bluuebunny/crossref_metadata_2025_split huggingface dataset:
PS-BPB04-5: SELECTIVE ACTIVATION OF GLUTAMATERGIC NEURONS WITHIN THE SFO EVOKES A PRESSOR RESPONSE THAT IS DEPENDENT ON THE PVN
Authors: Acquoy Monique L Van , Nedelkoska Teodora , Burke Peter G R , Hildreth Cara M 
Published: January 2023

Objective:
An overactive glutamatergic projection from the subfornical organ (SFO) to the paraventricular nucleus (PVN) is associated with the development of hypertension. Despite this, the functional relationship of this pathway under normotensive conditions remains poorly defined within the literature. Without this knowledge, it remains unknown if a dysfunction of this pathway is the underlying cause of hypertension or rather simply a consequence of the underlying hypertensive disease pathology. As such, the aim of this study is to identify the short-term function of the SFO and PVN pathway under normotensive conditions.

Design and Methods:
An optogenetic viral vector (pAAV-CaMKIIa-hChR2 (H134R)-EYFP) was microinjected into the SFO of Lewis rats (n = 5). Three weeks later, rats were anaesthetised and instrumented to record blood pressure and sympathetic nerve activity, administer intravenous drugs and to photo-stimulate glutamatergic SFO neurons.

Results:
Photostimulation of glutamatergic SFO neurons produced a pressor response that was abolished by inhibition of neuronal transmission within the PVN (9 ± 1mmHg vs 3 ± 1mmHg peak change from baseline, baseline vs PVN inhibition, n = 2 rats, 3 replicates per rat). In a separate cohort (n = 3 rats, 5 replicates per rat), we determined if the pressor response elicited was dependant on the sympathetic nervous system and/or the release of the vasoactive hormone vasopressin. Blockade of sympathetic vasomotor outflow attenuated the pressor response by 76%, whereas combined inhibition of sympathetic outflow and vasopressin 1a receptors attenuated the pressor response by 88%. Finally, photostimulation of glutamatergic SFO neurons led to a peak increase in sympathetic nerve activity (n = 2) that was abolished by phenylephrine administration, suggesting that the neuronal circuitry underpinning this response is barosensitive.

Conclusions:
This study demonstrates that the pressor response elicited by glutamatergic SFO neurons is dependent on the PVN and is largely mediated by the sympathetic nervous system via barosensitive neuronal circuitry. Chronic activation of this pathway will be undertaken to determine if an overactivation of this neuronal pathway induces hypertension in normotensive animals.
PS-BPB04-6: PROOF OF BIOLOGICAL ACTIVITY AND EXPLORATION OF EARLY SIGNALING EVENTS OF ANGIOTENSIN-(1–5)
Authors: Silva Igor Maciel Souza , Peluso Antonio Augusto , Jakobsen Lene Andrup , Jensen Pia , Ribeiro Lucas Rodrigues Aguiar , Mortensen Christina , Sumners Colin , Braga Thiago Verano , Larsen Martin Roessel , Steckelings Ulrike Muscha 
Published: January 2023

Introduction:
Recombinant human ACE2 increases the circulating levels of angiotensin-(1–5) [Ang-(1–5)], a peptide thus far regarded as biologically inactive. Since ACE2 is a central component of the protective RAS, we hypothesized that Ang-(1–5) is a new biologically active peptide within this hormonal system.

Objective:
To investigate biological activity and signaling mechanisms of Ang-(1–5).

Methods:
In order to show a biological effect and to test whether Ang-(1–5) signals through the AT2-receptor (AT2R), nitric oxide (NO) release was measured by DAF-FM fluorescence in AT2R transfected (AT2R-CHO) or non-transfected (NT-CHO) CHO cells, treated with Ang-(1–5) or C21 (AT2R agonist, positive control) (0.1 nM to 10 μM) for 15 minutes. Vehicle (cell media) treated cells served as negative control. To investigate Ang-(1–5) signaling patterns, human aortic endothelial cells (HAEC) were treated with vehicle or Ang-(1–5) (1 μM) for 1, 3, 5 or 20 minutes. Proteins were harvested, digested and labeled with TMTpro-16plex. Phosphopeptide enrichment was carried out by TiO2. Samples were subjected to LC-MS/MS analysis and the resulting mass spectra were searched against the human SwissProt database.

