR analysis, in mice fed with diet. The increase in methylation of netrin gene promoter was further validated by high resolution melting and sequencing analysis. In addition, the association of netrin with memory impairment was confirmed by delivering netrin protein to diet-fed mice brains through intracerebral route. The data suggest that netrin introduction helped in considerable memory regain (~50%) in mice on diet. Taken together, these results suggest that high met, low folate and vitamin B6/B12 containing diet can induce defects in learning and memory. Furthermore, the data indicates that decrease in netrin level due to hypermethylation of its gene promoter can be associated with memory loss.
Abstract P047: Epigenetic Mechanisms of Sodium Butyrate- and Retinoic Acid-dependent Attenuation of Renal Fibrosis and Inflammation in Npr1 Gene-targeted Mice
Authors: Kumar Prerna , Pandey Kailash N
Published: March 2022

The objective of the present study was to determine the combined effect of sodium butyrate (NaBu), a histone deacetylase (HDAC) inhibitor and all-trans retinoic acid (ATRA) on attenuation of renal fibrosis and inflammation in Npr1 (coding for natriuretic peptide receptor-A) gene-disrupted mutant mice. Adult (18-20 week old) male Npr1 gene-disrupted heterozygous (1-copy; Npr1+/-) wild-type (2-copy; Npr1+/+), and gene-duplicated (3-copy; Npr1++/+) mice were treated by injecting ATRA-NaBu hybrid drug (1.0 mg/kg/day) intraperitoneally for 2-weeks. A marked attenuation in tubulo-interstitial fibrosis was observed in Npr1+/- mice after treatment with ATRA-NaBu (50%, p < 0.001). Western blot analyses exhibited the reduction in renal expression of collagen type I alpha 2 and transforming growth factor-beta (55%, p < 0.001, 67%, p < 0.001, respectively) in ATRA-NaBu-treated Npr1+/- mice compared with vehicle-treated mice. A significant decrease in systolic blood pressure was observed in ATRA-NaBu-treated Npr1+/- mice (treated 110.3 ± 2.6 vs. Npr1 +/- control mice, 126.3 ± 2.7, p < 0.01). The ATRA-NaBu also enhanced plasma cGMP levels (pmol/ml) in Npr1+/- (treated, 22.7 ± 3.3 vs. control, 6.2 ± 1.6; p < 0.05), Npr1+/+ (treated, 50.9 ± 3.5 vs. control, 20.8 ± 3.6; p < 0.05), and Npr1++/+ (treated, 71.4 ± 6.4 vs. control, 34.5 ± 2.9; p < 0.05) mice. Treatment with ATRA-NaBu significantly lowered renal levels (pg/mg protein) of monocyte chemoattractant protein-1 (MCP-1) (treated, 109.5 ± 9.9 vs. control, 24.3 ± 1.7; p < 0.01) in Npr1+/- mice compared with vehicle-treated mice. Western blot analyses confirmed the reduction in renal expression of MCP-1 (72%, p < 0.001) in ATRA-NaBu-treated Npr1+/- mice compared with control mice. Moreover, the increased HDAC activity in Npr1+/- mice was significantly reduced by ATRA-NaBu treatment compared with untreated Npr1+/- control mice. The present results provide direct evidence that ATRA-NaBu acts as a potent antifibrotic agent and repairs the renal pathology in Npr1+/- mice, which will have important implications in prevention of hypertension-related renal pathophysiological conditions.
Abstract P049: Relationship between Serum Ghrelin levels and Primary Hypertension: A Meta Analysis
Authors: Agasthi Pradyumna , Aloor Sivakanth , Chenna Avantika , Chinta Siddharth R , Onwuanyi Anekwe
Published: March 2022

Background:
Ghrelin is a novel gut derived peptide hormone suggested to play a role in the etiopathogenesis of Primary Hypertension (HTN). It is hypothesized that ghrelin regulates arterial blood pressure via modulation of central sympathetic activity and peripheral nitric oxide dependent vasodilatory mechanisms. The aim of the present study is to conduct a meta-analysis to evaluate the relationship between serum ghrelin levels and HTN.

