Kunihisa Yamaguchi

Adiponectin inhibits insulin-like growth factor-1-induced cell migration by the suppression of extracellular signal-regulated kinase 1/2 activation, but not Akt in vascular smooth muscle cells.

Adiponectin, an adipocyte-derived hormone, has been proposed to show antiatherogenic properties through the inhibitory effects against various growth factors. Insulin-like growth factor-1 (IGF-1) is one of the potent mitogens, which has been considered to play important roles in both atherogenesis and plaque stabilization in accordance to the phase of atherosclerosis. The aim of this study is to elucidate the adiponectin effects on IGF-1-induced cell migration and its intracellular signaling pathways in vascular smooth muscle cells (VSMCs). In this study, we assessed cell migration and several kinase activities in cultured rat aortic smooth muscle cells (RASMCs). Adiponectin pretreatment suppressed IGF-1-induced cell migration and extracellular signal-regulated kinase (ERK)1/2 activation, which is one of the major mediators for IGF-1-induced cell migration. In RASMCs, adiponectin and 5-aminoimidazole-4-carboxamide riboside (AICAR), a 5′-AMP-activated protein kinase (AMPK) activator, stimulated AMPK activation. AMPK activation by AICAR inhibited IGF-1-induced ERK1/2 activation and cell migration in RASMCs. On the other hand, phosphorylation of Akt and Bad, proapoptotic molecules of the Bcl-2 family, which were increased by IGF-1 stimulation, was not diminished by the pretreatment with adiponectin. It was shown that adiponectin inhibited IGF-1-induced VSMC migration through suppression of ERK1/2 activation, which might be implicated in AMPK activation. Furthermore, adiponectin selectively inhibited ERK1/2 pathway, not Akt–Bad pathway, stimulated by IGF-1. From these findings, it was implied that adiponectin suppressed IGF-1-induced VSMC migration and its signaling selectivity.

Dietary doses of nitrite restore circulating nitric oxide level and improve renal injury in l-NAME-induced hypertensive rats

We have reported that pharmacological doses of oral nitrite increase circulating nitric oxide (NO) and exert hypotensive effects in Nomega-nitro-L-arginine methyl ester (L-NAME)-induced hypertensive rats. In this study, we examined the effect of a chronic dietary dose of nitrite on the hypertension and renal damage induced by chronic L-NAME administration in rats. The animals were administered tap water containing L-NAME (1 g/l) or L-NAME + nitrite (low dose: 0.1 mg/l, medium dose: 1 mg/l, high dose: 10 mg/l) for 8 wk. We evaluated blood NO levels as hemoglobin-NO adducts (iron-nitrosyl-hemoglobin), using an electron paramagnetic resonance method. Chronic administration of L-NAME for 8 wk induced hypertension and renal injury and reduced the blood iron-nitrosyl-hemoglobin level (control 38.8 +/- 8.9 vs. L-NAME 6.0 +/- 3.1 arbitrary units). Coadministration of a low dose of nitrite with L-NAME did not change the reduced iron-nitrosyl-hemoglobin signal and did not improve the L-NAME-induced renal injury. The blood iron-nitrosyl-hemoglobin signals of the medium dose and high dose of nitrite were significantly higher than that of L-NAME alone. Chronic administration of a medium dose of nitrite attenuated L-NAME-induced renal histological changes and proteinuria. A high dose of nitrite also attenuated L-NAME-induced renal injury. These findings suggest that dietary doses of nitrite that protect the kidney are associated with significant increase in iron-nitrosyl-hemoglobin levels. We conclude that dietary nitrite-derived NO generation may serve as a backup system when the nitric oxide synthase/L-arginine-dependent NO generation system is compromised.

Low Transscrotal Orchidopexy Is a Safe and Effective Approach for Undescended Testes Distal to the External Inguinal Ring

Objectives: To review the results of a low transscrotal orchidopexy in patients with palpable undescended testes located distal to the external inguinal ring. Methods: Between July 1998 and June 2005, transscrotal orchidopexy with a single low scrotal incision was performed in 32 patients for 49 undescended testes. The indication was an undescended testis that lay distal to the external ring and could be pulled down manually into the scrotum under general anesthesia. Results: All testes that were treated with the low transscrotal approach were successfully fixed in the middle or lower portion of the scrotum. The mean operative time was significantly shorter for the low transscrotal orchidopexy (45.2 min) than for the inguinal orchidopexy (66.6 min) for 107 undescended testes at similar locations. The median follow-up duration was 39.1 months; all testes except 1 (97.7%) were located in a good position within the scrotum and had a good consistency; 1 testis ascended postoperatively and required inguinal orchidopexy. No inguinal hernias or hydroceles occurred after the surgery. Conclusions: Low transscrotal orchidopexy appears to be an excellent alternative to the standard inguinal orchidopexy for undescended testes located distal to the external inguinal ring.

