A phase I/II study of cancer peptide vaccine S-288310 in patients with advanced urothelial carcinoma of the bladder.

Background: S-288310, a cancer peptide vaccine composed of two HLA-A*24:02-restricted peptides derived from two oncoantigens, DEP domain-containing 1 (DEPDC1) and M-phase phosphoprotein 1 (MPHOSPH1), was investigated in urothelial carcinoma (UC) of the bladder. Patients and methods: Thirty eight HLA-A*24:02-positive patients with progressive UC were enrolled in this study. In the phase I part of the study, three patients each were treated with S-288310 at 1 mg or 2 mg/peptide subcutaneously once a week to evaluate safety and tolerability. In the phase II, 32 patients were randomized to receive either 1 mg or 2 mg to evaluate the difference in cytotoxic T lymphocytes (CTL) induction and safety. Results: S-288310 was safe and well tolerated in the phase I. Of 27 patients evaluable for immune responses in the phase II, there was no difference in CTL induction rate between the 1 mg (100%) and 2 mg (80.0%) groups. Of 32 patients receiving S-288310 in the phase II, the most frequent drug-related AE was the injection site reaction that was observed in 29 patients (90.6%), but none of the patients discontinued administration due to these reactions and no dose relationship in the frequency and severity was observed. The objective response rate of the 32 patients was 6.3% and the disease control rate was 56.3%. The median overall survival (OS) rates for patients vaccinated with S-288310 after one regimen of chemotherapy, 2 regimens, or 3 or more were 14.4, 9.1 and 3.7 months, respectively, and 32.2% of patients post first-line treatment were alive at 2 years. OS of patients who showed CTL induction to both peptides was longer than that of those with CTL induction to no or one peptide. Conclusion: S-288310 was well-tolerated and effectively induced peptide-specific CTLs, which were correlated with longer survival for patients with UC of the bladder. Trial registration ID: JapicCTI-090980.

Total skin electron beam therapy using an inclinable couch on motorized table and a compensating filter.

Total skin electron beam is a specialized technique that involves irradiating the entire skin from the skin surface to only a few millimetres in depth. In the Stanford technique, the patient is in a standing position and six different directional positions are used during treatment. Our technique uses large electron beams in six directions with an inclinable couch on motorized table and a compensating filter was also used to spread the electron beam and move its intensity peak. Dose uniformity measurements were performed using Gafchromic films which indicated that the surface dose was 2.04 ± 0.05 Gy. This technique can ensure the dose reproducibility because the patient is fixed in place using an inclinable couch on a motorized table.

An isocenter detection and verification device for use in proton therapy.

We developed a new device for isocenter detection and verification for use in proton therapy. This device can be used to confirm the isocenters of a room laser, a digital radiography system, and a rotational gantry. Agreement between the isocenters of a room laser and a digital radiography system was examined visually. We used the star-shots method for detecting and verifying the rotational gantry isocenter. The gafchromic film could be irradiated with this method using a 2 mm slitted 200 MeV proton beam. The isocenters of a room laser and digital radiography system were in good agreement. The size distribution of the isocenter by the star-shots method was <2 mm, which is the value recommended by the American Association of Physicists in Medicine Task Group 142. This new device can be used to evaluate comprehensive isocenter detection and verification for related equipment. Subsequent checking using this new device will decrease the uncertainty in measuring an isocenter. When using this device, the working time was significantly reduced to about 30 min, including preparation. These measurements should be useful for high accuracy daily treatments.

Regulation of gene expression in prostate cancer cells with an artificially constructed promoter responsive to radiation.

A promoter library was developed that is composed of DNA fragments constructed by randomly elongating the cis-acting elements of transcription factors presumably activated in prostate cancer by radiation, and linking to the TATA-box sequence. One promoter with the strongest reactivity to X-ray in the LNCap cells of the library was chosen and improved by the introduction of random mutations. The resultant promoter was designated clone 880-8, showing the highest dose-dependent activity enhancement with X-ray irradiation (X-irradiation). A recombinant retrovirus expressing the luciferase gene under the control of clone 880-8 was infected into LNCap cells that showed 9.12±0.36-fold enhancement of luciferase activity 12 h after X-irradiation at 10 Gy. When the infected cells were inoculated onto nude mice, enhancement of luciferase expression was 4.27±1.36-fold 12 h after X-irradiation at 10 Gy. When LNCap was infected with another recombinant carrying the fcy::fur gene downstream from clone 880-8, fcy::fur expression was enhanced by X-irradiation. It was also shown to increase the dose-dependent cell killing ratio with 5-FC as compared with a counterpart without X-irradiation. These results suggest that the method used in this study is effective to construct a promoter responsive to stimulation. Such promoters can be used for stimulation-controlled gene therapies.

