bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 424: Cardiac Hypertrophy In Angiotensin Ii-dependent Hypertension: Dominant Effect Of Blood Pressure
Authors: Sparks Matthew A , Stegbauer Johannes , Chen Daian , Vivekanandan-Giri Anuradha , Pennathur Subramaniam , Crowley Steven D , Gurley Susan B , Coffman Thomas M
Published: March 2022

Left ventricular hypertrophy (LVH) is a common manifestation of end-organ injury in HTN. ACEi and ARBs attenuate the development of LVH through actions suggested to depend on direct interruption of AT1R in cardiac myocytes, rather than their effects to lower BP. In order to define the contribution of AT1R in cardiac myocytes to LVH, we made 2 mouse lines with cell-specific deletion of AT1AR from the heart using a floxed Agtr1a allele. In the 1st line, loss of AT1AR was achieved using a heterologous Cre transgene driven by the Sm22 promoter (Tagln-Cre) expressing Cre in VSMC and cardiac myocytes. There was virtually complete loss of AT1AR from cardiac myocytes and VSMCs in aorta, but not resistance vessels. Thus, with chronic AngII infusion, Tagln-KOs develop HTN to the same level as controls (MAP: 157±6 vs. 153±6 mm Hg, P=NS). In AngII-infused controls, there was cardiac oxidative stress indicated by enhanced nitrated tyrosine, but this was reduced by ≈75% in Tagln-KOs (P<0.005). Despite the absence of AT1AR in cardiac myocytes and differences in cardiac free radicals, the extent of cardiac hypertrophy was similar in Tagln-KOs and controls (6.9±0.2 vs. 6.7±0.2 mg/gm; P=NS). Similarly, myocardial BNP mRNA levels and the severity of cardiac injury were not reduced in Tagln-KOs compared to controls. Hence, the phenotype and molecular signature of LVH were not affected by the absence of cardiac AT1AR in Tagln-KOs. The 2nd mouse line of mice utilized a Cre cassette “knocked-in” to the Sm22 locus (KISm22-KOs). Like the Tagln-KOs, the KISm22-KOs exhibited elimination of AT1AR from cardiac myocytes and VSMCs in large arteries, but in this case AT1AR were also deleted from VSMCs in resistance vessels. Accordingly, Ang II-induced HTN was attenuated in KISm22-KOs (MAP: 122±3 vs. 145±4 mm Hg; P=0.004). Moreover, unlike the Tagln-KOs, cardiac hypertrophy was also diminished (5.5±0.2 vs. 7.4±0.6 mg/gm; p=0.01) and BNP mRNA levels were reduced (1.0±0.2 vs 2.6±0.8; p<0.05) compared to controls. In summary, in two models lacking AT1AR in the heart, the extent of cardiac hypertrophy in AngII HTN followed the BP, rather than AT1AR expression in cardiac myocytes. Our findings suggest the magnitude of BP elevation is of primary importance in driving the development of cardiac hypertrophy.
Abstract 429: PTP1B, a Link Between Endoplasmic Reticulum Stress and Endothelial Dysfunction
Authors: Belin de Chantemele Eric J , Anderson Ruchi , Norman Joseph B , Tremblay Michel L , Huby Anne-Cecile E , Patel Vijay S
Published: March 2022

Endothelial dysfunction is a common feature of diabetes and a major contributor to diabetic vascular complications. Despite aggressive diabetes management strategies, preventing diabetes-induced endothelial dysfunction remains a difficult challenge. Endothelium insulin resistance and endoplasmic reticulum (ER) stress have been reported to contribute to endothelial dysfunction. Protein tyrosine phosphatase 1B (PTP1B) is a phosphatase bound to the ER known to regulate insulin sensitivity and ER stress. We hypothesize that diabetes alters PTP1B regulation of endothelial cell function and that inhibiting PTP1B may allow a strategy to prevent diabetes-induced endothelial dysfunction. Using immuno-histochemistry we showed that PTP1B is expressed in the endothelial cells of human saphenous vein, and in human primary aortic endothelial cells. Western-blot and quantitative real-time PCR analysis revealed that diabetes increased PTP1B and ER stress markers expression (CHOP: 1.4±0.2, GRP78: 1.5±0.2, XBP1spliced: 2.1±0.3, XBP1: 1.4±0.2 fold changes), in human saphenous veins. Deletion of PTP1B in obese diabetic mice (db/db) significantly reduced the level of expression (mRNA) of ER stress markers in mouse aorta. Incubation of mouse aortic rings with tunicamycin (tunica) or thapsigargin (thapsi), two ER stress inducers, increased the level of expression of ER stress markers (mRNA) and significantly reduced endothelium-dependent relaxation. (Tunica: -17±5%, thapsi: -30±6%, vs. DMSO). NO synthase inhibition with LNAME completely abolished endothelium-dependent relaxation in vehicle and tunica-treated rings. Neither the reactive oxygen species (ROS) scavenger tempol, nor catalase restored endothelial function, in tunica-treated rings. This suggests that ER stress induces a reduction in NO bioavailability independent of ROS. Aortic rings from PTP1B KO mice were protected against ER stress-induced endothelial dysfunction and exhibited a modest increase in ER stress markers compared to control rings, in response to tunica. All together these data demonstrate that PTP1B deletion prevents ER stress-induced endothelial dysfunction and that targeting PTP1B might be a beneficial approach to restore diabetic endothelial dysfunction.
Abstract 433: Role And Importance Of Antecedent Blood Pressure In Predicting Outcomes In Cardiovascular Disease
Authors: Bonifonte Anthony , Ayer Turgay , Veledar Emir , Clark Allison , Wilson Peter
Published: March 2022

Background:
Blood pressure levels have been shown to be predictive of subclinical and clinical cardiovascular disease (CVD).There is little published information concerning the influence of antecedent blood pressure levels and HTN on CVD risk. Our hypothesis was that adding antecedent HTN to predictive model will increase model quality and consequently our ability to tailor treatment to patients needs.

Methods:
We used data for 3344 participants from Framingham Heart Study Offspring who attended exam 3 and were followed up to 20 years for a first CVD event and analyzed prediction improvement reflected in changes in C-index, Net Reclassification Index (NRI) and the Integrated Discrimination Increment (IDI) values. Being exposed to serious critique in last 3 years, caused by improper overuse, this technique remains scientifically valid.

