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Biomedical subjects

H Mano

Publications and source records attributed to H Mano.

At least 109 records · Page 6Linked to original sources
PubMed1993

Coordinate expression of the alpha and beta chains of human granulocyte-macrophage colony-stimulating factor receptor confers ligand-induced morphological transformation in mouse fibroblasts.

Two distinct components, alpha and beta chains, which compose the high affinity receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF) do not contain any catalytic domains of known enzymes. However, in mouse lymphoid cell lines transfected with cDNAs of the both chains, GM-CSF triggers tyrosine phosphorylation of several cellular proteins and allows continuous proliferation. To elucidate whether the high affinity receptor functions in nonhematopoietic cells, we have reconstituted human GM-CSF receptor in mouse NIH3T3 fibroblasts. In NIH3T3 clones, in which the high affinity receptor is reconstituted, human GM-CSF has triggered rapid tyrosine phosphorylation of cellular proteins, transfected beta chain, and another protein of 40-45 kDa. Moreover, human GM-CSF stimulated DNA synthesis and induced morphological transformation. These observations indicate that coordinately expressed alpha and beta chains of human GM-CSF receptor activates intrinsic protein-tyrosine kinases by the stimulation with human GM-CSF and that the activated protein-tyrosine kinases phosphorylate tyrosine residues of an intrinsic 40-45-kDa protein and the transfected beta chain in NIH3T3 cells. Activation of the protein-tyrosine kinases is likely to have biological functions to induce DNA synthesis and morphological transformation of mouse fibroblasts.

3T3 Cells↗
PubMed1993

Erythropoietin induces tyrosine phosphorylation and kinase activity of the c-fps/fes proto-oncogene product in human erythropoietin-responsive cells.

Erythropoietin (EPO) is a hematopoietic growth factor that stimulates the proliferation and differentiation of erythroid progenitor cells. Although the EPO receptor has no kinase domain, EPO rapidly induces tyrosine phosphorylation of several proteins in EPO-responsive cells. Therefore, the receptor activation by the ligand could induce tyrosine-kinase activity of unidentified cellular protein(s). Here we show that c-fps/fes proto-oncogene product (p92c-fes), nonreceptor tyrosine kinase, is tyrosine-phosphorylated on treatment with EPO in a human erythroleukemia cell line TF-1 that is responsive to granulocyte-macrophage colony-stimulating factor, interleukin-3, and EPO. In addition, the kinase activity of p92c-fes was shown to be enhanced by treatment with EPO. Therefore, p92c-fes could be implicated in a signaling pathway triggered by EPO in human EPO-responsive cells.

Amino Acid Sequence↗
PubMed1993

Mutations of the p53 gene in myelodysplastic syndrome (MDS) and MDS-derived leukemia.

The p53 gene is currently thought to be a tumor suppressor gene, and its alterations have been suggested to be involved in the pathogenesis of several human malignancies, including some leukemias and lymphomas. We present here evidence for the possible involvement of p53 gene mutations in the myelodysplastic syndrome (MDS), although the incidence is relatively low. Forty-four patients with MDS and six patients with overt leukemias that developed from MDS were studied for p53 gene alterations using reverse transcriptase-polymerase chain reaction, single-strand conformation polymorphism analysis, and nucleotide sequencing. Three patients with MDS (2 RAEB and 1 RAEB in T) had missense point mutations in the conserved regions of the p53 coding sequence. Furthermore, expression of the wild-type p53 mRNA was not detected in these three patients. The probable absence of normal p53 function in the three cases studied here suggests that alterations in the p53 gene may occasionally play a role in MDS. These three MDS patients with p53 gene mutations and an MDS-derived erythroleukemia cell line that we had previously reported to carry a p53 gene mutation showed no N-ras gene mutations, suggesting heterogeneity in the oncogenic mechanism of MDS.

Amino Acid Sequence↗
PubMed1993

Detection of human papillomaviruses from histologically normal lymph nodes of Japanese cervical cancer patients by nested polymerase chain-reaction assay.