Methods:
We searched MEDLINE, CINHAL and COCHRANE databases for studies reporting serum ghrelin levels in the HTN and non HTN study population. We included case controls, cohort and cross-sectional studies. We calculated the weighted standardized mean difference (SMD) in serum ghrelin levels between the HTN and control groups.

Results:
Our search strategy yielded 309 articles and we included 8 studies enrolling 1659 participants. The median age of the HTN group was 52 yrs. (IQR 47-56) compared to 49 yrs.(IQR 39-54) in the control group. The median body mass index in the HTN group was 28 kg/m2 (IQR 27-34) compared to 25 kg/m2 (IQR 23-27) in the control group. The median percentage of female population in the HTN group was 48 % (IQR 37-61) compared to 51 %(IQR 32-57) in the control group. The unweighted median serum ghrelin levels in the HTN group were 733 pg/ml (IQR 571-5607) compared to 857 pg/ml (IQR 592-7387) in the control group. The SMD of serum ghrelin level was -1.381 (95% CI -2.089,-0.673) p<0.001 comparing those in the HTN group and control group.

Conclusion:
Serum ghrelin levels are significantly and negatively associated with HTN. Further studies are needed to address potential confounding factors and confirm this association.
Abstract P054: Inhibition of Endosomal MyD88-dependent Signaling with Chloroquine Improves Endothelial Function and Lowers Blood Pressure in Spontaneously Hypertensive Rats
Authors: McCarthy Cameron G , Wenceslau Camilla F , Goulopoulou Styliani , Ogbi Safia , Baban Babak , Matsumoto Takayuki , Webb R. Clinton
Published: March 2022

Uncontrolled inflammation and chronic immune system activation are common in hypertension. However, the exact mechanisms by which they occur are not well understood. Innate immune system recognition and response to damage-associated molecular patterns (DAMPs) is becoming an increasingly accepted mechanism. Mitochondrial DNA (mtDNA) is a DAMP that is recognized by Toll-like receptor (TLR)9 and is elevated in the circulation of spontaneously hypertensive rats (SHR). Therefore, we sought to determine the contribution of TLR9 in hypertension. We hypothesized that TLR9 inhibition with lysosomotropic agent chloroquine would improve endothelial function and lower blood pressure in SHR. We treated adult SHR and control Wistar-Kyoto rats (12 weeks old), as well as a group of young SHR (5 weeks old) with chloroquine (40 mg/kg/day, i.p.) for 21 days. Chloroquine lowered adult SHR blood pressure (Veh: 201±2 vs. CQ: 182±5 mmHg, p<0.05) and inhibited cyclooxygenase-dependent contraction to acetylcholine (ACh) in isolated mesenteric resistance arteries (MRA) from adult SHR [%NE (ACh 1 μmol/L), Veh: -31±8 vs. CQ: 70±10, p<0.05]. Chloroquine also inhibited co-localization of not only TLR9, but also TLR7 and TLR8 (MyD88-dependent endolysosomotropic TLRs) with MyD88 in adult SHR MRA, as well as expression of MyD88 in young SHR MRA. Finally, we observed that prophylactic treatment with chloroquine, during the critical pre-hypertensive stage, ameliorated the development of high blood pressure (Veh: 190±4 vs. CQ: 174±5 mmHg, p<0.05) and components of the adaptive immune system (decreased aortic-derived CD45+ leukocytes, circulating CD3+ T lymphocytes, and expression of CD44 on circulating and aortic-derived CD3+ T lymphocytes) upon maturation to adulthood in SHR. In conclusion, endosomal MyD88-dependent signaling from TLR7, 8, and 9 contribute to high blood pressure, endothelial dysfunction, and recruitment of the adaptive immune system in SHR. These findings support the involvement of the innate immune system in the pathogenesis and maintenance of hypertension.
Abstract P062: Long-term Exposure to Endothelin-1 Overexpression Increases Blood Pressure and Causes Small Artery Injury
Authors: Coelho Suellen C , Ouerd Sofiane , Fraulob-Aquino Júlio C , Offermanns Stefan , Paradis Pierre , Schiffrin Ernesto L
Published: March 2022