Results:
Ang-(1–5) induced a concentration-dependent increase in NO production in AT2-CHO cells (Emax: 65.60 ± 14.02%), thus proving its biological activity. Ang-(1–5) had 69% higher efficacy than the established AT2R agonist C21 (Emax: 38.76 ± 10.24%). Ang-(1–5) seems to signal through the AT2R, because effects on NO release were absent in NT-CHO cells. Treatment of HAEC with Ang-(1–5) significantly modified the phosphorylation status of 831 proteins at 1799 residues. The majority of residues (1079) were dephosphorylated while 729 residues were phosphorylated. Changes in protein phosphorylation in response to Ang-(1–5) occurred at all investigated time points, most of them after 20 minutes. Functional bioinformatic analysis revealed a cluster of proteins involved in cell cycle and cell division regulation.

Conclusion:
This study provides evidence that Ang-(1–5) is an endogenous AT2R agonist with high efficacy towards the AT2R. The early signaling phosphorylation pattern resembles those of other protective RAS agonists, such as C21 and Ang-(1–7).
PS-BPB04-9: DEVELOPMENT AND VALIDATION OF NEW METHOD TO MEASURE PLASMA ALDOSTERONE AND RENIN ACTIVITY USING LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETER (LC-MS/MS)
Authors: Hee Daryl Kim Hor , Tan Yi Ting , Aw Tar Choon , Puar Troy 
Published: January 2023

Objective:
Measurement of aldosterone, renin, and aldosterone-renin ratio (ARR), are used to diagnose primary aldosteronism (PA). Immunoassays used to measure plasma aldosterone yield higher results compared to tandem mass spectrometry (LC-MS/MS). This can lead to false positive diagnosis of PA, and potential for unnecessary adrenal vein sampling. We report our development and validation of a novel test using LC-MS/MS, with reduced run time, for measurement of plasma aldosterone concentration (PAC) and plasma renin activity (PRA).

Design and method:
The detection and quantification of plasma aldosterone and angiotensin 1 (product for the calculation of renin activity) in the mass spectrometer was based on the multiple reaction monitoring of precursor to fragment ions under negative and positive ionization modes, respectively. Validation of the tests were based on the CLSI (Clinical & Laboratory Standards Institute) guidelines. Concurrent measurement for 190 samples were done on our instrument, and on a commercial lab using LC-MS/MS (Mayo Laboratories, USA) for comparison.

Results:
Our measurements of PAC and PRA was sensitive with a limit of quantification at 3 ng/dL and 0.25 ng/ml/h, respectively, and analytical run times were 6.5 min and 3.5 min, respectively. Both aldosterone and angiotensin 1 calibration curves had excellent linearity in the ranges of 3 - 300 ng/dL and 0.25 - 25 ng/ml/h, respectively, with coefficient of correlation, r2 > 0.994. The intra and inter day accuracy of aldosterone and angiotensin 1 were in the range of 95–110% with CV of < 13%. Reference ranges were established for our local population. Compared with the other LC-MS/MS laboratory, there was good correlation for PAC, r2 = 0.90, 95%CI: 0.86 - 0.92, and PRA, r2 = 0.81, 95% CI: 0.75 - 0.86, using 190 samples. In addition, amongst those with low aldosterone < 6ng/dL, there was good agreement between both labs, with 95 of 122 (77.9%) samples having low aldosterone using both methods, which has been shown to be an ideal threshold to exclude the diagnosis of PA.

