bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 081: Renal Sensory Nerves Increase Blood Pressure and Sympathetic Nerve Activity in 2-Kidney 1-Clip Hypertensive Mice
Authors: Ong Jason , Sved Alan F , Tan Roderick J , Rush Brittney M , Carattino Marcelo D , Stocker Sean D
Published: December 2018

Renal denervation lowers arterial blood pressure (ABP) in multiple clinical trials and some experimental models of hypertension. These antihypertensive effects have been attributed to the removal of renal afferent nerves. The purpose of the present study was to determine whether renal sensory nerves contributed to the 2-Kidney-1-Clip (2K1C) model of hypertension. 2K1C hypertension was produced in male C57Bl6 mice (12-14 weeks, Jackson Laboratories) by placement of a 0.5mm length of PTFE tubing (ID: 0.008” x OD: 0.014”) around the left renal artery. 2K1C mice (n=6) displayed an elevated ABP measured via telemetry (Day 0: 96±4mmHg vs Day 14: 115±3mmHg, P<0.05). Ganglionic blockade with hexamethonium (30mg/kg, ip) produced a greater fall in mean ABP at Day 14 vs Day 0 (Day 0: -38±4mmHg vs Day 14: -51±4mmHg, P<0.05). Ipsilateral vs contralateral kidneys of 2K1C mice had lower mass (0.072±0.01g vs 0.163±0.02g, respectively; P<0.05) and higher mRNA levels of several pro-inflammatory cytokines (IL1B, IL2, IL10, TNFa; P<0.05). Both total renal denervation (10% phenol) or selective denervation of renal afferent nerves (periaxonal application of 33mM capsaicin) at time of clipping resulted in a lower ABP than 2K1C mice at Day 14 (2K1C: 115±3mmHg, phenol: 104±2mmHg, capsaicin: 105±3mmHg; P<0.05). Direct recording of renal afferent nerve activity showed significantly greater discharge in 2K1C versus control mice (control: 2.2±1.3Hz vs 2K1C: 61±12Hz, n=3/group; P<0.05). Furthermore, electrical stimulation of renal afferent nerves in control mice produced a frequency-dependent increase in ABP (5Hz: 3±1mmHg, 10Hz: 7±1mmHg, 20Hz: 12±2mmHg, n=4/group; P<0.05). These responses were eliminated after ganglionic blockade with 5mg/kg chlorisondamine (5Hz: 1±1mmHg, 10Hz: 1±1mmHg, 20Hz: 1±1mmHg; P<0.05). Stimulus-triggered averaging of SNA during stimulation of renal afferent nerves (1 Hz, 200uA) revealed significant (P<0.05, n=4/group) increases in splanchnic (221±28%), renal (195+15%), and lumbar (234±32%) SNA. Interestingly, the latency to the peak SNA (142±10ms) suggests supraspinal pathways mediate the sympathoexcitatory response. These findings suggest 2K1C hypertension depends on renal sensory nerves and elevated SNA via supraspinal pathways.
Abstract 093: Salt Resistance and Renal Gpr83 Function
Authors: Zheng Xiaoxu , Konkalmatt Prasad , Asico Laureano , Jose Pedro A , Armando Ines
Published: December 2018

The G protein coupled receptor 83 (Gpr83) is an orphan receptor that has been associated with essential hypertension. Gpr83 is expressed in the kidney but its function is unknown. We found that Gpr83 is expressed in mouse renal proximal and distal convoluted tubules, as well as in human renal proximal tubule cells (hRPTCs). In C57Bl/6J mice on normal salt diet, the lack of one (Gpr83+/-) or both Gpr83 (Gpr83-/-) alleles resulted in an increase in systolic blood pressure (SBP, ~20 mm Hg (P<0.05; n=4/group, measured under anesthesia) compared with Gpr83+/+ littermates, suggesting that Gpr83 is needed to keep a normal BP. Renal specific Gpr83 silencing by the renal subcapsular infusion of Gpr83 siRNA (3 μg/day; 7 days) increased SBP in C57Bl/6J mice on a normal salt diet, relative to mice treated with nonsilencing siRNA (120±5 vs 98±6 mmHg; P<0.05; n=4/group). A high salt (4% NaCl) diet increased renal Gpr83 transcription by 2 fold (P<0.05; n=4/group) in SJL/J and BALB/c salt resistant mice, relative to C57Bl/6J salt sensitive mice.
Gpr83
gene down-regulation in mouse and human renal proximal tubule cells decreased about 50% the expression of nuclear factor of activated T cells 5 (NFAT5). The renal expressions of NFAT5 and the NFAT5 targets UT-A3, Tau T and BGT1 were increased by a high salt diet in salt resistant mice. The neuropeptide PEN (PEN), a ligand of Gpr83, is also expressed in hRPTCs (0.43±0.03 mRNA PEN/mRNA GAPDH). We found that Gpr83 stimulation with PEN (1 μM, 3 hr) decreased the transcription of NFAT5 (0.59±0.05 vs 1.0±0.2fold, P<0.05; n=3-4/group), which was abolished by Gpr83 silencing. Our results suggest that Gpr83 may protect against the development of salt sensitivity. Gpr83 mediated regulation of NFAT5 may play a role in salt-sensitive hypertension.
Abstract 108: Sympathetic Inhibition Prevents the Homing of Specific Memory T Cells to the Bone Marrow and the Development of Repeated Hypertension
Authors: Xiao Liang , Carmo Luciana S , Itani Hana A , Foss Jason D , Chen Wei , Harrison David G
Published: December 2018

