Transplant Procurement Management Model Training: Marked Improvement in the Mindset of In-Hospital Procurement Coordinators at Hyogo Prefecture, Japan.

BACKGROUND: The organ donation rate in Japan is much lower than that in other developed countries for several reasons. An advanced educational program for in-hospital procurement coordinators is a possible solution for this. We introduced a Transplant Procurement Management (TPM) educational program at Hyogo Prefecture, Japan. METHODS: Ten healthcare professionals at Hyogo Prefecture participated in the Advanced International TPM course to educate themselves on TPM and held 2 TPM Model Organ Procurement Training Workshops at Hyogo Prefecture for in-hospital procurement coordinators. Furthermore, we held 2 workshops outside Hyogo Prefecture and at the same time undertook a pre-workshop questionnaire survey to evaluate the ability and motivation with respect to organ donation. To evaluate the effectiveness of the workshops, we conducted post-workshop and 3-months-after workshop questionnaire surveys. RESULTS: The results of the pre-workshop survey revealed that in-hospital procurement coordinators lacked the knowledge regarding the entire organ donation process, the current status of organ donation in Japan, and the definition of brain death. Moreover, they did not completely understand the meaning of "organ donation." The results of the post-workshop questionnaire survey showed that the educational program was effective to improve the knowledge and skills of organ donation and motivated behavioral changes among the participants. CONCLUSIONS: The survey results showed that our TPM model educational program offered sufficient knowledge and skills to increase organ donation at Hyogo Prefecture. We will continue this program and make an effort to further contribute to the Japanese organ donation activities.

Rapid detection and differentiation of Gram-negative and Gram-positive pathogenic bacteria in urine using TaqMan probe.

Urinary tract infection has been shown to be quite complicated and often difficult to diagnose and treat. For appropriate diagnosis, it is very important to find the correct Gram stain classification as soon as possible, especially in severe cases where there is a possibility of severe sepsis developing. In order to solve this problem, we developed a new method to detect a Gram stain of bacteria obtained from 1 ml of urine from urinary tract infection patients using a consensus real-time PCR protocol with a TaqMan probe that allows detection of spiked bacterial 16S DNA from urine. We extracted DNA of 55 urine samples obtained from patients with complicated urinary tract infection and at the same time performed urine culture testing. After DNA extraction, they were subjected to real-time PCR using a TaqMan discrimination system. Sixteen kinds of bacteria were cultured from the urine culture testing. Of these bacteria, eight were classified as Gram-positive bacteria and the other eight were classified as Gram-negative bacteria. Of the 55 samples, the TaqMan technique result showed 27 samples that were classified as Gram-negative bacteria; 11 samples that were Gram-positive, 10 that included both Gram-negative and -positive bacteria, and 7 that showed no amplification. The classifications of all samples corresponded exactly to those determined by urine culture testing. The present genotyping method of real-time PCR using a TaqMan discrimination system could be applied to the rapid detection of Gram-positive or -negative bacteria in urine of urinary tract infection patients. This assay can differentiate those species tested, but whether the presence of other (untested) bacteria could lead to misinterpretation is unknown. For further investigation, it is important to test other (untested) bacteria in the near future.

Identification of parathyroid hormone-related protein-derived peptides immunogenic in human histocompatibility leukocyte antigen-A24+ prostate cancer patients.

Parathyroid hormone-related protein (PTHrP) is a key factor in the development of bone metastases, which are a major barrier in treating prostate cancer patients. In this study, we attempted to identify PTHrP-derived peptides immunogenic in human histocompatibility leukocyte antigen (HLA)-A24(+) prostate cancer patients. Among four different PTHrP peptides carrying the HLA-A24 binding motif, both the PTHrP(36-44) and PTHrP(102-111) peptides efficiently induced peptide-specific cytotoxic T lymphocytes from peripheral blood mononuclear cells (PBMCs) of HLA-A24(+) prostate cancer patients. Peptide-stimulated PBMCs showed cytotoxicity against prostate cancer cells in an HLA-A24-restricted manner. Experiments using antibodies and cold inhibition targets confirmed that their cytotoxicity was dependent on PTHrP peptide-specific and CD8(+) T cells. Immunoglobulin G reactive to the PTHrP(102-111) or PTHrP(110-119) peptide was frequently detected in the plasma of prostate cancer patients, suggesting that the PTHrP(102-111) peptide is able to elicit cellular and humoral immune responses in cancer patients. These results indicate that the PTHrP could be a promising target molecule for specific immunotherapy of HLA-A24(+) prostate cancer patients with metastases.

