bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 016: Placental Ischemia Exposure leads to a Pro-Inflammatory Environment in Brains of Embryonic Day 19 Rat Offspring
Authors: Warrington Junie P , Logue Omar C , Bidwell Gene L , Shao Qingmei
Published: December 2018

Placental ischemia is thought to be the initiating event in the pathophysiology of preeclampsia, a hypertensive disorder of pregnancy. Previous studies have shown that placental ischemia in the rat, induced by surgically reducing uterine perfusion pressure (RUPP), leads to increased circulating pro-inflammatory cytokines and anti-angiogenic factors, similar to the preeclampsia patient. It is not known whether pups exposed to placental ischemia have evidence of neuroinflammation
in utero
; therefore, this study assessed whether placental ischemia contributes to a pro-inflammatory environment in brains of offspring at embryonic day (E)19. Sham or RUPP surgery was performed on timed-pregnant Sprague Dawley rats (260g) on gestational day 14, and pup brains were extracted on E19. Using a multi-plex cytokine/chemokine kit, we found that 7 out of 27 cytokine/ chemokines were significantly increased in brain homogenates from pups exposed to placental ischemia (n=5 pup brains/group). Higher levels of Eotaxin (5.76±0.26 vs. 3.86±0.84 pg/mg protein; p=0.03), Interleukin (IL)-1β (5.17±0.25 vs. 3.70±0.43 pg/mg; p=0.01), IL-6 (72±10.5 vs. 46.3±7.01 pg/mg; p=0.04), LIX (CXCL5; 45.8±2.2 vs. 30.4±4.7; p=0.01), IL-17 (1.12±0.51 vs. 0.27±0.03; p=0.02), IL-18 (330.1±40.9 vs. 226.5±18.3; p=0.02), and macrophage inflammatory protein 2 (MIP-2; 20.8±0.5 vs. 15.7±1.9 pg/mg; p=0.02) were observed in pup brains exposed to placental ischemia. Despite these increases in pro-inflammatory cytokines, brain water content, measured in a different group of pups from same dams, was not different between the two groups (87.77±0.05 vs. 87.73±0.35%, p=0.53). Whether the increase in pro-inflammatory cytokines in brains of rat pups exposed to placental ischemia is a result of increased transport across the placental barrier or a compensatory mechanism to the hypoxic environment is not known. Our findings support the hypothesis that neuroinflammation in the offspring exposed to placental ischemia may contribute to neurodevelopmental disorders and cognitive impairment observed in offspring of preeclampsia-complicated pregnancies.
Abstract 026: High In-Hospital Systolic Blood Pressure Variability and Poor Functional Outcomes in Primary Intracerebral Hemorrhage Patients
Authors: Meeks Jennifer R , Bambhroliya Arvind B , Meyer Ellie G , Slaughter Kristen B , Fraher Christopher J , Bowry Ritvij , Ahmed Wamda O , Sharrief Anjail Z , Tyson Jon E , Miller Charles C , Khan Babar , Warach Steven , McCullough Louise D , Savitz Sean I , Vahidy Farhaan S
Published: December 2018

Introduction:
High in-hospital SBP variability (HSBPV) is an emerging marker for poor outcomes among Intracerebral Hemorrhage (ICH) patients. We aimed to determine the risk of severe disability or death (SDD) at day-90 among ICH patients with HSPBV and explore pre-hospital factors associated with HSPBV.

Methods:
Adult, radiologically confirmed primary ICH patients were prospectively enrolled and followed-up until day-90. All routinely collected SBP values were recorded for the inpatient stay. Inter and intra-patient SBPV was quantified using generalized estimating equations. Modified Rankin Scale (mRS) Score of 4 - 6 was defined as SDD. Poisson and logistic regression models were fit to determine the risk of day-90 SDD, and the association of pre-hospital characteristics with HSBPV.

Results:
A total of 566 patients [mean age: 63.5, females 36.6% (207 of 566)] were included. Total in-hospital follow-up period was 4,908 days [median (IQR) per patient = 8.7 (3-11)]. Over 120,500 SBP readings were analyzed. Inter and intra-patient mean SBP standard deviation (SD) was 11.1 and 13.2, respectively. A SD of 13.0 was parameterized as a cut-off for HSBPV. HSBPV patients had a 17% higher adjusted risk of day-90 SDD (Relative Risk, 95% CI: 1.17, 1.02-1.35) (Table). Older age and female sex were independently associated with HSBPV after controlling for hemorrhage volume, pre-morbid mRS, and Glasgow Coma Scale (Figure).

Conclusion:
Quantification of HSBPV is feasible utilizing routinely collected SBP readings. HSBPV is associated with poor outcomes. Elderly and female patients may be more likely to demonstrate HSBPV during hospitalization.
Abstract 030: Stem Cell-Derived Extracellular Vesicles Attenuates Aging-Associated Arterial Stiffness and Hypertension
Authors: Feng Rui , Ullah Mujib , Chen Kai , Yi Lin , Lei Han , SUN Zhongjie
Published: December 2018

Objective:
Stem cell-derived extracellular vesicles are decreased with aging. Aging is a major risk factor for the development of arterial stiffness and hypertension. This study was designed to assess the effect of inducible pluripotent stem cells (iPSC)-derived extracellular vesicles (EVs) on the aging-related arterial stiffness and hypertension.

Methods and Results:
Adult (6 months) and aged (20 months) mice were used. Pulse wave velocity (PWV), an index of arterial stiffness and blood pressure were measure twice a week. PWV and blood pressure were increased significantly in aged mice, indicating arterial stiffness and hypertension. One of the exciting findings is that intravenous administration of iPSC-derived EVs significantly decreased aging-related arterial stiffness and hypertension. Treatment with EVs also enhanced endothelium-dependent vascular relaxation and arterial compliance in aged mice. It was noticed that elastin degradation and collagen I deposition (fibrosis) were increased in aortas of aged mice, formulating a structural basis of arterial stiffness. The aging-associated structural remodeling was nearly eliminated by EVs. Mechanistically, EVs abolished downregulation of Sirt1 and eNOS protein expression in aortas of aged mice. In cultured human aortic endothelial cells, we found that EVs promoted the expression and phosphorylation of Sirt1, AMPKα, and eNOS. In addition, EVs attenuated aging-associated renal glomerulosclerosis and tubulointerstitial fibrosis.