To determine the prognostic significance of human papilloma virus (HPV) DNA in histologically normal lymph nodes, we developed a nested polymerase chain-reaction (PCR) method on HPV16,18 and 33 DNAs for formalin-fixed, paraffin-embedded tissues. We investigated 370 histologically normal lymph nodes from 15 patients treated for stage-IB/IIB (FIGO) invasive cervical cancer. HPV16 DNA was detected in 7 (47%) and HPV18 DNA in 3 (20%) of the cervical cancers. Examination of histologically normal lymph nodes from these 10 patients by nested PCR revealed HPV DNA in 5 (50%) of them; in all cases HPV type in lymph nodes and tumor was the same. Two of these 5 patients had a recurrence (pelvic cavity or lung) and died of cancer, although all 5 had had pelvic radiotherapy after radical hysterectomy and lymphadenectomy. These findings indicate that nested PCR is useful for evaluating early lymph-node involvement retrospectively in HPV-positive cases.

Adult↗
PubMed1993

Thyroid hormone affects the gene expression of retinoid X receptors in the adult rat.

Gene expression of three nuclear retinoid X receptors (RXR alpha, beta and gamma) was examined by Northern blot analysis in various rat tissues. The RXR alpha mRNA (5.5 kb) was detected in most tissues and particularly expressed at a high level in the liver. The RXR beta transcripts (2.4 and 3.0 kb) were expressed ubiquitously, and particularly at high levels in the brain and testis. In the liver, heart, kidney and lung, the RXR gamma mRNA (2.0 kb) was specifically detected. Furthermore, we examined the effect of retinoid, vitamin D and thyroid hormone status on the gene expression of RXR alpha, beta and gamma. Though retinoid and vitamin D did not affect the mRNA levels of three RXRs, the mRNA levels of two genes were controlled by thyroid hormone. Namely, positive (RXR beta) and negative (RXR gamma) regulations by thyroid hormone were observed with no effect on the gene expression of RXR alpha. These results suggest that thyroid hormone might affect the signal transduction of retinoid, vitamin D and thyroid hormone by changing RXR levels.

Animals↗
PubMed1993

c-fps/fes protein-tyrosine kinase is implicated in a signaling pathway triggered by granulocyte-macrophage colony-stimulating factor and interleukin-3.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) are hematopoietic growth factors which stimulate the proliferation and differentiation of myeloid progenitor cells. There is a considerable degree of overlap in target cell specificity and the functional effects of GM-CSF and IL-3. GM-CSF and IL-3 induce a nearly identical pattern of protein-tyrosine phosphorylation in certain cell lines, although their receptors have no kinase domains. Furthermore, their receptor complexes share one subunit (designated as beta). These observations raise the possibility that GM-CSF and IL-3 have a common signaling pathway. Here we show that both GM-CSF and IL-3 induce tyrosine phosphorylation and kinase activity of the c-fps/fes proto-oncogene product (p92c-fes), a non-receptor protein-tyrosine kinase, in a human erythro-leukemia cell line, TF-1, which requires GM-CSF or IL-3 for growth. In addition, GM-CSF induces physical association between p92c-fes and the beta chain of the GM-CSF receptor. p92c-fes is therefore a possible signal transducer of several hematopoietic growth factors including GM-CSF and IL-3 through the common beta chain.

Amino Acid Sequence↗
PubMed1993

Tumoricidal effect of human macrophage-colony-stimulating factor against human-ovarian-carcinoma-bearing athymic mice and its therapeutic effect when combined with cisplatin.

The effect of human macrophage-colony-stimulating factor (hM-CSF) on tumoricidal activity was examined in athymic mice bearing the human ovarian cancer cell line, HRA, injected intraperitoneally (i.p.). The survival period and survival rate in the groups treated daily with hM-CSF were significantly longer (P < 0.01) than in the untreated group. The peritoneal cell smears showed that ascitic tumor cells were markedly decreased in the hM-CSF-treated groups, and macrophages phagocytosed tumor cells, indicating a contact-mediated direct cytolysis. The combined therapeutic effects of cisplatin and hM-CSF on HRA-bearing athymic mice were also studied. The mean survival period was 25.4, 47.2, 42.4 and 67.4 days, respectively, in the untreated group, and in the groups treated with cisplatin alone, with hM-CSF alone, and with combined cisplatin and hM-CSF. The survival period and rate were significantly longer (P < 0.01) in the group treated with combined cisplatin and hM-CSF than in those treated with cisplatin or hM-CSF alone, indicating the therapeutic effectiveness of the combined use. Moreover, hM-CSF is effective against granulocytopenia due to bone marrow suppression caused by cisplatin. Our data demonstrate that hM-CSF administered i.p. has a tumoricidal activity in athymic mice bearing human ovarian cancer i.p., which is mediated by activated macrophages, and that the combined administration of cisplatin and hM-CSF has a significant therapeutic effect.