Activities and distribution of methanogenic and methane-oxidizing microbes in marine sediments from the Cascadia Margin.

We investigated methane production and oxidation and the depth distribution and phylogenetic affiliation of a functional gene for methanogenesis, methyl coenzyme M reductase subunit A (mcrA), at two sites of the Integrated Ocean Drilling Program Expedition 311. These sites, U1327 and U1329, are respectively inside and outside the area of gas hydrate distribution on the Cascadia Margin. Radiotracer experiments using (14)C-labelled substrates indicated high potential methane production rates in hydrate-bearing sediments [128-223 m below seafloor (mbsf)] at U1327 and in sediments between 70 and 140 mbsf at U1329. Tracer-free experiments indicated high cumulative methane production in sediments within and below the gas hydrate layer at U1327 and in sediments below 70 mbsf at U1329. Stable tracer experiments using (13)C-labelled methane showed high potential methane oxidation rates in near-surface sediments and in sediments deeper than 100 mbsf at both sites. Results of polymerase chain reaction amplification of mcrA in DNA were mostly consistent with methane production: relatively strong mcrA amplification was detected in the gas hydrate-bearing sediments at U1327, whereas at U1329, it was mainly detected in sediments from around the bottom-simulating reflector (126 mbsf). Phylogenetic analysis of mcrA separated it into four phylotype clusters: two clusters of methanogens, Methanosarcinales and Methanobacteriales, and two clusters of anaerobic methanotrophic archaea, ANME-I and ANME-II groups, supporting the activity measurement results. These results reveal that in situ methanogenesis in deep sediments probably contributes to gas hydrate formation and are inconsistent with the geochemical model that microbial methane currently being generated in shallow sediments migrates downward and contributes to the hydrate formation. At Site U1327, gas hydrates occurred in turbidite sediments, which were absent at Site U1329, suggesting that a geological setting suitable for a gas hydrate reservoir is more important for the accumulation of gas hydrate than microbiological properties.

Postoperative changes of sperm chromatin heterogeneity, using acridine orange staining, in varicocele patients.

The aim of this study was to evaluate postoperative changes in sperm chromatin heterogeneity in varicocele patients. In 15 infertile patients with varicocele, sperm parameters including concentration, motility, and morphology were evaluated before and after surgical correction of varicocele. Sperm motion analysis using computer-assisted semen analyzer (CASA) was also performed. To analyze the sperm nuclear proteins, the acridine orange staining method was used. On semen analysis, sperm concentration and motility significantly increased after surgery (p = 0.002, p = 0.003, respectively), although sperm morphology was unaltered postoperatively. CASA parameters, including velocity, linearity, amplitude of lateral head displacement and beat cross frequency were unaltered postoperatively. On the other hand, acridine orange staining significantly increased postoperatively (p = 0.002). Varicocele influences the sperm chromatin condition, as well as sperm concentration and motility.

Treatment with prednisolone of hormone-refractory prostate cancer.

Fifteen patients 60 to 80 years old (a mean of 72 years) with hormone-refractory prostate cancer were treated with low dose prednisolone. All patients had previously undergone hormone therapy. Prostate specific antigen (PSA) values decreased in 11 cases (73%), of which 4 had PSA decreases of 50% or greater. Serum levels of DHEAS significantly decreased at 4 and 8 weeks after treatment (both intervals were p < 0.05 vs pretreatment). Of 8 patients with bone metastasis evaluation, 2 (25%) showed improvement of the lesion. In 5 patients (33%), relief of pain was observed one month after starting prednisolone. The one-year survival rate was 58%. The side effects were mild and manageable in an outpatient clinic.

Usefulness of sperm quality analyzer-V (SQA-V) for the assessment of sperm quality in infertile men.

The aim of this study was to evaluate the correlation of new of Sperm Quality Analyzer (SQA-V) with the computer-assisted sperm analysis (CASA) and manual semen analysis estimates. One hundred five fresh semen samples were analyzed using SQA-V and CASA and manual semen analysis. Significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and normal morphology (p < 0.0001) were observed between SQA-V variables and manual semen analysis estimates. There also were significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and sperm velocity (p = 0.0235) between SQA-V variables and CASA estimates. Meanwhile, it did not correlate with amplitude of lateral head displacement, beat cross frequency, lineality assessed by CASA. The value of the sperm concentration and the sperm motility measured by SQA-V showed high correlations with the value of those measured by CASA and manual semen analysis. In addition, velocity and sperm morphology may also be evaluated to some extent using SQA-V.

Assessment of sperm quality analyzer II B: comparison with manual semen analysis and CASA.