Conclusion:
This study demonstrates for the first time that iPSC-derived EVs improved aging-associated vascular endothelial dysfunction, arterial stiffening, renal remodeling and hypertension likely
via
regulating the Sirt1-AMPKα-eNOS pathway. A decrease in stem cell-derived EVs plays an important role in aging-associated arterial stiffening and hypertension. This finding sheds light on the therapeutic potential of EVs for arterial stiffness and hypertension associated with aging.
Abstract 032: Reconstitution of Autophagy Ameliorates Endothelium-Dependent Relaxation, Vascular Smooth Muscle Calcium Sensitization, & Arterial Stiffening in Spontaneously Hypertensive Rats
Authors: McCarthy Cameron G , Wenceslau Camilla F , Calmasini Fabiano B , Klee Nicole S , Webb R. Clinton
Published: December 2018

Insufficient autophagy has been proposed as a mechanism of cellular aging, as this leads to the accumulation of dysfunctional macromolecules and organelles. Premature vascular aging occurs in hypertension. In fact, many factors that contribute to the deterioration of vascular function as we age are accelerated in clinical and experimental hypertension. Previously, we have reported decreased autophagic activity in arteries from spontaneously hypertensive rats (SHR); however, the effects of restoring autophagic activity on blood pressure and vascular function are currently unknown. We hypothesized that reconstitution of arterial autophagy in SHR would decrease blood pressure and improve endothelium-dependent relaxation. We treated 13-18 week old Wistar and SHR with autophagy activator trehalose (2% in drinking water) for 28 days. Blood pressure was measured by telemetry and vascular function was performed on isolated mesenteric resistance arteries using wire and pressure myographs. Treatment with trehalose had no effect on mean arterial pressure in SHR (p>0.05). Trehalose treatment in SHR improved the relaxation to acetylcholine (ACh) [%Relaxation (ACh 1 μM), vehicle: 59.6±8.4 vs. trehalose: 82.4±4.2; p<0.05], and this was not different from arteries incubated with cyclooxygenase inhibitor indomethacin and reactive oxygen species scavenger tempol [%Relaxation (ACh 1 μM), trehalose: 82.4±4.2 vs. vehicle+indomethacin: 92.7±3.9 and vehicle+tempol: 89.3±5.4; both p>0.05]. Trehalose treatment in SHR also prevented the relaxation to Rho kinase inhibition [%Relaxation (Y-27632 0.3 μM), vehicle: 30.4±4.3 vs. trehalose: 8.4±3.5; p<0.05]. Finally, trehalose treatment in SHR decreased arterial stiffness as indicated by the slope of the stress-strain curve [β (stress-strain), vehicle: 8.3±0.6 vs. trehalose: 5.5±0.2; p<0.05]. Overall these data indicate that reconstitution of arterial autophagy in SHR improves endothelial and vascular smooth muscle function, which could synergize to prevent arterial stiffening. As a result, restoration of arterial autophagic activity could be a novel therapeutic for premature vascular aging in hypertension, a recently adopted clinical guideline for cardiovascular disease prevention.
Abstract 036: Interference With PPARγ in the Endothelium Produces Endothelial Dysfunction in the Cerebral Circulation in Response to Activation of the Endogenous Renin-Angiotensin System
Authors: Nair Anand R , Agbor Larry N , Mukohda Masashi , Liu Xuebo , Hu Chunyan , Wu Jing , Sigmund Curt D
Published: December 2018

Low salt diet (LSD) is beneficial in salt-sensitive hypertension but may provoke cardiovascular risk in patients with heart failure, diabetes, or other cardiovascular abnormalities because of renin-angiotensin system (RAS) activation. PPARγ is a transcription factor which promotes an anti-oxidant pathway in the endothelium. We studied transgenic mice expressing a dominant-negative mutation in PPARγ selectively in the endothelium (E-DN) to test the hypothesis that endothelial PPARγ plays a protective role in response to LSD-mediated RAS activation. Plasma renin and angiotensin were significantly and equally increased in all mice fed LSD for 6-weeks (Renin - NT: 39±7 vs 20±1 ng/ml; E-DN: 34±1 vs 16±4 ng/ml; Ang - NT: 257±54 vs 47±6 pg/ml; E-DN: 294±69 vs 63±14 pg/ml p<0.05, n=5). Vasorelaxation to acetylcholine was not affected in basilar artery from E-DN at baseline, but was significantly and selectively impaired in E-DN after LSD (33±5 vs 69±2%, p<0.05, n=6). Unlike basilar artery, LSD was not sufficient to induce vascular dysfunction in carotid artery (carotid artery: 86±4 vs 92±3%, n=5). Endothelial dysfunction in the basilar artery from E-DN mice fed LSD was attenuated by scavengers of superoxide (improved from 29±5% to 55±7%, n=6), inhibitors of NADPH oxidase (improved from 23±3% to 54±7%, p<0.05, n=6), or blockade of the angiotensin-II AT1 receptor (improved from 31±5% to 64±9%, p<0.05, n=5). Gene expression levels of Nox2 was elevated (2.1±0.3 vs 0.4±0.1, p<0.05, n=7) while those of antioxidant enzymes catalase and SOD3 were blunted in cerebral vessels of E-DN mice on a LSD (catalase: 0.5±0.1 vs 2.5±0.2; SOD3: 0.2±0.1 vs 1.1±0.1, p<0.05, n=7). Simultaneous AT1 and AT2 receptor blockade revealed the restoration of endothelial function after AT1 receptor blockade was not a consequence of AT2 receptor activation (59±10 vs 48±2, p<0.05, n=4). We conclude that interference with PPARγ in the endothelium produces endothelial dysfunction in the cerebral circulation in response to LSD-mediated activation of the endogenous RAS, mediated at least in part, through AT1 receptor activation and perturbed redox homeostasis. Moreover, our data suggest that the cerebral circulation may be particularly sensitive to inhibition of PPARγ activity and RAS activation.
Abstract 037: Global ACE2 Over-Expression is Protective Against a Dysfunctional Brain-Gut-Lung Axis in Hypoxia-Induced Pulmonary Hypertension
Authors: Sharma Ravindra K , Oliveira Aline C , Yang Tao , Aquino Victor , Lobaton Gilberto O , Bryant Andrew J , Krause Eric G , de Kloet Annette D , Richards Elaine M , Raizada Mohan K
Published: December 2018

