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Background: Telomeres are specific structures located at the ends of chromosomes that help maintain chromosome stability. In most tissues, telomeres become shorter as cells divide, a phenomenon thought to be associated with limitations on normal cell proliferation. Almost all types of cancer cells, including bladder cancer cells, express the enzyme telomerase, which can maintain or extend telomere length. Purpose: We examined telomerase activity in tumor specimens from a cohort of patients with bladder cancer and determined whether telomerase could be detected in exfoliated cancer cells present in urine from these patients. Methods: Spontaneously voided urine specimens and bladder-washing fluids (obtained by propelling normal saline into the bladder through a catheter and then withdrawing the liquid contents) were taken from 45 patients before they underwent surgery. Telomerase activity was examined by means of the TRAP (telomeric repeat amplification protocol) assay on extracts of tumor samples from 42 patients and extracts of exfoliated cells in urine and bladder-washing fluid from 42 and 43 patients, respectively. Standard cytologic examination (Pap staining) of urine specimens was also used to detect exfoliated cancer cells. Results: Telomerase activity was found in 41 (98%; 95% confidence interval [CI] = 87%-100%) of the 42 tumor samples examined. In contrast, it was not detected in normal bladder tissue from two autopsied individuals who were free of bladder cancer and five of six individuals who had bladder cancer. Telomerase was detected in exfoliated cells in 23 (55%; 95% CI = 39%-70%) of the 42 spontaneously voided urine specimens and in 36 (84%; 95% CI = 69%-93%) of the 43 bladder-washing fluids examined. Considering voided urine specimens and bladder-washing fluids together, telomerase was detected in exfoliated cells from 40 (89%; 95% CI = 76%-96%) of the 45 patients. Telomerase activity was not detected in bladder-washing fluids from 12 cancer-free individuals. Cancer cells were detected by means of standard cytologic examination in the urine of 19 (42%; 95% CI = 28%-58%) of the 45 patients. Urine cytologic examination detected cancer cells in one (8%; 95% CI = 0%-38%) of 12 patients with grade 1 tumors and in 13 (46%; 95% CI = 28%-66%) of 28 patients with grade 2 tumors. In contrast, telomerase activity was detected in exfoliated cells (in voided urine or bladder-washing fluids) from nine (75%; 95% CI = 43%-95%)-of 12 patients with grade 1 tumors and from 27 (96%; 95% CI = 82%-100%) of 28 patients with grade 2 tumors. Conclusion and Implication: Telomerase activity can be detected in exfoliated cells in urine from patients with bladder cancer, and measurement of this activity appears to be more sensitive in detecting the presence of cancer than standard urine cytologic examination. These findings suggest that measuring telomerase activity in exfoliated cells would be useful in the diagnosis and follow-up of patients with bladder cancer, a possibility that warrants further study. [J Natl Cancer Inst 1997;89:724-30]

Background and Purpose: Whether urethral injection of anesthetic and lubricating gel prior to outpatient flexible cystoscopy is worthwhile with regard to pain tolerance has been investigated only in a parallel randomized study. A crossover study was thus designed for further elucidation.Patients and Methods: Each of 33 male patients underwent three flexible cystoscopic examinations with intraurethral instillation of 11 mL of cold anesthetic gel (group 1), plain lubricating gel (group 2), or no gel (group 3). In every examination, 2% lidocaine gel was applied to the cystoscope. Although the cystoscopy was performed by two urologic surgeons, each patient underwent the three consecutive examinations with the same urologist. All the patients separately recorded pain levels during gel instillation, cystoscope insertion, and intravesical observation on a 100-mm visual analog scale after every cystoscopy.Results: From the median scores, the degree of pain resulting from gel injection was 77.0% and 98.0% of those for cystoscope insertion and intravesical observation, respectively. For each group stratified by anesthetic method, there was no significant difference in the pain score during either cystoscope insertion or intravesical observation.Conclusion: The pain caused by intraurethral gel instillation is significant compared with that from cystoscope insertion and intravesical observation. Anesthetic gel instillation appears to have no significant advantage over anesthesia-free flexible cystoscopic examinations.

