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I Hayata

Publications and source records attributed to I Hayata.

At least 19 recordsLinked to original sources
PubMed1992

A high resolution chromosome image processor for study purposes, NIRS-1000:CHROMO STUDY, and algorithm developing to classify radiation induced aberrations.

Since 1989 we have promoted a project to develop an automated scoring system of radiation induced chromosome aberrations. As a first step, a high resolution image processing system for study purposes, NIRS-1000:CHROMO STUDY, has been developed. It is composed of: (1) CHROMO MARKER whose main purpose is to mark on images to make image data base, (2) CHROMO ALGO whose purpose is algorithm development, and (3) METAPHASE RANKER whose purposes are metaphase finding and ranking with a high power objective lens. However, METAPHASE RANKER is presently under development. The system utilizes a high definition video system so as to realize the best spatial resolution that is achievable with an optical microscope using an objective lens (x 100, numerical aperture 1.4). The video camera has 1024 effective scan lines to realize 0.1 microns sampling on a specimen. The system resolution achieved on the hard copy is less than 0.3 microns on a specimen. A preliminary algorithm has been developed to classify the aberrations on the system using projection information of gray level. The preliminary test results on excellent 10 metaphases show that the correct classification ratio is 92.7%, that the detection rate of the aberrations is 83.3% and that the false positive rate is 6.1%.

Algorithms↗
PubMed1992

Robot system for preparing lymphocyte chromosome.

Towards the automatization of the scoring of chromosome aberrations in radiation dosimetry with the emphasis on the improvement of biological preparations, the conventional culture and harvesting method was modified. Based on this modified method, a culture and harvest robotic system (CHROSY) for system (CHROSY) for preparing lymphocyte chromosome was developed. The targeted points of the modification are as in the following. 1. Starting culture with purified lymphocytes in a fixed cell number. 2. Avoiding the loss of cells in changing the liquids following centrifugalization. 3. Keeping the quantity of the liquids to be applied to the treatments of cells fixed. 4. Building a system even a beginner can handle. System features are as follows. 1. Operation system: Handling robot having 5 degrees of freedom; a rotator incubator with an automatic sliding door; units for setting and removing pipette tips; a centrifuge equipped with a position adjuster and an automatic sliding door; two aluminum block baths; two nozzles as pipettes and aspirators connected to air pumps; a capping unit with a nozzle for CO2 gas; a compressor; and an air manipulated syringe. 2. Control system: NEC PC-9801RX21 with CRT; and program written in Basic and Assembly languages on MS-DOS. It took this system 2 hours and 25 minutes to harvest 2 cultures. A fairly good chromosome slide was made from the sample harvested by CHROSY automatically.

Chromosomes↗
PubMed1992

[Progress on chromosome image analysis systems].

This is a review of the development of automated systems of chromosome analysis. The history of its development reflects the progress of image analysis systems and that of method of chromosome analysis in man. It can be chronologically summarized as follows. Period I: 1960s-early 1970s when chromosome banding methods were discovered. Period II: early 1970s-early 1980s when the first image analysis system of chromosome was commercialized. Period III: early 1980s-present. Research and development of the automated analysis system of radiation induced chromosome aberrations, conducted under the national project of cross-over research on underlying technology of nuclear energy, Science and Technology Agency, Japan, is briefly reported.

Chromosomes↗
PubMed1990

Enhancement of an anti-tumor effect of interferon by dipyridamole in established human malignant melanoma cell lines.

Enhancement of the anti-proliferative effect of human interferon (HuIFN) preparations (alpha, beta and gamma) by dipyridamole was detected in a human malignant melanoma cell line, MM-ICB, which we originally established. Cell growth was inhibited by HuIFN alone, but a marked increase in inhibition was noted in vitro and in vivo when dipyrydamole was added. Cellular DNA synthesis, as determined by 3H-deoxythymidine incorporation into the acid-insoluble cellular fraction, was more inhibited by combined treatment than by any of the agents used alone. Two other melanoma cell lines that we established, MM-2CB and MM-3CB, also exhibited sensitivity to combined treatment both in vitro and in vivo. Furthermore, the HMV-I and SEKI melanoma cell lines were susceptible to the combination. Even non-cytotoxic concentrations of dipyridamole could enhance the effect of HuIFN on MM-ICB, MM-2CB, and SEKI cells.

Aged↗
PubMed1989

Progression and selection in heterogeneous tumor composed of androgen-responsive Shionogi carcinoma 115 and its autonomous subline (Chiba subline 2).