Background and Objectives:
Angiotensin converting enzyme 2 (ACE2), key enzyme of the vasoprotective axis of the renin angiotensin system, has been implicated in many pulmonary diseases including pulmonary hypertension (PH). Plasma levels of ACE2 in group-1 pulmonary hypertensive patients are decreased and activation of this enzyme by small molecule activators, overexpression or administration of recombinant ACE2, all have beneficial outcomes on PH. Furthermore, involvement of neuroinflammation and impaired brain-gut-lung axis have been proposed in PH. These observations have led us to propose that beneficial effect of ACE2 are, at least in part, due to attenuation of neuroinflammation and rebalancing of gut microbiota and improvement in pathology.

Methods:
ACE2 knock-in (ACE2KI) and wild-type mice (WT; C57BL/6) were subjected to hypoxia (10% FIO2) or room air for 4 weeks (n=8-10/group). Right ventricular systolic pressure (RVSP), and electrocardiography were performed, tissues examined by standard histological techniques, and stool collected for 16S rRNA analysis at 4 weeks.

Results:
Hypoxia-induced significantly increased RVSP in WT but not ACE2KI mice (20±1.6 vs 32±1.9, p<0.001 and 24.3±1.5 vs 21±1.3mmHg, respectively). This was accompanied in WT, but not ACEKI, by ~5-fold increase in sympathetic activation (LF/HF; p<0.001) and ~2-fold increase (p<0.01) in microglia activation in the paraventricular nucleus (PVN) of the hypothalamus. There was increased fibrotic area (p<0.01) and muscularis layer thickening (p<0.001) and decreased villi length (p<0.01) and goblet cells (p<0.001) in the small intestine of WT but not ACE2KI mice following hypoxia. Finally, beta diversity of gut microbiome of WT, but not ACE2KI, mice was significantly altered by hypoxia (ANOSIM P=0.001).

Conclusions:
Microglial activation in PVN, sympathetic activation, gut pathology and altered gut microbiome are associated with hypoxia-induced PH. Global overexpression of ACE2 prevents all of these parameters. The involvement of organs other than lungs, and ACE2, present novel therapeutic potentials for PH.
Abstract 046: Cardiac Mitochondria are Impaired After Transient Neonatal High Oxygen Exposure in a Rat Model of Prematurity-Related Condition
Authors: Ravizzoni Dartora Daniela , He Ying , Mian Muhammad Oneeb Rehman , Deprez Alyson , Girard-Bock Camille , Cloutier Anik , Cagnone Gael , Gaub Perrine , Joyal Jean-Sebastien , Luu Thuy Mai , Nuyt Anne Monique
Published: December 2018

The heart relies on adequate mitochondrial (mito) ATP production to match myocardial demand, mostly through oxidative phosphorylation. Preterm birth (PT) results in ex utero development of an immature myocardium and PT-born individuals represent a newly recognized group at high risk of cardiovascular diseases. Our group has shown that newborn rats exposed to high oxygen (O
2
), mimicking PT-related conditions, develop O
2
-induced cardiomyopathy (OIC) and dysfunction later in life. However, whether mito impairments prevail in the programmed left ventricle (LV) changes associated with PT is unknown. We

aimed

to determine whether OIC is associated with altered LV mito characteristics in 4 wks-old rats. Male rat pups were kept with their mother in 80% O
2
(OIC) or room air (Ctrl) from days 3 to 10 of life. Results are mean±SEM; OIC vs. Ctrl are compared using t-test (n=4/group.P<0.05). At 4 wks, extracellular flux analysis of isolated LV cardiomyocytes revealed significantly decreased O
2
consumption rate in OIC (12.14±5.74 vs. 67.39±13.61 pmoles/min/ug). OIC rats show reduced LV mito copy number determined by the ratio between mito and genomic DNA (26.7±6.30 vs. 52.7±7.32), biogenesis markers Pgc1α (0.59±0.14 vs. 1.18±0.25), and citrate synthase (0.20±0.16 vs. 1.60±0.56) mRNA, and reduced Complex IV (0.66±0.15 vs. 1.26±0.09) protein expression. Gene expression (RT-PCR) of key glycolytic enzymes, hexokinase (3.34±0.59 vs. 0.88±0.39) and glucose-6-phosphate dehydrogenase (2.85±0.48 vs. 0.58±0.08) is significantly increased, indicating a shift toward glycolysis. OIC also were found to have higher LV mtROS production (7581±51 vs. 5191±99, fluorescence arbitrary units) and decreased antioxidant SOD2 protein expression (0.84±0.09 vs. 1.04±0.05). Taken together our results indicate that neonatal hyperoxia exposure leads to decreased LV mito biogenesis and function with impaired oxidative phosphorylation capacity in juvenile animals. Further studies are needed to determine the role of long-term mito dysfunction in the increased susceptibility to heart failure observed after deleterious neonatal conditions.
Abstract 048: Impact of New Clinical Practice Guidelines on Cardiovascular Disease Risk Assessment in an Obese Pediatric Population
Authors: Binka Edem , Spevak Philip , Dibert Lauren , McLoughlin Brady Tammy
Published: December 2018

Objectives:
Cardiovascular disease (CVD) risk factor prevalence has increased in children. The new clinical practice guidelines (CPG) revised hypertension (HTN) and left ventricular hypertrophy (LVH) definitions. We investigated the impact of the CPG on CVD risk assessment in obese youth.

Methods:
Cross-sectional study of youth evaluated for high blood pressure at an obesity HTN clinic. Characteristics were compared across historic and current LVH definitions (table 1) using ANOVA. Logistic regression evaluated odds of LVH by HTN status and BMI.

