PubMed Health⌕ Search

Biomedical subjects

K Fukuchi

Publications and source records attributed to K Fukuchi.

At least 145 records · Page 8Linked to original sources
PubMed1990

[In vitro thermosensitivity of various human tumors evaluated using the SDI (succinate dehydrogenase inhibition) test].

In vitro thermosensitivity of various human tumors including 90 esophageal, 10 gastric and 40 colo-rectal cancers were evaluated using the succinate dehydrogenase inhibition (SDI) test. Tumor fragments minced with scissors were incubated at 43 degrees C as heat treated cells and at 37 degrees C as controls for 20 hrs, and assayed for the succinate dehydrogenase (SD) activity using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) as a hydrogen acceptor. The thermosensitivity was estimated by the percentage of SD activity of heat treated cells compared to that of each control. A variation in the thermosensitivity was noted between patients. The SD activity was 60.1 +/- 20.3% (mean +/- standard deviation) for esophageal cancers, 34.9 +/- 21.7% for gastric cancers, 50.3 +/- 20.6% for colo-rectal cancers. Significant differences were noted between esophageal cancers and gastric cancers, colo-rectal cancers (p less than 0.01 and p less than 0.05, respectively). When the thermosensitivity was arbitrarily defined as reduction in the SD activity to 50% of control or less, the positive rates were 31.1% for esophageal cancer, 70% for gastric cancer and 62.5% for colo-rectal cancer. Our results show that the SDI test is a useful method for determination of the thermosensitivity of clinical samples.

Digestive System Neoplasms↗
PubMed1990

[The sensitivity of 1,000 human tumors to antitumor drugs using the succinate dehydrogenase inhibition (SDI) test].

The chemosensitivity was evaluated by the in vitro succinate dehydrogenase inhibition (SDI) test in 1,000 human tumors including 237 gastric cancers, 116 colorectal cancers, 113 hepatoma and 534 others. These tumor cells were exposed to 5 kinds of antitumor drugs, carboquone (CQ), adriamycin (ADM), mitomycin C (MMC), aclacinomycin A (ACR), cis-platinum (DDP). After exposure to the antitumor drugs, cell viability was assessed with colorimetric assay, based on the ability of succinate dehydrogenase (SD) in living tumor cells to reduced a tetrazolium (MTT) to a formazan. The chemosensitivity was determined to be positive when the SD activity of drug exposed cells decreased to below 50% of that of control cells, on day 3 of exposure. The chemosensitivity varied in the tumor tissues. The chemosensitivity of metastatic lesions of lymph nodes were higher than that of the primary lesions, while metastatic liver tumors had lower sensitivity than the primary lesions. The intra-tumorous distribution of SD activity in 12 human gastric cancers were compared with normal adjacent tissues using histochemistry. Seventy-five % (9/12) of gastric cancer tissues had higher SD activity than normal adjacent tissues. The SDI test is rapid and simple method to predict the sensitivity test of various human tumors to antitumor drugs.

Antineoplastic Agents↗
PubMed1989

Mutator phenotype of Werner syndrome is characterized by extensive deletions.

Werner syndrome (WS) is a rare autosomal-recessive disorder characterized by the premature appearance of features of normal aging in young adults. The extensive phenotypic overlap between WS and normal aging suggests they may also share pathogenetic mechanisms. We reported previously that somatic cells from WS patients demonstrate a propensity to develop chromosomal aberrations, including translocations, inversions, and deletions, and that WS cell lines demonstrate a high spontaneous mutation rate to 6-thioguanine resistance. We report here the biochemical and molecular characterization of spontaneous mutations at the X chromosome-linked hypoxanthine phosphoribosyltransferase (HPRT) locus in 6-thioguanine-resistant WS and control cells. Blot hybridization analysis of 89 independent spontaneous HPRT mutations in WS and control mutants lacking HPRT activity revealed an unusually high proportion of HPRT deletions in WS as compared with control cells (76% vs. 39%). Approximately half (58%) of the deletions in WS cells consisted of the loss of greater than 20 kilobases of DNA from the HPRT gene. These results suggest that an elevated somatic mutation rate, and particularly deletions, may play pathogenetically important roles in WS and in several associated age-dependent human disease processes.

