bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P093: The Sflt-1/plgf Ratio Associates with Prolongation of Pregnancy
Authors: Saleh Langeza , Verdonk Koen , Danser Jan A , Steegers Eric A , Russcher Henk , van den Meiracker Anton H , Visser Willy
Published: March 2022

Objective:
To evaluate whether a single determination of the serum sFlt-1/PlGF ratio associates with pregnancy prolongation in women with suspected or confirmed preeclampsia (PE).

Methods:
In this ongoing observational Dutch multicenter study (600 pts to be enrolled) blood was drawn at admission. Values of sFlt-1 and PlGF were measured postpartum using the automated Elecsys system to prevent influence of this information on decision-making of the treating physicians and the defining time point of delivery. Clinical characteristics and pregnancy outcomes were retrieved from medical records. Cutoffs of < 33 to rule out and > 85 to rule in the occurrence of delivery for selected time points were used.

Results:
We have included 412 patients, so far complete data of 147 patients (age 18 to 48 yrs. singleton pregnancies, median pregnancy duration 32 weeks (range 20-36 weeks) is available. At time of inclusion 67 pregnancies were complicated with PE, of which 12 with superimposed PE, 8 with HELLP, 20 with gestational hypertension (GH) and 60 without pregnancy induced hypertension (PIH). The median (range) ratio in patients who delivered within 7 days was 146 (9-1803) compared to 14 (1-469) in those who delivered after > 7 days (p< 0.001). Delivery within 7 days occurred in 59% patients with a ratio > 85 compared to 7% with a ratio <33 (p=0.002) and 39% with a ratio 33-85 (p=0.048).

Conclusion:
In this high risk group a low ratio is inversely correlated with prolongation of pregnancy.
Abstract P095: Effects of Obesity on Placental Ischemia-induced Hypertension
Authors: Spradley Frank T , Palei Ana C , Granger Joey P
Published: March 2022

Although it is known that obesity is a risk factor for preeclampsia (PE), the mechanisms are not clear. Placental ischemia stimulates the release of the antiangiogenic factor sFlt-1 into the maternal circulation eliciting vascular dysfunction and hypertension. Therefore, we tested the hypothesis that placental ischemia (reduced uterine perfusion pressure, RUPP)-induced hypertension and sFlt-1 levels are exaggerated in obese rats. MC4R-deficient obese rats (MC4R+/-) and wild-type Wistar Hannover controls (MC4R+/+) were maintained on NIH31 standard chow; mated at 17 weeks old; RUPP surgeries performed at gestational day (GD) 14; and mean arterial blood pressure (MAP) and pregnancy weights assessed at GD 19. This resulted in 4 groups: normal pregnant (NP) MC4R+/+ (N=10), RUPP MC4R+/+ (N=12), NP MC4R+/- (N=12) and RUPP MC4R+/- (N=11). Body weight was greater in NP MC4R+/- than NP MC4R+/+ (371±10 vs. 340±6, P<0.05), which was reduced by RUPP in both rat strains but MC4R+/- were still heavier (327±13 vs. 297±8, P<0.05). Total body fat mass by EchoMRI was greater in NP MC4R+/- than MC4R+/+ (72±6 vs. 45±4, P<0.05) whereas fat mass was not altered in RUPP MC4R+/+ (38±4g) but was reduced in RUPP MC4R+/- (56±5) (P<0.05). Fetal weights were similar between NP MC4R+/+ (1.93±0.04) and MC4R+/- (1.95±0.02), which was reduced to a greater extent in RUPP MC4R+/- than MC4R+/+ (1.62±0.03 vs. 1.74±0.03, P<0.05). MAP was slightly elevated in NP MC4R+/- over MC4R+/+ (107±2 vs. 101±1). RUPP significantly increased MAP in MC4R+/+ (117±2, P<0.05) but not MC4R+/- (113±3). Plasma leptin levels were greater in NP MC4R+/- over MC4R+/- (6.7±1.0 vs. 3.6±0.4, P<0.05) whereas RUPP had no effect on these levels in MC4R+/+ (3.8±0.7) or MC4R+/- (6.5±1.1). Plasma and placental levels of sFlt-1 were similar in all groups. Clinically, not all PE women have elevated sFlt-1 levels. Contrary to our hypothesis, MAP was not exaggerated by placental ischemia in obese rats having MC4R deficiency. In fact, MAP was not significantly increased following placental ischemia in MC4R-deficient rats indicating that intact MC4R signaling, which has been shown to promote the development of hypertension in several animal models by sympathetic mechanisms, mediates the development of PE independent of sFlt-1.
Abstract P102: Type I Diabetes Mellitus is More Sensitive to Ang II Due to Increased AT1 Receptor Expression in Afferent Arteriole
Authors: Zhang Jie , Lu Yan , Wang Shaohui , Zhang Gensheng , Wei Jin , Yip Kay-Pong , Liu Ruisheng
Published: March 2022

The prevalence of hypertension is about twice in diabetic subjects than that in non-diabetics. In the presence of hypertension, the progress of diabetic nephropathy has been shown to be more severe and rapid. But the mechanisms for the development of hypertension in diabetes have not been elucidated. We hypothesized that angiotensin II receptor type 1 (AT1 receptor) expression level in renal afferent arteriole is elevated, which enhances the responses to angiotensin II (Ang II) stimulation and contributes to the development of hypertension in diabetes.

First, we measured mean arterial pressure (MAP) with telemetry in diabetic and non-diabetic mice with Ang II infusion for 8 weeks. MAP was 26.7 mmHg higher in diabetic mice (157.4±11.3 mmHg) than in non-diabetic mice (130.7±6.4 mmHg). Next, we induced 2-kidney-1-clip Goldblatt hypertension in diabetic mice and non-diabetic mice and measured MAP with telemetry for 8 weeks. MAP was 17.6 mmHg higher in diabetic mice (151.1±9.5 mmHg) than in non-diabetic mice (133.5±4.2 mmHg), indicating blood pressure of diabetic mice is also more responsive to both exogenous and endogenous Ang II than no-diabetic mice.

