bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P241: Involvement of Mitochondrial Telomerase Reverse Transcriptase in the Restoration of Nitric Oxide Levels by Angiotensin-(1-7) in the Dysfunctional Diabetic CD34
+
Cells
Authors: Joshi Shrinidh , Gomez Sam , Cantu Wendy , Quiroz Julio , Beyer Andreas , Garcia Charles A , Jarajapu Yagna P
Published: December 2018

Telomerase reverse transcriptase (TERT) has been shown to translocate to mitochondria (mtTERT), and decrease the generation of mitochondrial reactive oxygen species (mtROS). Angiotensin (Ang)-(1-7) stimulates vasoreparative functions in diabetic CD34
+
stem/progenitor cells in part by decreasing ROS levels and increasing nitric oxide (NO) bioavailability. This study tested the involvement of mtTERT in mediating the effect of Ang-(1-7) on NO levels in diabetic CD34
+
cells. CD34
+
cells were isolated from the peripheral blood mononuclear cells (MNCs) of nondiabetic subjects (ND), and type 1 or type 2 diabetic (DB) patients (male or female, age 48-76 years) (DB: HbA1C 6.5-11.2). Cells were treated with Ang-(1-7) (100nM) or Stromal derived factor-1α (SDF) and evaluated for NO, and mtROS levels by flow cytometry by using DAF-FAM and mitoSOX, respectively. Number of cells used was ≥5x10
4
per treatment. The Mean Fluorescence Intensity is expressed in arbitrary fluorescence units (AFIx10
5
). Inhibitors of Mas receptor and TERT, A779 and BIBR-1532, respectively, and decoy peptides that inhibit mitochondrial translocation or nuclear transport of TERT, mtXTERT and nucXTERT, respectively, were used. SDF (100nM) failed to stimulate NO generation in DB-cells (1.2±0.07 vs basal 0.8±0.05, n=10) compared to ND-cells (3.5±0.2 vs basal 1.6±0.2) (n=10, P<0.0001). Ang-(1-7) induced NO levels in ND- (2.9±0.2) and restored NO generation in DB-cells (2.2±0.1) (n=10). DB-cells have increased mitoROS (3.6±0.4) compared to ND-cells (1.7±0.1, P<0.001, n=7) and Ang-(1-7) normalized the mitoROS in DB-cells (1.8±0.3, n=7). The effects of Ang-(1-7) on NO and mitoROS levels were reversed by A-779 or BIBR-1532 (n=5). Decoy peptide mtXTERT not nucXTERT increased mitoROS in ND- (4.4±0.7 vs basal 1.8±0.2, P<0.01) and in DB-cells (12±1 vs basal 6±0.4, P<0.001) (n=5). The decreasing effect of Ang-(1-7) on mitoROS (1.6±0.3) was reversed by mtXTERT (10±0.9, P<0.001, n=5) but not by nucXTERT. Along similar lines, restoration of NO levels by Ang-(1-7) in DB-cells was reversed by mtXTERT but not by nucXTERT. These observations provide compelling evidence for the involvement of TERT in stimulating the vasoreparative functions of diabetic stem/progenitor cells by Ang-(1-7)/Mas pathway.
Abstract P250: Unveiling Binding Pocket Structure Of Mas Receptor And Its Interaction With Angiotensin-(1-7)
Authors: Bragina Maiia E , Cirauqui Nuria , Dal Peraro Matteo , Santos Robson A , Fraga-Silva Rodrigo A , Stergiopulos Nikos
Published: December 2018

Substantial amount of preclinical data points out Mas receptor as a promising pharmacological target to treat cardiovascular diseases. Here, we aimed to characterize Mas binding pocket and its interaction with Angiotensin-(Ang)-1-7, in order to provide insights for potential structure-based drug discovery. Homology modeling based on AT1 and AT2 crystal structures, combined with peptide docking (Schrodinger
TM
) and molecular dynamics simulations (Gromacs) were employed to generate hypothesis about crucial interactions in protein-ligand complex. Proposed model was further validated
in vitro
using HEK cells transiently transfected with wild-type Mas or mutant Mas (Arg245Ala). Mas-receptor activation was assessed by FOXO1 dephosphorylation at Ser256 after 5 minutes incubation with Ang-1-7 (10
-7
M). Model of Mas/Ang-(1-7) complex revealed electrostatic interaction between Ang-(1-7) C-terminus and Mas Arg245. Interestingly, while Ang-1-7 incubation reduced FOXO1 phosphorylation by 35 % in HEK-Mas wild type (1.000 vs. 0.654 A.U., p<0.01, n=8), this effect was abolished in HEK-Mas mutant (1.000 vs. 0.990 A.U.,
n.s.
, n=5). In sum, Arg245 of Mas is essential for the receptor activation, which is in accordance with A-779 antagonism (C-terminus modification of Ang-(1-7)), endorsing our proposed structure model. Further elucidation of the ligand-receptor interactions may drive rational drug discovery and lead to development of improved strategies for Mas targeting.
Abstract P255: Differential Regulation of Blood Pressure and Renal Injury by HDAC Inhibitor Mocetinostat in
Npr1
Gene-targeted Male and Female Mutant Mice
Authors: Kumar Prerna , Bloodworth Meaghan , Nguyen Christian , Gogulamudi Venkateswara R , Pandey Kailash N
Published: December 2018