Susceptibilities of Neisseria gonorrhoeae to fluoroquinolones and other antimicrobial agents in Hyogo and Osaka, Japan.

OBJECTIVES: Decreasing susceptibility of Neisseria gonorrhoeae to fluoroquinolones has been reported in several countries. Knowledge of local N gonorrhoeae susceptibilities to various antimicrobials is important for establishing a rational treatment strategy in each region. METHODS: Isolates of N gonorrhoeae from male urethritis patients attending four urological clinics in Hyogo and Osaka prefectures in Japan were collected during 2002. The MICs for nine antimicrobials: penicillin G, tetracycline, cefixime, ceftriaxone, levofloxacin, gatifloxacin, ciprofloxacin, moxifloxacin, and spectinomycin were determined for each isolate. All isolates were also tested for beta lactamase producing profiles. RESULTS: Among the 87 isolates obtained, only one isolate was revealed to produce beta lactamase. MIC90 values for ciprofloxacin, levofloxacin, gatifloxacin, and moxifloxacin were over 8 microg/ml, over 8 microg/ml, 4 microg/ml, and 2 microg/ml, respectively. The proportion of isolates resistant to fluoroquinolones was over 60% (ciprofloxacin, 70.1%; levofloxacin, 65.5%; gatifloxacin, 70.1%). Chromosomally mediated penicillin and tetracycline resistance was identified in 12.6% and 33.3% of the isolates. MIC90 values for cefixime and ceftriaxone and were 0.5 microg/ml and 0.0063 microg/ml. All isolates were sensitive to ceftriaxone and 90.8% of them were sensitive to cefixime. MIC90 for spectinomycin was 32 microg/ml and all isolates were sensitive to it. Fluoroquinolone resistance correlated significantly with MICs for penicillin G but not tetracycline. CONCLUSION: Ceftriaxone and spectinomycin demonstrated lower MICs and so are recommended for N gonorrhoeae. Susceptibilities of N gonorrhoeae should be monitored periodically by region.

The growth inhibitory effect of p21 adenovirus on androgen-dependent and -independent human prostate cancer cells.

OBJECTIVE: To assess the potential of p21 as a gene therapy treatment for prostate cancer, by introducing p21 into both androgen-dependent (AD) and -independent (AI) human prostate cancer cell lines via a recombinant adenoviral vector, Ad5CMV-p21, carrying human p21 cDNA. MATERIALS AND METHODS: The LNCaP, DU145 and PC-3 human prostate cancer cell lines were cultured and infected with Ad5CMV-p21. Cell growth, cell-cycle progression and tumorigenicity were then assessed by thymidine incorporation into cellular DNA, and cell number, flow cytometry, and tumour growth after inoculating the cells into nude mice. RESULTS: Growth was inhibited in Ad5CMV-p21 viral-infected AD and AI prostate cancer cells. The effects were dose-dependent, regardless of the androgen status of the cell lines. Flow cytometric analysis showed that Ad5CMV-p21 arrested cell-cycle progression at G1/S with no appreciable effect on the levels of apoptotic cells. The tumorigenicity of cancer cells infected with Ad5CMV-p21 was greatly reduced in athymic mice. CONCLUSIONS: These results suggest that Ad5CMV-p21 may be a new therapeutic agent for human prostate cancer gene therapy.

Increased urinary 8-hydroxy-2'-deoxyguanosine excretion after ileal neobladder replacement.

