PubMed Health⌕ Search

Biomedical subjects

N Maruo

Publications and source records attributed to N Maruo.

At least 37 records · Page 2Linked to original sources

[Qualitative and quantitative evaluation of CFU-E colonies in myelodysplastic syndromes].

Hemoglobin (Hb) synthesis in colony-forming-unit erythroid (CFU-E) colonies from the bone marrow of 10 normal subjects and 6 patients with myelodysplastic syndromes (MDS) was studied using microcytofluorometry. Hb content was determined utilizing a photochemical reaction in which the intracellular Hb is converted into fluorescent porphyrin. Briefly, the CFU-E in plasma clots was assayed according to Tepperman's method. Bone-marrow mononuclear cells were then dispensed onto petri dishes containing the plasma clots. The cells in plasma clots were cultured for seven days and then air-dried. The samples were then fixed with pure methanol. Thereafter, the dishes were exposed to ultraviolet light (lambda = 405 nm) in the presence of an SH-donor (0.2 M mercaptoethylamine hydrochloride). The intensity of porphyrin fluorescence was measured using a microcytofluorometer. When the photochemical reaction was carried out for 50 min, the intensity of fluorescence was found to be proportional to the mean corpuscular Hb (MCH) level, suggesting that the intracellular Hb level can be determined as a function of the intensity of this fluorescence. The number of CFU-E colonies in MDS patients was smaller than that in normal subjects. In addition, intracolony Hb level was markedly lower in the MDS cases than in normal subjects. Our findings suggest that the maturation process of CFU-E is impaired in MDS patients.

Adult↗

[Acute unclassified leukemia with bone marrow necrosis].

Massive bone marrow necrosis was seen in a 42-year-old male with acute leukemia. In December, 1988, on admission, laboratory data revealed pancytopenia and a high level of serum LDH and ALKP. Bone marrow aspiration resulted in dry-tap and showed bone marrow necrosis in the bone marrow biopsy specimen. A bone marrow scintigraphy with 111In faintly visualized the bone marrow but visualized area was expanded in the extremities compared with normal subjects. The second bone marrow biopsy showed proliferation of blasts. In the middle of March, blasts began to appear in peripheral blood. The blasts were cytochemically negative for POX, Es, PAS, AcP, TdT and had surface markers CD3-, CD19-, CD33-, CD13-, LCA-, HLA-DR-. Even by investigation on rearrangement of the immunoglobulin heavy chain region, an origin of the blasts could not be determined. In April, the number of blasts in peripheral blood increased and hepatosplenomegaly developed rapidly. Therefore, he was put on the chemotherapy with vincristine and prednisolone, but he died of cerebral hemorrhage. The autopsy revealed widespread bone marrow necrosis. It has rarely been reported that massive bone marrow necrosis is found prior to the occurrence of acute unclassified leukemia.

Acute Disease↗

A microcytofluorometric method for quantitative and qualitative evaluation of CFU-E and BFU-E colonies.

A new method has been developed for the precise identification of human bone marrow colony forming unit erythroid (CFU-E) and burst forming unit erythroid (BFU-E) colonies, and for determination of the hemoglobin contents using microcytofluorometry. The method relies on a photochemical reaction in which intracellular hemoglobin is converted into fluorescent porphyrin under violet light (lambda = 405 nm) in the presence of an SH-donor (mercaptoethylamine hydrochloride). The CFU-E and BFU-E colonies showed red fluorescence with two spectrum peaks at 600 and 650 nm when illuminated by violet light. These two peaks are consistent with those of porphyrin fluorescence. The porphyrin fluorescence was not inducible in colony forming unit granulocyte-macrophage (CFU-GM) colonies, while 20% of the CFU-GM colonies were false positive with respect to the conventional benzidine reaction. The photochemically inducible fluorescence began to appear in BFU-E colonies on the 4th day of culture, while the same colonies started to be positive for the benzidine reaction on the 9th day. Therefore, the photochemical reaction was more specific and sensitive than the benzidine reaction for the identification of CFU-E and BFU-E colonies. In addition, this method enabled us to measure the hemoglobin level in the cells forming the colonies because the intensity of the fluorescence was proportional to the amount of hemoglobin when the photochemical reaction was carried out for 50 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

[A case report of an acute myelogenous leukemia (FAB M1) with Klinefelter's syndrome].

Reported is the second documented case in Japan of acute leukemia with Klinefelter's syndrome. The patient, a 52-year-old male, was admitted to hospital on July 9, 1983, after complaining of a fever and general malaise. The hematological examinations revealed a WBC count of 14,300/cmm with 32% being leukemic cells. Bone marrow aspiration disclosed a nucleated cell count of 687,000/cmm with 41.6% being leukemic cells consisting of myeloblasts. The endocrinic laboratory data showed a high level of urine gonadotropin and chromosomal studies revealed a 47,XXY karyotype. A diagnosis of acute myelogenous leukemia (M1) with Klinefelter's syndrome thus was derived from these findings.