To investigate the mechanisms of the enhanced sensitivity to Ang II, we dissected afferent arterioles from non-diabetic mice and diabetic mice and measured the expression of AT1 receptor mRNA by real-time PCR. AT1 receptor mRNA level was 6 times higher in diabetic mice that in non-diabetic control. AT1 receptor protein abundance in renal cortex was also 5 times higher in diabetic mice than controls.

In conclusion, the blood pressure of the diabetic mice are more sensitive to Ang II than non-diabetic controls. Increased AT1 receptor expression in the afferent arterioles may contribute to the development of hypertension in diabetes.
Abstract P103: Cellular Stretch Induces Higher Intracellular Calcium Increases in Dahl Salt-Sensitive Than Salt-Resistant Rat Thick Ascending Limbs: Role of Transient Receptor Potential Vanilloid 4
Authors: Cabral Pablo , Gonzalez-Vicente Agustin , Hong Nancy , Garvin Jeffrey
Published: March 2022

The thick ascending limb (TAL) reabsorbs 30 % of the filtered NaCl playing a pivotal role in the regulation of salt homeostasis. Abnormal reabsorption of NaCl in this segment causes salt-sensitive hypertension. We showed that flow-induced cellular stretch stimulates superoxide production in epithelial cells from TALs. Dahl salt-sensitive rat (DSS) TALs produce more superoxide than those from salt-resistant rats (DSR). Mechanical stimulation exerted by cellular stretch increases intracellular calcium (Cai) in several types of cells. We have shown that luminal flow-induced increases in Cai is mediated by Transient Receptor Potential Vanilloid (TRPV4). Therefore, we hypothesized that cellular stretch increases Cai in TALs from DSS rats more than those from DSR rats due to elevated TRPV4 activity. We measured Cai using the calcium-sensitive dye Fura-2 in isolated, perfused DSR and DSS TALs. Cellular stretch increased Cai by 243 ± 51 nM in DSS TALs (n=9; p<0.0008 vs. no stretch conditions) and by 124 ± 27 nM in DSR TALs (n=10; p<0.0005 vs no stretch conditions). Cellular stretch-induced Cai increases were significantly higher in the DSS group (p<0.05 vs DSR). When these animals were fed a high-salt diet (4% NaCl) similar responses were obtained. Cellular stretch increased Cai by 236 ± 58 nM in DSS TALs (n=7; p<0.004 vs no stretch conditions) and by 92 ± 15 nM in DSR TALs (n=6; p<0.0008 vs no stretch conditions). To study whether DSS and DSR TALs express different amounts of TRPV4 we performed Western blots. We found that TRPV4 protein expression was similar in TALs lysates from both strains fed either a control standard or a high-salt diet. We then tested the effects of transfecting DSS TALs with TRPV4-small hairpin (sh) RNA on stretch-induced Cai increases. Under these circumstances, the difference in stretch-induced Cai increases between DSS and DSR TALs was not significant (75 ± 15 nM in DSS vs 56 ± 28 nM vs DSR, n=4 for each group) . We conclude that DSS TALs are more sensitive to increases in cellular stretch than DSR TALs possibly due to enhanced TRPV4 activity.
Abstract P105: ACE2 Activator, Diminazene Aceturate, Reduces Adiposity, but Preserves Lean Mass in Young and Old Rats
Authors: Bruce Erin , Sakarya Yasemin , Kirichenko Nataliya , Toklu Hale Z , Morgan Drake , Tumer Nihal , Carter Christy S , Scarpace Philip J
Published: March 2022

Obesity is severely debilitating, and increases the risk for cardiovascular events. Recent evidence suggests increased Ang-(1-7)/Mas activity in obese animal models leads to significant reductions in body weight. We hypothesize that activation of ACE2, via Diminazene Aceturate (DIZE), will significantly reduce body weight of both young and aged rats fed 60% high fat diet.

Male Fisher 344 x Brown Norway rats, ages 4 (n=12) and 23 months (n=17) were fed 60% high fat diet for one week, whereupon, animals were further divided, and given either 15mg/kg/day DIZE s.c. (Young DIZE, n=6; Old DIZE, n=9) or vehicle (Young Control, n=6; Old Control, n=8). Body weight and food intake were measured throughout the experiment. Time Domain-Nuclear Magnetic Resonance (TD-NMR) was used to assess body composition 1 week and 3 weeks after the start of DIZE treatment.

DIZE treatment resulted in a significant reduction of food intake and change in body weight in both young and old animals. TD-NMR results determined the weight-loss was primarily a result of decreased body fat percentage, with a preservation of lean mass as indicated by a reduced fat/lean mass ratio. Tissue weights at the time of sacrifice confirm the significant loss of white adipose tissue (WAT), with no significant difference in tibialis anterior (TA) muscle weights. Importantly, when assessing heart weights, we observed a significant reduction in aged rats treated with DIZE when compared to age matched controls, suggesting a prevention of cardiac hypertrophy associated with obesity. Our data suggests DIZE may be a useful tool in the treatment of obesity along with associated co-morbidities.
Abstract P108: Caveolin-1 is Required for Vascular Remodeling Induced by Angiotensin Ii
Authors: Kawai Tatsuo , Obama Takashi , Takayanagi Takehiko , Forrester Steven J , Elliot Katherine J , Crawford Kevin , Eguchi Satoru , Rizzo Victor
Published: March 2022

