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Biomedical subjects

K Motoyoshi

Publications and source records attributed to K Motoyoshi.

At least 199 records · Page 11Linked to original sources
PubMed1986

Discrepancy between the production of interleukin 1, alpha-interferon and granulocytic colony-stimulating activity of human monocytes stimulated by a partially purified human urinary colony-stimulating factor.

A partially purified human urinary colony-stimulating factor (pp-CSF), with a specific activity of 1.0 X 10(6) U/mg protein, was purified by using a stepwise DEAE-cellulose anion exchange chromatography and a molecular sieve high performance liquid chromatography (TSK gel G3000SW) sequentially. Production of three kinds of monokines by human peripheral blood monocytes was tested in vitro, which contained granulocytic colony-stimulating activity (G-CSA), interleukin 1 (IL-1) and interferon alpha (IFN-alpha). Human peripheral blood mononuclear cells were separated by Ficoll-Hypaque density gradient centrifugation, and monocytes were obtained by adherence to dishes. Although pp-CSF stimulated monocytes to produce G-CSA in vitro, it failed to stimulate the IL-1 or IFN-alpha production. The discrepancy between production or release of these three kinds of monokines by human monocytes stimulated with pp-CSF suggests that pp-CSF preferentially stimulates human monocytes to produce G-CSA. To test the effects of pp-CSF on human monocytes in vivo, we performed i.v. infusions of pp-CSF to four volunteers, and we then took peripheral blood monocytes. Compared to before the infusion of pp-CSF, G-CSA production by monocytes was enhanced, while production of IL-1 and IFN-alpha was not enhanced after the infusion of pp-CSF. These results suggest that pp-CSF stimulates monocytes to produce G-CSA but not to produce IL-1 nor IFN-alpha.

Cells, Cultured↗
PubMed1986

Clinical application of partially purified human urinary colony-stimulating factor.

Human urinary colony-stimulating factor (CSF-HU) has been partially purified using five purification procedures (preparation II) for clinical use. In phase I study, six healthy volunteers were injected with preparation II of CSF-HU by a single intravenous droplet infusion. Two volunteers received 4 X 10(6) units (U), two received 8 X 10(6) U, and the final two received 1.2 X 10(7) U of preparation II of CSF-HU. Five volunteers did not have any symptoms, but one volunteer who received 1.2 X 10(7) U complained of transient, mild chilliness during the infusion. Liver function test, coagulation test, urine analysis and patterns of electrocardiogram did not change after infusions. In phase II study on preparation II of CSF-HU, forty-four patients with urological malignancy were infused with a daily dose of 8 X 10(6) U CSF-HU for 7 days from the next day after the end of the second courses of two consecutive courses of the same chemotherapeutic regimen. The average nadirs of leukocytes and granulocytes of 33 evaluable cases were higher in CSF-HU-infused courses (second courses) than in control courses (first courses), with statistic significances. The average period under 2,000 leukocytes/mm3 and the average period under 500 granulocytes/mm3 were shorter in CSF-HU-infused courses than in control courses. These results might indicate that infusions of preparation II partially protect the patients from granulocytopenia after anticancer chemotherapies.

Adult↗
PubMed1986

Mode of action of human urinary colony-stimulating factor.

Human urinary colony-stimulating factor (CSF-HU) has been highly purified using procedures containing DEAE cellulose, phenyl Sepharose CL-4B, Sephadex G-200, hydroxylapatite, and high performance liquid chromatography. The final preparation had a specific activity of 3.3 X 10(7) U/mg protein. Although the purified CSF-HU was not active on human monocyte-depleted bone marrow cells, it stimulated human peripheral blood monocytes obtained from five healthy volunteers to produce human active granulocytic colony-stimulating factor (G-CSF), which stimulated human monocyte-depleted bone marrow cells to form granulocytic colonies. The human G-CSF-producing activity of CSF-HU was not neutralized by polymyxin B, which is known to inhibit the effect of endotoxin. Newly produced G-CSF had an approximate molecular weight of 24,000 daltons as judged by chromatography on Sephadex G-150. These results indicate that CSF-HU stimulates human monocytes to produce human G-CSF in vitro.