Effector memory T cells (T
EM
cells) play a crucial role in hypertension. Formed during an initial blood pressure surge, T
EM
cells can reside in bone marrow (BM) in a quiescent state for prolonged periods. Upon re-exposure to a hypertensive stimulus, these cells can be reactivated and aggravate hypertension and renal damage. Hypertension is also associated with elevated sympathetic outflow. We hypothesized that sympathetic nerves promote accumulation and reactivation of hypertension-specific T
EM
cells in the BM. We performed unilateral superior cervical ganglionectomy (SCGx) in C57BL/6 mice, causing sympathectomy of the forelimb on the surgical side. After 2-week Ang II infusion (490 ng/kg/min s.c.), 5х10
6
BM cells from both the SCGx and control limbs were isolated and co-cultured with 0.5х10
6
dendritic cells from other hypertensive mice. We found 30% fewer CD8
+
T cell proliferation in the SCGx BM than intact side (1.9±0.2 vs. 2.8±0.1х10
4
, p<0.01), but no difference in CD4
+
T cells. To study mechanisms involved in T cell homing, 10
7
pan T cells were isolated from CD45.2 mice after Ang II infusion and adoptively transferred to CD45.1 mice with unilateral SCGx. Flow cytometry indicated that 35% fewer donor CD8
+
T
EM
cells homed to the SCGx than the intact BM 7 days after transfer (5.3±0.8 vs. 8.1±1.5 per 10
4
total T cells, p<0.05). We further determined if systemic sympatho-inhibition during T cell homing is protective against future hypertensive stimuli. We blocked systemic sympathetic outflow by injecting an inhibitory designer receptor exclusively activated by a designer drug (Gi-DREADD) into the rostroventral lateral medulla (RVLM) and performed pan-T cell adoptive transfer 10 days later. The DREADD ligand CNO was given in drinking water 3 days before till 7 days after adoptive transfer. After washout, mice received low dose Ang II infusion (140 ng/kg/min) for 2 weeks. As measured by radiotelemetry, mice with control injection developed hypertension (160±7 mmHg). However, mice with Gi-DREADD expressed in RVLM remained normotensive (136±3 mmHg, p<0.05 vs. control). These data define a novel role of sympathetic nerves in regulation of memory T cell trafficking, and sympathetic inhibition may have long term protective effect beyond lowering blood pressure.
Abstract 116: Epigenetic Regulation of Human Angiotensinogen (hAGT) Gene by SNPs.
Authors: Perla Sravan , Mopidevi Brahmaraju , Sivankutty Indu , Jain Sudhir , Jain Kavita , Kumar Ashok
Published: December 2018

Hypertension is a complex disease caused by a combination of genetic and environmental factors. Renin angiotensin system (RAS) dysfunction leads to essential hypertension. Polymorphisms in the AGT gene increase RAS activity and cause blood pressure elevation. We and others have shown that SNPs in the promoter of hAGT gene can be arranged mainly in two haplotypes. Variants -1670A, -1562C, and -1561T always occur with -6A and form the haplotype-l (Hap-l), while variants -1670G, -1562G, and -1561G always occur with -6G and constitute haplotype-ll (Hap-ll). We have made transgenic animals containing these two haplotype blocks and shown that Haplotype-I is the risk haplotype and increases hAGT expression in TG animals. We hypothesized that these SNPs, when present together as Hap-I or Hap-II in transgenic mice (TG) may lead to haplotype dependent DNA methylation of CpG sites in the promoter of the hAGT gene. Methylation patterns may modulate gene transcription at an epigenetic level and this, in turn, could be dependent on individual polymorphisms. Hypo-methylation is associated with higher gene expression and hyper methylation is associated with lower gene expression. We have found that the number of methylated CpG sites in the hAGT promoter is decreased after high salt (HS) treatment in the liver of Hap-I TG animals as compared to the liver of Hap-I animals. In the liver of Hap-I animals, the hAGT promoter CpG sites are methylated at -460,-434,-386,-261,+42, whereas after high salt treatment, the promoter DNA methylation is observed at -460,-386,+65. On the other hand methylation of hAGT promoter CpG sites in the liver of Hap-II are increased as compared to the liver of Hap-II after high salt treatment. In the liver of Hap-II animals, the CpG sites are methylated at -386,-346,+65, whereas after high salt treatment, methylate sites are observed at -386,-261. Results of this experiment show that methylation of CpG sites is reduced in the DNA obtained from high salt treated liver of both haplotypes. However, DNA from high salt treated liver of Hap-II animals had least number of methylated CpG sites and therefore open chromatin that leads to higher expression of the hAGT gene in this haplotype.
Abstract 117: Inhibition of DNA Methyltransferase Attenuates Salt-Sensitive Hypertension & Renal Disease in Dahl Salt Sensitive Rats
Authors: Dasinger John Henry H , Pan Xiaoqing , Abais-Battad Justine M , Alsheikh Ammar J , Lund Hayley , Fehrenbach Daniel J , Liu Pengyuan , Liang Mingyu , Mattson David L
Published: December 2018

DNA methylation plays a key role in the regulation of gene expression. In the present study, we investigated the impact of changes in DNA methylation in Dahl Salt Sensitive (SS) rats fed a casein-based AIN-76A chow containing low salt (LS) or high salt (HS). In addition to the hypertension and renal damage which occurs following 14 days of HS feeding, we assessed 5-Methylcytosine levels at single-base resolution by reduced representation bisulfite genome sequencing (RRBS) of DNA from the renal outer medulla of SS rats. Several hundred differentially methylated regions (DMRs) were identified between the SS rats on the LS or HS diet. Approximately a quarter of the DMRs were located in transcriptional start site regions, a third in intragenic regions, and the remainder in intergenic regions. To investigate the functional role of methylation in SS hypertension, we administered decitabine (DNA methyltransferase inhibitor, 5mg/kg, ip) or vehicle at Days 0, 3, 5, and 7 during a 14 day HS period. While there was no difference in MAP during the LS period (127±3 vs. 126±6 mmHg, vehicle vs. decitabine, n=9), at the end of 2 weeks of HS there was a significant increase in MAP in the vehicle SS rats compared to the decitabine-treated rats (163±6 vs 141±4 mmHg,
p<0.0001
). This protective effect in MAP in the decitabine-treated group also exhibit a reduction in albumin excretion rate compared to controls (119±19 vs. 182±24 mg/day,
p<0.01
). Upon analysis of the immune cell infiltration profile, there was a reduction in number of CD45+ total leukocytes, CD11b/c+ monocytes/ macrophages, and CD45R+ B cells in the decitabine-treated animals compared to controls. Decitabine administration also significantly reduced renal histological damage as indicated by the presence of outer medullary protein casts compared to vehicle-treated rats (6.8±1.0% vs. 10.6±1.3%,
p
<0.05). Lastly, methylation rate was significantly reduced in decitabine-treated rats by 40% compared to the control levels. In summary, the inhibition of methylation by decitabine attenuated MAP and markers of renal damage in response to a HS diet suggesting that DNA methylation could be a potential therapeutic target for the treatment of hypertension and renal damage.
Abstract 134: The Effect of Low Birth Weight on the Microcirculation in the 1
St
Year of Life
Authors: Goloba Muti E , Raghuraman Rajendra , Khan Uzma , Brown Amelia , Klein Monique , Botros Nansi , Duffy Donovan , Wang Duolao , Manyonda Isaac , Antonios Tarek F
Published: December 2018