OBJECTIVES: To examine whether orthotopic neobladder replacement using either ileum or colon segments results in increased oxidative stress, by measuring urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), one of the most commonly used markers for evaluating oxidative DNA damage. PATIENTS, SUBJECTS AND METHODS: Urinary levels of 8-OHdG and creatinine, urine analysis, nutritional status, and acid-base and electrolyte balances, were assessed in 22 patients with an ileal neobladder, 28 with a colon neobladder, 37 with an ileal conduit and 22 healthy volunteers. The results from both types of orthotopic neobladder, the ileal conduit and in the healthy controls were compared. RESULTS: The mean (sd) ratios of urinary 8-OHdG to urinary creatinine in patients with an ileal neobladder, colon neobladder, ileal conduit and in controls were 20.4 (7.8), 15.2 (4.3), 15.9 (5.1) and 15.2 (5.4) ng/mg, respectively. The urinary 8-OHdG ratio in the first group was significantly higher than in the other three groups. Among patients with a neobladder, the urinary 8-OHdG ratio was closely associated with the degree of pyuria, but not age, gender, the interval from surgery, body weight, height, serum creatinine or the degree of metabolic acidosis. CONCLUSIONS: These findings suggest that creating an ileal neobladder caused significantly greater oxidative stress than a colon neobladder, ileal conduit, or that in healthy controls. Therefore, it is recommended to conduct a careful long-term follow-up considering the possible development of malignant disease after urinary diversion, especially by an ileal neobladder.

Changes in sodium transport in epithelial cells in a rat ileal-augmented bladder model.

OBJECTIVE: To investigate changes in the morphology and sodium transport ability of intestinal epithelium diverted to the urinary tract, using an in vitro sodium-binding benzofuran isophthalate (SBFI) technique, as the effects of long-term urine exposure on the transport of electrolytes through intestine are incompletely understood. MATERIALS AND METHODS: Ileal augmentation cystoplasty was conducted in female Sprague-Dawley rats; at 3 and 12 months after surgery the serum concentration of sodium, chloride and potassium were measured. Sodium transport in the ileal epithelial cells diverted to the urinary tract was evaluated using SBFI, as the value of the 340/380 nm excitation ratio measured with fluorescence spectrophotometry. The villous height and the number of villi per ileal length were obtained from haematoxylin and eosin-stained sections. RESULTS: After 3 months the mean (sd) serum sodium concentrations in normal and augmented rats were 140.4 (2.5) and 140.7 (3.5) mmol/L, respectively; the chloride concentration in normal rats was 97.0 (2.9), and in augmented rats at 3 and 12 months it was 102.4 (2.9) and 99.0 (3.7) mmol/L, respectively. At 3 months, chloride concentrations were significantly higher in augmented than in normal rats (P < 0.05). The mean (sd) 340/380 nm ratio increased by 0.85 (0.09) in the normal ileum, and by 0.73 (0.15) and 0.49 (0.23) in the ileum of augmented rats at 3 and 12 months, respectively; the difference between normal and augmented ileum at 12 months was significant (P < 0.05). At 12 months the villous height in the augmented ileum, at 227.6 (16.0) micro m, was significantly less than in the normal ileum, at 803.4 (66.2) micro m (P < 0.05). However, the number of villi/mm ileum in normal and augmented rats at 12 months was 13.7 (1.5) and 15.0 (0.8), respectively, and not significantly different. CONCLUSION: Sodium transport decreased significantly after long-term exposure to urine; the improvement in metabolic change was probably attributable to alterations of electrolyte transport and atrophic changes of the villus.

Primary embryonal carcinoma of heart with SVC syndrome.

We present a case report of a 25-year-old man with embryonal carcinoma of right atrium and multiple lung metastases featuring SVC syndrome. We resected the cardiac tumor which occupied the right atrium and performed left upper lobectomy. No tumor mass or vestige was detected in the testes. Cis-platinum based combination chemotherapy was performed for residual lung tumors, which leads to the complete remission.

Apoptosis-related proteins in the testes of infertile men with varicocele.

