Biomedical subjects

K Morishita

Publications and source records attributed to K Morishita.

At least 19 recordsLinked to original sources

PubMed1992

Activation of EVI1 gene expression in human acute myelogenous leukemias by translocations spanning 300-400 kilobases on chromosome band 3q26.

Retroviral activation of Evi-1 gene expression is one of the most common transforming events in murine myeloid leukemias. To evaluate the role of the EVI1 gene in human acute myelogenous leukemia (AML), leukemic blasts or cell lines from 116 patients were examined. In eight patients the EVI1 gene was expressed and all but one had cytogenetically detectable translocations of chromosome 3q26 where the EVI1 gene has been localized. To identify breakpoints, a restriction map that spans 1700 kilobases (kb) of the EVI1 locus was developed by pulsed-field gel electrophoresis. In one case, t(3;3)(q21;q26), a rearrangement was localized to 170-330 kb 5' of the gene. In a second case, t(3;3)(q21;q26), there was a rearrangement 13 kb 5' of the gene. This rearrangement was cloned and shown to be due to the fusion of sequences from 3q21-22 with the EVI1 locus. In the third case, ins(3)-(q21q25q27), there was a rearrangement that mapped 150 kb downstream from the 5' end of the gene.

Base Sequence↗

PubMed1992

Expression and characterization of kinase-active v-erbB protein using a baculovirus vector system.

The v-erbB gene is an oncogene of the avian erythroblastosis virus encoding a protein that is a truncated version of the epidermal growth factor receptor. The v-erbB protein was expressed alone or as polyhedrin-erbB fusion proteins using the Bombyx mori nuclear polyhedrosis virus vector. The expression level of the fusion protein whose polyhedrin portion consisted of only 8 amino-terminal amino acids was more than ten times higher than that of the non-fusion protein. Studies with tunicamycin showed that the recombinant v-erbB proteins were glycosylated. The recombinant protein autophosphorylated tyrosine residues, and phosphorylated a synthetic tyrosine-containing peptide and lipocortin I. These observations indicate that functional v-erbB protein can be expressed in silkworm-derived cells, and furthermore, that this system can be used for large-scale production.

Amino Acid Sequence↗

PubMed1992

Expression of the Evi-1 zinc finger gene in 32Dc13 myeloid cells blocks granulocytic differentiation in response to granulocyte colony-stimulating factor.

Expression of the Evi-1 gene is frequently activated in murine myeloid leukemias by retroviral insertions immediately 5' or 90 kb 5' of the gene. The Evi-1 gene product is a nuclear, DNA-binding zinc finger protein of 145 kDa. On the basis of the properties of the myeloid cell lines in which the Evi-1 gene is activated, it has been hypothesized that its expression blocks normal differentiation. To explore this proposed role, we have constructed a retrovirus vector containing the gene and examined its effects on an interleukin-3-dependent myeloid cell line that differentiates in response to granulocyte colony-stimulating factor (G-CSF). Expression of the Evi-1 gene in these cells did not alter the normal growth factor requirements of the cells. However, expression of the Evi-1 gene blocked the ability of the cells to express myeloperoxidase and to terminally differentiate to granulocytes in response to G-CSF. This effect was not due to altered expression of the G-CSF receptor or to changes in the initial responses of the cells to G-CSF. These results support the hypothesis that the inappropriate expression of the Evi-1 gene in myeloid cells interferes with the ability of the cells to terminally differentiate.

Animals↗

PubMed1992

Clinical management of boric acid ingestion: pharmacokinetic assessment of efficacy of hemodialysis for treatment of acute boric acid poisoning.

Seven hours after suicidal ingestion of about 21 g of boric acid, a 26-year-old female admitted to our hospital in a state of slightly impaired consciousness, with frequent vomiting, shivering, fever and skin flush. Immediately, gastric lavage, followed by administration of activated charcoal and laxative (MgSO4), was performed. In order to ensure her urination, fluid infusion therapy was conducted with the aid of diuretics (furosemide). Since the serum concentrations of boric acid was very high, hemodialysis was carried out twice during the first 39 h. She responded well to the above mentioned treatment and was discharged 12 d post-admission without any sequelae. The concentrations of boric acid in serum and urine were measured in appropriate intervals with our modified Miyamoto's method, and the pharmacokinetics of boric acid were analyzed. The concentration of boric acid in serum and urine at the beginning of treatment was 465 micrograms/ml and 3.40 mg/ml, respectively. The half-life of boric acid in serum was 13.46 h, whereas it was shortened to 3.76 h during hemodialysis. The total body clearance was 0.99 l/h, while it increased to 3.53 l/h by hemodialysis. The additional removal of boric acid by hemodialysis was estimated to be about 5 g. It was concluded that the hemodialysis was very useful in the treatment of boric acid poisoning, because it accelerated the elimination of boric acid about four times faster than with conventional treatment.