Introduction:
Capillary rarefaction, defined as a reduction in capillary density, is an established hallmark of essential hypertension. Low birth weight (LBW) infants, known to have an increased risk of developing hypertension as adults, were unexpectedly found to have a significantly higher capillary density at birth when compared to normal birth weight (NBW) infants. We therefore hypothesised that there is a microcirculatory window in the 1
st
year of life of LBW infants, during which a process of extensive capillary loss, known as “hyperpruning”, occurs.

Methods:
The George’s Capillary Rarefaction Offspring Study (G-CROS) is a longitudinal, multi-centre study of which 284 infants were NBW, born at term, and 77 were LBW. Intravital microscopy was used to measure the functional (also known as basal) capillary density (BCD), and the structural (also known as maximal) capillary density (MCD) at birth, 3 months, 6 months and 12 months.

Results:
LBW infants had a significantly higher capillary density at birth when compared to NBW infants (
p
< 0.0001). NBW infants showed a gradual reduction in capillary density between birth and 12 months, with their greatest reduction occurring between birth and 3 months (BCD mean difference
=
-27.62 cap/field, p <0.0001 and MCD mean difference = -31.49 cap/field, p <0.0001). Similarly, the most significant reduction in BCD and MCD, in the LBW cohort, was between birth and 3 months (BCD mean difference
=
-47.01 cap/field,
p
< 0.0001 and MCD mean difference
=
-48.01 cap/field,
p
< 0.0001). However, within this same 3-month interval, LBW infants demonstrated a significantly higher percentage reduction in BCD (mean difference = 7.81%,
p
= 0.0194) and MCD (mean difference = 8.29%,
p
= 0.0361) when compared to NBW infants.

Conclusions:
Once again, LBW infants were shown to have a higher capillary density than NBW infants at birth. However, this study has shown for the first time that there appears to be a microcirculatory window in the first 3 months of life during which LBW infants undergo capillary “hyperpruning”. Further follow-up studies are required to investigate the role of early capillary rarefaction in the pathogenesis of hypertension, as well as the mechanisms orchestrating these early microcirculatory changes.
Abstract 138: White AdiposeTissue in the Preeclamptic-Like BPH/5 Mouse Has a Unique Inflammatory Molecular Signature During Early Pregnancy
Authors: Sones Jennifer L , Reijnders Dorien , Olson Kelsey , Redman Leanne
Published: December 2018

Introduction:
Preeclampsia (PE) is a hypertensive disease of pregnancy that affects an estimated 8% of women worldwide. The underlying mechanism is unknown; however, maternal obesity increases the risk of PE by 30% and may contribute to increased inflammation seen in preeclamptic mothers.

Hypothesis:
To better understand the role of white adipose tissue (WAT) signaling on adverse pregnancy outcomes, we used the obese PE-like BPH/5 mouse to test the hypothesis that visceral WAT has an inflammatory phenotype prior to the presentation of the maternal hypertensive syndrome

Methods:
Visceral WAT adjacent to the reproductive tract (“reproductive WAT”) was harvested from ad libitum fed control C57 and BPH/5 female mice (8 weeks of age) at embryonic day (e) 7.5. RNA was isolated and subjected to Next Generation Sequencing (NGS) using an Illumina platform. DAVID pathway analysis was performed to annotate differentially expressed genes. BPH/5 female mice were calorie restricted by 25% from e0.5 until e7.5 and reproductive WAT was collected for RNA isolation and quantitative real-time PCR. The calorie restriction paradigm has been shown to significantly reduce reproductive WAT accumulation throughout gestation in BPH/5 mice.

Results:
NGS of e7.5 reproductive WAT revealed 899 genes that were significantly dysregulated in BPH/5 vs C57. DAVID showed upregulation of lipid metabolic processes in both C57 and BPH/5 pregnant reproductive WAT, while 18 inflammation-related pathways were significantly upregulated in reproductive WAT of pregnant BPH/5 mice compared to pregnant C57 (FDR<=5%, adjP<=0.1, FC >=1.5). Quantitative real-time PCR confirmed upregulation of PE- related inflammatory genes, interleukin (IL)-6, IL-15 and Ptgs2, in e7.5 BPH/5 reproductive WAT vs C57 (n=6, p<0.05). Seven days 25% calorie restriction during pregnancy significantly reduced this elevated IL-6 and Ptgs2 mRNA expression in reproductive WAT (n=6, p<0.05)

Conclusions:
These data show that obese PE-like BPH/5 mice have elevated inflammatory gene expression in reproductive WAT before placenta formation. Further investigations are needed to confirm if the reduction in IL-6 and Ptgs2 in reproductive WAT by calorie restriction improves fetoplacental and maternal outcomes in BPH/5 mice
Abstract P107: Real-World Azilsartan efficacy Assessment Study in Indian Hypertensive patients (RAAS-India)
Authors: Jadhav Uday , DALAL JAMSHED
Published: December 2018

Background:
Azilsartan, a potent angiotensin type 1 receptor blocker (ARB) has limited data on the efficacy and safety in hypertensive subjects in India.

