bluuebunny/crossref_metadata_2025_split huggingface dataset:
PS-BPB05-1: DUAL DISRUPTION OF ENDOTHELIAL NITRIC OXIDE SYNTHASE AND APOE GENE ACCELERATES KIDNEY FIBROSIS AND AGING AFTER INJURY
Authors: Kishi Seiji , Nishimura Kenji , Kashihara Naoki 
Published: January 2023

Objective:
Advances in medical science have made it possible to control diseases such as cancer, autoimmune diseases, and infectious diseases, extending the lifespan of affected people around the world. On the other hand, the number of patients with Chronic Kidney Disease (CKD) is increasing worldwide, and hypertension is a key factor. Suppression of CKD is important for extending healthy life expectancy. In this study, we will clarify the effects of the interaction between hypertension and atherosclerosis on renal fibrosis and aging.

Design and method:
Wild type (WT) mouse, apolipoprotein E-/- (ApoE KO) mouse, and endothelial nitric oxide synthase (eNOS)-/- ; ApoE-/- (DKO) mouse were obtained by crossing eNOS+/- mouse and ApoE+/- mouse. Unilateral ureteral obstruction (UUO) was performed on 8–10 weeks old male mice after blood pressure and lipid profile were measured. Mice were sacrificed 10 days after UUO. The degree of renal tubular injury, fibrosis and kidney senescence were evaluated.

Results:
DKO mice exhibited the most pronounced blood pressure and lipid abnormalities among the three groups, with a significant elevation of blood pressure, total cholesterol, and low HDL cholesterol compared to WT mice. DKO mice manifested elevated blood pressure, higher total cholesterol and lower HDL than WT mice. Compared with WT mice, ApoE KO and DKO mice showed sustained kidney injury molecule-1 expression and kidney fibrosis was significantly higher in ApoEKO and DKO mice. mRNA expression of genes related to kidney fibrosis was the highest in DKO mice. The mRNA expression of Zinc alpha2 Glycoprotein and heme oxygenase-1 were significantly decreased in DKO mice. Furthermore, mRNA expression of p53, p21 and p16 were increased both in ApoEKO and DKO mice, with DKO mice being the highest. Senescence associated beta-gal positive tubule area was significantly increased in DKO. Increased DNA damage and target of rapamycin autophagy spatial coupling compartments (TASCCs, Sci Transl Med. 2019;11(476):eaav4754. J Clin Invest. 2019;129(11):4797- 4816.) formation was found in DKO mice.

Conclusion:
Mice at high risk for cardiovascular disease developed kidney fibrosis and aging even in the young mice after injury. Vulnerability to oxidative stress due to endothelial cell dysfunction promotes fibrosis and aging. Managing lifestyle-related diseases including hypertension from a young age is important for CKD prevention.
PS-BPB05-3: SERINE PROTEASE INHIBITION EXERTS AQUARETIC EFFECT AND DECREASES URINARY EXOSOMAL AQP2 LEVELS
Authors: Kakizoe Yutaka , Iwata Yasunobu , Nakagawa Terumasa , Deng Qinyuan , Adachi Masataka , Miyasato Yoshikazu , Nakagawa Miyuki , Nagayoshi Yu , Nishiguchi Kayo , Narita Yuki , Izumi Yuichiro , Kuwabara Takashige , Mukoyama Masashi 
Published: January 2023

Objective:
Serine proteases (SPs) play pivotal roles in various physiological processes. In the kidney, it is reported that the epithelial sodium channel (ENaC) is activated via the proteolytic cleavage of α and γ subunits by SPs. We previously reported that a synthetic SP inhibitor camostat mesilate (CM) suppressed the cleavage of γENaC, thereby decreasing sodium reabsorption and blood pressure in aldosterone-infused rats. These results suggest the important roles of SPs within the kidney in the regulation of sodium homeostasis and blood pressure. Here, we aimed to identify novel physiological roles of SPs in the renal tubule and understand the mechanism of actions of SP inhibitors by administering CM into healthy rats.

