bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P170: Morning Surge and Cardiovascular Events in African Americans: The Jackson Heart Study
Authors: Booth John N , Abdalla Marwah , Sims Mario , Butler Mark , Spruill Tanya , Muntner Paul , Shimbo Daichi
Published: December 2018

Introduction:
A larger sleep-trough morning surge, defined as the mean two-hour post-awakening blood pressure (BP) minus the lowest asleep BP, has been associated with cardiovascular disease (CVD) events in Asians and Europeans. High asleep BP in African Americans (AA) may preclude a morning surge and an association with CVD due to a smaller BP change upon awakening.

Objective:
Determine the association of morning surge with CVD events in AAs.

Methods:
We analyzed participants from the Jackson Heart Study, a community-based cohort of AAs, with a complete 24-hour ambulatory BP monitoring recording and self-report asleep and awake times at baseline in 2000-2004 (n=767). Participants were grouped into tertiles of sleep-trough morning surge, two-hour post-awakening BP and lowest sleep BP. CVD events (nonfatal/fatal stroke, nonfatal myocardial infarction or fatal coronary heart disease; n=61) through December 2012 were adjudicated.

Results:
Participants’ mean age was 59.2 years and 32.1% were male. Multivariable adjusted hazard ratios (95% CI) for CVD events associated with Tertile 2 and 3 versus Tertile 1 of morning systolic BP (SBP) surge were 1.45 (0.71 – 2.97) and 2.03 (0.98 – 4.23), respectively; of post-awakening SBP were 2.01 (0.81 – 4.99) and 4.41 (1.85 – 10.50), respectively; and of lowest asleep SBP were 1.55 (0.68 – 3.52) and 2.22 (0.96 – 5.14), respectively (
Table
). Also, there was a graded increasing risk for CVD associated with Tertile 2 and 3 versus Tertile 1 of post-awakening diastolic BP (DBP), but not morning DBP surge and lowest asleep DBP.

Conclusion:
Higher morning SBP surge, post-awakening SBP and lowest asleep SBP and DBP were associated increased CVD risk in AAs.
Abstract P191: Sexually Dimorphic Metabolic or Lethal Consequences of Disrupting Angiotensinogen in Agouti Related Peptide-Expressing Cells
Authors: Sapouckey Sarah A , Deng Guorui , Pearson Nicole A , Grobe Justin L
Published: December 2018

Genetic disruption of angiotensin (ANG) type 1A receptors (AT1A) in cells which express Agouti-related peptide (AgRP) abolishes thermogenic sympathetic nerve activity (SNA) and resting metabolic rate (RMR) responses to leptin and high fat diet (HFD). Although it is established that glia secrete angiotensinogen (AGT),
in silico
reanalysis of published single-cell RNAseq datasets describing transcriptomes of cells from mouse hypothalamus (GSE74672) and RNAscope fluorescent
in situ
hybridization independently confirm that AgRP neurons of the hypothalamic arcuate nucleus (ARC) in adult wildtype mice express AGT mRNA. We therefore sought to explore the functional significance of this expression pattern. Why, if AgRP neurons are bathed in AGT in the interstitial space, would they synthesize AGT? Mice were generated which lack AGT specifically in AgRP cells (AGT
AgRP-KO
mice) using the Cre-Lox approach (sire: AgRP-Cre
+
, AGT
F/wt
x dam: AgRP-Cre
—
, AGT
F/F
). Of the first 116 offspring, this resulted in expected overall distributions across sex (53% female), Cre (47% Cre
+
) and AGT (46% AGT
F/F
) genotypes, but an underrepresentation of targeted AGT
AgRP-KO
mice at weaning (n=5 females + 1 male; χ
2
=52.0, df=3, p<0.001), indicating pre-weaning or
in utero
lethality of the genotype. Before and at 8 weeks of age, female AGT
AgRP-KO
mice exhibited normal body mass (n=5, 17.3±0.4 vs n=12 littermates 17.3±0.3 g); 5 weeks of 45% HFD caused significant weight gain in all mice (p<0.05), but AGT
AgRP-KO
mice gained less (body mass: 18.3±0.4 vs 19.7±0.3 g, p<0.05, and fat mass by NMR: 1.6±0.4 vs 2.3±0.2 g, p<0.05), which was not due to suppression of food intake (p=NS) or digestive efficiency as assessed by bomb calorimetry (p=NS). We conclude that (i) AGT is expressed by AgRP cells of the ARC, (ii) disruption of AGT in AgRP cells causes a developmental lethality that is more penetrant in males, and (iii) in surviving females, the disruption of AGT in AgRP cells causes resistance to weight gain through increased energy expenditure.
Abstract P195: Blood Presure is Related to Glucose Fluctuation in Longstanding Type 1 Diabetes
Authors: Shalimova Anna , Graff Beata , Szyndler Anna , Wolf Jacek , Blaszkowska Magdalena , Orlowska-Kunikowska Elzbieta , Wolnik Bogumil , Narkiewicz Krzysztof
Published: December 2018

Mechanisms underlying relationship between hypertension and metabolic abnormalities are well established in type 2 diabetes. Much less is known about this link in type 1 diabetes. None of the previous studies has assessed relationship between glucose variability and diurnal blood pressure profile in patients with longstanding type 1 diabetes (DM1).

The Aim:
to investigate the possible association of glucose fluctuation with BP levels in longstanding DM1.

Design and Methods:
We examined 36 patients with longstanding (>20 years) history of DM1 (without overt cardiovascular disease, including hypertension) and episodes of hyperglycemia >160 mg/dL during 24-hour continuous glucose monitoring (CGM). In all patients simultaneous 24-hour CGM and ambulatory blood pressure monitoring (ABPM) were performed. Intima-media thickness (IMT) of the common carotid artery was also assessed. Patients were divided into two groups: with and without severe hypoglycemia <50 mg/dL (n=18 in each group).