Background:
The mechanisms of blood pressure (BP) regulation by endothelin (ET)-1 produced by endothelial cells are complex and remain unclear. Recently, we developed a transgenic mouse with tamoxifen-inducible endothelium-restricted human ET-1 overexpression (ieET-1) using Cre/loxP technology. ieET-1 mice exhibited BP rise after three weeks of induction in an ET type A receptor-dependent manner, in absence of vascular and kidney injury. It is unknown whether long-term exposure to ET-1 overexpression results in elevated BP elevation and vascular injury.

Methods:
Nine to 12-week old male ieET-1 mice and control ieCre mice expressing a tamoxifen-inducible Cre recombinase (CreERT2) under the control of EC-specific Tie2 promoter, were treated with tamoxifen (1 mg/kg/day, s.c.) for 5 days and studied 3 months later. Metabolic cages were used to collect 24-hour urine for sodium, potassium and protein measurements. Renal artery flow (RAF) was assessed by ultrasonography. BP was determined by telemetry, plasma aldosterone by ELISA, and small mesenteric artery (MA) endothelial function and vascular remodeling by pressurized myography.

Results:
Systolic BP was increased in ieET-1 compared with ieCre mice (144±5 vs 117±3 mmHg, P<0.001). RAF was decreased in ieET-1 compared with control (1.9±0.2 vs 3.0±0.3 mL/min, P<0.01). The excretion of urinary sodium, potassium and protein was similar in both groups. Plasma aldosterone levels were increased in ieET-1 compared with ieCre mice (1.99±0.20 vs 1.29±0.12 ng/mL, P<0.05). Endothelium-dependent relaxation responses to acetylcholine were impaired in ieET-1 compared to ieCre mice (36.3±4.7 vs 71.4±9.7%, P<0.01), whereas endothelium-independent relaxation responses to sodium nitroprusside were unchanged. MA media/lumen and media cross-sectional area were similar in both groups, but stiffness was increased in ieET-1 compared to ieCre mice, as indicated by leftward displacement of the stress-strain curves (strain at 140mmHg: 0.61±0.04 vs 0.71±0.02, P<0.05).

Conclusions:
The results demonstrate that long-term exposure to endothelial ET-1 overexpression caused sustained BP rise and small artery stiffening.
Abstract P065: Pacemaker Tapping Maneuver in the Treatment of Orthostatic Hypotension
Authors: Ashangari Chandralekha , Suleman Amer
Published: March 2022

A 77 year old man was referred to us with fainting spells especially after standing/walking few steps. Initially he went to a neurologist and ENT specialist where the cause could not be determined and symptoms continued. During annual physical, he noticed that his blood pressue dropped when he stood up suddenly. He had severe fainting spells and was placed pacemaker in him.However,pacemaker did not make a huge difference.Blood Pressure monitoring was done and noticed 50 mmHg drop upon standing. Pacemaker was changed to constant pacing where his dizziness continued but he had no falls. He had periodic vision problems, numbness in fingers and legs, digestive problems, stomach issues and balance problems. His tilt table test showed orthostatic hypotension.

Medical management (florinef, mestinone, midodrine) did not relieve his symptoms .Pacemaker sensor was turned on and made more sensitive that after he taps on the pacemaker, heart rate increases for 10 minutes. We told him to tap on pacemaker whenever he is dizzy and see if there is any improvement in orthostatic symptoms. The later treatment had made a huge difference; he is practically asymptomatic and was very much satisfied with his current health. He is using the tapping maneuver 4-5 times a day and his fainting disappeared and no syncope.

First time our case report on Pacemaker tapping demonstrated significant improvement in treating symptoms of orthostatic hypotension.
Abstract P068: Comparison Effects of Leptin and Phentolamine on Cardiovascular System
Authors: Hasan Khaled M
Published: March 2022

Background:
Leptin participates not only in negative feedback control of body adiposity but in cardiovascular and sympathetic regulations. The aim of this research is to study the acute effects of leptin on cardiovascular system. Rabbits were used as experimental animals.

