[Show abstract][Hide abstract] ABSTRACT: We investigated the ability of autologous adipose-derived cells injected into cryo-injured rabbit urethras to improve urinary continence and explored the possible mechanisms by which it occurred. Adipose tissue was harvested from the perivesical region of nine 10-week-old female New Zealand White rabbits and cultured for 7 days. Immediately after harvesting the tissue, we injured the internal urethral orifice by spraying liquid nitrogen for 20 sec. The cultured cells expressed the mesenchymal cell marker STRO1 but not muscle cell markers myoglobin or smooth muscle actin. Just prior to implantation, the adipose-derived cells were labeled with PKH26 fluorescent cell linker. Autologous 0.5×106 adipose-derived cells (5 rabbits) or cell-free control solution (4 rabbits) was injected around the cryo-injured urethras at 7 days after injury. Fourteen days later, leak point pressure was measured, and the urethras were harvested for immunohistochemical analyses. At 14 days after implantation, leak point pressure of the cell-implanted group was significantly higher compared to the cell-free control group (P<0.05). In immunohistochemical examination, the reconstructed skeletal and smooth muscle areas in the cell-implanted regions were significantly more developed than those in controls (P<0.05). Implanted PKH26-labeled adipose-derived cells were immunohistochemically positive for myoglobin, smooth muscle actin, and Pax7 antibodies, which are markers for skeletal muscle, smooth muscle, and myoblast progenitor cells, respectively. In addition, these implanted cells were positive for the nerve cell markers tubulin β3, S100, and the vascular endothelial cell marker von Willebrand factor. Furthermore, some of the implanted cells were positive for transforming growth factor β1, nerve growth factor, and vascular endothelial growth factor. In conclusion, implantation of autologous adipose-derived cells into the cryo-injured rabbit urethras promoted the recovery of urethral function by myogenic differentiation, neuroregeneration, and neoangiogenesis of the implanted cells and/or the surrounding tissues as well as by bulking effects. Thus, treatment of human radical prostatectomy-related stress urinary incontinence by adipose-derived cell implantation could have significant therapeutic effects.
Tissue Engineering Part A 02/2014; · 4.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objectives: Alpha-1 adrenoceptor (AR) antagonists are commonly used as therapeutic agents for patients with benign prostatic obstruction (BPO). Our objective was to investigate the correlation between the ratio of bladder mucosal alpha-1D/alpha-1A adrenoceptor mRNA and lower urinary tract function in BPO patients. Methods: In 20 BPO patients, the expression level of alpha-1 AR mRNAs in the bladder mucosal biopsies was investigated by reverse transcriptase polymerase chain reaction. The subjects were divided into two groups. In Group 1, the ratio of alpha-1D mRNA to alpha-1A mRNA was greater than one. In Group 2, the ratio was less than one. We determined the correlation by Schäfer nomogram between Group 1 and Group 2 patients and lower urinary tract function as determined by a video urodynamic study. Results: Two patients were excluded due to inability to void. Another was excluded because the alpha-1D/alpha-1A mRNA ratio was one. On the Schäfer nomogram, six of nine Group 1 cases had obstructions less than IV and normal or weak detrusor contractility. For Group 2, six of eight cases had obstructions more than IV and normal or strong detrusor contractility. Conclusion: Patients with higher levels of alpha-1D AR mRNA were distinct from those with higher alpha-1A AR mRNA levels with regard to obstruction and detrusor activity. The results suggest that the Schäfer nomogram might be useful in determining which alpha-1 AR antagonists are better for BPO patients suffering from storage symptoms.
[Show abstract][Hide abstract] ABSTRACT: Historically, α1-adrenoceptors have been classified into three subtypes (α1A, α1B and α1D) that are widely distributed in various organs. Research on the α1D-adrenoceptors in the bladder, urethra and prostate has focused on the relationship between expression levels and symptoms of bladder outlet obstruction, and the implications and functional roles of α1D-adrenoceptors subtypes in these organs. The α1D-adrenoceptor messenger ribonucleic acid and protein seem to be increased in obstructed bladders or small capacity bladders. In contrast, α1D-adrenoceptor subtype knock-out mice have been found to have a prolonged voiding interval. Interestingly, an α1D-adrenoceptor antagonist was found to inhibit the facilitation of afferent nerve activity for the micturition reflex induced by intravesical infusion of acetic acid. Clinically, patients who felt urgency at low filling volumes and had a small bladder capacity were found to have more α1D-adrenoceptor messenger ribonucleic acid in their bladder mucosa than patients who felt urgency at high filling volumes and had a large bladder capacity. An α1D-adrenoceptor antagonist was found to increase the first desired volume and the maximum desired volume while decreasing detrusor overactivity in pressure flow studies. Thus, α1D-adrenoceptors in the lower urinary tract might play an important role in the pathophysiology of lower urinary tract disorders.