Conclusions:
Our laboratory has developed and validated a sensitive and accurate LC-MS/MS test for PAC and PRA with shortened run time, for the diagnosis PA.
PS-BPB05-2: ENDOTHELIN A RECEPTOR BLOCKADE INCREASES RENOPROTECTION IN EXPERIMENTAL MODEL OF CONGESTIVE HEART FAILURE COMBINED WITH CHRONIC KIDNEY DISEASE
Authors: Vaneckova Ivana , Kala Petr , Vanourkova Zdenka , Skaroupkova Petra , Cervenka Ludek 
Published: January 2023

Objective:
Congestive heart failure (CHF) represents an extreme burden to the public healthcare worldwide. The incidence and prevalence of chronic kidney disease (CKD) is also increasing and CKD is known as one of the strongest risk factors for the development of CHF. CHF coexists with CKD in approximately half of CHF patients and results in a poor survival rate. The aim of this study was therefore to examine if the addition of endothelin type A (ETA) receptor antagonist atrasentan (5 mg/kg/day) to the angiotensin-converting enzyme inhibitor (ACEi trandolapril; 2 mg/L) will bring additional beneficial effects.

Design and Methods:
CKD was induced by 5/6 renal mass reduction (5/6 NX) at the age of 8 weeks and CHF was elicited by volume overload achieved by the creation of aortocaval fistula (ACF) four weeks later in normotensive Hannover Sprague Dawley rats. The follow-up was 24 weeks after the first intervention (5/6 NX). The treatment regimens were initiated 6 weeks after 5/6 NX and 2 weeks after ACF.

Results:
The final survival in untreated group was 15%. The treatment with ETA receptor antagonist alone or ACEi alone and the combined treatment improved the survival rate to 64%, 71% and 75%, respectively, with no significant difference between the combination and either single treatment regimen. At the end of the study, atrasentan or trandolapril given alone partially reduced albuminuria, but had no effect on albumin/creatinine ratio. In contrast, both parameters were substantially improved by the combined treatment which caused ∼45 % reduction in albuminuria. Moreover, it also reduced markers of renal damage.

Conclusions:
Our results demonstrated that the treatment with ETA receptor antagonist attenuated the mortality related to CKD or CHF and that the addition of ETA receptor antagonist to the standard blockade of renin angiotensin system by ACEi exhibited additional renoprotective actions.
PS-BPB06-1: GLOMERULAR HEMODYNAMIC CHANGES WITH SGLT2 INHIBITION IN TYPE 2 DIABETIC RATS USING IN VIVO IMAGING
Authors: Tatsugawa Rie , Wada Yoshihisa , Kidokoro Kengo , Kashihara Naoki 
Published: January 2023

Background and aim:
Clinical trials have shown that sodium-glucose co-transporter 2 inhibitor (SGLT2i) not only reduce the risk of heart failure and cardiovascular events in people with type 2 diabetes, but also inhibit DKD progression. Improvement of glomerular hyperfiltration via tubuloglomerular feedback (TGF) is considered to be one of the possible pathways for renal protection with SGLT2i in DKD. However, the mechanisms responsible for kidney protection, especially in the setting of T2D, remain incompletely understood. In previous work, we established a method for measuring single nephron GFR (SNGFR) in mice using in vivo imaging and found that the adenosine/adenosine A1 receptor (A1aR) pathway is central for tubuloglomerular feedback (TGF) signaling and the pathogenesis of glomerular hyperfiltration in animal models of type 1 diabetes (Kidokoro K. et al. Circulation 2019). The mechanism of development of glomerular hyperfiltration, and improvement of glomerular hyperfiltration by SGLT2i may, however, be different in type 1 vs type 2 diabetes and related DKD. However, the detailed regulatory mechanism of GFR have not yet been elucidated in DKD related to T2D.

Methods:
Zucker lean (ZL) rats and Zucker diabetic fatty (ZDF) rats were used. In the first experiment, SNGFR and diameters of glomerular afferent/efferent arterioles were measured in both groups. Next, we examined the change of SNGFR and diameters of glomerular afferent/efferent arterioles, as well as urinary excretions of glucose and sodium in ZDF after a single-dose administration of SGLT2i (luseogliflozin; 10 mg/kg, gavage) for 120 minutes, which generated the following three groups: SGLT2i group, SGLT2i + adenosine A1 receptor (A1aR) antagonist (8-cyclopentyl-1,3-dipropylxanthine, 1 mg/kg) group, and insulin group.