Design and method:
Six-week-old male Sprague-Dawley rats were divided into control and CM groups. CM group rats were subcutaneously implanted with sustained-release pellets of CM (14 mg/day). After 24 h urine collection was performed, rats were sacrificed at day 7 to obtain blood and kidney samples. Urine volume was measured during the experimental period, and blood and urinary electrolytes, blood vasopressin, and renal tissue osmolality were measured. Protein expression of SPs and aquaporin-2 (AQP2) was analyzed by Western blotting.

Results:
CM significantly increased urine volume by approximately two-fold throughout the experimental period [urine volume (mL/day): control, 15.5 ± 2.4; CM, 30.6 ± 5.8], in a urinary sodium and osmolyte excretion-independent manner, which indicated that free water excretion had occurred. The vasopressin, potassium, and calcium levels in the blood and osmolality in the inner medulla, which affect free water reabsorption in the renal tubule, remained unchanged after CM administration. CM decreased urinary exosomal AQP2 excretion, suggesting that the AQP2 activity was suppressed in the collecting duct. These changes were reversed by the infusion of desmopressin. Such water diuretic action caused by CM was independent of its action on prostasin and TMPRSS4, which are serine proteases associated with sodium and water homeostasis, respectively.

Conclusion:
Our results revealed the association of SP inhibition with free water diuresis in the kidney and demonstrated that CM administration resulted in the downregulation of AQP2. The current findings suggest that SP inhibitors could become a new class of aquaretic drugs.
PS-BPB05-7: UPREGULATION OF ALDOSTERONE SYNTHASE INDUCED BY LUTEINIZING HORMONE IN HUMAN ADRENOCORTICAL CELLS
Authors: Morishima Noriko , Morimoto Satoshi , Watanabe Daisuke , Kato Yoichiro , Shibata Noriyuki , Ichihara Atsuhiro 
Published: January 2023

Objective:
Plasma aldosterone concentration (PAC) is increased in stages of elevation of luteinizing hormone (LH) during the luteal phase of the ovarian cycle, menopause, or pregnancy. PAC is elevated in patients with polycystic ovary syndrome who have elevated circulating concentration of luteinizing hormone (LH). In addition, stimulation of aldosterone production by LH has been reported. In the present study, we investigated the mechanism by which LH increases aldosterone production in human adrenocortical cells.

Design and Methods:
Immunohistochemical staining for CYP11B2, the aldosterone synthase, and LH receptor were performed using autopsy adrenal gland specimens. Changes in mRNA or protein expressions of CYP11B2 after treatment with LH, angiotensin (Ang) II, or LH+Ang were compared in human HAC15 adrenocortical cells. Effects of an Ang receptor blocker (ARB), adenylyl cyclase (AC) inhibitor, cyclic AMP response element binding protein (CREB) inhibitor, protein kinase A (PKA) inhibitor, or β;-catenin inhibitor on the increase in CYP11B2 mRNA expression induced by LH+Ang ll were investigated in these cells. In addition, effects of an AC agonist on CYP11B2 mRNA expression were investigated in these cells.

Results:
Immunostaining for LH receptor was detected in cells positive for CYP11B2 in adrenal granulosa and fasciculata region in the human autopsy specimens. CYP11B2 mRNA and protein expressions were significantly increased after treatment with LH+Ang ll than LH or Ang ll alone in HAC cells. Elevation of CYP11B2 mRNA expression induced by LH+Ang ll was completely suppressed by the ARB, AC inhibitor, or CREB inhibitor and was significantly reduced by the PKA inhibitor or β;-catenin inhibitor in these cells. CYP11B2 mRNA expression was significantly increased by the AC agonist.