International Journal of Urology 01/2013; 20(1). · 1.73 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: PURPOSE: To study if cold stress-induced detrusor overactivity in ovariectomized rats is associated with increased thermosensitive transient receptor potential melastatin 8 (TRPM8) channel expression in the skin, and if the response can be inhibited by alpha1-adrenergic receptor blockade. MATERIALS AND METHODS: Twenty-four Sprague-Dawley rats at postnatal week 30 were randomly selected for ovariectomy (OVX, n=16) or sham ovariectomy (n=8). Five weeks later, cystometric measurements of the conscious, free-moving rats were performed at room temperature (RT, 28 ± 2ºC) for 20 min. Eight OVX rats were intravenously administered 1.0 mg/kg naftopidil, and the other OVX and sham-operated rats (n=8 each) were given naftopidil-free vehicle. Five min later, they were transferred to a low temperature (LT) environment (4 ± 2ºC) and micturition patterns were again recorded. TRPM8 channel expression in lumbar skin was estimated by real-time reverse-transcriptase polymerase chain reaction and immunohistochemistry. RESULTS: TRPM8 channel mRNA and protein in the skin of OVX rats were significantly higher than in sham-operated rats. At RT, micturition parameters were similar for sham-operated and OVX rats. At LT, both sham-operated and OVX rats exhibited cold stress-induced detrusor overactivity, but the increased micturition frequency and decreased bladder capacity were significantly greater for OVX rats. Treatment of OVX rats with naftopidil inhibited the cold stress-induced detrusor overactivity. CONCLUSION: Cold stress-induced detrusor overactivity in rats with decreased estrogen is associated with an upregulation TRPM8 channels in the skin, and is mediated by nerve pathways utilizing alpha1-adrenergic receptors.
The Journal of urology 10/2012; · 4.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective: Cold stress can elicit increases in urinary urgency and frequency. We determined if there was a relationship between finger and toe temperatures and lower urinary tract symptoms (LUTS). Methods: We studied 50 people who visited a public health management seminar. The participants were divided into two groups according to self-described sensitivity to cold stress. The cold non-sensitive (CNS) group consisted of 3 males and 20 females (66.9 ± 10.8 years old), and the cold sensitive (CS) group consisted of 4 males and 23 females (65.8 ± 8.01 years old). Each participant was assessed to determine international prostate symptom score (IPSS), overactive bladder symptom score (OABSS), and quality of life (QOL) score. They were then instructed on lifestyle changes and exercises that could improve peripheral blood flow and provide relief for their LUTS. Next, the temperatures of their middle fingers and toes were measured before and after 5–10 min of the exercises. Two weeks later, the IPSS, OABSS, and QOL scores were reassessed. Results: Before exercise, the middle fingers were significantly warmer than the middle toes. Exercise had no significant effect on the middle finger temperature of either group; however, it did increase the middle toe temperature for both groups. The increase was greatest for the CS group. The CS group had higher LUTS storage symptoms than the CNS group, and these improved after 2 weeks of lifestyle changes and exercise. Conclusion: Improvements in lifestyle and daily exercise may be effective for LUTS in CS people.
[Show abstract][Hide abstract] ABSTRACT: We investigated expression of tachykinin peptides neurokinin A, neurokinin B, and substance P within urethras of spontaneously hypertensive rats (SHRs) and determined if a traditional Chinese herbal mixture, THC-002, decreased them.
Ten- and 40-week-old male SHRs were randomly separated into three groups (n = 12 each). Rats of one group were given orally 20 ml 0.9% NaCl solution per kg body weight daily for 1 week. One hour later, each received a similar volume of water. Rats in the second group were also given saline. One hour later, each received 20 mg THC-002 per kg body weight. The third group was untreated. The urethras were removed and separated into prostatic and non-prostatic regions and analyzed by real-time reverse transcription polymerase chain reaction (n = 6) and immunohistochemistry (n = 6).
In 40-week-old untreated SHRs, neurokinin B mRNA and protein, and substance P mRNA in prostatic urethras were significantly higher compared to the 10-week-old ones. Neurokinin A mRNA and protein, and substance P protein of the 40-week-old saline-loaded prostatic urethras were significantly higher compared to the 40-week-old untreated ones. In 40-week-old untreated SHRs, the non-prostatic urethral neurokinin B protein was significantly higher compared to the 10-week-old ones. In 40-week-old, saline-loaded SHRs, neurokinin A protein of the non-prostatic urethras was significantly higher compared to 40-week-old the untreated ones. In 40-week-old SHRs, THC-002 significantly decreased the expression of the urethral tachykinins, except for non-prostatic urethral neurokinin A mRNA.