Shionogi Carcinoma 115 (SC 115) is an androgen-dependent mouse tumor, and Chiba subline 2 (CS 2) is its androgen-independent subline which differs from SC 115 in cell size, amount of androgen receptors, and karyotype. To shed light on the mechanism of clonal selection of androgen-independent tumors, mixed tumors with SC 115 and CS 2 were prepared, and growth of these tumors was examined in vivo and in vitro. When the mixed tumor was transplanted in mice, CS 2 showed a predominant growth over SC 115. In a culture of mixed tumor cells, however, CS 2 showed no selective growth advantage. The suppressive interaction which occurred in vivo was due neither to transferable substances, nor to some immunological factor(s). It may be, at least partially, attributable to necrosis formation in SC 115, which developed with an increase in the size of the tumor.

Androgens↗
PubMed1989

Predominant use of a particular alpha-chain in suppressor T cell hybridomas specific for keyhole limpet hemocyanin.

We isolated and sequenced the rearranged genomic variable (V) and joining (J) gene segments of T cell receptor alpha-chain gene from two independent keyhole limpet hemocyanin (KLH)-specific suppressor T cell (Ts) hybridomas (BW5147 x C57BL/6 KLH-Ts). These nucleotide sequences were compared with those of germline DNA from kidney and also with cDNA of alpha-chain (VJ alpha 281) previously isolated from Ts hybridoma (34S-281) with KLH/H-2b Ts activity. The entire V alpha and J alpha sequences of all three Ts hybridomas were exactly identical and were encoded by the germline V alpha and J alpha gene segments without any mutations, except for 2-nucleotide deletions from both the 3' end of V alpha and 5' end of J alpha gene segments, respectively, and a 1-nucleotide (guanine) insertion in the junctional (N) region which was not encoded by the germline gene. Six additional KLH-Ts hybridomas, further analyzed, also possessed the same alpha-chain, indicating the preferential usage of the particular alpha-chain in these hybridomas. As chromosome analysis demonstrated a different pattern in each clone, these hybridomas appear to be independent. More surprisingly, 0.5-1.5% of the total functional T cell alpha-chain mRNA in the thymus and spleen of unprimed C57BL/6 mice was found to be of this particular alpha-chain. These results suggest that the repertoire of KLH-Ts is strictly limited.

Amino Acid Sequence↗
PubMed1989

Effects of recombinant human granulocyte colony stimulating factor (rG-CSF) on murine myeloid leukemia: stimulation of proliferation of leukemic cells in vitro and inhibition of development of leukemia in vivo.

We have established an experimental murine myeloid leukemia model and investigated the effects of recombinant granulocyte colony stimulating factor (rG-CSF) on myeloid leukemia in vitro and in vivo. rG-CSF stimulated colony formation by the leukemic cells in semisolid agar medium, and exponential growth of the clonogenic cells in suspension medium. Thus, rG-CSF was able to stimulate both the differentiation and self-renewal processes of the leukemic stem cells in vitro. However, 14 consecutive daily injections of rG-CSF prolonged the mean survival time of the mice implanted with the leukemic cells. This effect of rG-CSF was accompanied by a delay in the emergence of the blast cells in peripheral blood and by a decreased blast population in the spleen, suggesting that development of leukemia was suppressed in the rG-CSF-treated-mice. The prolongation of the survival time by rG-CSF was more evident when rG-CSF was administered in therapeutic combination with cyclophosphamide. These results indicate that the effect of rG-CSF on the development of leukemia is not exactly predicted from in vitro experiments.

Animals↗
PubMed1988

Radiosensitivity of late recurrences following radiotherapy of murine fibrosarcomas.

Radiosensitivity of late recurrent tumors which emerged after radiotherapy was investigated. Tumors observed were fibrosarcomas. Recurrences emerged in the irradiated area approximately 200 days after a 50% tumor control dose of radiation of 60Co gamma rays or mixed irradiation with fast neutrons and gamma rays. The recurrent and radiation-induced tumors were differentiated by karyotype analysis. Once transplanted into fresh mice, the recurrent tumors grew more slowly than the original tumor. Tumorigenicity of the late recurrences was lower than that of the original tumor. Radiosensitivity of the late recurrences, which was examined using methods to assess control, tumor growth delay, and colony forming assays, was significantly higher than that of the original tumor. D0 values of hypoxic tumor cells were significantly smaller in two of the three recurrences compared to the original tumor. Oxic cells, when irradiated in vitro, also showed smaller D0 values for the recurrent tumors than the original tumor. Hypoxic cell fractions were between 0 and 14% in the late recurrences and 10% in the original tumor. These results are consistent with the hypothesis that radiotherapy causes mutation of tumor cells which results in increased radiosensitivity of surviving tumor cells.