Conclusion:
The present study showed that LH receptor is expressed in human adrenal cortical region and that LH augments CYP11B2 expression via AC-CREB pathway in the presence of Ang ll in human adrenocortical cells.
PS-BPB06-3: EXAMINATION OF THE EFFECTIVENESS OF DIABETIC NEPHROPATHY TREATMENT TARGETING CHREBP ACTIVITY
Authors: Suzuki Susumu , Yokoyama Atsushi , Okamoto Koji , Takahashi Shinichiro , Iwabuchi Yoshiharu , Sugawara Akira 
Published: January 2023

Introduction:
Diabetic nephropathy (DN) is the main cause of chronic kidney disease (CKD), and is characterized by elevated urinary albumin to creatinine ratio (UACR) and serum creatinine, and decreased glomerular filtration rate. Anti hypertensive agents are used for DN treatment, however, the effects of these drugs are insufficient, and the establishment of more effective treatment methods is desired.
Carbohydrate responsive element binding protein (ChREBP) is one of the factors reported to be involved in the onset and progression of DN. Furthermore, whole body ablation of ChREBP suggested that the progression of DN was suppressed in the rodent model. On the other hand, the effectiveness of pharmaceutical ChREBP suppression is unclear, thus, we performed administration experiment to DN model mice using ChREBP activity inhibitor and examined the effects of pharmaceutical ChREBP activity suppression on renal function of diabetic mice.

Results:
First, we established high-throughput screening system capable of detecting ChREBP activity and performed screening of Tohoku university compound library. The result compound, D-532, was ChREBP activity specific inhibitor without cell toxicity. D-532 directly binds to DNA-binding domain of ChREBP, and inhibits the transcriptional activity. We next performed administration experiment using type 1 DN model mice. D-532 treatment group showed an improvement tendency of the low body weight compared with that of vehicle treatment group, while it had no effect on blood glucose level. About renal functions, both of UACR and serum creatinine were significantly lower in D-532 treatment group. Importantly, renal tissues such as glomeruli and renal tubules were preserved in D-532 treatment group compared with in vehicle treatment group. In vivo as well, D-532 suppressed ChREBP target genes mRNA expression, and RNA-sequence analysis with mice renal cortex RNA at the end of D-532 administration experiment indicated that D-532 suppressed excessive oxidative stress through the ChREBP activation under diabetes.

Conclusion:
These results indicate that ChREBP suppression is effective on DN rodent model, and ChREBP activity inhibitor D-532 may have potential as a new treatment for DN in the future.
PS-BPB06-4: EXERCISE TRAINING AMELIORATES RENAL OXIDATIVE STRESS IN RATS WITH CHRONIC KIDNEY DISEASE
Authors: Yamakoshi Seiko , Nakamura Takahiro , Mori Nobuyoshi , Kohzuki Masahiro , Mori Takefumi , Ito Osamu 
Published: January 2023

Background:
The renoprotective effects of long-term exercise in chronic renal failure have been reported. However, the details of the mechanism remains unclear. This study investigated the renal protective effect of long-term exercise on renal oxidative stress in a rat model of chronic renal failure.

Methods:
Male Sprague-Dawley rats were divided into Sham, 5/6 nephrectomy + sedentary, and 5/6 nephrectomy + exercise training groups. 5/6 nephrectomy + exercise training group underwent treadmill running for 12weeks. Systolic blood pressure and urinary protein were measured every 2 weeks. Renal function, renal histology and oxidative stress-related factors were examined after completion of the exercise protocol.

Results:
5/6 nephrectomized rats exhibited hypertension, proteinuria, renal dysfunction glomerulosclerosis, renal interstitial fibrosis, and exercise training ameliorated them. 5/6 nephrectomy increased urinary malondialdehyde, renal cortical NADPH oxidase and xanthine oxidase activities and exercise training inhibited the 5/6 nephrectomy-increased urinary malondialdehyde, renal cortical NADPH oxidase and xanthine oxidase activities. Plasma malondialdehyde was not affected by 5/6 nephrectomy or exercise.

