Fluorescence in situ hybridization evaluation of c-erbB-2 gene amplification and chromosomal anomalies in bladder cancer.

Oncogene amplification and chromosomal anomalies are found in many solid tumors and are often associated with aggressiveness of cancer. We evaluated the frequency and the role of c-erbB-2 gene amplification, relative increase in c-erbB-2 gene copy number, and gain of chromosome 17 in bladder cancer. A total of 29 bladder cancer specimens were examined using fluorescence in situ hybridization (FISH). Dual labeling hybridization with a directly labeled centromere probe for chromosome 17 together with a probe for the c-erbB-2 locus was performed. c-erbB-2 gene amplification was found in 3.4% (1 of 29) of specimens. Relative increase in c-erbB-2 gene copy number was found in 41.4% (12 of 29) of specimens and was significantly associated with tumor grade (P = 0.044 by Fisher's exact test). Gain of chromosome 17 was identified in 65.5% (19 of 29) of specimens and was significantly associated with tumor grade (P = 0.002 by Fisher's exact test) and tumor stage (P = 0.003 by Fisher's exact test). Our results suggest that c-erbB-2 gene amplification, relative increase in c-erbB-2 gene copy number, and gain of chromosome 17 may play important roles in the development and progression of bladder cancers. Moreover, the use of c-erbB-2 amplification, relative increase in c-erbB-2 gene copy number, and gain of chromosome 17 using FISH, together with tumor grade and stage, may provide a more useful clinical indicator in bladder cancer.

[Diagnosis of prostate capsular invasion by pelvic magnetic resonance imaging and serum level of prostate specific antigen].

Transrectal ultrasonography (TRUS), computed tomography (CT), and magnetic resonance imaging (MRI) are employed to diagnose the clinical stage of prostate cancer. However, several cases are diagnosed as pathological stage pT3 after total prostatectomy. We investigated the accuracy of the evaluation of pathologic capsular penetration by preoperative pelvic MRI and preoperative serum PSA level and capsular penetration. The diagnostic accuracy of capsular penetration by MRI was 63.3%. On the other hand, the diagnostic accuracy of capsular penetration by preoperative PSA was 89.7% when its cut off value was 17 ng/ml. We conclude that preoperative serum PSA level could be more useful to diagnose accurately stage of prostate cancer than pelvic MRI.

[Clinical outcome and analysis of radical prostatectomy in clinical stage B and C prostate cancer].

Between 1989 and 1999, 40 patients treated with radical prostatectomy for clinical stage B and C prostate cancer were analyzed. Prostate-specific antigen (PSA) failure after radical prostatectomy occurred in four patients, and one of them died of clinical recurrence of prostate cancer. Cause-specific survival at 5 years was 91.7% and PSA failure-free rate at 5 years was 76.8%. Staging accuracy of CT and MRI image was not satisfactory. In 41.7% patients, extracapsular extension can not be determined. Preoperative serum PSA levels of pathologically organ-confined disease (OCD) patients were significantly lower than those of pathologically non-OCD patients. Further analysis indicated that preoperative serum PSA levels of greater than 20.1 ng/ml are useful predictors for pathologically non-OCD.

Reclassification of Pichia scaptomyzae and Pichia galeiformis.

Based on the base composition of nuclear DNA and DNA/DNA hybridization, Pichia galeiformis IFO 10718T was reclassified as a synonym of Pichia manshurica, and Pichia scaptomyzae IFO 1073 1T was confirmed to be a synonym of Pichia membranifaciens. Comparison of 18S rRNA gene sequences indicated that IFO 10731T (P. scaptomyzae) is identical to P. membranifaciens IFO 10215T and IFO 10725, and IFO 10718T (P. galeiformis) is identical to P. manshurica IFO 10726T. These data were consistent with the view that P scaptomyzae and P membranifaciens should be conspecific, as should P. galeiformis and P manshurica. Variation among 26S rRNA gene domain D1/D2 sequences from three P membranifaciens strains indicated that this species encompasses a genetically heterogeneous population.

Three new species of Saccharomyces sensu lato van der Walt from Yaku Island in Japan: Saccharomyces naganishii sp. nov., Saccharomyces humaticus sp. nov. and Saccharomyces yakushimaensis sp. nov.

