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1.
Theor Appl Genet ; 126(7): 1851-9, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23604528

ABSTRACT

In order to determine the molecular basis of cytoplasmic male sterility (CMS) in alloplasmic lines of eggplant, the genomic structures and transcription patterns of mitochondrial ATP synthase subunit (atp) and cytochrome oxidase subunit (cox) genes were studied for wild and cultivated eggplants. Alloplasmic eggplant lines with cytoplasms of wild Solanum species showing either anther indehiscent type of CMS or non-pollen production type of CMS were studied with the cultivated eggplant Solanum melongena, used as a control. Southern hybridization of the mitochondrial genes indicated the difference between the two types of CMS and showed complete identity within each type. The cytoplasmic patterns of all wild species differed from that of the cultivated eggplant. Thus, the cytoplasm of the six wild eggplants and the one cultivated eggplant was classified into three groups. Male sterile plants of both types of CMS showed novel transcription patterns of atp1, whereas a different transcription pattern of cox2 was observed only in the anther indehiscent type. Based on these differences, we determined the DNA sequences of about a 4 kbp segment in the atp1 region. Although the coding and 3' flanking regions were almost identical among the cytoplasms, the 5' flanking region was completely different and novel open reading frames (orfs) were found for each of the CMS types and the cultivated eggplant. The cytoplasm of Solanum kurzii inducing the anther indehiscent type CMS had orf312, and those of Solanum aethiopicum and Solanum grandifolium of non-pollen production type CMS had orf218. The correspondence between the transcription patterns of these orfs and phenotypic expression of male sterility strongly suggests that these orfs are causal genes for each type of CMS.


Subject(s)
Genes, Mitochondrial , Genes, Plant , Plant Infertility/genetics , Solanum/genetics , Transcription, Genetic , Base Sequence , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA
2.
J Evol Biol ; 22(10): 2012-9, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19678867

ABSTRACT

In many gynodioecious species, sex determination involves both cytoplasmic male-sterility (CMS) genes and nuclear genes that restore male function. Differences in fitness among genotypes affect the dynamics of those genes, and thus that of gynodioecy. We used a molecular marker to discriminate between hermaphrodites with and without a CMS gene in gynodioecious Raphanus sativus. We compared fitness through female function among the three genotypes: females, hermaphrodites with the CMS gene and those without it. Although there was no significant difference among the genotypes in seed size, hermaphrodites without the CMS gene produced significantly more seeds, and seeds with a higher germination rate than the other genotypes, suggesting no fitness advantage for females and no benefit to bearing the CMS gene. Despite the lack of fitness advantage for females in the parameter values we estimated, a theoretical model of gynodioecy shows it can be maintained if restorer genes impose a cost paid in pollen production. In addition, we found that females invest more resources into female reproduction than hermaphrodites when they become larger. If environmental conditions enable females to grow larger this would facilitate the dynamics of CMS genes.


Subject(s)
Disorders of Sex Development , Ovule , Raphanus/physiology , Base Sequence , DNA Primers , Genotype , Models, Genetic , Polymerase Chain Reaction , Raphanus/genetics , Reproduction
3.
Transplant Proc ; 50(5): 1238-1242, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29880341

ABSTRACT

BACKGROUND: In this study we present our new surgical procedure, laparoendoscopic single-site surgery plus 1 for donor nephrectomy (LESS+1-DN), which shortens warm ischemic time (WIT) and improves surgical outcomes. METHODS: From January 2013 to February 2017, 15 patients who underwent LESS-DN and 41 patients who underwent LESS+1-DN at our institution were evaluated retrospectively. Patients were divided into 3 groups: group A, 15 cases of LESS-DN; group B, the first 15 patients who underwent LESS+1-DN; and group C, 26 patients who underwent subsequent LESS+1-DN. To reduce WIT, we clearly defined the roles of the surgeon and first assistant in the 26 subsequent LESS+1-DN cases. The surgeon dissected the renal pedicle and harvested the kidney graft using a recovery bag and the first assistant held the recovery bag. RESULTS: The mean operative time in group C (213.7 minutes) was significantly shorter than that in groups A (253.3 minutes) and B (253.8 minutes). The WIT in group C (195.2 seconds) was significantly shorter than that in groups A (389.8 seconds) and B (313.2 seconds). Open conversion was required in 1 case in group A. None of the donors required conversion to open surgery and no perioperative complications occurred in groups B and C. Linear regression analysis of the LESS+1-DN operative times and consecutive case numbers demonstrated a shallow learning curve (R2 = 0.392, P < .05). CONCLUSION: Our new procedure that divides the roles of the operator and the first assistant contributed significantly to a shortening of WIT. Dividing roles can facilitate a safer laparoscopic donor nephrectomy.