Transforming growth factor-beta 1 (TGF-β1) plays a significant role in regulating the proliferation and apoptosis of prostate epithelial and stromal cells. We explored the association between the T (Leu) to C (Pro) polymorphism at codon10 of the TGF-β1 gene (TGFB1) and the risk of prostate cancer (PCa) or benign prostatic hyperplasia (BPH) in 351 PCa patients, 221 BPH patients and 303 male controls in Japan. There were significant differences in the CC versus TC + TT genotype distribution between PCa patients and male controls (P = 0.008), and between BPH patients and male controls (P = 0.041). Males with the TC or TT genotype had a 1.62-fold increased risk of PCa [95% confidence interval (95% CI) = 1.14–2.30, P = 0.007] and a 1.51-fold increased risk of BPH (95% CI = 1.02–2.24, P = 0.041) compared with those with the CC genotype, therefore suggesting the dominant effect of the TGFB1 T allele on development of PCa and BPH. There were no significant differences in the TGFB1 genotype distribution between different groups of tumor grades and stages in the PCa patients and no significant differences when PCa patients were stratified by the age of onset. The results suggest that the codon10 polymorphism in TGFB1 may have a significant influence on the development of PCa and BPH, therefore underscoring the importance of the TGF pathway in the development of these prostatic diseases. However, it appeared to have no impact on the disease status or age of onset of PCa.

The SRD5A2gene codes the steroid 5-reductase type II, a critical mediator of androgen action, and the V89Land A49Tpolymorphisms of this gene may be associated with a distinct enzyme activity. We explored the association among these polymorphisms and the risk of prostate cancer or benign prostatic hyperplasia (BPH) in a Japanese population.

CCND1 mRNA is alternatively spliced to produce 2 transcripts, and the splicing pattern may be modulated by a frequent A870G single-nucleotide polymorphism within the conserved splice donor site of exon 4. Several studies have suggested a significant association between the CCND1 genotype and onset or progression of various cancers. To investigate the correlation of the polymorphism with genetic susceptibility to PCa and its disease status, we examined the polymorphism in 214 cases of PCa, 234 cases of BPH and 254 male controls. The CCND1 A allele was more frequently observed in the PCa group (p = 0.015) and the BPH group (p = 0.018) than the control group. Men with the AA genotype had an increased risk of PCa (aOR = 1.93, 95% CI 1.13–3.30, p = 0.016) and BPH (aOR = 1.84, 95% CI 1.09–3.09, p = 0.023) compared to those with the GG genotype. No significant association was observed when men with the AG genotype were compared to those with the GG genotype (PCa: aOR = 1.00, 95% CI 0.65–1.54, BPH: aOR = 0.91, 95% CI 0.60–1.39). The risk of PCa associated with the AA genotype appeared to be stronger in men aged 73 years or younger (aOR = 2.89, 95% CI 1.38–6.01, p = 0.005), whereas no association was found in men older than 73 years (aOR = 1.02, 95% CI 0.44–2.34). No significant difference in genotype frequency was found among patients with low-, intermediate- and high-grade tumors (p = 0.730) or between patients with localized and metastatic PCa (p = 0.679). However, in patients with high-grade or metastatic PCa, a significantly increased risk associated with the AA genotype compared to controls was observed, while no significant results were found in those with low/intermediate or localized PCa. The A allele of the CCND1 A870G polymorphism was recessively associated with susceptibility to PCa and BPH in a Japanese population, giving a 2-fold increased risk of PCa and BPH in men with the AA genotype compared to those with the GG genotype. Although the risk of PCa associated with the AA genotype appeared to contribute especially to men aged 73 years or younger and the A allele may be associated with disease status of PCa, these conjectures require validation in future studies on a larger number of subjects. © 2002 Wiley-Liss, Inc.

Hematuria is absent in a significant proportion of patients with acute ureterolithiasis. We determined whether time from pain onset has any impact on the sensitivity of hematuria tests in the diagnosis of ureterolithiasis.

