Sadao Kamidono

Conventional Versus Microdissection Testicular Sperm Extraction for Nonobstructive Azoospermia

PURPOSE We established a practical and safe strategy for testicular sperm extraction (TESE) in patients with nonobstructive azoospermia and compared conventional with microdissection TESE. MATERIALS AND METHODS In a retrospective comparative study 46 patients, including 22 with obstructive and 24 with nonobstructive azoospermia, underwent conventional TESE. Another 100 patients, including 26 with obstructive and 74 with nonobstructive azoospermia, underwent microdissection TESE. Conventional TESE was performed via 3 small 5 mm. incisions in the tunica albuginea. Microdissection TESE was performed by making a 3 to 4 cm. incision in the tunica albuginea under operating microscopy, avoiding the underlying testicular artery. Seminiferous tubules that appeared dilated and opaque were harvested. Sperm recovery rates were compared, as were complication rates assessed by ultrasonographic and endocrinological evaluations. RESULTS In obstructive azoospermia cases the sperm recovery rate was 100% for each procedure. In nonobstructive azoospermia cases sperm were recovered in 16.7% and 44.6% by conventional and microdissection TESE, respectively (p = 0.0271). In cases of histologically diagnosed maturation arrest the sperm recovery rate was 37.5% and 75%, respectively (p = 0.22585). In cases of the Sertoli-cell-only syndrome the sperm recovery rate was 6.3% and 33.9%, respectively (p = 0.0494). We identified dilated and opaque seminiferous tubules containing spermatozoa under operating microscopy in 22.2% of patients with maturation arrest and in 63.2% with the Sertoli-cell-only syndrome. The complication rate was significantly lower for microdissection than for conventional TESE. CONCLUSIONS In nonobstructive cases, especially those of the Sertoli-cell-only syndrome, microdissection TESE can effectively retrieve spermatozoa and minimize the risk of complications.

Elevation of serum levels of urokinase-type plasminogen activator and its receptor is associated with disease progression and prognosis in patients with prostate cancer.

Several investigators have revealed that urokinase‐type plasminogen activator (uPA) and its receptor (uPAR) are overexpressed in serum as well as in tumor tissues in patients with various types of cancer. In this study, we examined whether the serum levels of uPA and uPAR could be used as predictors of the progression and prognosis of prostate cancer.

Serum matrix metalloproteinase-2 and its density in men with prostate cancer as a new predictor of disease extension.

We examined whether the serum matrix metalloproteinase‐2 (MMP‐2) level and MMP‐2 density could be predictors of the development and extension of prostate cancer. Serum samples were collected before any clinical treatment from 98 patients with prostate cancer and from 76 patients with benign prostatic hyperplasia (BPH). Control sera were obtained from 70 healthy men. The serum level of MMP‐2 was determined by 1‐step enzyme immunoassay. A newly defined MMP‐2 density parameter was determined by dividing the serum level of MMP‐2 by the prostate volume, which was measured by ultrasonography. The mean serum level of MMP‐2 in prostate cancer patients was significantly higher than in the control and BPH groups. Furthermore, the serum MMP‐2 levels in prostate cancer patients with metastasis were highly elevated compared with those without metastases. The MMP‐2 density in pathologically organ‐confined prostate cancer was significantly higher than that in BPH. There was a statistically significant difference in the MMP‐2 density between pT2N0M0 and pT1N0M0 prostate cancers. Moreover, the serum MMP‐2 level correlated well with the clinical course of prostate cancer with bone metastasis. Our results suggest that MMP‐2 plays an important role in the development and extension of prostate cancer and that the serum level of MMP‐2 and the MMP‐2 density indicate prostate cancer extension and are, therefore, useful for the followup of prostate cancer patients. Int. J. Cancer (Pred. Oncol.) 79:96–101, 1998.

Overexpression of Bcl-2 in bladder cancer cells inhibits apoptosis induced by cisplatin and adenoviral-mediated p53 gene transfer