Subject(s)
Kidney Transplantation/methods , Nephrectomy/methods , Tissue and Organ Harvesting/methods , Warm Ischemia/methods , Adult , Aged , Conversion to Open Surgery/statistics & numerical data , Female , Humans , Laparoscopy/methods , Learning Curve , Length of Stay , Living Donors , Male , Middle Aged , Operative Time , Retrospective Studies
4.
J Natl Cancer Inst ; 89(10): 724-30, 1997 May 21.
Article in English | MEDLINE | ID: mdl-9168188

ABSTRACT

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.


Subject(s)
Carcinoma, Transitional Cell/enzymology , Telomerase/metabolism , Urinary Bladder Neoplasms/enzymology , Aged , Biomarkers , Carcinoma, Transitional Cell/diagnosis , Carcinoma, Transitional Cell/pathology , Female , Humans , Male , Middle Aged , Tumor Cells, Cultured , Urinary Bladder Neoplasms/diagnosis , Urinary Bladder Neoplasms/pathology
5.
Cancer Res ; 57(4): 742-6, 1997 Feb 15.
Article in English | MEDLINE | ID: mdl-9044854

ABSTRACT

A variety of malignancies have been linked to major histocompatibility complex genes, including the DRB1 alleles. The association of certain DRB1 antigens with renal cell carcinoma (RCC) has been both claimed and disclaimed. To determine whether HLA-DRB1 genotypes are associated with RCC, we used the modified PCR-RFLP method for the high-resolution HLA-DRB1 genotyping of 96 Japanese RCC patients. There were no significantly frequent HLA-DRB1 alleles, whereas the DRB1*0101 and *0405 alleles had significantly lower frequencies [P = 0.004, relative risk (RR) = 0.2 and P = 0.002, RR = 0.4) in the RCC patients than in the healthy Japanese controls (n = 1216). Moreover, patients with the HLA-DRB1 *0101 or *0405 allele tended to be in earlier stages and to have less aggressive tumors than patients with neither of these alleles. The corresponding serotyping subclassification, however, showed a significantly lower frequency only for DRB1-DR1 (P = 0.01, RR = 0.3). High-resolution genotyping is essential because the polymorphism of the peptide-binding domain of major histocompatibility complex class II molecules is more precisely determined by genotypes than serotypes. In addition, inherent technical difficulties and potential typing errors render serotyping inefficient. Our data suggest that HLA-DRB1*0101 and *0405 are protective alleles for both RCC development and tumor progression.


Subject(s)
Alleles , Carcinoma, Renal Cell/genetics , HLA-DR Antigens/genetics , Kidney Neoplasms/genetics , Adult , Aged , Aged, 80 and over , Carcinoma, Renal Cell/ethnology , Carcinoma, Renal Cell/pathology , Female , Genotype , HLA-DRB1 Chains , Humans , Kidney Neoplasms/ethnology , Kidney Neoplasms/pathology , Male , Middle Aged , Neoplasm Staging
6.
Cancer Res ; 51(10): 2521-3, 1991 May 15.
Article in English | MEDLINE | ID: mdl-1708695

ABSTRACT

Autologous immunoglobulin was detected on the cell surface of tumor cells freshly isolated from cancerous kidneys of patients with renal cell carcinoma by flow cytometry after staining with murine anti-human IgG monoclonal antibodies. Cells isolated in parallel from macroscopically normal regions of the tumorous kidneys were not specifically stained with the anti-human IgG reagents. In further studies, tumor cells were stained with an antibody to decay-accelerating factor (DAF), a known inhibitor of complement. Flow cytometry of these cells revealed that nearly all tumor cells expressed DAF, and that the intensity of staining with the anti-DAF monoclonal antibody correlated with the staining of cells with anti-IgG. The results suggest that tumor cells coated with autologous antibody may be resistant to complement-mediated cytotoxicity in vivo through the expression of high levels of DAF.


