Molecular-dynamics simulations of SiH3 radical deposition on hydrogen-terminated silicon (100) surfaces.
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Journal Article↗
Biomedical subjects
T Ohira
Publications and source records attributed to T Ohira.
Long-term human hematopoiesis in SCID-hu mice bearing transplanted fragments of adult bone and bone marrow cells.
An attempt was made to establish an SCID-hu murine model of long-term human hematopoiesis by coimplantation of a bone fragment and bone marrow (BM) cells from an adult human. The SCID-hu mice were treated with a cytokine mixture (recombinant human stem cell factor, interleukin-3, granulocyte/macrophage colony-stimulating factor, and granulocyte colony-stimulating factor) for 4 months and were then maintained for further 8 months under cytokine-free conditions. In the peripheral blood, spleen, and implanted bone fragments in the SCID-hu mice that had received both a bone fragment and BM cells, human CD59+ cells were detected 1 year after transplantation; however, they were not detectable in SCID-hu mice that had received either a bone fragment or BM cells only. Thus, implantation of both a bone fragment and BM cells appears to provide a model of long-term adult human hematopoiesis in SCID mice.
Aged↗
In vitro cytotoxicity of a novel antitumor antibiotic, spicamycin derivative, in human lung cancer cell lines.
Spicamycin (SPM), produced by Streptomyces alanosinicus, induces potent differentiation in a human leukemia cell line, HL60. One of the derivatives of SPM (SPM-D), KRN5500, has a wide range of antitumor activity against human cancer cell lines. We examined the cytotoxicity of SPM-D in small and non-small cell lung cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony assays. SPM-D was active against a wide range of lung cancer cell lines. All three cisplatin (CDDP)-resistant cell lines established in our laboratory (PC-9/CDDP, PC-14/CDDP, and H69/CDDP) showed collateral sensitivity to SPM-D with relative resistance values of 0.43, 0.34, and 0.32, respectively. Intracellular SPM-D in PC-14/CDDP was 35% higher than that for PC-14 suggesting that intracellular accumulation can explain the collateral sensitivity to SPM-D at least in PC-14/CDDP. On the other hand, in PC-9/CDDP cells, no increase of intracellular SPM-D accumulation was observed, but the conversion ratio of a metabolite (the amino nucleoside moiety of spicamycin binding with glycine, SAN-G) from SPM-D evaluated by TLC was higher as compared with that of parental PC-9 cells (45.5% versus 37%; PC-9/CDDP versus PC-9). The increased intracellular metabolism of SPM-D could explain the mechanism of collateral sensitivity in PC-9/CDDP cisplatin-resistant cell lines. To elucidate the determinant of the SPM-D-induced cytotoxicity, we established SPM-D-resistant cell lines, PC-9/SPM-D, PC-14/SPM-D, and H69/SPM-D, by exposing cells to stepwise increases in SPM-D concentration. The relative resistances of these sublines were more than 5000, 46.6, and 37.8 times those of the parental cell lines, respectively. The intracellular concentration of the active metabolite, SAN-G, was found to be decreased in the SPM-D-resistant sublines. This result indicates that the intracellular metabolism of SPM-D to SAN-G is one of the determinants of cellular sensitivity to SPM-D in these SPM-D-resistant cell lines. In conclusion, both drug accumulation and metabolism may contribute to the sensitivity/resistance to SPM-D and both may merit investigation.
Adenocarcinoma↗
In vivo screening models of cisplatin-resistant human lung cancer cell lines using SCID mice.
In vivo screening models of a cisplatin (CDDP)-resistant human small-cell lung cancer cell (SCLC) line, H69/CDDP, and a non-small-cell lung cancer cell (NSCLC) line, PC-14/CDDP, were evaluated. The transplantability of the tumor xenografts to SCID mice was more than 90%. Tumor xenografts of H69/CDDP and PC-14/CDDP showed CDDP resistance during in vivo treatment. The novel anticancer agent 254-S showed only a partial effect on the growth of H69/CDDP and PC-14/CDDP while ormaplatin showed no cross resistance to CDDP. The in vivo results correlated well with the results of the in vitro MTT assay. In this in vivo sensitivity test, H69/CDDP and PC-14/CDDP were more sensitive to ormaplatin than its parental cell lines. In vivo sensitivity testing using SCID mice bearing transplanted CDDP-resistant tumors was shown to be useful for evaluating the effects of new anti-cancer drugs, especially those that might overcome CDDP resistance.