Colony-Stimulating Factors↗
PubMed1986

Protective effect of partially purified human urinary colony-stimulating factor on granulocytopenia after antitumor chemotherapy.

We conducted a randomized crossover study comparing the hemopoietic effect of partially purified human urinary colony-stimulating factor (CSF-HU, an active drug) and human serum albumin (HSA, a control drug) in 24 patients with malignant lymphoma, solid tumors, or multiple myeloma who were receiving two consecutive courses of the same chemotherapeutic regimen. Patients received daily 2-4 X 10(6) units of CSF-HU or an equal amount of protein HSA for five days after the end of the courses of chemotherapy. Assignment to CSF-HU or HSA was determined by the envelope method. The average number of blood granulocytes of 24 cases on day 7 after chemotherapy was 2116 +/- 1649 in CSF-HU-infused courses, which was significantly higher than in HSA-infused courses (1520 +/- 1022) (p less than 0.05). The average time that patients had fewer than 2000 granulocytes/mm3 was 7.6 +/- 4.4 days in CSF-HU-infused courses and 10.3 +/- 5.0 days in HSA-infused courses (p less than 0.02). Fever greater than 38 degrees C was the most frequent side effect, occurring in 32% of the patients receiving CSF-HU infusions. A reduction in the neutropenic interval in CSF-HU-infused courses was observed in patients with fever, as well as in those without fever. Infusions of CSF-HU did not change the number of other hematological parameters, such as erythrocytes, platelets, monocytes, and lymphocytes. These results suggest that CSF-HU infusions may partially protect the patients from granulocytopenia after anticancer chemotherapy.

Agranulocytosis↗
PubMed1985

Sebaceous gland hyperplasia on rabbit pinna induced by tetradecane.

To investigate the pathologic changes of sebaceous glands during comedo formation induced by topically applied substances in a rabbit pinna model, purified tetradecane was inuncted on the ventral aspect of the rabbit pinnas once a day for a week. Histologically, a marked hyperplasia of sebaceous glands, epidermis, and follicular epithelium was seen. These remarkably enlarged sebaceous glands were examined histochemically and ultrastructurally. The acinus size and cell population of the hyperplastic sebaceous gland were significantly increased over those of the normal sebaceous gland. By N-(7-dimethylamino-4-methyl-3-coumarinyl)maleimide staining, normal distribution patterns of sulfhydryl (SH) and disulfide (SS) were seen in the peripheral to differentiating layers in the hyperplastic sebaceous glands. In the terminally differentiated layer, the brilliant SH fluorescence was gradually decreased and the SS fluorescence was gradually increased in intensity, indicating that most SH groups in the sebaceous cells were converted to SS linkages before holocrine secretion. By transmission electron microscopy, several cell layers of undifferentiated sebaceous cells were observed at the periphery of the large sebaceous gland. The differentiating sebaceous cells produced a large number of lipid droplets, which were produced in either rough or smooth endoplasmic reticulum. These cells were abruptly converted into homogeneously electron-dense cells which formed several layers. These homogeneous cells gradually lost their electron density before holocrine secretion. These findings indicate that the sebaceous cells in the hyperplastic sebaceous glands undergo a magnified step-by-step cell differentiation and play a role in slightly modified lipid formation, and that there may be an increased production of sebum in the rabbit pinna model. This is the first report of sebaceous hyperplasia induced by a topically applied substance on skin surface, except for androgens. The hyperplastic sebaceous glands could serve as a model for investigations of sebaceous cell differentiation and lipid formation.

Administration, Topical↗
PubMed1985

Interleukin 1 (IL 1)-dependent lymphokine production by human leukemic T cell line HSB.2 subclones.