Subject(s)
Autoantibodies/analysis , Complement Inactivator Proteins/analysis , Kidney Neoplasms/immunology , Membrane Proteins/analysis , Receptors, Antigen, B-Cell/analysis , Antibodies, Monoclonal , CD55 Antigens , Flow Cytometry , Humans , Kidney Neoplasms/pathology , Lymphocytes/immunology , Lymphocytes/pathology
7.
Cancer Res ; 58(24): 5835-41, 1998 Dec 15.
Article in English | MEDLINE | ID: mdl-9865743

ABSTRACT

Recent molecular genetic studies have suggested that multifocal urothelial cancers are derived from an identical progenitor cell. However, the clonal origin of multifocal urothelial cancers of a low-grade superficial type has not been fully defined. Using microsatellite markers, we examined genetic alterations at 20 loci on eight chromosomal arms (2q, 4p, 4q, 8p, 9p, 9q, 11p, and 17p) in 87 metachronous and/or synchronous multifocal urothelial cancers, which included 84 low-grade superficial papillary tumors from 29 patients. Judging from the patterns of loss of heterozygosity, microsatellite shifts, and the subchromosomal partial deletion, multifocal tumors in at least 20 (80%) of the 25 evaluable patients were considered to be derived from a single progenitor cell, although the possibility remained that multifocal tumors in a small subset of patients might develop from distinct progenitor cells due to field cancerization. In 13 of the 20 patients, a chronological genetic analysis was available: genetic heterogeneity was detected in 3 (23%) patients, and an apparent accumulated pattern of genetic alterations was detected in only 1 (8%) patient. In the 20 patients with multifocal tumors of an identical clonal origin, discordant microsatellite alterations were observed, with significantly lower frequencies on chromosome 9 compared to those on the other chromosomes tested. The results indicate that most multifocal low-grade superficial urothelial cancers are genetically stable despite their incidence of frequent recurrence, and genetic divergence occurs in a subset of patients. This heterotopic spread and genetic divergence may occur long before the clinical manifestation of multiplicity from a single transformed cell. These data support the previous view that heterotopic spread of transformed progenitor cells and genetic divergence occur after chromosome 9 alterations in most of low-grade superficial urothelial cancers.


Subject(s)
Chromosome Aberrations , Clone Cells , Loss of Heterozygosity , Neoplasms, Multiple Primary/genetics , Urinary Bladder Neoplasms/genetics , Urologic Neoplasms/genetics , Genetic Heterogeneity , Humans , Microsatellite Repeats , Stem Cells
8.
Cancer Res ; 55(18): 4182-7, 1995 Sep 15.
Article in English | MEDLINE | ID: mdl-7664295

ABSTRACT

Mitogen-activated protein kinases (MAPKs) play a pivotal role in the mitogenic signal transduction pathway and are essential components of the MAPK cascade, which includes MEK (also known as MAP kinase kinase), Raf-1, and Ras. In this study, we examined whether constitutive activation of the MAPK cascade was associated with the carcinogenesis of human renal cell carcinomas in a series of 25 tumors and in corresponding normal kidneys. Constitutive activation of MAPKs in tumor tissue, as determined by the appearance of phosphorylated forms, was found in 12 cases (48%), and this activation was confirmed by a direct in vitro kinase assay of immunoprecipitate using myelin basic protein as the substrate. The phosphorylation of MEK and of Raf-1, as monitored by a mobility shift in SDS-PAGE, which is reportedly associated with the activation of these kinases, occurred in 9 of 18 cases (50%) and in 6 of 11 cases (55%) respectively. The activation of MAPKs was correlated with MEK activation (P = 0.0045) and with Raf-1 activation (P = 0.067). Furthermore, overexpression of MEK was found in 13 of 25 cases (52%) by Western blot analysis, and this overexpression was associated significantly with MAPK activation (P = 0.034). No mutations were noted in H-,K-, or N-ras genes by PCR direct sequencing in any of the 25 tumor samples. Of the patients studied, 8 of 18 (44%) stage pT2 patients and four of six (67%) stage pT3 patients showed MAPK activation. The single stage pT1 patient did not evidence MAPK activation. Furthermore, one of seven (14%) grade 1 patients, 9 of 13 (69%) grade 2 patients, and two of five (40%) grade 3 patients showed MAPK activation (grade 1 versus grades 2 and 3, P = 0.046). Our results suggest that constitutive activation of MAPKs may be associated with the carcinogenesis of human RCCs.