Adolescent↗
PubMed1988

A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys.

Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10 micrograms/ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10(-4) to 10(-5). A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell-free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell-free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanthine phosphoribosyltransferase expression may occur in dog and human kidney cells.

Animals↗
PubMed1988

The structure of Marek's disease virus DNA: amplification of repeat sequence in IRs and TRs.

The structure of Marek's disease virus (MDV) DNA was investigated by using Southern blot hybridization analysis. A heterogeneous region was observed in the inverted repeats region, IRs and TRs, as well as in the TRL and IRL. The results of DNA sequencing of the heterogeneous region showed that the heterogeneity of IRS and TRS was due to amplification of a 178-bp repeat sequence. Amplification of IRS and TRS was found in viral DNA from both pathogenic and nonpathogenic strains. The structure of DNA from the latent MDV genome present in established lymphoblastic cells was also determined. Amplification of the 132-bp repeat sequence in IRL and TRL was not found in latent MDV DNA of established lymphoblastic cells, whereas amplification of the 178-bp repeat sequence in IRS and TRS was found in the same DNA.

Base Sequence↗
PubMed1988

Role of medullasin in granulocytes in biophylaxis. Elastinolytic activity and the potentiation of cytostatic activity of human monocytes.

Medullasin, a serine protease in bone marrow cells, resembles elastase, but is essentially devoid of elastinolytic activity. The protease revealed elastinolytic activity when small amounts of other proteases such as trypsin, papain, chymotrypsin, or collagenase coexisted in the incubation mixture. In vitro treatment of human monocytes with medullasin caused an increment of their cytostatic activity. Since medullasin failed to increase the cytostatic activity in the supernatant of monocytes, the enhancement of cytostatic activity of monocytes by medullasin is considered to be not mediated through the production of soluble factors from monocytes.

Animals↗
PubMed1988

[Human colorectal carcinoma is more sensitive to HCFU than 5-FU and tegafur in in vitro and in vivo drug sensitivity tests].

The sensitivity of human colorectal cancer to 5-fluorouracil (5-FU) and its derivatives: 1-(tetrahydro-2-furyl)-5-FU (tegafur) and 1-hexylcarbamoyl-5-FU (HCFU) was determined by in vitro succinate dehydrogenase inhibition (SDI) test and in vivo subrenal capsule (SRC) assay. Using the SDI test with 25 colorectal cancer specimens, the succinate dehydrogenase (SD) activity decreased to 73.6 +/- 17.8% for 5-FU-treated cells and 37.2 +/- 17.0% for HCFU-treated cells, compared to that of control cells. The chemosensitivity-positive rates were 16% for 5-FU and 68% for HCFU. Using the SRC assay with 7 colorectal cancer specimens, the relative variation of tumor size, which was calculated by delta TS/TS0 = (TS6-TS0)/TS0 X 100 (%), decreased to -12.6 +/- 10.1% for 5-FU, -14.9 +/- 12.4% for tegafur and -23.9 +/- 14.2% for HCFU, and the inhibition of tumor growth following exposure to HCFU was evident. The chemosensitivity-positive rates were 49% for 5-FU, 57% for tegafur and 71% for HCFU. Our results show that HCFU is more effective to colorectal cancer than 5-FU and FT-207, and the chemosensitivity test of HCFU will be useful in determining the effective drug for the treatment of colorectal cancer.

Animals↗
PubMed1987

Analysis of steroid 21-hydroxylase gene in five unrelated Japanese patients with 21-hydroxylase deficiency.

