Peripheral leukocytes from 10 patients with CML were cultivated in Petri dishes using RPMI 1640 supplemented with 20% fetal calf serum and their morphological observations were carried out for up to 1.5-2.5 months. Although the cell population per dish generally showed a gradual decrease, cells from all the ten patients began to show a marked maturation to mature basophils at about three weeks after culture initiation and in most cultures these basophils constituted more than 80% of floating cells when studied after five weeks of cultivation. It was unsuccessful to make analyzable metaphases from these basophils but presumably they are derived by maturation of CML cells in vitro.
A patient with CML who presented with a profound increase of basophils and eosinophils during her clinical course is illustrated to show that under certain circumstances CML cells mature preferentially along the basophilic series in vivo.

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Low platelet fibrinogen levels have recently been regarded as one of the important laboratory findings in diseases associated with a hemorrhagic tendency such as thrombasthenia. Because, however, the determination of platelet fibrinogen levels requires a very complex procedure, the determination of the levels has been made in only few cases in Japan. A simplified method for the determination of platelet fibrinogen levels, using the FDP kit, has been successfully developed in our laboratory; and its procedure, factors influential over the results of the determination by this method, and results of its clinical application to a few diseases are presented hereunder.
Normal platelet fibrinogen levels as determined by this method were in the range of 80-320 μg/ml (platelet count: 100×10⁴ cells per cubic mm), while platelet fibrinogen levels in thrombasthenia were as low as 10-20 μg/ml, and those in chronic granulocytic leukemia and primary thrombocythemia were within normal range, ie, 80-160 μg/ml. A decrease in platelet count to one half was paralleled by a drop in platelet fibrinogen level to one half. Washing the platelets seven times eliminated the influence of plasma fibrinogen. This method, the authors believe, is a useful, simplified method for the determination of platelet fibrinogen levels.

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Consecutive bone marrow biopsies were performed 67 times on 54 patients with various hematological and nonhematological disorders. Specimens were obtained in all cases except for one which later found to have aneurysmal bone cyst. Procedures were much easier than with the Vim-Silverman needle. There was no complication.
Specimens were 2 mm in diameter and 10 mm in average length. Only 9 out of 66 specimens (14%) were inadequately small (less than 4 mm in length), and 2 out of 66 specimens (3%) had severe crushing. Otherwise, good specimens were obtained even in patients with hypocellular bone marrow. Indication for bone marrow biopsy was discussed, and four cases in which bone marrow biopsy was clinically useful were reported.
We conclude that this needle is safe, easy to handle, and useful in obtaining good bone marrow specimens.

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In order to assess the diagnostic and prognostic value in preleukemia, marrow cluster and colony forming capacity using soft agar culture technique and maturation capacity in myeloid cells using liquid culture technique were studied in patients with myelodysplasia, potentially preleukemic hematologic diseases and acute myelogenous leukemia (AML).
The cultured marrows in soft agar were classified into five types on the basis of incidences of cluster and colony forming cells, and averaged colony size. (1) Type A; incidences and size in normal controls, (2) Type B; increased incidences of both clusters and colonies with normal or larger colony size, (3) Type C; increased incidence of clusters, and normal or increased incidence of colonies with smaller colony size, (4) Type D; normal or higher incidence of clusters and decreased incidence of colonies, and (5) Type E; decreased incidences of both clusters and colonies.
Although all of the abnormal types (Type B through E) were noted in the agar culture from 32 cases of AML, Type C and D could be regarded as the characteristic types in this disease. Marrow cells from all of 7 patients with myelodysplasia showed abnormal culture patterns; 2 in Type C, 2 in Type D and 3 in Type E. Of these 4 subsequently terminated in AML, and the marrow culture from these patients exhibited 2 Type D, 1 Type C and 1 Type E. The case with Type C remained in the same culture pattern even after subsequent conversion to AML, while the patient with Type E changed the pattern to Type C after transformation into AML. In another patient with myelodysplasia and one with paroxysmal nocturnal hemoglobinuria whose marrow culture showed Type C, subsequently progressing hematological abnormalities to AML developed. The Type E pattern was observed in the marrow culture from most patients with aplastic anemia and one of two patients with sideroblastic anemia besides some patients with AML and myelodysplasia.
In a parallel study of the bone marrow by the liquid culture technique, all materials from myelodysplasia with Type C and D in the agar culture showed a characteristic maturation defect in myeloid cells, while marrow cells from myelodysplasia with Type E showed a normal maturation pattern.
Our present data suggest that the detection of either Type C or D by the marrow agar culture and maturation defect in myeloid cells by the liquid culture in patients suspected of preleukemia may indicate an early sign of leukemic transformation, whereas the observation of Type E in myelodysplastic patients may be of little prognostic value although it suggests certain qualitative alteration of myeloid-committed stem cells.