OBJECTIVES: To assess immunohistochemically the expression of apoptosis-related proteins in the testes of infertile men, to determine which of these proteins were related to hypospermatogenesis, as a previous report suggested that apoptosis was suppressed in infertile men with varicocele. MATERIALS AND METHODS: The expression of Bcl-2, Bax, caspase-1 (ICE) and caspase-3 (CPP32) were examined in bilateral testicular specimens from 26 infertile men with varicocele and six normal testicular specimens, using the avidin-biotin-peroxidase complex method. Clinical variables were also assessed. RESULTS: Bax, ICE, and CPP32 were expressed in germ cells, while Bcl-2 was not. Differences in staining in left or right testes were not significant. In both testes of infertile patients with varicocele, significantly fewer germ cells stained for CPP32 than in controls (P < 0.001). For Bax and ICE, total germ cell staining was similar between these groups. Staining was less frequent in infertile patients for both CPP32 and ICE when the analysis was restricted to spermatogonia. Serum luteinizing hormone levels correlated positively with CPP32 staining (P = 0.0457). CONCLUSIONS: The reduced expression of CPP32 participates in regulating apoptosis in the testes of infertile men with varicocele.

Health-related quality of life after radical cystectomy for bladder cancer: a comparison of ileal conduit and orthotopic bladder replacement.

OBJECTIVE: To compare the health-related quality of life (HRQoL) after radical cystectomy in patients with an ileal conduit or an orthotopic neobladder. PATIENTS AND METHODS: The study included 85 men who underwent radical cystectomy for bladder cancer, comprising 48 with an orthotopic neobladder (26 with an ileal and 22 with a colon neobladder) and 37 with an ileal conduit. HRQoL was evaluated using the Short Form-36 survey containing 36 questions assessing eight aspects, including physical functioning, role-physical functioning, bodily pain, general health, vitality, social functioning, role-emotional functioning and mental health. RESULTS: The mean follow-up periods for patients with a neobladder (ileal and sigmoid) and with an ileal conduit was 45.9 (38.2 and 53.1, respectively) and 130.9 months, respectively. Scale scores were not affected by the duration of follow-up in either group. There was no significant difference in any scale scores between the neobladder and ileal conduit groups. However, general health and social functioning in both the neobladder and ileal conduit groups appeared to be significantly lower than those in the general population in the USA. Furthermore, patients with a colon neobladder had a significantly higher score for role-emotional functioning than those with an ileal neobladder, while there was no significant difference in the remaining seven scores between patients with ileal and colon neobladders. CONCLUSIONS: Six of the eight scales of HRQoL were favourable in both patients with a neobladder or an ileal conduit, and there was no significant difference between these groups. In addition, the HRQoL of patients with an orthotopic neobladder (except for role-emotional functioning) was unaffected by the segment of the intestine used for neobladder construction. Therefore, patients with both types of urinary diversion were generally satisfied with their overall health and quality of life.

Evaluation of health-related quality of life in patients treated for erectile dysfunction with viagra (sildenafil citrate) using SF-36 score.

The effect of sildenafil on health-related quality of life (HRQOL) was evaluated. Forty men (61+/-8 years old, mean +/- SD) with erectile dysfunction (ED) were studied. Sexual satisfaction as well as HRQOL according to SF-36 scales was evaluated before and after treatment with Viagra (Sildenafil citrate) at a dose of 25 or 50 mg. After treatment with Viagra, scores increased for all scales; of these, physical functioning (PF), general health (GH), and role-emotional functioning (RE) showed significant differences from baseline scores. Patients who evaluated effectiveness of the treatment as excellent showed significantly better PF, role-physical functioning (RP), and GH scale scores than those who evaluated their outcome as good or poor (p<.05 for RP and GH; p < .01 in PF). Incomparison with the scores at baseline, patients who considered outcome of Viagra to be excellent also showed significant improvements in PF, GH, and RE. Viagra treatment can improve HRQOL as well as sexual function in many patients.

Additional gene therapy with Ad5CMV-p53 enhanced the efficacy of radiotherapy in human prostate cancer cells.

