Akihiko Hatano

Expression of m2 muscarinic acetylcholine receptor mRNA in primary culture of human prostate stromal cells.

Abstract The aim of this study was to investigate the expression of the muscarinic acetylcholine receptor (mAchR) subtypes mRNA in primary culture of human prostate stromal cells using the reverse transcription polymerase chain reaction (RT-PCR), RNA blotting and in situ hybridization (ISH). Using an explant method, we obtained a primary culture of prostate stromal cells from three patients with benign prostatic hypertrophy. Total RNA was extracted using the acid guanidinium method for cDNA synthesis. First-strand cDNA was then used for PCR with primers designed to amplify the fragments of each mAchR subtypes (m1–m5) cDNA sequence. The m2, m3 and m4 subtype expected bands were detected; in particular m2 transcripts was strongly detected in the stromal cell culture. Each of the PCR products were subcloned into the pGEM-T plasmid vector, sequenced and random primer labeled using 32P. Digoxigenin-labeled cRNA probes were synthesized by in vitro transcription. RNA blotting using a m2 muscarinic receptor cDNA probe revealed a 4.5 kb single transcript. However, m3 and m4 probes did not hybridize. Using in situ hybridization (ISH), m2 receptor mRNA signals were detected in several smooth muscle cells. The staining was predominantly localized to the perinuclear cytoplasm. The m3 and m4 probes did not hybridize. These results suggested that m2 receptor subtype plays a role in smooth muscle activity of the human prostate.

Changes in urinary excretion of endothelin-1-like immunoreactivity before and after unilateral nephrectomy in humans. Comparison with other urinary parameters and unilateral adrenalectomy.

To assess the value of endothelin-1 (ET-1) as a parameter of renal functional overload and the process of hyperfiltration, urinary excretion of ET-1-like immunoreactivity (U-ET-1) and urinary excretion of other parameters [beta 2-microglobulin (beta 2-MG), N-acetyl-beta-D-glucosaminidase (NAG), albumin (Alb) and 24-hour creatinine clearance (Ccr24)] were measured before, 1, 2 and 7 days after unilateral nephrectomy in 15 patients with unilateral renal lesions (renal cell cancer, renal pelvic cancer, ureteral cancer and renal tuberculosis) and after unilateral adrenalectomy in 5 patients with adrenal lesions. In the nephrectomy group (NX), Ccr24 significantly decreased at 1 and 2 days after the operation, but the other 4 parameters significantly increased after the operation as compared to preoperative values. In the adrenalectomy group (ADX), neither Ccr24 nor ET-1/Cr showed any change after the operation, but the other 3 parameters (beta 2-MG/Cr, NAG/Cr and Alb/Cr) significantly increased. Comparing the NX and ADX groups, both ET-1/Cr and beta 2-MG/Cr in NX were significantly higher than in ADX after the operation. On the other hand, neither NAG/Cr nor Alb/Cr in NX differed from values in ADX. Comparing ET-1/Cr and other parameters, only beta 2-MG showed a positive significant correlation. These results suggest that U-ET-1 may be an indicator of functional overload of the kidney.

Alpha-1 adrenoceptor subtypes (high, low) in human benign prostatic hypertrophy tissue according to the affinities for prazosin

A novel classification of α‐1 adrenoceptor subtypes (High, Low) was applied to human benign prostatic hypertrophy (BPH)tissue.

Value of Urinary Endothelin-1 in Patients with Primary Vesicoureteral Reflux

By using a radioimmunoassay specific for endothelin-1 (ET-1), we measured urinary excretion of ET-1-like immunoreactivity (LI) in 63 spot urine samples of 48 patients with primary vesicoureteral reflux (VUR). And also, urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG), beta 2-microglobulin (beta-2-MG), microalbumin (Alb) and creatinine (Cr) were measured. There was no significant correlation in any of the pairs ET-1 and NAG, ET-1 and beta 2-MG, and ET-1 and Alb. Comparing the grade of reflux according to the International Classification with urinary ET-1, urinary ET-1/Cr levels in patients with grade 2, 3 and 4 VUR were higher than normal, and the ratio of more than normal urinary ET-1/Cr increased in proportion to the grade of reflux, but it conversely decreased in grade 5. In conclusion, urinary ET-1 may be an indicator of renal tubular injury in patients with primary VUR, and its meaning may be different from conventional urinary parameters.