Three new yeast species were isolated from soil and partially decayed leaves in Yaku Island and Iriomote Island in the Nansei Islands of Japan. Based on DNA hybridization and physiological characters, these represent novel taxa. These are designated Saccharomyces naganishii sp. nov. (type strain IFO 10181T = CBS 8797T), Saccharomyces humaticus sp. nov. (type strain IFO 10673T = CBS 8893T) and Saccharomyces yakushimaensis sp. nov. (type strain IFO 1889T = CBS 8894T). The phylogenetic relationships of the three new species with members of other ascomycetous genera (e.g. Kluyveromyces, Saccharomyces, Torulaspora and Zygosaccharomyces) were estimated by 18S rDNA gene sequence analysis.

Fluorescence in situ hybridization evaluation of c-myc and androgen receptor gene amplification and chromosomal anomalies in prostate cancer in Japanese patients.

BACKGROUND: Oncogene amplification and chromosomal anomalies are found in many solid tumors and are often associated with aggressiveness of cancer. We evaluated the frequency and the association of c-myc and androgen receptor (AR) gene amplification and gain of chromosome 8 or X in prostate cancer in Japanese patients. METHODS: We examined a total of 42 prostate cancer specimens, using fluorescence in situ hybridization (FISH). Dual-labeling hybridization with a directly labeled centromere probe for chromosome 8 or X together with a probe for the c-myc or AR locus was performed. RESULTS: Gain of chromosome 8 was identified in 54.8% of specimens and was associated with Gleason sum and nuclear anaplasia in untreated prostate cancers. c-myc gene amplification was found in 14.3% of specimens. Gain of chromosome X was identified in 42.9% of specimens. AR gene amplification was detected in 0 of 37 untreated prostate cancers, but in 1 of 5 hormone-refractory prostate cancers. CONCLUSIONS: Our results suggest that c-myc and AR gene amplification and gain of chromosome 8 or X may be associated with the development and progression of prostate cancers. These results obtained in Japanese cases are consistent with the results observed in prostate cancer in Western countries.

Reclassification of Pichia membranifaciens sensu Kurtzman.

Forty strains which were stocked as Pichia membranifaciens sensu Kurtzman and 9 strains stocked as Candida valida, anamorphs of P. membranifaciens, in the Institute for Fermentation, Osaka (IFO) were reclassified based on the data of base composition of nuclear DNA, DNA/DNA hybridization, coenzyme Q system, electrophoretic karyotype, and base sequence of 18S rDNA. P. membranifaciens complex was assigned into 3 groups: (I) P. membranifaciens group, including 25 strains with high DNA homologies to the type strain of P. membranifaciens (72-98%); (II) P. manshurica group, including 18 strains with high DNA homology of 79-95% to the type strain of P. manshurica; and a group including the remaining 6 strains, which had low DNA homology to the above two species. GC content was 42.9-45.3 mol% for the P. membranifaciens group, 40.0-42.0 mol% for the P. manshurica group, and 27.2-44.7 mol% for the remaining group. All three groups had ubiquinone Co Q-7. Of the 6 anomalous strains, IFO 0162 was identified as Pichia deserticola, and IFO 0839 and IFO 0840 were identified as Issatchenkia occidentalis; but IFO 0842, IFO 0843, and IFO 1788 were thought to be unknown strains.

[Study of free to total prostate specific antigen ratio in the detection of patients with prostate cancer].

BACKGROUND: We investigated the clinical usefulness of free to total serum prostate specific antigen (PSA) ratio (F/T ratio) in order to improve the specificity of total PSA measurement for detecting prostate cancer. METHOD: In this study 129 patients with total PSA level 4-20 ng/ml underwent transrectal ultrasound guided sextant biopsy. Serum samples were assessed for total PSA, free PSA and the F/T ratio calculated. All patients were pathologically diagnosed as benign prostatic hyperplasia or prostate cancer. RESULTS: Of 129 patients 21 had prostate carcinoma (PCa) and 108 had benign prostatic hyperplasia (BPH) from the results of prostate biopsies. The mean of total PSA were not significantly different between men with PCa and with BPH. The mean of free PSA for PCa was significantly lower than that for BPH (p = 0.043). Furthermore, the mean of F/T ratio was significantly different between PCa and BPH group (p = 0.0014). The F/T ratio had a higher specificity than total PSA at all levels of sensitivity in detecting prostate cancers. Sensitivity, specificity and accuracy for cancer detection at a cut off 0.12 was 90.4%, 51.8% and 58.1%, respectively. Also, free PSA was as useful as F/T ratio for cancer detection when analyzed in receiver operating characteristic curves analysis. When determined the cut off number of free PSA at 0.78 ng/ml, the sensitivity, specificity and accuracy for cancer detection were 61.9%, 66.7% and 65.9%, respectively. CONCLUSION: This study indicated that the F/T ratio and free PSA could improve the specificity without impairing the sensitivity for detecting PCa in patients with 4-20 ng/ml of total PSA.