PURPOSE: The aim of this study was to investigate the efficacy of combination therapy of ionizing radiation (IR) and adenoviral p53 gene therapy and to evaluate its molecular mechanisms. METHODS AND MATERIALS: Two human prostate cancer cell lines, DU145 and PC-3 cells, containing different types of p53 gene mutations, were investigated. The recombinant adenovirus vector containing the wild-type p53 gene (Ad5CMV-p53) was used for this study. Cells were irradiated (in 0, 2, 4, and 6 Gy, 300 cGy/min) and after 12 h of irradiation, the cells were infected with various doses of Ad5CMV-p53 (0-40 multiplicity of infection [MOI]). Cytotoxicity was determined by clonogenic assay. The molecular mechanisms were evaluated by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), apoptotic cell detection, and cell cycle analysis. RESULTS: The cell growth inhibition in DU145 (p53-mutated) cells by IR was strongly enhanced by additional Ad5CMV-p53 infection in a viral dose-dependent manner. In DU145 cells, IR alone induced minimal p53 mRNA expression. However, IR combined with Ad5CMV-p53 infection stimulated significant increase in p53 mRNA expression supplemented with Bax and p21 mRNA expressions. In PC-3 (p53-null), IR induced Bax and p21 mRNA expression, while the combination effects were observed in p53, Bax, and p21 mRNA expression. Apoptotic cell deaths were rarely observed after IR alone (DU145: 3%, PC-3: 5%). However, after combination therapy, the proportion of apoptotic cells greatly increased (sevenfold in DU145 cells, and twice in PC-3 cells). G1 cell cycle arrest was observed after Ad5CMV-p53 infection and the combination in both cell lines. CONCLUSION: In this study, we demonstrated that the combination of IR and Ad5CMV-p53 gene therapy resulted in remarkable synergistic effects in human prostate cancer cells. This combination therapy could be one of the optimal treatment strategies for radioresistant prostate cancer.

Synergistic chemsensitization and inhibition of tumor growth and metastasis by the antisense oligodeoxynucleotide targeting clusterin gene in a human bladder cancer model.

Clusterin expression is highly up-regulated in several normal and malignant tissues undergoing apoptosis. Although recent studies have demonstrated a protective role of clusterin expression against various kinds of apoptotic stimuli, the functional role of clusterin in the acquisition of a therapy-resistant phenotype in bladder cancer remains unknown. The objectives of this study were to determine whether antisense (AS) oligodeoxynucleotide (ODN) targeting the clusterin gene enhances apoptosis induced by cisplatin and to evaluate the usefulness of combined treatment with AS clusterin ODN and cisplatin in the inhibition of KoTCC-1 tumor growth and metastasis in a human bladder cancer KoTCC-1 model. We initially revealed the dose-dependent and sequence-specific inhibition of clusterin expression by AS clusterin ODN treatment in KoTCC-1 cells at both mRNA and protein levels. Clusterin mRNA was increased in a dose-dependent manner by cisplatin treatment at concentrations < or =10 mg/ml, and clusterin mRNA up-regulation induced by 10 mg/ml cisplatin peaked by 48-h post-treatment and began decreasing by 72-h post-treatment. Although there was no significant effect on growth of KoTCC-1 cells, AS clusterin ODN treatment significantly enhanced cisplatin chemosensitivity of KoTCC-1 cells in a dose-dependent manner, reducing the IC(50) by >50%. Characteristic apoptotic DNA ladder formation and cleavage of poly(ADP-ribose) polymerase protein were detected after combined treatment with AS clusterin ODN and cisplatin but not either agent alone. In vivo systemic administration of AS clusterin and cisplatin significantly decreased the s.c. KoTCC-1 tumor volume compared with mismatch control ODN plus cisplatin. Furthermore, after the orthotopic implantation of KoTCC-1 cells, combined treatment with AS clusterin and cisplatin significantly inhibited the growth of primary KoTCC-1 tumors, as well as the incidence of lymph node metastasis. Collectively, these findings demonstrated that clusterin helps confer a chemoresistant phenotype through inhibition of apoptosis and that combined AS clusterin ODN may be useful in enhancing the effects of cytotoxic chemotherapy in patients with bladder cancer.

Gene therapy for bladder cancer using adenoviral vector.