Acute Disease↗

PubMed1991

Effect of ticlopidine and other antithrombotics on the venous thrombosis induced by endothelial damage of jugular vein in rats.

A venous thrombosis model was produced by vessel wall damage generated by freezing a small segment of the rat jugular vein. The process of thrombus formation was investigated by electron microscopic study. Ultrastructural studies demonstrated that the initiation of thrombus formation could be deendothelialization caused by freezing of vessel. When blood flow was reestablished, platelets adhered to subendothelium within 1 min. Then platelets aggregated on the adhering platelets, and fibrin net was formed. Finally, thrombi composed predominantly of fibrin and red blood cells with platelet aggregates and leukocytes were generated. An anti-platelet agent, ticlopidine, revealed a potent antithrombotic effect in this model. Because ticlopidine decreased the number of platelet aggregates, reduced the size of aggregates, and inhibited platelet degranulation, it is conceivable that platelet aggregation in early phase of thrombus formation plays a crucial role even in venous thrombosis model. A synthetic thrombin inhibitor, argatroban, also showed a potent antithrombotic effect, but a thrombolytic agent, urokinase, was less effective. In conclusion, this model is both platelet- and coagulation-dependent.

Animals↗

PubMed1991

Effects of volume loading on left atrial systolic time intervals.

The effects of volume loading on the left atrial preejection period (LAPEP) and left atrial ejection time (LAET) were examined in 24 patients with various heart diseases using pulsed Doppler echocardiography. In response to volume loading, the left atrial dimension before atrial contraction significantly increased from 30.6 mm +/- 5.8 mm to 32.4 mm +/- 5.4 mm and the change in the left atrial dimension during atrial contraction tended to increase. The peak velocity in the atrial contraction phase significantly increased from 58 cm/s +/- 14 cm/s to 63 cm/s +/- 13 cm/s, and the integral of the atrial contraction phase tended to increase. LAPEP significantly decreased from 114 ms +/- 16 ms to 104 ms +/- 14 ms and LAET significantly decreased from 128 ms +/- 15 ms to 124 +/- 12 ms. The relation between LAET and left ventricular end-diastolic pressure, and that between LAPEP and mean pulmonary capillary wedge pressure, shifted downward to the right after volume loading. Thus, left atrial ejection is augmented by volume loading according to the Frank-Starling mechanism, while LAPEP decreases due to an increase in preload and LAET decreases due to an increase in afterload.

Atrial Function, Left↗

PubMed1991

Fuzzy realization in clinical test database system.

To be able to obtain useful information from medical data easily and quickly in daily use for the hospital staff, it is necessary to construct a proper database system using methodologies suitable for the nature of medical data. From this viewpoint we have developed the clinical test relational database system at Hiroshima University Hospital, which provides the functions of easy on-line retrieval and statistical analysis. Fuzzy query processing based on the fuzzy set theory is adopted in the system. This enables us to use natural linguistic representation and makes it easy to introduce medical knowledge representation. Fuzzy set approaches turn out to be superior in clinical evaluation of laboratory data to the ordinary clear-cut definition of normalcy.

Artificial Intelligence↗

PubMed1991

Characterization of v-sis protein expressed in silkworm larvae using the Bombyx mori nuclear polyhedrosis virus vector.

The v-sis oncogene of simian sarcoma virus encodes a protein which is homologous to the human platelet-derived growth factor B-chain. The v-sis protein undergoes a series of processing steps including dimer formation and proteolytic digestion to generate several molecular sizes of the protein. Two of these v-sis proteins were expressed alone or as polyhedrin-sis fusion proteins using the Bombyx mori nuclear polyhedrosis virus vector. The polyhedrin-sis fusion proteins contained a collagenase-sensitive site at the junction. The expression levels of the fusion proteins whose polyhedrin portions consisted of only 8 amino-terminal amino acids were 3-4 times higher than those of non-fusion proteins. One of these fusion proteins was expressed in silkworm larvae and the v-sis protein was isolated from the fusion protein by collagenolysis followed by chromatography. Because the purified v-sis protein exhibited the same molecular size on SDS-polyacrylamide gels under reducing and non-reducing conditions, it was concluded to be monomeric in structure. It possessed chemotactic activity but lacked mitogenic activity. In addition, a small amount (approximately 1%) of monomeric v-sis protein was converted in vitro to the mitogenically active v-sis protein, which could be a homo-dimer.