Methods:
Retrospective audit based on data from the prescribing physicians on efficacy of Azilsartan monotherapy or in combination with other antihypertensive drugs. Office blood pressure (BP) measurement prior to initiation of Azilsartan, at 1
st
and 2nd follow-up visits were recorded. Two-tailed paired
t
-test and nonparametric Wilcoxon signed-rank test were used for statistical analysis.

Results:
Data of 1397 patients was collected of which 844 patient were eligible for analysis at baseline. Mean age was 54.7 years. Azilsartan was started as monotherapy in 64.6% patients and as an add-on to antihypertensive therapy in 35.4% patients. Significant reduction of 19.6 mm Hg in mean Systolic BP (baseline: 162.6
+
15.6 mm Hg to visit 1: 143
+
13.7 mm Hg, p<0.001) and 30.2mm Hg in mean Systolic BP (baseline: 162.6
+
15.6 mm Hg to visit 2: 132.4
+
10.7, p<0.001) was observed. Significant reduction of 9.2 mmHg in mean diastolic BP (baseline: 95.4
+
10 mmHg to visit 1 at mean day 36 : 86.2
+
7.1 p<0.001) and 13.5mmHg in mean diastolic BP (baseline: 95.4
+
10 mmHg to visit 2 at mean day 71: 81.9
+
5.7 p<0.001) was observed. Azilsartan in the prescribed dose was well tolerated. Most common adverse event reported were Nausea 10 (1.2 %), metallic taste 10 (1.2 %) and giddiness 8 (0.9 %). No serious adverse events were reported.

Conclusion:
Azilsartan alone or with other antihypertensive drugs significantly reduced BP in Indian hypertensive patients in a real world setting. Azilsartan was well tolerated without any major or serious adverse events.
Abstract P121: The Effect of Intensive Blood Pressure Targets on Mortality in Chronic Kidney Disease Patients: An Individual Patient Data Meta-Analysis of Randomized Controlled Trials
Authors: Aggarwal Rahul , Chiu Nicholas , Petrie Benjamin , Mirzan Haares , Steinkamp Jackson
Published: December 2018

Introduction:
Current evidence is conflicting on whether intensive blood pressure (BP) management, specifically with targets of <130/90 mmHg, produces a mortality benefit in non-diabetic chronic kidney disease (CKD) patients. MDRD, AASK, and SPRINT are the largest randomized controlled trials available investigating intensive targets in CKD patients. The MDRD and AASK recruited exclusively CKD patients, while SPRINT had a large CKD subpopulation within its cohort. MDRD assigned patients to a standard mean arterial pressure (MAP) target of 107 mmHg vs. an intensive target of 92 mmHg (achieved BPs of 134/81 mmHg vs 126/77 mmHg, respectively). AASK assigned African American patients to a MAP of 102-107 mmHg vs. <92 mmHg (achieved BPs of 141/85 mmHg vs. 128/78 mmHg, respectively). SPRINT compared a systolic target of 140 mmHg vs. 120 mmHg (achieved BPs of 137/74 vs. 123/67 in the CKD subpopulation). Our study pools individual patient data (IPD) from these three trials.

Methods:
IPD for all patients in MDRD, AASK, and SPRINT were obtained and screened for non-diabetic CKD patients. The resulting 4537 patients were assigned to their original randomized intervention group – 2258 patients to the standard group and 2279 to the intensive group. The primary outcome was mortality. Statistical analysis for primary outcome was performed with Cox proportional hazards models stratified by clinical site/center. Interactions between outcome and trial enrollment were also assessed.

Results:
There was no statistically significant difference in mortality rate between intensive and standard BP control (HR: 0.84; 95% CI: 0.66, 1.07; p = 0.148). 126 patients (5.5%) in the intensive group and 149 patients (6.6%) in the standard group expired. No interaction was present between treatment and trial enrollment (MDRD, AASK, SPRINT) (p-value of interaction: 0.080).

Conclusion:
Intensive BP targets do not improve mortality for non-diabetic patients with CKD. Further investigation needs to be performed to determine if this lack of benefit is consistent across subpopulations.
Abstract P122: Early Renal Allograft Function and Hypertension after Kidney Transplantation
Authors: Tantisattamo Ekamol , Dejhansathit Siroj , Vutthikraivit Possawat , Mopuru Haritha , Ratanasrimetha Praveen , Rattanavich Rungwasee , Pullalarevu Raghavesh
Published: December 2018

Background:
Poor renal allograft function is associated with post-transplant hypertension (HTN). The association between allograft function at an early post-transplant period and HTN is unclear.

Method:
Of 70 transplant recipients, the difference between serum creatinine (SCr) at the discharge date from kidney transplantation (KT) and those at pre-transplantation (ΔCr) divides the study population into Group 1 (ΔCr ≥80%) and Group 2 (ΔCr <80%). Cox proportional hazard regression is performed to find the association between different ΔCr and post-transplant systolic HTN (SHTN) and diastolic HTN (DHTN) defined by SBP ≥130 and DBP ≥80 mmHg, respectively during a 96-week follow-up period.

Results:
Mean age±SEM is 52.66±1.43 years and 58.6% is male. Mean length of stay is 8.50±1.35 days. Thirty patients (43%) are in the Groups 1. Mean absolute ΔCr is greater in the Group 1 (ΔCr -7.41±0.43 vs. -3.49±0.42, p <0.001). Compared to Group 1, Group 2 has a 17.1% increased risk of SHTN (HR 1.171, 95%CI 0.715 to 1.918, p 0.530, Fig1A), but has a 24.5% decreased risk of DHTN (HR 0.755, 95%CI 0.417 to 1.367, p 0.353, Fig1B). After adjusting for age, gender, type of induction immunosuppressants, type of KT, and BMI, Group 2 has an 80.2% greater risk for SHTN (HR 1.802, 95%CI 0.976 to 3.325, p 0.060, Fig2A), but has a 6.8% lowered risk of DHTN (HR 0.932, 95%CI 0.467 to 1.857, p 0.840, Fig2B).