Results:

Antecedent BP is important independent predictor of CVD outcomes, surprisingly replacing current BP and BP treatment. Change in C-statistic was not statistically significant (p=0.19), which can be expected in saturated models with high c-index values. Both the IDI test (p=0.004) and the NRI test (p=0.046) indicate a significant improvement in the number of properly reclassified patients.

Discussion and conclusion:
Using information from personal medical history to properly diagnose patients is well established, while using that information in epidemiology was neglected, almost forbidden. Our hypothesis that adding antecedent BP to predictive model will help to properly classify patients was confirmed, so in reverse of roles, epidemiology result confirms practice, underlying need to use information from personal medical history.
Abstract 436: Over - Versus Under-Treatment of Older Hypertensives in Canada
Authors: Fodor George , Baker Penelope , Chen Li , Leenen Frans
Published: March 2022

In a re-analysis of data collected during the 2006 Ontario Survey on the Prevalence and Control of Hypertension (Leenen et al, CMAJ 2008), we focussed on the actual blood pressures in treated and untreated older and middle-aged hypertensives. In the older (60-79 years of age) population of 1,426,752 subjects, using traditional definitions the prevalence of hypertension was 49% compared to 21% in the middle-aged (40-59 years of age) population. Hypertension treatment and control rates were similarly high in middle-aged and older hypertensives at 67% and 64% respectively. 39% of older hypertensives were treated with a single antihypertensive drug, and of these 54% had a systolic BP level of < 130 mmHg, and 23% had a systolic BP < 120 mmHg. Of the 61% treated with combination therapy, 44% had a systolic BP < 120 mmHg. 13% of older hypertensives were untreated, and out of those approximately 90% had Stage 1 hypertension, with ~70% without additional risk factors.

Considering that monotherapy may lower systolic BP by < 10 mmHg, these findings suggest that there may be a major problem of over-diagnosis and/or over-treatment of hypertension in older adults overestimating substantially the actual prevalence of hypertension, creating an unnecessary burden on the health care system and exposing many older subjects to unnecessary risks of drug therapy.
Abstract 448: A CCR2 Anatagonist Reduces Vascular Fibrosis, Macrophage Accumulation and Blood Pressure in Angiotensin II-Treated Mice
Authors: Drummond Grant R , Moore Jeffrey P , Sakkal Samy , Krishnan Shalini M , Samuel Chrishan S , Diep Henry , Kemp-Harper Barbara K , Vinh Antony , Ricardo Sharon L , Guzik Tomasz J , Sobey Christopher G
Published: March 2022

Macrophages accumulate in the vascular wall during hypertension. However, their contribution to vascular remodeling remains to be established. In the present study we examined the polarization state of macrophages (M1/M2) in the aorta during experimental hypertension. We also investigated whether pharmacological antagonism of the chemokine receptors involved in macrophage accumulation reduces vascular remodeling and/or blood pressure (BP). Mice treated with angiotensin II (Ang II; 0.7 mg/kg/d) displayed elevated systolic BP (158±3 mmHg) compared with saline-treated animals (122±3 mmHg; P<0.001). Flow cytometry revealed that Ang II infusion caused a 2-fold increase in aortic macrophage (F4/80
+
) numbers (P<0.01), and that the majority of these cells expressed the M2 marker, CD206. A PCR array to identify chemokine receptor targets for pharmacological intervention revealed CCR2 and its ligand CCL8 to be upregulated by 2-fold and 4-fold, respectively in aortas from Ang II-treated mice (P<0.05). Taqman real-time PCR confirmed these findings and showed a significant positive correlation between CCR2 and CD206 expression in the vessel wall (P<0.001), suggesting that M2 macrophages were a source of this chemokine receptor. Intervention with a CCR2 antagonist (INCB3344, 30 mg/kg/d), commencing 7 d after initiation of Ang II infusion and continuing for a further 21 d, reduced macrophage accumulation in the aorta by ~60% (P<0.001). INCB334 completely prevented Ang II-induced increases in aortic collagen deposition and elastin breakdown (P<0.05), and tended to reduce vessel stiffening. Moreover, at the end of the treatment period, Ang II-infused mice that received INCB3344 had markedly lower systolic BP and heart weight to body weight ratios (137±4 mmHg and 5.5±0.1 mg/g, respectively) than did Ang II-infused animals that received vehicle (158±3 mmHg and 6.4±0.4 mg/g; P<0.05 for both parameters). Thus, CCR2 is crucial for M2 macrophage accumulation in the aorta during Ang II infusion in mice and may represent a promising target for future therapies aimed at reducing vascular fibrosis, cardiac hypertrophy and BP in hypertension.
Abstract 457: N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) Prevents The Development Of Nephritis And Skin Lesions In Mouse Model Of Systemic Lupus Erythematosus (MRL/lpr)
Authors: Liao Tang-Dong , Nakagawa Pablo , Janic Branislava , D’Ambrosio Martin , Worou Morel E , Yang Xiao-Ping , Rhaleb Nour-Eddine , Carretero Oscar A
Published: March 2022

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by development of auto antibodies against various self-antigens and deposition of immune complexes leading to inflammation, fibrosis and end organ damage. Ac-SDKP is a naturally occurring tetrapeptide that in hypertension and other diseases such as autoimmune myocarditis prevents inflammation and fibrosis in heart, kidney and vasculature. Here we tested the hypothesis that Ac-SDKP in a mouse model of SLE decreases autoimmune responses, thus preventing inflammation and renal damage. To test this, lupus mice were treated with Ac-SDKP for 20 weeks. Ac-SDKP treated lupus mice exhibited significant decrease in 1) macrophage and T cell infiltration, 2) expression of pro-inflammatory cytokines, 3) complement system activation and 4) expression of cell adhesion protein ICAM-1. In addition, the lupus mice had a significant improvement in renal function (GFR), decrease in glomerular damage and interstitial collagen deposition. Ac-SDKP also reduced the facial skin lesion. Ac-SDKP did not decrease the production of autoantibodies nor blood pressure. Therefore the protective effect of Ac-SDKP is most likely achieved by modulation of the activation of the complement system, expression of chemokines and ICAM-1, leading to a reduction of innate and adaptive immunity cell infiltration in the kidney
Abstract 459: Myeloid-Derived Suppressor Cells (MDSCs) Prevent Cyclosporine A-Induced Vascular and Renal Endothelial Cell Injury
Authors: Bounds Kelsey R , Chiasson Valorie L , Roman Marcos H , Mitchell Brett M
Published: March 2022