We have recently reported that caveolin-1 (Cav1) enriched membrane microdomains in vascular smooth muscle cells (VSMC) mediates a metalloprotease ADAM17-dependent EGF receptor (EGFR) transactivation, which is linked to vascular remodeling induced by AngII. We have tested our hypothesis that Cav1, a major structural protein of caveolae, plays a critical role in AngII-mediated vascular remodeling via regulation of ADAM17 and EGFR. 8 week old male Cav1-/- and control Cav+/+ wild-type mice (WT) were infused with AngII (1 μg/kg/min) for 2 weeks to induce vascular remodeling and hypertension. Upon AngII infusion, histological assessments demonstrated medial hypertrophy and perivascular fibrosis of coronary and renal arteries in WT mice compared to sham-operated mice. The AngII-infused WT mice also showed a phenotype of cardiac hypertrophy with increased HW/BW ratio (mg/g: 8.0±0.6 vs 5.7±0.7 p<0.01) compared with sham-operated WT control. In contrast, vascular remodeling but not cardiac hypertrophy were attenuated in Cav1-/- mice with AngII infusion; HW/BW ratio (8.6±0.5 vs 6.4±0.2 p<0.05) compared to sham-operated mice. However, AngII induced similar levels of hypertension in both WT and Cav1-/- mice as assessed by telemetry (MAP mmHg: 142±9 vs 154±20). AngII infusion in WT mice enhanced ADAM17 and phospho-Tyr EGFR staining in heart and kidney vasculature, whereas AngII-infused Cav1-/- mice showed diminished ADAM17 and phospho-Tyr EGFR staining within the vasculature. In addition, IHC analyses revealed reduced vascular ER stress in heart and kidney samples of AngII-infused Cav1-/- mice compared to WT mice. Expression of Cav1 was predominantly observed within the endothelium and was enhanced upon AngII infusion in WT mice. These data suggest that Cav1, and presumably endothelial caveolae microdomains, plays a critical role in vascular remodeling via vascular induction of ADAM17 and activation of EGFR independent of blood pressure or cardiac hypertrophy regulation.
Abstract P111: Cross-Inhibition of the Vasorelaxing Effect of Ang-(1-9) and Ang-(1-7) in Aortic Rings
Authors: Resende Gabriel S , Ocaranza María P , Santos Robson A
Published: March 2022

Angiotensin-(1-9) is a nonapeptide formed by the hydrolysis of angiotensin I by ACE2 that seems to counter-regulate the classical RAS axis. The effect produced by Ang-(1-9), has an important role in the cardiovascular system promoting effects similar to those of Ang-(1-7). As vasodilation, anti-hypertrophic action in cardiomyocytes and antihypertensive effect. Because it has been reported that the Ang-(1-9) effects are independent of the Ang-(1-7) receptor Mas, in this study we investigated a possible synergistic additive effect of Ang-(1-9) and Ang-(1-7) in aortic rings.

Materials and methods:
The endothelium-dependent vasodilatory response to Ang-(1-7) and Ang-(1-9) (10-10 - 10-6 moles/l) alone or in combination was tested in aortic rings taken from Sprague-Dawley rats, pre-contracted with phenylephrine (0.1 umoles/L).

Results:
In aortic rings from SD rats, Ang-(1-7) and Ang-(1-9) induces vascular relaxation (Emax= 13.8±4.5% and 15±2.5%, n=5, respectively). Interestingly, pre-incubation with Ang-(1-9) abolished the vasodilatation induced by Ang- (1-7) (Emax= -2.7 ±1.4%, n=5). Likewise, pre-incubation with Ang-(1-7) abolished the vasorelaxation induced by Ang- (1-9). Indeed in aortic rings pre-incubated with Ang-(1-7),addition of Ang-(1-9) induced contraction instead of relaxation of the aortic rings (Emax= -10±1.7%, n=5). To test the effect of acute combination of the peptides in aortic rings, Ang-(1-7) was prepared in a 10-7M solution of Ang-(1-9) or vice-versa. As observed with pre-incubetion, the vasodilator effect induced by Ang (1-7) or Ang-(1-9) was abolished (Emax= -2.9±2.6 %, n=6 and 4±1.3%, n=6, respectively). These results suggest that these two peptides counter-regulate the vascular effects of each other. Whether these phenomenon in due to receptor internalization or to a signaling-related mechanism, remain to be clarified.
Abstract P121: Vascular Protective Effects of Telomerase Activity Are Independent of Nuclear Function
Authors: Ebben Johnathan D , Hockenberry Joseph , You Ming , Beyer Andreas M
Published: March 2022

Rationale:
The contributions of nuclear telomerase activity to cardiovascular protection we have previously described are not defined. We used a novel inhibitor of nuclear telomerase activity (nucTERT I) to test the hypothesis that nuclear telomerase activity is not critical to the protective effects of telomerase.

Methods and Results:
To confirm the effect of nucTERT I cultured cells were treated for 24 h and subjected to standard cell fractionation procedures. Telomerase activity (TRAP assay) confirmed decreased nuclear telomerase activity after treatment. nucTERT I reduced nuclear telomerase activity (44% +/- 7) compared to untreated control.

To evaluate the effect of nucTERT I on vascular function, human microvessels were dissected from discarded surgical tissue from patients with no history of coronary artery disease and used for videomicroscopy. Vessels were challenged with ANG II and their ability to dilate in response to flow and acetylcholine was evaluated. Vessels pre-treated with nucTERT I (24 h) were significantly less sensitive to ANG II- induced endothelial dysfunction when stimulated with either acetylcholine or increased flow. The telomerase activity inhibitor, BIBR1532, abrogated these effects (figure).

Conclusions:
Our data suggest that nuclear telomerase activity is not necessary for the protective effects of telomerase on ANG II induced endothelial dysfunction. Treatment with BIBR1532 confirms that vascular stress resistance is conferred by catalytically active telomerase. These data suggest that modulation of telomerase may be a useful strategy in the treatment of vascular disease.
Abstract P136: What is the Impact of Arterial Stiffness on Brain's Health
Authors: Girouard Helene , Obari Dima , Muhire Gervais , Vallerand Diane , Ferland Guylaine , Sadekova Nataliya
Published: March 2022

Arterial stiffness is an important risk factor for cognitive decline. However, its specific effects on brain homeostasis are unknown. Hence, the objective of the study is to explore the effects of arterial stiffness on brain's health, especially on oxidative stress, inflammation, cerebrovascular regulation and cognitive functions.