Results:
SNGFR, afferent and efferent arteriole diameter were significantly increased in ZDF compared to ZL, with greater afferent vs. efferent dilatation. SGLT2i treatment resulted in correction of afferent arteriole vasodilation, with post-SGLT2i diameter that was comparable to baseline levels (p > 0.05). Post-SGLT2i SNGFR was also similar compared to baseline levels. Changes in efferent arteriole diameter were not significant. The inhibitory effect on hyperfiltration by SGLT2i was abolished by the concomitant use of an A1aR antagonist. In contrast, urinary sodium and urinary glucose in SGLT2 inhibitor group increased significantly regardless of adenosine A1 receptor antagonist use. In response to SGLT2i, urinary adenosine and PGE2 increased significantly (p > 0.05), while urine NOx did not change.

Conclusion:
Regulation of afferent arteriolar tone with SGLT2i is dependent on adenosine/ A1aR signaling an experimental model of T2D related hyperfiltration, regardless of ARB use.
PS-BPB10-4: CREATION AND CHARACTERIZATION OF THE UBIQUITIN LIGASE KLHL3-S433A KNOCK-IN MICE
Authors: Ishizawa Kenichi , Kaseda Ken , Yamazaki Osamu , Shibata Shigeru 
Published: January 2023

Background and aim:
Kelch-like 3 (KLHL3) is a component of an E3 ubiquitin ligase complex that regulates blood pressure and electrolyte homeostasis. Mutation and inactivation of KLHL3 are known to cause hypertension and hyperkalemia mainly due to the impaired with-no-lysine (WNK) degradation and increased Na-Cl cotransporter (NCC) activity in the kidney. Previously, we have reported that angiotensin II (Shibata et al. PNAS 2014), potassium deficiency, and impaired glucose tolerance (Ishizawa et al. BBRC 2016 and JASN2019) can inactivate KLHL3 by altering phosphorylation at S433 in the Kelch-domain. We and others have also shown that decreased KLHL3 expression is also involved in the pathology of hypertension and hyperkalemia. The current study was designed to demonstrate the physiological importance of S433 in KLHL3 function in vivo in mice.

Method:
Using CRISPR/Cas9, S433 of KLHL3 was substituted to alanine in C57BL/6 mice (KLHL3-S433A Knock in mice: KI). Creation of KI was confirmed by DNA sequencing and RFLP analysis. Blood pressure was evaluated by telemetry, and blood gas was analyzed by iSTAT. Levels of KLHL3, WNK1/4, and NCC in the kidney were determined by Western blot analysis. Gene expression of KLHL3, WNK1 and WNK4 was evaluated by real time RT-PCR.

Results:
On a normal salt diet, blood pressure levels were similar between WT and KI; however, KI mice showed significant increase in response to a high salt diet compared with WT on the same diet. KI mice showed a significant increase in plasma potassium levels and a significant decrease in bicarbonate levels already at baseline, confirming that S433 in KLHL3 is also critical in regulating potassium and acid/base homeostasis. Urinalysis indicated that the increase in plasma potassium levels is mainly attributable to the decreased potassium excretion. Of interest, we found that total KLHL3 abundance was drastically reduced by Western blot analysis, which was accompanied by the increase in KS-WNK1 and WNK4 and an increase in NCC both. Real time RT-PCR showed that gene expression of Klhl3, as well as those of Wnk1 and Wnk4, was unchanged, indicating that S433A substitution promotes degradation of KLHL3, leading to hyperkalemia and increased salt sensitivity.

