PubMed Health⌕ Search

Biomedical subjects

M Bessho

Publications and source records attributed to M Bessho.

At least 91 records · Page 5Linked to original sources
PubMed1990

Terminal differentiation to mature neutrophils and eosinophils in suspension culture of the blast progenitors in acute myeloblastic leukemia.

The blasts obtained from three freshly diagnosed acute myeloblastic leukemia (AML) patients were cultured in suspension to determine whether leukemic blast progenitors can indeed differentiate to form mature granulocytes. One patient was AML M2. The other two patients were bilineal and biphenotypic leukemia, respectively. Media conditioned by human bladder carcinoma line 5637 (5637-CM) or recombinant human granulocyte colony-stimulating factor (rhG-CSF) was added to stimulate growth. In suspension, clonogenic cells grew for 1-3 weeks in two patients, while they did not increase in one patient. After repeated subculture, cells of blast morphology decreased in percentage and polymorphonuclear neutrophils, eosinophils, and monocyte-macrophages appeared. Lymphoid cell component of the patient 2, who was diagnosed as bilineal leukemia by dual-color immunofluorescence analysis, decreased in number after suspension culture and cells of myeloid phenotype became dominant. The findings show that clonogenic blast progenitors can renew themselves and can also undergo terminal differentiation to mature end cells.

Adult↗
PubMed1990

[Aspergillosis of the maxillary sinus in a patient with Ph1 positive acute lymphoblastic leukemia: a case report].

A 46-year-old woman was admitted to our hospital because of leukocytosis. A diagnosis of acute lymphoblastic leukemia (FAB: L2 type) was made by reviewing peripheral blood smear and bone marrow aspirate. Chromosome analysis showed the presence of Philadelphia chromosome. A combination chemotherapy with L-asparaginase, doxorubicin, vincristine, and prednisolone was started, but complete remission was not achieved. During a neutropenic period after combination chemotherapy with doxorubicin, vincristine, vinblastine, and VP-16, high fever and tender swelling of the right cheek were noticed. A diagnosis of maxillary sinusitis was made with tomography and CT scan of the maxillary sinus. Since culture of the aspirate from the maxillary sinus grew aspergillus, a diagnosis of aspergillosis of the maxillary sinus was made. Immediately after the intravenous administration of amphotericin B and the lavage of the sinus with amphotericin B was started, high fever subsided and clinical improvement was observed. Several regimens of chemotherapy failed to obtain hematological remission, she died of sepsis of Enterobactor cloacae without evidence or relapse of dissemination of aspergillosis after initial successful treatment. While a few cases with aspergillus maxillary sinusitis were reported in leukemic patients, the possible occurrence of this complication must be kept in mind in a severe neutropenic period after intensive chemotherapy. The combination of intravenous administration and local lavage of amphotericin B appeared to be an effective treatment in the Aspergillus maxillary sinusitis.

Amphotericin B↗
PubMed1989

NAD and NADH values in rapidly sampled dog heart tissues by two different extraction methods.

To clarify the most quantitative extraction method for the determination of NAD and NADH in dog heart tissues, both pyridine dinucleotides were extracted from normal and ischemic heart tissues by the Klingenberg method and the Karp method and determined by bacterial luciferase. Tissues from normal beating hearts were sampled by a specially developed freeze-clamping device in 120 ms to minimize ischemic NADH production during sampling. Samples were obtained from both the subendocardium and the subepicardium of the frozen heart tissues. In the Klingenberg method, NAD and NADH were separately extracted with 0.6 M HClO4 and 0.5 M KOH in 50% ethanol, respectively. Both pyridine dinucleotides were simultaneously extracted with 70% ethanol in 0.01 M phosphate buffer in the Karp method. The mean values of NAD and NADH in the normal tissues were 5.08 +/- 0.84 and 0.18 +/- 0.10 nmol/mg protein, respectively, with a NAD/NADH ratio of 25-30 by the Klingenberg method. While the values by the Karp method were 4.37 +/- 0.68 and 0.09 +/- 0.04 nmol/mg protein, with a NAD/NADH ratio of 55-65. The efficiency of extraction of both pyridine dinucleotides by the Karp method was lower than that by the Klingenberg method in all tested samples and states of the tissues. These results suggest that the Klingenberg method is preferable for the extraction of both pyridine dinucleotides from dog heart tissues and that the mean NAD/NADH ratio in normal dog heart tissues is 25-30.

Animals↗
PubMed1989

A rapid cross-sectioning and freeze-clamping device for the beating canine heart.