Results:
Of 54 youth, aged 6-21 yrs, 78% (42 of 54) had class III obesity, 57% (31 of 54) had HTN and 17-74% (9-40 of 54) had LVH. Youth with LVH using CPG definitions were older and more obese (table 1). BMI, not HTN, was predictive of LVH using LVMI > 51g/m
2.7
(table 2).

Conclusions:
In this at-risk population, LVH was highly prevalent. Adiposity, not HTN, was predictive of LVH when using the adult definition recommended in the CPG. This may have future implications for recommended CVD risk assessment of youth with severe obesity.
Abstract 050: Hypertensive Urgency: An Emergency Department Pipeline to Primary Care Pilot Study
Authors: Giaimo Antonio , Huot Stephen , Kang Angela
Published: December 2018

Introduction:
The incidence of HTN related ED visits is high and increasing. The optimal management of hypertensive urgency in the ED is not well established but the 2017 ACC/AHA HTN guidelines recommend treatment initiation and outpatient follow up within 1 week. There is a low incidence of poor short-term outcomes associated with hypertensive urgency and high cost but low yield for admitting such patients to the hospital. Our health system lacked an effective referral process for these patients, resulting in poor follow up, unnecessary hospital admission, and recurrent ED utilization. The objectives of our pilot study were to 1) create a process for referring and connecting these patients with outpatient HTN management in a primary care setting and 2) measure the impact of this pipeline referral process on BP control and ED utilization.

Methods:
Patients presenting to the Yale New Haven Hospital ED with hypertensive urgency (SBP ≥180 mm Hg and/or DBP ≥110 mm Hg without end organ damage) who were current patients of the resident physician primary care center or who do not have a PCP were included. Patients meeting inclusion criteria were contacted by a referral coordinator who scheduled their initial appointment to occur within 10 days. Patients were seen by a resident physician supervised by an attending with expertise in HTN. The resident physician leading the visit became their PCP.

Results:
Data are reported for the first 24 referred patients seen in the clinic. Average time to first clinic visit was 7.5 days. Mean age was 54 years (range 32-71), 85% (20/24) were African-American and mean pooled 10 year ASCVD risk was 19%. Mean BP was reduced from 193/111 mmHg at time of ED visit to 131/77 by six weeks after first visit and was sustained at 90 days. Total ED visits by these 24 patients decreased from 30 in the 90 days prior to referral, to 11 in the 90 days after the first clinic visit.

Discussion:
Our results show that ED patients with hypertensive urgency are a high ASCVD risk population. A coordinated referral collaboration between the ED and primary care can provide safe, timely care for this at-risk population that leads to significant reductions in BP and ED utilization. Reduced ED utilization is most likely due to connecting patients with comprehensive primary care.
Abstract 055: ND-13, a DJ-1-Derived Peptide, Protects Against the Renal Fibrosis and Inflammation Associated With Unilateral Ureter Obstruction
Authors: De Miguel Carmen , Saludes Mitchell , Asico Laureano D , Latham Patricia S , Offen Daniel , Jose Pedro A , Cuevas Santiago
Published: December 2018

Oxidative stress and inflammation are important players in the pathogenesis of cardiovascular and renal diseases. DJ-1 is a redox-sensitive chaperone that regulates the expression of several antioxidant genes. Activation of the DJ-1/Nrf2 pathway in the kidney inhibits the development and progression of several renal diseases. The 20 aa peptide ND-13 consists of 13 highly conserved aa from the DJ-1 sequence and a TAT- derived 7 aa sequence to help in cell penetration. ND-13 prevents neuronal degeneration in mice; however, its effects on kidney damage remain unknown. We hypothesized that treatment with ND-13 would prevent the renal damage and inflammation associated with unilateral ureter obstruction (UUO). C57Bl/6 mice and
DJ1

-/-

mice underwent UUO and were divided in 3 groups: control (no UUO), UUO+scrambled peptide (SP) or UUO+ND-13 (3 mg/kg, s.c. daily). After 14 days of treatment, urine and kidneys were collected for analysis of renal damage. ND-13 treatment prevented the development of fibrosis in C57Bl/6 mice (UUO+SP: 702±189% of control, UUO+ND-13: 264±8% of control, n=2-4/group, p<0.05), suggesting that ND-13 is protective against connective tissue deposition in the kidney. Treatment with ND-13 decreased renal mRNA expression of
TNF-
α
(
fold change from control: 101±46 in UUO+SP; 18±7 in UUO+ND-13, n=4-5/group, p<0.05),
IL-6
(6.7±2 in UUO+SP; 1.54±0.3 in UUO+ND-13, p<0.05),
TGF-
β
(
3.3±1.12 UUO+SP; 1.4±0.03 UUO+ND-13, p<0.05) and
Colagen1
α
1
(79±16 UUO+SP; 33±3.8 in UUO +ND-13, p<0.05) in C57Bl/6 mice. In DJ1
-/-
mice, treatment with ND-13 similarly decreased expression of TNF-α, IL-6 and TGF-β, but, in contrast, failed to prevent renal fibrosis or kidney expression of
col1
α
1
. UUO also led to elevated urinary NGAL, marker of proximal tubular injury, in
DJ-1

-/-

mice and ND-13 treatment prevented that increase (71±17% UUO vs control: -18±21% UUO+ND-13 vs control, n=5/group). Our results suggest that ND-13 has protective effects on renal injury, fibrosis and inflammation, crucial mechanisms in the pathogenesis of renal disease. Thus, ND-13 treatment may be a new therapeutic approach for the prevention of renal injury, fibrosis and inflammation in renal diseases. Funded by T32DK007545 to CDM and 5P01 HL074940-10, 7R01 DK039308-31 to PAJ.
Abstract 059: Local Endothelial and Smooth Muscle Genomic Instability Reproduce Specific Features of Cardiovascular Aging
Authors: Bautista-Nino Paula Katherine , Portilla-Fernandez Eliana , Santu Alexandra , Shanahan Catherine , Roks Anton
Published: December 2018