Results:
Compared to patients with hypoglycemia, patients without hypoglycemia had a significantly lower day-time SBP variability expressed as standard deviation (12.6 ± 2.5 and 10.9 ± 3.0 mmHg, respectively, p<0.05). In patients without hypoglycemia, mean amplitude of glycemic excursion both up and down was associated with increase in DBP (r=0.49, p<0.05 and r=0.59, p<0.05, respectively), whereas in patients with hypoglycemia it was associated with increase in SBP (r=0.53, p<0.05). In patients without hypoglycemia, time of hyperglycemia was associated with increase in DBP (r=0.57, p<0.05) and in patient with hypoglycemia – with increase in SBP (r=0.67, p<0.05). Furthermore, in patients with short hypoglycemic episodes, time of hypoglycemia was associated with increase in SBP (r=0.57, p<0.05). In patients with hypoglycemia, SBP level had also positive correlation with IMT (r=0.50, p<0.05).

Conclusions:
In patients with longstanding DM1, BP is related to glucose fluctuation. While the change in glucose levels towards hypoglycemia is associated with an increase in SBP, hyperglycemia is linked to an increase in DBP. The mechanisms underlying these associations remain to be determined.
Abstract P199: The Associations Between Blood Pressure Parameters and Stroke in Chinese Rural Population: The Henan Rural Cohort Study
Authors: Zhang Xia , Liu Xiaotian , Li Yuqian , Zhao Xinyu , Liu Min , Tu Runqi , Qian Xinling , Zhang Haiqing , Zhou Wen , Tian Zhongyan , Qiao Dou , Li Linlin , Mao Zhenxing , Zhang Gongyuan , Chongjian Wang
Published: December 2018

Aim:
This study aimed to verify the associations of blood pressure (BP) parameters with the risk of stroke in Chinese rural population. Simultaneously, we quantified how much of the effects of BP on stroke were mediated through metabolic factors.

Methods:
A total of 38880 subjects aged 18 to 79 were derived from the Henan Rural Cohort study. Restricted cubic splines and logistic regression were used to estimate the
OR
s and 95%
CI
s. We used factor analysis to create continuous variables of metabolic dysfunctions to be examined as potential mediator of the association between BP and stroke. Mediation analysis using bootstrap was performed to examine the contribution of metabolic factors to BP parameters related stroke.

Results:
After adjusting for potential confounders, the
OR
s (95%
CI
s) of SBP, DBP, PP and MAP in the highest quartile with the risk of stroke were 1.96 (1.70-2.27), 1.80 1.58-2.05), 1.66 (1.42-1.94), 1.95 (1.70-2.24). The restricted cubic splines showed that the risk of stroke increased gradually with continuous BP levels. We further reported that 11.45%-20.98% of excess relative risk of stroke was mediated through metabolic factors for BP parameters.

Conclusion:
Elevated BP were significantly associated with the risk of stroke especially in SBP and MAP. In addition, metabolic factors explained part of the associations.

Figure 3 Mediation analysis to determine the relationship between SBP (A), DBP (B), PP (C), MAP (D) and stroke through metabolic factors.

M, metabolic factors.OR(95% CI)of total effect for SBP, DBP, PP and MAP on total stroke was 1.0122 (1.0102-1.0142), 1.0205 (1.0169-1.0240), 1.0131 (1.0099-1.0163) and 1.0185 (1.0155-1.0214), respectively.
Abstract P205: Age-dependent Alteration in Expression of Brain-derived Neurotrophic Factor and its Receptors in Central Nervous System of Spontaneously Hypertensive Rats.
Authors: Otani Kosuke , Okada Muneyoshi , Yamawaki Hideyuki
Published: December 2018

Background and Aim:
Essential hypertension is a cause unknown disease leading to heart disease, stroke and renal failure. Spontaneously hypertensive rats (SHR) are animal model of human essential hypertension and develop hypertension at 7-15-week-old. Several studies reported expression pattern of brain-derived neurotrophic factor (BDNF) changes in central nervous system (CNS) in SHR, suggesting that BDNF and its receptors might be related to increased blood pressure. However, age-dependent change in BDNF and receptors (TrkB and p75NTR) remains to be elucidated. This study aimed to clarify it.

Method:
Isolated brain tissues from SHR and control Wistar Kyoto Rats (WKY) at 4-, 7- and 15-week-old were dissected into five regions (cerebral cortex: CC; cerebellum: CB; brainstem: BS; hypothalamus: HT; hippocampus: HC). Protein and mRNA expression of BDNF, TrkB isoforms (FL, T1 and T2) and p75NTR as well as an inflammatory marker NOX-2 was examined by Western blotting and real-time PCR.

Results:
The following data showed significant difference in SHR compared with WKY (n = 4): pro-BDNF protein was lower by 49.9% (4-week,
p
< 0.05) and 38.2% (7-week,
p
< 0.05) in HT, while mature BDNF protein was lower by 21.9% in HC (15-week,
p
< 0.05). TrkB FL mRNA was higher by 32.5% in CB (7-week,
p
< 0.05) and 43.1% in BS (7-week,
p
< 0.05), while TrkB FL protein was higher by 51.4% in BS (15-week,
p
< 0.05). TrkB T1 mRNA was higher by 22.8% in BS (7-week,
p
< 0.05) and 17.2% in HT (15-week,
p
< 0.05), while TrkB T1 protein was higher by 56.1% in BS (4-week,
p
< 0.01). p75NTR protein was higher by 46.9% in CB (7-week,
p
< 0.05). NOX-2 mRNA was higher by 29.1% (4-week,
p
< 0.01), 104.2% (7-week,
p
< 0.01) and 75.8% (15-week,
p
< 0.05) in CC, 27.0% in CB (15-week,
p
< 0.05), 34.6% in BS (4-week,
p
< 0.05), 326.1% in HT (15-week,
p
< 0.01) and 29.8% in HC (7-week,
p
< 0.05).