A new sampling method of cross-sectioning the canine heart in situ was developed. A mechanical device, driven by spring power, enabled cross-sectioning of a short-axis plane of the beating canine heart (4 mm thick) with high speed rotating blades, at a pre-determined phase of the cardiac cycle, and instantaneous freeze-clamping (2.4 mm thick) with pre-cooled aluminum blocks, all within 120 ms. By this method, the anatomical structures of the sample were well preserved. Transmural metabolism and flow distribution were instantaneously fixed and high resolution of the two-dimensional redox state was obtained by application of NADH fluorescence photography. Micro-samplings from the desired portion of the cross-sectional slice were possible at -190 degrees C. NADH fluorescence of the samples did not increase from the surface to 1.2 mm in depth, confirming that there was no ischemic artifact. With the present technique, a heart sample in which transmural metabolism, and the redox state, are fixed and visualized is attainable, thus providing a new tool for the study of myocardial ischemia.

Adenine Nucleotides↗
PubMed1989

Prolonged survival of mice with myeloid leukemia by subcutaneous injection of recombinant human G-CSF.

We studied the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on leukemia development and survival of mice with leukemia by using a radiation-induced myeloid leukemia cell line (C2M) with A-type phosphoglycerate kinase (PGK) as marker isoenzyme. C3H/He mice with B-type PGK were inoculated with 2 x 10(6) C2M cells through the tail vein. From the next day, they received a daily subcutaneous injection of 1 microgram rhG-CSF or control solution. The survival of rhG-CSF-treated recipients of C2M was significantly longer than control-solution-treated recipients. In rhG-CSF-treated recipients, not only spleen weight but also the number of blasts in hemopoietic organs was less than those in control recipients. While there was a remarkable decrease in bone marrow content of CFU-GM in control recipients, the content in rhG-CSF-treated recipients was comparable to that in normal mice. A reduced bone marrow and spleen content of leukemic colony-forming cells (L-CFU) was observed in rhG-CSF-treated recipients in comparison with control recipients. The electrophoretic analysis of PGK phenotypes of hemopoietic organs indicated the delayed appearance of A-type PGK from C2M cells in rhG-CSF-treated recipients. From these findings, we concluded that the injection of rhG-CSF delayed the onset of leukemia and improved the survival and several hematological parameters by suppressing C2M cells in recipient mice.

Animals↗
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↗
PubMed1989

Effects of recombinant human G-CSF and GM-CSF on primary human leukemic cells.

The effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on primary human leukemic cells were studied. Phagocyte-depleted mononuclear cells containing more than 88% blasts were obtained from peripheral blood of 11 AML and 2 ALL patients and from bone marrow aspirates from 2 ALL patients. The leukemic cells were incubated with these CSF in suspension cultures or in methylcellulose cultures. In suspension cultures, the spontaneous proliferation was observed in 1 M4 patient. RhG-CSF stimulated the leukemic cell proliferation in 5 AML, cases and rhGM-CSF that in 4 AML cases. In methylcellulose cultures, spontaneous colony formation occurred in 3 M4 patients. RhG-CSF and rhGM-CSF stimulated the leukemic colony formation in 8 AML cases. The CSFs had an additive effect in both cultures. Neither CSF induced O2- production or phagocytic activity. From these results, we concluded that both CSFs stimulated the proliferation of leukemic cells without inducing differentiation.

Acute Disease↗
PubMed1988

A new cell line of murine myeloid leukemia with A-type phosphoglycerate kinase as marker isoenzyme.

A new murine myeloid leukemia cell line (C2M) with A-type phosphoglycerate kinase (PGK) as marker isoenzyme was established from myeloid leukemia which arose in a female C3H/He strain mouse of the genotype Pgk-1a/Pgk-1b 1 yr after a whole body X-irradiation of 3 Gy. Cytochemical stainings indicated that C2M cells had myelomonocytic characteristics. Chromosomal analysis showed the partial deletion of No. 2 chromosome. Intravenous injection of C2M cells resulted in the development of myeloid leukemia in syngeneic mice owing to the growth of C2M cells. When C2M cells were transplanted to C3H/He mice with B-type PGK, PGK of spleen expressed two bands on electrophoresis; A-type PGK from transplanted C2M cells and B-type PGK from recipient mice, and the density of A-type PGK became prominent as the disease progressed. When granulocyte/macrophage progenitor cells of bone marrow cells from leukemic mice were cultured, two types of colonies were observed. By determining PGK types of the colonies, leukemic colonies could be differentiated from normal granulocyte/macrophage colonies. Since C2M cell line has an advantage of processing A-type PGK which can be readily distinguished by the electrophoresis from normal cells, it will serve as a useful tool to study the interaction between leukemic cells and normal hematopoietic cells.

Animals↗
PubMed1988

Heritable rare fragile sites in patients with leukemia and other hematologic disorders.