Two hundred and seven patients with male infertility were investigated. Total sperm concentration and percent progressive motility by SQA IIB showed high correlations with those of conventional manual method. Percent of normal morphology showed a significant correlation among these techniques. The sperm motility index (SMI) and total functional sperm concentration (TFSC) demonstrated high correlations with any variables of manual analysis. Only velocity and amplitude of lateral head displacement showed significant correlations with the variables obtained by SQA IIB, especially with SMI and TFSC. It was suggested that SQA IIB could be a useful instrument in the clinical practice of infertility as a screening test for semen quality.

Effect of Bu-zhong-yi-qi-tang on seminal plasma cytokine levels in patients with idiopathic male infertility.

Twenty-two idiopathic infertile patients were treated with Oriental herbal medicine, Bu-zhong-yi-qi-tang (Hochu-ekki-to). Seminal plasma sFas level elevated significantly after herb administration. There was an inverse correlation between seminal plasma IL-6 level and sperm concentration before treatment. After the administration of herb seminal plasma, sFas levels significantly correlated with sperm concentration. Seminal plasma IL-8 level did not change. Certain kinds of cytokine in the seminal plasma might play some role in improving semen quality with treatment.

Transurethral treatment of ejaculatory duct obstruction in infertile men.

Ten cases of ejaculatory duct obstruction with midline cyst, complaining of male infertility, were treated with transurethral incision. They had azoospermia or oligozoospermia, and physical examination did not show any abnormal findings. Serum levels of testosterone, LH, and FSH were within normal limits. The diagnosis of ejaculatory duct obstruction with midline cyst was made by transurethral ultrasonography and vasography. Thereafter, patients underwent transurethral incision of the verumontanum with a cold knife. Semen volume increased in all patients, and sperm concentration and/or motility improved in 7 patients (70%). Pregnancy was achieved by 3 couples (30%). Incision of the ejaculatory duct via an endoscopic technique could improve seminal findings and subsequent fertility.

Transurethral incision for hematospermia caused by ejaculatory duct obstruction.

Hematospermia is supposed to be derived from pathological conditions in the seminal vesicle, prostate, testis, epididymis, or urethra. A recent advance in diagnostic procedures has demonstrated the seminal vesicle, the prostate, and midline cyst as potential sources of hematospermia. The authors describe a case of hematospermia caused by ejaculatory duct obstruction, in which a transurethral technique was successful. A 51-year-old male was referred to the authors' clinic with a chief complaint of hematospermia. Transurethral ultrasonography showed a cystic lesion surrounded with hyperechoic area in the middle of the prostate. Vasography demonstrated the distal dilation of the ejaculatory duct. Magnetic resonance imaging demonstrated a high signal intensity area in the middle of the prostate. Urethrocystoscopy showed an enlarged cystic lesion with an orifice at the prostatic urethra, which was incised endoscopically. There was no complication observed postoperatively. Seven months after the technique, hematospermia resolved completely. A midline cyst should be considered a cause of hematospermia and the incidence of such cysts may be higher than that previously recognized. The transurethral technique is expected to be a successful treatment approach.

Soluble Fas and interleukin-6 and interleukin-8 levels in seminal plasma of infertile men.

The seminal plasma levels of soluble Fas (sFas) and interleukins IL-6 and IL-8 were measured and their relationship with semen qualities was examined. The seminal plasma sFas level in fertile males was significantly higher than that in the infertile group. On the other hand, seminal plasma level of IL-6 was significantly lower in fertile males than in the infertile group. In the infertile group, patients with oligozoospermia had a lower seminal plasma sFas and a higher IL-6 level than those with normal sperm concentration. There was an inverse correlation between IL-6 and sperm concentration in infertile patients. Seminal plasma IL-8 was not correlated with sperm parameters. It would appear that certain kinds of cytokine in the seminal plasma might play an important role in improving semen quality.

Persistent Müllerian duct syndrome: a surgical approach.

Laparoscopy revealed a left inguinal testis and a right abdominal testis. Surgery revealed uterus-like structures. The bilateral testes showed primitive testis without ovarian tissue. Physical examination showed a normal and an empty scrotum with a nonpalpable gonad. Chromosome analysis revealed 46,XY. Pathological findings demonstrated the immature testis and the immature uterus.

Evaluation of acrosome reactivity using the Acrobeads test in varicocele patients: findings before and after treatment.

The Acrobeads test was performed on semen samples from 43 patients with varicocele before and after varicocelectomy. Sperm motility significantly increased after surgery (p <.05), while sperm concentration and motile sperm concentration did not alter postoperatively. Sperm motion analysis using CellSoft 3000 did not demonstrate a significant change after treatment. Acrobeads score significantly increased after the procedure (p<.005). Pregnancy was achieved in 10 patients (28%). Acrobeads score in cases that achieved pregnancy was increased postoperatively (p<.005). The percentage of patients with a postoperative increase in Acrobeads score in the group that achieved pregnancy was significantly higher than that observed in the unsuccessful group (p <.05). Sperm parameters other than the Acrobeads score did not show a significant difference between the successful and unsuccessful patients. The Acrobeads test assessed postoperatively can be useful in precisely evaluating fertility potential after varicocele repair.