The objective of the present study was to elucidate the systolic blood pressure (SBP) lowering effect of class I histone deacetylase (HDAC) inhibitor, mocetinostat (MGCD0103) by enhanced expression of natriuretic peptide receptor-A (NPRA) in gene (
Npr1
). Adult male and female
Npr1
heterozygous (1-copy;
Npr1
+/-
), wild-type (2-copy;
Npr1
+/+
), and gene-duplicated (3-copy;
Npr1
++/+
) mice were intraperitonealy injected with MGCD (2 mg/kg) for 2-weeks. The renal NPRA protein levels were significantly increased in the treated male (1-copy, 2.1-fold; 2-copy, 2.3-fold; and 3-copy, 1.5-fold; p < 0.05) and female (1-copy, 3.6-fold; 2-copy, 2.1-fold; and 3-copy, 1.7-fold; p < 0.05) mice compared with untreated animals. HDAC activity was significantly higher in male (1-copy, 24.4 ± 2.8; 2-copy, 15.9 ± 1.2; and 3-copy, 6.4 ± 0.7; ng/min/mg protein) than female (1-copy, 13.7 ± 1.4; 2-copy, 8.5 ± 0.9; and 3-copy, 4.7 ± 0.9, p < 0.05) mice; however, treatment with MGCD significantly attenuated HDAC activity in both male (~40%, p < 0.05) and female (~50%, p < 0.05) animals. Male mice exhibited significantly higher SBP than female mice and treatment with MGCD decreased SBP in both 1-copy male (106 ± 0.6 vs. control, 129 ± 1.9; mm Hg) and female (97 ± 2.2 vs. control, 110 ± 2.1, p < 0.001) mice. Significantly high urinary protein to creatinine ratio was detected in male compared with female mice which was attenuated by MGCD treatment in male mice (1-copy, 3.2 ± 0.4 vs. untreated 5.5 ± 04; 2-copy 2.5 ± 0.4 vs. untreated 4.6 ± 0.2; and 3-copy, 1.6 ± 0.3 vs. untreated 3.1 ± 0.4; p < 0.05). Lower creatinine clearance (μl/min) was observed in both male (1-copy, 51 ± 11 vs. 2-copy, 130 ± 14.0; p < 0.05) and female (1-copy, 55 ± 2.7 vs. 2-copy, 129 ± 6.4; p < 0.01) mice. However, treatment with MGCD increased creatinine clearance in male (105 ± 13; p < 0.05) and female (97 ± 8.4; p < 0.05) vs. control mice. The present results demonstrate that class I HDAC inhibitor, MGCD upregulates NPRA expression
in vivo
, lowers SBP, and repairs renal injury differentially in male vs. female mice. These findings will have important implications for treatment of hypertension and renal injury in humans in a sex-related manner.
Abstract P260: Utilizing Real-World Big Data in the Search for New Renoprotective Drugs
Authors: Horinouchi Yuya , Ikeda Yasumasa , Fukushima Keijo , Imanishi Masaki , Hamano Hirofumi , Izawa-Ishizawa Yuki , Zamami Yoshito , Fujino Hiromichi , Ishizawa Keisuke , Tsuchiya Koichiro , Tamaki Toshiaki
Published: December 2018

Background:
The incidence of chronic kidney disease (CKD) has been increasing globally. Because CKD worsens morbidity and mortality, elucidation of its mechanism and discovery of novel therapeutic strategies are imperative.

Purpose:
We utilized real-world big data to search for potential novel drugs for the treatment of CKD.

Method:
Signal detection by reported odds ratios (RORs) was performed using the Food and Drug Administration Adverse Events Reporting System (FAERS), one of the largest global databases. We then analyzed the relevance between existing drugs and nephritis. Drugs with low reported rates of nephritis were identified as candidate drugs for CKD; their renoprotective effects were verified by basic research.

Results:
Analysis of the FAERS database revealed a significant inverse association between tubulointerstitial nephritis and factor Xa (FXa) inhibitors (ROR, 0.65; 95% CI, 0.49-0.85), and between nephritis and cholinesterase inhibitors (ROR, 0.23; 95% CI, 0.07-0.71).

The FXa inhibitor (FXa-I) suppressed unilateral ureteral obstruction (UUO)-induced tubulointerstitial fibrosis (UUO, 5.20 ± 0.73%; UUO+FXa-I, 3.36 ± 0.32%;
p
< 0.05) and extracellular matrix expression (collagen I, collagen III, and fibronectin;
p
< 0.01). The FXa-I also attenuated UUO-induced macrophage infiltration (UUO, 9.59 ± 0.62%; UUO+FXa-I; 6.70 ± 0.98%;
p
< 0.05) and inflammatory molecule upregulation (monocyte chemoattractant protein 1 [MCP-1], interleukin 1 beta [IL-1β], and tumor necrosis factor alpha [TNF-α];
p
< 0.01). Furthermore, the FXa-I significantly reduced the UUO-induced increase in plasma creatinine in mice (UUO, 0.163 ± 0.005 mg/dL; UUO+FXa-I, 0.136 ± 0.011 mg/dL;
p
< 0.05). The cholinesterase inhibitor significantly mitigated the increased expression of collagen I and MCP-1 in aristolochic acid-induced CKD mice (
p
< 0.05). That also inhibited elevated expression of IL-1β and TNF-α in lipopolysaccharide-stimulated J774 mouse cells.

Conclusion:
We believe that the anti-inflammatory effects of FXa inhibitors and cholinesterase inhibitors make them potentially useful renoprotective drugs. Thus, real-world big data might be utilized for the discovery of drug repositioning candidates for the treatment of CKD.
Abstract P268: Extracellular Vesicles from Wistar Kyoto and Spontaneously Hypertensive Rats Have Differential Vasodilatory Effects on Resistance Arteries
Authors: Erdbruegger Uta , Good Miranda , Musante Luca , LaSalvia Sabrina , Howell Nancy , Carey Robert M , Isakson Brant , Le Thu H
Published: December 2018

