bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 029: Epidemiological Study Evaluating Uncontrolled Systolic Blood Pressure in Patients ≥ 50 Years With Hypertension, in Relation to Stroke Risk and Current Antihypertensive Treatment Strategies (systup) - Analysis of India Cohort
Authors: Sethi Prof. Rishi , Mehta Dr. Ashwani , Kumar Dr. Viveka
Published: December 2018

Background:
Uncontrolled hypertension (HT), a critical risk factor contributing to rising burden of stroke; 57% of all stroke deaths in India. Elevated mean systolic blood pressure (MSBP) accounts for most cases of uncontrolled HT.

Aim:
To assess systolic blood pressure (SBP) in patients ≥ 50 years with uncontrolled SBP, to illustrate correlation of MSBP with stroke risk (using Stroke Riskometer
TM
), and the management of patients with uncontrolled SBP.

Methods:
SYSTUP, a non-interventional, observational study, recruited patients with age ≥50 years, with uncontrolled SBP (≥ 140 mm Hg), from 176 centers across India in real-world clinical setting. Descriptive statistics and Logistic regression analysis were used for risk calculation.

Results:
3791 patients (men 60 %) - mean age 62 years, mean BMI 27 kg/m
2
and mean BP 157/90 mmHg. A significant positive correlation was found between MSBP and stroke risk. The likelihood of 5 year and 10 year stroke risk increased by approximately 4% with 1 mmHg increase in MSBP (P <.0001). MDBP did not exhibit any significant correlation with 5 year (P=0.242) and 10 year (P=0.8038) stroke risk. Comorbid diabetes was the main risk factor of 5 year stroke risk with odds ratio of 1.6.

Majority of the patients were receiving angiotensin receptor blocker (ARB), irrespective of the risk categories - low, moderate, high, very high risk (Table).

Conclusion:
The SYSTUP India results show that elevated MSBP increases stroke risk in patients ≥ 50 years with uncontrolled HT mainly treated with ARB. These real-world clinical findings call for strengthening BP control in primary care to prevent stroke.
Abstract 039: Differential Expression of miR-146b-5p Modulates Sex-Specific Cardiac Phenotypes in a Rat Model of Chronic Kidney Disease
Authors: Paterson Mark , Kriegel Alison J
Published: December 2018

Type 4 cardiorenal syndrome is a complex disorder in which primary chronic kidney disease (CKD) contributes to secondary cardiovascular disease (CVD). Although the pathological link between the kidney and heart are well-characterized, the molecular switches which drive cardiac pathology are not well understood. miRNAs have the potential to be master regulators of cellular signaling networks through modulation of RNA and protein expression profiles and have been shown to be important in both cardiovascular health and disease. Previous work in our laboratory identified a significant increase of miR-146b-5p expression in both the heart and the kidney in a 5/6 nephrectomy (5/6Nx) model of CKD in the Sprague-Dawley rat. To evaluate how miR-146b-5p is involved with pathological changes in the heart following 5/6Nx, echocardiography, blood pressure and left ventricle (LV) pressure-volume analyses were performed on male and female wild-type (WT) and miR-146b-5p null mutant (146b
-/-
) rats for seven weeks following 5/6Nx or sham surgery. Structural analysis of the heart revealed that male and female rats exhibited significant hypertrophy of the LV wall in both the WT (+21% [1.50 vs. 1.24g] and +24% [1.00 vs. 0.81g] vs. sham, respectively; p<0.001) and 146b
-/-
(+17% [1.36 vs. 1.17g] and +26% [1.03 vs. 0.82g] vs. sham, respectively; p<0.001) genotypes. Furthermore, dilation of the LV chamber was observed in WT male (+9% LVIDd [9.45 vs. 8.67mm] vs. sham; p=0.032) and female (+29% LVIDd [9.39 vs. 7.30mm] and +36% LVIDs [5.80 vs. 4.86mm] vs. sham; p<0.005) rats 7 weeks post-5/6Nx. However, LV dilation was attenuated in 146b
-/-
females (-18% LVIDd [8.18 vs. 9.39mm] and -26% LVIDs [4.88 vs. 5.80mm] vs. WT; p=0.07) but not in 146b
-/-
males. Functionally, LV pressure-volume analysis revealed 146b
-/-
females exhibit an increase in stroke work (+59% vs. sham; p=0.063) and a significantly elevated mean arterial blood pressure (+23mmHg vs. sham; p=0.02). These alterations in cardiovascular function are not observed in miR-146b
-/-
males. Taken together, these data suggest 146b
-/-
female rats may be resistant to eccentric dilation despite increased afterload and highlight the sex-specific modulation of cardiorenal phenotypes by miR-146b-5p following 5/6Nx.
Abstract 042: Gender Differences in Response to Chronic Angiotensin-(1-7) Treatment in Obesity
Authors: White Melissa , Miller Amanda J , Loloi Justin , Bingaman Sarah , Arnold Amy C
Published: December 2018

Angiotensin (Ang)-(1-7) is a vasodilatory hormone of the renin-angiotensin system (RAS) that has recently been shown to play a role in glucose homeostasis. Our previous study showed that obese male mice are deficient in circulating Ang-(1-7), and that chronic Ang-(1-7) restoration improves insulin sensitivity independent of blood pressure. Obese female mice also develop insulin resistance and glucose intolerance; however, the ability of Ang-(1-7) to improve metabolic function in females has not been explored. We hypothesized that Ang-(1-7) would equally improve glucose homeostasis in obese female and male mice. Adult male and female C57Bl/6J mice were placed on standard chow or 60% high fat diet (HFD) for 11 weeks, with Ang-(1-7) [400ng/kg/min] or saline given subcutaneously during the last 3 weeks of diet (n=8-12/group). During the last week of treatment, we assessed body composition and performed intraperitoneal insulin tolerance (ITT) and glucose tolerance (GTT) tests. While females had lower body mass throughout the study, both genders similarly increased adiposity in response to HFD (females: 7±1 chow vs 22±1% of body mass HFD; p=0.001; males: 6±1 vs 21±2% of body mass HFD, p=0.001). Ang-(1-7) had no effect on body composition in obese males, but reduced body mass (35±1 vs 31±2 g; p=0.028) and adiposity (22±1 vs 17±2% of body mass; p=0.024) and improved lean mass (p=0.019) in obese females. Ang-(1-7) improved insulin sensitivity in both obese male and female mice (p<0.001), with no gender differences (p=0.429). Ang-(1-7) improved glucose tolerance in obese female mice [area under the curve (AUC) for change in blood glucose levels during GTT: 20748±3156 chow vs 37582±2219 HFD vs 22884±5610 HFD+Ang-(1-7); p=0.044], with no effect in obese male mice (p=0.894). There was no effect of Ang-(1-7) on any outcome in chow fed male or female mice. Our data suggests that obese female mice are more responsive to chronic Ang-(1-7) therapy in terms of metabolic outcomes compared with obese male mice. These findings may have implications for understanding gender differences in RAS mechanisms involved in obesity and related metabolic complications.
Abstract 049: Endothelial-Derived Microparticles as Biomarkers and Mediators of Endothelial Cell Injury in VEGF Inhibitor-Treated Cancer Patients: Implications in Hypertension
Authors: Neves Karla B , Rios Francisco J , McLeod Martin , Hughes Judith D , Jones Robert , Evans Jeff , Montezano Augusto , Touyz Rhian M
Published: December 2018