Animals↗
Intracellular carboxyl esterase activity is a determinant of cellular sensitivity to the antineoplastic agent KW-2189 in cell lines resistant to cisplatin and CPT-11.
KW-2189, a novel antitumor antibiotic belonging to the duocarmycins, possesses marked DNA-binding activity upon activation by carboxyl esterase to its active form, DU-86. Three duocarmycins, KW-2189, DU-86 and duocarmycin SA, were active against the cisplatin (CDDP)-resistant human non-small cell lung cancer cell lines PC-9/CDDP and PC-14/CDDP, and the multidrug-resistant human small cell lung cancer cell line H69/VP. However, HAC2/0.1, a CDDP-resistant human ovarian cancer cell line which is also resistant to CPT-11 because of decreased intracellular activation of CPT-11, was about 12.8-fold more resistant to KW-2189. HAC2/0.1 was not resistant to other duocarmycins as compared to its parental cell line, HAC2. There was no difference between HAC2 and HAC2/0.1 with regard to the intracellular accumulation of KW-2189. Addition of 130 mU/ml of carboxyl esterase to the culture medium did not influence the sensitivity of HAC2 cells to KW-2189. However, the sensitivity of HAC2/0.1 cells to KW-2189 was enhanced to the level of HAC2. These results suggest that HAC2/0.1 is less potent than HAC2 in activating KW-2189. The carboxyl esterase activity of whole-cell and microsomal extracts from HAC2/0.1 was approximately 60% of that from HAC2. The cell-free experiment revealed that KW-2189 bound to DNA more efficiently in the presence of HAC2 than HAC2/0.1 cell extract. It was concluded that decreased intracellular carboxyl esterase activity in HAC2/0.1 cells caused decreased intracellular conversion of KW-2189 to its active form, thus producing resistance to KW-2189. The decreased conversion of CPT-11 to SN-38 in HAC2/0.1 cells might be explained by decreased carboxyl esterase activity.
Antibiotics, Antineoplastic↗
Unusual location of intracranial vagus neurinoma--case report.
A 55-year-old male presented with hearing disturbance and tinnitus in the left ear. Computed tomography (CT) and magnetic resonance imaging demonstrated a well-defined, homogeneously enhanced mass in the left cerebellomedullary cistern without extension close to the jugular foramen. A three-dimensional image reconstructed from thin-slice CT scans demonstrated that the mass was clearly separated from the jugular foramen. The mass lesion was totally removed surgically. At surgery the tumor was found to originate from one rootlet of the vagal nerve just after its exit from the medulla oblongata. The histological diagnosis was neurinoma. Intracranial neurinomas of the glossopharyngeal, vagal, or accessory nerve usually originate within or close to the jugular foramen. This unusual location made it difficult to achieve a correct preoperative diagnosis.
Cranial Nerve Neoplasms↗
[Usefulness of 3-dimensional image analysis of skull base lesions].
We have evaluated three-dimensional (3D) images of the skull base lesions for planning cranial base surgery. Fifty 3D images were reconstructed from computed tomographies (CT), and/or magnetic resonance (MR) images or MR angiographic images of 30 patients with skull base lesions. These images have provided useful information for pre-operative evaluation. The 3D image reconstructed from CT provides clear information concerning the bone. Conversely, the 3D image from MR images demonstrates soft tissue very clearly, and that from MR angiography provides a detailed description of the vasculature. For skull base lesions, it is essential to evaluate 3D images from the different modalities, especially CT scan and MR image.
Adolescent↗
[Transcranial magnetic stimulation of the accesory nerve--investigation of the site and mechanism of excitation in the cat].