Extracellular vesicles (EVs) have been described as novel bio-markers and bio-activators in vascular dysfunction in HTN. However, the exact mechanism how EVs affect vascular function is not known. We hypothesized that hypertensive and normotensive EVs have differential vasodilatory effects on resistance arteries. To examine the effects of EVs on acetylcholine (ACh)-mediated vasodilation, we freshly isolated 3
rd
/4
th
order mesenteric arteries and circulating EVs from 6 and 12-week-old normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats. Arteries were cannulated on a pressure myograph and pressurized to 80 mmHg. EVs (~6x10
7
EV/mL) from hypertensive and normotensive 6 and 12 week old rats were added to the vessel lumen and circulating bath solutions and equilibrated for 10 min. We also added EVs after delipidation with Chloroformmethanol. Inner diameter was measured as cumulative concentrations of ACh were applied to the bath following a preconstruction with 10 μM phenylephrine (PE). No significant difference in ACh vasodilation was observed in arteries from WKY and SHR rats, although SHR arteries were more vasoconstrictive to PE. Interestingly, EVs from WKY had an anti-vasodilatory effect on WKY arteries, whereas EVs from SHR had no effect compared to control condition without EVs. This differential effect was not observed in arteries from SHR rats treated with WKY or SHR EVs. The anti-vasodilatory effect on WKY arteries was also achieved by infusing either delipidated EVs from 12 week-old SHR, or intact EVs from 6 week old SHR rats that have not yet developed hypertension. Together, these data suggest that EVs from normotensive WKY rats and pre-hypertensive SHR rats have anti-vasodilatory effects on healthy resistance arteries. Additionally, the anti-vasodilatory effect of pre-hypertensive SHR EVs is dependent on intact vesicles, but not that of WKY EVs. The effect of normotension on EV cargo might be more important to vasodilatory response than EV species source. This data supports the functional role of EVs in vascular regulation in HTN. Further studies are needed to delineate the effector cargo of these functional EVs.
Abstract P275: Transglutaminase-2 Contributes to Angiotensin II-Induced Reactive Oxygen Species Production in Mice
Authors: Ucci Sara , Chiandotto Sergio , Verga Falzacappa Cecilia , Battistoni Allegra , Volpe Massimo , Savoia Carmine
Published: December 2018

Transglutaminase type II (TG2) is a pleiotropic enzyme that exhibits various activities and it is involved in diverse biological functions, including cell signaling, cytoskeleton rearrangements, displaying enzymatic activities. We previously demonstrated that TG2 may contribute to angiotensin II-induced reduction of NO bioavailability as well as to the impaired vascular functional and structural alterations induced by angiotensin II. Here we hypothesized that TG2 may contribute to increased production of reactive oxygen species (ROS) in the vasculature of angiotensin-II-treated mice. TG2-knockout mice (TG2-K/O, 8 weeks old, n=6) and age-matched wild type (WT) control mice were treated or not with angiotensin-II (400ng/kg/min) for 14 days. TG2 activity in aorta was measured by ELISA. ROS production in aorta was evaluated by dihydroethidium staining. The expression of angiotensin type1 receptor (AT1R), TG2, NOX-1, and ERp72 (the positive modulator of NOX-1) was evaluated in aorta by immunoblotting, coimmunoprecipitation analysis was also performed. As expected, TG2-K/O lacked TG2 expression and activity. Angiotensin-II significantly increased (2-fold) TG2 expression and activity only in WT. AT1R expression in aorta was not influenced by Angiotensin II treatment in both WT and TG2K/O mice. ROS production was similar in WT and TG2-K/O and increased only in angiotensin-II-treated WT (+9%, p<0.01). NOX-1 and ERp72 expression was similar in WT and TG2-K/O. Angiotensin-II significantly increased NOX-1 (+23%, p<0.01) and ERp72 (+29%, p<0.01) only in WT. Only in aorta from WT and not from TG2-K/O, TG2 was successfully immunoprecipitated by AT1 and ERP72, indicating that TG2 is able to interact with both proteins, and suggesting that it may be involved in angiotensin II- induced NOX modulation and ROS production. In conclusion, Angiotensin-II increased ROS production and NOX-1 expression and activation only in presence of TG2 in WT. TG2 interacts with both AT1R and ERp72. Thus, TG2 may contribute to NOX-induced ROS production in mice treated with angiotensin-II.
Abstract P276: Hydrogen Sulfide Regulates Local Blood Flow in Conscious, Free-Moving Rats
Authors: Loredo Juan H , Barrera Adelaeda , Garcia Joshua , Naik Jay , Kanagy Nancy L
Published: December 2018

Inhibiting cystathionine gamma-lyase (CSE) to prevent H
2
S synthesis augments constriction in isolated mesenteric and renal arteries, but H
2
S regulation of resistance in these vascular beds in vivo is unknown. This study evaluated regulation of blood flow and resistance in the mesenteric and renal vascular beds in chronically instrumented rats. Based on studies in isolated arteries, we hypothesized that endogenous production of H
2
S by CSE decreases vascular resistance more in mesenteric than in renal arteries. Rats were anesthetized with inhaled isofluorane (2%) and instrumented with femoral vein catheters, a telemeter blood pressure device and a pulsed Doppler flow probe (Crystal Biotech) on either the superior mesenteric artery or the renal artery (after denervation of the renal artery by removing all visible nerves and painting the surface of the artery with 10% phenol). After five to seven days of recovery from surgery, arterial pressure (AP), heart rate (HR) and relative blood flow (Q) was recorded. Rats then received a daily injection of the irreversible CSE inhibitor, propargylglycine (PAG, 50 mg/kg/day). After five injections, AP was significantly increased in rats with mesenteric flow probes (110±3 start to 122±4 mmHg post PAG, p<0.05 n=7 rats per group). Relative mesenteric resistance (AP/Q) normalized to starting values was increased in PAG rats (150±7% starting value) compared to no change in rats receiving saline (98±3% starting value) (p<0.05, n=7 rats per group). In the rats instrumented with renal flow probes, AP was also increased in the PAG but not in the saline rats (115±6 start to 143±5 mmHg post PAG, p = 0.0014) and calculated renal resistance increased (198% ± 14% versus 103 ± 5% starting value, p=0.0003, n = 5 rats). Heart rate was not changed by any of the treatments. Our results suggest that endogenous H
2
S is an important regulator of vascular resistance in both the mesenteric and renal circulations and that the contribution may be larger in the renal bed than in the mesenteric bed.
Abstract P285: Carotid Sinus Nerve Stimulation Attenuates Alveolar Bone Loss in Rats With Induced Periodontitis.
Authors: Ribeiro Aline B , Fernandes Patricia G , Brognara Fernanda , Castania Jaci A , Silva Carlos A , Messora Michel R , Salgado Helio C
Published: December 2018

Introduction:
Few studies have focused on the impact of the progression of periodontitis on hypertension. Baroreflex Activation Therapy (BAT) has been used to treat patients with resistant hypertension. Our laboratory has investigated the role of electrical carotid sinus nerve (CSN) stimulation in unanesthetized rats, on local and systemic inflammation. Since periodontitis is a chronic inflammatory disease, the aim of this study was to evaluate the role of the baro- and chemoreflex activation, by means of electrical stimulation of the CSN, on alveolar bone loss in rats with periodontitis.