Subject(s)
Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Carcinoma, Renal Cell/enzymology , Kidney Neoplasms/enzymology , Amino Acid Sequence , Carcinoma, Renal Cell/pathology , Enzyme Activation , Genes, ras , Humans , Kidney Neoplasms/pathology , Mitogen-Activated Protein Kinase Kinases , Molecular Sequence Data , Protein Kinases/metabolism , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins/metabolism , Proto-Oncogene Proteins c-raf , Tumor Cells, Cultured
9.
Genetics ; 108(3): 681-95, 1984 Nov.
Article in English | MEDLINE | ID: mdl-17246239

ABSTRACT

The restriction fragment patterns of chloroplast DNAs of all M or modified M genome-carrying Aegilops species, and those of common wheat (Triticum aestivum), Ae. umbellulata and Ae. squarrosa as referants, have been analyzed using eight restriction endonucleases, BamHI, EcoRI, HindIII, KpnI, PstI, SalI, SmaI and XhoI. Nine distinctly different chloroplast genomes are evident, and the mutual relatedness among them is estimated based on the number of different restriction fragments. The results lead to the following conclusions. (1) Chloroplast genomes of three Comopyrum species, Ae. comosa, Ae. heldreichii and Ae. uniaristata, are more closely related with each other and are greatly different from those of the Amblyopyrum species, Ae. mutica, and of Ae. umbellulata and Ae. squarrosa. (2) Ae. crassa's chloroplast genome lies at the center of chloroplast genome diversification, whereas those of common wheat, Ae. squarrosa and Ae. uniaristata are three extreme forms lying far from the center. (3) Chloroplast genomes of three 4x species, Ae. biuncialis, Ae. columnaris and Ae. triaristata, arose from Ae. umbellulata, and that of a fourth 4x species, Ae. ventricosa , arose from Ae. squarrosa. The chloroplast origins of two other 4x species, Ae. ovata and Ae. crassa, remain unsolved. (4) The chloroplast genomes of two Ae. mutica strains are identical, even though their cytoplasms exert quite different effects on male fertility, heading date and growth vigor of common wheat.

10.
Genetics ; 129(3): 873-84, 1991 Nov.
Article in English | MEDLINE | ID: mdl-1752425

ABSTRACT

The nucleotide divergence of chloroplast DNAs around the hot spot region related to length mutation in Triticum (wheat) and Aegilops was analyzed. DNA sequences (ca. 4.5 kbp) of three chloroplast genome types of wheat complex were compared with one another and with the corresponding region of other grasses. The sequences region contained rbcL and psaI, two open reading frames, and a pseudogene, rpl23' (pseudogene for ribosomal protein L23) disrupted by AT-rich intergic spacer regions. The evolution of these genes in the closely related wheat complex is characterized by nonbiased nucleotide substitutions in terms of being synonymous/nonsynonymous, having A-T pressure transitions over transversions, and frequent changes at the third codon position, in contrast with the gene evolution among more distant plant groups where biased nucleotide substitutions have frequently occurred. The sequences of these genes had diverged almost in proportion to taxonomic distance. The sequence of the pseudogene rpl23' changed approximately two times faster than that of the coding region. Sequence comparison between the pseudogene and its protein-coding counterpart revealed different degrees of nucleotide homology in wheat, rice and maize, suggesting that the transposition timing of the pseudogene differed and/or that different rates of gene conversion operated on the pseudogene in the cpDNA of the three plant groups in Gramineae. The intergenic spacer regions diverged approximately ten times faster than the genes. The divergence of wheat from barley, and that from rice are estimated based on the nucleotide similarity to be 1.5, 10 and 40 million years, respectively.