Conclusion:
We for the first time revealed age-dependent changes in expression of BDNF and its receptors in CNS of SHR. Specifically, our results suggest that alteration of BDNF and TrkB expression is already occurred at juvenile before a stable blood pressure increase. The upregulation of NOX-2 was also identified. Further detailed study might identify whether and how BDNF-receptor-oxidative stress axis is related to pathogenesis of essential hypertension.
Abstract P206: Interarm Difference in Blood Pressure: Prevalence, Risk Factors, and Relevance for Diagnosis of Disease of the Aorta Among Patients Referred to Specialized Regional Hypertension Center
Authors: Jegatheswaran Januvi , Hiremath Swapnil , Edwards Cedric , Ruzicka Marcel
Published: December 2018

Background:
Initial evaluation of hypertension (HTN) should include assessment of blood pressure (BP) in both arms. The prevalence of high interarm differ ranges from 3% in the general adult population to about 10% in the patients with HTN. However, we lack proper estimates of its prevalence, and existing practice of follow up for these patients in a referred population.

Methods:
We performed a retrospective chart review of all prevalent patients followed at the Hypertension Center at the Ottawa Hospital. BP data from the first visit were used for assessment for interarm BP difference. We considered interarm difference in either systolic or diastolic BP in excess of 10 mmHg for casual BP by mercury sphygmomanometry to be clinically significant.

Results:
493 patients of 580 patients were included in this study based on available data. The prevalence of clinically significant interarm difference in systolic or diastolic BP was 16.2% and was similar among men and women. These patients were more likely to be smokers (current or previous; 53.5% vs 36.8%) with peripheral arterial disease (PAD, 15% vs 8%). None of these patients had undergone further investigations of ascending aorta/aortic arch.

Conclusions:
A significant proportion of referred patients have a high interarm difference in systolic or diastolic BP. No clinical investigations were ordered to evaluate for ascending aorta/aortic arch disease reflecting the physicians’ lack of understanding of its clinical relevance. The association with smoking and PAD suggests underlying aortic/large vessel disease as a potential mechanism in some patients.
Abstract P209: Antihypertensive Medication Non-Adherence is Common in Patients With Suspected Refractory Hypertension
Authors: Siddiqui Mohammed , Judd Eric K , Dudenbostel Tanja , Gupta Pankaj , Tomaszewski Maciej , Patel Prashanth , Oparil Suzanne , Calhoun David A
Published: December 2018

Introduction:
Refractory hypertension is a phenotype of antihypertensive treatment failure defined as uncontrolled BP (> 135/85 mmHg) despite the use of five or more different antihypertensive agents, including long-acting thiazide like diuretic (chlorthalidone) and mineralocorticoid receptor antagonist (Spironolactone or Eplerenone). Recently white coat effect has been shown to be uncommon in patients with refractory hypertension. However, the degree of medication non-adherence, an important cause of apparent refractory hypertension, is unknown.

Methods:
In this prospective evaluation, 37 refractory hypertensive patients were recruited from the University of Alabama at Birmingham Hypertension Clinic after having uncontrolled BP at three or more clinic visits. All patients were evaluated by automated office BP (AOBP) with the BpTRU device, ambulatory BP monitoring (ABPM), and 24-hr urine collection to detect antihypertensive medication adherence by high-performance liquid chromatography-tandem mass spectrometry. Out of 34 patients who underwent ABPM monitoring, 30 (88%) patients were refractory hypertensive by AOBP and by ABPM. Of these, 27 patients had 24-hr urine collection for detection of prescribed antihypertensive medications or their metabolites.

Results:
Out of 27 patients, 11 (40.7%) patients had complete adherence with five or more antihypertensive medications, indicating true refractory hypertension. Ten (37.0%) patients were partially adherent, and 6 (22.2%) patients had total non-adherence to their antihypertensive medications.

Conclusion:
Of patients identified as having apparent refractory hypertension, 40% were adherent with at least 5 of their prescribed antihypertensive agents, confirming true refractory hypertension. The remaining 60% were partially or fully non-adherent. These results indicate that non-adherence is common in patients suspected of having refractory hypertension.
Abstract P216: The Circadian Clock Protein BMAL1 in the Kidney Contributes to Blood Pressure Control in Male Mice
Authors: Crislip Gene R , Douma Lauren G , Masten Sarah , Cheng Kit-Yan , Lynch I. J , Holzworth Meaghan , Barral Dominique , Miller Amber , Esser Karyn A , Wingo Charles S , Gumz Michelle L
Published: December 2018

Blood pressure (BP) regulation is influenced by the circadian clock as seen by its daily rhythm, with a peak occurring in the active phase and a dip during the rest phase. BMAL1 is a core circadian transcription factor which regulates the circadian clock feedback loop. BMAL1 also controls the expression of thousands of genes important for physiological functions. Others have shown that global BMAL1 knockout male mice have lower mean arterial pressure (MAP) and a loss of circadian rhythm compared to controls. Because the kidney is a critical regulator of BP, the goal of this study was to determine the BP phenotype of renal distal nephron-specific BMAL1 knockout mice (KO) vs. littermate controls (CNTL). KO were generated using Ksp-cadherin Cre. Histological analysis validated the BMAL1 KO model. MAP was measured by radiotelemetry. KO had 7 mmHg lower MAP compared to CNTL (103±2 vs. 110±0.2; P=0.03; N=4-5), however, the circadian rhythm of BP was apparent in both groups. There was no difference in activity or heart rate (P=0.7, P=0.1). Kidneys were collected at noon (midpoint of rest phase) for gene expression analysis, Table 1. HKa2 expression was increased in untreated KO vs. CNTL (1.7±0.2 vs. 1±0.1; P=0.01). We have previously linked HKa2 to renal Na handling and the mineralocorticoid response. Therefore, we assessed the BP response to a high salt diet plus mineralocorticoid treatment (HS/DOCP). MAP increased in both groups in response to HS/DOCP (P=0.03) where KO rose 6 mmHg (109±1) and WT only 2 mmHg (112±2). These results suggest that BMAL1 in the kidney plays a critical role in the regulation of BP but does not appear to contribute to the circadian rhythm of BP under the conditions tested.
Abstract P224: Urinary Excretion of Sodium, Potassium, Calcium and Magnesium and Blood Pressure Among a Population of ≥ 20-Year-Olds: Evidence From Southwest Coastal Bangladesh
Authors: Titu Abu Mohd Naser , Rahman Mahbubur , Unicomb Leanne , Doza Solaiman , Ahmed Kazi Matin Uddin , Selim Shahjada , Anand Shuchi , Chang Howard , Luby Stephen , Clasen Thomas , Gribble Matthew , Narayan KM Venkat
Published: December 2018