Methods and Results:
Under ketamine/xylazine anesthesia Wistar Hannover rats were implanted with electrodes around the CSN and a catheter inserted into the abdominal aorta for blood pressure recording. After 48h periodontitis was induced by the ligation of the bilateral mandibular first molar, followed by the electrical stimulation of the CSN (1,5-4V, 1ms, 30Hz for 10 min); which was continued during the next eight days. At the 8
th
day after the induction of periodontitis, the rats were euthanized and the jaws were resected; besides, microtomographic analysis was performed by bi and three-dimensional quantification using micro-computerized tomography. As compared to baseline results the electrical stimulation of the CSN (N=7) promoted a decrease in mean arterial pressure (108± 8 vs. 91± 5 mmHg; p<0.05) and heart rate (374±10 vs. 319±15 bpm; p<0.05). The CSN stimulated rats (N=7) with periodontitis showed greater bone volume and bone surface than the non-stimulated control rats (N=4) with periodontitis, evaluated by three-dimensional analysis (0.5±0.005 vs. 0.7±0.03 mm
3
; 16.54±0.22 vs. 23.30±0.66 mm
2
, respectively; p<0.05). Moreover, the CSN stimulated rats with periodontitis presented a decrease of the furcation area and interproximal region as compared to the non-stimulated control rats (1.14±0.06 vs. 0.5±0.07 mm; 3.25±0.03 vs. 2,48±0,13 mm, respectively; p<0.05).

Conclusions:
Overall, these results demonstrate that stimulation of the CSNs promotes a protective effect on alveolar bone loss elicited by periodontitis involving the activation of the baro- and chemoreflex.
Abstract P292: Gene Editing Rat Resource Center: Rat Models for Heart, Lung, Blood, and Sleep Disorder Studies
Authors: Geurts Aron M , Schilling Rebecca , Grzybowski Michael , Temple Anne , Zappa Allison , Lazcares Lynn , Niebuhr Jessica , Kalloway Shawn , Foeckler Jamie , Takizawa Akiko , Dwinell Melinda R
Published: December 2018

Transgenesis and gene editing in the rat has produced gene-modified strains which can be used to validate a gene underlying a quantitative trait locus or human association, or to design follow up studies to define function and physiological roles of genes or sequence variants. We are currently in the fifth year of developing rat models for investigators to validate human variation and study function and mechanism for genes implicated in heart, lung and blood disorders. The Gene Editing Rat Resource Center (GERRC,
http://rgd.mcw.edu/wg/custom_rats/gerrc
) is an NHLBI-funded R24 resource program to generate, distribute, and cryopreserve novel rat models to the research community. Investigator-initiated applications to produce novel genetically modified rat strains were received from laboratories across the world and reviewed by an external advisory board for scientific merit and potential broad interest to the heart, lung, blood, and sleep disorder research community. Custom knockout, knockin, and transgenic rat models continue to be developed. To date, we have transferred over 19,570 microinjected embryos, resulting in 4000+ live-born pups of which 3400+ have been screened for transgenesis or mutagenesis of the target gene. Of these, we have successfully generated ~630 pups containing transgenes or targeted mutations in 95 genes, distributed among 16 inbred, outbred, consomic and congenic rat strains frequently used in cardiovascular research. After confirming germline transmission, heterozygous breeders are distributed to the requesting investigator and then each model is made available to the rat research community. Sperm is cryopreserved to maintain a permanent source of these models. Collectively, the GERRC resource represents the largest collection of genetically modified rat models which are distributable to any investigator through a standard materials transfer agreement at the cost of rearing and shipping. We will discuss the general progress of genetic engineering in rats, current challenges for the field, and opportunities for future developments.
Abstract P294: Intergenerational Obesity: Impact on Cardiac Function and Reserve in Mice Fed a High Fat Diet
Authors: Gava Fabio N , Hall John E , da Silva Alexandre A , do Carmo Jussara M
Published: December 2018

Obesity is associated with structural and functional changes in the heart and abnormal cardiovascular responses to exercise in humans and experimental animals. However, the impact of intergenerational obesity on cardiac function and reserve during increased stress is still unknown. In this study we examined if intergenerational obesity alters cardiac function and responses to a stress test induced by dobutamine in 22-week-old male lean mice fed a control diet (n=5) and obese mice fed a high fat diet (HFD) after birth and that were offspring of mothers who were fed a HFD (F1-HFD, n=7). Mice were instrumented with venous catheters for continuous infusion of saline and progressively increasing doses of dobutamine (2-12 ng/g/min, 2 min at each dose). Long and short axis left ventricle dimensions were obtained before and one minute after each dose of dobutamine using a 30 MHZ transducer (VEVO3100). Baseline heart rate (HR) was similar in F1-HFD (388±15 bpm) and control (426±22 bpm). Compared to controls, F1-HFD mice exhibited impaired diastolic function (E’/A’ ratio: 0.9±0.1 vs. 1.4±0.1 mm/s and isovolumetric relaxation time: 30±2 vs. 21±1 ms) but increased baseline ejection fraction (EF) (84±1 vs. 64±4 %). Dobutamine infusion increased HR by 6±3 and 41±11 bpm, and EF (84±1 to 93±2 and 64±4 to 94±2 %) in F1-HFD and controls, respectively. These results indicate that intergenerational obesity is associated with diastolic dysfunction, but no major alterations in cardiac reserve in response to a cardiovascular stress test. (NHLBI-PO1HL51971, NIGMS P20GM104357 and U54GM115428)
Abstract P299: Evaluation of the Knowledge of Risk Factors Related to Diabetes Mellitus in a Cross High-Risk Population Sample
Authors: Dal Pai Maria Clara D , da Costa Odara , Peçanha Laura M , Saraiva Beatriz G , de Freitas Bernardo P , Wolff Andressa Regina d , Garcia Caroline C , Garcia Renan A , de Castro Nicole C , Brustolin Fernando R , Coimbra Bruno V , da Costa Lilian S
Published: December 2018

Introduction:
Risk factors (RF) for the development of diabetes mellitus (DM) are highly prevalent in high-risk cardiovascular populations.