DNA samples from five unrelated Japanese patients with 21-hydroxylase (21-OHase) deficiency were studied by Southern analysis using human 21-OHase cDNA. Patterns seen after digestion with not only TaqI but also KpnI showed that two out of the five patients were homozygous for a deletion of the 21-OHase B gene. This result supports the report that the 21-OHase B gene is functional. In the other three, smaller mutations might be responsible for the disorder. The parents of one of the two patients with the deletion had a common ancestor. Hybridization patterns of DNA from members of the family of the patient were consistent with an autosomal recessive mode of inheritance of the deletion that correlates with the clinical phenotype. The deletion segregated with HLA-Aw 24; Bw 61; Cw 3. Heterozygous carriers of 21-OHase deficiency could be detected by comparing the patterns as well as the HLA haplotypes in this family. The application of the family study to the prenatal diagnosis is also discussed.

Adrenal Hyperplasia, Congenital↗
PubMed1987

Medullasin enhances human natural killer cell activity by a mechanism other than the induction of interferons or interleukin-2.

The mechanism of activation of human natural killer cell activity by medullasin was studied with special reference to interferons and interleukin-2. Supernatant of lymphocytes treated with medullasin failed to stimulate natural killer cell activity when added to the lymphocyte suspension. Medullasin has been shown to enhance the natural cytotoxicity of lymphocytes stimulated with interferon-alpha or -beta. In the present study, medullasin increased the natural killer cell activity of lymphocytes stimulated with interferon-gamma. Treatment of lymphocytes with medullasin in the presence of antibodies to interferons (alpha, beta and gamma) did not alter the effect of medullasin. Medullasin also increased the natural cytotoxicity of lymphocytes stimulated with interleukin-2. The antibody to interleukin-2 failed to affect the increase of natural killer cell activity by medullasin. These results indicate that the mechanism of activation of human natural killer cell activity by medullasin is not mediated by the induction of interferons or interleukin-2.

Cells, Cultured↗
PubMed1987

[Comparison between the succinate dehydrogenase inhibition test and ATP assay for in vitro chemosensitivity testing].

The predicted level of cell viability was compared between the succinate dehydrogenase inhibition (SDI) test and the adenosine triphosphate (ATP) assay, both of which are used for in vitro human tumor chemosensitivity testing. After HeLa cells had been exposed to various concentrations of 5-FU for 1, 2, 3 or 4 days, the decrease occurring in viable cell number correlated with that of succinate dehydrogenase (EC 1.3.99.1) activity and that of the intracellular ATP level of the viable cells. In dead cells, the ATP level was extensively decreased, but the succinate dehydrogenase activity remained at a level of 11% of that of 5-FU-untreated viable cells, even on day 4. The cell viability correlated well with the intracellular ATP level, as compared with the succinate dehydrogenase activity. The activity remaining in dead cells must thus be taken into consideration for the prediction of chemosensitivity in the SDI test, but not in the ATP assay.

Adenosine Triphosphate↗
PubMed1986

Hutchinson-Gilford progeria syndrome in a 45-year-old man.

A 45-year-old man with typical Hutchinson-Gilford progeria syndrome is described. The patient had the characteristic physical findings of this syndrome, such as short stature, "horse-riding" stance, coxa valga, alopecia, micrognathia, craniofacial disproportion, and prominent eyes. He had refractory congestive heart failure due to arteriosclerotic heart disease and hypertension, and he also had arteriosclerosis obliterans. Some immunologic and endocrinologic abnormalities commonly seen in the elderly were present in this patient. On the basis of a review of the literature, this is the first patient with this syndrome who had survived into the fourth decade.

Arteriosclerosis Obliterans↗
PubMed1986

[In vitro chemosensitivity testing evaluated by intracellular ATP level: ATP assay].