Nuclear and mitochondrial DNA damage contribute to aging and related cardiovascular disease. To explore if the aging features are cell autonomous we compared cardiovascular function of mice with a specific DNA repair system knockout in vascular endothelial cells (EC-KO) to that of mice with specific knockout in smooth muscle cells (SMC-KO). We evaluated cardiac function by echocardiography, blood pressure by the tail cuff method and
ex vivo
vascular function in aorta, coronary and carotid arteries using wire myographs organ baths setups. EC-KO showed macrovascular and microvascular vasodilator dysfunction due to specific loss of endothelium-dependent nitric oxide (NO) signaling (maximal vasorelaxation response to acetylcholine in aorta: 59.0% in EC-KO vs 75.0% in WT mice; p-value lt 0.0001). The reduction of vasodilator response was associated with a temporary systolic blood pressure increase at 3 months (138 mm Hg in EC-KO vs 125 mm Hg in WT mice; p-value= 0.03). In addition, EC-KO mice showed a severely compromised microvascular barrier function in the kidney, leading to papillary necrosis. Cardiac output was slightly affected at 5 months of age (16 ml/min in EC-KO vs 18 ml/min in WT mice; p-value= 0.13) and aortic distensibility was reduced (0.2 mm in EC-KO vs 0.3 mm in WT mice; p-value =0.05), suggesting decreased cardiac contractility and increased vascular stiffness. In sharp contrast, SMC-KO mice showed a specific decrease of endothelium-independent NO-mediated relaxation (maximal vasorelaxation response to the NO donor sodium nitroprusside: 83.4% in SMC-KO vs 98.6% in WT mice; p-value lt 0.001). Furthermore, SMC-KO showed increased carotid artery stiffness (Mediastress at intraluminal pressure of 120 mm Hg: 1.44*10
6
in SMC-KO vs 1.82*10
6
dyness/cm
2
in WT mice; p-value lt 0.001), displayed aortic root dilation and regurgitation. We conclude that DNA damage in EC and VSMC each lead to specific pathological changes that are found also in human cardiovascular aging. In complement, the changes reproduce the vascular phenotype of whole body DNA repair knockout mice (
Ercc1
d/-
) previously found by us. Therefore, the cardiovascular aging effects of DNA damage are at least partly cell autonomic, and represent an important treatment target.
Abstract 061: Mitochondrial Fission Mediates Hypertensive Vascular Remodeling and Aneurysm Development
Authors: Cooper Hannah A , Kawai Tatsuo , Forrester Steven , Elliott Kathy J , Preston Kyle , Scalia Rosario , Rizzo Victor , Eguchi Satoru
Published: December 2018

The prevalence of hypertension is growing steadily with mortality reaching 30,221. While effective blood pressure reduction therapy exists, the underlying pathophysiology leading to vascular remodeling remains poorly understood. Hypertension is a risk factor for abdominal aortic aneurysms (AAA) which when ruptured has 80% mortality. Surgery is the only treatment due to insufficient understanding of the disease process. Mitochondrial dysfunction has been implicated in various cardiovascular diseases but the role of mitochondrial dynamics, a mechanism regulating mitochondrial homeostasis, is under-investigated in hypertension and aneurysm. Our data shows that enhancement of mitochondrial fission via a GTPase, Drp1, in vascular smooth muscle cells (VSMCs) is involved in hypertensive vascular remodeling and aneurysm. In vitro, AngII induced transient mitochondrial fission (2-4 h) and enhanced mitochondrial ROS production in rat aortic VSMCs. Mitochondrial fission, mito-ROS generation, total cell protein, cell volume and extracellular collagen increased by 100 nM AngII were all attenuated in VSMCs by pretreatment with adenovirus encoding Drp1 siRNA/control non-silencing RNA or mdivi1, a Drp1 inhibitor. In vivo, male C57BL/6 mice were infused with AngII (1000ng/kg/min) for 2 weeks (hypertensive remodeling model) +/- mdivi1 (25 mg/kg ip every other day) or infused with AngII for 4 weeks with beta-aminopropionitrile in the drinking water (AAA model) +/- mdivi1 (25 mg/kg ip 3x per week). In the 2-week AngII model, mdivi1 suppressed left ventricular hypertrophy, vascular hypertrophy and perivascular fibrosis induced by AngII in aorta, heart and kidney, independent of blood pressure. In the AAA model, mdivi1 attenuated aneurysm development (External AA diameter (mm) Mean±SEM: 2.15±0.13 vs 1.49±0.07 (, p<0.01). Reduced KDEL and nitro-tyrosine staining in aorta (4w model), coronary and renal arteries (2w) in mdivi1 treated mice, suggests attenuation of ER stress and oxidative stress, respectively. These data suggest that inhibition of mitochondrial fission prevents AngII-induced cardiovascular remodeling and aneurysm development independently of hypertension via ER stress/mito-ROS mechanisms.
Abstract 066: NO Regulates Trafficking of Claudin-19 in the Thick Ascending Limb via c-GMP
Authors: Monzon Casandra M , Garvin Jeffrey L
Published: December 2018

Claudins are a family of tight junction proteins that provide size and charge selectivity to solutes traversing this route. Claudin function can be regulated by posttranslational modifications that affect their localization. Claudin-19 is expressed in TALs and has been implicated in the regulation of Na permeability in this segment. We previously showed that nitric oxide (NO) via cGMP decreases paracellular Na reabsorption in TALs, but the mechanism by which this occurs has not been elucidated. We hypothesize that NO/cGMP regulate claudin-19 trafficking to the plasma membrane. To test this we measured the effects of a NO donor or cGMP on dilution potentials in perfused isolated TALs from Sprague Dawley rats with and without antibodies against an extracellular domain of claudin-19 and Tamm-Horsfall protein (control). Dilution potentials were generated by reducing bath NaCl from 141 to 32 mM. During the control period, the dilution potential was -18.2 ± 1.8 mV. After the NO donor spermine NONOate (SPM; 200 μM) was added, it fell to -14.7 ± 2 mV (n = 8; p < 0.04). In the presence of the claudin-19 antibody, SPM had no significant effect on dilution potentials (claudin-19 antibody alone: -12.7 ± 2.1 mV vs claudin-19 antibody + SPM: -12.9 ± 2.4 mV; n = 6). The claudin-19 antibody alone had no effect on dilution potentials. In the presence of the Tamm-Horsfall protein antibody, SPM reduced the dilution potential from -9.7 ± 1.0 mV to -6.3 ± 1.1 mV (p<0.006, n = 6). Dibutyryl cGMP (500 uM) reduced the dilution potential -19.6 ± 2.6 mV to -17.2 ± 2.3 mV (n = 6; p < 0.02). Dibutyryl-cGMP increased surface expression of claudin-19 from 29.9 ± 3.8% to 65.9 ± 10.1 % (n = 6; p < 0.02) while reducing total expression from 13.3 ± 1.9 to 9.5 ± 1.5 arbitrary units (p< 0.003). We conclude that NO via cGMP reduces Na reabsorption via the paracellular pathway by increasing the amount of claudin-19 in the plasma membrane. We speculate that NO increases both insertion of claudin-19 into and retrieval from the plasma membrane, and the latter leads to enhanced degradation.
Abstract 074: Non-Adherence to Prescribed Blood Pressure Lowering Drugs in Patients With Suspected “Resistant” Hypertension: A Call for Rigorous Adherence Testing
Authors: Ruzicka Marcel , Hiremath Swapnil , Leenen Frans H
Published: December 2018