Materials and Methods:
Blood pressure and EKG have been recorded at basal and after injection of these medications. Intravenous bolus of 500μg/kg of leptin has been injected into marginal vein in first group (15 rabbits), whereas second group (15 rabbits), has been injected α-adrenergic receptor antagonist phentolamine (0.01 mg/kg).

Results:
First group of animals received IV leptin 500 microgram/kg, there was significant increase in mean arterial blood pressure from 75.60±0.61 mmHg to 94.63±0.51 mmHg (P<0.001). Moreover, heart rate has been increased after IV leptin, from 225±3.04 beat/min to 291±4.42 beat/min (P<0.001).

Second group of animals received phentolamine significantly decreased response of cardiovascular action of leptin. Leptin increased blood pressure nonsignificantly. Mean arterial blood pressure was before treatment 74.70±0.51 mmHg and became after treatment with phentolamine and leptin in sequences 77.63±0.41 mmHg (P>0.05). However, phentolamine did not impair effects of leptin on heart rate. Heart rate was 229±2.41 beat/min and significantly increased after treated with phentolamine and leptin in sequences 344±3.52 beat/min (P<0.001). This study elucidated significant positive relationship between blood pressure and heart rate in group which is treated with leptin (p<0.001), but this relationship was inversely significant related in group two which is treated with phentolamine (p<0.001).

Conclusions:
These results suggest that intravenous injection of leptin causes increases in arterial blood pressure and heart rate in anesthetic rabbits, and these effects could be opposed by adrenergic blockade. This may be partially explained by the effect of leptin on cardiovascular system could be mediated by sympathetic pathway.
Abstract P072: Adipocyte AT
2
Receptor Suppresses UCP1 Transcription and Thereby Resting Metabolic Rate
Authors: Littlejohn Nicole K , Weidemann Benjamin J , Pearson Nicole A , Naber Meghan C , Potthoff Matthew J , Klingelhutz Aloysius , Sigmund Curt D , Grobe Justin L
Published: March 2022

The renin-angiotensin system (RAS) contributes to energy balance through opposing actions in the brain and periphery. We hypothesize that tissue- and receptor-specific RAS modulation may represent a novel therapeutic approach to obesity. Transgenic “sRA” mice exhibit brain-specific RAS hyperactivity through expression of human renin in neurons (synapsin promoter) and human angiotensinogen via its own promoter. Previously we documented that sRA mice exhibit a suppressed circulating RAS, and an elevated resting metabolic rate (RMR) that is sensitive to replacement of circulating angiotensin II or the AT
2
receptor (AT
2
R) agonist, CGP-42112a (CGP, 100 ng/kg/min, s.c.). sRA mice exhibit a robust and specific elevation in uncoupling protein-1 (UCP1) expression and glucose uptake in inguinal white adipose tissue (iWAT). Chronic infusion of the AT
2
R agonist, CGP-42112a (CGP) into sRA mice increased weight gain and normalized RMR and inguinal fat expression of UCP1, but had no effect on food intake or digestive efficiency. RMR suppression by CGP was not mediated through attenuation of adipose sympathetic nervous activity (SNA, measured via multifiber nerve recordings), leading to the hypothesis that AT
2
R action on the adipocyte suppresses adipose sensitivity to SNA. Cultured primary mouse adipocytes were isolated from the iWAT, and norepinephrine (NE, 1 nM) treatment induced UCP1 mRNA (597 -fold of vehicle, P<0.05), however this UCP1 induction was attenuated with CGP co-treatment (10 nM, 0.2 -fold of NE alone, P<0.05). In human adipocytes, activation of AT
2
R also prevented the NE-induced increase in UCP1 mRNA. NE additionally significantly increased lipolysis by adipocytes (measured via free glycerol in the media; P<0.05), but AT
2
R activation had no effect (vehicle= 15.9 ug/mL; NE= 70.8 ug/mL; NE + CGP=64.7 ug/mL). These data support a suppressive action of AT
2
R upon RMR that is mediated specifically through the suppression of UCP1 expression, but not its activation by lipolysis, within subcutaneous adipocytes. Ongoing studies are examining the transcriptional regulation of UCP1 by adipose AT
2
R activation.
Abstract P077: Atorvastatin Ameliorates Cardiac Injury and Inflammation via Adiponectin-independent Activation of AMP-activated Protein Kinase and Inhibition of the Nf-κB Pathway in Rats With Metabolic Syndrome
Authors: Yamada Yuichiro , Takeuchi Shino , Watanabe Shogo , Minagawa Yuji , Ito Shogo , Sano Yusuke , Nagasawa Kai , Matsuura Natsumi , Murohara Toyoaki , Nagata Kohzo
Published: March 2022