Fragile site studies were performed on a total of 126 patients with leukemia and other hematologic disorders including myelodysplastic syndrome (MDS) and polycythemia vera (PV). Compared with an incidence (6.0%) of heritable rare fragile sites in the healthy population, the frequency was not higher in the patient group (3.2%), as a whole. However, two cases of fra(17)(p12) in MDS appeared fourfold larger than expected for this group of patients. In one case, a homozygous carrier of fra(17)(p12) in PV was also very rarely expected from its population incidence. These findings suggested a possible role of rare fragile sites, at least in the etiology of these preleukemic or myeloproliferative disorders.

Adult↗
PubMed1988

Assay of rat plasma pyruvate kinase activity with luciferin-luciferase.

An assay method for pyruvate kinase in rat plasma is described. Plasma samples were incubated with ADP and phosphoenolpyruvate in Tris buffer solution. The ATP produced by pyruvate kinase was measured by photocounting after the addition of a commercially available luciferin-luciferase preparation. Interference by ATP or adenylate kinase originally present in the sample was removed by a high degree of dilution. The assay is sensitive, reproducible, and rapid, especially when used for large numbers of samples. By this method, pyruvate kinase activity in normal rats was determined to be 0.51 +/- 0.05 (n = 6) U/ml plasma. In rats fed a vitamin E-deficient basal diet for 7, 10, or 14 weeks, pyruvate kinase activities were 0.70 +/- 0.11, 1.64 +/- 0.51, and 4.28 +/- 0.85 (n = 6) U/ml plasma, respectively. This method appears to be useful for the determination of pyruvate kinase activity in nutritional or pharmacological studies.

Animals↗
PubMed1987

Clonal origin of radiation-induced myeloid leukemia in mice with cellular mosaicism.

The clonal origin of radiation-induced myeloid leukemia was studied in mice with cellular mosaicism for phosphoglycerate kinase (PGK). A single whole-body X-irradiation of 3 Gy resulted in development of 11 myeloid leukemias in 198 mosaic mice (5.6%). PGK of spleen cells from 9 myeloid leukemias assayed gave only a single band on electrophoresis. PGK of red blood cells (RBC) from one myeloid leukemia gave a single band identical to that of PGK of leukemic cells, while PGK of RBC from 4 other myeloid leukemias gave two bands. From these results, we concluded that radiation-induced myeloid leukemias in mice were of single cell origin, and that the origin of some myeloid leukemias might be a progenitor cell capable of differentiating into both erythroid and myeloid series.

Animals↗
PubMed1985

Conditions affecting clonal growth of lymphoma cells in a semisolid matrix.

This study demonstrated the importance of the methods used in determining the lymphoma cell colony stimulating activity of factors derived from lymphoma cells. The in vitro colony formation in a semisolid matrix of the AKR mouse lymphoma cell line, SL 12, and three cloned derivatives, SL 12.1, SL 12.3, and SL 12.4, was studied. We show that the use of soft agar or methylcellulose as a semisolid matrix results in colony formation by the lymphoma cells only in the presence of serum. The addition of conditioned medium (CM) from lymphoma cells growing in serum-free medium does not stimulate colony growth. However, when purified agarose is used, colonies grow in a dose-dependent manner in the absence of serum and in the presence of CM. These results indicate that the type of semisolid matrix used can influence results in studies of this nature. Purified agarose provides the best environment when colony formation by lymphoma cells is used to measure the presence of growth factors in test-conditioned media.

Agar↗
PubMed1984

Comparison of the biological activity of two sources of colony-stimulating factor (CSF).

Two possible standard sources of colony-stimulating factor (CSF) have been available commercially; one is GCT-conditioned medium (GCT-CM) and the other is T3M-5A-CSF. To clarify their biological activity and determine whether they can provide a stimulus identical with leukocyte feeders, the number and the morphology of colonies stimulated by them were compared. A dose-response study and a mixing experiment between them suggested the presence of inhibitor(s) in GCT-CM but not in T3M-5A-CSF. Both types of CSF stimulated mainly granulocytic colonies at day 7. T3M-5A-CSF caused a dose-dependent increase in macrophagic colonies at day 14. The number of colonies stimulated by GCT-CM and T3M-5A-CSF did not differ from that of leukocyte feeders, but the morphology of colonies differed significantly. These results indicate that there is a qualitative difference in the effect of GCT-CM and T3M-5A-CSF on human bone marrow cells, and that they do not supply a stimulus identical with leukocyte feeders. Though GCT-CM and T3M-5A-CSF are expected to serve as useful tools in the standard CFU-C assay, we should observe their characteristics and accumulate more information about their behavior on normal as well as on patient bone marrow cells.

Bone Marrow↗