The progression of a malignant tumour is understood to be the result of the accumulation of multiple genetic aberrations. As up to 14% of organ‐confined renal cell carcinomas will recur after surgery, tumour clones with metastatic potential must already be present in some of these localized tumours. The association of 14q LOH with high‐grade tumours and advanced tumour stage suggests an important role for the gene in tumour progression. Chromosome 14q LOH has been analysed in microdissected specimens from 130 organ‐confined (UICC TNM stage 1 and 2) clear cell renal cell carcinomas using three microsatellite markers (D14S588, D14S617, GATA136B01). Tumours were classified as 14q LOH or not on the basis of LOH at one or more of the markers. The allelic imbalance ratio was used to determine both LOH and LOH proportion and the association between LOH and mortality, tumour size, histological grade and growth kinetics, measured by quantification of nucleolar organizer regions, was analysed. 14q LOH was present in 35.4% of informative cases at marker D14S588, 24.4% at D14S617, 36.4% at GATA136B01 and 39.5% for any one of the three markers. The mean 14q LOH proportion was 0.24 (range 0.009–0.80). LOH proportion correlated significantly with tumour size, AgNOR score and histological grade. It was also significantly associated with disease‐specific mortality; (hazard ratio 1.22; 95% CI 1.02–1.45; p= 0.039). LOH proportion did not remain significant after adjusting for tumour size (hazard ratio 0.98; 95% CI 0.76–1.27; p= 0.90). These results indicate that the proportion of cells with 14q LOH in the tumour is associated with tumour aggressiveness; while this is not an independent predictor of survival, it may have some utility as a marker of latent metastatic potential. Copyright © 2002 John Wiley & Sons, Ltd.

The amplification and overexpression of the human epidermal growth factor receptor 2 gene HER-2 (also known as c-erb-B2 or neu) have been shown to be associated with bladder cancer and its progression. Recent studies indicated an association between the Ile to Val polymorphism at codon 655 of HER-2 and susceptibility to breast cancer. To investigate the correlation between the Ile/Val polymorphism and the susceptibility and progression of bladder cancer, we analyzed the polymorphism in 232 patients with transitional cell carcinoma of the bladder and 408 normal controls. The frequencies of the Ile/Ile, Ile/Val and Val/Val genotype were 75.9%, 21.6% and 2.6%, respectively, in patients with bladder cancer and 75.7%, 23.0% and 1.2%, respectively, in controls. Statistical analyses of the genotype prevalence showed no significant difference between bladder cancer patients and normal controls (p = 0.419). Moreover, no significant differences in the genotype prevalence were observed when the patients were stratified according to the tumor grade, stage and smoking habits. When the Ile/Ile genotype was compared to the Ile/Val and Val/Val genotypes, a significant difference was found only between the patients with tumor stage Ta and those with T1-4 (age, gender and smoking habits-adjusted odds ratio = 2.13, 95% confidence interval = 1.09–4.15, p = 0.027). When the Ile/Ile + Ile/Val genotypes compared to the Val/Val genotype, no significant findings were observed. These results suggested that the HER-2 polymorphism at codon 655 is unlikely to be associated with the onset of bladder cancer. Furthermore, the findings suggest no association between this polymorphism and the disease progression in bladder cancer, although the possibility remains that the Ile/Ile genotype may be related to an increased risk of disease progression. © 2001 Wiley-Liss, Inc.

Cyclin D1 is believed to play an important role in the genesis and/or progression of transitional cell cancer (TCC) of the urinary bladder. Cyclin D1 gene (CCND1) mRNA is alternatively spliced to produce two transcripts, and the splicing pattern may be modulated by a G to A single nucleotide polymorphism within the splice donor site of exon 4. This study was conducted to explore the association between the polymorphism and the susceptibility to and disease status of TCC of the bladder in 222 cases and 317 native Japanese controls. The relationship between the CCND1 polymorphism and the mRNA splicing pattern in TCC cells was evaluated by semi-quantitative reverse-transcription PCR. The CCND1 A allele was more frequently observed in the TCC group than the control group (P = 0.032) with a significant difference in the genotype frequency between the two groups (P = 0.029). The AA genotype was associated with a significantly higher risk of TCC compared with the AG+GG genotypes (adjusted odds ratio (aOR) = 1.76, 95% confidence interval (CI) = 1.09-2.84, P = 0.022). This association was observed more significantly in nonsmoking cases (aOR = 2.53; 95% CI = 1.28-4.51, P = 0.008). Looking at tumor grade, the presence of the A allele was associated with higher grade (= grade 3) tumors with a gene dosage effect (aOR = 1.77, CI = 1.16-2.69, P = 0.008). In tumor stage, although not significant, the AA + AG genotypes tended to be more frequently observed in cases with T1-4 tumors than those with Ta tumors (aOR = 1.94, 95% CI = 0.98-3.82, P = 0.057). The genotype seemed to influence the two alternatively spliced forms of the CCND1 mRNA because the ratio of the CCND1 transcript-b/transcript-a was significantly higher in cases with the AA genotype compared with those with the AG + GG genotypes. These data suggest that the CCND1 variant A allele may be associated with an increased risk of TCC of the bladder, especially in men without a history of smoking, and it may also have an effect on its disease status.