Conclusions:
The degree of change in SCr during immediate post-KT does not predict post-transplant SHTN and DHTN. However, patients with better renal allograft function appear to become less SHTN but more DHTN. The pattern of association between long-term allograft function and SHTN and DHTN needs further studies.
Abstract P133: Obstructive Sleep Apnea May Exacerbate Carotid Artery Stiffness in Individuals With Chronic Obstructive Pulmonary Disease
Authors: Luehrs Rachel E , Moreau Kerrie L , Aloia Mark , Weinberger Howard D , Make Barry , Bowler Russell , Crapo James D , Wamboldt Frederick , Hoth Karin F , Pierce Gary L
Published: December 2018

Background:
Chronic obstructive pulmonary disease (COPD) and obstructive sleep apnea (OSA) are two of the most prevalent respiratory disorders in adults and are both independently associated with increased cardiovascular disease (CVD) risk. The coexistence of OSA in individuals with COPD is termed “overlap syndrome” and occurs in up to 65% of all COPD patients. Individuals with overlap syndrome have more frequent COPD exacerbations, poorer quality of life and increased CVD morbidity than patients with either COPD or OSA alone. COPD and OSA are both independently associated with increased carotid artery stiffness, a predictor of CVD events. However, the interaction of COPD and OSA on carotid artery stiffness remains unknown. We hypothesized that a) carotid artery stiffness would be greater in patients with overlap syndrome than COPD patients without OSA and non-COPD controls and b) overlap syndrome patients that reported using continuous positive airway pressure (CPAP) would have lower carotid artery stiffness than overlap patients not using a CPAP.

Methods:
Ninety-nine former smokers (n=62 with COPD and n=37 non-COPD controls; age 70 ± 7 yrs; 60M/39F) completed measurements of pulmonary function and common carotid artery stiffness and were retrospectively stratified by self-report of previous clinical diagnosis with OSA.

Results:
Participants with overlap syndrome had significantly greater carotid β-stiffness than COPD patients without OSA, non-COPD controls with OSA and non-COPD controls without OSA (overlap 5.7 ± 0.71 vs COPD only 3.9 ± 0.20, OSA only 3.6 ± 0.31, controls 3.7 ± 0.23 U; F=6.245, p<0.01) and this difference remained significant after adjustment for body mass index (p=0.01). There were no differences in blood pressure, heart rate, resting oxygen saturation (SpO
2
) or nocturnal mean SpO
2
in those with overlap syndrome vs. the other groups. Furthermore, there was no difference in carotid β-stiffness between overlap syndrome patients that reported using vs. not using a CPAP (5.98 ± 0.7 vs 5.53 ± 1.9 U, p=0.80).

Discussion:
These data suggest that OSA exacerbates carotid artery stiffening in COPD patients and that carotid artery stiffness may be one mechanism that mediates the increased CVD risk observed in COPD patients with overlap syndrome.
Abstract P136: A Case Report of Aortic Dissection With Higher Level of Pulse Wave Velocity
Authors: Liu Jinbo , Li Tianrun , Liu Huan , Zhao Hongwei , Huang Wei , Wang Lichun , Song Yuejie , Yu Xiaolan , Wang Hongyu
Published: December 2018

Background:
Hypertension was an important risk factor of aortic dissection. Arteriosclerosis evaluated by arterial stiffness was the basic patho-physiological change of hypertension. We reported a case of aortic dissection with higher level of arterial stiffness.

Methods:
A patient (women, 62-year old) visited Peking University Shougang Hospital with sudden intense back pain for 3 hours on November 17, 2017. Her medical history was hypertension with treatment of levamlodipine 2.5mg/day, without regular monitoring.

Results:
Computed tomography angiography (CTA) indicated a cute Stanford B Type thoracic aortic dissection. The results of white blood cell count, platelet count, electrolytes, urea, creatine, erythrocyte sedimentation rate and liver function tests were all normal with higher level of D-Dimer of 13.5mg/L (reference range <1.0mg/L). Cardiac ultrasound showed no abnormal signs of aortic valve reflux and ventricular wall activity, with normal ejection fraction (59%). Electrocardiogram showed signs of cardiac ischemia with changes of T waves. Thoracic endovascular aortic repair therapy (TEVAR) was performed on November 18, 2017 and the back pain was relaxed. However, the patient had intermittent back pain on November 21, 2017 with recurrent aorta dissection showed by CTA. TEVAR was performed again immediately on November 21, 2017. Finally, her treatment was nifedipin controlled-release 30mg/day, irbesartan and hydrochlorothiazide 150mg/12.5mg/day, bisoprolol fumarate 3.75mg/day, terazosin hydrochloride 2mg/day, aspirin 100mg/day, with systolic blood pressure 90-100mmHg and heart rate of 55-60 beats per minute. Arterial stiffness showed that left brachial ankle pulse wave velocity (ba-PWV) of the patient was 2138cm/s (reference range <1400cm/s) and the right ba-PWV was 2018cm/s (reference range <1400cm/s).

Conclusions:
Our present case report showed that hypertension subject with higher level of pulse wave velocity was more likely to happen aortic dissection, so arterial stiffness should be evaluated during the management of hypertension. If we could find the abnormality of arteriosclerosis earlier, we could carry out some management to reduce arterial stiffness, and aortic dissection might be prevented in future.
Abstract P140: Leptin Supplementation Restores Endothelial Function and Vascular Adrenergic Tone via Direct Endothelial Mechanisms and Sympato-Activation, Respectively, in a Mouse Model of Congenital Lipodystrophy
Authors: Bruder-Nascimento Thiago B , Faulkner Jessica , Kennard Simone , Chen Weiqin , Belin de Chantemele Eric
Published: December 2018