Cloning of a human T cell leukemic cell line, HSB.2, was performed by a limiting dilution method to obtain clones with high levels of IL 2 production. None of the subclones that were obtained produced IL 2 constitutively, and only a low level of IL 2 was produced by the stimulation of these subclones with phytohemagglutinin (PHA) alone. High levels of IL 2 production (greater than 300 U/ml) were observed in several clones when stimulated with a cocktail of PHA and IL 1. Among them, HSB.2-A7-D2, A7-D9, or C5-B2 subclones, which were selected after cloning twice, were most effective in IL 1-dependent IL 2 production. HSB.2 subclones exhibited IL 1-dependent production of a variety of lymphokines other than IL 2, e.g., interferon-gamma (IFN-gamma), B cell growth factor (BCGF), and colony-stimulating factor (CSF). We observed that subclones with high IL 2-producing capacity tended to produce high levels of IFN-gamma or BCGF as well, while the capacity of CSF production was not parallel to these properties. Although several subclones were found to produce IFN-gamma and BCGF simultaneously with minimal IL 2 activity, no subclones with an exclusive BCGF production were obtained. Furthermore, when supernatants from the stimulated A7-D9 subclone were applied to an Ultro-gel AcA54 gel chromatography, it was revealed that IL 2 activity (m.w. 17K to 18K) and IFN-gamma (40K to 45K) were clearly separated, whereas two peaks of BCGF activity coincided with each peak of IL 2 and IFN-gamma, respectively. On the other hand, CSF activity was eluted at a different peak (30K to 35K). These data indicate that IL 2, IFN-gamma, and CSF activities are based on distinct molecules, whereas BCGF activities are indistinguishable from IL 2 and IFN-gamma. The HSB.2 subclones thus selected will provide a useful model for delineating the mechanism of IL 1-dependent lymphokine(s) production, and are a promising candidate for better lymphokine(s) producers.

Cell Line↗
PubMed1985

Human colony-stimulating activity-producing tumor: production of very low mouse-active colony-stimulating activity and induction of marked granulocytosis in mice.

We reported previously a human lung cancer which induced marked granulocytosis both in the patient and in the tumor-transplanted nude mice (G-1 mice), and whose conditioned media (G-1-T-CM) contained both human-active colony-stimulating activity (h-CSA) (810 colonies/ml) and mouse-active colony-stimulating activity (m-CSA) (530 colonies/ml), suggesting that the CSA produced by the tumor caused granulocytosis both in the patient and in G-1-mice. We reported here another human lung cancer which induced marked granulocytosis both in the patient and in the tumor-transplanted nude mice (G-2-mice). However, in contrast to the tumor reported by us previously, the conditioned media of this tumor (G-2-T-CM) contained only very weak m-CSA (86 colonies/ml), although h-CSA was very strong (7332 colonies/ml). The ratio of m-CSA to h-CSA in G-1-T-CM (0.654) was 55-fold higher than that in G-2-T-CM (0.012). Gel filtration of G-2-T-CM on a Sephadex G-150 column denied the presence of colony-inhibiting activity which inhibited m-CSA in G-2-T-CM. Since the addition of serum obtained from G-2 mice did not enhance m-CSA in G-2-T-CM, it seems unlikely that the G-2 tumor produced an inactive form of m-CSA which was activated in G-2 mouse serum. G-2-T-CM tended to maintain the number of colony-forming units in spleen in mouse bone marrow during liquid cultures, while G-1-T-CM did not. These results may indicate that the G-2 tumor produced a humoral factor which has very weak CSA and acts on stem cells rather than on committed progenitors.

Animals↗
PubMed1985

Two antigenically different types of colony-stimulating activities in sera of patients with aplastic anemia.