Agranulocytosis↗
PubMed1993

Mutagenicity of blue rayon extracts of human bile in the Ames test.

The mutagenicity of human bile was examined in the Ames Salmonella/microsome assay. Bile samples were obtained from the gallbladders resected from patients with cholelithiasis, choledocholithiasis, gallbladder cancer, extrahepatic bile duct cancer and other diseases. For extraction of mutagenic components, the bile samples were treated with blue rayon and the adsorbed materials were assayed with Salmonella typhimurium TA98 in the presence of S9 mix. Twenty-four bile samples were tested and positive mutagenic activity was found in 14 samples. A 200-microliter bile equivalent material gave 6.3 times as many revertant colonies as the solvent control. With several samples that had undergone two cycles of blue rayon extraction, clear dose-response relationships in mutagenicity were demonstrated.

Adult↗
PubMed1993

Detection of the PML/RAR alpha fusion gene in acute promyelocytic leukemia with a complex translocation involving chromosomes 15, 17, and 18.

We report a case with typical clinical features of acute promyelocytic leukemia (APL) carrying an atypical chromosomal aberration involving chromosomes 15, 17, and 18. Molecular analysis using Southern blot hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR) proved the creation of the PML/RAR alpha fusion gene in this case. These findings support the notion that this fusion is of crucial importance to leukemogenesis of APL.

Base Sequence↗
PubMed1993

Mutagenicity of activated carbon adsorbate of drinking water in the Ames assay.

Mutagenicity of activated carbon adsorbate from drinking water collected in Niigata City was assayed by the Ames assay. Adsorbate was extracted from activated carbon with benzene, and then with ethanol. Although the benzene extract was not mutagenic, the ethanol one showed the mutagenic activity for Salmonella typhimurium strains TA98 and TA100 with and without S9 mix. The ethanol extract was much more mutagenic on TA100 than TA98 both with and without S9 mix. The mutagenic activity per liter of water was found to be the strongest in winter and the weakest in summer.

Animals↗
PubMed1993

Gene expression of cellular retinol-binding protein I (CRBP I) is affected by dietary proteins in the rat liver.

The effect of dietary proteins and vitamin A status on the gene expression of cellular retinol-binding protein I (CRBP I) was studied in the rat liver. The gene expression was estimated as amounts of transcript (mRNA) by Northern blot analysis using rat CRBP I cDNA. Though vitamin A status is known to positively regulate the gene expression of CRBP I in the extrahepatic tissues, in the present study we observed that the amount of the CRBP I transcript in liver was neither reduced by vitamin A-deficiency, nor affected by replenishment with an excess dose of all-trans retinoic acid. These results indicate that in the liver, different from the extrahepatic tissues, the gene expression of CRBP I may not be controlled by vitamin A. However, when the rats were fed on the diets that differed in dietary proteins, the gene expression of CRBP I in liver was enhanced by higher quality and quantity of dietary proteins, though no effect of dietary proteins was observed upon the hepatic contents of retinol. The concentrations of serum retinol were almost proportional to the mRNA levels of CRBP I. In contrast, the hepatic gene expression of another retinol-binding protein, RBP, and one subtype of retinoic acid receptor, RAR alpha was not influenced in the nutritional condition tested here. Our findings suggest that the gene expression of CRBP I in liver may be under control of the intake of dietary proteins. Thus, it is likely that in the light of the function of CRBP I on cellular transport and metabolism of retinol, dietary proteins may affect the actions of vitamin A in the extrahepatic tissues through changing the amounts of CRBP I in liver.