Objective:
To evaluate the knowledge of RF in the development of DM in a cross-sectional sample of patients attended at an outpatient clinic of a Cardiology Hospital in Rio de Janeiro, Brazil.

Methods:
There were 147 participants (53.7% women), mean age of 63 ± 11.9 years and education level divided into illiteracy (2%), elementary school (44.2%), average (38.1%) and higher school (15.7%). Among the participants, 85% reported hypertension (HY), 25.9% DM and 60.5% previous cardiovascular events, myocardial infarction and/or stroke. Other RF found were: dyslipidemia (46.9%); smoking (10.2%); physical inactivity (66.7%); family history of cardiovascular diseases (52.4%); overweight (25.2%) and obesity (37.4%). A structured questionnaire was applied, evaluating the following question: "In your opinion, what are the FIRST, SECOND and THIRD factor of importance in the development of diabetes?"

Results:
The first, second and third factors of importance were the following distribution, respectively: excessive sugar intake (68.5%, 23.3% and 8.2%), excessive carbohydrate intake (23.3%, 43.4% (2.8%, 10.9% and 29.3%), stress (2.7%, 6.2% and 17.2%), obesity (0.7%, 7% , 8% and 17.2%), smoke (0.7%, 3.1% and 5.2%), dyslipidemia (uncalculated, 4.7% and 1.7%) and HY (uncalculated, 8% and 1.7%). Thus, the eight RF most cited were inadequate food in sugar (100%) and carbohydrates (85.7%), sedentary lifestyle (44.3%), stress (26.1%), obesity (25.7%), smoking (9%), dyslipidemia (6.4%) and HY (2.5%). It should be noted that the excessive intake of sugar and carbohydrate, sedentary lifestyle, stress and obesity were the most frequently cited RF in a population with a high cardiovascular risk, with a high percentage of sedentary lifestyle and overweight. There was no difference in the percentage of knowledge distribution of RF between DM and non-DM, and between HY and non-HY (p = 0.326).

Conclusion:
Although participants have demonstrated knowledge of RF, it is evident the need for better strategies to reduce the discrepancy between the knowledge and the high percentage of RF aggregation in this population.
Abstract P310: Hypermethylation of Dlk1-meg3 Loci in Human Umbilical Veins: Insights Into Offspring Vasculardysfunction Born After Pre-Eclampsia
Authors: Jiang Ying , Yu Yi-Chen , Luo Qiong
Published: December 2018

Increasing epidemiological studies have confirmed the association between maternal pre-eclampsia and elevated blood pressure in their offspring. In our study, we explored the potential epigenetic regulation of Dlk1-Meg3 region in human umbilical vein endothelial cells, and its connection with endothelium-derived factors. We recruited 58 singletons born spontaneously and 67 singletons born with pre-eclampsia during 2016 to detect blood pressure and growth development index, and found that diastolic blood pressure was significantly lower in pre-eclampsia offspring who born over 34 weeks compared to normal offspring (53.59±1.38mmHg VS 59.9±1.40mmHg, P<0.01), which leads to higher pulse pressure difference. RT-qPCR showed that imprinted gene Dlk1 level significantly increased and Meg3 level decreased in HUVECs than that in control group, accompanying with lower expression of endothelial nitric oxide synthase(eNOS) and vascular endothelial growth factor (VEGF) , higher expression of endothelin-1 (ET1), which are close related with vascular endothelial function. Meanwhile, ELISA assay of ET1, Nitrite, VEGF were consistent with Real-time results. Besides, we used pyrosequencing to find that abnormal expression of Dlk1-Meg3 expression were caused by hyper-methylation status of IG-DMR. And methylation status of IG-DMR highly correlated with ET1 concentration and Nitrate concentration, these might be one of the mechanisms for impaired endothelial function (Coeffient=0.5806, P=0.0115; Coeffient=-0.4883, P=0.0398). Our results demonstrated that altered expression of imprinted gene Dlk1 and Meg3 were caused by hypermethylation of IG-DMR in pre-eclampsia HUVECs, accompanied by lower secretion of nitrite, VEGF, and higher secretion of ET1. It might be one potential mechanism for higher risk of cardiovascular disease in pre-eclampsia offspring later in life.
Abstract P314: Regulatory Dendritic Cell Treatment Prevents the Development of Vasopressin-Induced Preeclampsia
Authors: Scroggins Sabrina M , Santillan Donna A , Sandgren Jeremy A , Pierce Gary L , Sigmund Curt D , Grobe Justin L , Santillan Mark K
Published: December 2018