Conclusion:
The study confirmed the critical importance of S433 in KLHL3 in controlling the ubiquitin ligase function and electrolyte homeostasis in vivo. It also indicates that the modification of KLHL3-S433 influences the protein abundance without affecting gene transcription.
PS-BPC02-8: EARLY VASCULAR AGING IN HYPERTENSION IS A PREDICTOR OF INCIDENT DIABETES
Authors: Terentes-Printzios Dimitrios , Christopoulou Georgia , Korogiannis Lampros , Ioakeimidis Nikolaos , Aznaouridis Konstantinos , Dima Ioanna , Gardikioti Vasiliki , Oikonomou Dimitrios , Gkini Konstantia P. , Solomou Eirini , Tsioufis Konstantinos , Vlachopoulos Charalambos 
Published: January 2023

Objective:
Diabetes is associated with vascular aging and arteriosclerosis. Arterial stiffness can be used to assess vascular aging and is an independent predictor of cardiovascular events and mortality. However, the role of arterial stiffness in the incidence of diabetes and especially in patients under treatment with statins has not been investigated. We sought to investigate the role of carotid-femoral pulse wave velocity (PWV) in the prediction of incident diabetes in hypertensives and hypertensives under treatment with statins.

Design and method:
376 never treated hypertensives (mean age 53 ± 13 years, 175 females) without a known history of diabetes or cardiovascular disease at baseline, were included in the study. Markers of target organ damage [carotid femoral PWV, aortic augmentation index (AIx), and estimated glomerular filtration rate (eGFR)] were evaluated at baseline in all patients. eGFR was estimated using the Cockcroft Gault formula. Hemoglobin A1c (HbA1c) and blood glucose were measured in venous blood samples. Patients were followed up prospectively for the incidence of diabetes as defined by the American Diabetes Association criteria. One hundred and thirty-nine patients received statins during follow up.

Results:
During a median of 13.5 years of follow up, 46 patients (12.2%) patients were diagnosed with diabetes. In multivariable logistic regression analysis, only higher PWV (Odds Ratio [OR] = 1.271, 95% Confidence intervals [CI]: 1.002 to 1.612, P = 0.048) was associated with a higher risk of incident diabetes, which was independent of age, sex, pulse pressure, HbA1c, fasting blood glucose and eGFR (Figure). In further analysis, receiver operating characteristic (ROC) curves were generated to evaluate the ability of PWV to discriminate subjects with diabetes. The area under the curve (AUC) and 95% CIs of the ROC curve was AUC = 0.66 (95% CI: 0.58 to 0.74, P = 0.001) for PWV. In the subgroup of patients under treatment with statins, higher PWV (OR = 1.51, 95% CI: 1.09 to 2.11, P = 0.014) was associated with higher risk of incident diabetes, which was independent of the abovementioned confounders. AIx was not associated with the incidence of diabetes.

Conclusions:
Higher arterial stiffness, a marker of early vascular aging, is associated with the incidence of diabetes and also can identify subjects under treatment with statins that are more likely to present with diabetes in the future.
PS-BPC03-2: NIGHTTIME HOME BLOOD PRESSURE WAS ASSOCIATED WITH ADVERSE OUTCOMES IN SUBCLINICAL HEART FAILURE PATIENTS
Authors: Komori Takahiro , Hoshide Satoshi , Kario Kazuomi 
Published: January 2023

Objective:
Nighttime blood pressure (BP) has stronger association with target organ damage and cardiovascular outcome than office BP, 24-hr BP, and awake BP in hypertensives. However, clinical significance of nocturnal BP, especially home nocturnal BP, was not established in heart failure (HF) patients. We hypothesized that nighttime home BP was associated with adverse outcome and investigated the association between nighttime home BP and the outcome in subclinical HF patients.

Design and method:
We analyzed 394 patients with NT-proBNP > 125pg/ml from a practitioner-based population with at least one cardiovascular risk factor who underwent home BP monitoring in the Japan Morning Surge-Home Blood Pressure study. Morning, evening and nighttime home BP were measured by validated, automatic, and oscillometric home BP devices, and cardiovascular events were followed up.

