Primary malignant lymphoma of the central nervous system presenting with ascites and pleural effusion.

A 70-year-old woman was admitted to our hospital owing to ascites and pleural effusion. Though malignant cells (B-cell type lymphoma) were detected in both the ascites and pleural effusion, neither lymph node swelling nor a tumor was detected upon chest, abdominal and pelvic computed tomography (CT). After weekly THP-COP therapy for 8 weeks, the ascites and pleural effusion completely disappeared. Two years after the first admission, she was re-admitted because of a disturbance of consciousness, and a brain tumor was detected on CT scan. The immunohistological and genetic data for the brain tumor were identical to those of the malignant cells in the pleural effusion and ascites detected 2 years previously. Whereas the symptoms at onset of a primary lymphoma of the central nervous system (CNS) are usually neurological ones, in this rare case of primary CNS lymphoma, the symptoms at onset were the ascites and pleural effusion without neurological symptoms.

Nuclear accumulation of p53 protein and apoptosis induced by various anticancer agents, u.v.-irradiation and heat shock in primary normal human skin fibroblasts.

Relationship between nuclear accumulation of p53 protein and apoptosis induced by both DNA damaging agents (ultraviolet-irradiation and anticancer agents) and heat shock in normal human skin fibroblasts was investigated. Nuclear accumulation of p53 protein and apoptosis were induced dose-dependently by u.v. -irradiation and anticancer agents. Regarding heat shock treatment, intensive nuclear accumulation of p53 protein was induced by treatment at 43 degrees C for 45 min, but, apoptotic cells were never detected. These results indicated that accumulation of p53 protein and apoptosis induced by DNA damaging agents were considered to have a close relationship and the functions of induced p53 protein are different depending on the stimulating factor.

Scleroderma associated with anemia and thrombocytopenia that responded well to cyclosporin.

A 55-year-old woman came to our hospital because of cutaneous sclerosis of the limbs in September 1996, and was diagnosed with scleroderma based on a skin biopsy. In August 1997, the cutaneous sclerosis became progressive (hemoglobin level, 4.3 g/dl; platelet count, 7 x 10(9)/l). The laboratory results were positive for the direct Coombs test, bone marrow aspiration showed a dry tap, and the bone marrow biopsy showed marked fatty marrow. Indium-111 bone marrow scintigraphy showed a markedly decreasing uptake. These findings indicated bone marrow hypoplasia associated with hemolytic anemia. After prednisolone therapy (60 mg) was initiated, the direct Coombs test became negative but the blood cell count did not increase. Then, 300 mg of cyclosporin was initiated and anemia and thrombocytopenia improved. The cyclosporin dosage was gradually decreased and the patient's hematological condition was good, although the cutaneous sclerosis changed only a little. This is a rare and interesting case of a patient with scleroderma associated with bone marrow insufficiency and hemolysis who responded well to cyclosporin.

Effect of granulocyte/colony-stimulating factor on the onset of the adult respiratory distress syndrome.

To evaluate the effect of granulocyte/colony-stimulating factor (G-CSF) on the onset of the adult respiratory distress syndrome (ARDS), we investigated whether the incidence of ARDS due to pulmonary infection differed between the G-CSF group which received chemotherapy with G-CSF and historical controls without G-CSF. We evaluated 132 patients with hematological malignancy in complete remission without any main organ dysfunction who had been treated between April 1983 and December 1997. We compared the incidence of ARDS due to pulmonary infection between those who received G-CSF and those who did not. There was no remarkable difference in the number of patients, gender, age, or distribution of primary diseases between the two groups. The intensity of chemotherapy was not considered to significantly differ between the two groups, though the chemotherapy regimens administered differed slightly. In the G-CSF group, the duration of neutropenia was significantly shorter and the frequency of documented infection was significantly decreased. We could not find any relationship between ARDS due to pulmonary infection and any anticancer agent or antibiotics. There was no relationship between the kind of G-CSF and the incidence of ARDS due to pulmonary infection (per chemotherapy session; p > 0.10, per case; p > 0.30, chi2 test). The incidence of ARDS due to pulmonary infection per chemotherapy session was 4.21%, and showed a higher tendency in the G-CSF group (p < 0.100, chi2 test). The incidence of ARDS due to pulmonary infection per case was 25.4% and was significantly higher in the G-CSF group (p < 0.025, chi2 test). The incidence of ARDS due to pulmonary infection was higher in the G-CSF group than in the controls, suggesting that G-CSF promotes the development of ARDS due to pulmonary infection.