Approach and Results:
Arterial stiffness was induced by applying calcium chloride to carotid arteries of C57BL6 male mice. The control group received sodium chloride. Cerebral inflammation was assessed by quantifying immunoreactivity to activated glia markers ; Iba-1, CD68 and s100β. Oxidative stress was determined with dihydroethidium. Cerebral blood flow (CBF) was monitored by laser-Doppler flowmetry in anesthetized mice equipped with a cranial window and spatial memory was tested using the Morris water maze. Results show that arterial stiffness activates microglia in the hippocampus, and astrocytes in the hippocampus and the frontal cortex. Superoxide anion production was elevated in the hippocampus of these mice. Arterial stiffness attenuated the CBF increase produced by stimulation of the vibrissae or by topical application of the endothelium-dependent vasodilator acetylcholine. The results indicate learning and spatial memory deficit induced by arterial stiffness in comparison to the sodium chloride group.

Conclusions:
This study shows that arterial stiffness, induced by carotid calcification, leads to cerebral inflammation and increased oxidative stress mainly in the hippocampus. Arterial stiffness also alters CBF regulation and cognitive functions. This suggests that arterial stiffness has an impact on cerebral homeostasis and should be considered as a therapeutical target for the prevention of cerebral dysfunctions in the aging population.
Abstract P143: The Angiotensin AT2 Receptor Agonist Compound 21 Is a Low Affinity Thromboxane A2 Receptor Antagonist
Authors: Steckelings Ulrike M , Fredgart Maise , Leurgans Thomas , Stenelo Martin , Nybo Mads , Rasmussen Lars M , De Mey Jo G
Published: March 2022

Objective:
The purpose of this study was to investigate potential vasorelaxant effects of angiotensin AT2-receptor stimulation by the specific AT2-receptor agonist C21 in pericardial resistance arteries from cardiovascular disease patients.

Methods:
Parietal pericardium was obtained during coronary artery bypass-grafting and/or valve replacement in patients. Pericardial arteries from pigs and mesenteric arteries from C57BL-6J and AT2R deficient (AT2R-/y) mice were used for comparison.

Isolated arteries were mounted onto wire myographs, pre-contracted with either K+, phenylephrine, endothelin-1 (ET-1) or the thromboxane agonist U46619, and the relaxing effect of C21 (0.1nM - 10μM) was investigated in the absence or presence of 3nM valsartan (AT1R antagonist), 10μM PD123319 or 10μM M132 (AT2R antagonists).

Results:
C21 significantly relaxed ET-1 contracted porcine arteries (Emax -37 ± 6 %; -0.87 vs -0.45 N/m relaxation; N=6; P<0.05, two-way ANOVA) and phenylephrine contracted mouse mesenteric arteries (Emax -52 ± 10 %; -0.26 vs -0.09 N/m relaxation; N=6; P<0.01, two-way ANOVA), and this was inhibited by PD123319 or M132 and absent in arteries from AT2R-/y. C21 significantly induced vasorelaxation in U46619 contracted vessels from all species and vascular beds (e.g. in human pericardial arteries: Emax -78 ± 10 %; -0.67 vs -0.07 N/m relaxation; N=17; P<0.01, two-way ANOVA), and this effect was not blocked by AT2R antagonists and still present in AT2R-/y. C21 further inhibited U46619 induced platelet aggregation. An Arrestin Biosensor Assay revealed that C21 binds to the TXA2 receptor with a Ki of 3.74 μM.

Conclusion:
Depended on species, vascular bed and contractile stimulus, C21 relaxes resistance-sized arteries by AT2R stimulation or by TXA2 antagonism. These data together with the low affinity binding of C21 to the TXA2 receptor and its effect on platelet aggregation strongly suggest that C21 is not only a high affinity, selective AT2R agonist, but also a low affinity TXA2 receptor antagonist.
Abstract P152: Acute Effect of Unilateral Unipolar Electrical Carotid Sinus Stimulation in Patients with Treatment-Resistant Arterial Hypertension
Authors: Heusser Karsten , Brinkmann Julia , Menne Jan , Kaufeld Jessica , Linnenweber-Held Silvia , Beige Joachim , Wilhelmi Mathias , Diedrich André , Haller Hermann , Jordan Jens , Tank Jens
Published: March 2022

Electrical carotid sinus stimulation has been developed for treatment of resistant arterial hypertension. The first-generation device (Rheos™) relying on bilateral placement of bipolar electrodes acutely reduced muscle sympathetic nerve activity (MSNA) and blood pressure (BP) but is no longer available. The second-generation device (Neo™) utilizes a smaller unilateral unipolar disk electrode to reduce invasiveness while saving battery life. We tested acute effects of the latter on BP and MSNA.

We studied 18 treatment-resistant hypertensive patients (9 women, 53±11 years, 34±5 kg/m
2
) on stable medication who had been implanted with the second-generation device. We assessed acute BP (oscillometry), heart rate (HR, ECG), and MSNA (microneurography) responses to electrical stimulation in the supine position.

Without stimulation, BP was 165±31/91±18 mmHg, HR was 75±17 bpm, and MSNA was 48±14 bursts/min. Acute stimulation with intensities producing side effects that were tolerable in the short term elicited variable changes in systolic BP (SBP: -16.9±15.0 mmHg, range: 0.0 to -40.8 mmHg, p=0.002), HR (-3.6±3.6 bpm, p=0.004), and MSNA (-1.9±5.3 bursts/min, p=0.194). Stimulation intensities had to be lowered in 12 patients to avoid side effects at the expense of efficacy (SBP: -6.3±7.0 mmHg, range: 2.8 to -14.5 mmHg, p=0.028; HR: -1.5±2.3 bpm, p=0.078; comparison against responses with side effects). Reductions in diastolic BP and MSNA (total activity) tended to be correlated (r
2
=0.202, p=0.093).