Aging and saline-loading increased the expression of urethral tachykinin mRNAs and peptides. THC-002 partially decreased them.
Neurourology and Urodynamics 07/2011; 31(1):201-6. · 2.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective: Pressure-flow study is a method used to evaluate the degree of bladder outlet obstruction and the strength of detrusor contractility during voiding. However, whether or not the operation for benign prostate hyperplasia should be avoided in detrusor underactivity patients remains controversial. To address this, we performed a retrospective analysis of our pressure-flow study data for benign prostate hyperplasia patients. We especially focused on the backgrounds of patients with weak detrusor contractility.Methods: Patients (n = 288; average age, 71.5 years) who underwent pressure-flow study to evaluate operative indications between February 2001 and April 2010 were included in this study. We analyzed the relationships between background factors and detrusor contraction strength according to Schäfer's nomogram.Results: Patients with weak detrusor contractility had poor flow (5.81 mL/sec) and low voided volume (141.2 mL) compared to patients with normal (8.77 mL/sec, 202.0 mL) or strong (8.97 mL/sec, 178.3 mL) detrusor contractility. Twenty-six of 74 weak detrusor patients underwent prostate operation. The operated group had high obstruction grade (3.35, P < 0.001), but a low rate of detrusor overactivity (19.2%, P < 0.05), compared to the non-operated group (2.16, 41.7%). The operated group also had high urinary retention rate (38.5%) compared to the non-operation group (18.8%).Conclusion: We performed prostate surgery in patients who had episodes of urinary retention, with outlet obstruction, and with no detrusor overactivity, even in those with weak detrusor contractility. The operation may not be contraindicated for these patients. Pressure-flow study is an important tool to ensure adequate informed consent.
[Show abstract][Hide abstract] ABSTRACT: The purpose of this study was to determine if implantation of autologous bone-marrow-derived cells has the potential to treat stress urinary incontinence caused by intrinsic sphincter deficiency. Bone marrow cells harvested from femurs of New Zealand White rabbits were cultured for 10 days. Seven days before implantation, the urethral sphincters located at the internal urethral orifice were cryo-injured by spraying liquid nitrogen for 15 s. The cultured autologous bone-marrow-derived cells were implanted 7 days after cryo-injury. For controls, cell-free solutions were injected. At 7 and 14 days after implantation, leak point pressures were determined and the urethral sphincters were examined by immunohistochemistry. At 7 and 14 days, the cell-implanted regions contained numerous striated and smooth muscle-like cells expressing myoglobin and smooth muscle actin, respectively. The proportions of myoglobin- and smooth muscle actin-expressing areas in both the 7- and 14-day cell-implanted regions were significantly higher than in controls. By 14 days, these differentiated cells formed contacts with similar cells, creating layered muscle structures. At that time, the leak point pressure of the cell-implanted rabbits was significantly higher than that of the controls. In conclusion, autologous bone-marrow-derived cells can reconstruct functional urethral sphincters.
Tissue Engineering Part A 01/2011; 17(7-8):1069-81. · 4.64 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the possible relationship between the bladder mucosal expression of α(1)-adrenoceptor (AR) mRNAs, storage symptoms and urodynamic findings in patients with lower urinary symptoms (LUTS) and benign prostatic obstruction (BPO).
Mucosa was obtained from the posterior bladder wall from 20 patients with a diagnosis of LUTS and BPO undergoing prostatectomy. α(1)-AR subtype mRNA was quantified using competitive real-time reverse transcription-polymerase chain reaction. A preoperative video urodynamic study measured volumes for first desire to void (FDV) and strong desire to void (SDV). Patients were divided into group 1, with volumes for FDV ≤ 200 ml or SDV ≤ 300 ml, and group 2, with volumes FDV ≥ 201 ml and SDV ≥ 301 ml. Preoperative and postoperative international prostate symptom score (IPSS) and Overactive Bladder Symptom Score (OABSS) were evaluated.
The group 1 FDV (141.0 ± 13.5 ml) was significantly lower than that of group 2 (299.0 ± 48.1 ml). Similarly, the SDV for group 1 (247.0 ± 21.9 ml) was also significantly lower than for group 2 (444.0 ± 32.5 ml). There were no significant differences between groups 1 and 2 with respect to mucosal α(1a)- and α(1b)-AR mRNA levels. However, group 1 patients had significantly more α(1d)-mRNA than those in group 2. No relationship between the expression of α(1)-AR mRNAs and preoperative or postoperative OABSS or IPSS was detected.