Effects of daunorubicin on cell growth, cell cycle and induction of apoptosis in HL-60 cells.

Administration of daunorubicin (DNR) at over 3.5 x 10(-8)M has been reported to block cells at G2 phase, but the precise mechanism of cell death induced by DNR is not well known. In this study effects of DNR at various concentrations on cell growth, cell cycle and induction of apoptosis in human leukemia cell line HL-60 cells were investigated. Administration of DNR at a high concentration (3.0 x 10(-8)M) inhibited cell growth to 3% as compared with untreated cells, blocked the cell cycle at G2 phase and induced cell-cycle non-specific apoptosis. Administration of DNR at a lower concentration (1.0 x 10(-8)M) inhibited cell growth to 77%, induced cell-cycle nonspecific apoptosis but did not produce G2 arrest. A longer duration of exposure to DNR was required to induce apoptosis by the lower concentration of DNR than by the higher concentration. These findings indicate that the effect of DNR on cell growth is caused by both G2 arrest and induction of apoptosis and that DNR induced apoptosis without G2 arrest. Moreover, continuous administration of a low concentration of DNR which has been considered to have no anticancer effect might be useful for treatment of leukemia and related diseases.

[DNA and RNA contents of megakaryocytes using acridine orange staining].

We developed a new microfluorometric method to measure the DNA and RNA contents of individual megakaryocytes using acridine orange (AO) staining in human bone marrow smears. Some bone marrow smears were fixed with ethanol and treated with pretreatment solution and then stained with an AO staining solution. The DNA content was assayed by measuring the green fluorescence and the RNA content was assayed by measuring the red fluorescence under B excitation with microfluorometer using peripheral lymphocytes as the control. The ploidy peak was shown to be 16N in all of 5 normal controls, but it was 8N in the patient with refractory anemia with excess of blasts in transformation (RAEB-T). There was not difference in the RNA content/DNA content ratios (RI) between each ploidy in the normal controls. The RI of the patient with RAEB-T was lower than that of the normal controls. The measurement of the DNA-RNA contents may be useful as a new megakaryocytic parameter.

Comparison of the DNA content of megakaryocytes identified immunologically with that identified morphologically.

We devised a new microfluorometric method for determining the ploidy of megakaryocytes identified immunologically in bone marrow smears. The smears were immunostained by incubation with mouse monoclonal anti-glycoproteins (GP) IIb antibodies, followed by fluorescein isothiocyanate-conjugated goat antimouse IgG antibodies. They were then stained with 4',6-diamidino-2-phenylindole (DAPI). Megakaryocytes were identified by their GPIIb immunofluorescence using a microfluorometer and, after the filters were changed, their DNA content was assayed by measuring the intensity of DAPI fluorescence. This intensity was shown to be proportional to the DNA content when the aperture of the objective lens was reduced. We compared these results with those obtained when megakaryocytes were identified morphologically, using DAPI staining after Wright-Giemsa destaining. In all 12 normal controls, the ploidy peaks were shown to be 16N by both methods, and the mean ploidy detected by the immunological method was only reduced 0.961 times relative to the estimate from the morphological method. In contrast, in eight myelodysplastic syndrome (MDS) patients, the ploidy peaks were either 8N or 4N and the mean was reduced by 0.906 times (P = 0.018). Thus we could immunologically identify small megakaryocytes which we could not identify morphologically. Therefore, this method is useful for measuring megakaryocytic ploidy, especially in the pathological megakaryocytes of MDS patients.