In our patient cohort, unilateral unipolar electrical baroreflex stimulation acutely lowered BP. Side effects may limit efficacy. The novel approach should be tested in a controlled comparative study.
Abstract P153: Impact of Antibiotic Treatment on Antihypertensive Drug Adherence
Authors: Kassi Maria , McCallum Linsay , Muir Scott , Touyz Rhian , Dominiczak Anna F , Padmanabhan Sandosh
Published: March 2022

Background:
Suboptimal adherence reduces the effectiveness of antihypertensive therapy and is recognised as a significant obstacle in achieving better patient outcomes. The impact of concomitant therapy on adherence levels is unknown. The aim of this study was to identify changes in antihypertensive drug adherence after antibiotic treatment.

Methods:
We analysed refill prescriptions (2004-2013) from 361,021 patients who attended 2 hospitals in Glasgow. Antihypertensive drug adherence over the preceding and following 12 months of first antibiotic therapy in each patient were analysed. Drug adherence was measured as drug usage over the study period and calculated from the amount of drug dispensed and the defined daily dose for each antihypertensive drug. Comparisons were made using paired-t tests.

Results:
There were 190,699 subjects prescribed amoxicillin, 63,292 ciprofloxacin, 69,756 clarithromycin and 94,832 trimethoprim. The mean difference in adherence for each antihypertensive drug before and after amoxicillin, clarithromycin, ciprofloxacin and trimethoprim are presented in the Figure. There is a general reduction in adherence following any antibiotic therapy for the antihypertensive drugs studied, with bendrofluomethiazide and atenolol showing the least changes in adherence and doxazosin and ramipril associated with the highest decreases in adherence. No differences between antibiotics was observed for any antihypertensive drug.

Conclusion:
A single course of antibiotic therapy can have a sustained adverse impact on antihypertensive drug adherence over the following year. The impact of this on BP control and outcomes need to be further study.
Abstract P157: Tractography of White Matter Connections Predicts for Vascular Cognitive Impairment in Hypertensive Patients
Authors: Carnevale Lorenzo , Selvetella Giulio , Cugino Daniela , Grillea Giovanni , Lembo Giuseppe , Carnevale Daniela
Published: March 2022

Vascular cognitive impairment (VCI) results by several vascular risk factors and, particularly, hypertension (HTN). The identification of early changes associated with later development of dementia is demanding. Great part of research has primarily focused on brain changes occuring in grey matter. However, more recent data highlighted that HTN may determine cognitive decline, even before manifest neurodegeneration. Diffusion tensor imaging (DTI) on magnetic resonance, opened the possibility to predict white matter connections that correlate with specific cognitive functions. In this study, we used DTI and cognitive assessment (CA), in order to identify a regional pattern of fractional anisotropy (FA) changes that could predict for VCI in hypertensive patients (HT).

We have examined 15 HT (moderate to severe, with antihypertensive medications) vs 15 normotensive (NT), subjecting them to DTI and CA. HT had significant higher SBP (138±4 vs 118±3 in NT) and DBP (87±2 vs 75±2 in NT) (p<0.001), displayed a significant LV hypertrophic remodeling (LVM/BSA 112±5 vs 83±3 for NT) (p<0.0001), with a significant moderate increase in albuminuria (15.7±2.6 mg/24h vs 8.8±1.6 for NT) (p<0.03). When subjected to CA, HT had significantly worsen performance on both MoCA (22.66±0.97 vs 26.21±0.57 NT) and Stroop Test (34.50±3.87 vs 17.75±2.57 NT) (p<0.01). Conversely, tests regarding Verbal Fluency and Instrumental Activities of Daily Living revealed normal performance of HT, thus indicating a selective impairment of memory. Brain imaging showed that, while none of the patients had abnormal signal intensity on T1/T2-weighted MRI,

DTI indices FA were significantly reduced in HT as vs NT. In particular, HT had lower FA in projection fibers related to impairment for non-verbal materials (Anterior Thalamic Radiation: 0.358±0.012 vs 0.330±0.006, p<0.05), association fibers involved in executive functioning and emotional regulation (Superior Longitudinal Fasciculus: 0.388±0.013 vs 0.356±0.007, p<0.05), limbic system fibers involved in attention tasks (cingulate gyrus: 0.364±0.009 vs 0.328±0.010, p<0.01).

Our data highlight a novel paradigm of combined DTI/CA of HT patients, capable to identify, with great sensitivity, predictive signs of HTN-induced VCI.
Abstract P161: Angiotensin Receptor Blockade Decreases Glutamate-induced Brain Inflammation
Authors: Saavedra Juan , Elkahloun Abdel G , Hafko Roman
Published: March 2022

Chronic Kidney Disease (CKD) is very frequently associated with brain inflammation and cell injury leading to cognitive loss. At present a combined treatment of related kidney and brain injury has never been proposed. We focused on glutamate-induced excitotoxicity and neuronal inflammation, and on the effects of candesartan, a renoprotective Angiotensin Receptor Blocker (ARB) ameliorating hypertension and diabetes-induced kidney damage, and with accompanying neuroprotective efficacy. Primary cerebellar granule cells (CGC) were exposed to 100 μM glutamate and pre-treated for one hour with candesartan at neuroprotective concentrations (10 μM). Gene expression was quantified by qPCR. Candesartan significantly reduced glutamate-induced inflammation. Multiple group comparisons were performed by one-way ANOVA followed by Newman-Keuls post-test. Exposure to glutamate significantly reduced neuronal viability while up-regulating the expression of multiple genes on pro-inflammatory pathways, including Toll-like receptor 7 (Tlr7) and Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2, p49/p100 (Nfkpb2). * p<0.01, ** p< 0.05, vs control; # p< 0.01 vs glutamate. In all cases, pretreatment with candesartan completely prevented glutamate-induced neurotoxicity. Our results indicate that Angiotensin II receptor blockade with candesartan is strongly and directly neuroprotective, significantly ameliorating neuronal injury as a result of glutamate excitotoxicity. These results support the use of candesartan and other ARBs for the concomitant treatment of CKD and associated neuronal injury.
Abstract P162: Nicotine via Cd36, Cox2 and Oxidative Stress Promotes Podocyte Injury: Link Between E-cigarettes and Renal Disease Progression?
Authors: Raij Leopoldo , Tian Ruxia , Zhou Ming-Sheng
Published: March 2022