BACKGROUND AND PURPOSE: Bladder cancer is common. Current treatment for patients with superficial bladder cancer involves transurethral resection followed by adjuvant bacillus Calmette-Guérin (BCG) administration. Adjuvant BCG has been reported to be effective in 38% to 68% of patients; however, more than 30% of patients do not respond. Because p53 mutations are common among superficial bladder cancers, we tested the feasibility of using p53 as a gene therapy agent for targeting superficial tumors, which are easily accessible using an intravesical approach. MATERIALS AND METHODS: Wild-type p53 was transduced into various human and murine bladder cancer cell lines (HTB9, KU-1, and MBT-2) using a recombinant adenoviral vector (Ad5CMV-p53) in vitro. Also, subcutaneous tumors were established and then treated with intratumoral injection of Ad5CMV-p53 or control viruses. RESULTS: In vitro assays revealed significant growth suppression of target cells by Ad5CMV-p53 in comparison with those receiving the control Ad5-CMV-PA vector or untreated control cells. In vivo studies using subcutaneous bladder tumor models established in syngeneic mice demonstrated that the rate of tumor growth and volume was reduced to a greater extent by 14 days of intratumoral injection of Ad5CMV-p53 rather than Ad5CMV-PA. Furthermore, the survival of host animals bearing tumors that were infected with Ad5CMV-p53 was significantly longer than that of the control group treated with Ad5CMV-PA (P < 0.01). CONCLUSION: Our data suggest that Ad5CMV-p53 is effective in suppressing bladder cancer growth and improving host survival.

Acquisition of resistance to Fas-mediated apoptosis by overexpression of clusterin in human renal-cell carcinoma cells.

Recent studies have shown the antiapoptotic activity of clusterin against a wide variety of stimuli; however, the functional role of clusterin in Fas-mediated apoptosis has not been well characterized. We transfected the clusterin cDNA into human renal-cell carcinoma (RCC) ACHN cells that scarcely express clusterin protein in order to examine whether overexpression of clusterin inhibits the Fas-mediated signal pathway for apoptotic cell death. No significant difference was observed in the in vitro cell growth rates between the clusterin-transfected cell line (ACHN/CL) and the vector-only-transfected control cell line (ACHN/C), whereas the colony-forming efficiency in soft agar of ACHN/CL was significantly higher than that of ACHAN/C. The anti-Fas monoclonal antibody CH11 induced apoptosis in ACHAN/C cells in a dose-dependent manner; however, the growth-inhibitory effect of CH11 on ACHN/CL cells was markedly suppressed, with corresponding increases in p53 expression and decrease in the fraction of cells in the sub-G(1) phase of the cell cycle. Furthermore, the cytotoxic effect of CH11 on ACHN/CL cells was augmented by treatment with interferon-gamma, but a corresponding effect on ACHN/C cells was not observed. These findings suggest that overexpression of clusterin may contribute to a phenotype resistant to Fas-mediated apoptosis, and that if interferon-gamma treatment is added according to the clusterin expression level, Fas-mediated therapy could be a novel approach to RCC.

Serum inhibin pro alphaC in infertile men.

Specific assays have been developed for bioactive inhibin dimers, inhibin A and B, and inhibin alpha-subunit precursor pro alphaC. To better understand the role of serum inhibin pro alphaC in infertile men, the authors measured these forms of inhibin in sera from 39 infertile men and analyzed inhibin relationships with serum gonadotropins, testosterone, and estradiol. All subjects had oligozoospermia. Inhibin A levels were undetectable in all subjects. Inhibin B concentrations were 117 +/- 59 pg/mL. Inhibit B concentrations correlated negatively with serum FSH (r = .584, p < .0001) and positively with sperm count (p < .01) and bilateral testicular volume (r = .607, p < .0001). The concentration of pro alphaC was 556 +/- 236 pg/mL (normal range, 446 +/- 28). Inhibin pro alphaC showed no correlation with serum FSH, LH, testosterone, sperm concentration, and bilateral testicular volume. In addition, inhibin pro alphaC was not correlated with inhibin B. Pro alphaC is unlikely to be a useful marker for spermatogenesis in infertile men compared with inhibin B.