We analyzed data from a stepped wedge randomized trial to evaluate how urinary Na, K, Ca and Mg excretion are associated with the change in blood pressure (BP) among the adult population. We followed up a cohort of 1,191 participants (>20 years old) from 540 households in 16 communities of southwest coastal Bangladesh for five visits during December 2016 - April 2017 when they were exposed to high salinity drinking water. In all visits, we measured participants’ BP (N=5,746) and 24-hour urinary Na, K, Ca and Mg. We used multilevel linear regression models to determine the association among change in urinary excretion of Na, K, Ca and Mg with differences in mean systolic blood pressure (SBP) and diastolic blood pressure (DBP). Models included participant-, household-, and community-level random intercepts and were adjusted for age, sex, BMI, physical activity, smoking status, and household wealth. We restricted analysis among participants who had complete 24-hour urine collection based on measured versus expected urinary creatinine ratio of >=0.7 (N=5,103). The median urinary 24 hours Na excretion was 164 mmol, K was 34 mmol, Ca was 126 mg, and Mg was 83 mg in visit 1. The mean population SBP was 115.6 mmHg in visit 1 and 110.2 in visit 5, and the mean DBP was 68.9 mmHg in visit 1 and 65.9 in visit 5. Compared to visit 1, we found 85% higher urinary Ca in visit 3 and 57% higher urinary Mg in visit 5. We found 100 mmol per 24 hours increase in urinary Na was associated with 1.74 (95% CI: 0.85, 2.62) mmHg higher SBP and 0.51 (95% CI: -0.06, 1.08) mmHg higher DBP, and 50 mmol per 24 hours increase in urinary K was associated with 2.39 (95% CI: 0.84, 3.94) mmHg lower SBP and 0.79 (95% CI: -1.76, 0.17) mmHg lower DBP. We found 100 mg per 24 hours increase in urinary Ca was associated with 0.29 (95% CI: 0.02, 0.60) mmHg lower SBP and 0.32 (0.07, 0.57) mmHg lower DBP, and 100 mg per 24 hours increase in urinary Mg was associated with 1.09 (95% CI: 0.55, 1.64) mmHg lower SBP and 0.41 (0.07, 0.76) mmHg lower DBP. We found SBP lowering effect of urinary Mg increased if urinary Ca was lower (p = 0.048). We found urinary K, Ca, and Mg are associated with lower BP in coastal Bangladesh. Our results suggest a high intake of these beneficial minerals alongside Na reduction may successfully lower mean BP of the population.
Abstract P225: Associations Between Sleep, Physical Activity and Blood Pressure Among Black Women: Baseline Results From the
SisterTalk At Home
Intervention for Weight Loss
Authors: Kang Augustine W , Risica Patricia M
Published: December 2018

Significance:
Sleep is a key factor affecting hypertension and obesity, but limited evidence is available examining the relationship between sleep and predictors of hypertension/obesity (e.g. physical activity (PA)) among populations at risk for health disparities. This study seeks to fill gaps in research by (a) describing sleep patterns among overweight/obese Black women, (b) examining the association between sleep and PA, and (c) examining the association between PA and blood pressure (BP).

Methods:
We examine baseline data from the
SisterTalk at Home
study, an RCT assessing the efficacy of a church-based weight management program for Black women in Northeastern USA. The study sample included 361 participants (Age = 49.9 ± 13.0 years). Mean BMI was 34.8 ± 6.7 kg/m
2
and mean systolic/diastolic BP (SBP and DBP) were 125.9 ± 17.8 and 77.9 ± 9.2 mm/Hg. Study instruments include the Epworth Sleepiness Scale (ESS), self-reported sleep duration, and PA. Chi-square, ANCOVA and linear regression were used for data analysis.

Results:
49% of the sample reported short sleep duration (SSD) (<7 hrs); 46% were optimal sleepers (7-9 hrs), and 5% were long sleepers (>9 hrs). For sleepiness (ESS), 38% of the sample were classified as “normal”, 35% were “high normal”, and 27% had “excessive daytime sleepiness”. Age, unemployment, and BMI were associated with higher SBP (
p
<.001, .006, and .002) and thus controlled for in subsequent analysis. Participants in the “high normal” ESS category reported significantly lower vigorous and moderate PA compared to “normal’ participants (
p
= .028 and .035). Vigorous PA was in turn inversely associated with SBP and DBP (
p
= .009 and .031); Moderate PA was inversely associated with SBP (
p
= .039).

Conclusions:
The present sample reports a higher prevalence of short sleepers and excessive daytime sleepiness compared to the general population, supporting prior findings that poor sleep quality may be comorbid with obesity. Results also indicate a relationship between poor sleep and lower physical activity, suggesting that intervention efforts to address obesity among this population should consider targeting sleep. Lastly, PA was associated with BP, extending conclusions drawn in other populations to our sample.
Abstract P238: Cerebral Ischemia With Amyloid-Beta Infusion Deteriorates Cognitive Decline ~Possible Amelioration of Cognitive Function by AT
2
Receptor Activation~
Authors: Min Li-Juan , Mogi Masaki , Bai Hui-Yu , Shan Bao-Shuai , Higaki Akinori , Iwanami Jun , Horiuchi Masatsugu
Published: December 2018

Objectives:
Amyloid-β (Aβ) deposition in brain and cerebral vessels accelerates the pathogenesis of Alzheimer’s disease and vascular cognitive impairment. We examined possibilities that cerebral ischemia worsens Aβ infusion-mediated cognitive decline, and that angiotensin II type 2 (AT
2
) receptor stimulation in vascular smooth muscle cells (VSMC) could ameliorate this cognitive impairment induced by cerebral ischemia and Aβ.