Vascular endothelial growth factor receptor inhibitors (VEGFRi), used as anti-angiogenic drugs to treat cancer, induce severe hypertension although the underlying molecular mechanisms are still unclear. Endothelial microparticles (MPs) are biomarkers of endothelial injury and are also functionally active since they influence downstream target cell signalling and function. We questioned whether MP status is altered in cancer patients treated with VEGFRi and whether they influence endothelial cell function associated with vascular dysfunction in hypertension. Plasma MPs were isolated from cancer patients before and after treatment with VEGFRi. Human aortic endothelial cells (HAEC) were stimulated with human plasma-isolated MPs (10
6
MPs/mL). MP characterization was assessed by flow cytometry; protein and gene expression by immunoblotting and qPCR; ROS and NO production by lucigenin and immunofluorescence. Patients treated with VEGFRi had significantly increased in endothelial cell-derived MPs (EMP) (0.19±0.02 Pre vs. 0.31±0.04 Post-treatment). HAEC exposed to post-treatment MPs increased pre-pro-ET-1 mRNA (4.10±0.97 vs. 10.57±3.54), corroborating the raise in ET-1 levels (μg/mL: 631.9±46.1 vs. 780.2±37.2) observed in HAEC stimulated with vatalanib (VEGFRi). Post-treatment MPs increased ROS generation in HAEC (100.0 vs. 5 min- 123.4±7.7, 30 min- 162.4±22.5, 60 min- 183.9±44.7), effects that were attenuated by ET
A
and ET
B
receptor blockers. VEGFRi post-treatment MPs increased phosphorylation of the inhibitory site of eNOS (Thr
495
) (100.0 vs. 181.3±18.7) and decreased NO levels in HAEC (100.0 vs. 72.7±9.9) which was inhibited by ET
B
receptor blockade (eNOS: 109.8±14.9; NO: 142.1±25.9). Gene expression of proinflammatory mediators was increased in HAEC exposed to post-treatment MPs (1.0 vs. TNF-α: 5.4±1.8, MCP-1: 2.1±0.3, iNOS: 3.1±0.8, COX2: 2.4±0.4, ICAM1: 2.7±0.6), effects inhibited by BQ123 and BQ788. In conclusion, our data identify EMPs as biomarkers of VEGFi-induced endothelial injury and important mediators of ET-1-sensitive redox-regulated endothelial cell signalling. These molecular processes may play a role in vascular dysfunction associated with hypertension in VEGFRi-treated cancer patients.
Abstract 054: Tissue-Specific Effects of Targeted Mutation of
Mir29b1
in Rats
Authors: Xue Hong , Zhang Guangyuan , Geurts Aron M , Usa Kristie , Jensen David M , Liu Yong , Liang Mingyu
Published: December 2018

miR-29 is a master regulator of extracellular matrix genes and is generally considered anti-fibrotic, and it improves nitric oxide (NO) production in human and rat arterioles by targeting Lypla1.Targeted mutation in the
Mir29b1
gene exacerbates hypertension in a rat model derived from the Dahl salt-sensitive rat. The kidney, particularly the renal outer medulla, plays a key role in the development of salt-sensitive hypertension. In the present study, we examined the effect of the targeted
Mir29b1
mutationon tissue fibrosis and NO levels with a focus on kidney regions. The abundance of miR-29b and the co-transcribed miR-29a was substantially lower in the renal cortex, renal outer medulla, heart and liver in mutant rats. In mutant rats on a 0.4% NaCl diet and prior to the development of overt hypertension, tissue fibrosis was significantly increased in the renal outer medulla, but not in the renal cortex, heart or liver, compared with wild-type littermates. Levels of NO metabolites were significantly lower, and Lypla1 protein abundance significantly higher, in the renal outer medulla, but not in the renal cortex. After 14 days of a 4% NaCl diet, tissue fibrosis remained significantly higher in the renal outer medulla and became higher in the heart of mutant rats compared with wild-type littermates, but not in the renal cortex. 24h urine volume was significantly lower in mutant rats on either the 0.4% or 4% NaCl diet. Microalbuminuria was exacerbated by the 4% NaCl diet, but was not significantly different between mutant rats and wild-type littermates. These findings indicate the effects of miR-29 are tissue-specific. The renal outer medulla might be particularly susceptible to the injurious effects of a miR-29 insufficiency, which might contribute to the development of hypertension in
Mir29b1
mutant rats.
Abstract 064: Purinergic P2X1 and P2X7 Receptors Activation Attenuate Angiotensin AT1 Receptors Dominance in Regulating the Preglomerular Renal Microcirculation in Angiotensin II Dependent Hypertension
Authors: Kulthinee Supaporn , Shao Weijian , Franco Martha , Navar L. Gabriel
Published: December 2018