A new yeast genus, Tetrapisispora gen. nov.: Tetrapisispora iriomotensis sp. nov., Tetrapisispora nanseiensis sp. nov. and Tetrapisispora arboricola sp. nov., from the Nansei Islands, and reclassification of Kluyveromyces phaffii (van der Walt) van der Walt as Tetrapisispora phaffii comb. nov.

Seven strains of three new yeast species were isolated from soil, flowers and leaves in the Nansei Islands, Japan. These isolates most closely resembled Kluyveromyces phaffii in physiological characteristics and nuclear DNA base composition (30-32 mol% G + C), but on the basis of DNA-DNA hybridization and electrophoretic karyotyping they were categorized into three new species different from K. phaffii. Phylogenetic analysis using 18S rRNA gene sequences showed that the three new species and K. phaffii were highly related to one another and phylogenetically separate from the members of other species. On the basis of phylogeny and physiological characters, it is proposed that the three new species represent novel taxa and should be designated Tetrapisispora iriomotensis gen. nov., sp. nov. (type strain IFO 10929T), Tetrapisispora nanseiensis gen. nov., sp. nov. (type strain IFO 10899T) and Tetrapisispora arboricola gen. nov., sp. nov. (type strain IFO 10925T), while Kluyveromyces phaffii becomes Tetrapisispora phaffii comb. nov.

The worst histologic elements predict prognosis in moderately-differentiated prostate cancer patients with androgen-withdrawal endocrine therapy.

OBJECTIVES: We evaluated the prognostic value of the worst histologic elements in 155 patients with moderately-differentiated prostate cancer (Gleason sum 5-7) treated with androgen-withdrawal endocrine therapy. METHODS: In individual cases, the proportion (%) of medullary/solid combined with columns-and-cords/trabecular (MED & C-C), regarded as the worst histologic elements, determined according to WHO-Mostofi's classification of histologic patterns, within the total adenocarcinoma area obtained from needle specimens was estimated. RESULTS: Patients with proportion of MED & C-C >5% had a significantly poorer prognosis than those with <5% (p < 0.05). A significant difference was also found in the survival curves of patients grouped by stage (p < 0.01). There was no significant difference among the patients with Gleason sum of 5, 6 and 7. Multivariate analysis showed that stage and MED & C-C had independent prognostic value. CONCLUSIONS: Proportion of MED & C-C may provide useful prognostic information about patients with moderately-differentiated, Gleason sum 5-7, prostate cancer treated with androgen-withdrawal endocrine therapy.

The phylogeny of the cactophilic yeasts based on the 18S ribosomal RNA gene sequences: the proposals of Phaffomyces antillensis and Starmera caribaea, new combinations.

The complete sequences of the 18S rRNA gene fragments of the type strains of the cactophilic yeast species, Pichia antillensis, Pichia caribaea, Phaffomyces opuntiae, Phaffomyces thermotolerans, Starmera amethionina var. amethionina, and Starmera amethionina var. pachycereana were determined and compared. The type strain of Phaffomyces opuntiae had two kinds of the 18S rRNA gene sequences of which base differences were counted to be 15 and of which the percent similarity was calculated to be 99.1. The type strains of P. antillensis, P. caribaea, and Starmera amethionina var. pachycereana had the Q-7 system. The phylogenetic analyses showed that the genera Phaffomyces and Starmera were monophyletic and distant from each other and from the other species examined of the ascogenous teleomorphic genera, and that P. antillensis and P. caribaea were included within the clusters of the genera Phaffomyces and Starmera, respectively. The two Pichia species were transferred to the genera Phaffomyces and Starmera as the new combinations, Phaffomyces antillensis and Starmera caribaea. The new family Phaffomycetaceae was proposed as the type genus Phaffomyces.

[Bilateral renal cell carcinoma in a patient with chronic renal failure: a case report].

We describe a case of bilateral renal cell carcinoma associated with chronic renal failure. A 49-year-old man was admitted to our hospital to initiate hemodialysis. He was found to have multiple renal cystic changes and a left renal mass by ultrasound. Computed tomographic (CT) scan showed a tumor 4 cm in diameter in the left kidney and another one 2 cm in diameter in the right kidney. Left nephrectomy was first performed and histopathological examination revealed renal cell carcinoma of mixed type and granular subtype. Six months after the operation, CT scan showed the mild growth of the tumor in the right kidney. Right nephrectomy was performed and histopathological examination revealed renal cell carcinoma of papillary type and granular subtype. The patient remains well on hemodialysis, with no evidence of recurrence or metastasis for 13 months after nephrectomy.

