Toshiya Akao

Hypermethylation at 9q32-33 tumour suppressor region is age-related in normal urothelium and an early and frequent alteration in bladder cancer

Transcriptional silencing by CpG island hypermethylation of gene regulatory regions is one mechanism for inactivation of tumour suppressor genes. Chromosome 9q deletion is frequently found in transitional cell carcinoma (TCC) of the bladder and upper urinary tract and one of the putative tumour suppressor loci has been mapped to 9q32-33. A gene designated as DBCCR1 was identified in the candidate region and its mRNA expression is thought to be suppressed by hypermethylation. To understand the role of hypermethylation in TCC, we evaluated the methylation status of 20 CpG sites of the DBCCR1 5′-CpG island region in a total of 69 tumours from 45 patients, 21 normal urothelial specimens, and six bladder cancer cell lines. Aberrant hypermethylation levels were found in 36 (52%) of 69 tumours without any association with tumour grade or stage. Methylation was weakly detected in the normal urothelium in association with ageing. Although recurrent tumours tended to have higher methylation levels than the initial tumours, the methylation pattern was mostly maintained between multifocal TCCs in individual patients. The results suggest that hypermethylation of the DBCCR1 region is one of the earliest alterations in the development of TCCs and there may be an age-related hypermethylation-based field defect in normal urothelium. Methylator or methylation-resistant phenotype seems to be maintained during multifocal development or recurrence of most TCCs.

DISTINCT MICROSATELLITE ALTERATIONS IN UPPER URINARY TRACT TUMORS AND SUBSEQUENT BLADDER TUMORS

PURPOSE Although synchronous and/or metachronous tumor development is common in urothelial cancer, genetic and biological differences in upper urinary tract and bladder tumors are unclear. We compared the genetic alteration pattern in multifocal disease in patients with upper urinary tract and subsequent bladder tumors, and those with recurrent bladder tumor. MATERIALS AND METHODS Using 21 microsatellite markers on the 8 chromosomal arms 2q, 4p, 4q, 8p, 9p, 9q, 11p and 17p we analyzed 34 tumors from 15 patients with upper urinary tract and subsequent bladder disease, and 70 tumors from 22 with recurrent bladder disease. RESULTS Judging from the patterns of genetic alterations multifocal tumors were considered to have derived from an identical progenitor cell in 7 of 13 evaluable patients (54%) with upper urinary tract and subsequent bladder tumors, and 16 of 19 (84%) who were evaluable with recurrent bladder tumor. These data confirm the view that seeding or intraepithelial spread is a major mechanism for the multifocal development of urothelial cancer in general. However, a discordant microsatellite alteration pattern in multifocal tumors was observed in 6 of 7 patients (86%) with upper urinary tract and subsequent bladder lesions but in 2 of 16 (13%) with recurrent bladder cancer (p <0.005). CONCLUSIONS Our results imply that upper urinary tract neoplasms may be genetically more unstable than bladder neoplasms. The implantation of tumor cells from upper urinary tract to bladder may involve additional and diverse genetic alterations. Furthermore, a considerable number of multifocal upper urinary tract and subsequent bladder lesions may arise independently via field cancerization mechanism. Our study indicates that the factors contributing to multifocal development are different in the 2 groups.

Early and large-dose intravesical instillation of epirubicin to prevent superficial bladder carcinoma recurrence after transurethral resection

To prospectively compare the prevention of tumour recurrence by four intravesical adjuvant administration protocols, and thus elucidate the efficacy of early and high total dose instillations of epirubicin to prevent superficial bladder tumour recurrence after transurethral resection of bladder tumour (TURBT).

Chrono and clinical pharmacokinetic study of tacrolimus in continuous intravenous administration

Abstract Background: The circadian variation of clinical pharmacokinetics of tacrolimus in kidney transplant recipients receiving continuous intravenous administration has not been clarified. The aim of this study was to evaluate the circadian variation of this drug in continuous intravenous administration, with regard to the dosing scheme for conversion from intravenous to oral therapy.