Results:
During a mean follow-up of 4.2 years, 43 cardiovascular events occurred. Morning home SBP, evening home SBP and nighttime home SBP were higher in event group than in non-event group (150 ± 23 vs 140 ± 17 mmHg, p = 0.01, 142 ± 22 vs 131 ± 16 mmHg, p < 0.01, 133 ± 22 vs 125 ± 17 mmHg, p < 0.01, respectively). In multivariate Cox regression analysis adjusting for significant variables, average of evening home SBP, and nighttime home SBP were significant predictors of the outcome (HR 1.02, 95%CI: 1.01–1.04, p = 0.01, HR 1.02, 95%CI: 1.01–1.04, p = 0.03, respectively). When nighttime home SBP of 120mmHg was used as cutoff, nighttime home SBP > 120mmHg was significantly associated with the outcome (HR 2.46, 95%CI 1.06–5.68, p = 0.04).

Conclusions:
Elevated nighttime home BP was significant predictor of cardiovascular outcomes in subclinical HF patients.
PS-BPC03-4: DIAGNOSTIC AGREEMENT OF MASKED UNCONTROLLED HYPERTENSION DETECTED BY AMBULATORY BP AND HOME BP MEASURED BY AN ALL-IN-ONE BP MONITORING DEVICE
Authors: Tomitani Naoko , Hoshide Satoshi , Kario Kazuomi 
Published: January 2023

Objectives:
Masked hypertension is a phenomenon in which office blood pressure (BP) is in the controlled BP range while out-of-office BP measured by ambulatory BP monitoring (ABPM) and home BP monitoring (HBPM) is in the uncontrolled BP range. Out-of-office BP control status may differ if defined by different BP indexes or measured under different conditions (resting or ambulatory), over different time-windows, or with different devices. This study aims to investigate the diagnostic agreement of masked uncontrolled hypertension (MUHT) detected by ABPM indexes (ABPM-MUHT) and HBPM indexes (HBPM-MUHT) using the same all-in-one device.

Methods:
Medicated hypertension patients enrolled in the Home-Activity ICT-based Japan Ambulatory Blood Pressure Monitoring Prospective (HI-JAMP) Study between 2017 and 2020 consecutively underwent office BP monitoring, 24-h ABPM (with 30-min intervals), and 5-day HBPM (twice each morning and evening) using the same device (TM-2441; A&D Company, Tokyo). Controlled BP ranges for each BP index were as follows: office systolic BP [SBP] < 140 mmHg; morning home, evening home, or morning-evening (ME) average SBP < 135 mmHg; 24-h average SBP < 130 mmHg; daytime SBP < 135 mmHg; and nighttime SBP < 120 mmHg.

Results:
Of 2322 patients (males 53.2%, average age 69.2 ± 11.5 years, average office SBP 132.8 ± 18.8 mmHg), the prevalences of ABPM-MUHT and HBPM-MUHT were 11.0% and 13.9%, respectively, when MUHT was defined only by 24-h average SBP or ME average SBP. Among the 445 patients with any MUHT according to this definition, the diagnostic agreement of MUHT detected by ABPM and HBPM was 29.7%. The prevalences of HBPM-MUHT defined by morning SBP and evening SBP were 22.4% and 11.6%, respectively, and those of ABPM-MUHT defined by daytime SBP and nighttime SBP were 11.8% and 13.8%. When MUHT was defined by any time-window index (ABPM: daytime, nighttime, and 24-h average SBP; HBPM: morning, evening, ME average SBP), the prevalences of ABPM-MUHT and HBPM-MUHT were 19.8% and 25.0%, respectively, and the diagnostic agreement of MUHT detected by ABPM and HBPM in patients with any MUHT (n = 743) increased to 40.1%.

Conclusion:
Even in treated patients with well-controlled office BP, the prevalence of MUHT in the morning and nighttime was high. Assessment of BP control status only by ME average or 24-h average SBP, without considering specific time-windows, might underestimate cardiovascular risk.