Two important mechanisms regulating afferent arterioles (AA) in angiotensin II (ANG II) hypertension are the purinergic receptors (P2R) and the angiotensin AT1 receptors (AT1R); however, the nature of the interactions between the respective receptors has not been established. Accordingly, studies were performed in rats subjected to 2 weeks of ANG II infusion (80 ng/min via osmotic minipumps) which has been shown to increase interstitial ATP and ANG II concentrations. Experiments using the in vitro isolated juxtamedullary nephron preparation allowed direct visualization of the AA. To determine the interaction between P2XR and AT1R on AA at hypertensive renal perfusion pressure, afferent arteriolar inside diameters (AAD) were measured in response to increases in renal perfusion pressure (RPP) from 100 mmHg to 140 mmHg followed by superfusion with the inhibitors (i). P2X1Ri (NF-449) 1 μM, P2X7Ri (A-438079) 1 μM and P2X1Ri 1 μM plus P2X7Ri 1 μM followed by superfusion with AT1Ri (SML-1394) 1 μM. Increases in RPP to 140 mmHg decreased AAD from 14.75±0.19 μm to 11.69±0.23 μm (n=15, P<0.05) demonstrating autoregulation. Treatment with P2X1Ri significantly increased AAD from 11.94±0.54 μm to 14.33±0.15 μm (n=5, P<0.05) and increased further to 15.33±0.33 μm by treatment with the AT1Ri to values similar to those at RPP 100 mmHg (14.88±0.39 μm). Treatment with the P2X7Ri also vasodilated afferent arteriolar but to a lesser extent (13.34±0.06 μm vs. 10.94±0.21 μm, n=5, P<0.05); and further treated by AT1Ri, AAD returned to values similar to those at RPP 100 mmHg (14.05±0.20 μm vs. 14.16 ± 0.19 μm). Treatment with P2X1Ri plus P2X7Ri significantly increased AAD from 12.2±0.13 μm to 14.76±0.12 μm (n=5, P<0.05) a value not significantly different from the value at RPP 100 mmHg (15.22±0.17 μm). Further treatment with AT1Ri after treatment with the P2X1Ri plus P2X7Ri significantly increased AAD to values higher than the value observed at treatment with P2X1Ri plus P2X7Ri (15.43±0.23 μm vs. 14.76 ± 0.12 μm, P<0.05). The results indicate that renal P2X1R and P2X7R exert dominant roles in the regulation of AAD during elevation in arterial pressure and attenuate most of the AT1R influence on AAD in kidneys from ANG II hypertensive rats.
Abstract 065: Enhanced Glomerular Capillary Pressure and Tubuloglomerular Feedback (tgf) in Obese Alms1 (alstrom Syndrome 1) Knock Out Rats
Authors: Monu Sumit R , King-Medina Keyona , Ortiz Pablo
Published: December 2018

The ALMS1 gene has been associated to decreased renal function (lower GFR) and hypertension in genome-wide studies humans. The role of ALMS1 in renal dysfunction is unclear. Our lab has identified the ALMS1 protein as an interacting partner with the Na/K/2Cl cotransporter NKCC2, which mediates NaCl absorption by the thick ascending limb. We also found that rats with ALMS1 deletion exhibited increased NKCC2 activity, hypertension and progressive obesity. NKCC2 also mediates the tubuloglomerular feedback (TGF) mechanism. TGF is initiated by an increase in NaCl transport via NKCC2 in the macula densa, which causes subsequent afferent arteriolar constriction and decreases GFR. We hypothesized that ALMS1 deletion enhances TGF and reduces GFR in rats. To test this we studied homozygous ALMS1 knockout (KO) and littermates wild-type Dahl SS (WT) rats. We measured TGF using in vivo renal micropuncture technique. TGF was calculated as a reduction in stop flow pressure (P
SF
) during simultaneous perfusion of late proximal tubule incrementally from 0 to 20 and then 40 nl/min. P
SF
is an index of glomerular capillary pressure (P
GC
) and increased P
GC
is linked to renal dysfunction. Increasing the perfusion rate from 0 to 20nl/min decreased P
SF
in ALMS1 KO rats but not in WT rats, indicating enhanced TGF sensitivity (ALMS1: 45.5±2.9 to 41.4±3.0 mmHg, p<0.05 vs. WT: 34.2±1.4 to 31.4±1.9 mmHg, N.S). Baseline P
GC
was elevated (ALMS1: 46.9±3.3 vs. WT: 34.9±1.5 mmHg, p<0.05) while renal interstitial hydrostatic pressure (RIHP) was reduced in ALMS1 KO (ALMS1: 2.6±0.9 vs. WT: 5.2±0.5 mmHg, p<0.05). We then measured GFR (calculated from inulin clearance) and found it was reduced in ALMS1 KO rats (ALMS1: 0.5±0.1 vs. WT: 1.6±0.3 ml/min/100gBW, p<0.05). Mean arterial pressure was higher in ALMS1 KO rats (ALMS1: 146±2 vs. WT: 113±3 mmHg, p<0.05). We concluded that ALMS1 is involved in the control of glomerular hemodynamics. Deletion of ALMS1 enhanced TGF sensitivity and this may be in part responsible for decreased GFR. It is likely that increased TGF-sensitivity, enhanced glomerular capillary pressure and hypertension combine to reduce GFR in ALMS1 KO rats. These are the first data supporting a role of ALMS1 in glomerular hemodynamics.
Abstract 073: Sleep Duration and 24-hour Ambulatory Blood Pressure in Adults not on Antihypertensive Medications
Authors: Cohen Jordana B , Shulman Rachel , Cohen Debbie L , Grandner Michael A , Gislason Thorarinn , Pack Allan I , Kuna Samuel T , Townsend Raymond R
Published: December 2018

Background:
Short sleep duration has been widely linked to increased cardiovascular morbidity and mortality, although the underlying mechanisms for this relationship remain unclear. Maladaptive circadian variations in blood pressure (BP) may be an important mediator of adverse outcomes in people with short sleep duration. Our goal was to evaluate the association between sleep duration and 24-hour BP control.

Methods:
We performed a post-hoc analyses of 24-hour ambulatory BP monitoring in 219 non-hypertensive and pre-hypertensive adults in the Lifestyle Modification in BP Lowering Study (LIMBS) and Penn Icelandic Sleep Apnea (PISA) Study.

Results:
The 24-hour mean systolic BP was 12.7 mmHg higher in LIMBS (see Figure; p<0.001) and 4.7 mmHg higher in PISA (p=0.005) among participants with shorter sleep duration (<7 hours) compared to those with longer sleep duration (≥7 hours). There was a 1 mmHg higher 24-hour mean systolic BP for every 2.57 minute shorter sleep duration in LIMBS (95% CI -4.56 to -0.58; p<0.001), and a 1 mmHg higher 24-hour mean systolic BP for every 1.99 minute shorter sleep duration in PISA (95% CI -3.48 to -0.50; p=0.009). There was no significant association between sleep duration and BP variability, nocturnal dipping, or in-office systolic BP.