Cyclosporine A (CsA) is an immunosuppressive drug that can cause vascular and renal endothelial dysfunction and hypertension. Myeloid-derived suppressor cells (MDSCs) exert anti-inflammatory activity by inhibiting various innate and adaptive immune cells; however, the direct effects of CsA on MDSCs and whether MDSCs can prevent the detrimental CsA effects on endothelial cells are unknown. We hypothesized that CsA directly decreases MDSCs and that MDSCs prevent CsA-induced endothelial injury. CsA (50 uM) treatment of CD11b+/Gr-1+ MDSCs isolated from male C57BL6/J mice for 24 hours caused a dose-dependent decrease in MDSCs (Total CD11b+/Gr-1+ cell number: Veh=6,358,000, CsA 10 uM=3,505,000, CsA 25 uM=2,685,000, CsA 50 uM=1,425,000). Mouse cardiac vascular and glomerular endothelial cells were treated with CsA (50 uM) in the absence and presence of 75,000 isolated MDSCs for 24 hours. CsA significantly increased fibronectin and ICAM protein levels 1.5-2-fold in both vascular and glomerular endothelial cells compared to vehicle-treated endothelial cells. The presence of MDSCs prevented the CsA-induced increase in fibronectin and ICAM while having no effects in vehicle-treated endothelial cells. Additionally, CsA (50 uM) treatment of aortas isolated from male C57BL6/J mice for 24 hours markedly decreased maximal acetylcholine-induced relaxation responses and this was partially restored by co-incubation with MDSCs, and isolated kidneys treated with CsA had increased levels of fibronectin and this was ameliorated by co-incubation with MDSCs. In conclusion, CsA-induced decreases in MDSCs may play a role in endothelial cell injury and the augmentation of MDSCs may prevent the vascular and renal dysfunction and hypertension caused by CsA.
Abstract 463: Dietary Intake and Arterial Compliance in a Cohort of People with Type 1 and Type 2 Diabetes
Authors: Petersen Kristina S , Clifton Peter M , Keogh Jennifer B
Published: March 2022

Background:
Diabetes is associated with a higher risk of cardiovascular disease (CVD) and diet is a modifiable risk factor. However, few studies have examined the association between dietary intake and measures of arterial compliance, a well-established predictor of CVD, in people with diabetes. The aim was to investigate the relationship between dietary intake, augmentation index (AI), and pulse wave velocity (PWV) in a cohort of people with type 1 and type 2 diabetes.

Methods:
Participants were adults with type 1 or type 2 diabetes who completed the Dietary Questionnaire for Epidemiological Studies (v2) Food Frequency Questionnaire and had carotid- femoral PWV (n=95) and/or AI (n=110) measured using a SphygmoCor
®
XCEL (Sydney, Australia). A random spot urine sample was provided and predicted 24 hour urinary sodium excretion was calculated using the Kawasaki equation.

Results:
A negative linear relationship was observed between tertile of dairy intake and PWV which persisted after multivariate adjustment (demographic variables and cardiovascular risk factors) (t=-2.4; p=0.019). PWV was 0.9m/s (95% CI 0.2 to 1.7m/s) lower in the highest tertile (>434g/d) compared to the lowest tertile (<260g/d) of dairy consumption (p=0.02 after multivariate adjustment). There was also a negative linear relationship between AI and tertile of fibre intake (t=-2.6; p=0.012) and tertile of potassium intake (t=-2.4; p=0.019), after multivariate adjustment. Predicted 24hr sodium excretion was positively associated with AI (t=2.2; p=0.029) after adjustment. The lowest tertile of sodium excretion (<69mmol/d) was associated with a 4.7% lower AI compared with the high tertile (>90mmol/d) (p=0.02 after multivariate adjustment). It was estimated that a 100mmol increase in sodium excretion would result in an 8.2% increase in AI.

Conclusion:
A dietary pattern that is high in dairy, fibre, and potassium and lower in sodium may improve arterial compliance in people with diabetes.
Abstract 471: Oral Formulation Of Angiotensin-(1-7) Improves Metabolic Parameters And Insulin-signaling Pathway In Adipose Tissue Of Rats With Metabolic Syndrome
Authors: Santos Robaon A , de Castro Uberdan G , Guerra Renata , Barbosa Maria A , Queiroz Kaina B , Nicolato Roney Luiz C , Silva Marcelo Eustáquio , Campagnole-Santos Maria J , Alzamora Andreia C
Published: March 2022