Absence of adipose tissue in congenital generalized lipodystrophy (CGL) leads to metabolic and cardiovascular disease and reduced leptin levels. We tested the hypothesis that leptin supplementation restores endothelial function and reduces vascular adrenergic response respectively via acting directly on endothelial cells (EC) and desensitizing vascular adrenergic response, in CGL mice. Deletion of Berardinelli-Seip gene (Bscl2) was used to mimic the human pathology in mice. Bscl2-/- mice showed reduced adipose mass [% of fat: wild-type (WT): 8.6 ± 0.3 vs Bscl2-/-: 2.4 ± 0.1, P<0.05], leptin levels [ng/mL: WT: 4.0 ± 0.3 vs Bscl2-/-: 0.3 ± 0.1, *P<0.05], impaired aortic endothelium-dependent relaxation to acetylcholine (Ach): [% relaxation: WT: 71.9 ± 1 vs Bscl2-/-: 49.9 ± 2*, *P<0.05], increased vascular contractility to phenylephrine (Phe), associated with upregulation of α-adrenergic receptors, high mean arterial pressure (MAP) and reduced heart rate (HR): [(mmHg: WT: 100.7 ± 5 vs Bscl2-/-: 109.5 ± 6*), (bpm: WT: 592.7 ± 9 vs Bscl2-/-: 542.9 ± 15*) *P<0.05]. Moreover, Bscl2-/- mice shown increase in Nox1 gene expression and increases in vascular
-
O
2
and H
2
O
2
levels. Both chronic (in vivo) and acute (organ bath) leptin supplementation or NOX 1 inhibition (GKT771, 10μM) restored endothelial function in addition to reducing endothelial Nox1 expression and ROS production, which suggest a direct role for leptin in endothelial cells. EC leptin receptor deficiency induced an endothelial dysfunction in mice, which was restored by Nox1 inhibition, further supporting a direct role of leptin in endothelial cells. In parallel, leptin supplementation reduced vascular adrenergic tone and aortic α-adrenergic receptors and increased in HR without changing MAP. Our data show that leptin replacement restores endothelial function and reduces vascular adrenergic tone in mouse model of congenital lipodystrophy by acting directly in EC and desensitizing vascular adrenergic response, respectively. This highlight the importance of leptin in the maintenance of vascular homeostasis and presents leptin as a potential therapeutic avenue for the treatment of vascular diseases associated with low leptin levels.
Abstract P144: Characterization of Vascular Endothelial and Smooth Muscle Exosomes and Their Role in Vascular SignalTransduction
Authors: Boyer Michael , Kimura Yayoi , Eguchi Satoru , Rizzo Victor
Published: December 2018

Vascular homeostasis is predicated on the integral interaction between a myriad of cell types. Principle to homeostasis is the interaction between the endothelial cells (ECs) of the intima and the vascular smooth muscle cells (VSMCs) of the media. Disruption of the tightly controlled communication between these cell types can lead to development of pathophysiology. In the past decade, much study has been given to a novel means of intercellular communication in exosomes. Exosomes are nanovesicles derived from endosomal compartments capable of carrying functional proteins, small RNAs and lipids into extracellular space. Initially characterized in the field of immunology, little is known on exosomes within vascular biology. Therefore, we present an
in vitro
characterization of rat EC and VSMC exosomes and evidence of EC exosomes as initiators of signal transduction within VSMCs. Primary rat aortic EC and VSMC exosomes isolated in serum-free conditions displayed characteristic size, shape and protein content of exosomes as analyzed through nanoparticle tracking analysis, electron microscopy, and western blot. Interestingly, alpha smooth muscle actin was found to be cell-specific for VSMC exosomes. Proteomics identified differential expression between EC and VSMC exosomes suggesting altered cargo between the two populations. Incubation of EC exosomes with VSMC identified a dose- and time-dependent increase in the expression of vascular cell adhesion molecule 1 in VSMC with a maximum at 10
9
EC exosomes and 6 hours through immunoblot analysis. In addition, EC exosomes stimulated expression of early growth response protein 1 at 1-3 hours, suggesting activation of pro-inflammatory gene expression. EC exosomes stimulated a maximal rate of protein synthesis at 3 hours in VSMC (3.53 fold increase over control, p=0.016) determined through puromycin incorporation showing qualitative changes in cell function. Analysis of cell volume identified a hypertrophic response in VSMC after exosome incubation as well, displaying a role for exosomes in vascular hypertrophy. In conclusion, we present a broad characterization of exosomes in vascular biology and a functional crosstalk mechanism between RAEC and VSMC through exosome transfer.
Abstract P152: Exercise Training Upregulates Nrf2 Gene Expression in the RVLM of Mice With Heart failure
Authors: Wafi Ahmed M , Gao Lie , Yu Li , Zucker Irving H
Published: December 2018

Sympatho-excitation is an important hallmark in the heart failure (HF) state. It is known that HF is associated with global oxidative stress, which contributes to pathogenesis of sympatho-excitation. Central reactive oxygen species (ROS) accumulate within neurons and evoke alterations in protein function, leading to an enhanced neuronal excitability, thereby evoking the higher level of sympathetic activation in HF. On the other hand, exercise training (ExT) is associated with a reduction of oxidative stress, in part by upregulation of antioxidant enzymes. However, the link between ExT and antioxidants in the brain of animals with HF has not been explored. In this study, we hypothesized ExT enhances transcriptional activation of Nuclear factor (erythroid-derived 2)-like 2 (Nrf2), a master transcription factor that modulates a wide array of antioxidant enzyme gene expression, in the rostral ventrolateral medulla (RVLM). Sham and coronary artery ligation-induced HF mice were divided into either sedentary (Sed) or ExT groups. After 6 weeks of treadmill training, maximal exercise tolerance, urinary norepinephrine (NE) and Nrf2 as well as NAD(P)H dehydrogenase [quinone] 1 (NQO1) gene expression were evaluated by real time RT-PCR. We found that: (1) HF mice displayed significantly lower maximal distance than Sham mice, which was improved by ExT (maximal distance: Sham- Sed 255 ± 34 m (n = 6), HF Sed 148 ± 37 (n = 5), HF ExT 304 ± 34 (n = 6), p<0.05 HF Sed vs HF ExT); (2) urinary NE concentration was significantly higher in HF Sed group which was attenuated by ExT (urinary NE: Sham Sed 442 ± 38 (n = 6), HF Sed 596 ± 41 (n = 5), HF ExT 465 ± 38 ng/ml (n = 6), p<0.05); (3) ExT attenuated HF- induced lower Nrf2 and NQO1 gene expression and also upregulated gene expression of these two proteins in Sham mice (Nrf2: Sham Sed 0.92 ± 0.10 (n = 6), Sham ExT 1.32 ± 0.10 (n = 6), HF Sed 0.60 ± 0.11 (n = 5), HF ExT 0.95 (n = 6), p<0.05), (NQO1: Sham Sed 0.98 ± 0.12 (n = 6), sham ExT 1.39 ± 0.12 (n = 6), HF Sed 0.60 ± 0.13 (n = 5), HF ExT 1.07 ± 0.12 (n = 6), p < 0.05). These data suggest that upregulation of Nrf2 gene expression could be a mechanism by which ExT exerts its beneficial effects on sympathetic tone in HF. Further studies are required to investigate how ExT-induced Nrf2 upregulation alters sympathetic nerve activity in HF.
Abstract P157: Immunophenotyping Human Hypertension Using Single Cell Mass Cytometry
Authors: Alexander Matthew R , Dale Bethany L , Wogsland Cara E , Elijovich Fernando , Irish Jonathan M , Madhur Meena S
Published: December 2018