In vitro chemosensitivity was evaluated by bioluminescence ATP assay in 12 human tumors including 7 gastric cancers and 5 colo-rectal cancers. Tumor fragments minced with scissors were exposed to 6 kinds of antitumor drugs: carboquone, adriamycin, aclacinomycin A, mitomycin C, cisplatin and 5-FU at peak plasma concentration or ten times peak plasma concentration. After 3 days at 37 degrees C, each tumor fragment suspension was washed with phosphate-buffered saline, boiled for 3 min and assayed for its intracellular ATP level using the luciferin-luciferase method. The ATP level of the drug-untreated group was 6.01 +/- 4.55 X 10(-10) moles/mg tissue protein in gastric cancers and 9.77 +/- 8.46 X 10(-10) moles/mg tissue protein in colo-rectal cancers. The percentages of cases in which the ATP level was reduced to less than 50% by the antitumor drugs were 70% for carboquone, 11% for adriamycin, 30% for aclacinomycin A, 45% for mitomycin C, 13% for cisplatin and 18% for 5-FU at peak plasma concentration, and 90% for carboquone, 78% for adriamycin, 80% for aclacinomycin A, 82% for mitomycin C, 63% for cisplatin and 82% for 5-FU at ten times peak plasma concentration. The test for chemosensitivity prediction is thus available at peak plasma concentration of antitumor drugs in ATP assay.

Adenosine Triphosphate↗
PubMed1986

[In vitro chemosensitivity of various human tumors evaluated by the succinate dehydrogenase inhibition (SDI) test (2)].

In vitro chemosensitivity was evaluated by succinate dehydrogenase inhibition (SDI) test in 94 human tumors including 59 gastric cancers, 27 colo-rectal cancers and 8 malignant lymphomas. Tumor fragments were exposed to 12 kinds of antitumor drugs at ten times peak plasma concentration. Evaluable rates were 86/94 (91%) for all cases, 56/59 (95%) for gastric cancers, 22/27 (81%) for colo-rectal cancers and 8/8 (100%) for malignant lymphomas. The mean of SD activity was decreased to 48% of that of control cells with aclacinomycin, 49% with carboquone, 53% with actinomycin D, 54% with mitomycin C and 54% with daunomycin for gastric cancers, 59% with adriamycin for colo-rectal cancers and 33% with cyclophosphamide (40487 S), and 33% with actinomycin D, 37% with vinblastine and 39% with adriamycin for malignant lymphomas. When the SD activity was reduced to below 50% by antitumor drugs, the chemosensitivity was defined as positive. The antitumor drugs which had a higher chemosensitive-positive rate were aclacinomycin, carboquone and mitomycin C for gastric cancers, adriamycin for colo-rectal cancers and 40487 S, daunomycin and vinblastine for malignant lymphomas. Our results suggest that the origin of a tumor is a critical factor in its chemosensitivity.

Antineoplastic Agents↗
PubMed1985

Cell fusion studies in the Werner syndrome.

The mechanism of retarded DNA synthesis was investigated by the cell fusion method. The [3H] TdR labeling index of young cells from patients with Werner syndrome (WS cells) X young normal human diploid fibroblast cells (NH cells) was considerably lower than that of NH homodikaryons, but was significantly higher than that of WS homodikaryons. The labeling index of old WS X young NH heterodikaryons was as low as that of WS homodikaryons. The labeling index of WS X HeLa heterodikaryons was the same as that of HeLa homodikaryons. These results indicate that WS cells are somewhat similar to senescent NH cells. The labeling indices in both WS karyoplast X NH heterodikaryons and WS cytoplast X NH cybrids were lower than those in normal karyoplast X NH heterodikaryons and normal cytoplast X NH cybrids, respectively. These results indicate that both nuclear and cytoplasmic environments are involved in the retarded DNA synthesis in WS cells. The most plausible interpretation of all our data is that the retarded DNA synthesis in WS cells could be caused by either the 'senescent factor(s)' or the deficiency of gene product(s) that is necessary for DNA synthesis and could be a secondary consequence of the genetic defect.

Adult↗