In this study we evaluated the potential therapeutic effect of the oral formulation of angiotensin-(1-7) [Ang-(1-7)] included in cyclodextrin (HPβCD) on biochemical parameters, expression of adipokines and insulin-signaling pathway in the retroperitoneal adipose tissue (rWAT) of rats with metabolic syndrome (MS) induced by high fat diet. Rats were subjected to AIN-93 (CT) or high-fat (MS) diet for 13 weeks. HβCD-Ang-(1-7) or vehicle (V) was given in the last 6 weeks by gavage. MS-V rats presented increased body weight (314±10 vs 283±6 g, CT), adipositivity index (8±0.3 vs 5±0.1, CT), fasting glucose plasma levels (7±0.1 vs 6±0.1 mmol/L, CT), total cholesterol (2.0±0.1 vs 1.7±0.03 mmol/L, CT) and increased alanine aminotransferase (ALT; 65±2 vs 57±2 a.u., CT). In addition, there was an increase in mRNA levels (a.u.) of adipokines, leptin (18±6 vs 5±3), resistin (10±3 vs 3±0.9), adipisina (35±10 vs 13±3), TNF-α (0.05±0.01 vs 0.01±0.002) and COX-2 associated to a decrease in the mRNA expression of adiponectin (34±5 vs 89±11), insulin receptor (Irs; 0.3±0.04 vs 0.8±0.1), Irs-1 (0.5±0.1 vs 1.4±0.04), Irs-2 (0.2±0.03 vs 0.6±0.1), Akt-2 (2±0.6 vs 4±0,3) and GLUT-4 (0.2±0.03 vs 0.5±0.01) in the rWAT of MS rats. However, MS rats treated with HβCD-Ang-(1-7) presented reduction in body weight, adiposity index (7 ± 03), in fasting glucose plasma levels (7±0.1 mmol/L), total cholesterol and ALT mRNA (58±2 a.u.) compared to MS untreated rats. HβCD-Ang-(1-7) treatment also induced a decrease in the mRNA expression (a.u.) of leptin (6±11), resistin (2±0.4), adipisina (9±1), TNF-α (0.008±0.0006) and COX-2 and increase in Irs-1(0.73±0.1), Irs-2 (0.3±0.09) and GLUT-4 (0.34±0.03) in rWAT compared to MS-V rats. Our data showed that treatment with HβCD-Ang-(1-7) reversed the changes in metabolic and biometrics parameters, reduced the expression of pro-inflammatory adipokines and improved the insulin-signaling pathway in the rWAT of rats with MS induced by high-fat diet.
Abstract 501: Up-Regulation of Inflammatory Mediators and Markers of Metabolic Deraignment in ESRD as Studied by Biochip Array Analysis
Authors: Bansal Vinod , Syed Daneyal , Hoppensteadt Debra , Fareed Jawed
Published: March 2022

Introduction:
End stage renal disease represents a complex syndrome with multiple pathophysiological processes involving vascular, inflammatory, thrombotic, and metabolic deraignment. To understand the pathophysiology of this complex syndrome, profiling of mediators of inflammation and metabolic deraignment provides a unique approach. This study was designed to utilize biochip array technology to compare the biomarker profiles of a maintenance hemodialysis cohort (n=81) with healthy normal male and female volunteers (n=41).

Materials and Methods:
Blood samples from the ESRD group were collected prior to the hemodialysis, centrifuged immediately, and frozen at -80 degrees Celsius. The normal samples represented citrated plasmas from 41 healthy individuals, which were processed in an identical manner as the ESRD group. High sensitivity inflammatory cytokine chips to profile IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, IFN-gamma, IL1-alpha, IL1-beta, MCP1, and EGF and metabolic array chip to analyze C-Peptide, ferritin, resistin, insulin, leptin, and PAI-1 were used employing Evidence Investigator (Randox, UK).

Results:
In the inflammatory biochip array analysis, except for IL-2 and EGF, all the inflammatory biomarkers were found to be significantly higher than the normal group (p<0.0001). The ESRD group exhibited marked variations in the circulating levels of the inflammatory mediators and the extent of increase also varied. In the metabolic chip array analysis, all of the markers of metabolic deraignment were significantly increased, except insulin, where a trend towards increased levels was noted.

Conclusions:
These results clearly demonstrate the complexity of the pathophysiologic event in ESRD patients. A widespread increase in the inflammatory mediators, coupled with the higher levels of metabolic deraignment markers suggest that the ESRD patients not only have an ongoing inflammatory process but also sustain metabolic deraignment as evident by the results obtained by the metabolic array chip. Profiling of these mediators not only provides an understanding of the pathogenesis of this condition, but may be helpful in the risk stratification and clinical management of these patients.
Abstract 509: Thermodynamic Energy Stability and In Vitro Potassium Exchange by ZS-9, a Novel Selective Cation Trap for Treatment of Hyperkalemia
Authors: Singh Bhupinder , Yang Alex , Nuttall Mark , Low John , Rasmussen Henrik S
Published: March 2022

Hyperkalemia is associated with significant mortality and often leads to dose reduction or cessation of RAAS inhibitors, a drug class with proven cardio- and reno-protective effects. The only approved hyperkalemia therapy in the US is a nonselective organic resin sodium polystyrene sulfonate (SPS), which is associated with Na
+
loading, potentially worsening fluid retention, and hypertension. ZS-9 is an insoluble, nonsystemic inorganic cation exchanger that preferentially entraps K
+
over other monovalent (eg, Na
+
) or divalent (Ca
2+
, Mg
2+
) cations in the GI tract. ZS-9 has a lattice structure that provides high K
+
selectivity, analogous to the physiologic selectivity filters in in vivo K
+
channels. Here we compared in vitro K
+
exchange capacity (KEC) for ZS-9 and SPS and performed thermodynamic stability modeling for various cations (K
+
, Na
+
, Ca
2+
, Mg
2+
) within the ZS-9 lattice structure.

For KEC assessments, ZS-9 or SPS were added to a solution containing K
+
, Ca
2+
and Mg
2+
. After equilibration, binding of each ion was quantified. Selectivity ratios were calculated as [K
+
]/([Ca
2+
]+[Mg
2+
]). The predicted energies for different cation forms of ZS-9 (ie, Na-ZS-9, K-ZS-9, Ca-ZS-9 and Mg-ZS-9), in addition to alkali and alkaline earth oxides from models, was used to estimate cation exchange energies in ZS-9. All energies were computed relative to the Na
+
form of ZS-9 (“reference state”).

ZS-9 with K
+
was calculated to be more thermodynamically stable than ZS-9 with Na
+
, Ca
2+
, or Mg
2+
, with the K
+
form of ZS-9 estimated to be 20 kcal/mol more stable than the Na+ form. Preference for K
+
was confirmed in vitro. Exchange capacities of ZS-9 for Ca
2+
or Mg
2+
were below the LLOQ (0.05 mEq/g). At a 1:1:1 ratio mixture of K
+
:Ca
2+
:Mg
2+
, KEC was 0.3 mEq/g for SPS vs. 2.7 mEq/g for ZS-9, indicating that ZS-9 has ≥25-fold increased selectivity for K
+
vs Ca
2+
or Mg
2+
. Selectivity of SPS for K
+
was 0.2-fold that of the divalent cations.