Conclusions:
In rats with chronic kidney disease, long-term exercise prevented hypertension, renal dysfunction, glomerulosclerosis, and renal interstitial fibrosis. Mechanism for the antihypertensive and renal protective effects of long-term exercise in chronic kidney disease may involve the reduction of renal oxidative stress.
PS-BPB06-6: RENAL INJURY ASSOCIATED WITH ARGININE VASOPRESSIN INDUCTION IN A RAT MODEL OF PREECLAMPSIA
Authors: Ramdin Sapna , Baijnath Sooraj , Naicker Thajasvarie , Govender Nalini 
Published: January 2023

Objective:
Worldwide, preeclampsia (PE) accounts for 2–8% of pregnancies and is a main cause of maternal and fetal morbidity and mortality. In vivo murine studies confirm the use of arginine vasopressin (AVP) in replicating the clinical symptoms of PE development, suggestive of a role for PE prediction. The renal effects that AVP may have in PE remains elusive, hence this study aimed to explore the physiological effects of AVP on the kidneys of pregnant rats in an effort to determine its role in PE development.

Design and method:
Twenty-four female Sprague Dawley rats (non-pregnant saline, pregnant saline, non-pregnant AVP and pregnant AVP) were surgically implanted with ALZET mini osmotic pumps infusing with either saline or AVP for 18 days. Blood pressure measurements and urine samples were taken on gestational days (GD) 8, 14 and 18. Placental and fetal weights were recorded following sacrifice on GD18. Urinary levels of kidney injury molecule-1 (KIM-1) were determined using the rat kidney toxicity immunoassay panel 1 (#RKTX1MAG-37K). Immunohistochemistry was used to determine the expression of KIM-1 in rat kidney tissue.

Results:
Systolic [143.00 (139.67–144.00) mmHg vs 125.00 (124.00–127.00) mmHg; p < 0.05] and diastolic blood pressures [103.50 (100.33–106.88) mmHg vs 81.00 (80.00–87.00) mmHg; p < 0.001] were significantly higher in the pregnant AVP vs pregnant saline group. Urinary protein levels in pregnant AVP rats [0.45 (0.42–0.48) g/L] were significantly elevated in comparison to pregnant saline rats [0.24 (0.21–0.32) g/L; p < 0.05]. Pregnant AVP rats also demonstrated a significant decrease in individual placental [0.39 (0.31–0.46) g vs 0.59 (0.55–0.63) g; p < 0.05] and pup weights [1.57 (1.45–1.67) g vs 1.79 (1.70–1.85) g; p < 0.05] in comparison to pregnant saline rats. Urinary KIM-1 levels are significantly elevated in the pregnant AVP (0.53 ± 0.05 ng/mL) vs pregnant saline group (0.17 ± 0.02 ng/mL; p < 0.001). Qualitative analysis revealed that KIM-1 expression is markedly upregulated on the apical membrane surface of epithelial cells of the proximal convoluted tubules in AVP treated rats.

Conclusions:
AVP infusion increases blood pressure, urine protein levels and urinary and tissue expression of KIM-1. Our findings indicate that there is significant kidney injury associated with AVP treatment, indicative of the utility of this model in studying the mechanisms driving renal damage in PE.
PS-BPB06-9: PREVALENCE OF KCNJ5 MUTANT ALDOSTERONE-PRODUCING ADENOMAS AMONG MALAYSIAN PATIENTS WITH PRIMARY ALDOSTERONISM
Authors: Rizam Nurfatin Syazreen Mohd , Mustangin Muaatamarulain , Tan Geok Chin , Sukor Norlela , Saparuddin Fatin , Pauzi Fatin Athirah , Nazri Siti Khadijah Syed Muhammad , Nasruddin Azraai Bahari , Azizan Elena Aisha 
Published: January 2023

Objective:
To verify the prevalence of KCNJ5 mutant aldosterone-producing adenomas (APAs) among primary aldosteronism (PA) patients from a tertiary center in Malaysia

Design and method:
A total of 80 adrenal samples from adrenalectomies performed between 2010 up to 2020 at Hospital Putrajaya, Malaysia were included in this study. Of the adrenals blocks received, DNA samples from 74 APAs identified by CYP11B2 immunohistochemistry (IHC) staining were sent for KCNJ5 mutation analysis. Gender, ethnicity and age at adrenalectomy were compared between genotypes. Data were statistical analyzed using the Fisher Exact test.

