Biomedical subjects

M Hanaoka

Publications and source records attributed to M Hanaoka.

At least 19 recordsLinked to original sources

PubMed1994

[Development of highly stereoselective reactions based on tricarbonyl(eta 6-o-trimethylsilylbenzaldehyde)-chromium(o) complex].

Treatment of the tricarbonyl(eta 6-o-trimethylsilylbenzaldehyde)chromium (O) complex with cyclic silyl enol ether species under the Mukaiyama condition resulted in highly stereoselective formation of syn-aldol products, whereas the aldol reaction with O-silyl ketene O,S-acetals afforded the corresponding anti-products in a highly stereoselective manner. The tricarbonyl(eta 6-o-trimethylsilyl-benzaldehyde)chromium(O) complex has also emerged as an excellent substrate for highly regio- and stereoselective construction of cis-3,5-isoxazolidine derivatives in the 1,3-dipolar cycloaddition reaction. By taking advantage of our newly developed aldol reaction, we have succeeded in highly stereocontroled syntheses of some biologically active compounds such as C-13 side chains of taxol and taxotère, the N-terminal amino acid of nikkomycin B and Bx, and antitumor styryllactones: goniofufurone, goniotriol, and goniothalenol.

Molecular Conformation↗

PubMed1994

[Hemothorax].

Explore the source record for details and available documents.

Hemothorax↗

PubMed1990

65-kilodalton protein phosphorylated by interleukin 2 stimulation bears two putative actin-binding sites and two calcium-binding sites.

We have previously characterized a 65-kilodalton protein (p65) as an interleukin 2 stimulated phosphoprotein in human T cells and showed that three endopeptide sequences of p65 are present in the sequence of l-plastin [Zu et al. (1990) Biochemistry 29, 1055-1062]. In this paper, we present the complete primary structure of p65 based on the cDNA isolated from a human T lymphocyte (KUT-2) cDNA library. Analysis of p65 sequences and the amino acid composition of cleaved p65 N-terminal peptide indicated that the deduced p65 amino acid sequence exactly coincides with that of l-plastin over the C-terminal 580 residues [Lin et al. (1988) Mol. Cell. Biol. 8, 4659-4668] and has a 57-residue extension at the N-terminus to l-plastin. Computer-assisted structural analysis revealed that p65 is a multidomain molecule involving at least three intriguing functional domains: two putative calcium-binding sites along the N-terminal 80 amino acid residues; a putative calmodulin-binding site following the calcium-binding region; and two tandem repeats of putative actin-binding domains in its middle and C-terminal parts, each containing approximately 240 amino acid residues. These results suggest that p65 belongs to actin-binding proteins.

Actins↗

PubMed1990

Efficient tumor suppression by glioma-specific murine cytotoxic T lymphocytes transfected with interferon-gamma gene.

We have recently shown that exogenous expression of the mouse interferon-gamma (IFN-gamma) gene augmented the cell-killing potential of a line of cytotoxic T lymphocytes (CTLs) specific against a murine glioma line (203-glioma). In the present work, we further investigated the in vivo antitumor effects of the E gamma-6 and E gamma-9 sublines of this CTL line transfected with the IFN-gamma gene. Using the Winn assay to test the neutralization of subcutaneous gliomas, we determined that these CTL sublines were more effective than the E-4 parent CTL line and that suppression of the tumor growth was dependent on the number of effector cells (CTLs). Moreover, intravenous injection of E gamma-9 cells was more effective in suppressing the tumor growth than intravenous injection of E-4 cells. These results suggest that transfection of antitumor effector cells with the IFN-gamma gene could improve the efficacy of adoptive immunotherapy against cancer.

Animals↗

PubMed1990

Characterization of interleukin 2 stimulated 65-kilodalton phosphoprotein in human T cells.