Cigarette smoking (CS) accounts for 175,000 annual CV deaths in US. CKD is a major CV risk factor. Epidemiologically link between CS proteinuria and progression of diabetic and of hypertensive nephropathy is documented. We showed that Nicotine (NIC) in concentrations achieved in CS and E- smokers a)Increased proteinuria (100%) renal Nox4 & Nitrotyrosine in diabetic db/db mice (AJP ‘10) and b) Promoted in human macrophages O
2
-
production and foam cell formation associated with upregulation of B scavenger receptor CD36 and oxLDL uptake (AJP’13). Podocytes (POD) are vulnerable to diabetes and to hypertension; POD injury results in detachment and glomerulosclerosis. We demonstrated in human POD NIC receptors α2, 3,4 and β3
Methods:
We treated human POD with NIC, 100nmol/L, a concentration attained in serum of CS and E-Cigarettes smokers, and determined O
2
-
production with lucigening; some POD were pre-incubated with Hexametonium: blocker of NIC receptors; DPI: inhibitor of NADPH; Catalase: inhibitor of H2O2 and AICAR: activator of AMPK, a suppressor of NADPH. By Western Blot (WB) we determined in Control and NIC exposed POD, CD36, COX2 known to induce POD injury (JASN ‘08) and Synaptopodin (Synpo) a stabilizer of POD actin skeleton. In POD incubated with 50ug/ml oxLDL we measured POD apoptosis (APOPT) by flow cytometry. Stat. ANOVA- Bonferonni’s- Scheffe’s.
Results:
NIC increased POD O
2
-
production 225% (200 to 450 CPM/ug); Synpo expression was reduced 50% (1to 0.5) (p< 0.05). DPI and AICAR prevented Synpo reduction and increase in O
2
-
(p<0.05). NIC upregulated expression of COX2 300% (0.5-1.5) and CD36 40 % (1-1.4) (p<0.05). DPI, Catalase, and Hexametonium inhibited CD36 and COX2 upregulation. The COX inhibitor NS-398 prevented O
2
-
production and CD36 upregulation .POD incubation with oxLDL plus NIC significantly increased POD oxLDL uptake and APOPT 10% over baseline (2%),(p<0.05) both inhibited by CD36-SiRNA.
Conclusion:
NIC
activated
in human POD a reciprocal
interplay
between NADPH oxidases and COX2 that increased O2- production, reduced Synpo and upregulated the B scavenger receptor CD36 that, via increased uptake of oxidized LDL, promoted podocyte
apoptosis.
We surmise that these novel findings implicate NIC as a risk factor for CVD and CKD progression.
Abstract P178: Depletion of Gamma-Delta T Cells Protects Against Toll-Like Receptor-Induced Preeclampsia in Mice
Authors: Bounds Kelsey R , Chiasson Valorie L , Tobin Richard P , Newell-Rogers M K , Mitchell Brett M
Published: March 2022

Preeclampsia (PE), a hypertensive disorder of pregnancy, is associated with vascular endothelial dysfunction and excessive immunity and inflammation. However, it is unclear which innate immune cells propagate the pro-inflammatory state. Gamma-delta T (gdT) cells can secrete tolerogenic anti-inflammatory cytokines or cytotoxic pro-inflammatory cytokines depending on their activation status. gdT cells from women with PE produce significantly more IFNg and perforin and are less susceptible to apoptosis than gdT cells from normal pregnant women. We hypothesized that Toll-like receptor (TLR) activation in gdT cells induces inflammation and causes PE-like features in mice and that gdT cell KO mice would be resistant to developing TLR-induced PE-like features. Activation of splenocytes isolated from day 14 normal pregnant mice with the TLR3 agonist poly I:C or the TLR7 agonist R837 for 24 hours significantly increased gdT cells as well as IFNg and TNFa production. We have reported that poly I:C or R837 treatment of normal pregnant mice elicits a pregnancy-dependent PE-like syndrome by inducing a pro-inflammatory immune response. Pregnant poly I:C-treated and R837-treated mice had significantly increased splenic levels of gdT cells and plasma levels of IFNg and TNFa compared to pregnant vehicle-treated mice. Pregnant gdT cell KO mice treated with poly I:C or R837 did not develop hypertension or endothelial dysfunction. These data demonstrate that gdT cells mediate the TLR-induced PE-like features in mice and depletion of gdT cells may reduce the severity of PE in women.
Abstract P180: Dopamine D2 Receptors Regulate Wnt Signaling and Apoptosis in Human Renal Proximal Tubule Cells
Authors: Han Fei , Zhang Yanrong , Konkalmatt Prasad , Jones John E , Gildea John , Felder Robin A , Jose Pedro A , Armando Ines
Published: March 2022