Results:
28 of the 74 (37.8%) APAs had a KCNJ5 mutation. The most common KCNJ5 variant was G151R (50%), followed by L168R (39.3%) and T158A (10.7%). Of all the mutant APAs, 19/28 (67.9%) were from female PA patients whereas among controls 26/46 (56.5%) were females. No significant association between KCNJ5 mutation status and gender were found. The majority of mutant APAs were from Malay PA patients (n = 17), followed by Chinese PA patients (n = 11) which follows the whole cohort demography. Thus, ethnicity was not found to be significantly associated with KCNJ5 mutation status. In this cohort, no significant differences in age at of adrenalectomy by KCNJ5 mutation status was seen. Adrenalectomy procedure was performed most frequent between the age of 37–46 years old (n = 30), followed by 47–56 (n = 16), 26–35 (n = 12), 57–66 (n = 11) and 67–76 (n = 2); in 3 patients this information was not known.

Conclusions:
A similar prevalence of KCNJ5 mutant APAs as found in a previous study performed at the National University of Malaysia Medical Centre between 2000 and 2015 verifies the lower prevalence of KCNJ5 mutant APAs in Malaysian PA patients (37.8%) compared to that observed in other Oriental cohorts (up to 70%). Although, the majority of mutant APAs were from female PA patients (67.9%), no gender bias according to KCNJ5 mutation status was found as the majority of PA patients in this cohort were females (60.8%).
PS-BPB06-10: DISTRIBUTION OF CACNA1D SOMATIC MUTATIONS AMONG CYP11B2 IMMUNOHISTOCHEMISTRY-GUIDED SAMPLES OF ALDOSTERONE-PRODUCING ADENOMA (APA)
Authors: Pauzi Fatin Athirah , Azizan Elena Aisha , Mustangin Muaatamarulain , Tan Geok Chin , Sukor Norlela , Ryska Ales , Ceral Jiri , Solar Miroslav , Nasruddin Azraai Bahari 
Published: January 2023

Objectives:
Past studies in Afro-Caribbean primary aldosteronism (PA) patients have found CACNA1D mutations to be the most common aldosterone-driving mutation among their cohort; a striking difference to Caucasian and Oriental cohorts where KCNJ5 mutations are reported to be the most common. However not many studies had used aldosterone synthase (CYP11B2) immunohistochemistry (IHC)-guided sequencing approach of the whole CACNA1D gene, performing instead unguided adenoma sampling and hotspot sequencing. We thus performed CYP11B2 IHC-guided whole gene sequencing on APAs from an Oriental cohort (Malaysia) and a Caucasian cohort (Czech Republic) for comparison.

Design and method:
Surgically removed adrenals from 100 Malaysian and Czech PA patients with unilateral disease were studied. Genomic DNA was isolated from adenoma FFPE sections that had a positive CYP11B2 IHC staining and somatic mutations analysis were performed by next generation sequencing (NGS) of known aldosterone-driver genes. Predicted pathogenic mutations were verified by Sanger Sequencing.

Results:
Out of 100 FFPE samples sent for NGS, 36 were excluded due to the quality of the data. In total 58 of 64 APAs (87.5%) had a pathogenic aldosterone-driver somatic mutation. KCNJ5 mutations were the most common (n = 37, 57.8%), followed by CACNA1D (n = 11, 20.3%), ATP1A1 (n = 6, 9.4%), ATP2B3 (n = 3, 4.7%) and CTNNB1 (n = 2, 3.1%) mutations. Of particular interest, one APA sample had two pathogenic somatic mutations, CACNA1D p.Val1373Met and CTNNB1: p.Thr41Ala. SIFT and Polyphen2 scores predicted both mutations to be deleterious. Both mutations have previously been reported separately to cause APAs. The CACNA1D/CTNNB1 mutant APA, was solitary, with homogeneous CYP11B2 expression while negative for CYP17A1 expression. The APA was adrenalectomized from a 53-year-old male Czech patient. The majority of patients with APAs harboring CACNA1D mutations were male (n = 8, 72.7%) and from the Czech cohort (n = 7, 64%). There were no significant differences in age at adrenalectomy between patients harboring a CACNA1D mutant APA (45 ± 3) and those with a non-CACNA1D mutant APA (48 ± 2).