The site where transcranial magnetic stimulation excites the accessory nerve was studied in 5 cats. Transcranial magnetic stimulation of the accessory nerve was recorded from the right trapezius. The accessory nerve was stimulated electrically at the C1 level, jugular tubercle and jugular foramen. The latencies of the compound muscle action potentials (CMAPs) for each portion were measured and compared with the magnetic response, which was coincidental with that of the jugular tubercle. The accessory nerve was then transected in steps distally from the C1 level, and CMAPs following magnetic stimulation were recorded at each step. The CMAPs disappeared following the nerve transection at the jugular tubercle. The results of both approaches in this study conclude that transcranial magnetic stimulation excites the accessory nerve at jugular tubercle. This stimulation site was anatomically coincidental with that of the facial nerve and trigeminal nerve in being right before the point where the nerve bends. Following the accessory nerve transection at the C1 level, the amputation stump was moved cranially, and CMAPs disappeared. CMAPs recorded after the accessory nerve was returned to its original position. These examinations suggested that sudden alteration of the traveling lie of the nerve participates in the mechanism of transcranial magnetic stimulation.
Accessory Nerve↗
[Resection of recurrent lung cancer].
The resected recurrent lung cancer cases were evaluated retrospectively. Out of 1,060 cases who received an operation due to primary lung cancer, 21 (1.8%) had recurrent lung cancer which were resected subsequently. Stage I adenocarcinoma cases were found to be most frequent at the first operation (76%). Out of 21 cases, 7 received completion pneumonectomy, 5 had lobectomy and 8 had limited operation, respectively. The 5-year survival rate after the second operation was 36.6% in all the cases. There was no statistical difference in survival rate between the lobectomy group (including completion pneumonectomy) and the limited operation group. There was statistical difference in survival rate between cases who received a second operation in a time span of 2 or more years after the first operation and the cases who received it in less than 2 years. Good prognosis can be expected after the resection of recurrent lung cancer, but further analysis would be required in evaluation of respiratory function as well as biological malignancy of the tumor.
Adenocarcinoma↗
[A case report of pyomyositis--early diagnosis and follow-up by MRI].
We report a rare case of pyomyositis in a 28-year-old Japanese woman who was not immunocompromised. She was admitted because of high fever, sore throat, and severe tenderness and swelling of the right calf. Redness, swelling, and tenderness indicated presence of acute inflammation in the calf. CT of the lower extremities demonstrated low density areas in the right soleus muscle and surrounding fascia with marked swelling, which were of high signal on T2 weighted images of MRI. There was no finding of abscess formation. A tentative clinical diagnosis of acute pyomyositis was made, and antibiotics therapy with a combination of fosfomycin and sulbactam/cefoperazone was started although the arterial blood culture was negative for bacteria. Associated acute tonsilitis was the most probable focus of pyomyositis. Antibiotics relieved her symptoms, and the inflammation subsided in several weeks. No surgical procedure was necessary. MRI taken tree weeks after the onset demonstrated abscess formation between the soleus and gastrocnemius muscles. Slight high intensity indicating scar formation remained in the area of the former abscess six weeks after the onset. MRI was very useful not only in making the early diagnosis but also in the follow-up of pyomyositis.
Adult↗
[Cooperative multicentric study on posttraumatic epilepsy].
A multicenter cooperative study was conducted to investigate factors influencing posttraumatic epilepsy (PTE) and to evaluate the prophylactic effect of anticonvulsants. A retrospective study of 102 PTE patients revealed the following typical clinical features: occurrence in young males, traffic accidents, contusion and/or cerebral hematoma. The latent period after the injury was longer in children. The percentage of EEG paroxysmal activity gradually increased as the generalized abnormality diminished. A retrospective-prospective study of 1998 patients who suffered a head injury between 1984 and 1988 was conducted till 1994. During the follow-up period, 62 patients (3.1%) developed PTE. The drop-out cases were excluded, and the 154 cases followed at least two years were analyzed. Statistical analysis of differences between patients with and without PTE suggested following factors: young, immediate early epilepsy (within 24 hours after injury; IMEE) and early epilepsy (within one week after injury). The risk with the highest relative risk rate was early epilepsy. Multiple regression analysis revealed that three factors, IMEE, early epilepsy and young age, contributed to the prediction of PTE. There was no significant difference in the percentage of patients having PTE in the group treated with anticonvulsants and the untreated group. Anticonvulsant treatment after head injury was unlikely to have a prophylactic effect on the development of PTE.
Adolescent↗
Identification of the exact stimulated site in transcranial magnetic stimulation of the facial nerve.
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Electric Stimulation↗
Characterization of an etoposide-resistant human ovarian cancer cell line.