The concept that persistent feto-placental intolerance is important in the pathogenesis of preeclampsia (PE) has been demonstrated by our lab and others. Arginine vasopressin (AVP) infusion during pregnancy induces cardiovascular, renal, and immune alterations in mice consistent with human PE. These findings identify AVP as a potential contributor to poor fetal tolerance and the development of PE. In addition to their conventional immuno-stimulatory role, dendritic cells (DCs) also play a vital role in immune tolerance. In contrast to conventional DCs, regulatory DCs (DCregs) express low levels of co-stimulatory markers, produce anti-inflammatory cytokines, induce T regulatory cells, and promote tolerance. In mice, DCregs are able to prevent pro-inflammatory responses and induce antigen-specific tolerance. Given these known functions of DCregs, we hypothesize that DCregs will prevent the development of AVP-induced PE. C57BL/6J dams were infused with AVP (24 ng/hour) or saline throughout gestation via osmotic minipump. To generate DCregs, bone marrow derived cells from C57BL/6J mice were cultured with human TGF-b1, and murine GM-CSF and IL-10 and phenotype confirmed via flow cytometry. At the time of pump implantation or early post-placentation on gestational day (GD) 7, AVP dams received a single intravenous injection of DCregs. Blood pressure was taken throughout pregnancy and total urine protein was measured on GD 17. Maternal tissues were collected on GD 18. Cytokine concentrations were determined via commercially available ELISAs and normalized to total protein. Treatment of AVP-infused dams with DCregs before mating (GD -3) and on GD 7 prevented AVP-induced hypertension (AVP: 120 ±1.8, n=27 vs GD -3: 108 ±3.3, n=7 vs GD 7: 110 ±4.4, n=5 p<0.05) and elevations in urine protein (AVP: 37.4 ±2.3, n=24 vs GD -3: 25.6 ±2.9, n=7 vs GD 7: 24.1 ±3.1, n=5 p<0.05). Treatment with DCregs also reversed AVP-induced suppression of anti-inflammatory TGFb (AVP: 1.3, n=9 vs GD -3: 3.2, n=7 vs GD 7: 2.9, n=5 ug/g, p<0.05) in the plasma. These data support the hypothesis that DCreg treatment prevents AVP-induced PE. It further provides evidence for the use of personalized, cellular therapy in the prevention of cardiovascular, renal, and immune alterations induced in PE.
Abstract P321: Substance P is Associated with Hypertension-induced Cardiac Fibrosis
Authors: Widiapradja Alexander , Yu Christine , Kott Katharine , Figtree Gemma A , Levick Scott P
Published: December 2018

Myocardial remodelling such as fibrosis is a major contributor to heart failure with preserved ejection fraction. We sought to determine the relationship between substance P (SP), a sensory neuropeptide, and this adverse remodelling. Using the angiotensin II mouse model of pressure overload we determined that

Tac1
-/-

mice that are genetically deficient in SP were protected from developing cardiac fibrosis (WT
+
Ang II = 1.452±0.1523% (n=9) vs

Tac1
-/-
+

Ang II = 0.5678±0.078% (n=10), p<0.0001). We then investigated the association between SP and fibrosis in hypertensive individuals. Plasma from consented individuals showed significantly higher levels of SP in the hypertensive group in comparison to the control group (control = 78.3 ±11.54 ng/mL (n=22), hypertensive = 127±17.5 ng/mL (n=43), p=0.023) (Figure). Interestingly, hypertensive males showed significantly higher levels of plasma SP than the male control group (control = 74.94±14.54 ng/mL (n=17), hypertensive = 143.2±23.96 ng/mL (n=29), p<0.002), while no significant difference existed between these groups in females. Plasma levels of procollagen type 1 C-terminal pro-peptide (PICP), a marker of fibrosis, indicated that hypertensive individuals with SP levels greater than one standard deviation from the mean, had significantly higher levels of PICP (below = 5.539±0.688 ng/mL (n=38), above = 10.52±3.75 ng/mL (n=5), p=0.035). These findings indicate that high levels of plasma SP are associated with increased levels of a marker of fibrosis in hypertensive individuals. The human studies combined with the genetically modified mouse studies indicate an important role for SP in hypertension-induced cardiac fibrosis.
Abstract P323: TMEM16A and TMEM16B Contribute to Mice Cholecystokinin Response in Sex-Specific Manner
Authors: Wang Runping , Cicha Michael Z , Chapleau Mark W , Abboud Francois M
Published: December 2018

High fat diet (HFD) leads to obesity and the metabolic syndrome. However, the effect are less pronounced in females than in males. We suggest that a gender difference is in their response to the satiety peptide Cholecystokinin (CCK). We previously reported that the Ca
2+
-activated Cl
-
channel (CaCC) subunit
Ano2
/TMEM16B is essential for the CCK-induced current in intestinal nodose neurons in male mice, and is down regulated in obese mice fed on HFD. We hypothesize that the CCK-induced current in female mice is mediated predominantly by
Ano1
/TMEM16A, the alternative subunit of TMEM16B. We studied the CCK-induced current in nodose neurons from female C57BL/6 mice using whole cell patch-clamp. We found that 10nM of CCK induces an inward current in these neurons with a current density averaging 13.4±2.5 pA/pF (n=7), and this current is inhibited to 3.1±1.2 pA/pF (n=7, p<0.01) by TMEM16A specific inhibitor T16A
inhi-A01
. We further tested whether HFD which effectively suppressed the expression of TMEM16B in male mice, would also decrease TMEM16A in female mice. Our results show that the CCK-induced TMEM16A current in nodose neurons is not decreased in female, but tend to increase from 12.5±1.6 (n=10) in control to 17.4±4.1 pA/pF (n=11, p=0.298) in female mice fed on HFD. Concurrently the mRNA level of
Ano1
in mice fed on HFD also show a trend to increase by 1.33±0.41 fold (n=2 pairs of mice, p=0.37) compared to mice on regular chow. Our findings indicate that activation of CCK-sensitive
Ano1
/TMEM16A may partially protect female mice from HFD induced obesity and metabolic syndrome.
Abstract P342: Management of Coexistent Neurogenic Orthostatic Hypotension and Supine Hypertension: A Case Study
Authors: Cannom David S
Published: December 2018