To investigate the effects of the expression of Bcl-2 protein in bladder cancer on the apoptosis induced by cisplatin or adenoviral-mediated p53 gene (Ad5CMV-p53) transfer, we transfected the bcl-2 gene into KoTCC-1, a human bladder cancer cell line that does not express the Bcl-2 protein. The Bcl-2-transfected KoTCC-1 (KoTCC-1/B) exhibited significantly higher resistance to both cisplatin and Ad5CMV-p53 transfer than did either the parental KoTCC-1 (KoTCC-1/P) or the vector-only transfected cell line (KoTCC-1/C). The flow cytometric analysis of the propidium iodide-stained nuclei and DNA fragmentation analysis after cisplatin or Ad5CMV-p53 treatment revealed DNA degradation in both KoTCC-1/P and KoTCC-1/C, whereas KoTCC1/B showed a marked inhibition of DNA degradation. Following the treatment with cisplatin or Ad5CMV-p53, the accumulation of p53 protein was highly detectable for a long period in KoTCC-1/B compared to that in KoTTC-1/P and KoTCC-1/C. Furthermore, the cisplatin and Ad5CMV-p53 treatments each reduced the volume of the subcutaneous tumors established in nude mice formed by KoTCC-1/P or KoTCC-1/C; in contrast, their reductive effects on the tumors formed by KoTCC-1/B were significantly suppressed. The intraperitoneal tumor cell implantation model revealed that the prognoses of mice injected with KoTCC-1/B were significantly inferior to those of the mice injected with either KoTCC-1/P or KoTCC-1/C after treatment with cisplatin or Ad5CMV-p53. These findings suggest that the expression of Bcl-2 in bladder cancer cells interferes with the therapeutic effects of cisplatin and Ad5CMV-p53 through the inhibition of the apoptotic pathway.

Overexpression of clusterin in transitional cell carcinoma of the bladder is related to disease progression and recurrence.

OBJECTIVES To determine whether the expression level of clusterin in transitional cell carcinoma (TCC) of the bladder could be used as a predictor of tumor recurrence and prognosis. METHODS Total RNA samples were extracted from 89 specimens of TCC of the bladder, and the expression level of clusterin mRNA in these specimens was measured by Northern blot analysis. The results were evaluated with respect to several clinicopathologic factors. RESULTS The mean level of clusterin mRNA expression in invasive TCC of the bladder was fourfold and fivefold higher than that in superficial TCC and normal urothelial tissue, respectively. The expression level of clusterin mRNA showed no significant correlation with sex, age, tumor size, or multiplicity, and the pathologic stage and tumor grade showed close associations with clusterin expression. The overall survival rate of patients with strong clusterin expression was significantly lower than that of patients with weak expression. Among the 43 patients with invasive TCC who underwent complete resection, the recurrence-free survival rate of patients with strong clusterin expression was significantly lower than that of patients with weak expression. Moreover, multivariate analyses indicated that among these 43 patients, strong expression of clusterin was an independent predictor of tumor recurrence. CONCLUSIONS. These findings indicate that clusterin mRNA is strongly expressed in invasive TCC of the bladder compared with the expression in superficial TCC and that strong clusterin expression could be used as a novel predictor of prognosis of patients with TCC of the bladder.

Stress protein GRP78 prevents apoptosis induced by calcium ionophore, ionomycin, but not by glycosylation inhibitor, tunicamycin, in human prostate cancer cells

GRP78 induction has recently been shown to play a critical role in maintaining cell viability against several kinds of stress, including depletion of endoplasmic reticulum Ca2+ and accumulation of unglycosylated proteins, under specific experimental conditions. However, the functional significance of GRP78 induction after stressful treatment has not been well defined. This article characterizes the different biological features associated with GRP78 induction by two kinds of stress agents, calcium ionophore, ionomycin (IM), and glycosylation inhibitor, tunicamycin (TM), focusing on the association with apoptosis in human prostate cancer cells. Both IM and TM treatment resulted in marked induction of GRP78 transcription in androgen‐dependent prostate cancer LNCaP cells maintained in medium without androgen, but not in medium containing androgen, as measured by Northern blotting and nuclear run‐off assays. After pretreatment with tumor necrosis factor‐α, which has potent cytotoxic effects on LNCaP cells, both IM and TM could induce substantial increases in GRP78 transcription in LNCaP cells, even in medium containing androgen. Under both experimental conditions described, DNA fragmentation assays showed a direct correlation between the onset of apoptosis in LNCaP cells after IM treatment and the initiation of GRP78 transcript induction, while induction of GRP78 expression preceded TM‐induced apoptosis. To elucidate the functional differences of GRP78 induction by IM and TM, an antisense oligodeoxynucleotide (ODN) targeted against the grp78 gene was designed to reduce GRP78 expression in a sequence‐specific and dose‐dependent manner. Antisense GRP78 ODN treatment substantially enhanced apoptosis of LNCaP cells induced by IM compared with mismatch control ODN treatment, whereas no marked differences were observed in apoptotic features induced by TM with antisense GRP78 and mismatch control ODN treatment. Studies of additional androgen‐independent prostate cancer PC3 cells also demonstrated a correlation between GRP78 induction and resistance to apoptosis after IM treatment, but not after TM treatment. These findings suggest that there are at least two GRP78 signaling pathways, which play different roles in resistance against stress‐induced apoptosis. J. Cell. Biochem. 77:396–408, 2000.