Methods:
Adult male wild-type mice (WT) and the mice with VSMC-specific AT
2
receptor overexpression (smAT
2
) were used. Mice were subjected to intracerebroventricular (ICV) injection of Aβ1-40. Cerebral ischemia was induced by 15 minutes of bilateral common carotid artery occlusion (BCCAO) 24 hours after Aβ injection. Cognitive function was evaluated by Morris water maze test 3 weeks after Aβ injection.

Results:
ICV injection of Aβ in WT showed impaired cognitive function (arriving time to platform at day 5: control, 26.53±4.46 sec; Aβ, 65.35±7.44 sec), whereas BCCAO did not decline significantly cognitive function. In contrast, BCCAO following Aβ injection exhibited more marked cognitive impairment (84.27±8.00 sec) compared to Aβ injection alone in concert with the increases in superoxide anion production, NADPH oxidase activity, expressions of NADPH oxidase subunit p
22
phox
, p
40
phox
and inflammatory cytokines such as MCP-1, IL1-β in the hippocampus. BCCAO following Aβ injection significantly enhanced the expression of Aβ clearance factor, RAGE (receptor for advanced glycation end product). Aβ injection did not increase the neuron pyknosis in the hippocampus, whereas the number of neuron pyknosis was increased significantly with BCCAO (control, 6.33±0.88/field; Aβ with BCCAO, 46.33±4.10/field). On the other hand, smAT
2
did not show cognitive impairment, the increases in oxidative stress, inflammation markers and RAGE expression, pyknosis, which were induced by Aβ injection with/without BCCAO in WT.

Conclusion:
Cerebral ischemia exaggerated Aβ-induced cognitive decline with possible involvements of enhanced oxidative stress, inflammation, neuronal degeneration, and breakdown of RAGE-mediated Aβ clearance. AT
2
receptor activation in VSMC could play preventive roles in this cognitive decline.
Abstract P240: Identifying the Angiotensin AT2-Receptor Coupled Phosphoproteome in Human Aortic Endothelial Cells by Time-Resolved, Quantitative Phosphoproteomics
Authors: Peluso Augusto B , Kempf Stefan J , Verano Brega Thiago , Bader Michael , Larsen Martin R , Santos Robson A , Steckelings Ulrike M
Published: December 2018

Objectives:
The angiotensin AT2-receptor (AT2R) is a main component of the protective arm of the renin-angiotensin system. In order to get new and deeper insights into AT2R mediated signalling, this study mapped the AT2R-coupled phosphoproteome in human aortic endothelial cells (HAEC) by quantitative phosphoproteomics.

Methods:
HAEC were stimulated with the AT2R agonist C21 (1 μM) for 1, 3, 5 or 20 minutes. Following protein digestion and TMT peptide labelling, samples were subjected to TiO2–SIMAC phospho-peptide enrichment and LC–MS/MS. Data were analysed using Proteome Discoverer and the human sequence libraries SwissProt and UniProt. For confirmation of phosphoproteomics results, expression of p53 and phosphorylation of HDAC-1 were determined by Western Blot. Cellular translocation of HDAC-1 and p53 was assessed by immunofluorescence and activity of HDAC-1 by deacetylation assay. HAEC proliferation was determined by resazurin assay and apoptosis by caspase 3/7 assay.

Results:
AT2R stimulation of HAEC changed the phosphorylation status of 265 proteins at 471 sites (281 phosphorylations and 190 dephosphorylations). Gene Ontology and STRING analysis revealed the predominant phospho-modification of proteins involved in anti-proliferation, cell differentiation and apoptosis including HDAC-1, p53, BCL2 or HSP90. AT2R-mediated inhibition of HDAC-1 and activation of p53 were confirmed by additional experiments demonstrating changes in cellular localisation of HDAC-1 and p53, in total expression of p53, and in enzymatic activity and phosphorylation of HDAC1.

Conclusion:
This study mapped the AT2R coupled phospho-signalling network. It identified dephosphorylation and deactivation of HDAC-1 as well as nuclear importation and increased expression of p53 as novel, potential mediators of AT2R-coupled anti-proliferation.
Abstract P245: The Angiotensin-(1-7) Agonist AVE0991 Attenuates Angiotensinogen Release from Human Proximal Epithelial Cells
Authors: Chappell Mark C
Published: December 2018

A local renin-angiotensin system (RAS) is evident within the renal proximal tubules and overexpression of the precursor angiotensinogen targeted to the tubules increases blood pressure and promotes renal injury. Tubular angiotensinogen is enhanced by the Ang II-AT1 receptor axis, oxidative stress and pro-inflammatory cytokines (IL-6, TGFβ), as well as hyperglycemic conditions. Stimulation of angiotensinogen by the Ang II-AT1 receptor pathway constitutes a positive-feedback loop that may promote a sustained increase in blood pressure and renal damage. Mediators that promote angiotensinogen also attenuate the expression of the ACE2-Ang-(1-7) axis. Downregulation of the Ang-(1-7) axis and its associated signaling pathways may contribute to greater expression of angiotensinogen within the kidney. To examine this potential pathway, we assessed the chronic effects of the Ang-(1-7) agonist AVE0991 on angiotensinogen release in human HK-2 proximal epithelial cells maintained under mild hyperglycemic conditions. HK-2 cells were placed in serum-free media and then treated with varying doses of the AVE0991 agonist (0.1 nM to 1 uM) for up to 48 hours. Human angiotensinogen was quantified in the cell media by ELISA with a sensitivity of 30 picogram (pg). Basal release of angiotensinogen at 48 hrs was 1.47 ± 0.28 ng/ml (n=4). Addition of 0.1 nM AVE0991 attenuated angiotensinogen release by 23 ± 4%, p<0.05 with a maximal inhibitory effect of 48 ± 9%, p<0.05 [n=3]. AVE0991 at the maximal dose had no detrimental effect on cell viability. The NOS inhibitor LNAME (100 uM) failed to block the inhibitory effect of AVE0991 and LNAME alone did not influence release. Addition of the AT2R agonist C21 (1 uM) had no effect on angiotensinogen release. The Ang-(1-7) receptor antagonist [D-Pro
7
]-Ang-(1-7) blocked the inhibitory effects of AVE0991; however, [D-Ala
7
-Ang-(1-7)(A779), losartan, PD123319 or the kinin B2 receptor HOE140 had no effect. We conclude that the Ang-(1-7) axis may exert negative feedback on the tubular angiotensin system and that the renoprotective effects of AVE0991 may reflect attenuation of angiotensinogen release. Furthermore, the inhibitory actions of AVE0091 on the tubular release of angiotensinogen appear to be independent of the NOS-NO pathway.
Abstract P267: Aging & Endothelial Cell Senescence: Does Prolylcarboxypeptidase Predict Endothelial Dysfunction?
Authors: Shariat-Madar Zia , Boullard Nicholas , Cissom Cody
Published: December 2018