Animals↗
PubMed1987

Myeloproliferative disorder due to abnormal production of hematopoietic stimulators.

A new kind of myeloproliferative disorder (L-8313) has been discovered. It was transplantable into syngeneic mice with spleen cells. The mice showed hepato-splenomegaly with a marked leukocytosis and anemia 3 weeks after transplantation of L-8313 cells. The number of GM-CFU and CFU-S per spleen increased to more than 40 times normal. The results of chromosomal and PGK analysis demonstrated that these increased stem cells were of host origin. Both the culture medium of the spleen cells and the serum from L-8313 bearing mice showed high levels of IL-3, BPA and CSF. Consequently, hematopoietic cells of the host mice underwent remarkable proliferation in response to these stimulating factors when L-8313 cells were transplanted. We also have been successful in establishing an in-vitro cell line and have maintained it for over one year. The phenotype of L-8313 cells was Thy 1.2 positive. Some L-8313 cells showed a positive acid phosphatase reaction but the cytochemical character of myeloid lineage was not observed. Therefore, L-8313 is considered to be a T-cell derived hematopoietic regulatory cell neoplasm with the ability to produce several hematopoietic stimulating factors.

Anemia↗
PubMed1987

Androgen receptor, testosterone uptake and karyotype in androgen-dependent mouse tumor (SC 115) and its androgen-independent subline (CS 2).

Content of androgen receptor, retention of injected testosterone and karyotype of SC 115, androgen-dependent tumor, were compared with those of CS 2, an androgen-independent subline derived from SC 115. Although Bmax was less than that of SC 115, androgen receptor was present in the cytosol and the nuclear extract from CS 2. To examine the ability for androgen retention, a large amount of testosterone was injected into tumor-bearing mice, and the amount of androgen in the crude nuclear and postnuclear fractions of tumors was compared. In both fractions, retention of injected androgen was higher in the SC 115 than in the CS 2. Since most of the injected testosterone was not metabolized in the tissues and the injection of testosterone 5 alpha-reductase inhibitor showed no significant influence on the growth rate of the SC 115, intracellular active androgen was assumed to be testosterone in these tumor cells. As the CS 2 was tetraploid, the androgen independency of the CS 2 seems to be related to chromosomal changes.

Animals↗
PubMed1986

Nature of leukemic stem cells in murine myelogenous leukemia.

We investigated the nature of myelogenous leukemic stem cells in mice. L-8057, a megakaryoblastic leukemia cell line used in this study, produces in vivo and in vitro colonies. By means of typical chromosomal aberrations in L-8057, one can conveniently detect the origin of the cells in each colony derived from a leukemic stem cell. Direct evidence of whether cells from each colony had leukemogenicity in recipient mice was successfully obtained by the colony transplantation assay. Both leukemic colony-forming unit-spleen (L-CFU-s) and leukemic colony-forming unit-culture (L-CFU-c) in L-8057 may have belonged to the same differentiating stage in the stem cells because of their similar radiosensitivity, although some parts of the L-CFU of L-8057 seemed to have lost their capability to regenerate L-CFU-s when the cells were plated in dishes. This leukemic stem cell preserves high self-renewal ability in vitro after 10 passages. In addition, in vitro colony formation by this leukemic cell during the above course of serial passages did not require any additional exogenous stimulators. The same sort of trials have been made on other types of leukemias. Leukemic stem cells showed remarkable variety in their response to stimulating factors and in their self-renewal activity, which suggests that they may have consisted of heterogeneous populations.

Animals↗
PubMed1985

Increased radiosensitivity of a recurrent murine fibrosarcoma following radiotherapy.

The radiosensitivity of a recurrent tumor following local radiotherapy was investigated in order to determine whether or not the cell(s) surviving radiotherapy are radioresistant. A tumor which recurred in a C3Hf/HeMsNrs male mouse 200 days after local irradiation of a transplanted syngeneic fibrosarcoma was examined. The recurrent tumor showed the same biological properties, such as karyotype, histological features, and ability to produce colony stimulation factor, as the original tumor. On the other hand, the tumor control dose and the D0 value of the recurrent tumor were much smaller than those of the original tumor. Moreover, the recurrent tumor grew more slowly than the original tumor. It was concluded that initial radiotherapy might have changed the intrinsic radiosensitivity of the original tumor cells.

Animals↗
PubMed1983

Reconfirmation of indirect induction of radiogenic lymphomas using thymectomized, irradiated B10 mice grafted with neonatal thymuses from Thy 1 congenic donors.