Metastatic prostate cancer with normal level of serum prostate-specific antigen.

The clinical and pathological features of metastatic prostate cancer with normal level of serum prostate-specific antigen (PSA) were investigated. Four patients with metastatic prostate cancer had serum PSA within the normal range at the diagnosis. All tumors were poorly-differentiated adenocarcinoma. Endocrine therapy was performed as the initial therapy in all patients. Despite subsequently treatment, all cases died of prostate cancer at 2, 8, 9 and 38 months. During disease progression, 3 of 4 patients had elevated serum markers such as carcinoembryonic antigen (CEA), CA19-9, CA15-3, CA125, neuron-specific enolase and pro-gastrin releasing peptide. Immunohistochemical examination of the initial biopsy specimens revealed that 4 and 3 cases were positive for CEA and chromogranin A, respectively. In advanced prostate cancer patients with low PSA level, those markers may aid in the follow up of disease.

DNA microarray analysis of genes involved in the process of differentiation in mouse Leydig cell line TTE1.

A Leydig cell line, TTE1, was established from the temperature-sensitive simian virus 40 large T-antigen transgenic mice. The cells showed temperature-sensitive growth characteristics and a differentiated phenotype at a nonpermissive temperature. To identify differentially expressed genes in the process of Leydig cell differentiation, the authors carried out microarray analysis of TTE1 cells cultured at permissive and nonpermissive temperatures. The resulting fluorescence-labeled cDNAs synthesized from mRNAs were hybridized with Clontech's Atlas glass mouse 1.0 microarrays. Of the 1081 genes analyzed, the levels of 31 genes were changed, with 24 genes showing increased levels of expression and the remaining 7 genes showing decreased levels. Tie2 was the most changed transcript, with a 13.5-fold upregulation under the differentiated condition. The authors believe this to be the first report of broadscale gene expression in Leydig cell differentiation using the microarray technology. The ability to analyze broadscale gene expression in this manner provides a powerful tool for investigating the molecular mechanisms of Leydig cell functions.

Establishment of Leydig cell line, TTE1, from transgenic mice harboring temperature-sensitive simian virus 40 large T-antigen gene.

A Leydig cell line, TTE1, has been established from transgenic mice harboring a temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene. The cells grew at a permissive temperature (33 degrees C), but growth was markedly prevented at a nonpermissive temperature (39 degrees C). T-antigen was expressed in the nuclei at 33 degrees C but disappeared at 39 degrees C, indicating that the cells show a temperature-sensitive growth phenotype reflected by the tsSV40 large T-antigen. TTE1 cells did not show any colony-forming activity in soft agar and form tumors in subcutaneous tissue in nude mice, indicating that the cells were not transformed. Alkaline phosphatase and 3beta-hydroxysteroid dehydrogenase (HSD) activities or expression of cytokeratin and vimentin were observed. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that TTE1 cells expressed mRNAs encoding 17beta-HSD types 1 and 3, and inhibin-alpha. The cells with unique characteristics, therefore, should serve useful model study the function of Leydig cell.

Tubularized incised plate urethroplasty for distal hypospadia, using overlapping dorsal subcutaneous flaps.

This paper presents a newly developed simple procedure using overlapping vascularized dorsal dartos subcutaneous flaps to cover the neourethra after hypospadias correction. A 3-year old boy with distal hypospadias underwent the tubularized incised plate urethroplasty using this method. Postoperative complications, which include urethrocutaneous fistula, were not observed, but the glans did exhibit a vertical, slit-like meatus. Covering the neourethra with overlapping dorsal dartos flaps is expected to prevent the development of urethrocutaneous fistula in patients with distal hypospadias.

Transitional cell carcinoma of the renal pelvis in a long-term hemodialysis patient with autosomal dominant polycystic kidney.

We present a case of renal pelvic transitional cell carcinoma (TCC) accompanied by autosomal dominant polycystic kidney disease (ADPKD). An 81-year-old woman on long-term hemodialysis for ADPKD visited the Department of Urology at Kouseiren Takaoko Hospital, complaining of asymptomatic macroscopic hematuria. Retrograde pyelogram showed an irregular filling defect in the right renal pelvis, which suggested the right renal pelvic tumor. We performed right nephrectomy and transurethral removal of the right ureter. Histological diagnosis demonstrated renal pelvic TCC.