Animals↗
PubMed1993

Expression of a novel form of Tec kinase in hematopoietic cells and mapping of the gene to chromosome 5 near Kit.

The Tec kinase was initially identified as a novel cytoplasmic protein tyrosine kinase that is preferentially expressed in the liver and is highly homologous to the Drosophila Dsrc28C src-related tyrosine kinase. In screening of interleukin 3 (IL-3)-dependent myeloid leukemia cells for protein tyrosine kinases, we observed that all cell lines examined expressed high levels of Tec transcripts. However, characterization of Tec cDNAs indicated that they differed significantly from the published sequence. Most strikingly, an insertion of 41 bp in the 5' region affects the initiation codon and results in replacing the published 13 amino acid amino-terminal sequences with 94 amino acids. Using polymerase chain reaction (PCR) analysis, only the form containing the insertion was detected in hematopoietic cells. In addition, we found an in-frame insertion of 66 bp that introduces an additional 22 amino acids into the SH3 domain. This insertion restores conserved SH3 sequences that are found in the src gene family and in the Dsrc28C gene. By PCR analysis, approximately equal levels of Tec transcripts containing the intact SH3 domain and containing the 22 amino acid deletion were found in hematopoietic cells. Lastly, by interspecies backcross analysis, we show that the Tec gene is tightly linked to the c-Kit gene on mouse chromosome 5.

Amino Acid Sequence↗
PubMed1993

[Mutagen extraction from bile of patients with inflammatory biliary pathology: Ames test using blue rayon].

Gallbladder carcinoma is frequent in Chile. The aim of this study was to report the mutagenicity of whole human bile, using the Ames/Salmonella microsome assay with Salmonella typhimurium TA98. The bile of 19 patients, aged 23 to 64 years old, subjected to cholecystectomy was examined, and mutagen activity was found in 13 (72%). Mutagens were extracted using blue rayon and three dilutions for the eluted material from blue rayon were used (50, 100 and 200 ul). The best result was obtained using 200 ul. In some cases, the amount of revertive colonies was very high (over 5 times the control value). We propose that the bile from these patients possibly contains mutagenic substances with frame shift mutagenic activity and that these substances may be related to gallbladder carcinoma. Our results have addressed the importance of bile studies to elucidate the pathogenesis of gallbladder carcinoma.

Adult↗
PubMed1992

Effect of retinoid status on alpha, beta and gamma retinoic acid receptor mRNA levels in various rat tissues.

We have investigated the effects of retinoids, vitamin D and thyroid hormone on the levels of retinoic acid receptor (RAR)alpha, RAR beta and RAR gamma mRNAs in intact animals. Although vitamin A deficiency caused no significant changes in the levels of RAR alpha and RAR gamma mRNAs, the level of RAR beta transcripts was greatly decreased in various tissues of vitamin A-deficient rats, but was restored rapidly to a normal level after administration of retinoic acid. Retinol also restored the RAR beta mRNA level, but the magnitude and kinetics of the induction differed from those by retinoic acid. The use of specific inhibitors demonstrated that this autoregulation of RAR beta gene expression in vivo occurred at the transcriptional level. In addition, from these results it was postulated that the maintenance of the normal RAR beta mRNA levels seemed to require a threshold serum retinol concentration (about 25 micrograms/dl). Moreover, we found that administration of retinol and retinoic acid to normal rats caused the overexpression of RAR beta transcripts (2-15-fold) when compared with the control levels of RAR beta mRNA, although the levels of RAR alpha and RAR gamma mRNAs were not affected. Vitamin D and thyroid hormone did not modulate the levels of RAR transcripts. These findings clearly indicate the specific ligand regulation of RAR beta gene expression in intact animals. The altered levels of RAR beta according to retinoid status may affect retinoid-inducible gene expression.

Animals↗
PubMed1992

Identification of a soluble GM-CSF binding protein in the supernatant of a human choriocarcinoma cell line.