In the presence of K
+
, Ca
2+
, and Mg
2+
, ZS-9 appeared to be highly selective for K
+
, whereas SPS was more selective for Ca
2+
, consistent with earlier reports. In addition, ZS-9’s capacity for K
+
was ~9-fold that of SPS. If approved, ZS-9 may represent a novel first-in-class treatment for hyperkalemia with improved selectivity and capacity for K
+
.
Abstract 516: Intestinal Cell Gastrin Secretion is Regulated by a Sodium-Induced Increase in PPAR-A that is Mediated by Dopaminergic Stimulation
Authors: Gildea John J , Zhang Chi , Bigler Wang Dora , Tran Hanh T , Felder Robin A
Published: March 2022

Recently a novel gastro-renal axis has been discovered for the regulation of renal sodium excretion at the moment of sodium ingestion. Cells in the digestive track secrete gastrin in response to sodium ingestion which stimulates an increase in renal sodium excretion. Dopamine is largely responsible for the regulation of sodium excretion in the kidney, and has also been shown to decrease sodium transport in intestines. We hypothesized that dopamine might also be involved with the regulation of gastrin secretion in a human intestinal gastrin secreting cell line (SW626, ATCC, Manassas VA) that also contains dopaminergic receptors. We used the ionophore monensin (MON) to increase intracellular sodium in SW626 cells in order to simulate an oral sodium load. We also stimulated the cells with the dopaminergic agonist SKF38393 (SKF, D
1
-like receptor agonist). These two manipulations (compared to vehicle VEH) caused increased relative gene expression of gastrin (ΔΔCt gastrin to actin, VEH 0.00638±0.00283; MON 0.3913372±0.0985926; SKF 0.2139286±0.0253672; P<0.01 VEH vs MON or SKF, N=6 per group). Peroxisome proliferator-activated receptor-alpha (PPAR-α) associates with the gastrin transcriptional promoter by CHIP-SEQ (ENCODE database) and PPAR-α activation by fenofibrate (FENO) has been shown to reduce blood pressure and increase sodium excretion. We hypothesized that PPAR alpha would increase gastrin expression. FENO increased the immunoreactive gastrin and this effect was blocked by a PPAR alpha antagonist GW6471 (VEH 16,440±2,438 RFU; FENO 42,639±4,891; FENO+GW 19,367±1,865; P<0.001 VEH vs FENO and FENO vs FENO+GW). Stimulation with MON or SKF also significantly stimulated the production of immunoreactive gastrin (VEH 16,440±2,438 RFU; MON 22,345±935; SKF 24,386±1,124; GW alone 10,235±1,214; P<0.05 VEH vs MON, SKF, or GW, N=6). SW626 appear to secrete gastrin, therefore we tested the ability of secreted gastrin obtained from the media bathing SW626 cells to bind to i22 human renal proximal tubule cells (RPTC). After incubation of i22 RPTC in SW626 media for 24 hours, we detected surface gastrin staining on the i22 RPTC. We conclude that gastrin is produced by SW626 cells, is secreted into the growth media, and can bind to cell surface receptors on RPTC.
Abstract 523: Diabetes Promotes the Development of Aortic Aneurysm in Mice with Hypertension
Authors: Wongjarupong Asarn , Knudsen Bruce E , Hartono Stella , Grande Joseph P
Published: March 2022

Introduction:
The estimated prevalence of diabetes mellitus (DM) is 8.4% among adults in the U.S. Cardiovascular disease is a major complication of DM. We have observed that diabetic mice, but not non-diabetic mice, undergoing 2kidney-1clip (2K1C) surgery inducing renovascular hypertension had high mortality attributed to ruptured aortic aneurysm. However, the effect of diabetes on formation of aortic aneurysm remains unclear.

Method:
In our previous experiment, 163 mice were divided into 4 groups: leptin KO 2K1C (db 2K1C, n=52), leptin KO Sham (db Sham, n=20), wild type 2K1C (dbc 2K1C, n=62), and wild type Sham (dbc Sham, n=29), respectively. We found that 42% (22/52) of db 2K1C died, from ruptured aortic aneurysms examined by gross examination, within 120 days, while others survived 100%. In this study, available H&E aortic sections of survived mice (n=74): db 2K1C (n=30), db Sham (n=15), dbc 2K1C (n=20), and dbc Sham (n=9) were examined 2K1C surgery in a blinded fashion. Aortic lesions defined as necrosis, thrombosis and dropped-out smooth muscle cells, which are signs of early formation of aortic aneurysm, were identified. Inflammation was graded as mild, moderate and severe inflammation.

Result:
The db 2K1C mice had a significantly greater aortic lesions (16/30, 53%) compared to the db Sham (13/15, 20%); and dbc 2K1C mice (1/20, 5%), p=0.002 and 0.0001, respectively. The number of aortic lesions in the dbc 2K1C group was not statistically different from that of the dbc Sham group (p=0.5). Similarly, the number of aortic lesions in the db Sham group was not statistically different from that of the dbc Sham group (0/9, 0%), p=0.125. Moderate-to-severe inflammation in the tunica media was more prevalence in db mice than the dbc mice.

Conclusion:
We report for the first time an increased incidence of aortic lesions in db 2K1C compared to dbc and sham groups. This finding suggests that diabetes contributes to aortic aneurysm formation in mice with renal artery stenosis, potentially due to induction of inflammation. Further studies to confirm a mechanism are warranted.
Abstract 529: Treatment With Telmisartan Inhibits Kidney Injury in High Salt-loading Tsukuba Hypertensive Mice With Attenuation of Sympathetic Nervous System
Authors: Iwanami Jun , Mogi Masaki , Tsukuda Kana , Wang Xiao-Li , Kukita Masanori , Nakaoka Hirotomo , Chisaka Toshiyuki , Bai Hui-Yu , Shang Bao-Shao , Horiuchi Masatsugu
Published: March 2022

Objective:
It is well known that excessive salt intake and/or activation of renin-angiotensin system (RAS) increases blood pressure in part via an increase in sympathetic nerve activity. To further examine the interaction of these factors, we employed transgenic mice carrying both the human renin and angiotensinogen genes (Tsukuba hypertensive mice) and studied the effects of high salt loading and telmisartan on mortality, blood pressure and kidney functions.