Sera obtained from seven normal volunteers and 11 patients with aplastic anemia were assayed for two types of colony-stimulating activity (CSA) using monolayer agar cultures containing human unfractionated and monocyte-depleted bone marrow cells. Sera obtained from normal volunteers had low CSA for unfractionated bone marrow cells and no CSA for monocyte-depleted bone marrow cells. On the other hand, sera obtained from patients with aplastic anemia (patients' sera) had moderate CSA for both unfractionated and monocyte-depleted bone marrow cells. The patients' serum CSA titer for unfractionated bone marrow cells rose markedly with the addition of a small amount of diluted control rabbit serum (control serum) into the CSA assay system, while it did not rise with addition of the same amount of diluted rabbit antiserum against partially purified human urinary colony-stimulating factor (CSF) (antiserum). The patients' serum CSA titer for monocyte-depleted human bone marrow cells rose with the addition of control serum as well as antiserum. These findings indicate that the addition of rabbit serum enhances colony formation by human bone marrow cells in the presence of aplastic anemia sera as a source of CSA, and that sera of patients with aplastic anemia contain two antigenically different types of CSA: one that is active on human unfractionated bone marrow cells and partially neutralized by the addition of antiserum, and another that is active on human monocyte-depleted bone marrow cells and is not neutralized by the addition of antiserum.

Adult↗
PubMed1984

New findings on the proteins of sebaceous glands.

In order to understand the distribution and concentration of proteins with -SH groups or S-S linkages in sebaceous cells during differentiation and holocrine secretion of sebaceous glands, skin specimens from the inner side of ears of New Zealand white rabbits were examined histochemically and ultrastructurally. DACM (N-[7-dimethylamino-4-methyl-3-coumarinyl] maleimide) staining method showed that proteins containing -SH groups were present in the cells (cytoplasm and nuclei) in all layers from the peripheral to the terminally differentiated cells of sebaceous glands and that proteins containing S-S linkages were present in the terminally differentiated cells and their pyknotic nuclei but not in the peripheral and differentiating cells of sebaceous glands. Lipid droplets in all sebaceous cells contained neither -SH groups nor S-S linkages. Ultrastructurally, the terminally differentiated cells were very electron dense and seemed to be abruptly formed from the differentiating cells that were producing lipid droplets. These findings indicate that the conversion of -SH groups to S-S linkages of proteins also occurs in sebaceous glands as in epidermis and hair.

Animals↗
PubMed1984

Characteristic expression of glycosphingolipid profiles in the bipotential cell differentiation of human promyelocytic leukemia cell line HL-60.

Changes of glycosphingolipids (GSLs) in the bipotential cell differentiation of human promyelocytic leukemia cell line HL-60 cells were investigated by high-performance thin-layer chromatography (HPTLC), with special reference to morphological and functional changes, such as phagocytosis and nitroblue tetrazolium (NBT) reduction. Nine molecular species of neutral GSLs and 13 or more species of sialo-GSLs, ie, gangliosides, were detected on the HPTLC chromatograms for untreated HL-60 cells. The major components were ceramide dihexoside (CDH), GM3, and sialo-paragloboside (SPG). When HL-60 cells were induced to differentiate into both myeloid mature cells and macrophage-like cells in vitro, no new molecular species of GSLs specific for one of the cell differentiations was induced, but distinctive quantitative changes in the GSL composition were definitely observed between the two cell differentiations. During the myeloid differentiation induced by either dimethylsulfoxide (DMSO) or retinoic acid (RA), CDH, paragloboside (PG), and gangliosides having longer sugar moieties characteristically increased with a concomitant decrease of GSLs with shorter sugar chains, such as ceramide monohexoside (CMH) and GM3, and the GSL composition profile of myeloid differentiation-induced HL-60 cells became more similar to that of normal human granulocytes. However, some marked differences were noted between the induced HL-60 cells and the normal granulocytes, especially in the ganglioside compositions. These differences might reflect either some deficiency in the in vitro myeloid differentiation or some leukemic properties of HL-60 cells. In marked contrast to the change of GSL composition during myeloid differentiation, a remarkable increase of GM3, with a concurrent marked decrease of CDH, was observed in the process of cell differentiation into macrophage-like cells with 12-O-tetradecanoyl-phorbol-13-acetate (TPA), which suggested an increase in the biosynthesis of GM3. These results demonstrate that HL-60 cells express distinct GSL profiles, depending not only on maturation stages but also on differentiation directions.

Cell Line↗