Hypertension is the leading risk factor for global morbidity and mortality. Emerging evidence in animal models implicates a variety of innate and adaptive immune cells in hypertension pathogenesis, and limited studies in humans have demonstrated increased circulating memory and senescent T cells in hypertensive individuals. However, the abundance and role of specific immune cells in human hypertension remains unclear. To perform unbiased high dimensional profiling of peripheral immune cells in humans, we developed and validated a novel approach using mass cytometry to detect 31 extracellular markers at the single cell level. Initial results in a normotensive individual using this panel revealed robust identification of major circulating immune cell subsets including CD4
+
cells (60% of CD45
+
cells), CD8
+
cells (21%), γδ T cells (6%), CD25
+
CD127
lo
T regulatory cells (5%), CD19
+
B cells (3%), and CD11c
+
myeloid cells (3%). Unsupervised viSNE plots using this panel of 31 markers demonstrates grouping of distinct circulating immune cell subsets based on similarity of marker levels
(Figure)
. We are extending this analysis to peripheral blood mononuclear cells collected from 16 additional control and 17 hypertensive individuals matched for age, gender, race, and BMI to determine differences in the abundance of known and potentially novel circulating immune cell subsets. The combination of high dimensional mapping and unsupervised computational analysis will permit rigorous immunophenotyping of human hypertension and potentially identify novel therapeutic targets.
Abstract P159: Neural Inflammation Results in Cardiac Sympathetic Afferent Sensitization in Post-Myocardial Infarction Rats
Authors: Hong Juan
Published: December 2018

Sympatho-excitation is a hallmark of the chronic heart failure (CHF) state. The enhanced cardiac sympathetic afferent reflex (CSAR) contributes to the elevated sympathetic tone in CHF. However, the mechanisms underlying the CSAR sensitization have not been fully discovered. We hypothesized that myocardial infarction (MI) will trigger a neuro-inflammatory cascade that includes macrophage activation in thoracic dorsal root ganglia (DRG), which results in cardiac spinal afferent sensitization. Our immunofluorescence data (
Figure 1
) demonstrated that, the number of Iba-1-positive (a macrophage marker) cells in T1-T4 DRGs of MI rats was significantly increased at 1 week and lasted at least 8 weeks post MI. RNA-seq analysis data in T1-T4 DRGs of MI rats also showed upregulated mRNA expressions of several macrophage-activation related genes such as IRF7, RGD1309362, slfn4 and lfit1. In the in vitro experiments, after the 50B11 DRG cells were co-cultured with LPS-pretreated BV2 cells (activated microglia/macrophage), the protein expressions of voltage-gated potassium channel isoforms (Kv1.4, Kv4.2, Kv4.3 and Kv3.4) were largely reduced, which explains how macrophage activation cause DRG neuronal sensitization. Lastly, our data showed that 4-week treatment with minocycline (a macrophage inhibitor, 40 mg/kg/day) via drinking water markedly reduced the enhanced CSAR in the post-MI rats (MAP, 30.2±2.6 vs. 18.0±1.9 mmHg, n=6, p<0.01; HR, 43.5±5.5 vs. 12.5±2.0 bpm, n=6, p<0.01; RSNA, 169.2±19.5 vs. 77.8±4.6% baseline, n=6, p<0.01). These data suggested that macrophage infiltration in sensory ganglia contributes to the sensitization of the CSAR in the post-MI state.
Abstract P171: Occupational, Sport and Leisure Physical Activity Have Contrasting Effects on Neural Baroreflex Sensitivity. The Paris Prospective Study III
Authors: Climie Rachel , Boutouyrie Pierre , Chaussade Edward , Perier Marie-Cecile , Plichart Mathew , Offrendo Lucile , Guibout Catherine , van Sloten Thomas , Thomas Frederique , Pannier Bruno , Sharman James , Laurent Stephane , Jouven Xavier , Empana Jean-Philippe
Published: December 2018

Background:
Physical activity (PA) is beneficial for baroreflex sensitivity (BRS), but it is unclear whether the type of PA has similar effects on the neural (nBRS) or vascular (carotid stiffness) components of BRS. We sought to determine this in healthy adults from a community- based study via assessment of occupational (OPA), sport (SPA), leisure (LPA) and total PA (TPA).

Methods:
In 8649 adults aged 50 to 75 years, resting nBRS (estimated by low frequency gain, from carotid distension rate and heart rate) and carotid stiffness were measured by high-precision carotid echotracking. PA was self-reported using the Baecke questionnaire, which distinguishes OPA, SPA, LPA and TPA. The associations between PA and nBRS and carotid stiffness were quantified using multivariate linear regression analysis. Analyses were conducted separately in the working and non- working population.