Humans↗

[One-year remission of chronic myelogenous leukemia (CML) after discontinuation of interferon-alpha].

A 53-year-old woman was admitted to our hospital on Nov. 16, 1987, because of general fatigue. On admission, she had hepatosplenomegaly and her peripheral blood profile showed a white blood cell count (WBC) of 309 x 10(3)/microliters with immature neutrophils, a hemoglobin level (Hb) of 7.6 g/dl, platelet count (PLT) of 536 x 10(3)/microliters, neutrophilic alkaline phosphatase (NAP) score of 44. Both Vitamin B12 and LDH levels were high. The bone marrow showed marked myeloid hyperplasia. In a cytogenetic study, Ph1 was found in 3 of 8 metaphases and Ph1 with an additional abnormality of 8 trisomy was noted in 5 of 8 metaphases. She was diagnosed as having chronic myelogenous leukemia (CML) and treated by i.m. injection of interferon (IFN)-alpha at a daily dose of 6 x 10(6) U. Administration of IFN-alpha induced fever for a few days. WBC, PLT count and LDH level gradually decreased, and the NAP score and hepatosplenomegaly improved. She achieved remission in February, 1988. Administration of IFN-alpha was stopped in April, 1988, when the bone marrow showed hypocellularity and normal karyotype. She was treated with 20 mg of prednisolone daily from May until August, because of progressive pancytopenia. She had received no treatment until July, 1989. In May, 1989, the bone marrow again showed myeloid hyperplasia and Ph1 was found in all cells analyzed. Therefore, we resumed IFN-alpha treatment. It is interesting that remission of CML continues for more than one year after discontinuation of IFN-alpha in this case.

Female↗

[Measurement of the DNA content in morphologically identified cells on a Wright-Giemsa stained smear].

4',6-Diamidino-2-phenylindole (DAPI) staining was applied to measuring nuclear DNA contents of single cells previously identified on a Wright-Giemsa stained smear. The smear was first photographed and the location of each cell was mapped. The smear was then destained with 50% acid ethanol and absolute methanol, and restained with DAPI (pH 7.4) at 4 degrees C. Blue fluorescence was observed in the nucleus, and the intensity of this fluorescence was proportional to the amount of DNA, with the lowest coefficient of variation of 3.6% in the case of staining the peripheral lymphocytes for 30 min. This method is useful for measuring DNA contents in heterogeneous cell populations.

Azure Stains↗

[Successful bronchial arterial infusion (BAI) of ACNU in the treatment of pulmonary infiltration of acute non-lymphocytic leukemia (ANLL) cells].

A 35-year-old man was admitted to our hospital because of lumbago on March 25, 1988. On admission white blood count was 1,200/microliters with neutrophils of 9% and lymphocytes of 91%, hemoglobin level was 11.2g/dl and platelet count was 55 x 10(3)/microliters. Bone marrow smear showed 77% leukemic cell including non-specific or specific esterase-positive cells. Chest X-rays showed the presence of mediastinal tumor and diffuse reticular shadows. A diagnosis of ANLL was made and a hematological remission was obtained after one course of combination chemotherapy consisting of BH-AC, daunorubicin and prednisolone, but the enlarged mediastinal tumor and pulmonary infiltration worsened rapidly followed by marked dyspnea. This radiographic abnormal shadow was confirmed to be leukemic infiltration from the finding of transbronchial lung biopsy. We hesitated to give systemic chemotherapy because he also had had liver abscess. Accordingly we performed BAI of ACNU at a dosage of 150 mg which led to a dramatic improvement in dyspnea. 60Co therapy was performed on the mediastinal tumor. On May 30, when he had a relapse, he was unsuccessfully treated with systemic chemotherapy. The leukemic cells invaded most of the organs and the patient died on July 19, 1988. It is likely that BAI of ACNU for leukemic pulmonary infiltration was effective.

Adult↗

[Polycythemia vera terminating in myelodysplastic syndrome].

A 77-year-old male, who had been treated with carboquone and busulfan for polycythemia vera (PV), developed myelodysplastic syndrome (MDS) 8 years later. On admission the peripheral blood revealed pancytopenia, but blastoid cells were not noted. The bone marrow showed hypercellularity, and functional and morphological abnormalities in trilineages of hemocytes. Cytogenetic study showed complex abnormalities involving chromosomes 5 and 7. We diagnosed this case as secondary MDS to alkylating agents. He was treated with 1, 25 (OH)2 vitamin D3. However, it was not effective and the percentage of myeloblasts increased to 14.4%. In spite of supportive therapy, he died of sepsis due to urinary tract infection.

Aged↗

[Aplastic anemia associated with minimal serum M-protein treated successfully with repeated bolus methylprednisolone therapy].