Design and Method:
Ten-week-old male Tsukuba hypertensive mice (TH) were administrated control chow or the chow containing 8% NaCl with or without telmisartan (1 mg/kg/day) and PPAR-γ antagonist, GW9662, in drinking water for 8 weeks. Blood pressure was assessed by radio telemetry method. Urine samples were obtained before and after salt-loading. Concentrations of urinary adrenalin and noradrenalin were measured by ELISA. Expressions of ENaC-α, P47 and p67-phox mRNA in the kidney were measured by RT-PCR.

Results:
Survival rate 8 weeks after treatment was decreased in salt-loading TH (sTH) mice compared with control TH (cTH). This decrease was improved by the treatment with telmisartan. Blood pressure in sTH mice was significantly higher compared with cTH mice and the treatment with telmisartan mice decreased blood pressure. Urinary sodium concentration was higher in sTH mice compared with cTH mice and treatment with telmisartan further increased urinary sodium concentration in sTH mice. Expression of ENaC-α in did not between sTH mice and cTH mice. Treatment with telmisartan decreased expression of ENaC-α. Urinary adrenalin and noradrenalin concentrations were higher in sTH mice compared with cTH mice. These increases were attenuated by treatment with telmisartan. Expressions of p47 and p67-phox in sTH mice were increased more markedly compared with cTH mice. These increase were also attenuated by treatment with telmisartan. However, these possible protective effects of telmisartan were not influenced by the co-treatment with GW9662.

Conclusion:
These results suggested that salt-loading enhanced an increase in blood pressure, superoxide anion production, sympathetic activity and mortality in concert with activated RAS.
Abstract 535: Chronic High Sodium Diet Attenuates The Development Of Renal Injury And Inflammation In Murine Systemic Lupus Erythematosus
Authors: Broome Hanna J , Ryan Michael J
Published: March 2022

Systemic lupus erythematosus (SLE) is an autoimmune disorder with prevalent renal disease, a consequence of increased and abnormal survival of B and T lymphocytes. Recent evidence suggests that dietary salt may be an important environmental factor that promotes certain autoimmune diseases by stimulating pro-inflammatory T lymphocyte differentiation. Therefore, we hypothesized that a long-term high salt diet would accelerate the progression of renal disease during SLE. In order to test this, an established experimental model of SLE (female NZBWF1 mice) was fed a standard (0.4% NaCl) or high salt (4% NaCl) diet starting at 10 weeks of age. Sodium intake (in meq/day) was measured three times throughout the study and was significantly greater in high salt fed animals at 16 weeks of age (2.9±0.6 vs. 0.5±0.06 , p=0.01), 23 weeks (4.8±0.8 vs. 0.6±0.06, p=0.004) and 29 weeks (5.7±0.6 vs. 0.6±0.05, p=0.0007). Urinary albumin was monitored monthly until 30 weeks then weekly by dipstick assay as a marker of renal injury until 34 weeks of age, at which time catheters were implanted into the carotid arteries for measurement of mean arterial pressure (MAP). Mice were then euthanized and kidneys collected. Renal cortex enriched mRNA was analyzed by qRT-PCR for inflammatory markers, and renal medullary enriched mRNA was analyzed for expression of serum glucocorticoid kinase 1 (SGK1), a salt-sensing kinase. Counter to our hypothesis, 75% (9/12) of animals on a normal diet developed albuminuria (≥ 100 mg/dL), whereas 17% (2/12) of the high salt fed animals developed albuminuria (p=0.0002). MAP was not different between groups (132±4 mmHg 0.4% NaCl vs. 144±8 mmHg 4% NaCl), nor was renal medullary expression of SGK1 mRNA (0.25±0.03 vs. 0.20±0.04). However, renal cortical interleukin-2 (IL-2), a cytokine important for T cell differentiation, expression was significantly lower in high salt fed animals (0.31±0.03 vs. 1.16±0.34, p=0.03). These data suggest that a long-term high salt diet may protect against the renal injury associated with SLE, possibly through alterations in IL-2 mediated T cell differentiation, without a significant effect on MAP. Furthermore, the data advance our overall understanding of how dietary salt may impact different autoimmune disorders.
Abstract 537: Rgulation of ALG-2 in NAADP-induced Ca2+ Release via TRP-ML1 Channel in Mouse Coronary Arterial Myocytes
Authors: Xu Ming , Li Xiao-Xue , Xia Min , Zhang Yang , Li Pin-Lan
Published: March 2022

We have demonstrated that nicotinic acid adenine dinucleotide phosphate (NAADP) sensitive Ca2+ release channel is characteristic of lysosomal TRP-ML1 in coronary arterial myocytes (CAMs). The present study hypothesized that NAADP induces Ca2+ release from TRP-ML1 channels via apoptosis-linked gene 2 (ALG-2), which was reported to act as a Ca2+ sensor that modulates the function of TRP-ML1. By RT-PCR and Western blot analysis, we demonstrated that ALG-2 is abundantly expressed in the cytosol of mouse CAMs. Confocal microscopy found that FasL and ET-1, well-known NAADP stimulators, increased the colocalization coefficient of ALG-2-GFP with TRP-ML1-RFP in CAMs from 0.17±0.033 to 0.3±0.044 and 0.3±0.09, respectively. In addition, the TRP-ML1 level was significantly increased in FasL or ET-1-treated CAMs after immunoprecipitation with anti-ALG-2, confirming that the interaction of ALG-2 with TRP-ML1 was enhanced in response to NAADP production. Furthermore, NAADP competitively inhibited recombinant ALG-2 protein binding with tis-NED-19, which fluorescently labels NAADP receptors. Importantly, fluorescent microscopic image analysis showed that NAADP-induced Ca2+ release was reduced from 380±42.03 nM to 216.7±14.7 nM after the transfection with ALG-2 siRNA in CAMs, which was also found after the treatment with FasL or ET-1. Our findings support the view that ALG-2 can directly bind with NAADP, which mediates the action of NAADP through lysosomal TRP-ML1 channels. Such results throw light on NAADP-TRP-ML1 working model in vascular cells.
Abstract 553: High-Intensity Interval Training Partly Restores Thrombotic Microangiopathy in an Experimental Model of Hypertensive Renal Injury
Authors: Silva Gustavo J , Langlo Knut A , Rolim Natale , Moreira Jose B , Medeiros Alessandra , Alves Marcia , Yang Xiaojuan , Gusky Till , Lesko Paulina , Bækkerud Fredrik H , Ness Henning O , Aasarød Knut , Hallan Stein I , Wisløff Ulrik
Published: March 2022