A High Prevalence of Functional Inactivation by Methylation Modification of p16INK4A/CDKN2/MTS1 Gene in Primary Urothelial Cancers

We analyzed the genetic and epigenetic alterations p16INK4A/CDKN2/MTSl gene (MTS1 gene) in 38 primary urothelial cancers. Genetic alterations of the MTS1 gene consisted of one base substitution mutation in exon 2(2.6%)and 6 homozygous deletions (16.2%). Hypermethylation of the 5’CpG island in exon 1 of theMTS1 genewas observed in 12 tumors (37.5%). Consequently, 19 of 38 tumors (50%) showed genetic alterations or epigenetic hypermethylation of the MTS1 gene. Retention of hypermethylated MTS1 gene(s) in 36% of the tumors showing loss of heterozygosity at the critical region indicates that the methylation modification could be an initial event followed by genomic rearrangements associated with total loss of MTS1 gene function. Immunohistochemical analysis of MTS1 expression revealed that all the tumors with genetic alterations of the MTS1 gene and 9 of 12 highly methylated tumors displayed an absence of MTS1 nuclear antigen. Genetic and epigenetic changes of theMTS1 gene were not correlated with the grade and stage of tumors, indicating that these alterations are early events in nrothelial carcinogenesis, in which functional inactivation by hypermethylation is a predominant mechanism.

BCL10 is not a major target for frequent loss of 1p in testicular germ cell tumors

The deletion of chromosome 1p is one of the frequent genetic alterations found in testicular germ cell tumors (GCTs), suggesting the presence of a tumor suppressor gene. BCL10, which was identified as a gene altered in mucosa-associated lymphoid tissue lymphoma, has been mapped at 1p22. The gene has been reported to be mutated in a variety of human cancers. In this study, we investigated the allelic deletions on 1p and the mutation of BCL10 in 51 GCTs comprising 30 seminomas and 21 non-seminomatous germ cell tumors. Loss of heterozygosity (LOH) on 1p was tested using three microsatellite markers. The search for BCL10 mutations in each of the three exons was screened by a single-stranded conformation polymorphism (SSCP) analysis and samples with abnormal bandshifts were directly sequenced. LOH at at least one locus tested was found in 42% (21/49) of the tumors (43% of seminomas and 38% of NSGCTs). SSCP and direct sequence analyses revealed that there were single nucleotide polymorphisms at codon 5, 8, 162, and intron 1. However, there were no somatic mutations of BCL10 in the 51 tumors. In support of the previous studies, our results demonstrated that LOH on 1p is frequent in both seminomas and NSGCTs, indicating that there is an important tumor suppressor on 1p in GCT. However, the results indicate that BCL10 is not a candidate target gene of the 1p deletion.

Transitional cell carcinoma in an ectopic ureter

We experienced an 82‐year‐old man with transitional cell carcinoma in an ectopic ureter draining into the prostatic urethra. Carcinoma arising from an ectopic ureter is very rare and a differential diagnosis is difficult. To our knowledge, our case is the third male case reported in the literature.

Semi-quantitative analysis of telomerase activity of exfoliated cells in urine of patients with urothelial cancers:: Causative factors affecting sensitivity and specificity

We previously reported that detection of telomerase activity in urinary exfoliated cells obtained from urothelial cancer patients by telomeric repeat amplification protocol (TRAP) assay is a more sensitive method of diagnosis than conventional urine cytologic examination, particularly in patients with grade 1 tumors. To establish this method as a noninvasive screening test for the diagnosis of urothelial cancers, we performed the semi-quantitative analysis of telomerase activity using Telomerase PCR ELISA™. Spontaneous voided urine samples were obtained from 65 urothelial and 58 non-urothelial cancer patients. When the mean + 2 standard deviation of telomerase activity in urine sediments of non-urothelial cancer patients was arbitrarily determined as a cut-off, the sensitivity of TRAP enzyme-linked immunosorbent assay (ELISA) and the conventional cytology were 57% and 35%, respectively. Detection rate was significantly higher in semi-quantitative TRAP assay than in conventional cytologic examination in grade 1 cancer patients (52% vs. 5%, p = 0.00195). False positives were detected in 5% of non-urothelial cancer patients without pyuria and in 11% of non-urothelial cancer patients with pyuria (p = 0.395). Telomerase activity was enhanced in some cases after phenol extraction or extracting epithelial cells by using Dynabeads of macroscopic hematuria and pyuria, indicating that hematuria and pyuria might contribute to false negatives. In conclusion, the TRAP-ELISA method is superior to the standard TRAP assay in quantitativeness and simplicity of the experimental procedure. Detection of telomerase activity in urine sediments is particularly useful for the diagnosis of low-grade tumors. However, telomerase activity in patients with high grade tumors often might be underestimated due to the excessive amount of exfoliated epithelia with necrotic tissues, hematuria, and pyuria.