Subject(s)
Chloroplasts , Genes, Plant , Triticum/genetics , Base Sequence , Biological Evolution , Molecular Sequence Data , Mutation , Oryza/genetics , Plant Proteins/genetics , Polymorphism, Genetic , Pseudogenes , Ribosomal Proteins/genetics , Sequence Alignment , Sequence Homology, Nucleic Acid
11.
Genetics ; 147(4): 1899-914, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9409845

ABSTRACT

To study the origin and maintenance mechanisms of the PGI allozyme polymorphism of a wild plant, Dioscorea tokoro, DNA sequences of the entire coding region (1701 bp) and two intronic regions (total 2049 bp) of the Pgi gene as well as a part of the Adh gene (590 bp) were analyzed. Two replacement substitutions were revealed to be responsible for the differentiation of three allozymes alleles (Pgi-a, Pgi-b and Pgi-c) that occur in natural population in intermediate frequencies. Interspecific comparison of DNA sequences identified Pgi-b as the oldest allele, from which two other alleles were derived probably within the last 150,000 years. The level of DNA polymorphism at D. tokoro Pgi locus was low. No elevated level of DNA polymorphism was detected in the close vicinity of the two replacement sites differentiating the three allozymes. Departures from the neutral mutation hypothesis were detected by Fu and Li's and MK tests. The observed patterns of DNA polymorphism are explainable by both (1) the neutral mutation hypothesis with an assumption of small effective size of D. tokoro population, and (2) the positive selection hypothesis that the allele frequencies of Pgi-a and Pgi-c have increased in a short time by their selective advantages.


Subject(s)
DNA, Plant , Glucose-6-Phosphate Isomerase/genetics , Polymorphism, Genetic , Solanaceae/enzymology , Alcohol Dehydrogenase/genetics , Amino Acid Sequence , Amino Acids/metabolism , Base Sequence , Genes, Plant , Introns , Molecular Sequence Data , Solanaceae/genetics , Solanaceae/metabolism
12.
Clin Cancer Res ; 6(8): 3166-71, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10955799

ABSTRACT

Few attempts have been made at the molecular detection of urothelial cancer cells in the blood or lymph nodes mainly because of an absence of good candidate molecular or genetic changes specific to urothelial cancer or urothelium. In 1990, however, genes that encode urothelium-specific transmembrane proteins, uroplakins (UPs), were cloned. We have established a method of detecting circulating cancer cells in peripheral blood of patients with transitional cell carcinoma by nested reverse transcription-PCR assay for UP II. UP II mRNA-positive cells were detected in 3 (10.3%) of 29 patients with superficial cancers (pTa-1N0M0), 4 (28.6%) of 14 patients with muscularly invasive cancers (pT2-4N0M0), 2 (40.0%) of 5 loco-regional node-positive patients (pN1-2M0), and 6 (75.0%) of 8 patients with distant metastases. Positive rates, therefore, increased with tumor extension (P = 0.0033, Kruskal-Wallis test). Furthermore, sequential blood sampling was performed in three patients with metastases during and after systemic chemotherapy, and UP-II-positive cells were found to have disappeared in two patients who responded well to the systemic chemotherapy. These results suggest that our nested reverse transcription-PCR assay for UP II is highly specific and might be used as a tumor marker for molecular staging of urothelial cancers, although the sensitivity is not so optimal.