The phylogeny of species of the genus Issatchenkia Kudriavzev (Saccharomycetaceae) based on the partial sequences of 18S and 26S ribosomal RNAs.

The ten strains of Issatchenkia species were examined for their partial base sequences of 18S and 26S rRNAs. In the 18S rRNA partial base sequences (positions 1451-1618, 168 bases), the strains of the species of the genus Issatchenkia were found to be not uniform phylogenetically. The calculated base differences numbered 5-0. The strains of Issatchenkia species examined had 3-1 base differences with the type strain of Pichia membranaefaciens. Especially, the type strain of Issatchenkia orientalis, the type species of the genus Issatchenkia was found to be closely related phylogenetically to that of P. membranaefaciens. The calculated number of base differences was only one. The base sequences on the fingerprint segment were comprised of four bases (four kinds of AUAU, CCAU, AUAG, and ACAU), as found in P. membranaefaciens (ACAA). In the 26S rRNA partial base sequences, the calculated number of base differences was 8-0 (positions 1611-1835, 225 bases), and the calculated percent similarities were 61-80 (positions 493-622, 130 bases), within the genus Issatchenkia. Discussion was made phylogenetically and taxonomically, especially on the phylogenetic relationship between the type species of the genera Issatchenkia and Pichia and on a circumscription of the genus Issatchenkia.

Detection of heterogeneity of 18S rRNA inter-genes and mutation arising during PCR amplification.

Direct sequencing revealed sequence heterogeneity among ribosomal RNA gene (rDNA) operons, consisting of 8 base heterogeneous sites on the 18S rDNA of Galactomyces citri-aurantii IFO 10822, and 6 base heterogeneous sites in the same region on the 18S rDNA of G. citri-aurantii IFO 10821. Sequence analysis of the cloned 18S rRNA genes of 14 species (19 strains) of ascomycetous yeast-like fungi detected a total of 32 substitutions between two cloned sequences from each of 10 strains. Eight substitutions came from heterogeneity of G. citri-aurantii IFO 10822, and 24 substitutions were predicted to be due to misincorporation by the Taq DNA polymerase. A low frequency of random substitution, estimated to occur in PCR at approximately 1 in 2690 nucleotides, was detected; and transitions occurred 7 times more frequently than transversions.

The phylogeny of species of the genus Saccharomycopsis Schiönning (Saccharomycetaceae) based on the partial sequences of 18S and 26S ribosomal RNAs.

Eight strains of Saccharomycopsis species were examined for their partial base sequences of 18S and 26S rRNAs. The base sequences on the fingerprint segment showed three kinds of ACUAU, UCUAU, and AUAU. In the partial base sequences (positions 1451-1618, 168 bases) of 18S rRNA, all the strains of Saccharomycopsis species examined had 1-0 base difference except for the strains of Saccharomycopsis fibuligera (base differences, 5-4). In the partial base sequences on the positions 1611-1835 region (225 bases) of 26S rRNA, however, S'copsis vini had 15-11 base differences. The other species had 7-2 base differences. In the partial base sequences on the positions 493-622 region (130 bases) of 26S rRNA, the calculated percent similarities were 62-77. Discussion was made phylogenetically and taxonomically, especially on the separation of S'copsis fibuligera (identical to Endomyces fibuliger) from the genus Saccharomycopsis.

Phylogenetic relationships of species of the genus Saccharomyces Meyen ex Reess deduced from partial base sequences of 18S and 26S ribosomal RNAs (saccharomycetaceae).

In order to clarify the phylogenetic relationships among the yeast species classified in the genus Saccharomyces, partial base sequences of 18S and 26S ribosomal RNAs were determined for ten selected strains. The regions determined correspond to positions 1451 through 1618 of the 18S rRNA and positions 493 through 622 and 1611 through 1835 of the 26S rRNA in S. cerevisiae. Analyses of these partial base sequences suggested that the genus Saccharomyces is phylogenetically heterogeneous. Saccharomyces servazzii and S. unisporus showed identical or very similar sequences in all the three regions, and their phylogenetic distance from S. cerevisiae was large enough to introduce a genus independent of Saccharomyces. Saccharomyces kluyveri is also distant from all the other Saccharomyces species examined, and is likely to deserve a new genus. Estimated phylogenetic relationships between Saccharomyces and other genera characterized by the Q-6 system, such as species of Zygosaccharomyces. Torulaspora, Kluyveromyces, Arxiozyma, Pachytichospora, Nadsonia. Hanseniaspora, Kloeckeraspora, and Saccharomycodes, are also discussed.