Dual involvement of growth arrest-specific gene 6 in the early phase of human IgA nephropathy.

Background Gas6 is a growth factor that causes proliferation of mesangial cells in the development of glomerulonephritis. Gas6 can bind to three kinds of receptors; Axl, Dtk, and Mer. However, their expression and functions are not entirely clear in the different glomerular cell types. Meanwhile, representative cell cycle regulatory protein p27 has been reported to be expressed in podocytes in normal glomeruli with decreased expression in proliferating glomeruli, which inversely correlated with mesangial proliferation in human IgA nephropathy (IgAN). Methods The aim of this study is to clarify Gas6 involvement in the progression of IgAN. Expression of Gas6/Axl/Dtk was examined in 31 biopsy proven IgAN cases. We compared the expression levels with histological severity or clinical data. Moreover, we investigated the expression of Gas6 and its receptors in cultured podocytes. Results In 28 of 31 cases, Gas6 was upregulated mainly in podocytes. In the other 3 cases, Gas6 expression was induced in endothelial and mesangial cells, which was similar to animal nephritis models. Among 28 podocyte type cases, the expression level of Gas6 correlated with the mesangial hypercellularity score of IgAN Oxford classification and urine protein excretion. It also inversely correlated with p27 expression in glomeruli. As for the receptors, Axl was mainly expressed in endothelial and mesangial cells, while Dtk was expressed in podocytes. In vitro, Dtk was expressed in cultured murine podocytes, and the expression of p27 was decreased by Gas6 stimulation. Conclusions Gas6 was uniquely upregulated in either endothelial/mesangial cells or podocytes in IgAN. The expression pattern can be used as a marker to classify IgAN. Gas6 has a possibility to be involved in not only mesangial proliferation via Axl, but also podocyte injury via Dtk in IgAN.

Complications of Flexible Ureteroscopic Treatment for Renal and Ureteral Calculi during the Learning Curve

Background: The flexible ureterorenoscope (URS) and associated devices have developed rapidly. However, despite its therapeutic benefits, URS may be associated with some complications. To the best of our knowledge, there are no studies discussing the complications of flexURS during the learning curve. Methods: A retrospective review of the records of patients who underwent flexURS from January 2005 to June 2013 was performed. To compare the complications after the introduction of flexURS, patients were divided into four groups based on the surgeons training experience, that is, based on the number of cases performed by the surgeon. A total of 219 cases underwent flexURS. Groups 1, 2, 3, and 4 included 35, 50, 50, and 84 cases, respectively. The complications were classified using the Clavien system (I-IV). Results: The mean operation time and stone-free rate were significantly different (p < 0.001, p = 0.013, respectively). The total complication rates were 13.6, 10, 8.3, and 3.2%, respectively (p = 0.068). The more the surgeons experience, the less was the complication rate. Despite our best efforts, the incidence of urosepsis was not reduced (p = 0.902). Conclusions: To reduce severe complications, it is necessary to have performed about 100 cases. Increased surgeon experience tended to decrease the risk of severe complications, but the incidence of urosepsis was not reduced.

Clinical Significance of Neoadjuvant Combined Androgen Blockade for More Than Six Months in Patients with Localized Prostate Cancer Treated with Prostate Brachytherapy.