There was a relationship between the expression of α(1d)-AR mRNA in the bladder mucosa and storage-phase urodynamics in LUTS/BPO patients, suggesting a role of α(1D)-ARs in bladder sensation.
Scandinavian Journal of Urology and Nephrology 10/2010; 45(1):15-9. · 1.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The Transient Receptor Potential Melastatin (TRPM)8 channel, activated by menthol and cool temperatures, has been proposed to be a cool receptor involved in mediation of the bladder response in the diagnostic ice water test. We examined the expression of TRPM8 in the rat skin and investigated if stimulation of skin menthol receptors by menthol can induce detrusor activity in conscious rats.
Immunofluorescence staining for TRPM8 was performed in samples of the skin from the leg and back of the rats. Continuous cystometry was performed in conscious animals. Voiding interval (VI), micturition volume (MV), and bladder capacity (BC) were evaluated before and after spraying menthol solution to the skin of the leg and back of the rats.
Immunohistochemical analysis revealed that TRPM8 was expressed in the investigated tissues. The expression of TRPM8-positive receptors was not significantly different between the leg and back skin. Saline caused no significant change in cystometric parameters. After spraying menthol solution (50% and 99% to the skin of the leg, and 99% to the back skin), VI, MV, and BC decreased significantly.
TRPM8 is expressed in rat skin. Spraying menthol solution onto the skin-induced detrusor activity, an effect we suggest is mediated by stimulation of TRPM8 receptors.
Neurourology and Urodynamics 08/2009; 29(3):506-11. · 2.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate possible mechanisms of action of THC-002 (HARNCARE), a galenical produced from the traditional Chinese herbal mixture Ba-Wei-Die-Huang-Wan, which has been reported to improve lower urinary tract symptoms (LUTS) in patients.
Forty-five female SHRs were randomly separated into three groups. Two groups were given 20 ml physiological saline solution (PSS) per kg-body weight orally daily for 1 week. An hour after the administration of PSS, one of the groups received 20 mg THC-002 per kg body weight, and the other a similar volume of THC-002-free saline. The third group received no treatments. The bladders were analyzed by real time RT-PCR (n = 6) and immunohistochemistry (n = 3) for the expression of tachykinins and P2X3 and TRPV1 receptors. Cystometric investigation (n = 6) was conducted after intravesical instillation of saline followed by 5 mg/ml ATP solution.
Treatment with PSS caused and upregulation of tachykinins and P2X3 and TRPV1 receptors, which was prevented in the group treated with THC-002. In the normal (non-treated) and non-THC-002-treated SHRs, instillation of the ATP solution decreased voiding interval, micturition volume, and bladder capacity compared to the instillation of saline. However, in the THC-002-treated SHRs, ATP instillation had no effect.
In SHRs, THC-002 reduced the bladder expression of tachykinins and P2X3 and TRPV1 receptors, and inhibited ATP-induced detrusor overactivity. These effects may explain part of its beneficial effects on LUTS.
Neurourology and Urodynamics 04/2009; 28(6):529-34. · 2.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A 17-year-old woman was admitted to our hospital for the investigation of asymptomatic gross hematuria. Twenty nine months ago, she had received the right lower lobectomy because of the malignant salivary gland type mixed tumor of the bronchus. The intravenous pyelography and the right retrograde pyelography showed the irregular wall of the right renal pelvis. The computerized tomography showed a hypodense tumor which occupied the lower half of the right kidney. Cystoscopy showed bleeding from the right ureteral orifice. Chest X-ray revealed multiple lung metastases at the first examination, but right nephrectomy was performed because gross hematuria continued. The tumor thrombus, which spreaded into vena cava, was removed. The pathological diagnosis was the metastatic renal tumor of malignant salivary gland type mixed tumor. The patient died 102 days postoperatively.
Nippon Hinyōkika Gakkai zasshi. The japanese journal of urology 09/1998; 89(8):734-7.
[Show abstract][Hide abstract] ABSTRACT: A 79-year-old woman was admitted to our hospital with gross hematuria and abdominal pain. She had had a uterine cancer 11 years previously and received 56 Gy 60Co external irradiation combined with 129 Gy 137Cs internal irradiation. She had a sign of pan-peritonitis. An emergency operation revealed an intraperitoneal rupture of the dome of the urinary bladder 8 cm in length. Because a primary suturing of the bladder wall was unsuccessful, bilateral cutaneous ureterostomy was performed. Histologically, the ruptured bladder wall showed a mucosal erosion and fibrosis of the muscle layer.