To investigate whether high intensity interval training can reduce hypertensive renal injury in Dahl salt-sensitive rats. Adult females Dahl salt-sensitivity (Dahl SS) rats were randomized in three groups: sedentary low-salt diet (LS, N=20: 0.3% NaCl), sedentary high-salt diet (HS, N=40: 8% NaCl) or high-salt diet submitted to high intensity interval training (HIT, N=20: 8% NaCl + HIT, 3x38 min/wk; 60% HRmax adding 4 intervals of 4 min at 90% HRmax). After 30 weeks of follow-up, 24h urine has been collected in metabolic cages, followed by tissue harvesting. HIT did not affect the development of hypertension induced by high-salt intake in the Dahl SS rats (147±55 vs 209±80* vs 203±79* mm Hg, for LS, HS and HIT, respectively; *p<0,05 compared to LS). However, total mortality was significantly reduced (p=0.0022) in the HIT group (8/20 events) compared to HS (23/40 events) and LS (1/20 events) groups (94.7% vs 39.9% vs 59% survival, for LS, HS and HIT respectively), most of the deaths caused by stroke (69.6% incidence in HS rats). Chronic high blood pressure caused microalbuminuria that was not reverted by exercise (0.65±0.34 vs 95±42* vs 72±37* mg/24h, for LS, HS and HIT, respectively; *p<0.05 compared to LH). Creatinine clearance was not changed by either hypertension or exercise (1.91±0.58 vs 1.73±0.85 vs 1.60±0.66 ml/min, for LS, HS and HIT, respectively; p=0.53). Histological analysis of the renal injury reveled that HIT did not reverse the expansion of mesangial matrix (125±40 vs 179±44* vs 188±47*, for LS, HS and HIT, respectively; *p<0.05 compared to LS) and no glomerulosclerosis were detected (69±36 vs 97±36 vs 117±57, for LS, HS and HIT, respectively; p=0.12). Interesting, HIT significantly reduced (7/10) the number of nephrons with thrombotic microangiopathy observed in the HS (10/11) compared to LS (0/15) group (0.00±0.00 vs 8.82±2.82* vs 6.60±3.06 counts/kidney, for LS, HS and HIT, respectively; *p<0.05 compared to LS). Taken together our data demonstrate that, although no changes have been observed in the hypertension status and microalbuminuria, HIT partially reverted the high thrombotic microangiopathy incidence in the Dahl salt model, and may explain the reduction in the mortality caused by stroke.
Abstract 561: Melanocortin-4 Receptor Signaling Is Required For The Anorexic, Cardiovascular And Antidiabetic Actions Of Leptin In Diabetic Rats.
Authors: da Silva Alexandre A , do Carmo Jussara M , Hall John E
Published: March 2022

We previously demonstrated that leptin has powerful antidiabetic actions that are mediated mainly through central nervous system (CNS) effects. In this study we tested the specific importance of melanocortin-4 receptor (MC4R) in mediating leptin’s ability to suppress food intake, to increase blood pressure (BP) and heart rate (HR), and to normalize glucose levels in insulin-dependent diabetes. MC4R knockout (MC4R
-/-
, n=5) and control wild-type rats (WT, n=4) were implanted with BP telemetry transmitters and an intracerebroventricular (ICV) cannula was inserted into the brain lateral ventricle. After 10 days of recovery, BP and HR were measured 24-hrs/day. After stable baseline measurements for 5 days, a single bolus injection of streptozotocin (STZ) (50 mg/kg, ip) was used to induce insulin-dependent diabetes. Eight days after STZ injection, an osmotic pump was implanted subcutaneously and connected to the ICV cannula to deliver leptin (15 μg/day) for 7 days. At baseline, MC4R
-/-
rats ate 20% more and were 40% heavier than WT rats. Despite being markedly obese, BP was similar (112±2 vs. 111±2 mmHg) and HR was lower in MC4R
-/-
rats (320±6 vs. 347±5 bpm). Induction of diabetes increased food intake (30%) and reduced BP (~17 mmHg) and HR (~61 bpm) in WT rats, while food intake, BP and HR were reduced by ~10%, 7 mmHg and 33 bpm in MC4R
-/-
rats. Leptin treatment for 7 days normalized glucose levels (437±10 to 136±18 mg/dl), reduced food intake (40%), and increased HR (+60 bpm) and BP (+9 mmHg) in WT rats. However, only modest changes in glucose levels (367±16 to 326±23 mg/dl), food intake (5%), HR (+16 bpm) and BP (+4 mmHg) were observed in MC4R
-/-
rats. These results indicate that intact CNS MC4R signaling is necessary for leptin to exert its chronic anorexic, cardiovascular and antidiabetic actions.
Abstract 564: Reduction of Blood Pressure with Antisense Oligodeoxynucleotides Against Chemerin
Authors: Mahon Bridget , Mullick Adam , Burnett Robert , Watts Stephanie W
Published: March 2022