Conclusions:
Shorter sleep duration is strongly associated with higher systolic BP on 24-hour ambulatory BP monitoring; the association seems to be independent of nocturnal and in-office BP. Adults with shorter sleep duration may benefit from screening with 24-hour ambulatory BP monitoring to promote earlier detection of hypertension and potentially mitigate their increased risk for future cardiovascular disease.
Abstract 085: Renal and Neural Mechanisms of Age-Related Hypertension
Authors: Frame Alissa A , Wainford Richard D
Published: December 2018

Aim:
Hypertension (HTN) is positively correlated with age and sympathetic tone in humans. We hypothesize that sympathetically driven sodium chloride cotransporter (NCC)-mediated sodium retention contributes to age-related HTN.

Methods:
Three, 8 and 16 month old male SD rats on a normal salt (NS; 0.6% NaCl) diet underwent an acute IV volume expansion (VE; 5% body weight – mechanoreceptor stimulus) or 1M NaCl infusion (20μL/min, 2hr – chemoreceptor stimulus) and mean arterial pressure (MAP), natriuresis (UNaV) and paraventricular nucleus (PVN) neuronal activation (c-Fos expression) were assessed. In separate groups of rats fed a 21 day NS or high salt (HS; 4% NaCl) diet, 1) ex vivo afferent renal nerve (ARN) activity (norepinephrine-evoked substance P release) or 2) in vivo MAP, NCC activity (ΔUNaV to IV hydrochlorothiazide, 2mg/kg) and sympathetic tone were assessed. N=4-6/gp.

Results:
The natriuretic and PVN sympathoinhibitory parvocellular neuronal responses to VE, but not 1M NaCl infusion, are blunted in aged rats. Aged rats exhibit reduced ARN activity on a NS diet and fail to increase ARN activity on a HS diet. Aged rats develop salt sensitive HTN accompanied by elevated NCC activity and sympathetic tone on a NS diet and impaired suppression of NCC activity and sympathetic tone on a HS diet.

Conclusion:
Aging is associated with a selective impairment in the mechanosensitive ARN and salt sensitive HTN. We speculate that an age-related decrease in the mechanosensitive ARN sympathoinhibitory reno-renal reflex promotes sympathoexcitation, perhaps via reduced activity of PVN sympathoinhibitory parvocellular neurons, driving NCC-mediated sodium retention and salt sensitive HTN.
Abstract 086: Neuroanatomical Basis of PVN MC4R-Expressing Neurons for Sympathetic Cardiovascular Control
Authors: Singh Uday , Toth Brandon A. , Davis Kevin C. , Saito Kenji , Morgan Donald A. , Rahmouni Kamal , Cui Huxing
Published: December 2018

It is well established that the central melanocortin system is critical for autonomic functions and energy homeostasis mainly via signaling at melanocortin-4 receptor (MC4R). Importantly, while obesity is commonly associated with elevated sympathetic tone and blood pressure, severely obese humans and rodents due to genetic MC4R deficiency exhibit normal to low sympathetic tone and blood pressure, suggesting a significant role of MC4R pathway in mediating obesity-associated sympathoexcitation and hypertension. MC4R is widely expressed in the brain including hypothalamic paraventricular nucleus (PVN) which regulates feeding and sympathetic traffic. However, the neuroanatomical basis of PVN MC4R neurons for sympathetic regulation is unclear. The goal of current study is to map the PVN MC4R neural circuits affecting sympathetic tone in mice. To this end, we injected Cre-dependent AAV driving eYFP-fused channel rhodopsin-2 expression into the PVN of MC4R-t2a-Cre knock-in mice, which allow targeted anterograde tract-tracing of PVN MC4R neurons throughout the brain. In addition to known brain region for feeding behavior (i.e. parabrachial nucleus), we found broad innervations of PVN MC4R neurons to various brain regions important for autonomic-cardiovascular control, including, but not limited to, nucleus of solitary tractus, dorsomotor nucleus of vagus, and ventrolateral medulla. Considerable innervation was also evident in spinal cord, which is further confirmed by Fluoro-gold (FG)-mediated retrograde tracing in the spinal cord (thoracic T6-10) of MC4R-GFP transgenic mouse. Double immunofluorescence labeling of GFP and FG revealed that ~50% (58 out of 116) MC4R neurons in the posterior parvocellular subdivision of PVN project to thoracic spinal cord. Furthermore, microinjection of synthetic MC4R agonist (MTII) into the PVN evokes ~42% increase (from baseline) in renal sympathetic nerve activity in anesthetized mice. These results provide important insights into understanding the divergent neural circuits by which PVN MC4R signaling differentially regulates metabolic and cardiovascular functions. Functional dissection of these diverging neural pathways using optogenetic/chemogenetic approaches is ongoing.
Abstract 088: The Immune Mechanisms of Salt-Sensitivity
Authors: Ruggeri Barbaro Natalia , Van Beusecum Justin , Loperena Roxana , Alsouqi Aseel , Ao Mingfang , Elijovich Fernando , Laffer Cheryl L , Ikizler Alp , McDonough Alicia A , Moreno Heitor , Harrison David G , Kirabo Annet
Published: December 2018