Background:
Pseudoresistance from non-adherence to blood pressure (BP) lowering drugs can cause unnecessary medical visits and invasive diagnostic tests and procedures. It also leaves patients at a high risk for vascular outcomes. There is currently no unified approach to the diagnosis of non-adherence. Direct questioning by medical personnel and review of pharmacy filling do not reveal the full extent of non-adherence. Direct observational therapy (DOT), a test where a patient’s BP response is monitored after supervised administration of BP lowering drugs, has never been prospectively evaluated for diagnosis of suspected resistant hypertension (HTN).

Methods:
We conducted a prospective study to estimate the prevalence of pseudoresistant HTN using the DOT test. Eligible patients were adults with suspected resistant HTN, defined as daytime average systolic blood pressure (SBP) >135 mmHg on 24-hour ambulatory blood pressure monitoring (ABPM) while on 3 or more BP lowering drugs. Patients confirmed adherence at the clinic, and they had their pharmacy records reviewed. For the DOT, prescribed BP lowering drugs were administered by a nurse at the HTN clinic. BP response was monitored at the office until peak BP effect was reached followed by immediate 24-hour BP ambulatory monitoring, which was repeated at 1 month.

Results:
60 patients were enrolled, and 50 patients completed this study. 30 patients had confirmed resistant HTN. 20 patients had a large SBP decrease during observation immediately post drug administration ( ≥ 20 mmHg) as well as daytime average SBP on 24-hour ABPM (>10 mmHg) as compared to their baseline SBPs. Of these 20 patients, 15 continued to have controlled BP at the 24-hour ABPM done at 1 month post DOT, indicating medium term change in in adherence.

Conclusions:
A large proportion of patients with suspected resistant HTN have pseudoresistant HTN from non-adherence. The diagnosis of pseudoresistance was associated with subsequent cure of resistant HTN in 75% of cases. Proper assessment of adherence is essential to avoid unnecessary drug escalation, and/or investigations for secondary HTN. The DOT represents a simple test for diagnosis and management of suspected resistant HTN, with important ramifications for health care policy makers.
Abstract 077: Captopril-Induced Sustained Reduction in Blood Pressure is Associated With Alterations in Gut-Brain Axis in the Spontaneously Hypertensive Rats
Authors: YANG Tao , Aquino Victor P , Lobaton Gilberto O , Colon-Perez Luis , Zubcevic Jasenka , Febo Marcelo , Richards Elaine M , Pepine Carl J , Raizada Mohan K
Published: December 2018

Objective:
The anti-hypertensive effect of captopril (CAP) is prolonged even when the treatment is discotinued. Our recent findings indicate the beneficial effect of CAP on hypertension-linked gut pathology. Given the roles of brain and gut/microbiota in hypertension, we hypothesized that prolonged anti-hypertensive effect of CAP is through impacts on the brain-gut axis. Accordingly, we tested if CAP induced alterations in the brain and the gut, and if the changes are preserved in the SHR following CAP withdrawal.

Methods:
Male SHR and normotensive Wistar Kyoto (WKY) rats were treated with CAP (250mg/kg/day) in water for 4 weeks followed by its withdrawal for 16 weeks (weeks 5-20). Systolic BP was measured by tailcuff at weeks 3, 5, and 9. Fecal microbiota was analyzed at week 4 and week 8. Brain neuronal activity was measured by manganese-enhanced MRI (MEMRI) at weeks 4 and 8. Gut pathology was assessed at week 20.

Results:
Unweighted principal coordinate analysis showed that CAP treatment significantly altered gut microbial composition, characterized by separate clusters between untreated and 4 weeks CAP-treated SHR. The composition change was preserved 4 weeks post-withdrawal of CAP. Sustained improvement in fibrotic area (untreated: 12.98±0.81% vs week 20: 10.23±0.69%, P=0.036), goblet cell number (untreated: 39±3.57 cells/villi vs week 20: 65±5.29 cells/villi, P=0.0006), and villi length (untreated: 271.4±9.85 μm vs week 20: 388.8±12.29 μm, P<0.0001) was observed at week 20 compared with untreated SHR. MEMRI demonstrated sustained decrease in neuronal activity in autonomic brain regions, with the most pronounced decrease in the posterior pituitary area in SHR treated with CAP (week 4) and withdrawal of CAP (week 8), compared with untreated control (untreated: 1.57±0.11 vs week 4: 1.14±0.074 vs week 8: 1.05±0.14, P=0.0162). In contrast, CAP-treated WKY showed little change in BP, gut pathology or neuronal activity, but distinct changes in gut microbiota.

Conclusion:
Oral CAP persistently lowered BP, altered gut microbiota, improved gut pathology and reduced posterior pituitary neuronal activity in the SHR even post-withdrawal of CAP, indicating persistent antihypertensive effect of CAP may result from altered brain-gut communication.
Abstract 080: Characterization of Blood Pressure Response to Spironolactone in Patients With Resistant Hypertension using Electronic Medical Records
Authors: Shuey Megan M , Nian Hui , Yu Chang , Luther James M , Brown Nancy J
Published: December 2018

Spironolactone is a recommended add-on therapy for BP control in patients with resistant hypertension (RH). We hypothesized that we could use electronic medical records (EMR) to assess the BP response to the addition of spironolactone in patients with apparent RH who were on a stable anti-hypertensive regimen of at least three antihypertensive medications including a thiazide or dihydropyridine CCB.