Association of vitamin D receptor gene polymorphism with protection against prostate cancer and benign prostate hyperplasia

Association of vitamin D receptor gene polymorphism with protection against prostate cancer and benign prostate hyperplasia

Laparoscopic intracorporeal ileal conduit after laparoscopic radical cystectomy: A modified technique to facilitate ureteroenteric anastomosis

DOI: 10.1111/iju.13782 ICUD has the potential benefits of a smaller incision, reduced pain, decreased bowel exposure, reduced risk of fluid imbalance and early postoperative recovery. Indeed, ICUD is gaining popularity during RARC, but purely laparoscopic ICUD has seldom been reported because of the technical difficulty, particularly precise intracorporeal suturing. We herein describe our modified technique to facilitate ureteroenteric anastomosis during pure laparoscopic intracorporeal IC after LRC and evaluate perioperative outcomes. From 2007 to 2017, 66 patients underwent LRC with pelvic lymphadenectomy with IC diversion at three institutions (35 intracorporeal IC, 31 extracorporeal). Total intracorporeal IC began in May 2014. LRC started using a five-port, fan shaped, transperitoneal approach. After lymphadenectomy and radical cystectomy, the left ureter was delivered under the sigmoid mesocolon to the right side. A 15-cm ileal segment of the bowel was harvested as previously described during RARC. The schema of ureteroenteric anastomosis and intraoperative findings are shown in Figure 1 and Video S1. After inserting a guidewire from a 3-mm port placed just superior to the pubic symphysis, a 6-Fr single-J ureteral stent was placed in the ureter. A conduit was irrigated using a 22-Fr catheter to minimize spillage of bowel contents. A single-J ureteral stent attached to the ureter was passed through two enterotomies in the proximal conduit and 12-mm assistant port. Ureteroenteric anastomosis was carried out using a continuous running suture method comprising two running 4-0 synthetic absorbable polyglyconate sutures. The first stitch was placed on the tip of spatulation of the ureter and IC. The second stitch was placed on the end opposite to the first stitch, and both stitches were tied. Then, the short tails of the two stitches were pulled by two laparoscopic graspers inserted from the port above the pubis and assistant port. The traction of the short tails of the two sutures could adjust the suture line parallel to a laparoscopic needle holder in the surgeon’s right hand, which makes carrying out running suture easier. The long tail of the second stitch was used for the anterior part of ureteroenteric anastomosis with three or four stitches using the running suture method, and it was tied to the first stitch. The posterior part of anastomosis was inverted by pulling the short tail of both stitches, and running suture was carried out in the same manner. A 3–4-cm skin incision was made at the supraumbilical site, and the specimen was removed. As for extracorporeal IC, a 5–7-cm skin incision was made at the infraumbilical site. The isolated ileum for conduit was washed outside the incision, and ureteroenteric anastomosis was carried out using a continuous running suture method. Perioperative data were compared between the intracorporeal and extracorporeal IC groups using propensity score matching. The primary study end-point was postoperative early (within 90 days) complication rates. In total, 28 matched pairs were evaluated. No significant difference in preoperative data was found (Table S1). The operative time for intracorporeal IC diversion and ureteroenteric anastomosis was 187 31 and 74 17 min, respectively. The median total operative time in intracorporeal groups was significantly longer, but the total median blood loss significantly decreased compared with that in extracorporeal groups (560 vs 1165 mL, respectively, P < 0.001). Importantly, the postoperative early complication rate in the intracorporeal group was significantly lower than that in the extracorporeal group (39% vs 71%, P = 0.016; Table S2). Similarly, the postoperative early major complication rate (Clavien grade 3–5) tended to decrease in the intracorporeal group (7% vs 29%, respectively, P = 0.036). The details are summarized in Table S3. As for postoperative recovery of bowel function, the mean days to regular oral food intake were 4 and 7 days in the intracorporeal and extracorporeal groups, respectively (P < 0.001). The Kaplan–Meier method showed no significant difference in ureteroenteric strictures between both groups (P = 0.60; Fig. S1). Urological Notes