Salt-sensitivity is present in 50% of all hypertensive individuals. Prior studies have focused on the roles of kidney, vasculature and sympathetic activity in salt-sensitivity but the contribution of immune cells is poorly understood. We recently found that in murine dendritic cells amiloride sensitive channels sense salt and trigger NADPH oxidase-dependent formation of isolevuglandin-(IsolG)-adducts. We tested the hypothesis that human monocytes exhibit salt-sensitivity leading to activation via IsoLG-adduct formation and this is associated with cardiovascular risk factors. In a cohort of 18 subjects, we found that the sodium intake, measured by 24 hours urine excretion (UNa) was positively correlated with plasma levels of IsolGs. We also measured accumulation of interstitial sodium in 70 subjects by Magnetic Sodium Resonance and evaluated their circulating monocytes by flow cytometry. Subjects with high skin sodium had higher levels of IsoLGs in their monocytes (24±6 vs 38±6 %, p<0.05) and higher expression of CD83, an activation and dendritic cell marker (0.04 ± 0.009 vs 0.12± 0.04%,
p
=0.04). To investigate the ability of monocytes to respond to salt, we cultured monocytes from 17 subjects in high salt environment (HS:190 mM NaCl) or normal media (NS:150mM NaCl) for 48 hours. In culture, 47% of the subjects respond to salt, denoted by an increase of at least 20% in IsoLG formation (NS: 1327±240
vs.
HS: 2217±653,
p
=0.009) as well as increased expression of the activation markers CD83 and CD86. The subjects’ cardiovascular risk factors including pulse pressure, BMI, glucose and total cholesterol positively correlated with the amount of IsolGs produced (ΔHS-NS) in response to salt (r=0.51 p<0.05, r=0.66 p=0.005, r=0.55 p<0.05, p=0.72 p=0.003, respectively). Interestingly, 5 pM of Ouabain, a Na-K-ATPase blocker, increased intracellular sodium and expression of CD86 (NS: 98±11, HS: 203 ±16 vs, NS+ Ouabain: 476 ± 58 MFI, p=0.001) and CD83 (NS: 778±90, HS: 1529 ± 94 vs, NS+ Ouabain: 1649 ± 209 MFI, p=0.003). We suggest that in addition to the kidney and vasculature, human monocytes and monocyte derived cells exhibit salt sensitivity, and that this is conveyed by cardiovascular risk factors and activity of the Na-K-ATPase.
Abstract 101: Transforming Growth Factor-Beta-Receptor Antagonist Blocks the Cardiac Fibrosis and Remodeling in Guanylyl Cyclase/Natriuretic Peptide Receptor-A Gene-Disrupted Mice
Authors: Subramanian Umadevi , Samivel Ramachandran , Zhao Hanqing , Gogulamudi Venkateswara , Pandey Kailash N
Published: December 2018

Targeted-disruption of guanylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) gene (
Npr1
) exhibits hypertension and provokes congestive heart failure in mice; however, the underlying mechanisms are not well clear. The objective of this study was to determine whether transforming growth factor-beta receptor (TGF-βR) antagonist, GW788388 inhibits the development of cardiac fibrosis and remodeling in
Npr1
gene-disrupted mice. The adult male (16-20 weeks)
Npr1
null mutant (
Npr1
-/-
, 0-copy), heterozygous (
Npr1
+/-
, 1-copy), and wild-type (
Npr1
+/+
, 2-copy) mice were orally administered with TGF-βR antagonist, GW788388 (1 mg/kg/day) for 28 days. The expression of cardiac fibrotic markers was analyzed using real-time PCR and Western blot. Heart weight-to-body weight (HW/BW) ratios were determined and heart functions were measured by echocardiographic analysis. The
Npr1
-/-
mice showed markedly increased cardiac fibrosis and HW/BW ratio with increased expression of collagen-1α (3.5-fold), monocyte chemoattractant protein (4-fold), connective tissue growth factor (CTGF, 5-fold), α-smooth muscle actin (α-SMA, 4-fold), TGF-βRI (4-fold), TGF-βRII (3.5-fold) and SMAD proteins (SMAD-2, 5-fold; SMAD-3, 3-fold) compared with
Npr1
+/+
mice. The expression of phosphorylated extracellular-regulated kinase (pERK1/2) was also up-regulated by 68% (
P
<0.001) in

Npr1
-/-

mice. The treatment of

Npr1
-/-

mice with GW788388 prevented the development of cardiac fibrosis and down-regulated the expression of fibrotic markers and SMAD proteins by 70-75% (
P
<0.001) compared to vehicle-treated mice. In contrast, the expression of pERK1/2 proteins was unaffected in GW7885388-treated mice excluding the involvement of non-genomic pathway. The left ventricular dimensions (systole and diastole) and fractional shortening were significantly (
P
<0.001) improved in the drug-treated

Npr1
-/-

mice. The results suggest that the cardiac fibrosis, remodeling, and dysfunction in
Npr1
-/-
mice are regulated through TGF-βR-mediated SMAD-dependent canonical pathway. The findings will be important for the development of new molecular therapeutic targets for the treatment of cardiac fibrosis, remodeling, and dysfunction in humans.
Abstract 115: Renal-Specific Expression of Human Grk4 R65L in Mice Lacking Grk4 Increases Blood Pressure
Authors: Rozyyev Selim , Konkalmatt Prasad , Asico Laureano , Kumar Megha , Hunt Jessica , Jose Pedro A , Armando Ines
Published: December 2018

The genetic causes of salt (NaCl) sensitivity in humans are not well understood. The kidney is critical to the overall fluid and electrolyte balance and long-term regulation of blood pressure (BP). Therefore, the pathogenesis of salt sensitivity must involve a deficient renal NaCl handling. The renal paracrine inhibition of Na
+
transport by dopamine is impaired in salt-sensitive rats, mice, and humans due to decreased dopamine D1 receptor (D1R) function and not related to a primary defect of the D1R gene but rather due to its uncoupling from second messengers caused by activating variants of the human G protein-coupled receptor kinase type 4 (h
GRK4
) (R65L, A142V, and A486V). We have shown that global transgenic mice expressing hGRK4 R65L variant have salt-sensitive hypertension. To determine the role of the kidney in the salt-sensitivity of these mice, we developed a mouse model of kidney-specific expression of the R65L variant (KS hGRK4 R65L) by bilateral ureteral infusion of AAV vectors carrying the variant in GRK4 knockout mice. Mice infused with hGRK4 wild-type served as controls (KS hGRK4 WT). Systolic BP (SBP) was measured under anesthesia in the mice before and after infusion of the AAV vectors. SBP before AAV was similar in both groups. SBP post AAV increased in KS hGRK4 R65L (93±1 vs 117±4 mmHg, P<0.05, n=4) but not in KS hGRK4 WT (96±2 vs 105±6, n=5). Renal mRNA expressions of dopamine 1 (D1R, 0.81±0.01 vs 1.28±0.04, P<0.01), D3R (0.44±0.02 vs 1.27±0.07, P<0.01) and angiotensin AT1A (AT1A, 0.78±0.02 vs 1.02±0.02, P<0.01) receptors were decreased in renal cortex of KS hGRK4 R65L relative to KS hGRK4 WT. In addition, the expressions of Na
+
/K
+
ATPase (1.12±0.03 vs 1.0 ±0.007, P<0.05) and ENaC (1.4±0.14 vs 1.0±0.11, P<0.05) were increased while those of the proximal tubule Na
+
transporters NaPi2 (0.81±0.02 vs 1.04±0.02), SGLT2 (0.89±0.03 vs 1.07±0.05), and NBCe2 (0.50±0.07 vs 1.15±0.03) were decreased. Our results show that kidney specific expression of hGRK4 R65L increased SBP and produced changes in the expression of receptors and Na
+
transporters, pump, and channel but their roles in the increased SBP remain to be determined.
Abstract 122: Deletion of
Gstm1
Results in Poor Survival and Exaggerated Kidney Injury Associated With Increased Oxidative Stress and Apoptosis in a Mouse Model of Chronic Kidney Disease
Authors: Cechova Sylvia , Bodonyi-Kovacs Gabor , Chan Fang , Ruiz Phillip , Le Thu H.
Published: December 2018