Previous work from our laboratory indicates that the dopamine D2 receptor (D2R) in the kidney has a direct role in regulating renal inflammation and injury and blood pressure. Some common single nucleotide polymorphisms (D2R SNPs; rs 6276, 6277, and 1800497) in the human
DRD2
gene are associated with decreased D2R expression and function. Immortalized renal proximal tubule cells (RPTCs) from subjects carrying D2R SNPs (RPTC-D2R SNPs) express less D2Rs than RPTCs carrying no D2R SNPs (RPTC-D2R WT) (62±4 vs 100±6%; P<0.04) and a pro-inflammatory and pro-fibrotic phenotype with markers of epithelial mesenchymal transition. RPTC-D2R SNPs showed increased apoptosis compared with RPTC-D2R WT (11± 0.8 vs 2.3±0.4% TUNEL positive cells, P<0.01, n=5/group). We hypothesized that the D2R regulates renal cell survival through effects on Wnt signaling. We found that Wnt3 expression was increased in RPTC-D2R SNPs compared with RPTC-D2R WT (mRNA: 2.6±0.35 vs 1±0.11 fold; P<0.05; protein: 133±4 vs 100±5%; P<0.05). RPTC-D2R SNPs showed activated Wnt3/β-catenin signaling pathway demonstrated by decreased β-catenin phosphorylation (64±4 vs 100±8%; P<0.05) and increased expression of downstream pro-apoptotic factors Bax (136±4.6 vs 100±5%, P<0.05) and FasL (128±5.6 vs 100±6.5%, P<0.05). Silencing D2R in RPTC-D2R WT (siRNA; 0.30±0.02 vs 1±0.07 fold, P<0.05) increased Wnt 3 expression, decreased β-catenin phosphorylation, and increased expression of Bax and FasL. By contrast, treatment of RPTC-D2R WT with a D2R agonist (quinpirole,1μM, 24h) or transfection of RPTC-D2R SNPs with a
DRD2
which restored D2R expression decreased Wnt3 expression, increased β-catenin phosphorylation, and decreased Bax and FasL expression. Moreover, Wnt3 silencing (siRNA) in RPTC-D2R SNPs increased β-catenin phosphorylation (132±5 vs100±9%, P<0.05), decreased Bax and FasL expression, and reduced the number of apoptotic cells (6±1.0 vs12± 0.9 % TUNEL positive cells, P<0.01). Our results indicate that D
2
R function is important in the regulation of the Wnt pathway and that the alterations in D
2
R function result in modifications in the pathway potentially leading to fibrosis, cell death, and hypertension. These results may have clinical relevance for subjects bearing D2R SNPs.
Abstract P188: Mitochondrial Redox Phenotype Contributes to Angiotensin Induced Hypertension
Authors: Neville Amber , Leskov Igor , Krzywanski David
Published: March 2022

Oxidant stress contributes to the initiation and progression of hypertension by contributing to endothelial dysfunction and/or causing perturbations in nitric oxide homeostasis. Differences in mitochondrial function may contribute to this process and provide insight into why age of onset and clinical outcomes differ amongst individuals form distinct ethnic groups. We have previously demonstrated that variations in normal mitochondrial function and oxidant production exist in endothelial cells from individuals of Caucasian and African American ethnicity and that this variation contributes to endothelial dysfunction. To model these distinct mitochondrial redox phenotypes we used the C57Bl/6J (6J) and C57Bl/6NJ (NJ) that also display unique mitochondrial redox phenotypes due to the differential expression nicotinamide nucleotide transhydrogenase. Interestingly, we found that the 6J animals had a significantly higher systolic blood pressure compared to the NJ animals (116 ± 0.96 vs 102 ± 2.39 mmHg) that was exacerbated by angiotensin II induced hypertension (159 ± 8.25 vs 136 ± 4.33 mmHg). Vascular oxidative stress was assessed in these animals by evaluating mitochondrial oxygen utilization and oxidant production in primary aortic endothelial cells (MAEC) from these animals. Bioenergetic analysis indicates that compared to NJ, 6J MAEC utilized significantly less oxygen for ATP production, possess a lower maximal respiratory capacity (17.31 ± 1.84 vs 41.87 ± 2.95 pmol O
2
/min/ug protein), and have reduced electron leak. 6J MAEC also produce more superoxide compared to NJ in response to angiotensin II stimulation (0.08 ± 0.004 vs 0.11 ± 0.006 nmol/mg protein). Additionally, isolated renal mitochondria and vessel myography indicated differences in respiratory coupling ratios and endothelial dependent vasodilation amongst the groups. Taken together, these data indicate that differences in “normal” mitochondrial function amongst ethnic groups could contribute to a unique mitochondrial redox phenotype that influences individual susceptibility by contributing to endothelial dysfunction, providing important insights into the mechanisms that contribute to the development of hypertension.
Abstract P193: Upregulation of Brain Renin-angiotensin System and Inflammation Mediates Leptin Sensitization of Angiotensin Ii-induced Hypertension
Authors: XUE BAOJIAN , Yu Yang , Guo Fang , Beltz Terry G , Felder Robert B , Johnson Alan K
Published: March 2022

Leptin, an adipocyte-derived hormone, contributes to the increase in blood pressure (BP) associated with obesity. Obesity has also been shown to promote the renin-angiotensin system (RAS) activity and inflammation in the hypothalamus that increase BP and sympathetic activity. Our previous studies using an Induction-Delay-Expression experimental design demonstrated that a central leptin pretreatment resulted in an enhanced hypertensive response to subsequent treatment with angiotensin (ANG) II. The present study tested whether this central leptin sensitization of ANG II-induced hypertension is mediated by an increase in RAS acitivity and inflammation in the brain. Male rats prepared for telemetry BP recording and were pretreated during Induction with central leptin (20 ng/kg/min, ICV) alone or with either ANG II type 1 receptor (AT1R) antagonist irbesartan (125 μg/d) or the TNF-α synthesis inhibitor pentoxifylline (10 μg/h) for one week. After one week Delay, rats were treated during Expression with subcutaneous ANG II (120 ng/kg/min) for 2 weeks. The animals pretreated with central leptin responded with enhanced hypertension to ANG II ([[Unable to Display Character: ∆]]39.3±3.8 mmHg vs. [[Unable to Display Character: ∆]]21.2±5.2 mmHg). Central infusion of pentoxifylline or irbesartan during Induction blocked the sensitization produced by central leptin. RT-PCR analysis of tissue from the lamina terminalis indicated that leptin up-regulated mRNA expression of several components of the RAS and proinflammatory cytokines including AT1R, angiotensin converting enzyme, TNF-α and interleukin 1β. The results indicate that leptin-induced sensitization of ANG II-elicited hypertension is mediated by upregulation of central RAS and proinflammatory cytokines.
Abstract P208: Effect of Rosiglitazone on Renal Neprilysin Activity and Protein Expression in Db/db Diabetic Mice
Authors: Alawi Laale , Emberesh Sana , Elased Khalid
Published: March 2022