We identified two forms of the receptor for granulocyte-macrophage colony-stimulating factor (GM-CSF) made by the human choriocarcinoma cell line JEG-3 using an affinity-labeling technique. The protein was identified in the detergent-extract was 78 kDa, very similar to that of the membrane-bound GM-CSF receptor alpha chain expressed in a wide variety of hematopoietic and nonhematopoietic cells, including JEG-3. In contrast, a 62-kDa GM-CSF binding protein, or the soluble GM-CSF receptor, was identified in the supernatant of JEG-3 cells. Utilizing the same affinity labeling technique, we did not detect the soluble GM-CSF binding protein in the supernatant of several hematopoietic cell lines, such as U-937 and KG-1, which express membrane bound alpha chain as well as beta chain. The 62-kDa soluble GM-CSF receptor is produced in abundant amounts by JEG-3, but in very small amounts, if any, by hematopoietic cell lines.

Choriocarcinoma↗
PubMed1992

Prophylactic effect of bestatin on the onset of invasive mole--clinical and fundamental studies.

This study was performed to determine whether bestatin (Ubenimex) has clinical prophylactic effects on the onset of invasive mole and a direct inhibitory effect on the growth of hydatidiform molar cells. A total of 49 patients with hydatidiform mole treated at Nagoya University Hospital from 1984 to 1990 were randomly divided into two groups, a bestatin administered-group and a bestatin non-administered group. Patients in the bestatin group were given 30 mg of bestatin orally and daily for three months just after their molar deliveries. There was no significant difference in age, gravidity, parity and gestational weeks between the two groups. There was also no significant difference in the duration of human chorionic gonadotropin (hCG) negative conversion in patients without invasive mole between the two groups. However, the incidence of invasive mole in the bestatin group (2/25, 8%) was significantly lower than that of the non-bestatin group (7/24, 29.2%). Nevertheless, there was no significant difference between the two groups in such immunological parameters as PHA skin test, PPD skin test, PHA stimulation index (PHA-SI), white blood cell (WBC) count lymphocytes % per WBC, OKT 3% per lymphocytes, OKT 4% per lymphocytes, OKT4/OKT8 and Leu 11% per lymphocytes. In vitro studies were performed with primary cultured hydatidiform moles. The result was that bestatin inhibited the secretion of hCG and 3H-thymidine uptake of hydatidiform molar cells. Thus, a possibility was suggested that bestatin directly inhibits the growth of hydatidiform molar cells and prevents the onset of invasive mole.

Adolescent↗
PubMed1992

Acute toxicity of tri-n-butyltin chloride (TBTC) in the Syrian golden hamster.

The effects of tri-n-butyltin chloride (TBTC) on Syrian golden hamsters were studied. TBTC in single doses of 0, 29.6, 44.4, 66.7, 100 or 150 mg/kg were given to 10 male hamsters in each group and the mortality rates were determined two weeks thereafter. They were 0% (0/10), 0% (0/10), 10% (1/10), 10% (1/10), 30% (3/10) and 70% (7/10) for 0, 29.6, 44.4, 66.7, 100 and 150 mg/kg groups, respectively. In the case of females, seven groups consisting of 10 animals each were given TBTC at doses of 0, 29.6, 44.4, 66.7, 100, 150, or 225 mg/kg. The mortality rates determined two weeks after the TBTC treatment were 0% (0/10), 10% (1/10), 10% (1/10), 0% (0/10), 40% (4/10), 30% (3/10) and 90% (9/10) for 0, 29.6, 44.4, 66.7, 100, and 225 mg/kg groups, respectively. Based on these mortality data, the LD50s via oral administration were determined as 146.9 mg/kg (95% C.I. 111.8-193.3 mg/kg) for the male and 172 mg/kg (95% C.I. 127.2-233.4 mg/kg) for the female. Regarding pathological changes, animals experienced lesions in the bile duct, such as the dilatation of the common bile duct and cholestasis, and/or adhesion of the bile duct to the liver, gallbladder, pancreas and duodenum. In a separate experiment, a single dose of 44.4 mg/kg TBTC was administered orally to 12 male hamsters, then the concentrations of TBTC and its metabolite, di-n-butyltin chloride (DBTC), in the liver were analyzed by means of gas chromatography for 14 days after the treatment. The maximum concentrations of TBTC and DBTC appeared one day after the administration, and decreased rapidly thereafter. The concentration of DBTC was found to be higher than that of TBTC throughout the experimental period.

Alanine Transaminase↗