[Epstein-Barr virus-infected T-cell malignancy in an adult patient with Behçet's disease-like symptoms].

A 20-year-old woman was hospitalized on November 11, 1994 with Behçet's disease-like symptoms (fever, genital ulcer and aphtha in the oral cavity). Bilateral cervical lymph node swelling was also noted and diagnosed as lymphadenitis on biopsy. Chronic active Epstein-Barr virus infection (CAEBV) was diagnosed based on the high titer of antibodies to the EBV capsid antigen, early antigen, and nuclear antigen. She was treated with prednisolone and acyclovir and all symptoms improved. However, ten months after onset of symptoms, T-cell malignancy was diagnosed on bone marrow aspiration, which revealed 34.9% blast cells that had rearrangement of TCR-beta. She died on May 8, 1995, despite anticancer therapy. In analyzing the blast cells, the monoclonal junctional DNA structure of the EBV terminal repeat was analyzed by Southern blotting and provided definitive evidence for the monoclonality of EBV-infected T cells. These findings strongly suggest that EBV plays a pathogenic role in T-cell malignancy. EBV-infected T-cell malignancy, such as this case, is very rare in Japan, especially in adult.

[Infections mononucleosis with pleural effusion].

A 24-year-old man was admitted to our hospital for further examination of pleural effusion. On physical examination, he had a temperature of 39 degrees C, the pharynx was painful and liver and spleen were enlarged. The leukocyte count was 5,700/microliters (atypical lymphocyte 6%). The serum LDH, GOT, GPT, ALP and gamma-GTP levels were elevated, and antibodies to Epstein-Barr viral capsid, early, and nuclear antigens were diagnostic of a primary Epstein-Barr virus infection. The CD4/CD8 ratio of peripheral blood lymphocyte was decreased to 0.2. The pleural effusion was exudate, and infiltration of mononuclear cells was noted. The CD4/CD8 ratio of lymphocytes in the effusion also was decreased to 1.1. The result of pleural biopsy showed a perivascular infiltration of mononuclear cells and immunological stain showed that the infiltrated cells were dominantly T-lymphocytes (about 90%). These findings suggested that the pathogenesis of pleural effusion in infectious mononucleosis was a pleulitis due to the infiltration of T-lymphocytes. Pleural effusion is known as a rare complication of infectious mononucleosis.

Primary acute myeloid leukemia cells negative for c-kit receptor protein expression do not proliferate in vitro in response to colony-stimulating factors.

This study examined the effects of recombinant human stem cell factor (SCF) alone or in combination with colony-stimulated factors (CSFs) on colony formation of leukemic progenitors obtained from 29 acute myeloid leukemia (AML) patients. C-kit receptor (c-kitR) protein expression was examined using an immunofluorescence method and was detected on more than 10% of leukemic cells in 20 of 29 cases (c-kitR+). SCF alone could stimulate formation of colonies in 14 of 20 c-kitR+ cases. Granulocyte(G)-CSF, granulocyte-macrophage (GM)-CSF, and interleukin (IL)-3 alone stimulated colony growth in 18, 17 and 16 of c-kitR+ cases, respectively. In contrast, colony and cluster formations from eight of nine c-kitR- cases was not stimulated at all by SCF alone nor SCF in combination with CSF nor with any CSF alone. However, the in vitro culture behavior of fluorescence-activated cell sorter-sorted c-kitR- cells from c-kitR+ cases did not significantly differ from c-kitR+ cells. These results suggest that responses of leukemic progenitors to CSFs were variable but may be predicted by their c-kitR expressions, and that c-kitR(-) cases may have a different disease entity from c-kitR(+) cases.