Alteration in renin-angiotensin system has been implicated in the pathophysiology of diabetic kidney disease. The deleterious actions of angiotensin II (Ang II) could be antagonized by the formation of Ang (1-7) partly generated by the actions of angiotensin converting enzyme 2 (ACE2) and neprilysin (NEP). NEP is a member of the zinc-containing metallopeptidase family, and has a main role in the degradation of several peptides, including natriuretic peptides, bradykinin, amyloid beta, and Ang I. Our previous studies demonstrated increased shedding of renal ACE2 in db/db mouse model of type 2 diabetes. We also showed that treatment with the PPAR_ agonist, rosiglitazone normalized hyperglycemia, decreased urinary ACE2 and attenuated albuminuria in db/db mice. The aim of the study was to test the hypothesis that hyperglycemia down-regulates renal NEP in db/db diabetic mice and that treatment with rosiglitazone will normalize renal NEP expression and activity. Seven-week-old db/db male mice were subjected to rosiglitazone treatment (20 mg/kg/day) for 10 weeks. Treatment with rosiglitazone significantly lowered blood glucose levels in db/db mice (p<0.001). Western blot analysis and immunohistochemistry demonstrated a significant decrease in renal and urinary NEP protein expression in 17wk db/db mice compared to lean control mice (p<0.0001). Treatment of db/db mice with rosiglitazone attenuated albuminuria, increased renal NEP protein expression and activity. In conclusion, the renoprotective effects of rosiglitazone could be mediated by up-regulation of renal NEP expression and activity in db/db mice. Alteration in the balance between Ang II and Ang (1-7) forming enzymes could contribute to the development of albuminuria in db/db diabetic mice.
Abstract P215: Angiotensin II-Mediated Cardiac Remodeling in cChAT Mice
Authors: Salgado Helio C , Tezini Geisa C , Roy Ashbeel , Prado Vania F , Gros Robert , Prado Marco A
Published: March 2022

It has been demonstrated that the cardiac non-neuronal cholinergic system (NNCS) plays a role in regulating cardiac homeostasis under physiological conditions. To examine a possible role played by NNCS upon cardiac remodeling, we submitted mice with genetic deletion of cardiomyocyte-specific choline acetyltransferase (cChAT) to chronic subcutaneous infusion of angiotensin II (Ang II). Male mice aged six months were assigned into four experimental groups: wild-type (WT) + Saline; WT + Ang II; cChAT + Saline and cChAT + Ang II. Ang II did not change the heart weight in WT mice (10.4±0.3 vs. 8.5±0.2 mg/mm in WT+Saline) but determined an increase in cChAT mice (12.8±1 vs. 8.8±0.6 mg/mm in cChAT+Saline). In WT mice, Ang II decreased left ventricular (LV) fractional shortening (23±2 vs. 35±2 % in WT+Saline) and LV ejection fraction (54±4 vs. 73±2 % in WT+Saline). However, cChAT+Ang II mice exhibited greater decrease in both LV fractional shortening (12±1.8 vs. 23±2 % in WT+Ang II) and ejection fraction (31±4 vs. 54±4 % in WT+Ang II). cChAT mice displayed cardiomyocyte hypertrophy (2002±116 vs. 1377±48 μm2 in WT+Saline) even when receiving saline. Ang II increased the cardiomyocyte surface area in both WT and cChAT mice; however, the cChAT mice exhibited greater myocyte hypertrophy (3035±181 vs. 2603±122 μm2 in WT+Ang II). Hematoxylin and eosin staining revealed that Ang II promoted greater disruption of myocardial structure in cChAT mice. Additionally, Trichrome C staining revealed that WT+Ang II mice presented increased collagen deposition (2.86±0.2 vs. 0.45±0.07 % in WT+Saline); nevertheless, the fibrotic response in cChAT+Ang II mice was greater than that observed in WT animals (6.16±1.1 vs. 2.86±0.2 % in WT+Ang II). Therefore, mice with deficiency for ChAT displayed exacerbated ventricular dysfunction induced by chronic Ang II administration, providing support for a role of the NNCS in the progression of cardiac remodeling.
Abstract P218: Role of Metabolic Biomarkers in Early Detection of Diabetic Cardiomyopathy in West Virginian Population
Authors: Getty Morghan , Thompson Ellen , Jones Christopher I , Okhumale Paul I , Sodhi Komal
Published: March 2022

Background:
Diabetes Mellitus is a significant risk factor for heart failure. With the increasing global prevalence of diabetes, diabetic cardiomyopathy (DCM) represents a significant public health issue. Elevated levels of Insulin-like growth factor-binding protein 7 (IGFBP7) is an early-stage biomarker in DCM but studies are lacking on the association between IGFBP7 and DCM. Most patients with DCM remain asymptomatic until late-stage disease, therefore it becomes of utmost importance to identify biomarkers to detect early stage of DCM. The aim of this study is to determine if IGFBP7 and other biomarkers can detect the onset of preclinical DCM relative to echocardiographic irregularities exhibiting diastolic dysfunction.

Methods:
Eighty patients were grouped equally into four categories based on predetermined clinical diabetic and cardiac parameters:
Normal, Diabetes (DM), Diastolic Heart Failure (DHF)
, and
DM+DHF,
the last group being the preclinical DCM group.

Results:
Echocardiography images indicated severe diastolic dysfunction in patients with DHF + DM as compared to DHF patients alone. In the
DM
and
DM+DHF
groups, IL-6, TNF-alpha, isoprostane, and leptin were elevated compared to the control, as were clinical markers HDL, glucose and hemoglobin A1C. More importantly fibrotic markers IGFBP7 and TGF-β followed the same trend
.
The
Normal
group showed higher levels of beneficial biomarkers adiponectin and bilirubin, which were reduced in the
DM
and
DM+DHF
groups.

Conclusion:
This novel study demonstrates that West Virginia patients with early onset DCM were more likely to have elevated levels of detrimental biomarkers, including IGFBP7 and TGF- β, indicating potential diastolic dysfunction. The present study provides new and exciting evidence supporting the potential clinical applications of using IGFBP7 and other biomarkers in diagnosing early stage DCM in the West Virginia population.