Phylogenetic relationships of species of the genus Kluyveromyces van der Walt (saccharomycetaceae) deduced from the partial sequences of 18S and 26S ribosomal RNAs.

In order to clarify the phylogenetic relationships of the species classified in the genus Kluyveromyces (Saccharomycetaceae), three partial base sequences of 18S and 26S rRNAs of eighteen strains were determined. The regions determined of the strains corresponded to positions 1451 through 1618 (168 bases) of 18S rRNA and to positions 1611 through 1835 (225 bases) and 493 through 622 (130 bases) of a strain (IFO 2376) of Saccharomyces cerevisiae. The analyses of the partial base sequences suggested that the genus Kluyveromyces is phylogenetically heterogeneous, ranging from the strains that are quite close to the strain of S. cerevisiae to the strains that are distinct enough to be classified in genera separate from the genus Saccharomyces. From our sequence data, we concluded that the extent of the genus Kluyveromyces should be restricted to only one species, K. polysporus, the type species of the genus. Kluyveromyces phaffii was also distinct enough to deserve another genus. Kluyveromyces cellobiovorus was not close to any of the strains of Kluyveromyces species examined, and should be excluded from the genus. Most of the strains of the species examined were fairly close to the strain of S. cerevisiae.

[Extirpation of total urinary tract as a final treatment for asynchronous multicentric urothelial cancer: a case report].

A 48-year-old male patient underwent transurethral resection (TUR) for solitary bladder tumor in January 1985. Pathological finding of the specimen was non-invasive transitional cell carcinoma (TCC) G2 > G1. Following the operation, prophylactic intravesical instillation of antitumor agents, intrapelvic irradiation, and hyperthermia were done for the recurrences. In June 1987, left renal pelvic tumor revealed, and nephroureterectomy was performed. Histological examination of renal pelvic tumor showed TCC G1. Five months later, multiple bladder tumors and right renal pelvic tumor were detected. A total cystectomy, partial nephrectomy and nephrostomy were performed in February and March 1988. Pathology of the bladder and right kidney were TCC G3. One month after these operations, tumors recurred in the rest of the right kidney. Complete remission was achieved after systemic chemotherapy with methotrexate, vinblastine, doxorubicin and cisplatin (M-VAC). In January 1992, right renal pelvic tumor recurred again. After two cycles of M-VAC therapy, the rest of the right kidney was extirpated. Postoperative hemodialysis has been uneventful. He has lived without tumor recurrence for three years after the introduction of hemodialysis.

Should Petasospora BOIDIN et ABADIE (Saccharomycetaceae) be retained?--the phylogeny based on the partial sequences of 18S and 26S ribosomal RNAs.

The six strains of the Pichia species, once classified in the genus Petasospora, were examined for their 18S (positions 1451-1618, 168 bases) and 26S (positions 1611-1835, 225 bases and 493-622, 130 bases) rRNA partial base sequencings. In the 18S rRNA partial base sequencings, the type species of the genus Petasospora (Petasospora rhodanensis) was found to be closely related to Pichia anomala (identical to Hansenula anomala, type species of genus Hansenula) with base differences of two, but not to Pichia membranaefaciens, the type species of the genus Pichia, with base differences of ten. However, the species was not so closely related to P. anomala in the 26S rRNA partial base sequencings with base differences of twelve and 68 percent similarity. The genus Petasospora was known to be a very heterogeneous taxon phylogenetically with base differences of thirty-one to three and ninety-two to eight and with 40 to 83 percent similarities. The sequence data obtained here and the phenotypic features described previously indicate that the type species of the genus Petasospora (Pet. rhodanensis, identical to Pichia rhodanensis) is adequate to be accommodated temporarily in the genus Pichia, a heterogeneous taxon, until the precise taxonomic position of the type species is defined.

The phylogeny of Yamadazyma ohmeri (Etchells et Bell) Billon-Grand based on the partial sequences of 18S and 26S ribosomal RNAs: the proposal of Kodamaea gen. nov. (Saccharomycetaceae).

Two strains of Yamadazyma ohmeri were examined for their 18S and 26S rRNA partial base sequences. The two strains had identical or very similar partial base sequences to those of the type strain. All the three strains of Y. ohmeri were phylogenetically separated from any other accepted species of the genera Yamadazyma, Debaryomyces, Pichia, Williopsis, Metschnikowia, and Clavispora. Based on the sequence data obtained and the phenotypic characteristics, Kodamaea gen. nov. was proposed for the species.