In humans, a common deletion variant of the
GSTM1
gene, the
GSTM1(0)
null allele, results in decreased GSTM1 enzymatic activity and is associated with higher levels of oxidative stress. We reported that
GSTM1(0)
is associated with increased risks of chronic kidney disease (CKD) progression and death in the African Americans Study of Kidney Disease. To establish a causal relationship between GSTM1 deficiency and CKD progression, we deleted
Gstm1
in the mouse, and determined its effect in the sub-total nephrectomy (Nx) model of CKD. After the 8
th
week after Nx, 4 out of 8
Gstm1
knockout (KO) mice began to die, whereas wild-type (WT) mice had 100% survival by the study endpoint at 13
th
week. Remaining KO mice had significantly higher plasma creatinine (p < 0.05), suggestive of lower GFR. Due to early mortality in KO mice after the 8th week after Nx, we phenotyped mice at the 8
th
week (n=6 each). KO mice had significantly higher 1) mean systolic blood pressure (by radiotelemetry): KO 157.2 ± 5, WT 141.1 ± 2.4 mmHg, 2) urine albumin/creatinine ratio (μg/mg): KO 748.4 ± 187.9, WT 235.1 ± 24.6, and 3) renal superoxide (O2·-) levels (counts/min/mg dry tissue): KO 249.7 ± 63; WT 74.5 ± 14.7; P ≤ 0.02. Hydrogen peroxide levels were not different. Renal mRNA levels of the superoxide dismutase isoforms SOD1, SOD2, and SOD3 were significantly lower in KO mice (KO 7.78 ± 0.047, 0.446 ± 0.062, 0.18 ± 0.03; WT 10.64 ± 0.43, 2.432 ± 0.386, 1.188 ± 0.124, respectively, P ≤ 0.003. Total kidney pathology score (KO 18.7 ± 1.5; WT 8 ± 0.6; P <0.0001) showed that KO mice had worse kidney injury, with severe glomerulosclerosis, proteinaceous casts, and chronic inflammation. Both glomerular surface area (SA)/total non-infarcted kidney area (KO 3.347 ± 0.301; WT 2.286 ± 0.268; P = 0.02) and mesangial SA/glomerular SA (KO 35.62 ± 2.219; WT 23.6 ± 2.569; P = 0.005) were significantly increased in KO mice. GSTM1 has also been shown to suppress ASK1 - the apoptosis signal-regulating kinase 1 that induces apoptosis via activation of caspase-3. Consistent with activated ASK1 pathway, KO mice had significantly higher renal TUNEL staining and caspase-3 protein levels (KO 3.594 ± 0.327; WT 2.511 ± 0.208; P = 0.01). In conclusion, in CKD, loss of GSTM1 leads to deleterious outcome associated with increased O2·- levels and apoptosis.
Abstract 124: Nadph Oxidase 4 and Mitochondrial Abnormalities Contribute to Oxidative Stress and Endothelial Dysfunction in Young Normotensive Patients With Autosomal Dominant Polycystic Kidney Disease
Authors: Kahveci Ali , Chebib Fouad T , Eirin Alfonso , Lerman Amir , Lerman Lilach O , Harris Peter C , Torres Vicente E , Irazabal Maria V
Published: December 2018

Background:
Fifty percent of patients with autosomal dominant polycystic kidney disease (ADPKD) present hypertension (HTN) early on, which increases to nearly 100% at end-stage, and is associated with faster renal functional decline. Endothelial dysfunction (ED) accompanied by oxidative stress (OS) precedes the development of HTN, but the mechanisms responsible remain unknown. The aim of this study was to determine whether NADPH oxidase (NOX4) and mitochondrial dysfunction contribute to OS and ED preceding HTN in early ADPKD.

Methods:
We prospectively measured plasma levels of homocysteine (Hcy), 8-isoprostane, NOX4 and the mtDNA genes COX3 and ND1 in young normotensive (without BP medication) ADPKD patients, and age-matched healthy volunteers (HV) (n=10, each). Total kidney volume (TKV) and Renal Blood Flow (RBF) were evaluated by MRI.

Results:
BP levels were higher in ADPKD, and HtTKV was twofold higher in ADPKD vs. controls. eGFR and RBF were similar between the groups (Table). Plasma Hcy, 8-isoprostanes and NOX4 were higher in ADPKD, and correlated directly with HtTKV (p<0.05, Figure), but the correlation with RBF was not significant. Plasma mtDNA copy number were lower in ADPKD compared to controls, and correlated inversely with HtTKV(R
2
0.397 and 0.392 respectively).