We recorded auditory evoked magnetic fields (AEFs) by presenting pure tone bursts once every 4000 ms in 11 patients with a brain lesion in or in the vicinity of the auditory cortex. AEFs on the damaged side revealed several enhanced deflections in late-latency AEFs (slow AEFs), peaking at ∼320 (D1), 1030 (D2) and 1600 (D3) ms post-stimulus in eight patients. All the dipoles of slow AEFs were concentrated in the superior temporal regions which were not involved by brain lesions. D1, D2 and D3 dipoles were uniformly upward, downward and upward, respectively. The dipole moment varied from 12 to 122 nAm and had no consistent relationship with latency. This is the first report describing slow AEFs in cases with temporal lobe lesions.

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.

Trinucleotide repeats are polymorphic in normal individuals. CAG repeats in the X-linked androgen receptor gene were counted by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF-MS). A region of approximately two hundred base-pairs containing the repeats was amplified by polymerase chain reaction, then measured after a simple purification procedure. The single-charged molecular ion species was detected using 0.1 pmol of DNA sample and the number of repeats was determined from the molecular mass. The results indicated that MALDI/TOF-MS is a high-throughput alternative to polyacrylamide gel electrophoresis for precise determination of polymorphic trinucleotide repeats. Copyright © 1999 John Wiley & Sons, Ltd.

We report a case of dysplastic gangliocytoma of the cerebellum (Lhermitte-Duclos disease, LDD). The patient also had cutaneous and mucosal hamartomas, adenomatous goiter, bilateral breast tumors, and gastrointestinal polyposis, indicating the diagnosis of Cowden disease (CD), the familial hamartoma syndrome. This was a rare sporadic case without any family history of CD, though CD is considered to be an autosomal dominant hereditary disease. Based on a thorough review of the previously reported cases, it is reasonable to consider that CD is inherited in autosomal dominant fashion through a CD gene (PTEN) containing a germline mutation, and that the occurrence of LDD is predicated on an additional somatic hit on the remaining normal CD allele or another unknown gene.

Regional cerebral blood flow (rCBF) and its reactivity to acetazolamide were analyzed in a series of 15 cases of childhood moyamoya disease, using the xenon-133 inhalation method and single photon emission CT (SPECT). Most of the patients had normal mean hemispheric cerebral blood flow (mCBF), comparable to that of age-matched healthy children. However, they had abnormal rCBF distribution and disturbed reactivity to acetazolamide in the territory of the internal carotid artery, especially in the frontal lobe. Surgical revascularization for the anterior circulation, which consisted of STA-MCA anastomosis and indirect synangiosis, dramatically improved theses cerebral hemodynamics in the anterior circulation, including the frontal lobe, and reactivity to acetazolamide in the whole brain. These results suggest that surgical revascularization for the anterior circulation should be carried out in order to improve cerebral hemodynamics as widely as possible, especially in the frontal lobe, and may improve cerebral perfusion reserve in the posterior circulation.

S and F1C fimbrial adhesins often expressed by uropathogenic Escherichia coliare genetically homologous. A multiply primed polymerase chain reaction (PCR) was developed for discriminating the S (sfa) and F1C (foc) fimbrial operons. A total of 270 uropathogenic E. colistrains and 80 fecal isolates were examined. PCR specifically detected the sfaand focalleles in 105 (93%) of 113 sfa/foc+strains by DNA hybridization. Furthermore, 87% of sfa+uropathogenic E. colisimultaneously possessed the genes encoding the class III P fimbrial adhesin (prsGJ96), α‐hemolysin and cytotoxic necrotizing factor 1. Statistical analysis showed the class II P fimbrial adhesin (papGIA2) and F1C fimbria to be associated with high relative virulence in pyelonephritis and cystitis, respectively. The multiply primed PCR developed should be useful for assessing the contribution of the S and F1C fimbriae in the pathogenesis of urinary tract infections.