Inhibition of steroidogenesis in Leydig cells by exogenous nitric oxide occurs independently of steroidogenic acute regulatory protein (star) mRNA.

Nitric oxide (NO) plays multiple roles in the reproductive system. The authors studied the effect of NO on LH-stimulated steroidogenesis in primary cultures of rat Leydig cells, particularly seeking a link between inhibition of steroidogenesis and changes in expression of steroidogenic acute regulatory protein (StAR). Sodium nitroprusside (SNP), an NO generator, did not alter basal testosterone, but dose-dependently reduced testosterone production in the Leydig cells stimulated by LH (100 ng/mL) at 3 h after addition of SNP. Induction of StAR mRNA transcripts could be detected as early as 1 h after the addition of LH, but no effect was detected of SNP on LH induction of StAR mRNA. StAR, then, is not affected in the inhibition by NO of LH-stimulated steroidogenesis in Leydig cells.

Significance of prostate-specific antigen--alpha(1)-antichymotrypsin complex for diagnosis and staging of prostate cancer.

OBJECTIVE: To evaluate the clinical significance of measuring the prostate-specific antigen-alpha(1)-antichymotrypsin (PSA-ACT) for differentiating prostate cancer from benign prostate hypertrophy (BPH) and for the staging of prostate cancer. METHODS: Before treatment, total PSA (tPSA) and PSA-ACT were measured in 120 patients with prostate cancer and in 150 patients with BPH using immunofluorometric techniques with different monoclonal antibodies against PSA and ACT. Furthermore, the tPSA and PSA-ACT densities of the whole prostate (PSAD and ACTD, respectively) were calculated. RESULTS: tPSA, PSAD, PSA-ACT and ACTD levels in patients with prostate cancer paralleled the clinical stage and were significantly higher than those in patients with BPH. Furthermore, these four values were significantly higher in patients with pathologically extraprostatic disease than those with organ-confined disease. Receiver operating characteristics analysis among patients with PSA values of 4.1-10 ng/ml revealed that the areas under the curve for tPSA and ACTD were similar to those for PSA-ACT and ACTD, respectively and that no significant differences in the differentiation between prostate cancer and BPH were observed among these parameters. CONCLUSIONS: Measurement of PSA-ACT provides useful information for the clinical staging of prostate cancer and differential diagnosis between prostate cancer and BPH; however, compared with tPSA, PSA-ACT may not be significantly superior in the diagnosis and staging of prostate cancer.

Fluorescence in situ hybridization analysis of sex-chromosome mosaicism in azoospermic men.

A retrospective study was carried out to determine the prevalence of sex-chromosome mosaicism among azoospermic men and to evaluate the feasibility of using fluorescence in situ hybridization (FISH) technique to assess mosaicism and the origin of marker chromosomes. Nine hundred eighty patients with azoospermia who were referred to a male infertility clinic at a university hospital were karyotyped by G-banding using peripheral blood lymphocyte (PBL) metaphase spreads. When sex chromosome mosaic karyotype was detected, FISH analyses using sex chromosome-specific probes were performed. Seventeen of 980 patients showed evidence of sex chromosomal mosaic karyotype or mosaicism with marker chromosomes by G-banding studies of PBLs. Ten patients showed mosaicism in the number of sex chromosomes and 7 showed mosaicism with marker chromosomes. All 17 patients agreed to undergo FISH analysis. FISH confirmed mosaicism in 88.2% of these patients (15 of 17). Low-frequency mosaicism showing a frequency of less than 10% by G-banding was proved to be nonmosaicism by FISH. Marker chromosomes detected in 7 patients were proved to be derived from the Y chromosome by FISH analyses. From these data the prevalence of sex chromosome mosaicism confirmed by FISH analysis is 1.5% (15 of 980 patients). FISH analysis should be applied when mosaicism shows a frequency of less than 10% by the G-banding technique. Also, FISH analysis is indicated when a marker chromosome is detected by G-banding.