A 65-year-old woman presented for evaluation of syncopal spells, up to 5 per week, which typically occurred with assuming the upright posture and on one occasion led to a femoral fracture. Routine orthostatic blood pressure determinations revealed coexistent supine hypertension (SH) with a BP of 150/70 mmHg and neurogenic orthostatic hypotension (nOH), with BP that dropped to 70-80/50 mmHg upon standing associated with a 10-20 beat per minute increase in pulse rate. She was not on diuretics, was euvolemic, and had no evidence of systemic neurological disease or diabetes. A pacemaker was implanted, but it did not resolve her hemodynamic issues and she was admitted to the hospital for medical management of her condition. Losartan was initiated at 50 mg hs and titrated to 100 mg hs to manage the nocturnal SH. Midodrine was started at 2.5 mg TID and titrated to 20 mg TID to manage her nOH. This regimen reduced her orthostatic BP drop by 50 mmHg and relieved her symptoms of syncope, although occasional lightheadedness persisted. As an outpatient, her SH became more severe, reaching levels of 200 mmHg. The dose of midodrine was lowered to 10 mg TID, support stockings were recommended, and clonidine 0.1 to 0.2 mg hs was added to the nighttime regimen. However, this regimen resulted in a recurrence of syncope. She subsequently had a small hemorrhagic stroke, presumably due to the drug-induced hypertension. The patient was readmitted and midodrine was discontinued and replaced with droxidopa, which was titrated up to 300 mg TID. Droxidopa was well tolerated and reduced her orthostatic BP drops by 40 mmHg. She has now been stable with no reported syncopal spells or falls for 3 years. It appeared clinically that droxidopa had a longer half-life, with fewer alarming spikes in her SH than were noted with midodrine. Interestingly, although her presentation suggests OH associated with autonomic dysfunction, she has not demonstrated any primary neurological disease. Midodrine and droxidopa are approved to treat symptoms of orthostatic hypotension and nOH, respectively, and both carry the risk of SH. In this patient with coexisting nOH and severe SH, improvement in nOH symptoms and alleviation of SH were achieved after treatment with droxidopa and a combination of losartan and clonidine hs.
Abstract P346: Impact of the Changing Guidelines in the Classification of Patients: Insights from a Real Life mHealth Digital Registry
Authors: Del Mauro Gaia , Bordignon Tommaso , Fabi Martina , Colombo Eros , Cianflone Domenico
Published: December 2018

Background:
The recent updated AHA/ACC guidelines have spiked academic discussions on the changing prevalence of hypertensive subjects with the new thresholds.

Aim:
To determine the changes in classification (hypertensive or not) on a real life and spontaneous mHealth digital registry, via mobile App and website.

Population and Methods:
7956 consecutive subjects (71% males; 53±11 years old) subscribed, spontaneously, a real-time BP -recording, - interpretation and -trend evaluation service (via smartphone-Health App and website platforms). Thru the digital platforms, each BP value, after consistency-checks, feeds a proprietary algorithm providing both individual BP value interpretation in comparison to the previous BP history, and BP trend and fluctuations assessments. These latter are fed back to users on their smartphone and computer screens instantly. The service complies with the most strict data privacy, safety and security requirements.

Results:
We collected 176432 BP measurements, overall. BP were optimal according to the AHA-ACC guidelines values (<=120/80mmHg) were in 29% of instances, while either only the systolic or the diastolic BP values were optimal in 53% of instances. Conversely, according to the most commonly used normal thresholds of BP <=130/85mmHg 63% of both systolic and diastolic BP values were normal, while either only the systolic or the diastolic BP values were optimal in 39% of instances.

Conclusion:
In a population of subjects, who subscribed a digital remote BP monitoring and evaluation service because they are so aware of the need of controlling their BP, the change of guidelines threshold for optimal and event reducing BP doubles the impact of non-controlled BP. This registry highlights the wide gaps still existing in achieving target blood pressure levels in real-life subjects, even in the most aware hypertensive subjects.
Abstract P350: Early Signs of Stress Induced Arterial Hypertension During Cardiopulmonary Testing and Advantages in Cardiac Risk Lowering
Authors: Kolesnikova Elena A , Arutyunov Gregory P , Rylova Anna
Published: December 2018

Goal:
to study early signs and hypertension reaction features during cardiopulmonary exercise testing (CPET) in intellectually working men with rare episodes of stress induced blood pressure (BP) increase.

Methods:
47 patients (pats) with periodic episodes of high BP due to high level of stress were included (all men, 42 ± 4.6 years old). Risk factors: smoking had 83% of pats, high cholesterol level 87%, glucose intolerance 39%, family history of high BP 14%. All pats had normal BMI and no signs of visceral fat and target organ damage, 54% used hypotensive therapy non-systemically, 98% complained for sleep disorders. CPET was done for optimal training regime appointment. Baseline BP was 134± 2,8 mm Hg. Most of the pats demonstrated low physical tolerance level (6,7±0.4 MET). All patients demonstrated hypertensive reaction during test, and that was the reason for test stop. 68% of pats demonstrated systolic BP 220 mm Hg at respiratory compensation point (RCP) and RER 1,17 ± 0.01, 32% of pats demonstrated it earlier than RCP and at RER 0.85± 0.06. All pats got recommendations with optimal physical and pulse regime for training. All the patients had 3 ± 1 moderate intensive aerobic training per week for 34 ± 10.7 min.

Results:
in 6 months all the patients demonstrated increase in physical tolerance (ΔVO2 5,3 ±1,3 ml/kg/min). Patients with hypertension reaction at RCP showed significantly lower baseline BP (124.6± 3.5mm Hg vs 131.2 ± 4.1 mm Hg in other patients, p <0.05), only 14% demonstrated hypertensive reaction during CPET vs 89% in other patients, p <0.05 and mentioned less hypertension episodes during 6 months (7 ± 3 episodes vs 24 ± 2, p <0.05). 96% pats of this group demonstrated higher level of physical tolerance (load 9,8 ± 0.3 MET vs 7.6± 1.1 MET, p <0,05). Also, they didn't show abnormalities in glucose intolerance test.

Conclusion:
hypertension reaction at RCP during CPET in intellectually working man with episodic stress induced increase of blood pressure can be a predictor of great benefit in life style modification, cardiac risk lowering and arterial hypertension control
Abstract P353: Comparability of Oscillometric to Simultaneous Auscultatory Blood Pressure Measurement in Children
Authors: Ringrose Jennifer S , Alabbas Abdullah , Jalali Afrooz , Khinda Harsimran , Morgan Catherine , Yiu Verna , Alexander R. T , Padwal Raj
Published: December 2018

Objective:
Uncertainty exists regarding the accuracy of automated blood pressure (BP) measurement in children. We recorded oscillometric waveforms in children, derived oscillometric BPs using two standard algorithms, and compared the results to simultaneous auscultation.