Introduction: The aim of this study is to clarify the clinical significance of neoadjuvant combined androgen blockade (CAB) for ≥6 months in patients with localized prostate cancer. Patients and Methods: A total of 431 patients with localized prostate cancer who underwent prostate brachytherapy (BT) with or without neoadjuvant CAB for ≥6 months with mean follow-up time of 64.6 months (range 24-108 months) were evaluated retrospectively. Of those 431, 232 patients received BT in combination with neoadjuvant CAB for ≥6 months. Biochemical recurrence-free rates (BRFRs) in 364 patients with at least 3 years of follow-up were evaluated by log-rank test. Results: BRFR in patients with low-, intermediate- and high-risk prostate cancer were 98.1, 94.2 and 89.1%, respectively. In patients with intermediate-risk prostate cancer only, neoadjuvant CAB was significantly associated with BRFR (p = 0.0468). Especially in patients with intermediate-risk prostate cancer with radiation dose received by 90% of the prostate (D90) <180 Gy, neoadjuvant CAB exerted a favorable impact on BRFR (p = 0.0429). On multivariate analyses, neoadjuvant CAB and D90 were independent predictors of BRFR (p = 0.0061 and p < 0.0001, respectively). Conclusions: Neoadjuvant CAB for ≥6 months has a favorable impact on BRFR in patients with intermediate-risk prostate cancer, particularly in patients with relatively low radiation doses of D90.

The Advantage of a Ureteroscopic Navigation System with Magnetic Tracking in Comparison with Simulated Fluoroscopy in a Phantom Study

PURPOSE To assess whether our ureteroscopic real-time navigation system has the possibility to reduce radiation exposure and improve performance of ureteroscopic maneuvers in surgeons of various ages and experience levels. MATERIALS AND METHODS Our novel ureteroscopic navigation system used a magnetic tracking device to detect the position of the ureteroscope and display it on a three-dimensional image. We recruited 31 urologists from five institutions to perform two tasks. Task 1 consisted of finding three internal markings on the phantom calices. Task 2 consisted of identifying all calices by ureteroscopy. In both tasks, participants performed with simulated fluoroscopy first, followed by our navigation system. Accuracy rates (AR) for identification, required time (T) for completing the task, migration length (ML), and time exposed to simulated fluoroscopy were recorded. RESULTS The AR, T, and ML for both tasks were significantly better with the navigation system than without it (Task 1 with simulated fluoroscopy vs with navigation: AR 87.1 % vs 98.9%, P=0.003; T 355 s vs 191 s, P<0.0001; ML 4627 mm vs 2701 mm, P<0.0001. Task 2: AR 88.2% vs 96.7%, P=0.011; T 394 s vs 333 s, P=0.027; ML 5966 mm vs 5299 mm, P=0.0006). In both tasks, the participants used the simulated fluoroscopy about 20% of the total task time. CONCLUSIONS Our navigation system, while still under development, could help surgeons of all levels to achieve better performances for ureteroscopic maneuvers compared with using fluoroscopic guidance. It also has the potential to reduce radiation exposure during fluoroscopy.

MP72-06 CORRELATION OF THE INSULIN RECEPTOR EXPRESSION AND THE RESISTANCE TO VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR TYROSINE KINASE INHIBITORS IN ADVANCED CLEAR CELL RENAL CELL CARCINOMA

INTRODUCTION AND OBJECTIVES: Vascular endothelial growth factor receptor tyrosine kinase inhibitors (VEGFR-TKIs) demonstrate the significant efficacy for advanced clear cell renal cell carcinoma (ccRCC), however, it eventually becomes resistant to VEGFR-TKIs during the treatment. So far, the mechanisms for the resistance to VEGFR-TKIs remain to be fully elucidated. Previously we have identified the gene set which could predict poor prognosis of ccRCC patients (Takahashi M et.al., Proc Natl Acad Sci U S A., 98: 9754, 2001). We examined whether the insulin receptor (INSR) expression in the gene set may correlate with the resistance to VEGFR-TKIs. METHODS: The INSR expression was examined immunohistochemically in the nephrectomy specimens of the RCC patients (n1⁄433) who then received axitinib as the VEGFR-TKI and correlated with their survival outcome. We compared the INSR expression of the nephrectomy or metastasectomy specimens before and after the administration of VEGFR-TKIs in 5 cases. The INSR expression of the human renal glomerular endothelial cells (HGEC) was compared before and after the administration of axitinib by Western blotting. In addition, we established patient derived Xenograft model (PDX) of ccRCC. Tumors of PDX were resected when it regrew and showed the resistance for axitinib and the INSR expression was compared before and after the treatment by Western blotting. RESULTS: The INSR was expressed at the vessels surrounding tumor cells. Progression-free survival (PFS) was significantly shorter in the INSR-negative group than in the INSR-positive group. Multivariate analysis revealed that the INSR expression was the significantly independent predictor of PFS. In the specimens resected after VEGFR-TKI, the INSR expression was more frequently decreased. As the concentration of axitinib increased, the INSR expression in the HGEC was decreased. The tumors of PDX that were resected after demonstrating the resistance to axitinib had the decreased INSR expression. CONCLUSIONS: The decreased INSR expression could be correlated with the resistance to VEGFR-TKI and its expression may be useful in selecting appropriate drugs for advanced ccRCC patients. Source of Funding: None