α1- and α2-Adrenoceptors in BPH

The dynamic obstruction of the bladder outlet secondary to benign prostatic hyperplasia (BPH), and the contractile properties of the human prostate are mediated primarily by α₁-adrenoceptors. There are now at least three subtypes (A, B, and D) of α₁-adrenoceptors, and recent work revealed that α_1A-adrenoceptor and α_1B-adrenoceptor may have a prime role for prostatic obstruction, and contraction of artery, respectively. Very recently, the presence of a low affinity α₁-adrenoceptor for prazosin, named α_1L, in the human BPH tissue has been determined. Because the DNA sequence of α_1L-adrenoceptor has not yet been cloned, the α_1L-adrenoceptor may be another form of the α_1A-adrenoceptor, or another pharmacologically distinct α₁-adrenoceptor which mediates the norepinephrine-induced contraction of the prostatic smooth muscle. Furthermore, the contribution of α₁-adrenoceptors in the prostate to symptoms (not only obstructive, but irritative symptoms) which are elicited by prostatic obstruction remains to be determined.

Changes in urinary excretion of endothelin-1-like immunoreactivity in patients with testicular cancer receiving high-dose cisplatin therapy.

To assess the value of endothelin-1 (ET-1) in estimating renal injury in patients receiving high doses of cisplatin, urinary excretion of ET-1-like immunoreactivity (U-ET-1), beta 2-microglobulin (U-beta 2-MG), and N-acetyl-beta-d-glucosaminidase (NAG) were measured before, 1 week after, and 2 weeks after the administration of cisplatin in eight patients with testicular cancer (mean age, 33.3 years). Levels of U-ET-1/creatinine (Cr) during and 1 week after cisplatin treatment were significantly higher than before cisplatin treatment. There were no differences in U-ET-1/Cr levels during, 1 week after, and 2 weeks after cisplatin treatment. The level of U-beta 2-MG/Cr during cisplatin treatment was significantly higher than levels before, 1 week after, and 2 weeks after treatment. However, there were no differences in U-beta 2-MG/Cr levels before, 1 week after, and 2 weeks after cisplatin treatment. The level of U-NAG/Cr during cisplatin treatment was higher than levels before, 1 week after, and 2 weeks after treatment; U-NAG/Cr during cisplatin treatment was higher than levels before, 1 week after, and 2 weeks after treatment; U-NAG/Cr gradually decreased after cisplatin treatment. Among the three parameters, only U-ET-1/Cr maintained a higher level after cisplatin treatment. The U-beta 2-MG/Cr level returned most rapidly to normal after cisplatin treatment. Although U-ET-1/Cr did not show any significant correlation with U-NAG/Cr (r = 0.282, P = NS), it showed a significant correlation with U-beta 2-MG/Cr (r = 0.454, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Application of flexible renoureteroscope for antegrade urethroscopy in the treatment of congenital anterior urethral valve

Antegrade urethroscopy was performed through a cystostomy using a 10.8-fr caliber flexible renoureteroscope in 4 children (aged 2 years and 8 months, 2 months, 14 years and 1 month) with anterior urethral valve. The anatomy of the valves as well as their destruction after transurethral resection (TUR) were well visualized by antegrade urethroscopy. This technique was helpful in performing TUR-valve, after which the findings of voiding cystourethrography in every case and the findings of uroflowmetry in the elder 2 cases showed remarkable improvement. This technique is relatively simple to perform and is thought to be one of the best methods to treat anterior urethral valve. This technique is also applicable to other lower urinary tract lesions in children.

Endothelin receptors in the human urinary bladder are different from those in the human ureter

Human urinary bladders from ten patients undergoing total cystectomy for bladder cancer and human ureters from ten patients undergoing radical nephrectomy for renal cell cancer were used. Strips (10 mm long, 1-2 mm wide) from the dome of urinary bladder and spirally cut strips (10 mm long, 2-4 mm wide) of ureter freed from mucosa were used. Informed consent was obtained from each patient before operation. Isometric contraction experiments were performed under 0.5 g tension in 10-ml organ baths containing modified KrebsHenseleit solution using indomethacin (1 gm). After 1 h equilibration and a reference contraction by 116.9 mM KC1, subsequent reponses were expressed as a percentage of the reference contractions obtained following completion of the experiment. Cumulative concentration-response curves to endothelin-1 (ET1, 0.1-0.1 gM) and sarafotoxin

Bladder Neck Suspension Using Percutaneous Bladder Neck Stabilization to the Pubic Bone with a Bone-Anchor Suture Fixation System:A New Extraperitoneal Laparoscopic Approach

6C [3] (

Measurement of urinary endothelin-1-like immunoreactivity and comparison with other urinary parameters in patients with primary vesicoureteral reflux : a preliminary report

6C; 0.1-0.1 gM) were constructed. The following six groups of experiments were performed on each sample. In groups 1, 3, 4, 5, and 6, a cumulative concentration-response curve was constructed for ET-1. In groups 3, 4, 5, and 6, BQ123Na [2] (10 nM, 0.1 gM, 1 I.tM, 10 gM) was added 30 min before administration of ET-1. In group 2, a cumulative concentration-response curve was constructed for