We have characterized the cellular proteins which are rapidly phosphorylated by interleukin 2 (IL 2) in a human IL 2 dependent cell line. When treated with IL 2, the phosphorylation of five proteins, 65, 50, 37, 24, and 21 kDa, was found in IL 2 dependent cell lines by two-dimensional gel electrophoretic analysis. After cell conversion from an IL 2 dependent state to an IL 2 independent state, one of the five phosphoproteins, the 65-kDa protein, became constitutively phosphorylated even without addition of IL 2. Also, in other IL 2 independent cell lines, such as KUT-2 and HUT-102, constitutive phosphorylation of the 65-kDa protein occurred without IL 2-stimulation. So our researchers were focused on biochemical characterization of the 65-kDa protein. It was found that the 65-kDa protein was one of the major cellular proteins by comparing the results of two-dimensional gel electrophoretic analysis of [32P]Pi-labeled and [3H]leucine-labeled cellular proteins and peptide mapping analysis. Subcellular fractionation studies indicated that the 65-kDa protein is a cytosol protein. The 65-kDa protein was purified from cytosol of a human T cell line, and its amino acid composition and amino acid sequences of its three oligopeptides were determined. It was found that the 65-kDa protein is identical with 1-plastin.

Amino Acid Sequence↗

PubMed1990

[Work load of nursery teachers in a nursery school. Relationship between age of children and work load].

Time study of six nursery teachers of children aged 0 to 5 was conducted at a public nursery school to ascertain their work load, and the results obtained were examined from the following two viewpoints. First, comparison was made between the results obtained by the following two methods of analysis; 1) An observer recorded the work contents and the working postures of a nursery teacher at place of work in one-minute units. 2) The nursery teacher was recorded on video tape and the analysis of the work contents and the working postures was made in one-second units. Second, comparison was made between the work loads (work content, working posture, walking, lifting children and recess) of nursery teachers of children of different age group. The results could be summarized as follows: 1. The results of analysis made by the two methods of the work contents and the working postures were in good agreement. 2. The higher the age of children in the class, the longer was the cumulative time for "indoor group nurture" and the shorter was the cumulative time for "help and care for feeding", "help and care for afternoon nap" and "help and care for excretion". 3. The higher the age of children in the class, the longer was the cumulative time of "standing" in a nursery teacher and the shorter was the cumulative time of "sitting on the floor". The total time of "bending forward", "squatting" and "rising on the knee" occupied 20% of the working hours on an average and was not related clearly to the age of children. 4. The frequency of walking was not found to be related to the age of children. 5. The nursery teachers of children aged 0 and 1 lifted children much more frequently among all nursery teachers. 6. The recess away from children was rarely observed throughout the day.

Age Factors↗

PubMed1989

Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor.

The sequence of 5,037 amino acids composing the ryanodine receptor from rabbit skeletal muscle sarcoplasmic reticulum has been deduced by cloning and sequencing the complementary DNA. The predicted structure suggests that the calcium release channel activity resides in the C-terminal region of the receptor molecule, whereas the remaining portion constitutes the 'foot' structure spanning the junctional gap between the sarcoplasmic reticulum and the transverse tubule.

Amino Acid Sequence↗

PubMed1989

Expression of proto-oncogene products during drug-induced differentiation of a neuroblastoma cell line SK-N-DZ.

Four human neuroblastoma cell lines exhibited differences in their ability to differentiate into neuron-like cells in response to three different treatments, serum deprivation, or additions of dibutyryl cyclic-AMP or retinoic acid. Expression of N-myc gene product was reduced in neuroblastoma cell line SK-N-DZ differentiated by retinoic acid as compared with untreated cells. On the contrary, expression of c-src gene product, pp60c-src, was considerably enhanced in differentiated SK-N-DZ cells. Tyrosine phosphorylation of several cellular proteins was found to be enhanced in differentiated cells. Alteration in expression of these proto-oncogene products might be important in the differentiation of neuroblastoma cells into neuron-like cells.

Bucladesine↗

PubMed1989

Detection of fos oncogene products by monoclonal antibody FO-120 in lymphoproliferative disorders.

We investigated the expression of fos oncogene proteins in lymphoproliferative disorders, using a monoclonal antibody (FO-120) that was prepared against a synthetic oligopeptide of fos protein (amino acid sequence from 127 to 152). Although peripheral blood leukocytes were rarely positive for FO-120, they were transiently stained after lectin (PHA) stimulation. After culture with IL-2 for 1 or 2 weeks, less than 40% of the lymphocytes weakly reacted with FO-120, whereas strongly positive cells were detected in more than 70% of cells in half the T-cell lines established from preleukemic state of adult T-cell leukemia (pre-ATL) and all of ATL derived T-cell lines. All in vivo specimens of non-Hodgkin's malignant lymphomas, except for one case of T-cell lymphoma were also strongly positive. In addition, the extent of the antibody reactivity correlated with the histopathological grade of malignancy in B-cell lymphoma. The reactivity to most AILD-IBL lesions overlapped with that to T-lymphomas, and could be distinguished from that to reactive lesions. FO-120 appears to be a useful tool for detecting early neoplastic changes in lymphoproliferative disorders.