Background:
Statins have been implicated in inhibition of inflammation and preventing new onset type 2 diabetes. However, the mechanism underlying the pleiotropic effects of statins remains controversial. We investigated the effects of atorvastatin on cardiac and adipose tissue pathology as well as glucose and lipid metabolism in rats with metabolic syndrome (MetS).

Methods and Results:
We used DahlS.Z-Leprfa/Leprfa (DS/obese) rats, derived from a cross between Dahl salt-sensitive and Zucker rats, as a new animal model of MetS. DS/obese rats were treated with atorvastatin (6 or 20 mg/kg/day, p.o.) from 9 to 13 weeks of age. Treatment with atorvastatin ameliorated LV fibrosis and diastolic dysfunction as well as LV oxidative stress and inflammation without affecting hypertension or LV hypertrophy in DS/obese rats. Atorvastatin did not affect visceral or subcutaneous fat mass but alleviated adipocyte hypertrophy (high-dose) and inflammation (both high- and low-dose) in visceral adipose tissue in these rats. Both high- and low-dose atorvastatin reduced serum adiponectin concentration in DS/obese rats. However, both doses of atorvastatin similarly attenuated the decrease in phosphorylation of AMP-activated protein kinase (AMPK) as well as the increase in phosphorylation of NF-kB in the heart. Insulin resistance was similarly improved by both high- and low-dose atorvastatin. Glucose intolerance was partially ameliorated by low-dose atorvastatin and completely prevented by high-dose atorvastatin. Both doses of atorvastatin similarly attenuated the increases in serum levels of total cholesterol, LDL-cholesterol, and triglycerides in DS/obese rats.

Conclusions:
Treatment of DS/obese rats with atorvastatin attenuated adipose tissue inflammation, without affecting obesity, as well as ameliorated LV inflammation, fibrosis and diastolic dysfunction and abnormal metabolism. The beneficial cardiac effects of atorvastatin are likely attributable, at least in part, to adiponectin-independent activation of AMPK and subsequent inhibition of the NF-κB pathway.
Abstract P083: Gender Specific Effects in Nox4-/- Mice in Hypoxia Induced Pulmonary Hypertension and Pulmonary Vascular Resistance
Authors: Hood Katie Y , Harvey Adam P , Nilsen Margaret , Montezano Augusto C , Touyz Rhian M , MacLean Margaret R
Published: March 2022

Oxidative stress and Noxs have been implicated in the pathogenesis of experimental and human pulmonary arterial hypertension (PH). Gender differences in PH may be, in part, due to increased formation of NADPH oxidase (Nox) derived reactive oxygen species (ROS). A large body of evidence implicates E2 in the pathogenesis of PAH and an interaction between E2 and Noxs has been suggested. We hypothesised that i) Estrogen (E2) leads to Nox-induced oxidative stress, which promotes PASMC damage, ii) E2-induced Nox activation may account for gender differences in PH.

Cultured human PASMC were stimulated with E2 (1nM). ROS production was assessed by chemiluminescence (O2-) and proliferation by BrdU assay. E2 increased superoxide (219%; p<0.05) and proliferation in PASMC, effects blocked by GKT137831 (Nox4/1 inhibitor).