Changes due to cellular senescence may lead to the formation of a unique cellular morphology such as the transformation of endothelium from an anticoagulant to a potentially prothrombotic surface. Investigations were performed to determine whether the ratio of prolylcarboxypeptidase (PRCP) gene expression to the rate of telomere shortening in vascular endothelial cells would predict endothelial dysfunction and subsequent endothelial surface alteration from anticoagulant to potentially prothrombotic surface. Here, we present evidence for a novel mechanism of senescence-induced cell-surface expression of PRCP involving activation of the plasma kallikrein-kinin system (KKS). We found that the abilities of human pulmonary vein endothelial cells (HPVECs) to express PRCP first incrementally increased at early passages and then decreased after passage 18, in contrast to changes in average human telomerase reverse transcriptase (hTERT) mRNA expression levels. In agreement with the changes in average PRCP expression, incubation of prekallikrein (PK) and high molecular weight kininogen (HK) plasma proteins resulted in increased cleavage of HK, higher enzymatic activities of kallikrein, and increased liberation of bradykinin (BK). Furthermore, the alterations in PK and BK generation was prevented using PRCP-siRNA or PRCP inhibitor (z-Pro- prolinal) that blocked PK activation in passage 18 by 60% and 80% respectively. SDD31 that interferes with the binding of PK on HK reduced kallikrein generation by 90%. There was a direct correlation between PRCP-dependent BK generation and cell permeability. Our results were supported when we found that nitric oxide (NO), a downstream product of the PRCP-catalyzed HK-PK system, also shared the same peak in formation as PRCP. In contrast, increased eNOS expression remained unchanged after passage 12. While β-Gal rose after passage 18, calnexin expression like PRCP dropped significantly in passage 30. These data are the first to demonstrate the inverse relationship between hTERT and PRCP expression prior to an apparent onset of EC replicative senescence, which could have profound implications as a prognostic tool for predicting disease progression and cardiovascular event rates.
Abstract P269: Chronic Cerebral Ischemic Injury Worsens Vascular Dysfunction
Authors: Iwanami Jun , Mogi Masaki , Tsukuda Kana , Higaki Akinori , Kawakami Moe , Kukida Masanori , Nakaoka Hirotomo , Yamauchi Toshifumi , Bai Hui-Yu , Shan Bao-Shuai , Min Li-Juan , Horiuchi Masatsugu
Published: December 2018

Introduction:
Distant organ dysfunction has been highlighted whereby acute or chronic damage in one organ may induce acute or chronic dysfunctions of the other organs such as cardiorenal syndrome. Vascular remodeling is an important process in various vascular diseases. However, the relationship cerebral ischemic damage and peripheral vascular disorder is still unclear. Therefore, we have investigated the possible pathophysiological interactions between brain ischemia and vascular remodeling.

Methods:
Eight-week-old male C57BL/6 mice underwent permanent cerebral brain ischemia by permanent occlusion of the left middle cerebral artery occlusion (MCAO) by electrocoagulation using a subtemporal approach. Four weeks after MCAO, vascular injury was induced by polyethylene cuff placement on the femoral artery. Neointima formation was determined 14 days after cuff placement by evaluating intima/media ratio. Macrophage fractions in spleen two weeks after MCAO were examined and M1 and M2-polarized macrophages were separated by flow cytometry and analyzed.

Results:
Body weight after MCAO and cuff placement did not differ among all groups. Cerebral ischemia did not influence the intima/media ratio 6 weeks after MCAO (MCAO-Cuff (-), 0.123) compared with sham-operated mice (sham-Cuff (-), 0.120). Neointima formation in the injured artery was significantly increased 2 weeks after cuff placement (sham-Cuff (+), 0.258). Interestingly, the neointima formation 2 weeks after cuff placement was more marked in MCAO-operated mice (MCAO-Cuff (+), 0.369) compared with sham-Cuff (+). Next, we examined the macrophage fractions in spleen 2 weeks after MCAO. The macrophage fraction in SVF evaluated by F4/80 staining did not differ between sham- (7.5%) and MCAO-operated (7.6%) mice. There was no significant difference in M1 and M2 fraction with/without MCAO.

Conclusion:
These results suggest that cerebral ischemic injury aggravate peripheral vascular disorders, and we are addressing the possible roles of renin-angiotensin system and oxidative stress etc. involved in these interesting results, which could provide us with the discovery of new pathological mechanisms concerning the distant organ dysfunctions between brain and peripheral vasculature.
Abstract P271: Relationship of Resting Heart Rate and Subclinical Arteriosclerosis Markers in Apparently Normal Participants From China
Authors: Wu Xiuqin , Hong Yongqiang , Liu Huan , Wang Hongyu , Zhao Hongwei
Published: December 2018

Objective:
To evaluate the relationship between resting heart rate and arterial stiffness in natural and apparently normal population of She minority from Fujian province in China.

Method:
6 natural village 511 participants were enrolled into our study using cluster sampling method in 2009. Eventually 325 subjects with full data were analyzed. All subjects carried on detection of carotid femoral pulse wave velocity(CF-PWV), carotid radial pulse wave velocity (CR-PWV), cardio ankle vascular index(CAVI), ankle brachial index(ABI), carotid intima media thickness(CIMT) by Doppler ultrasound and blood test (fasting plasma glucose[FPG], blood lipids, blood uric acid(UA), high sensitive C reactive protein(hs-CRP). Resting heart rate(RHR) was the mean of three values measured by VS-1000 vascular detection equipment and ultrasound equipment.