Antibodies, Monoclonal↗

PubMed1989

Murine intracerebral interleukin-2 injection: pathological and immunological effects.

The authors have investigated whether specific pathological changes and antibodies against interleukin-2 (IL-2) are induced after intracerebral administration of recombinant IL-2 (rIL-2). In addition, IL-2 receptor (IL-2R) expression was checked on the cell surface of normal brain tissues before and after the intracerebral infusion. Reconstituted rIL-2 (specific activity 1.2 x 10(7) U/mg protein) was injected into the right cerebral hemisphere of normal adult C57BL/6 mice in three different dose groups, each receiving single or multiple infusions of 8, 32, or 80 U. In sham control experiments, mouse albumin purified by gel filtration and ion exchange chromatography and adjusted to the same concentration of protein as rIL-2 was injected into mice at various doses. Anti-IL-2 antibodies were measured by an enzyme-linked immunosorbent assay concurrently with assessment of IL-2 activity in serum. The IL-2R expression was determined by using immunofluorescence techniques with monoclonal antibodies against mouse IL-2R. Since histological alteration after rIL-2 injection did not differ from that in the sham control preparations, it seems that there is no direct toxic action of rIL-2 on normal brain tissues. Interleukin-2 antibodies were produced at low levels only in mice injected repeatedly at the maximum dose, and levels were insignificant in other groups. Serum levels of IL-2 activity remained low. The IL-2R expression within the brain was not enhanced within 8 weeks following the intracerebral administration of rIL-2, suggesting that direct intracerebral infusion of rIL-2 may be safely used in the immunotherapy of brain tumors.

Animals↗

PubMed1989

Acute-phase response of angiotensinogen in rat adjuvant arthritis.

The plasma level of angiotensinogen during the chronic phase of inflammation was studied for comparison with those of other acute-phase reactants in rat adjuvant arthritis. In response to a single injection of Freund's complete adjuvant, this level exhibited a transient increase during the first 24 h. By contrast, increased levels of plasma T-kininogen and alpha 2-macroglobulin, typical acute-phase reactions in the rat, were maintained during the 4-week experimental period. These results suggest that the hepatic synthesis of angiotensinogen is stimulated only in the early phase of chronic inflammation, and therefore that the mechanism underlying the acute-phase response of angiotensinogen is distinct from those currently suggested for other acute-phase reactants.

Acute-Phase Reaction↗

PubMed1988

Augmentation of tumor targeting in a line of glioma-specific mouse cytotoxic T-lymphocytes by retroviral expression of mouse gamma-interferon complementary DNA.

As an initial approach to experiments directed toward effective adoptive immunotherapy for cancer using lymphokine genes, we transferred retrovirally a complementary DNA encoding mouse gamma-interferon (IFN-gamma) into a specific cytotoxic T-lymphocyte clone, designated E-4, against 203 glioma cells (a 20-methylcholanthrene-induced mouse glioma line) and confirmed the efficacy of IFN-gamma production from the exogenous gene on augmentation of tumor targeting. Of five, two gene-transferred subclones constitutively produced 8 to 10 times the amount of IFN-gamma as compared with the parental E-4. Correspondingly, these two subclones exhibited 2 to 3 times higher killing activity against 203 glioma than the parental cells; the enhancement of the killing activities was abrogated by an adequate addition of anti-IFN-gamma antibody. No alteration was seen after the gene transfer in cell surface phenotypes, Thy-1+, Lyt-1-, Lyt-2+,3+, and asialo-GM1-. The surface expression of a major histocompatibility complex Class I antigen, H-2Kb, was not altered remarkably, but the Class II antigen, I-Ab, was partially and slightly enhanced on the two IFN-gamma-producing sublines mentioned above on fluorescence-activated cell sorter analysis. Since it is considered that in the vicinity of the constitutively IFN-gamma producing cytotoxic T-lymphocyte cells tumor cells are exposed to a high concentration of IFN-gamma, the cells may be stimulated to induce or enhance the expression of surface antigens including major histocompatibility complex antigens as well as tumor-associated antigens relevant to immune recognition. The 203 glioma cells pretreated with IFN-gamma were more efficiently killed by both the parental E-4 and the gene-transferred sublines. Taken together, the results suggested that the augmented specific tumor-killing activity of our gene-transferred cytotoxic T-lymphocytes was ascribed to the constitutive production of IFN-gamma derived from the exogenous gene.