A 61 year old woman was admitted to our hospital suffering from severe pancytopenia. Bone marrow aspiration revealed marked hypoplasia with 24.8% plasma cells. There was a small amount of IgA type M protein in serum. Multiple myeloma was ruled out on the basis that 1) the amount of serum IgA-M protein was minimal and no increase was observed, 2) no indication of monoclonality of bone marrow plasma cells was shown by immunofluorescent staining of immunoglobulins, 3) neither bone lesion nor other symptoms related to multiple myeloma were observed. Therefore, a diagnosis of aplastic anemia was made, and bolus methylprednisolone therapy (1,000 mg/day x 3 days) was performed, which brought the patient to complete remission. An oral dose of 15 mg of prednisolone is administered daily now and the remission state is being maintained. Bolus methylprednisolone therapy is performed quite often for the treatment of aplastic anemia. However, there are few reports in which remission was reached with the use of glucocorticoids alone, most having used androgens or other supportive drugs in conjunction. Here, we will discuss this case of aplastic anemia which responded to repeated bolus methylprednisolone therapy, with minimal side effects, and discuss its effectiveness. In addition, aplastic anemia associated with idiopathic monoclonal gammopathy is not reported, so this case is interesting immunologically.

Anemia, Aplastic↗

[Megakaryocyte ploidy in patients with myelodysplastic syndrome--by microcytofluorometry with DAPI staining after removal of Wright-Giemsa staining].

In the myelodysplastic syndrome (MDS) patients the ploidy distribution of megakaryocyte DNA was rarely reported. We applied DAPI (4',6-diamidino-2-phenylindole) staining for measuring nuclear DNA content in megakaryocytes which have been morphologically identified on the Wright-Giemsa stained smear in 8 normal subjects and 12 MDS patients. Briefly, megakaryocytes morphologically examined on a Wright-Giemsa stained smear were photographed, and were located. We then removed the Wright-Giemsa stains by immersing it in 50% ethanol, 37 degrees C for 1 hour and 100% methanol, 37 degrees C for 1 hour. The DAPI staining was performed in DAPI solution (DAPI 0.01 mg/ml, pH 7.4 Tris-EDTA-2 Na buffer solution and 0.01 mol 2-mercaptoethylamine hydrochloride were mixed at the ratio of 0.5: 98.5: 1.0) for more than 30 min. The amount of nuclear DNA in the megakaryocyte previously identified was measured by cytofluorometry. The population of megakaryocyte in the normal subjects was the largest in the 16N, and in the 10 cases of the 12 MDS patients was the largest in the 8N, in the other 2 cases was the largest in the 4N. These results represent the development of the megakaryocyte nucleus in the MDS patients may be disturbed.

Adult↗

[Microcytofluorometrical measurement of DNA content in immunologically identified micromegakaryocytes of human bone marrow].

We have developed a method to microfluorometrically determine the amount of DNA in immunologically identified micromegakaryocytes on bone marrow smears. Bone marrow smears were fixed with acetone-formalin buffer and immunostained with a monoclonal anti-GPIIb/IIIa antibody, followed by FITC conjugated anti-mouse IgG. After the smears were re-fixed with methanol, DAPI (4', 6-diamidino-2-phenylindole) staining was performed. Using an automatic Digital-Microfluorometer (Olympus MMSP-FR-II), megakaryocytes on the smears were identifies by the GPIIb/IIIa immunofluorescence and, after changing the barrier filters, their nuclear DNA content was measured by the intensity of DAPI fluorescence, which is proportional to the amount of DNA. Using this method, we found that the DNA histogram of the megakaryocytes from a patient with myelodysplastic syndrome showed a shift to small ploidy compared with normal controls. This method may be valuable in the measurement of the megakaryocyte DNA content.

Adult↗

[Acute myelogenous leukemia with recurrence of the meningeal, spinal cord and the optic nerve infiltration with remission induction by Co therapy and maintained by intrathecal chemotherapy with Ommaya reservoir].

A 42-year-old woman was diagnosed as having acute myelogenous leukemia (M 2 of FAB classification) in May, 1985. Complete remission was achieved with the BHAC-DP therapy (BHAC, daunomycin and prednisolone). Whole skull irradiation and intra thecal chemotherapy were performed by way of prevention. In 1986, she developed her visual disturbance and paraplegia. Myelography and metrizamide CT indicate intraspinal and meningeal infiltration of leukemic cells in the upper thoracic spine. Leukemic cells were found in the cerebrospinal fluid but not in the bone marrow. Complete remission was obtained by irradiation and intrathecal chemotherapy. In Dec. 1986, Ommaya tubing was repeated. Hematological and neurological examinations showed no evidence of malignancy for 3 years. In Oct. 1988, she admitted to our hospital for the maintenance of the remission state, and died of sepsis. The result of autopsy revealed no leukemic cells in her CNS. It is said the intraspinal infiltration of leukemic cells is rather rare. As it is said the intrathecal injection of anti-leukemic agents is not effective for the intraspinal infiltration, irradiation should be needed. Therefore it is very important to critically differentiated intraspinal from meningeal infiltration. Furthermore, Ommaya tubing is effective in order to maintain remission in a patient with CNS leukemia.

Adult↗