In vivo studies were conducted to assess the role of Nox4 in hypoxia-induced pulmonary hypertension in male and female mice. Hypobaric hypoxia was used to induce PH in mice, which were divided into 8 groups: normoxic WT (NWT), hypoxic WT (HWT), normoxic Nox4-/- (NKO) and hypoxic Nox4-/- (HKO) for both male and female mice. In male HWT mice, RVSP (20.5 NWT vs. 45.2mmHg HWT, p<0.001) and RVH by Fulton Index (0.20 NWT vs. 0.395 HWT, p<0.01) were increased, an effect that was significantly reduced in male Nox4-/- mice (RVSP: 36.4mmHg HKO; RVH: 0.030 HKO; p<0.05). In female HWT mice, RVSP (21.4 NWT vs. 39.8mmHg HWT, p<0.05) and RVH (0.20 NWT vs. 0.31 HWT, p<0.01) were elevated in hypoxia, yet female Nox4-/- mice were not protected against hypoxia-induced PAH (RVSP: 34.2mmHg HKO; RVH: 0.33 HKO).

Hypoxia-induced endothelial dysfunction in both male and female WT pulmonary arteries was improved in male HKO, however, endothelial dysfunction remained in HKO females. In female Nox4-/- mice, increased spleen and uterine weight (which has been associated with altered ovarian hormone biogenesis) suggests a role for inflammatory and fibrotic processes.

In conclusion, genetic ablation of Nox4 in males, but not females, protects against the development of hypoxic PH. The effects of E2 on oxidative stress is present in PASMCs where Nox4-derived ROS may be an important regulator of, and impact on, molecular processes contributing to vascular remodelling in PAH.
Abstract P086: Superoxide but Not Hydrogen Peroxide Increases Nuclear Translocation of Transcription Factor Sp3 and AT1 Receptor Expression in the Renal Cells
Authors: Saleem Mohammad
Published: March 2022

Age-associated oxidative stress causes up-regulation of renal AT1 receptor function (AT1R) and hypertension in aging Fischer Brown Norway (FBN) rats. Here we studied the mechanism of up-regulation of renal AT1R, and further tested superoxide Vs hydrogen peroxide (H2O2) specificity in this phenomenon. We found that transcription factor Sp3 plasmid increased (Control vs Sp3: 0.1165 ± 0.01 vs 0.3810 ± 0.03) while Sp3 siRNA decreased (Control siRNA vs Sp3 siRNA: 1.11 ± 0.25 vs 0.64 ± 0.06) the levels of AT1 receptor protein in human kidney (HK2) cells. Whereas transcription factor NF-kB p-65 plasmid did not affect AT1 receptor protein levels in these cells (Control vs NF-kB: 0.25 ± 0.025 vs 0.31 ± 0.035). DDC, a superoxide prodrug, but not H
2
O
2
treatments increased nuclear levels of Sp3 and NF-kB proteins in HK2 cells [(Control vs DDC, vs H
2
O
2,
vs H
2
O
2
+ tempol): Sp3 (0.50 ± 0.08 vs 1.28 ± 0.21 vs 0.68 ± 0.14 vs 1.04 ± 0.30 densities); NF-kB (0.4025 ± 0.13 vs 1.808 ± 0.54 vs 0.4950 ± 0.15 vs 0.6375 vs ± 0.18 densities)]. Tempol treatment, a superoxide scavenger, attenuated DDC-mediated nuclear accumulation of Sp3 (DDC vs DDC + tempol: 1.28 ± 0.21 vs 0.52 ± 0.12 densities) and NF-kB (DDC vs DDC + tempol: 1.808 ± .54 vs 0.43 ± 0.18 densities]. In addition, DDC but not H
2
O
2
increased AT1 receptor mRNA expression, measured by RT-qPCR [(Control vs DDC, vs H
2
O
2
): (1.000 ± 0.0 vs 3.323 ± 0.79, vs 1.218