Amino Acid Sequence↗

PubMed1991

Solitary metastatic ovarian carcinoma of the spleen: a case report.

Carcinomatous metastatic involvement of the spleen usually indicates a widespread malignant disease. Solitary metastatic lesions in the spleen are exceedingly rare. The literature contains fewer than 16 cases. In this paper we report a case of a solitary metastatic lesion of the spleen arising from a serous cystadenocarcinoma of the ovary 5 years after the initial operation. A splenectomy was performed followed by smooth postoperative course.

Aclarubicin↗

PubMed1991

[The development and the applications of clinical laboratory system by using of fuzzy set theory].

For clinical laboratory data to be used effectively in wide areas of clinical medicine, it must be stored in proper database systems. Moreover, the system must be installed in the hospital information system for easy access by the hospital staff in their routine activities. From this standpoint we have developed a clinical test relational database system at Hiroshima University Hospital, which provides the functions of easy on-line retrieval and statistical analysis. We have also improved the functions to incorporate fuzzy query processing based on the theory of fuzzy sets. This system enables us to use natural linguistic representation and leads to a medical knowledge-base.

Clinical Laboratory Information Systems↗

PubMed1990

The influence of intra-arterial infusion of cisplatinum and bleomycin on the cell cycle in uterine cervical carcinoma.

We examine the influence of CDDP and BLM on the cell cycle in the patients with uterine cervical cancer. Specimens were obtained from 8 cases of local recurrent uterine cervical cancer which were treated with CDDP and BLM bolus infusions into bilateral internal iliac arteries. Except for two patients who showed no response according to the criteria of the National Cancer Institute (NCI), the other patients exhibited partial response (PR). The concentrations of BLM and CDDP were 0.83 +/- 0.05 microgram/g and 2.9 +/- 0.54 microgram/g in the tumor tissues 24 hours after treatment. The peak values of the drug concentrations in serum, which were observed at 30 min for both drugs, were 0.92 +/- 0.12 microgram/ml BLM and 1.02 +/- 0.65 microgram/ml CDDP and the concentrations of the drugs decreased rapidly in the serum. The higher levels of CDDP and BLM were retained in the tumor tissue 24 hours after treatment, than in serum. Changes in DNA content determined by flow cytometry, revealed that proliferation indices (PI) increased after treatment only in PR cases. The mean increase in the ratio was 0.10 +/- 0.015. The BrdU staining showed that cells in S phase increased in tumor tissues. Labeling indices increased from 0.10 +/- 0.04 to 0.18 +/- 0.05. These results suggested that cell synchronization was induced by CDDP and BLM in vivo. PI seemed a useful parameter for measuring drug sensitivity.

Aged↗

PubMed1990

Phenotypes and mechanisms in the transformation of hematopoietic cells.

Interleukin 3 (IL-3) is a growth factor that supports the proliferation of early hematopoietic stem cells, as well as cells that are committed to a variety of the myeloid lineages. The mechanisms by which IL-3 functions have been studied through the use of a series of IL-3-dependent cell lines isolated from myeloid leukemias or long-term bone marrow cultures. A variety of studies have implicated tyrosine phosphorylation in IL-3 signal transduction. One of the substrates of phosphorylation is a 140 kDa, IL-3-binding protein that is speculated to be the biologically relevant IL-3 receptor. IL-3, through tyrosine phosphorylation, supports viability and growth through the regulation of transcription of a series of genes including c-myc and c-pim-1. The c-myc gene contributes to viability, in part, by regulating the transcription of the ornithine decarboxylase gene. The role of growth factors in differentiation is less clear. By studying IL-3-dependent myeloid leukemia cell lines, two genes have been identified whose altered expression is associated with blocking the ability of the cells to differentiate. The c-myb gene is a nuclear DNA binding protein that has been implicated in myeloid transformation in a number of systems. The Evi-1 gene is a novel gene of the zinc finger family of transcriptional activators. Possible mechanisms by which these genes interfere with normal differentiation are discussed.

Animals↗

PubMed1990

Identification, nuclear localization, and DNA-binding activity of the zinc finger protein encoded by the Evi-1 myeloid transforming gene.

Activation of the Evi-1 zinc finger gene is a common event associated with transformation of murine myeloid leukemias. To characterize the gene product, we developed antisera against various protein domains. These antisera primarily detected a 145-kilodalton nuclear protein that bound double-stranded DNA. Binding was inhibited by chelating agents and partially restored by zinc ions.