Results:
In different quartiles of RHR, that were RHR1≤61beats/min, RHR2:62-69beats/min, RHR3:70-77beats/min, RHR4≥78beats/min, systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol to high density lipoprotein cholesterol ratio (TC/HDL-C), HDL-C, CF-PWV and CR-PWV were significantly different between RHR groups (p<0.05); SBP, DBP, TC/HDL-C, CF-PWV and CR-PWV in RHR4 group were significantly higher than RHR1 group, and HDL-C was lower than RHR1 group (all p<0.05). However, there were no statistical significance of CAVI, ABI and CIMT in different RHR groups (p>0.05). Logistic regression analysis indicated in the model without adjustment, the RHR4 was associated with higher CF-PWV(CF-PWV>9m/s) with odds ratios (ORs) was 3.074 (95% confidence interval was 1.555-6.075, p<0.05) compared with RHR1; After adjustment of age and gender, the OR of RHR4 was 4.542 (95% confidence interval was 2.078-9.928, p<0.05); And after further adjustment of SBP, DBP, plasma lipids, FPG and hs-CRP, the OR of RHR4 was 5.336 (95% confidence interval was 1.512-18.831, p<0.05).

Conclusion:
In natural population of She minority of Fujian province in China, high resting heart rate was followed by a high level of blood pressure, plasma lipids and arterial stiffness. Furthermore, a higher rest heart rate was independently associated with arterial stiffness.
Abstract P281: Artificial Dendritic Cells Identify a Major Histocompatibility Complex Class 1 Subset that Activates T Cells in Hypertension
Authors: Chen Wei , Patrick David M , Barbaro Natalia R , Bernstein Kenneth E , Bernstein Kenneth E , Roeth Daniel , Kalkum Markus , Xiao Liang , Kirabo Annet , Harrison David G
Published: December 2018

We and others have shown an important role CD8+ cells in both experimental and human hypertension. CD8+ T cells are activated by antigens presented by major histocompatibility complex 1 (MHC1). C57BL/6 mice express two MHC1, referred to as H2-Kb and H2-Db. To identify antigenic peptides responsible for hypertension, we made two transgenic mice lacking the transmembrane domains of MHC1 and an added His-tag to the modified MHC1. These transgenes are driven by CD11c promoter, allowing expression in antigen presenting cells. The soluble Kb and Db (sKb and sDb) mice received 2 week infusions of sham or angiotensin II and their splenocytes placed in culture for two days. Ni-NTA beads were then used to bind the shed MHC-1 and these beads mixed with 10
6
T cells from other ang II infused mice (Figure panel A). CFSE dilution was used to monitor T cell proliferation. We found that sDb from ang II infused male mice, but not sham infused mice, potently stimulated proliferation of CD8+ T cells from ang II infused male mice. This degree of stimulation was significantly greater than that observed by sKb (Figure panel B). The shed MHC1 from female ang II-treated mice caused significantly less stimulation of female CD8+ T cells compared to male MHC1, in keeping with prior observations that female mice have less immune activation in hypertension. In summary, these data strongly suggest that unique antigens, presented in the context of H2-Db, are generated in hypertension. These data also suggest that there may be unique human lymphocyte antigens (HLAs), the analog of mouse MHC, that predispose to hypertension and related end-organ damage.
Abstract P290: Central Angiotensin II and Interleukin-1©¬ Upregulate Brain Tumor Necrosis Factor-¥å Converting Enzyme in the Rat
Authors: Cao Yiling , Yu Yang , Bell Balyssa B , Felder Robert B , Wei Shun-Guang
Published: December 2018

Tumor necrosis factor-α (TNF-α) converting enzyme (TACE/ADAM17) proteolytically cleaves the extracellular domain of transmembrane TNF-α to release the soluble form of TNF-α that contributes to augmented sympathetic nerve activity in heart failure (HF) and hypertension (HTN). Our previous study indicated that TACE is upregulated in the subfornical organ (SFO) and hypothalamic paraventricular nucleus (PVN), two key cardiovascular and autonomic brain centers, in rats with ischemia-induced HF, and is associated with TNF-α –induced sympathetic excitation in that setting. However, the mechanisms regulating TACE activity in the brain remain a mystery. The present study sought to determine whether angiotensin II (ANG II) or interleukin-1β (IL-1β), which increase in SFO and PVN in HF, upregulate TACE expression. Urethane anesthetized male SD rats underwent a 4- hour intracerebroventricular (ICV) infusion of artificial cerebrospinal fluid (aCSF), ANG II (100 ng/hr) or IL-1β (50 ng/hr). Some rats were euthanized to collect SFO and PVN tissues to measure TACE mRNA and protein; others were transcardially perfused with 4% paraformaldehyde to immunostain for TACE in SFO and PVN. ICV infusion of ANG II (n=6) or IL-1β (n=6) significantly (*p<0.05) increased TACE mRNA level (fold change) in SFO (2.54 ± 0.24* and 2.36 ± 0.21*, respectively) and PVN (2.39* ± 0.25 and 2.23 ± 0.20*, respectively) compared with ICV aCSF infusion. Western blot analysis also revealed a significant increase in the TACE protein (normalized to β-actin) in ANG II or IL-1β infused rats in SFO (0.31±0.06* and 0.26±0.05* respectively) and PVN (0.27 ± 0.04*, 0.24 ± 0.03*, respectively) compared with ICV aCSF-infused rats (SFO: 0.12±0.05; PVN: 0.09±0.04). Confocal images exhibited that TACE-like immunoreactivity was markedly higher in all four subdivisions of PVN and in SFO in rats treated with ICV ANG II or IL-1β, compared with aCSF-infused rats. These data indicate that ANG II and IL-1β upregulate TACE expression in SFO and PVN, suggesting that activation of the renin-angiotensin system and augmented inflammation may upregulate TACE expression in the SFO and PVN in HF and HTN. TACE-mediated ectodomain shedding in SFO and PVN may contribute to ANG II- and cytokine-driven sympathoexcitation in HF and HTN.
Abstract P303: Inhibition of Group IV Cytosolic Phospholipase A
2
α by Cytochrome P450 1B1-Estradiol Derived Metabolite 2-Methoxyestradiol Protects Against Angiotensin II-Induced Hypertension in Female Mice
Authors: Song Chi Young , Singh Purnima , Motiwala Mustafa , Shin Ji Soo , Bonventre Joseph V , Malik Kafait U
Published: December 2018