MP45-17 GALECTIN-3 PLAYS CRITICAL ROLES FOR THE GROWTH OF BENIGN PROSTATIC HYPERPLASIA

INTRODUCTION AND OBJECTIVES: Galectin-3, a multifunctional oncogenic protein, has been reported to play important roles of progression in a variety of cancer including prostate cancer through the regulation of cancer cell proliferation, apoptosis, invasion and metastasis. However, we also identified the frequent up-regulation of Galectin-3 in benign prostatic hyperplasia (BPH), yet its pathophysiological roles in BPH. Here, we report the involvement of Galectin-3 in the growth of BPH. METHODS: To investigate the association of Galectin-3 expression and prostate volume, we examined serum Galectin-3 (pg/ ml) with ELISA method in non-cancer cohort and analyzed the correlation between Galectin-3 expression and prostate volume with Speamans correlation coefficient. Next, to analyzed proliferation abilities and the effect of Galectin-3 on smooth muscle, we examined knockdown of Galectin-3 expression by siRNA in BPH1 cells (benign prostatic hyperplasia cell line) and a co-culture experiment of BPH1 and PrSMC (Normal Human Prostate Smooth Muscle. Cells). Moreover, to investigate biological function of Galectin-3, we examined the gene expression profiles in Galectin-3-depleted BPH1 cells with microarray and bioinformatics analyses. RESULTS: Correlation analysis revealed that serum Galectin-3 (ng/ml) were correlated with prostate volume (R1⁄40.643 p1⁄40.023) (Figure 1). Next, depletion of Galectin-3 suppressed cell proliferation in BPH1 cells. Moreover, depletion of Galectin-3 in BPH1 cells suppressed cell proliferation of PrSMC cells in a co-culture method, suggesting Galectin-3 enhanced proliferation of PrSMC cells. Bioinformatics analysis with GSEA revealed that depletion of Galectin-3 was involved in interferon a response and interferon a response and interferon ? response (FDR q value < 0.001) (Figure 2), suggesting Galectin-3 regulates the proliferation of PrSMC through interferon response. CONCLUSIONS: Our findings suggest that Galectin-3 is significantly involved in the growth of BPH and a promising therapeutic target for patients with BPH. Source of Funding: GSK Japan Research Grant 2016

Substitution of anti-androgens and tegafur-uracil combination therapy for castration-resistant prostate cancer: Results of a multi-center randomized phase II study

We conducted this study to determine whether substitution with anti-androgen (SOA) and tegafur-uracil (a pro‑drug of 5-FU) combination therapy is more effective than SOA alone after relapse from initial hormonal therapy. Patients who were histologically confirmed and relapsed after initial hormonal therapy were included. All patients were randomly allocated into two groups: SOA alone (group A) or SOA combined with tegafur-uracil (group B). The mRNA expression of four enzymes, including thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), orotate phospho-ribosyltransferase (OPRT) and thymidine phosphorylase (TP), in prostate cancer cells was analyzed by quantitative reverse-transcription polymerase chain reaction. Fifty-two patients were enrolled in this study. The median age was 77 (range: 47-92) years. The PSA response rate in group B (61.5%) tended to be higher compared to that in group A (34.6%) (p=0.095). Group B (median: 15.9 months) had a significantly longer time to PSA progression (TTP) compared to group A (6.4 months) (p=0.014). In patients with a lower TS expression or a higher OPRT expression, group B demonstrated a higher PSA response rate compared to group A (p=0.019 and p=0.041, respectively). In addition, in the patients with a lower TS expression, group B demonstrated a significantly longer TTP compared to group A (p=0.018). There were no severe adverse events in either treatment group. After relapse from initial hormonal therapy, SOA combined with tegafur-uracil is effective and well tolerated. The TS mRNA expression level may be a predictive factor for this combination therapy.