Subject(s)
Biomarkers, Tumor/blood , Carcinoma, Transitional Cell/blood , Membrane Proteins/blood , Neoplastic Cells, Circulating/metabolism , Urologic Neoplasms/blood , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/biosynthesis , Biomarkers, Tumor/genetics , Carcinoma, Renal Cell/blood , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/metabolism , Carcinoma, Squamous Cell/blood , Carcinoma, Squamous Cell/metabolism , Carcinoma, Transitional Cell/genetics , Carcinoma, Transitional Cell/metabolism , Female , Humans , Kidney Neoplasms/blood , Kidney Neoplasms/genetics , Kidney Neoplasms/metabolism , Lymphatic Metastasis , Male , Membrane Proteins/biosynthesis , Membrane Proteins/genetics , Middle Aged , Neoplasm Metastasis , RNA, Messenger/biosynthesis , RNA, Messenger/blood , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Urinary Bladder Neoplasms/blood , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/metabolism , Urologic Neoplasms/genetics , Urologic Neoplasms/metabolism , Uroplakin II
13.
Int J Oncol ; 19(1): 19-24, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11408917

ABSTRACT

The caspase family of proteases is speculated to have a crucial role in apoptosis. The effect of treatment with adriamycin (ADR), cisplatin (CDDP), 5-fluorouracil (5-FU), vinblastine (VLB), IFN-alpha, or IFN-gamma on the activation of caspase-3, -6, -8, and -9 in renal cell carcinoma (RCC) cells was investigated, to clarify the mechanisms of chemo- and immunotherapeutic agent-mediated apoptosis. Caspase activity was determined by a quantitative colorimetric assay. Apoptosis was monitored by acridine-orange staining assay. Treatment of ACHN cells with CDDP, VLB, IFN-alpha, or IFN-gamma did not activate caspase-3, but its activity was increased 7.2-fold (p = 0.0001) with ADR and 2.8-fold (p = 0.0385) with 5-FU in comparison with control. Furthermore, when the ADR treatment time was shortened from 24 to 8 or 2 h, the same caspase-3 activation occurred. Activation of caspase-3 was also observed in six freshly isolated human RCC cells after the treatment with ADR. Of the six freshly derived RCC cells treated with 5-FU, caspase-3 activity was increased 3.1-fold (p = 0.0051) and 2.4-fold (p = 0.0346) in two of them, respectively. Epirubicin and pirarubicin, compounds closely related to ADR, also respectively enhanced 4.2-fold (p = 0.0052) and 2.8-fold (p = 0.0147) caspase-3 activity in ACHN cells. The activation of caspase-3 observed with a colorimetric assay was confirmed with immunocytochemical analysis using the anti-active caspase-3 mAb, which specifically recognizes the active form of caspase-3. Furthermore, both active caspase-3 and apoptosis triggered by either ADR or 5-FU were inhibited significantly by the general caspase inhibitor Z-VAD-FMK, or a specific caspase-3 inhibitor DMQD-CHO. These findings provide a mechanistic explanation for anthracyclines and 5-FU induced-apoptosis.


Subject(s)
Antibiotics, Antineoplastic/pharmacology , Carcinoma, Renal Cell/enzymology , Caspases/metabolism , Fluorouracil/pharmacology , Kidney Neoplasms/enzymology , Amino Acid Chloromethyl Ketones/pharmacology , Apoptosis/drug effects , Carcinoma, Renal Cell/pathology , Caspase 3 , Caspase Inhibitors , Cysteine Proteinase Inhibitors/pharmacology , Enzyme Activation/drug effects , Humans , Immunoenzyme Techniques , Interferon-alpha/pharmacology , Interferon-gamma/pharmacology , Kidney Neoplasms/pathology , Tumor Cells, Cultured/enzymology , Tumor Cells, Cultured/pathology
14.
J Mass Spectrom ; 34(8): 885-8, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10423570

ABSTRACT

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.


Subject(s)
DNA/chemistry , Polymorphism, Genetic , Receptors, Androgen/genetics , Trinucleotide Repeats , Base Sequence , DNA/blood , Humans , Leukocytes , Male , Mass Spectrometry/methods , Molecular Sequence Data , Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
15.
Urology ; 53(1): 53-9, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9886588