Conclusion:
Early ADPKD is associated with elevation in Hcy, 8-isoprostane, and NOX4 levels, and decreased mtDNA copy numbers, which precede the reduction in RBF and the development of HTN, and are associated with disease severity. NOX4 and mitochondrial abnormalities may contribute to oxidative stress, endothelial dysfunction, and possibly the development of HTN and disease progression in ADPKD.
Abstract 125: Hyperacetylation of CypD Contributes to Mitochondrial Dysfunction, Vascular Oxidative Stress and Hypertension, and Mitochondria-Targeted Isoketal Scavenger mito2HOBA Prevents CypD Hyperacetylation and Reduces Hypertension
Authors: Dikalov Sergey I , Mayorov Vladimir , Pandey Arvind , Panov Alexander , Itani Hana , Amarnath Venkataraman , Mernaugh Raymond , Boutaud Olivier , Davies Sean , Oates John , Harrison David G , Dikalova Anna
Published: December 2018

One third of adult population has hypertension and third of hypertensive patients remain hypertensive despite treatment with multiple drugs. There have been no mechanistically novel treatments for hypertension in the past 30 years. We propose that new agents targeting CypD activation, a regulatory subunit of mitochondrial permeability transition pore, could add to the currently available therapeutic armamentarium to improve treatment of hypertension. To test this hypothesis we have studied CypD acetylation which was associated with CypD activation. We found that angiotensin II-induced hypertension is linked to CypD-K166 hyperacetylation, mPTP opening, impaired mitochondrial respiration and reduced kidney ATP. CypD hyperacetylation may result from imbalance between GCN5L1 acetyltransferase and reduced Sirt3 deacetylase activity. Indeed, aortic mitochondrial level of GCN5L1 increased by 50% while Sirt3 was reduced by 40% in angiotensin II infused mice. Sirt3 can be inactivated by cytotoxic reactive lipid dicarbonyls derived from arachidonic acid, isoketals, leading to mitochondrial hyperacetylation. To test the role of mitochondrial isoketals we synthesized a novel mitochondria-targeted isoketal scavenger mito2HOBA. mito2HOBA supplementation in drinking water (0.1g/Liter) attenuates angiotensin II-induced hypertension, prevents accumulation of mitochondrial isoketal protein adducts in the aorta and kidney measured by LC/MS and D11 antibody, improves Sirt3/GCNL1 ratio, normalizes the CypD acetylation, attenuates mPTP opening, protects mitochondrial respiration and preserves normal kidney ATP level. Decreased NO bioavailability is a hallmark of endothelial oxidative stress in hypertension due to superoxide overproduction and NO oxidation. Interestingly, angiotensin II increases vascular superoxide by 2-fold and reduces endothelial NO level to 50% while mito2HOBA diminishes vascular O
2

•

and preserves NO bioavailability in angiotensin II-infused mice. These data support the role of isoketals-mediated CypD hyperacetylation in endothelial dysfunction and hypertension. We conclude that scavenging of mitochondrial isoketals may have therapeutic potential in treatment of hypertension and target-organ-damage.
Abstract 129: TRPM7 α-kinase Deficiency Causes Cardiovascular Inflammation and Fibrosis.
Authors: Rios Francisco J , Zhi-guo Zou , Camargo Livia L , Harvey Adam P , Lacchini Silvia , Anyfanti Panagiota , McGrath Sarah , Goodyear Carl S , Montezano Augusto C , Touyz Rhian M
Published: December 2018

We previously demonstrated that TRPM7, a Mg
2+
/cation channel fused to an α-kinase domain, is regulated by vasoactive mediators and plays a protective role in hypertension. Here we questioned whether TRPM7-kinase influences vascular inflammation and fibrosis. We used Wild-type (WT) and heterozygote mutant mice for TRPM7-kinase (M7+/-). Vascular inflammatory responses were assessed
ex vivo
by intravital microscopy. Immune cells were investigated by flow cytometry. Fibrosis was investigated by sirius-red staining. Bone-marrow derived macrophages (BMDM) and Cardiac fibroblasts (CF) were obtained from WT and M7+/. [Mg
2+
]i in cardiac tissue, cardiac macrophages and circulating monocytes was significantly reduced (30-50%) in M7+/- vs WT mice. In small arteries studied by intravital microscopy, leukocytes from M7+/- showed reduced velocity (47%), increased adhesion (222%) and transmigration (480%). Expression of vascular pro-inflammatory markers including VCAM-1(33-fold), iNOS (12-fold), and IL-12 (6.8-fold) was increased in M7+/- vs WT. Cardiac galectin-3 (Gal-3) levels (16.6±3.6 vs WT 9.2±1.2 cells/field), collagen area (6.7% vs WT 2.6%), infiltration of CD45+ cells (6±0.6% vs WT 4±0.4%) and protein expression of fibronectin (280%), TGFβ (125%), and p-Smad3 (66%), were increased in M7+/- mice. BMDM macrophages from M7+/- exhibited increased levels of Gal-3 (2.6±0.05 vs WT 2.1±0.09ng/mL), IL-10 (807±92 vs WT 305±37 pg/mL) and IL-6 (84±8 vs WT 13±5 pg/mL). A similar profile was demonstrated in resident peritoneal macrophages. CF treated with supernatant of macrophages from M7+/- increased fibronectin (43%) and PCNA (36%) vs WT. To evaluate whether these processes are Mg
2+
-sensitive, we examined effects of Mg
2+
treatment and demonstrated that Mg
2+
ameliorated pro-fibrotic and pro-inflammatory signalling evident in TRPM7+/- mice. In conclusion, TRPM7-kinase deficiency is associated with cardiac and vascular inflammation and fibrosis, processes associated with cellular Mg
2+
deficiency. Our findings highlight an important cardiovascular protective role of TRPM7 and Mg
2+
.
Abstract 131: Microbiota-Derived Metabolite Propionate Protects From Hypertensive Cardiovascular Damage
Authors: Wilck Nicola , Bartolomaeus Hendrik , Balogh András , Yakoub Mina , Homann Susanne , Markó Lajos , Höges Sascha , Tsvetkov Dmitry , Wundersitz Sebastian , Haase Nadine , Hering Lydia , Maase Martina , Kusche-Vihrog Kristina , Grandoch Maria , Fielitz Jens , Kempa Stefan , Forslund Sofia K , Kushugulova Almagul , Dechend Ralf , Eckardt Kai-Uwe , Rump Lars C , Müller Dominik N , Stegbauer Johannes
Published: December 2018

Objective:
Inflammation drives cardiovascular disease, anti-inflammatory approaches may be beneficial. Short-chain fatty acids (SCFA) are bacterial metabolites with anti-inflammatory properties affecting host immune homeostasis including regulatory T cells (Treg). We investigated effects of the SCFA propionate (administered in drinking water, NaCl as control) in two mouse models, namely hypertensive heart disease (wild-type NMRI (WT), angiotensin (Ang)II infusion 1.44 mg/kg/d s.c. for 14 days) and atherosclerosis (Apolipoprotein E knockout (ApoE), AngII infusion 0.72 mg/kg/d s.c. for 28 days), respectively.