Methods:
Twenty children aged 2-12 years were recruited from a tertiary-care Pediatric Nephrology Clinic. Sex, height, weight, arm circumference, history of hypertension and clinic BP were recorded. Two, simultaneously measured, oscillometric and auscultatory BP readings were obtained 30-60 seconds apart. The first reading was discarded and, the second, used for analyses. Fixed-ratio and slope-based algorithms were used for BP derivation.

Results:
Mean age was 7.95 ± 2.82 years, 40% were female, mean arm circumference was 21.86 ± 4.06 cm, and 50% had hypertension or a history of hypertension. Mean auscultatory BP for all participants (systolic ± SD/diastolic ± SD) was 93.40 ± 11.80/50.50 ± 9.04 mmHg, oscillometric fixed-ratio BP was 99.20 ± 11.90/57.35 ± 7.15 mmHg and oscillometric slope-based algorithm was 91.60 ± 13.94/60.65 ± 7.71 mmHg. Compared to auscultation, the fixed-ratio method differed by 5.80 ± 12.72/6.85 ± 7.51 mmHg (p=.06 and p<.01) and the slope-based method differed by -1.80 ± 13.59/10.15 ± 8.07 mmHg (p=0.56 and p<.01). Differences from auscultation were statistically significant for diastolic BP with both fixed-ratio and slope-based methods for all age categories but of greatest magnitude in the youngest children.

Conclusions:
Oscillometric BP derived using two commonly used algorithms differed by more than 5 mmHg in either SBP or DBP from simultaneous auscultatory BP in children aged 2-11. These findings emphasize the need for greater understanding of the functionality and accuracy of oscillometry in children.
Abstract P358: Improvement in Antihypertensive Medication Adherence and Persistence Using a Mobile Technology Application
Authors: McGuiness Catherine , Michaeli Jon , Millard Jennifer , Wang Xin , Wade Rolin L
Published: December 2018

Background:
The high costs and negative outcomes associated with poor medication adherence are well established. This study evaluated antihypertensive adherence and persistence in patients using the Medisafe personalized medication management application (app) compared to a matched control group.

Methods:
Patients installing the app entering an antihypertensive medication (users) between 1/1/2014-12/31/2017 (index date) were anonymously linked to IQVIA’s longitudinal prescription claims (LRx) database and matched 1:3 to non-app users (controls) with prescription claims for antihypertensive therapy in the same month as the app user; further matched on age (+/- 3 years), gender, geographic region, and categorical number of pre-index antihypertensive therapy claims (0-5, 6-11, ≥ 12). All patients had ≥ 2 prescription claims pre-index and ≥ 3 claims post-index, while pre- and post-index periods were equalized for app users and controls. Adherence was measured during the persistent period in LRx via medication possession ratio (MPR) during the pre-index and post-index periods established for both groups. Persistence was defined as no gaps in therapy >60 days. Differences in MPR change between app users and controls were tested using difference in differences (DID) models. Persistence was evaluated using Kaplan-Meyer analysis.

Results:
There were a total of 4,019 cases and 12,057 controls with mean (SD) pre-index periods of 14.2 (8.8) months. The mean (SD) age was 55 (12) years; 55% were male. The net increase in MPR was significantly higher in app-users compared to the controls (+0.083 vs. +0.036, p<.0001) as was the change in the proportion of patients with MPR ≥ 0.80 (+15.40% vs. +7.81%, p<.0001). Persistence on therapy was significantly higher for app users (log-rank test p<0.001) and at 12 months was 81.4% for app users compared to 34.5% for controls.

Conclusions:
Patients using the Medisafe app were significantly more persistent on therapy compared to matched controls and had a higher adherence to therapy during the persistent period. Broad use of this mobile app technology could play an important role in improving clinical and economic outcomes in patients with hypertension.
Abstract P364: Health Insurance Status Affects Hypertension Control in a Hospital Based General Internal Medicine Clinic
Authors: OSO AYODEJI , Adefurin Abiodun , Benneman Monique , Oso Olatunde , Taiwo Muinat , Adebiyi Oluwafisayo , Oluwole Olukemi
Published: December 2018

Objectives:
Hypertension is a worldwide disorder that contributes significantly to morbidity, mortality, and healthcare costs in both developed and developing communities. No previous US study has evaluated the factors associated with blood pressure (BP) control when easy access to health care is available to hypertensive patients.

Methods:
A retrospective cohort study of hypertensive patients attending the Internal Medicine continuity clinic at Nashville General Hospital (NGH) between January and December 2007 was conducted. Given the easy access to health care at NGH and affordable BP medications with a cost of about $4 for a 30 day supply, we explored the ability to achieve optimal guideline directed BP control and evaluated which factors are associated with achieving adequate blood pressure control.

Results:
Of the 199 study participants, 59% of the study population achieved optimal guideline directed BP goal < 140/90mmHg similar to the national average. The mean (95% CI) systolic BP (SBP) was 139 mmHg (136 - 141)mmHg while the mean (95% CI) diastolic BP (DBP) was 80 mmHg (79-82)mmHg. Health insurance status was associated with SBP and DBP (All P < 0.046). Patients with health insurance had a 2.2 fold increased odds of achieving optimal BP control to less than 140/90mmHg compared to patients without health insurance (P=0.025). Furthermore, the number of blood pressure medications used was significantly associated with SBP and DBP (All P < 0.003). Patients taking more than three BP medications had a 58% reduced odds of achieving adequate BP control less than 140/90mmHg compared to patients taking one medication (P = 0.039). Ethnicity was not associated with achieving BP control in this study.

Conclusion:
Our study revealed optimal guideline directed BP control can be achieved when easy access to health care is provided and with use of affordable medications. The number of BP medications used and health insurance status, are factors associated with achieving optimal BP control.