“Re-build-up” phenomenon, induced by hyperventilation, is a characteristic finding on electroencephalogram (EEG) in children with moyamoya disease, and suggests close correlation with cerebral ischemic attack. Its mechanism, however, remains obscure. In this study, we examined the relationship between the cerebral hemodynamics and this phenomenon before and after vascular reconstructive surgery. Regional cerebral blood flow and its reactivity to acetazolamide were analyzed using the xenon-133 inhalation method and single photon emission computed tomography in six children with moyamoya disease. These results were compared with the EEG. Regional vasoreactivity to acetazolamide was significantly lower in “re-build-up”-positive regions than in “re-build-up”-negative regions. Postoperative studies revealed a significant improvement of reactivity in the regions where the “re-build-up” phenomenon disappeared after surgery. These results suggest that the “re-build-up” phenomenon represents a focal reduction of the cerebral perfusion reserve in moyamoya disease.

Abstract: Thirteen pediatric patients (ages 4–13 years) who underwent surgical treatment were examined regarding their rCBF in the preoperative periods. The postoperative rCBF was measured 39 times in these 13 patients. Thirteen healthy normal subjects (ages from 6 to 21 years) were also examined. The rCBF in the operculum and in the frontal, parietal, and occipital lobes was measured with 133Xe inhalation method and single photon emission computed tomography. In the parietal and occipital lobes, the preoperative rCBF had a negative and significant correlation with their ages, but not in the operculum or frontal lobe. However, subsequent to the surgical treatment, the rCBF increased significantly in the patients 5 years old or less, and then post-operative rCBF values had significant negative correlations with age in each region.

BackgroundEscherichia coliis the most frequent pathogen in both acute bacterial prostatitis and acute uncomplicated urinary infections. To assess the virulence profiles of E. coliin acute prostatitis, the serotypes and virulence factor (VF) genotypes were determined.

PurposePrevious studies indicated that acute pyelonephritis in infants is initiated by the dominance of uropathogenic strains in fecal flora. Such pathogenic evidence, however, is still lacking for adult women. In this study, the validity of a fecal-perineal-urethral hypothesis in acute uncomplicated pyelonephritis of adult women was assessed at a genetic level.Materials and MethodsA total of 1,200 Escherichia coli isolates from the urine and rectal swab of 12 adult women with acute uncomplicated pyelonephritis were examined. The clonality of the urinary and fecal isolates was evaluated by genotyping of 6 urovirulence determinants and pulsed-field gel electrophoresis. Furthermore, urovirulence genotypes were examined in E. coli isolates from the rectal swab of 30 normal healthy women (mean 26.7 isolates per person).ResultsThe E. coli strains causing pyelonephritis were present in the rectal swab in 10 of 12 patients and were a predominant fecal clone in 9 cases. Also, P-fimbriated strains dominated in the fecal flora in 10 of 30 normal healthy women.ConclusionsThe clonal identity of the urinary and fecal strains in acute pyelonephritis clearly supports the fecal-perineal-urethral hypothesis.

In malignant tumors, telomerase reactivation plays an important role in the acquisition of cellular immortality. We evaluated the telomerase activity in renal cell carcinomas (RCCs) with special reference to their clinicopathologic features.

The acute effects on the sleep-wakefulness cycle of 2-octyl-?-bromo-acetoacetate (?-Br), an organic bromine compound which occurs naturally in the brain and cerebrospinal fluid (CSF) of mammals, were studied in the encéphale isolé cat preparation under artificial respiration without fixation.

S and F1C fimbrial adhesins often expressed by uropathogenic Escherichia coli are genetically homologous. A multiply primed polymerase chain reaction (PCR) was developed for discriminating the S (sfa) and F1C (foc) fimbrial operons. A total of 270 uropathogenic E. coli strains and 80 fecal isolates were examined. PCR specifically detected the sfa and foc alleles in 105 (93%) of 113 sfa/foc+ strains by DNA hybridization. Furthermore, 87% of sfa+ uropathogenic E. coli simultaneously possessed the genes encoding the class III P fimbrial adhesin (prsGJ96), a-hemolysin and cytotoxic necrotizing factor 1. Statistical analysis showed the class II P fimbrial adhesin (papGIA2) and F1C fimbria to be associated with high relative virulence in pyelonephritis and cystitis, respectively. The multiply primed PCR developed should be useful for assessing the contribution of the S and F1C fimbriae in the pathogenesis of urinary tract infections.