Results:
Propionate attenuated cardiac hypertrophy and fibrosis in both models significantly. Susceptibility to cardiac ventricular arrhythmias was significantly reduced in propionate-treated WT mice. Aortic atherosclerotic lesion area was significantly reduced in propionate-treated ApoE (27.6±8 vs. 7.9±2.4%). Treatment reduced splenic effector memory (CD4+ CD44+ CD62L-) T cell frequencies (WT: 30.5±4.6 vs. 19.1±1.6; ApoE: 41.1±3.1 vs. 32.7±1.4%) and splenic Th17 cells (WT: 1.0±0.2 vs. 0.6±0.1; ApoE: 1.3±0.1 vs. 0.9±0.1%) in both models, indicating beneficial effects on systemic inflammation. Similarly, propionate reduced cardiac immune cell infiltration (CD4+, CD8+, F4/80+) in WT mice. Propionate improved vascular dysfunction and moderately reduced blood pressure in both models. Organ-protective actions of propionate (cardiac inflammation and fibrosis) were abrogated in Treg-depleted (antiCD25-treated) AngII-infused WT mice, suggesting a central role for Treg. To verify our findings in a human cohort, we re-analyzed clinical and metagenomic data from a recent randomized controlled trial investigating the effect of a 90-day synbiotic intervention in 84 subjects with metabolic syndrome including healthy controls. Interestingly, in synbiotic-treated subjects an increased capacity for SFCA production was significantly correlated to blood pressure reduction.

Conclusion:
Data underscore the importance of SCFA for cardiovascular health and suggest that lifestyle modifications leading to augmented SCFA production could be a beneficial non-pharmacological add-on strategy for cardiovascular disease.
Abstract 135: Impaired Autonomic Function in Young Adults Born Preterm With Very Low Birth Weight is Associated With Elevated Serum Uric Acid Levels
Authors: Shaltout Hossam A , Nixon Patricia A , Chappell Mark C , Diz Debra I , South Andrew M , Jensen Elizabeth T , Washburn Lisa K
Published: December 2018

Preterm birth increases the risk of cardiometabolic disease. We showed that prematurity induces programming effects that lead to elevated blood pressure (BP), higher serum uric acid, and impaired autonomic control characterized by reduced heart rate variability (HRV), baroreflex sensitivity (BRS), and elevated ratio of low frequency to high frequency power (LF/HF), a marker of sympathovagal balance. However, the relationships among these factors in subjects born preterm are undefined. Since uric acid has been shown to correlate with HRV in adult hypertensive subjects, we hypothesized that higher uric acid is associated with impaired autonomic function in subjects born preterm.

Methods:
A cohort of 131 young adults born preterm was compared to a cohort of 26 born term (19.6 years). Serum uric acid was measured. BP and ECG were recorded continuously for analysis of autonomic function reflected in HRV and BRS. We used generalized linear models to estimate the association between uric acid and measures of HRV and BRS, adjusting for race, age at follow up, and BMI. The potential interaction between uric acid and preterm birth was tested by introducing an interaction term and, if suggestive of an interaction (p ≤ 0.1), estimates were calculated within strata (preterm & term).

Results:
Compared to term, preterm participants had higher serum uric acid levels (5.2 mg/dL, IQR: 4.2, 6.2 vs 4.8 mg/dL, IQR: 3.9, 6.0) and lower HRV as measured by standard deviation of normal to normal RR intervals (71 ms, IQR: 54, 97 vs 80 ms, IQR: 55, 101). Across term and preterm groups combined, uric acid was inversely associated with BRS measured as HF alpha (
β
: -0.12, 95% CI -0.20 to -0.03, p for interaction = 0.06). Assessment of interaction showed this association was significant in preterms only (
β
: -0.15, 95% CI -0.24 to -0.06 vs 0.07, -0.17 to 0.30). Uric acid was positively associated with LF/HF (
β
: 0.13, 95% CI 0.01 to 0.25, p for interaction=0.01).

Conclusions:
Young adults born preterm had dampened parasympathetic control of heart rate and BP, and higher serum uric acid was associated with sympathovagal imbalance. The data suggest that alterations of uric acid metabolism associated with preterm birth may increase the risk of early cardiovascular disease in part by impairing autonomic control
Abstract P127: Pericyte Detachment is Involved in Renal Congestion in a Novel Rat Model
Authors: Shimada Satoshi , Hirose Takuo , Takahashi Chika , Sato Emiko , Kinugasa Satoshi , Ito Sadayoshi , Mori Takefumi
Published: December 2018

Increased central venous pressure in congestive heart failure is responsible for renal dysfunction; however, the underlying mechanisms are unclear. We hypothesized that renal interstitial hydrostatic pressure (RIHP) and expansion pressures of the vasa recta are responsible for pericyte detachment resulting renal congestion-mediated fibrosis. We created a novel rat renal congestion model and investigated the effect of renal congestion on hemodynamics and its molecular mechanisms. The inferior vena cava (IVC) between the renal veins was ligated by suture in male Sprague-Dawley rats to increase upstream IVC pressure and induce congestion in the left kidney only. Left kidney congestion reduced renal blood flow in cortex (33.6%, 28.3 to 16.0 mL/min) and in medulla (41.8%, 11.9 to 6.9 mL/min) and glomerular filtration rate (1.16 to 0.20 mL/min/kg BW), and increased RIHP (12.6 to 17.6 mm Hg). Hypoxia was observed in the medullary thick ascending limb of Henle, only in the congestive kidneys. Tubulointerstitial injury, podocyte injury, albuminuria, and reduced creatinine clearance were observed in the congestive kidneys. Molecules related to extracellular matrix expansion, tubular injury, and focal adhesion were upregulated in microarray analysis. Renal decapsulation ameliorated the tubulointerstitial injury (mRNA expression of
Kim1
15.3 to 4.3 A.U.,
αSma
5.3 to 2.8 A.U.). Electron microscopy captured pericyte detachment in the congestive kidneys. Transgelin and platelet-derived growth factor receptors (PDGFRs), as indicators of pericyte-myofibroblast transition, were upregulated in the pericytes and the adjacent interstitium. Imatinib, a PDGFR inhibitor, ameliorated the interstitial injury. Our results reveal a novel mechanism of worsening renal function associated with congestive heart failure, and provide a new therapeutic strategy based on a better understanding of the pericyte-myofibroblast transition.