Blotting, Western↗

PubMed1990

The Evi-1 zinc finger myeloid transforming gene is normally expressed in the kidney and in developing oocytes.

Activation of the Evi-1 zinc finger gene is commonly associated with the transformation of murine leukemias and is involved in some cases of human AML involving rearrangements at chromosome 3q25. To determine the normal function of the gene, we have looked for expression in a variety of cell lines and tissues. The predominant sites of expression of the gene are in the kidney and ovary. In the kidney, expression is localized to the renal tubules in the corticomedullary junction. In the ovary, high levels of the Evi-1 protein are found in the cytoplasm of developing oocytes. The latter result suggests a potential role for the Evi-1 gene product in early oocyte development.

Animals↗

PubMed1990

The human Evi-1 gene is located on chromosome 3q24-q28 but is not rearranged in three cases of acute nonlymphocytic leukemias containing t(3;5)(q25;q34) translocations.

The murine Evi-1 gene encodes a protein that has multiple 28-amino acid repeats containing the consensus sequence found in the zinc finger domains of many transcriptional regulatory proteins. Activation of the expression of the Evi-1 gene is frequently found in murine myeloid leukemias and leukemia cell lines and is due to retroviral insertions in the 5' region of the gene in either the Evi-1 or the CB-1/FIM3 common sites of viral integrations. To examine the role of the Evi-1 gene in human leukemias we have cloned regions of the human locus corresponding to the coding region of the gene and regions corresponding to the Evi-1 and CB-1/FIM3 common sites of integrations. Using these probes we demonstrate that the human Evi-1 gene maps to chromosome 3q24-q28 in a region that is translocated in acute nonlymphocytic leukemias with a t(3;5)(q25;q34). By in situ hybridization with metaphase chromosomes from one patient with a 3;5 translocation, the Evi-1 gene was found to be translocated to the derivative 5 chromosome. However, no rearrangements were detected by Southern blot analysis with DNAs from three patients with a t(3;5) using probes from the Evi-1 or CB-1/FIM3 loci. No Evi-1 transcripts were detected with RNA from leukemic blasts of one patient with a t(3;5).

Blotting, Northern↗

PubMed1990

Unique expression of the human Evi-1 gene in an endometrial carcinoma cell line: sequence of cDNAs and structure of alternatively spliced transcripts.

Retroviral insertional activation of the expression of the Evi-1 is one of the most common events associated with transformation in murine myeloid leukemia. The murine Evi-1 gene encodes a 145 kDa nuclear, DNA binding protein that contains two domains containing seven and three sets of repeats of the zinc finger motif. During studies to determine the role of the Evi-1 gene in the transformation of human cells, we have found that the Evi-1 gene is uniquely expressed at low levels in HEC-1-A cells and at high levels in HEC-1-B cells, two related human endometrial carcinoma cell lines. cDNA clones were isolated and sequenced from the HEC-1-B cell line. The human gene is highly homologous to the murine gene and shows 91% and 94% homology in nucleotide or amino acid sequence respectively. In addition an alternatively spliced form of the gene was identified that encodes a protein with an internal deletion 315 amino acids including two of the zinc finger repeats. The possible basis for the unique expression of the Evi-1 gene by HEC-1 cells could not be determined by karyotype or Southern blot analysis.

Amino Acid Sequence↗

PubMed1990

Insertional mutagenesis and transformation of hematopoietic stem cells.

The mechanisms that are involved in the control of the normal differentiation of hematopoietic progenitor cells are largely unknown. Moreover, little is known concerning the types of genes that can alter the ability of hematopoietic progenitors to differentiate and cause transformation. One approach to the latter has been to use retroviral induced IL-3-dependent myeloid leukemia cell lines to identify transforming genes by their activation through insertional mutagenesis. This approach has implicated alterations in c-myb in transformation and has identified a novel transcriptional factor of the zinc finger family that is frequently activated in murine myeloid leukemias and in some cases of human AML. Using this approach it should be possible to identify additional myeloid transforming genes. The identification and characterization of the genes will provide important information and approaches to the study of the regulation of differentiation in normal hematopoiesis.

Acute Disease↗

PubMed1990

[A carcinosarcoma of the ovary: a case report and a review].

A carcinosarcoma of the ovary, which is a combined epithelial and mesenchymal tumor, is exceeding rare and usually has poor prognosis. Only four known cases, including this case, have been reported in Japan. In our case, after resection a combined chemotherapy consisting of cisplatinum, adriamycin cyclophosphamide and etoposide was administered and the patient had no uneventful postsurgical course.

Antineoplastic Combined Chemotherapy Protocols↗