± 0.49)]. This effect was attenuated by tempol treatment [(DDC vs DDC + tempol) (3.323 ± 0.79 vs 0.7225 ± 0.16 densities)]. Similarly, DDC treatment increased AT1 receptor protein expression that was attenuated by tempol treatment, measured by immunoblotting [(Control vs DDC vs DDC + Tempol) (0.3535 ± 0.99 vs 0.9975 ± 0.19 vs 0.3250 ± 0.02 densities)]. These results suggest that superoxide but not H
2
O
2
regulates both Sp-3 and NF-kB in the renal cells. However, it is Sp3 but not NF-kB that up-regulates renal AT1 receptor expression. Taken together, this phenomenon may represent a mechanism for hypertension in aging FBNs.
Abstract P087: PDI via Keap1/Nrf2 Pathway Regulates Renal AT1 Receptors and Blood Pressure in Aging Rats
Authors: Wang Xitao , Asghar Mohammad
Published: March 2022

Renal angiotensin AT1 receptor (AT1R) plays pivotal role in the regulation of blood pressure (BP). We recently reported that age-associated oxidative stress causes hyper-activation of AT1R and hypertension in the aging Fischer Brown Norway (FBN) rats. Our objective was to understand the mechanism of higher oxidative stress contributing to the impaired AT1R function in the aged FBNs. Nrf2 transcription factor protects kidney from oxidative stress by activating the transcription of diverse antioxidant and detoxifying enzymes. Kelch-like ECH-associated protein 1 (keap1) inhibit Nrf2 activity by sequestering Nrf2 in cytosol. We hypothesize that protein disulfide isomerase (PDI), an enzyme involved in isomerization reaction, regulates keap1 and Nrf2. Nrf2, keap1 and PDI were determined by Western blotting and/or immunohistochemistry in the kidney of FBNs and human kidney 2 (HK2) cells. We found that the aging kidney had low levels of PDI (adult vs old: 5.38±2.60 vs 3.59±1.76, 33.3% decrease), which was associated with low nuclear levels of Nrf2 (adult vs old: 1.86±0.69 vs 0.87±0.18, 53.2% decrease) and high cytosolic levels of keap1 (adult vs old: 1.80±0.46 vs 2.40±0.49, 33.3% increase) in the aging FBNs. Moreover, PDI inhibition by bacitracin in HK2 cells significantly reduced Nrf2 accumulation (control vs treated: 1.25±0.26 vs 0.50±0.14, 60% decrease) in the nuclei. PDI siRNA treatment in HK2 cells decreased the level of PDI (scrambled vs PDI siRNA: 0.64±0.07 vs 0.52±0.08) while increased the level of AT1R (scramble vs PDI siRNA: 0.11 vs 0.20). These data suggest that PDI by regulating keap1/Nrf2 redox-pathway regulates AT1R, and may be involved in BP regulation in the aging FBNs.
Abstract P088: Pravastatin Protects Against Glucose-induced Anti-proliferative, Anti-invasive and Anti-angiogenic Milieu in Cytotrophoblasts
Authors: Afroze Syeda H , Wesley Katy B , Drever Nathan , Costantine Maged M , Allen Steven R , Zawieja David C , Kuehl Thomas J , Uddin Mohammad N
Published: March 2022

Objective:
An increasing level of evidence supports the utility of pravastatin in prevention against preeclampsia (preE). We previously demonstrated that hyperglycemia induces cytotrophoblasts (CTBs) dysfunction characteristic of a preE-like phenotype. We sought to demonstrate the utility of pravastatin in rescuing CTBs from hyperglycemia induced dysfunction.

Methods:
Human CTBs were treated with 100, 150, 200, 300, or 400 mg/dL glucose for 48h. Some cells were pretreated with pravastatin (1ug/mL) for 2h, while others were co-treated with pravastatin (1ug/mL) prior to glucose treatment. Some cells were treated with D-Mannitol. Cell migration was performed by Matrigel migration assay kit according to manufacturer protocol. Cell lysates were utilized to evaluate the expression of urokinase plasminogen activator (uPA), plasminogen activator inhibitor 1 (PAI-1), proliferating cell nuclear antigen (PCNA) and p38 MAPK phosphorylation by western blot. Levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin (sEng) and interleukin 6 (IL-6) were measured in culture media using ELISA kits. Statistical comparisons were performed using analysis of variance with Duncan’s post hoc test.