ABSTRACT

OBJECTIVES: Recent clinical trials have implied the cytotoxic and antiproliferative effects of combining 5-fluorouracil and interferon-alpha in the treatment of metastatic renal cell cancer. We therefore conducted an open multicenter trial to test the efficacy of such a combination on this cancer. METHODS: Human lymphoblastoid interferon (3 MIU per patient) was administered subcutaneously three times weekly for 12 weeks, while 5-fluorouracil was administered (600 mg/m2/day) as a continuous infusion for the first 5 days, followed by an intravenous bolus infusion of 600 mg/m2 once a week from the 3rd week until the 12th week. RESULTS: Of the 63 patients entered into the trial, 55 were eligible and evaluable for systemic toxicities, and 53 were evaluable for their response. All patients had undergone a prior nephrectomy, and their European Cooperative Oncology Group (ECOG) performance status ranged from 0 to 3 (median 0). Three complete and eight partial responses were induced, with an overall response rate of 20.0%. The median time to progression and the median survival time were 11 and 33 months, respectively. World Health Organization grade 3 toxicities were observed in 8 patients; however, no grade 4 toxicities or toxicity-related deaths were noted. CONCLUSIONS: Combination therapy of interferon-alpha plus 5-fluorouracil at the above-described dosage and schedule produced no better responses than interferon monotherapies. Prolongation of survival could be attributable to the fair performance status of the patients. This regimen has limited value for the treatment of patients with advanced renal cell cancer.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Carcinoma, Renal Cell/drug therapy , Carcinoma, Renal Cell/secondary , Kidney Neoplasms/pathology , Adult , Aged , Carcinoma, Renal Cell/mortality , Disease-Free Survival , Female , Fluorouracil/administration & dosage , Humans , Interferon-alpha/administration & dosage , Male , Middle Aged , Survival Rate
16.
Oncol Rep ; 4(1): 39-41, 1997.
Article in English | MEDLINE | ID: mdl-21590008

ABSTRACT

Inhalation of interferon-gamma (IFN-gamma) was used in four patients with pulmonary metastasis from renal cell carcinoma. Three million JRU IFN-gamma were inhaled every day (3 patients) or 5 times per week (1 patient). One patient achieved no change after IFN-gamma inhalation and three patients had no response. No drug-related side effects were observed. These findings point to the clinical feasibility of inhalation of IFN-gamma. However, further studies are needed to establish the anti-tumor efficacy of IFN-gamma inhalation therapy.

17.
Biomed Pharmacother ; 54 Suppl 1: 211s-214s, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10915027

ABSTRACT

A total of 370 laparoscopic adrenalectomies, including 311 transperitoneal (TP) and 59 retroperitoneal (RP) approaches, were performed in nine urologic centers, where the laparoscopic adrenalectomy was first begun independently in Japan, and their affiliated hospitals between January 1992 and September 1996. The clinical diagnoses of those 370 adrenal diseases were primary aldosteronism in 155 patients, Cushing's syndrome in 61. preclinical Cushing's syndrome in 21. pheochromocytoma in 16, nonfunctioning adenoma in 87, complicated cyst in ten, myelolipoma in nine, adrenal cancer in four and other diagnoses in eight (table 1). There was no mortality in this series. Intraoperative complication rate was 33/370 (9%) in total: 26/311(8%) in the TP procedures and 7/59 (12%) in the RP procedures (table 11). Postoperative complication rate was 24/370 (6%) in total: 22/311 (7%) in the TP procedures and 2/59 (3%) in the RP ones (table 111). Conversion rates to open surgery in total, in the TP and in the RP procedures were 13/370 (3.5%), 10/311 (3.2%) and 3/59 (5.1 %). respectively (table IV). Although the RP procedure has a lower morbidity rate compared to the TP procedure, more skill is required to overcome the drawback of the narrow working space and fewer anatomical landmarks.