Animals↗

PubMed1988

Immunoregulatory effects of interleukin 2 and interferon on syngeneic murine malignant glioma-specific cytotoxic T-lymphocytes.

The effects of interleukin 2 (IL2) and interferon (IFN) on the generation and lytic activation of syngeneic murine malignant glioma (a methylcholanthrene-induced ependymoblastoma of C57BL/6 mouse origin, 203-glioma)-specific cytotoxic T-lymphocyte (G-CTL) were investigated. The surface marker analysis showed that G-CTLs from both intracranial and s.c. tumor-bearing mice were composed of thymectomy-resistant (mature) Lyt-1-.2.3+ and thymectomy-sensitive (immature) Lyt-1+.2.3+ CTLs, which markedly decreased concurrently with increased intracranial pressure. G-CTLs were confirmed to be activated with target specificity by both factors in a different way. The CTL activation by IL2 (20 units/ml) remained for a longer time, although a lag time of 5 days after initial culture was required. IL2 influenced Lyt-1+.2.3+ CTLs to proliferate and develop the lytic potential. In contrast, even a 3-h incubation with IFN (1000 units/ml) could enhance the cytotoxicity, but the augmenting effects were observed no longer than 5 days later. IFN activated Lyt-1-.2.3+ CTLs and increased their proportion of the total cell population with a simultaneous decrease of Lyt-1+.2.3+ CTLs. Therefore, it was suggested that IL2 may provide a growth of CTL populations and that IFN can accelerate recruitment of new effectors, causing activation of the lytic process.

Animals↗

PubMed1988

Establishment of a hemopoietic stimulating factor producing murine leukemia cell lines: pathogenesis of granulocytosis in L8313 bearing mice.

Thy 1.2+ cells of L8313 leukemia bearing mice were previously shown to produce granulocyte macrophage colony stimulating activity (GM-CSA) and interleukin-3 (IL-3). Cell lines with the Thy 1.2+ phenotype were established from spleen cells of the leukemic mice, and were designated STIL-3 C5 and STIL-3 D10. They produced and released GM-CSA and IL-3 activities in culture supernatant. C5 cells were phenotypically Thy 1.2+ and Lyt 2.1+ with rearrangement of the T-cell receptor beta chain gene also being identified. D10 was Thy 1.2+ and L3T4+ with T-cell receptor beta chain gene rearrangement not detectable. Since the same sized rearranged bands were observed between C5 and L8313 leukemic mouse spleen cells, it is indicated that C5 cells are derived from the leukemic cells. Inoculation of these cells into mice induced granulocytosis. These results indicated that L8313, which was thought to be a "granulocytic leukemia", is actually a T-cell leukemia, whose granulocytosis is a leukemoid reaction induced by normal hemopoietic cells responding to the hemopoietic stimulating factors, GM-CSA and IL-3, produced by the leukemic T cells. Thus, L8313 should be known as a T-cell leukemia associated with granulocytosis.

Animals↗

PubMed1988

Letterer-Siwe disease: immunopathologic study with a new monoclonal antibody.

Three cases of Letterer-Siwe disease were studied with the monoclonal antibody Lag, which reacts to the antigen on the membranes of Birbeck granules and related structures of human Langerhans cells. Both lymph nodes and lesional skin contained abundant Lag-positive cells. By two-dimensional gel electrophoresis, antigenic substances in the lymph nodes of patients with Letterer-Siwe disease were found to have the same molecular weight of 40,000 dalton and isoelectric points extending from 4.7 to 6.5 as those in normal human skin and lymph nodes. Our results support the contention that Letterer-Siwe disease is a proliferative disorder of Langerhans cells. A double-staining method with Lag and anti-T6 antibody revealed that Lag reacted to 70% of T6-positive cells in the lymph nodes but to almost all such cells in skin lesions of patients with Letterer-Siwe disease, suggesting that the proliferating cells consist of at least Lag+, T6+, and Lag-, T6+ subpopulations.