An experiment was conducted to reexamine earlier observations that lymphomas could develop from lymphocytes present in nonirradiated thymuses grafted into thymectomized, fractionally (170 R, 4 doses) irradiated mice by using B10. Thy 1 congenic donor-host combinations. The results indicated that: (a) 37 of 91 thymectomized, fractionally irradiated B10. Thy 1.2 mice which were grafted s.c. with 7-day-old thymuses from B10. Thy 1.1 donor mice had developed frank lymphomas between 90 and 270 days after thymus grafting; (b) 28 of 37 lymphomas developed in this group were typed individually with respect to Thy 1 alloantigens by the cytotoxicity assay using monoclonal anti-Thy 1.1 (T-11-D7) and anti-Thy 1.2 (F7D5) antibodies plus complement. It was shown that 21 thymic lymphomas (75%) had originated from lymphocytes of the nonirradiated thymus grafts and 5 tumors (17.9%) from cells of the irradiated hosts; 2 thymic lymphomas (7.1%) manifested no Thy 1 antigens; (c) lymphoma cells originated from both nonirradiated thymus grafts and irradiated hosts possessed chromosome abnormalities, which were mainly numerical changes of some chromosomes or polyploidizations.

Animals↗
PubMed1979

Partial deletion of chromosome No. 2 in myelocytic leukemias of irradiated C3H/He and RFM mice.

Chromosomes of mouse myelocytic leukemias that developed in 7 irradiated mice, 3 C3H/He males, 1 RFM female, and 3 RFM males were analyzed with chromosome-banding techniques. Chromosomes No. 2 were partially deleted in 6 of the 7 mice. Although the deleted No. 2 chromosomes varied in size in the 6 mice, one common characteristic was noted in all these deletions: A segment lying between a certain band in the region 2C and a band in the region 2E, including the whole region 2D, was missing. Another consistent abnormality was an addition or a loss of the Y-chromosomes in the fraction of cells in all 6 males. In addition to these consistent abnormalities, various chromosomes had structural abnormalities. The RFM female, which did not have the abnormal No. 2 chromosome, had abnormalities in chromosomes No. 3, 4, 11, 12 and 15 and in the X-chromosome. Of the 20 chromosome pairs, only such chromosomes as No. 1, 5, 8, 14, 17, and 19 and the Y-chromosome did not have the structural abnormalities. The possible role of the partial deletion of the No. 2 chromosome was considered in relation to the development of mouse myeloid leukemias.

Animals↗
PubMed1979

Chromosome markers in 12 inbred strains of the Norway rat, Rattus norvegicus.

Morphological details of metaphase chromosomes were compared among 12 inbred strains of rats (Rattus norvegicus) by means of conventional Giemsa staining and by a sequential Q- and C-banding method. Inter-strain variations were found in seven pairs, as identified on the basis of size differences in the short arms and/or satellites of chromosomes 3 and 12 and the X chromosome and in the centromeric C-bands of chromosomes 4, 5, 7, and 9. All pairs were homomorphic in the inbred strains, while F1 hybrids between two inbred strains showed certain heteromorphic pairs expected from the parents. These chromosome markers appear to be useful for characterization of inbred strains as well as for various genetic studies, including linkage analyses.

Animals↗
PubMed1979

Development of androgen-independent spindle cell tumors from androgen-dependent medullary Shionogi carcinoma 115 in androgen-depleted nude mice.

When Shionogi carcinoma 115 (SC115, undifferentiated medullary carcinoma showing compact cell pattern and containing androgen receptor) was transplanted into male and female DS mice, it grew only in males. In contrast to this strict androgen dependency in DS hosts, tumors composed of spindle-shaped cells appeared in more than 80% of cases when SC115 tumor was inoculated into female or castrated male nude athymic (BALB/c-nu/nu) recipients. These spindle cell tumors neither contained cytosol androgen receptor nor showed biologically defined androgen dependency. As spindle cell tumors could be serially transplanted in DS mice but not in BALB/c-+/+ mice and as the original SC115 (medullary carcinoma showing a compact cell pattern) tumor and the spindle cell tumor had many identical chromosome abnormalities, these two types of tumors seem to have a common origin in spite of their morphological, biochemical, and biological differences. Since spindle cells could not be detected histologically in SC115 tumors maintained in intact male DS mice, the present results seem to suggest that SC115 cells may change their morphological, biochemical, and biological characteristics within one passage in androgen-depleted nude athymic mice.

Androgens↗