Patients with RH were identified using a previously published algorithm. We developed an algorithm to identify patients initiated on spironolactone during a period of otherwise stable medication use from up to six months before and after the start of spironolactone to evaluate BP response.

We identified 977 RH patients (751 white and 226 black) prescribed spironolactone during a stable medication window. The median dose of spironolactone prescribed was 25 mg and 724 patients were prescribed 25 mg (74.1%). The mean decrease in SBP following spironolactone was 8.5
+
18.1 mmHg and the mean decrease in DBP was 4.0
+
9.9 mmHg, consistent with data from clinical trials.

Using a mean decrease in SBP of 5 mmHg or in DBP of 2 mmHg to define “responders,” we found that 29% (283 of 977) of patients were nonresponders. Responders had significantly higher baseline BPs (p<0.001), were older (p=0.04), and had larger decreases in the eGFR (p<0.001) and serum Na
+
(p<0.004) and larger increases in creatinine (p<0.001) and K
+
(p<0.001) after starting spironolactone than nonresponders. In blacks, glucose increased following spironolactone in responders compared to non-responders (2.27
+
40.77 vs -15.47
+
62.94 mg/dL, p=0.03); this relationship was not observed in whites.

When response was evaluated as a continuous variable, the decrease in SBP and DBP correlated with the decrease in serum Na
+
(p=0.04) and with the increases in serum K
+
(p=0.01) and creatinine (p<0.001). In blacks, there was a significant correlation between decrease in SBP and increase in glucose (p=0.04).

We have developed an algorithm to assess the BP response to spironolactone in patients with RH using the EMR. Electrolyte changes associated with the BP response to spironolactone are consistent with its mechanism of action to block the mineralocorticoid receptor and decrease activity of the epithelial Na
+
channel.
Abstract 082: Intrarenal Nacl Boli Cause Longlasting Inhibition of Renal Sympathetic Nerve Activity (RSNA)
Authors: Hindermann Martin , Rodionova Kristina , Dietz Amelie , Ditting Tilmann , Ott Christian , Schmieder Roland , Amann Kerstin , Veelken Roland
Published: December 2018

Afferent renal nerve fibers from the kidney likely counterregulate salt sensitive blood pressure increases by decreasing renal sympathetic nerve activity. We recently reported on a long-lasting tonic sympatho-inhibition due to intrarenal afferent renal nerve stimulation eliciting a TRPV1 dependent neuro-humoral pathway. We wanted to test the hypothesis that sodium influences this afferent sympatho-depressory mechanism. Groups of anesthetized SD rats (n=8) were equipped with femoral catheters (blood pressure (BP) & heart rate (HR) recording, drug application), a renal arterial catheter for intrarenal administration (IRA) of high salt (10 % NaCl, 10 μl) or Capsaicin (CAP 3.3, 6.6, 10, 33*10-7 M, 10 μl) and a bipolar electrode for RSNA recordings; eventually an intravenous (iv) bolus of the NK1-receptor blocker RP67580 (10*10-3M, 15 μl) was given. Cultured dorsal root ganglion neurons (Th11-L2) of rats with renal afferents were investigated in current clamp mode to assess action potential generation or in voltage clamp mode to investigate inward currents during 10 sec superperfusion to 4.5 % NaCl or equiosmotic mannitol. solution Results are given in mean±SEM. IRA high salt and IRA CAP decreased RSNA from baseline 4.1±0.6 μV*sec to 2.2±0.8 μV*sec (10% NaCl, p<0.05) and 3.9±0.5 μV*sec to 0.9±0.2 μV*sec (CAP, p<0.01). Suppressed RSNA in high salt groups and CAP could be unmasked by systemic (i.v.) administration of the NK1-blocker (2.7±1.8 μV*sec to 5.8±2.2 μV*sec; p<0.05 (10% NaCl); 1.0±0.2 μV*sec to 6.1±1.5 μV*sec; p<0.01 (CAP)). Cultured renal neurons exhibited production of action potentials (3.5±0.8*, from baseline, p<0.05) and increased sustained inward currents from baseline during exposure to NaCl 4.5 % (-10708.8±3546.5 pA*, from baseline, p<0.05). No responses to equiosmolar mannitol. Increased intrarenal sodium concentrations might induce long-lasting sympatho-depression via a neuro-humoral TRPV1 dependent and tachykinin mediated afferent nerve pathway from the kidney. Impairment of this sympatho-depressory mechanism could be involved in salt sensitive hypertension.
Abstract 087: NPFF Receptors NPFF-R1 and NPFF-R2 in the Kidney: A Target for the Negative Control of Sodium Transport and Blood Pressure
Authors: Asico Laureano D , Villar Van Anthony M , Bishop Michael , Gomes Gerald , Mazhar Momina , Jose Pedro A.
Published: December 2018