Results:
In working adults (n=5039), OPA was associated with lower nBRS function (p=0.026) and borderline higher carotid stiffness (p=0.08). The associations between OPA and nBRS remained independent after additionally adjusting for SPA (p=0.03) and exaggerated exercise blood pressure (p=0.005), a predictor of future hypertension and cardiovascular events. When examining the type of OPA separately (i.e. lifting heavy loads, standing or walking at work) lifting heavy loads only was associated with impaired nBRS (p=0.048). When stratified by education, this association remained only in those with less than tertiary education. SPA was associated with higher nBRS (p=0.0005) and borderline lower carotid stiffness (p=0.052). Neither LPA nor TPA was associated with nBRS or carotid stiffness. In non-working adults (n=3610), SPA and TPA were both associated with lower carotid stiffness (p=0.012 and p=0.020), but not nBRS. LPA was not associated with either parameter.

Conclusion:
Occupation-related PA, in particular lifting heavy loads, is associated with lower nBRS function, especially in those with lower education. Higher amounts of sport-related PA are associated with higher nBRS and lower carotid stiffness.
Abstract P176: Hypertension is Mediated by an Angiotensin II-Dependent Mechanism in Adipocyte Prorenin Receptor KO Female Mice and by the Para-Sympathetic Nervous System in Adipose Prorenin Receptor KO Male Mice
Authors: Gatineau Eva , Gong Ming , Yiannikouris Frederique
Published: December 2018

Obesity and lipodystrophy models, two opposite ends of adipose tissues dysfunction, can lead to hypertension and constitute relevant tools to decipher mechanisms involved in blood pressure control. Since sex related difference exists in blood pressure control, we aimed to determine whether the mechanism leading to hypertension in adipocyte prorenin receptor (PRR) KO mice was similar in male and female mice. Adipocyte PRR KO and control littermate (CTL) male (n=8 to 9 mice/group ) and females mice (n=6 to 7 mice/group) were fed a high fat diet. The systolic blood pressure (SBP) was evaluated by radiotelemetry. Adipose-PRR KO significantly increased SBP in both male and female mice (Day SBP, CTL male=122.2±1.9 mmHg, KO male=128.7±2.4 mmHg, P=0.046; CTL female=120.4±0.9 mmHg, KO female=125.5±1.9 mmHg, P=0.03). Acute losartan injection, an AT
1
R inhibitor, blunted elevated SBP in both CTL female and male mice. Interestingly, losartan decreased SBP to a greater extent in adipose PRR KO female mice compared with control female mice (Delta SPB, CTL male=-7.9±1.5 mmHg; KO male=-6.4±3.1 mmHg, P=0.68; CTL female=-5.3±1.9 mmHg; KO female=-14.2±0.9 mmHg, P=0.015). To assess the contribution of the autonomic nervous system, male and female mice were injected with propranolol, atropine and chlorisondamine. The tachychardic response to atropin was significantly greater in adipose-PRR KO male mice compared with control male mice (Delta Heart Rate, CTL male=+114.2±12.1 bpm; KO male=+156.0±11.1 bpm, P=0.023) but did not change in adipose-PRR KO female mice compared with control female mice (Delta Heart Rate, CTL female=+84.8±10.1 bpm; KO female=+89.8±21.3 bpm, P=0.85). In conclusion, our data strongly suggest that the elevation of SBP in adipose PRR KO mice is mediated by an AngII-dependent mechanism in female mice and by the para-sympathetic nervous system in male mice. These results support the importance of sex-specific approach for the development of personalized drugs in hypertension treatment.
Abstract P190: The Angiotensin AT
1A
Receptor Couples to G-alpha-i inAgouti-Related Peptide-ExpressingNeurons to Control Resting Metabolic Rate
Authors: Deng Guorui , Sapouckey Sarah A , Warwick Charles A , Usachev Yuriy M , Sebag Julien A , Cui Huxing , Grobe Justin L
Published: December 2018

We recently demonstrated that leptin stimulates resting metabolic rate (RMR) through a mechanism that requires angiotensin (ANG) type 1A receptors (AT
1A
) localized to neurons of the arcuate nucleus (ARC) which express Agouti-related peptide (AgRP). Genetic disruption of AT
1A
in AgRP neurons results in the loss of RMR responses to leptin, high fat diet, and various other stimuli which correlates with the disinhibition of AgRP, neuropeptide Y (NPY), and production enzymes and transporters (GAD1, GAD2, VGAT) for γ-aminobutyric acid (GABA) within the ARC. We hypothesize that AT
1A
activation in AgRP neurons causes disinhibition of AgRP, NPY, GAD1, GAD2 and VGAT expression and thus increased inhibitory neurotransmission to pre-autonomic nuclei, and we seek to understand the second-messenger network activated by AT
1A
which mediates transcriptional control of these genes. Intracellular calcium ([Ca
2+
]
i
, determined by FURA-2 fluorescence), cyclic AMP (cAMP, by LANCE assay), receptor surface localization (by HiBiT-tagging), and gene expression responses (by qPCR) to ANG were examined in immortalized mouse hypothalamic cells (N43/5 and N39 cells) that express typical markers of AgRP neurons. ANG (0.1 and 1 μM, n = 3) had no effect to modulate [Ca
2+
]
i
, but caused a dose-dependent reduction in forskolin-stimulated cAMP (EC
50
= 43 nM) which could be blocked by losartan (1 μM). ANG increased GTP-bound Gαi (PBS vehicle = 0.57±0.08, ANG 1 μM = 0.94±0.07 ratio vs IgG, p < 0.05, n = 3), reduced cell-surface localization of HiBiT-tagged AT
1A
(PBS vehicle = 0.22±0.02, ANG 0.1 μM = 0.15±0.01, p < 0.05, n = 4), and significantly (p < 0.05, n = 6) reduced AgRP and GAD1 expression by 43% and 27%, respectively. Lastly, preliminary studies suggest that pretreatment with pertussis toxin (PTX, 100 ng/mL), an inhibitor of Gαi, abrogated AgRP and GAD1 suppression by ANG (0.01 μM). Collectively these findings support the novel concept that within immortalized cells that express markers of AgRP neurons, AT
1A
couples to Gαi to reduce cAMP, which suppresses AgRP, NPY and GABA. This should disinhibit pre-motor, pre-autonomic circuits within the hypothalamus, which results in increased thermogenic sympathetic nerve activity, ultimately increasing RMR and energy expenditure.