Antibodies, Monoclonal↗

PubMed1988

Lethal graft-versus-host disease in nude mice. I. Establishment of model systems.

We examined whether nude mice, which are deficient in T cell function, could be used as a model for induction of lethal graft-versus-host disease. Nude mice injected with MHC-disparate spleen cells exhibited only transient GVH reaction such as splenomegaly. Inoculation of B6 spleen cells into BALB/c nude mice produced high titers of alloantibodies to the donor cells. These alloantibodies eliminated host-MHC-reactive donor T cells from the host. After abolition by 400 rads irradiation of the capacity of nude mice to produce antibody, lethal GVHD could be induced by allogeneic spleen cell transfer and was mediated by donor T cells. This lethal GVHD was prevented by prior administration of antidonor alloantibody to the irradiated recipients at least 24 hr before donor-cell grafting. The role of alloantibody was substantiated in 2 other combinations in which little or no alloantibodies to donor spleen cells were produced. Engraftment of either MHC-identical but non-MHC disparate donor spleen cells into BALB/c nude mice or of parental spleen cells into F1 nude mice resulted in death mediated by T cells. In addition, irradiated BALB/c nude mice inoculated with non-MHC-incompatible B10.D2 spleen cells were much more sensitive to alloaggression by the donor cells than were nonirradiated hosts, indicating the presence of some radiation-sensitive component(s) acting in nude mice against GVHD induction by donor T cells. Thus the nude mouse is considered to be a useful recipient for clarifying the basic mechanisms involved in lethal GVHD.

Animals↗

PubMed1988

Persistent and enhanced expression of c-fos gene products in various kinds of human hematopoietic cell lines. Immunofluorescence study using prepared monoclonal antibodies.

Two kinds of monoclonal antibodies recognizing fos proto-oncogene (c-fos) products were prepared using a synthetic oligopeptide corresponding to amino acids 127-152 of the fos oncogene products. These monoclonal antibodies (FO-120 & FO-145) detected fos gene products induced in a human monocyte cell line (U-937) by phorbol acetate (TPA) and induced in both human and mouse fibroblast cell lines (284, BALB/c 3T3) by serum-stimulation. One of the monoclonal antibodies (FO-120) reacted with 50-kDa and 42-kDa proteins and the other antibody (FO-145) reacted with a 30-33-kDa protein. The expression of the fos gene in various human hematopoietic cell lines was investigated using these prepared monoclonal antibodies. While almost all hematopoietic cell lines tested reacted with these monoclonal antibodies to various degrees, the majority of normal peripheral blood lymphocytes cultured with lectin (PHA) and interleukin 2 (IL-2) did not, suggesting that cells of some permanent hematopoietic cell lines, irrespective of their lineage specificity and growth factor dependency, continuously express the fos oncogene. These monoclonal antibodies may be useful for detecting early neoplastic changes in hematopoietic cells.

Animals↗

PubMed1988

Simultaneous measurement of transferrin receptor and DNA content of human IL 2 dependent T cells by flow cytometry.

This paper describes a method which enables the simultaneous measurement of both the concentration of cell surface receptors and the DNA content of individual lymphoid cells. Cells fixed with PLP (periodate-lysine-paraformaldehyde) were treated with ribonuclease (RNase). Transferrin receptors were then successively bound with monoclonal antibody against them and FITC-labeled antibody against the monoclonal antibody. Cells thus treated were stained with propidium iodide and two-parameter flow cytometric analysis was carried out. Using this method, the expression of transferrin receptors on lymphoid cells was analyzed in relation to the action of T-cell growth factor (IL 2). It was found that cells in the G1 phase were stimulated by IL 2 which increased transferrin receptor concentration after a lag of a few hours. Subsequently, the cells entered the S phase and the receptor levels remained high throughout the S, G2 and M phases of the cell cycle.

Cell Cycle↗