Quality of life of living kidney donors: the short-form 36-item health questionnaire survey.

OBJECTIVES To determine the psychological and social effects of kidney donation on kidney donors by using the short-form 36-item health survey (SF-36) as the quality-of-life questionnaire. METHODS A total of 104 living donor nephrectomies have been performed at Kobe University Hospital and Nishinomiya Prefectural Hospital. We mailed the questionnaires to donors or handed them out directly at the outpatient clinic. The first part of the questionnaire consisted of the SF-36 (limitations on physical functioning because of health problems) and the second part consisted of 15 questions about donation-related stress, expenses incurred, physical changes, and pre-existing factors such as relationship to the recipients. RESULTS The SF-36 and the questionnaire about donor satisfaction were completed by 69 donors (48 women and 21 men; mean age 52.1 +/- 8.2 years), only 6 of whom (9%) reported minor complications with the donor operation. The SF-36 scores of our donors were not significantly different from that of the general U.S. population and U.S. donors. In some categories (physical functioning, role-physical, bodily pain, general health, vitality, and mental health), our donors scored slightly higher than the U.S. general population. Although 97% of the donors would make the same choice again, 3% believed that donating had had a negative impact on their health, and 16% reported negative financial consequences. CONCLUSIONS The quality of life for kidney donors was not affected by donor nephrectomy. Living kidney transplantation seems to be suitable for the rescue of patients with end-stage renal disease. Better psychological and technical preparation for surgery and more consistent follow-up may reduce the negative outcomes even further.

Basic fibroblast growth factor regulates matrix metalloproteinases production and in vitro invasiveness in human bladder cancer cell lines.

PURPOSE This study was designed to investigate the effect of endogenous basic fibroblast growth factor (FGF-2) on matrix metalloproteinases (MMPs) production and in vitro invasive potential of human bladder cancer cell lines. MATERIALS AND METHODS The human bladder cancer cell lines, HT1376 and KoTCC-1, were used in this study. The mRNA for FGF receptor has been shown to be expressed in both cell lines; the mRNA for FGF-2 is expressed in only KoTCC-1. The effects of FGF-2 expression on HT1376 by gene transfection and those of FGF-2 antisense oligonucleotides treatment on KoTCC-1 were analyzed by zymography and in vitro tumor cell invasion assay. RESULTS The introduction of human FGF-2 gene into HT1376 cells markedly enhanced both the MMP-2 and MMP-9 production, and the in vitro invasive potential was also increased. In contrast, the exposure of KoTCC-1 cells to FGF-2 specific antisense oligonucleotides decreased the MMP-2 production and in vitro invasive potential, but the exposure to FGF-2 sense oligonucleotides did not. CONCLUSIONS These findings suggest that FGF-2 plays an important role in the invasive process of human bladder cancer in part through the regulation of MMPs production.

Increased angiogenin expression in the tumor tissue and serum of urothelial carcinoma patients is related to disease progression and recurrence.

The progression of solid tumors is at least partly dependent on angiogenesis, the induction of which is mediated by several angiogenic factors, including angiogenin (ANG). The authors evaluated the expression of ANG in the tumor tissue and serum of patients with urothelial carcinoma.

Health-Related quality of life with orthotopic neobladder versus ileal conduit according to the SF-36 survey

OBJECTIVES To compare health-related quality of life (HRQOL) in patients with a neobladder and in patients with an ileal conduit. METHODS HRQOL was assessed using the SF-36 survey, supplemented with a questionnaire concerning micturition status. RESULTS Patients (n = 36) with a neobladder were significantly younger at time of surgery and time of survey than patients with an ileal conduit (n = 20). Mean (+/- SD) follow-up periods for patients with a neobladder and with an ileal conduit were 31.3 +/- 33.1 and 44.8 +/- 30.7 months, respectively. No significant difference was apparent in any scale score between neobladder and ileal conduit groups. Role-physical functioning (RP) and role-emotional functioning (RE) scale scores in both neobladder and ileal conduit patients appeared to be below the general U.S. population norm. Patients with neobladder 65 years old or older (n = 17) showed significantly lower RP and RE scores than those younger than 65 years (n = 19; P <0.05). Duration of follow-up was not related to scale scores. Continence status did not measurably affect HRQOL. CONCLUSIONS All scales concerning HRQOL except RP and RE were favorable with both neobladder and ileal conduit, and no significant differences were observed between these two types of urinary tract reconstruction. Generally, patients with a neobladder or an ileal conduit were satisfied with their overall quality of life and health.