The kidney is critical in the overall regulation of fluid, electrolyte balance and blood pressure (BP). The renal dopaminergic system inhibits sodium transport in almost all nephron segments and is responsible for ≥50% of renal sodium excretion under moderate sodium excess conditions and is considered anti-hypertensive. Neuropeptide FF (NPFF), a morphine-modulating peptide, regulates food consumption and cardiovascular function through its interaction with two receptors, NPFFR1 and NPFFR2. NPFFR2 is anti-hypertensive under “normal” salt intake but becomes pro-hypertensive when salt intake is “high”. The ultimate phenotype depends upon protein/protein interaction. Hence, we tested NPFFR2 and dopamine receptor interaction
in vitro
using renal proximal tubule cells (RPTCs) and blood pressure
in vivo
in C57Bl/6J mice. NPFF is synthesized in RPT where NPFFR2 is expressed and co-localize with D
1
R. In RPTCs (n= 3-4/group), the D
1
R/D
5
R agonist fenoldopam (1 μM/30 min), increased cAMP production (4.23±0.56 pmol/mg/min
vs
. 2.54±0.19, vehicle; P<0.05) but was abrogated by co-treatment with NPFF (1 μM/30 (2.36±0.29 pmol/mg/min). Low NPFF (10
-10
and 10
-9
M) concentrations increased while higher concentrations had no effect (10
-8
and 10
-7
M), and slightly decreased (10
-6
M) cAMP production in RPTCs. Fenoldopam (1 μM/30 min) added to the basolateral side of polarized RPTCs grown in Transwells, increased intracellular Na
+
(117±4% versus the vehicle-treated control), indicating inhibition of basolateral Na
+
transport. This effect was prevented by co-treatment with NPFF (98±5%). Basolateral NPFF treatment alone decreased the intracellular Na
+
(-86.10±4.6%), indicating increased basolateral Na
+
transport and that renal NPFF may be a negative regulator of dopamine. In C57Bl/6J mice on 0.9% NaCl diet
, renal-selective silencing of

Npffr2

increased BP (115±3 vs 102±1 mmHg; P<0.05; n=3-5/group). By contrast, on 4% NaCl diet, renal-selective silencing of
Npffr2
decreased BP (98±4 vs 113±3 mmHg; P<0.05; n=3-5/group), indicating that
Npffr2
is antihypertensive on normal NaCl diet but pro-hypertensive on high NaCl diet. These contrasting effects may be due to D
1
R/D
5
R negative regulation of
Npffr2
on normal salt diet which is lost when NaCl intake is increased.
Abstract 090: Does Dietary Sodium Contribute to the Discordance in Blood Pressures among Monozygotic Twins?
Authors: Kidambi Vidya , Donohue Mary F , Munyara Yannick , Coly Gerard , Laud Purushottam , Wang Tao , Reddy Rakesh , Burts S A , Klump Kelly E , Kotchen Theodore A , Cowley Allen W , Rayamajhi Supratik , Mattson David L , Watson Ralph E , Liang Mingyu
Published: December 2018

Background:
Dietary sodium (Na) and central adiposity are considered important risk factors for hypertension (HTN). Blood pressure (BP) levels are discordant among 40-50% of monozygotic (MZ) twin pairs. One prior study of MZ twins using questionnaires reported that hypertensive (HT) MZ twins differed in their intake of meat and milk. However, none of the previous studies have evaluated the impact of Na intake on BP in MZ twins.

Methods:
Eighty-eight pairs of MZ twins were enrolled from Milwaukee, WI and Michigan State University Twin Registry, East Lansing, MI (age- 44 ± 9 years, 63% women, and 96% Caucasian). BPs (measured in triplicate and averaged) and anthropometrics were measured. Na intake was calculated using Block Sodium Screener. Twin pairs were considered concordant or discordant based on systolic (SBP) or diastolic BP (DBP) difference of 10 mm Hg between co-twins, off BP medications

Results:
Thirty-seven twin pairs were discordant in their BP levels. Mean (± SD, mm Hg) SBP/DBPs among concordant and discordant twins were 120 ± 12 / 77 ± 9 and 131 ± 18 / 81 ± 10 respectively. Mean differences in SBP/DBP among concordant and discordant twins were 5 ± 7 / 4 ± 4 and 14 ± 8 / 10 ± 6 respectively. Discordant twins as a group had higher waist circumference (WC) compared to concordant twins (p<0.05), but with no differences in BMI or Na intake. However, among the discordant twin pairs, the co-twin with higher BPs (138 ± 18 / 85 ± 9) had higher dietary salt intakes (3900 ± 1437 vs. 3261 ± 1058 mg/d) and higher WC (105 ± 15 vs. 98 ± 18 cm) (p=0.02) compared to their co-twin with lower BPs (121 ± 16 / 77 ± 9). The average Na intake among the concordant twins was 3443 ± 1189 mg/day. Differences in Na intake among discordant co-twins remained statistically significant after adjustment for age, gender, BMI, WC, and waist-to-hip ratio.

Conclusions:
Higher Na intake is associated with higher BPs among MZ twins with discordant BPs. Differences in Na intakes were significant independent of central adiposity indicating significant Na sensitivity in discordant MZ twin pairs. The effect of ~500mg less Na intake/day in resulting in lower BPs in genetically identical individuals, suggests that modest decreases in Na-intake in Na-sensitive individuals may be sufficient to see the desired effect on BP levels.
Abstract 092: Tissue Sodium is a Highly Sensitive Marker of Subclinical and Localised Edema
Authors: Rossitto Giacomo , Petrie Mark , Touyz Rhian , Delles Christian
Published: December 2018

At variance with the classic physiology concept of constant balance between salt intake and excretion, hypertonic Na
+
accumulation has been shown to occur in peripheral tissues, particularly in the skin. In humans, higher
23
Na MR signal from skin and skeletal muscle has been associated with aging, hypertension, CKD and diabetes.In a rodent model of aging and hypertension, we recently identified an increase in absolute Na
+
content and concentration in the myocardium, but paralleled by water accrual and an associated decrease in K
+
, thus pointing to an overall isotonic accumulation. To further investigate the phenomenon, we developed a mathematical model for expected chemical composition of tissues, as a function of extracellular volume fraction (ECV%) and the amount of fluid isotonic to the extracellular space added to the system (i.e. edema;
fig, closed circles
).Total Na
+
concentration is higher in tissues with larger ECV%; the opposite occurs relative to K
+
(
fig A, open symbols
). The proportional increase in Na
+
concentration due to different degrees of edema is higher than the increase in water content in tissues where ECV% is < 45% (3-4 times higher for skeletal muscle and myocardium, where ECV% is 18-30%;
fig B, black
). When absolute Na
+
content is assessed, similarly to
23
Na MR signal analysis, the proportionally higher increase occurs across the whole spectrum of ECV% (
fig B, red
).We conclude that Na
+
is more sensitive than water to detect edema. This could justify some of the recent evidence of high tissue Na
+
content in multiple conditions, including hypertension, CKD and diabetes, and holds the broad potential for early diagnosis of subclinical congestion and localised edema.