Subject(s)
Adrenal Gland Neoplasms/surgery , Adrenalectomy/adverse effects , Laparoscopy/adverse effects , Adenoma/surgery , Cushing Syndrome/surgery , Humans , Hyperaldosteronism/etiology , Hyperaldosteronism/surgery , Japan , Peritoneal Cavity/anatomy & histology , Peritoneal Cavity/surgery , Pheochromocytoma/surgery , Retrospective Studies
18.
J Endourol ; 11(5): 361-5, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9355955

ABSTRACT

Between July 1992 and October 1996, 100 transperitoneal laparoscopic adrenalectomies were performed on 99 patients at our hospital and affiliated hospitals. The clinical diagnoses were primary aldosteronism (41 patients), Cushing's syndrome (15), pre-Cushing's syndrome (6), pheochromocytoma (7; 8 adrenal glands), adrenal cancer (2), nonfunctioning adenoma (22), myelolipoma (3), and complicated adrenal cyst (3). Ninety-seven glands were removed laparoscopically. The mean operative time was 240 +/- 76 (SD) minutes and the mean blood loss 68 +/- 80 mL for the series. The mean blood was 77 +/- 113 mL when the three operations that were converted to open surgery are included. The mean times for the return to a normal diet and unassisted ambulation were 1.3 +/- 0.6 and 1.4 +/- 0.8 days, respectively. The mean duration of the use of analgesics was 1.5 +/- 1.3 days, including the day of surgery. In contrast, in the latest 10 open adrenalectomies done at Kyoto University Hospital, the mean operative time was 186 +/- 53 minutes and the mean blood loss 220 +/- 170 mL. The mean times for return to a normal diet and for unassisted ambulation and the mean duration of the use of analgesics were 1.9 +/- 0.3, 2.9 +/- 1.1, and 2.9 +/- 1.7 days, respectively. Thirty-six operations, excluding one converted to open surgery, performed at Kyoto University Hospital were selected to look at the learning curve for transperitoneal laparoscopic adrenalectomy and evaluated for operative time and blood loss. The mean operative time and mean blood loss in the first 10 procedures performed at Kyoto University Hospital were 256 +/- 63 minutes and 89 +/- 57 mL; however, these values were reduced to 177 +/- 39 minutes and 48 +/- 32 mL in the next 10 procedures at the same hospital. Laparoscopic adrenalectomy via the transperitoneal anterior approach can be equivalent to open adrenalectomy in efficiency with a shorter convalescence.


Subject(s)
Adrenal Gland Diseases/surgery , Adrenalectomy/methods , Laparoscopy , Adult , Aged , Humans , Middle Aged
19.
Hinyokika Kiyo ; 41(9): 697-701, 1995 Sep.
Article in Japanese | MEDLINE | ID: mdl-7484536

ABSTRACT

Approximately 15% of cases of renal cell carcinoma (RCC) present cystic configuration on radiologic and pathologic examination. The mechanisms of cyst formation in RCC are intrinsic multilocular cystic growth (multiloculated renal cell carcinoma: MCRCC), unilocular cystic growth (cystadenocarcinoma), cystic necrosis and tumor growth from the epithelial lining of a preexisting cyst. These cystic lesions accompanying RCC are often difficult to differentiate from the multiloculated renal cyst (MLC) or other benign cystic lesions such as hemorrhagic cyst and so on. Differential diagnosis of the complicated renal cystic lesions is discussed in this review.


Subject(s)
Carcinoma, Renal Cell/diagnosis , Kidney Diseases, Cystic/diagnosis , Kidney Neoplasms/diagnosis , Carcinoma, Renal Cell/complications , Carcinoma, Renal Cell/therapy , Diagnosis, Differential , Humans , Kidney Diseases, Cystic/complications , Kidney Neoplasms/complications , Kidney Neoplasms/therapy , Magnetic Resonance Imaging , Tomography, X-Ray Computed
20.
Hinyokika Kiyo ; 33(7): 1096-9, 1987 Jul.
Article in Japanese | MEDLINE | ID: mdl-3687634

ABSTRACT

We report two cases of small renal cell carcinoma, which were incidentally detected by abdominal computed tomography (CT) and ultrasound. Both tumors were less than 15 mm in diameter and low grade, stage I lesions. Incidentally found renal cancer does not always indicate better prognosis. However, in selected patients with small tumors, surgical enucleation may be an acceptable method of treatment even if the contralateral kidney is normal. Renal screening should be a routine part of abdominal ultrasound. In our institution, ultrasound revealed incidental renal cancer in 0.035% of non-urologic patients.


Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Female , Humans , Male , Middle Aged
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