Results:
Hyperglycemia inhibited CTBs migration, down-regulated uPA, PAI-1, PCNA and up-regulated p38 phosphorylation in CTBs treated with ≥150 mg/dL glucose compared to basal (100 mg/dL) (*p < 0.05 for each). Secretion of sFlt-1, sEng and IL-6 were increased while VEGF and PIGF were decreased in CTBs treated ≥150 mg/dl of glucose (*p < 0.05 for each). Both pravastatin pretreatment and co-treatment significantly rescued CTBs migration, up-regulated uPA, PAI-1, PCNA, down-regulated p38 phosphorylation, and corrected the angiogenic profile of CTBs (p < 0.05 for each). D-Mannitol had no effect on CTBs.

Conclusions:
Pravastatin mitigates the hyperglycemia-induced dysfunction of CTBs by attenuating the glucose-induced anti-proliferative, anti-invasive and anti-angiogenic phenotype. These data should alleviate critical concerns regarding pravastatin use on CTBs development early in pregnancy and support the current research to use of pravastatin in preE prevention.
Abstract P090: Agonistic Autoantibodies to the Angiotensin II Type 1 Receptor Enhances ANG II Induced Renal Vascular Sensitivity and Reduces Renal Function During Pregnancy
Authors: Cunningham Mark W , Williams Jan , Wallukat Gerd , Dechend Ralf , LaMarca Babbette
Published: March 2022

Preeclamptic women produce agonistic autoantibodies to the Angiotensin II type 1 receptor (AT1-AA) and exhibit increased blood pressure (BP) and vascular sensitivity to angiotensin II (ANG II). Although, together AT1-AAs and ANGII increase the BP, renal artery resistant index, and vasoconstriction of renal afferent arterioles in pregnant rats; the renal hemodynamics in the presence of the AT1-AAs during pregnancy has not been examined. Thus the objective of this study was to examine the changes in the glomerular filtration rate (GFR) and renal blood flow (RBF) during pregnancy in the presence of AT1-AAs and/or ANGII. Methods: Pregnant Sprague Dawley rats were divided into 4 groups: Normal Pregnant (NP, n=6), Pregnant + ANG II (Preg + ANG II, n=6), Pregnant + AT1-AA (Preg + AT1-AA, n=8), and Pregnant + ANG II + AT1-AA (Preg + ANGII + AT1-AA, n= 6). On day 13 of pregnancy, rats were implanted with mini-pumps infusing ANG II (50 ng/kg/min) and/or AT1-AA (1:40 dilution). On day 19 of pregnancy, rats were subjected to terminal renal function surgeries using FITC labeled Inulin. During the surgery, the BP was recorded and a transonic flowmeter probe was placed on the left renal artery to measure RBF. Results: BP was elevated in all pregnant rats administered ANG II and/or the AT1-AA. Although GFR was reduced, it was not significant between Preg + ANG II and Preg + AT1-AA vs. NP rats (1.5 ±0.24, 1.60 ±0.17 vs. 1.90 ±0.16 ml/min). However, the GFR was further decreased in Preg + ANGII + AT1-AA rats (1.20 ±0.08). No difference was observed with the RBF between Preg + ANG II and Preg + AT1-AA vs NP rats (14.4 ±2.96, 14.4 ±1.48 vs. 15.4 ±1.75 ml/min). RBF was decreased in Preg + ANGII + AT1-AA vs NP rats (7.4 ±1.09 vs. 15.4 ±1.75 ml/min). No change in RVR between Preg + ANG II and Preg + AT1-AA vs. NP rats (9.7 ±2.69, 8.3 ±0.58 vs. 6.4 ±0.77). However, the RVR was drastically increased between Preg + ANGII + AT1-AA vs NP rats (18.4 ±2.91 vs. 6.4 ±0.77). Conclusion: Together ANG II and AT1-AA drastically decreases renal function by 37%, RBF by 50%, and caused a 3 fold increase in RVR vs NP rats. These data indicate the importance of AT1-AAs to drastically enhance ANG II induced renal vascular sensitivity and reduce renal function during preeclampsia. Research Supported by T32HL105324 and RO1HD067541