Etoposide (VP-16) is one of the most important anticancer agents available and is used in many chemotherapeutic regimens. To characterize resistance to this drug, we established a VP-16-resistant human ovarian cancer cell line, SKOV3/VP, by continuous stepwise exposure of SKOV3 cells to VP-16. The degree of resistance to VP-16 of SKOV3/VP was about 25 times that of the parent cell line (SKOV3), and SKOV3/VP showed cross-resistance to teniposide, adriamycin, CPT-11, and vincristine. The accumulation of [3H]-VP-16 observed in SKOV3/VP cells was about half that seen in SKOV3 cells, and the accumulation of Adriamycin by this resistant cell line was also lower than that of its parent. Overexpression of neither the multidrug resistance gene mdr-1, the multidrug-resistance-associated protein (mrp) gene, nor P-glycoprotein was detected using reverse transcriptase-polymerase chain reaction analysis and flow cytometry with MRK-16, a monoclonal antibody against P-glycoprotein. The topoisomerase II activity of nuclear extracts from SKOV3/VP cells was lower than that from the parental cells, as was the amount of DNA topoisomerase II, demonstrated by immunoblotting. These results suggest that the mechanism responsible for the multidrug resistance of this cell line may be attributable to changes on its DNA topoisomerase II and to its reduced accumulation of the drugs as compared with the parental line SKOV3.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
In vitro and in vivo growth of B16F10 melanoma cells transfected with interleukin-4 cDNA and gene therapy with the transfectant.
In an attempt to develop the most effective cytokine gene therapy, we transfected mouse interleukin(IL)-2, mouse IL-4, and human IL-6 cDNAs into mouse melanoma cells, B16F10. Transfection with IL-4 cDNA decreased the tumorigenicity of B16F10 most strongly. We investigated whether gene therapy with IL-4-transfected B16F10 cells was possible. Flow-cytometric analysis showed that major histocompatibility complex class I and II expression in B16F10 and IL-4-cDNA-transfected B16F10 (B16F10-IL4) cells did not differ. Doubling times of B16F10 and B16F10-IL4 were 20.1 and 21.1 h respectively. The growth of B16F10 cells was retarded if C57BL/6 mice were inoculated with B16F10-IL4 at the contralateral sides. When 5 x 10(5) B16F10 cells were transplanted subcutaneously into the flanks of C57BL/6 mice, they all developed a tumor mass, whereas no tumor masses formed in those transplanted with B16F10-IL4 cells within 60 days. No nude, severe combined immunodeficient or beige mice were able to reject parental B16F10 or B16F10-IL4 cells, although, B16F10-IL4 tumor growth in all these immunodeficient mice was slower than that of B16F10. Therefore, we concluded that T and natural killer cells are necessary for rejection of B16F10-IL4 tumor cells.
Animals↗
Characterization of a taxol-resistant human small-cell lung cancer cell line.
Taxol is a novel anticancer agent with activity against a broad range of tumors. It has a unique ability to stabilize polymerized tubulin into microtubule bundles within the cell. We have established a taxol-resistant human small-cell lung cancer cell line (H69/Txl) by exposing H69 cells to stepwise increases in taxol concentration. The resistance of H69/Txl cells to taxol was 4.7-fold that of the original H69 cells: the IC50 values for H69 and H69/Txl were 113.7 +/- 56.54 nM and 538.7 +/- 214.7 nM by the tetrazolium dye assay, respectively. Removal of the drug from the medium resulted in a 38% decrease in the growth rate of H69/Txl as compared with that in the presence of 30 nM taxol, suggesting that the growth of H69/Txl was partially dependent on taxol. H69/Txl showed higher sensitivity to vinca alkaloids such as vindesine, vincristine and vinblastine than the parental H69. There was no significant difference in intracellular [3H]taxol content between H69 and H69/Txl cells. No MDR-1 mRNA was detected in H69/Txl by the reverse transcription polymerase chain reaction. There was no significant difference of total and polymerized tubulin content between H69 and H69/Txl cells. Altered mobility of one of the alpha-tubulin isoforms in H69/Txl was revealed by using isoelectric focusing and Western blotting with anti-alpha-tubulin antibody. In H69, two alpha-tubulin isoforms were observed, whereas three were evident in H69/Txl, two of them comigrating with the isoforms of H69 and the other being more acidic. We observed the increased acetylation of alpha-tubulin in H69/Txl cells as compared with that in H69 cells. The acetylation of alpha-tubulin may be responsible for the taxol resistance and/or taxol-dependent growth of H69/Txl.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
A novel antitumor antibiotic, KW-2189 is activated by carboxyl esterase and induces DNA strand breaks in human small cell lung cancer cells.