Chemerin promotes dendritic cells and macrophage migration to sites of inflammation. The liver and adipose tissues are sources of chemerin. There is a direct relationship of BMI, plasma chemerin concentration and blood pressure. Chemerin causes arterial contraction that is amplified in tissues with endothelial dysfunction. We hypothesized that if chemerin supported hypertension through such mechanisms, removal of chemerin should decrease blood pressure. Ten antisense oligodeoxynucleotides (ASOs) were developed against the retinoic acid receptor responder protein 2 (RARRES2, chemerin gene). ASO642647 was chosen as a lead based on excellent tolerance and minimal changes in a standard clinical panel. The model chosen was the deoxycorticosterone acetate (DOCA) salt hypertensive rat as this model shows profound endothelial dysfunction. Rats were uninephrectomized, given DOCA (s.c., 150 mg/kg) and placed on 1.0% NaCl + 0.2 % KCl. One week later, radiotelemeters were implanted for measurement of systolic, diastolic, mean arterial blood pressure and heart rate. ASOs (control nonsense or chemerin) were injected ip weekly over 3.5 weeks (25 mg/kg week 1 and 2; 12.5 mg/kg week 3 and 4) when DOCA-salt hypertension was stable (week 3.5). A group of DOCA-salt rats also received injections of PBS (ASO vehicle). Two days after the 4th injection, tissues were dissected for measurement of chemerin mRNA and blood taken for measures of catecholamines. ASO Chemerin reduced mean arterial blood pressure vs ASO control and PBS group within the first week, and remained reduced by ~17+/- 2 mm Hg at the time of sacrifice. Target organ mass (liver, heart and kidneys) was not different between ASO control and ASO chemerin group. HPLC revealed that circulating norepinephrine (NE) and dopamine (DA) were reduced 50% in the ASO chemerin vs control or PBS group. Real time RT-PCR validated knockdown of chemerin in ASO chemerin treated animals, with reductions as follows (vs ASO control, +/-3%): Liver (94%), epididymal fat (90%), retroperitoneal fat (90% ) and mesenteric perivascular adipose tissue (85%). These experiments provide powerful support for chemerin playing a role in blood pressure elevation.
Abstract 566: A Genetic Locus on Mouse Chromosome 11 controls Endothelial Dysfunction
Authors: Smolock Elaine M , Burke Ryan M , Wang Chenjing , Thomas Tamlyn , Batchu Sri N , Zettel Martha , Fujiwara Keigi , Berk Bradford C , Korshunov Vyacheslav A
Published: March 2022

Introduction:
Carotid intima formation is a significant risk factor for cardiovascular disease. C3H/FeJ (C3H/F) and SJL/J (SJL) inbred mouse strains differ in susceptibility to immune and vascular traits. Using a congenic approach we demonstrated that the Intima modifier 2 (
Im
2) locus on chromosome 11 regulates leukocyte infiltration. We sought to determine whether inflammation was due to changes in circulating immune cells or activation of vascular wall cells in genetically pure
Im
2 (C3H/F.SJL.11.1) mice.

Methods and Results:
Complete blood counts showed no differences in circulating cells between C3H/F and C3H/F.SJL.11.1 compared to SJL mice. Aortic vascular cell adhesion molecule-1 (VCAM-1) total protein levels were dramatically increased in SJL and C3H/F.SJL.11.1 compared to C3H/F mice. Immunostaining of aortic endothelial cells (EC) showed a significant increase in VCAM-1 expression area (um
2
) in SJL (0.18±0.02) and C3H/F.SJL.11.1 (0.19±0.02) compared to C3H/F (0.09±0.02) under steady flow conditions. In regions of disturbed flow we observed a significant decrease in EC area, measured by membrane staining, in SJL (500±50) and C3H/F.SJL.11.1 (500±30) versus C3H/F (800±50). Vascular permeability measured by the Miles assay (Evans’ blue dye ug per mg of aorta) was significantly higher in SJL (0.043±0.006) compared to C3H/F (0.010±0.005). In C3H/F.SJL.11.1 congenic mice vascular permeability varied from 0.006 to 0.665 but the average trended toward SJL values.

Conclusions:
Our results indicate that
Im
2 regulation of leukocyte infiltration is mediated by EC inflammation and permeability. Comparative analysis of gene polymorphisms in human regions syntenic with the mouse
Im
2 locus provides new insights into candidate genes that regulate vascular wall inflammation.
Abstract 571: Osteogenic Factors Regulate Vascular Function Through Reactive Oxygen Species And Toll-Like Receptor 4 Activation: Role In Hypertension.
Authors: Neves Karla B , Lopes Rheure A , Burger Dylan E , Graham Delyth , Touyz Rhian M , Montezano Augusto C
Published: March 2022

Osteogenic factors (OF), such as osteoprotegerin (OPG) and fetuin-A (FTA), play a role in regulating vascular calcification. Plasma levels of OPG and FTA are elevated in metabolic diseases and correlate with endothelial dysfunction, a hallmark of hypertension. Molecular mechanisms for this are elusive and the association with hypertension is unclear. Here, we postulated that OFs influence vascular function. Mesenteric arteries and VSMCs from WKY and SHRSP rats were studied. Vascular function was analyzed by wire myography. ROS production was assessed by chemiluminescence. Protein expression was analyzed by immunoblotting. Calcification was assessed by Von Kossa staining. WKY vessels exposed to OPG and FTA (50 ng/mL-30 min) were less sensitive to acetylcholine (Ach)-induced relaxation; an effect blocked by N-acetylcysteine (NAC-antioxidant), tempol (SOD mimetic) and ML171 (Nox1 inhibitor). Sensitivity to phenylephrine was increased by OPG and FTA, an effect mediated by ROS and Nox1. FTA has been characterized as an endogenous ligand to toll-like receptor 4 (TLR4). Inhibition of TLR4 with CLI095 blocked effects of FTA in WKY vessels. Stimulation with OPG (100ng/5 min) and FTA (50ng/5 min) increased ROS production (≈50%) in VSMCs from WKY rats (p<0.05 vs vehicle). ROS generation induced by OPG and FTA was blocked by GKT137831 (Nox1/Nox4 inhibitor) and PP2 (c-Src inhibitor). MAPK activation was also induced by OPG (JNK: 75%, p38: 110%, ERK1/2: 100%; p<0.05 vs vehicle), but FTA only increased ERK1/2 activation (50%, p<0.05 vs vehicle) in VSMCs from WKY rats. In VSMCs from SHRSP rats, basal levels of OPG (20%) and osterix (86%) were increased. In addition, aorta from SHRSP, but not WKY, exhibited positive Von Kossa staining. Levels of OPG (75%), FTA (40%) and osterix (50%) protein levels were increased in LinA3 mice, a mouse model which has chronically elevated Ang II levels (p<0.05 vs WT). In conclusion, our data suggest that in hypertension, OPG and FTA may influence vascular function through redox-sensitive TLR4-dependent signalling. Identification of vascular calcification-derived osteogenic factors as modulators of VSMCs biology provides new insights into molecular mechanisms of vascular dysfunction in hypertension.