Previously we showed that the protection against angiotensin (Ang) II-induced hypertension and associated pathophysiological changes in female mice are mediated by cytochrome P450 (CYP) 1B1-estradiol (E2) generated metabolite 2-methoxyestradiol (2-ME). Also, we demonstrated that Ang II-induced hypertension is mediated by group IV cytosolic phospholipase A
2
α (cPLA
2
α) activation resulting in arachidonic acid (AA) release, and generation predominantly of eicosanoids with pro-hypertensive effects in male mice. This study was conducted to determine the contribution of cPLA
2
α/AA system, and its relationship to CYP1B1 in Ang II-induced hypertension in the female mice. Ang II infusion (700 ng/kg/min, s.c. micro-osmotic pump) for 14 days increased the systolic blood pressure (SBP), measured by tail-cuff in wild-type mice (cPLA
2
α
+/+
) but not in cPLA
2
α gene-disrupted mice (cPLA
2
α
-/-
) (151±8 vs. 126±9 mmHg,
P
<0.05, n=4-5). Ang II markedly increased SBP in ovariectomized (OVX) cPLA
2
α
+/+
mice but not in OVX cPLA
2
α
-/-
mice (175±3 vs. 121±2 mmHg,
P
<0.05, n=4-5). AA metabolism inhibitor 5,8,11,14-eicosatetraynoic acid (ETYA, 50 mg/kg, i.p. every 3
rd
day) attenuated (
P
<0.05) the Ang II-induced increase in SBP in intact (124±5 mmHg, n=5) and OVX cPLA
2
α
+/+
mice (124±5 mmHg, n=4). E2 attenuated Ang II-induced increase in SBP in OVX

Cyp1b1
+/+

mice, but not in OVX

Cyp1b1
-/-

mice (125±5 vs. 158±4 mmHg,
P
<0.05, n=3-4). Ang II-induced increase in SBP was also attenuated by ETYA in the

Cyp1b1
-/-

mice (129±7 vs. 185±7 mmHg,
P
<0.05, n=4). Ang II-induced increase in cPLA
2
α
+/+
activity examined in the kidneys was inhibited in

Cyp1b1
-/-

mice treated with 2-ME. Antagonists of prostaglandin (PG) E2 receptors EP1 (SC19220) and EP3 (L-798106) (28 μg/g, s.c. every 2
nd
day) minimized Ang II-induced increase in SBP in OVX cPLA
2
α
+/+
mice (126±2 and 127±5 vs. 175±3 mmHg, respectively, n=5). These data suggest that CYP1B1-E2 generated metabolite 2-ME protects against Ang II-induced hypertension by inhibiting cPLA
2
α activity and production of AA-derived PGE2 that exerts pro-hypertensive effects by stimulating EP1 and EP3 receptors. Thus, 2-ME and the drugs that selectively block EP1 and EP3 receptors could be useful for the treatment of hypertension and its pathogenesis in females.
Abstract P305: High Fat Diet With Normal Salt Feeding Does Not Change Renal m-TORC Signaling Activities In Male And Female Dahl Salt Sensitive Rats
Authors: Roxanne Fernandes , Galligan James J , Fink Gregory D , Xu Hui
Published: December 2018

We have previously reported that high fat diet (HFD) with normal salt feeding strongly promotes hypertension in both male and female Dahl salt sensitive (SS) rats. Only males, however, exhibit hypertension-associated renal inflammation/injury. The underlying mechanisms responsible for sex differences in HFD associated renal inflammation/injury are undetermined. It has been reported that increased activity of Mammalian Target of Rapamycin Complex 1/2 (mTORC) contribute to high salt diet associated hypertension and renal inflammation/injury in Dahl SS male rats. In this study, we determined whether HFD increases renal mTORC signaling in Dahl SS rats and if there are sex differences in signaling. Renal cortex and medulla were collected from rats fed a control diet (CD, 10% kcal fat) or a HFD (60% kcal fat) with 0.3% NaCl for 10, 17 or 24 weeks (WKs, starting at weaning, the time points correspond to pre-hypertension, developing hypertension and established hypertension phases of the model. Ratio (%) of expression of phospho-S6 ribosomal protein
Ser235/236
(pS6)/S6 (a marker for mTORC1), and phospho-AKT
Ser473
(pAKT)/AKT (a marker for mTORC2) were used to evaluate mTORC activity. The activity of mTORC1 in CD male cortex and female medulla were slightly increased at 17 and 24WKs of feeding compared to 10 WKs (22±4 and 16±7 vs 2±2, P<0.05), but HFD did not change the activity in cortex and medulla significantly in males (cortex,10±5, 14±5, 20±5; medulla 9±2, 18±6, 25±12) or females (cortex, 4±1, 12±3, 13±2; medulla 13±8, 22±8, 24±16) (P>0.5). The overall activity of mTORC2 in CD males and females were very low and mostly unchanged during 24WKs feeding, except for a slight increase in CD male cortex at 17WKs of feeding compared to 10WKs (4±1 vs 0.1±0.02, P<0.05). HFD caused only a slight increase of mTORC2 in medulla from males (4±1 vs 1±0.5) and a decrease in medulla from females (8±2 vs 0.9±0.3) (P<0.05) at 24WKs compared with 10WKs of feeding. Overall HFD did not cause marked changes in renal mTORC1/2 signaling activities. In addition, there were no significant differences in mTORC signaling activities between males and females. Therefore mTORC signaling pathways are not likely to contribute to the sex differences in HFD associated renal inflammation/injury in Dahl SS rats.