KW-2189 has been selected as a lead compound for clinical trial among duocarmycin derivatives with structural similarity to CC-1065, a cyclopropylpyrroloindole. The purpose of this study was to examine the DNA-binding potency and the mechanisms of cytotoxicity of KW-2189. In order to analyze DNA-binding activity of KW-2189, plasmid pBR322 was treated with KW-2189 with or without pretreatment with carboxyl esterase, which we demonstrated to be an activating enzyme, and the products were examined by agarose gel electrophoresis and restriction enzyme analysis. Cytotoxic activity was examined by exposing a human small cell lung cancer cell line, NCI-H69 to KW-2189 with or without carboxyl esterase. Alkaline elution was performed to examine whether KW-2189 induces DNA strand breaks. DNA treated with KW-2189 and carboxyl esterase migrated faster than KW-2189-treated DNA, which migrated at the same rate as untreated DNA. In addition DNA treated with esterase-activated KW-2189 was protected from digestion by some restriction enzymes. KW-2189 showed concentration- and time-dependent growth inhibitory effect with IC50 values (drug concentration required for 50% growth inhibition) of 58 nM (96 h) to 1900 nM (1 h) in H69 cells. The IC50 values of 4-h exposure of H69 to KW-2189 with 0, 26, 130, 650 mU/ml carboxyl esterase were 460, 120, 30, and 7 nM, respectively. Time-dependent enhancement of cytotoxicity by carboxyl esterase was also observed. KW-2189 induced DNA strand breaks in H69 cells in a concentration-dependent manner around the IC50 value. We conclude that 1) KW-2189 is activated by carboxyl esterase to its active form(s), 2) activated KW-2189 has a stronger DNA-binding activity and cytotoxicity than KW-2189, 3) DNA cleavage is one of the major mechanisms of KW-2189-mediated cytotoxicity.
Antibiotics, Antineoplastic↗
Genistein, a tyrosine kinase inhibitor, decreased the affinity of p56lck to beta-chain of interleukin-2 receptor in human natural killer (NK)-rich cells and decreased NK-mediated cytotoxicity.
Intracellular signal transduction has been reported to be triggered by phosphorylation of interleukin-2 (IL-2) receptor by IL-2. In order to clarify the effect of tyrosine phosphorylation of IL-2 receptors on cell-mediated cytotoxicity by natural killer (NK) cells, we studied the effect of a tyrosine kinase inhibitor, genistein, on the lethal effects of NK-rich cells for K562 cells. Exposure of NK-rich cells to IL-2 (100 U/ml) for 3 days increased their cytotoxicities against K562 cells. The effect was reduced in the presence of 10 micrograms/ml of genistein. Samples immunoprecipitated by anti-IL-2R beta antibodies were prepared from NK-rich fractions with or without exposure to IL-2 and/or genistein. Coprecipitated proteins with 75, 65, and 56 kDa were detected with an antiphosphotyrosine antibody. The amount of phosphorylated tyrosine residues of 56-kDa protein, which was predominantly detected in NK-rich cells, was remarkably increased by IL-2 treatment. The enhanced phosphorylation of 56-kDa protein was reduced by the presence of genistein. These results suggested that IL-2 increased tyrosine phosphorylation and the affinity to IL-2R beta of the 56-kDa proteins in NK-rich cells. Immunoprecipitated samples by anti-IL-2R beta were reblotted with anti-p56lck antibody and revealed that the 56-kDa protein was identified to be p56lck. The increase of coprecipitated p56lck with anti-IL-2R beta antibody by the treatment with IL-2 suggested that the affinity of p56lck to IL-2R beta was increased by IL-2 in NK-rich cells. The amount of coprecipitated p56lck with IL-2R beta was reduced in the sample exposed to genistein. The affinity of p56lck to IL-2R beta was considered to be regulated by IL-2-induced tyrosine phosphorylation. Our results demonstrated a potential role for tyrosine kinase, p56lck, in the signaling events that regulate the cytotoxicity by NK-rich cells.
Cytotoxicity, Immunologic↗