Conclusions:
Solitary ZG APAs can harbor both a CACNA1D and a CTNNB1 mutation. The prevalence of CACNA1D in this CYP11B2-IHC guided study is consistent with previous results reported in Oriental and Caucasian cohorts. Although CACNA1D is not the top genetic alteration responsible for APAs in these cohorts, CACNA1D may still be a potential key player in idiopathic hyperaldosteronism and aldosterone-producing micronodules. Thus further study is required to characterize the distribution of CACNA1D pathogenic mutations in bilateral PA.
PS-BPB07-2: AFFERENT RENAL NERVES CONTRIBUTE TO LEFT VENTRICULAR DYSFUNCTION THROUGH HYPOTHALAMIC VASOPRESSIN-MEDIATED CENTRAL SYMPATHOEXCITATION IN HYPERTENSIVE HEART FAILURE MODEL
Authors: Ikeda Shota , Shinohara Keisuke , Kashihara Soichiro , Matsumoto Sho , Yoshida Daisuke , Nakashima Ryosuke , Ono Yoshiyasu , Tsutsui Hiroyuki 
Published: January 2023

Objective:
Afferent renal nerves may contribute to the development and progression of heart failure, which, however, has not been well investigated. This study aimed to examine whether the afferent renal nerve input contributes to hypertensive cardiac dysfunction.

Design and method:
Male Dahl salt-sensitive rats were fed 0.3% low-salt diet (LS) or 8% high-salt diet (HS) from 6 weeks of age. HS-fed rats were divided into selective afferent renal denervation (HS-ARDN) or sham operation (HS-Sham) at 9 weeks. The effects of ARDN on central sympathetic regulation were investigated at 12 weeks. In another cohort, its effects on the cardiac phenotypes were evaluated at 16 weeks, with adding total renal denervation (HS-TRDN) group.

Results:
At 12 weeks, systolic blood pressure (SBP) measured by tail-cuff method was significantly increased in HS-Sham compared to LS, while there was no significant difference between HS-Sham and HS-ARDN. Plasma norepinephrine concentration was increased in HS-Sham compared to LS and was attenuated in HS-ARDN. The c-Fos expression, a neuronal activation marker, in presympathetic neurons within hypothalamic paraventricular nucleus (PVN) and rostral ventral lateral medulla (RVLM) was also increased in HS-Sham and attenuated in HS-ARDN, which was in parallel with the c-Fos expression in vasopressin-expressing neurons within PVN. At 16 weeks, SBP was similarly increased in HS-Sham, HS-ARDN, and HS-TRDN compared to LS. Although the parameters of left ventricular (LV) hypertrophy, such as LV wall thickness by echocardiography and LV weight, were significantly increased in HS-Sham, HS-ARDN, and HS-TRDN compared to LS, there was no significant difference between HS-fed three groups. Echocardiographic LV fractional shortening was significantly decreased in HS-Sham compared to LS, and was attenuated to the same extent in HS-ARDN and HS-TRDN. Histological LV fibrosis determined by Masson trichrome staining and mRNA expression of ANP and CTGF in LV were significantly increased in HS-Sham compared to LS; these changes were similarly ameliorated in both HS-ARDN and HS-TRDN.

Conclusion:
The input from renal afferent nerves contributes to sympathoexcitation and cardiac dysfunction independently of SBP in Dahl salt-sensitive hypertensive heart failure model. Vasopressin production in PVN may be involved in the renal afferents-mediated sympathetic regulation.