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A case with monocytic leukemia (Naegeli type) is described. This 44-year-old male patient was admitted to our hospital with chief complaints of generalized skin lesions and oppressive feeling in the upper abdomen. Physical findings included generalized lymphadenopathy involving tonsils, hepatomegaly and splenomegaly, associated with miliary, slightly raised light- brown skin lesions throughout the body.
Hematological findings revealed slight degree of anemia (369×10⁴/mm³), severe thrombocytopenia of 5.34×10⁴/mm³ and leukocytosis of 5.8×10⁴/mm³ with 85% atypical cells. The per cent of the same atypical cells in the bone marrow was 85.2%. Light microscopy revealed cells with monocytic morphology, including 10% of cells with weakly positive peroxidase. The atypical cells are large, measuring 20 to 25μ in diameter, with pseudopodia, nuclei with indentations or folds and bluish cytoplasm. Electron microscopic studies demonstrated a few clearly defined, peroxidase-positive granules in the cytoplasm of the majority of monocytoid cells, consistent with features of monocytic leukemia, Naegeli type. The biopsy of the skin lesion demonstrated infiltration of leukemic cells.
This patient was treated with five courses of multiple combination chemotherapy (three courses; DM, CA, Neocarzinostatin, Pred. two courses; DM, CA, VCR, EX, Pred.) and entered complete remission, concomitant with disappearance of skin lesions all over the body.
It is suggested that electron microscopic studies coupled with cytochemical staining such as peroxidase reaction may be valuable for differential diagnosis between acute monocytic leukemia, Naegeli type and leukemic reticuloendotheliosis, acute type.

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An Autopsy case of extranodal malignant lymphoma was reported. A 49-year-old male was admitted for evaluation of multiple subcutaneous tumors. Neither superficial lymphadenopathy, hepatosplenomegaly, jaundice, nor anemia was present. Laboratory data showed elevated LDH and positive CRP. Chest X-ray, EKG, scintigram of liver and spleen, lymphangiogram were all within normal limits.
Pathological diagnosis of skin tumor biopsy was reticulum cell sarcoma.
Clinical diagnosis was malignant lymphoma, histiocytic type, Stage IV_EA.
The patient was treated with vincristine, cyclophosphamide and prednisolone. Subcutaneous tumors were completely regressed after three courses of chemotherapy, however, III, VI. IV. X. XI. XII cranial neuropathy appeared. The patient expired in a deep coma and bulbar palsy after five months from onset.
Autopsy revealed reticulum cell sarcoma involving subcutaneous, mesenterical and pericadial fat and leptomeninges. Lymphnodes, bone marrow, liver and spleen were intact.
Frequency, primary sites, oncogenes, form of spreading, therapy, and prognosis of extranodal malignant lymphama were discussed.

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A 47-year-old female case with prolymphocytic leukemia of T-cell origin, accompanied by probably drug-induced hepatic injury, was reported. The leukemic cells in the peripheral blood and the bone marrow had characteristic notched, convoluted or lobulated nuclei, which resembled those of Sezary cells. The identification of the leukemic T-cells was performed with four different surface markers for either T- or B-cells. The leukemic cells in the peripheral blood were never responded to corticosteroid and vincristine therapy. Neither anemia nor thrombocytopenia were present. In contrast to hepatosplenomegaly and skin involvement by neoplasm, peripheral lymphnode enlargement was absent. This case was considered to be classifiable into the comprehensive concepts of “adult T-cell leukemia” (Takatsuki et al., 1975) and “cutaneous T-cell lymphoma” (Lutzner et al., 1975).
The post-mortem examination revealed the histologic features resembling malignant histiocytosis and the infiltration of Hodgkin cell like and Reed-Sternberg cell like giant cells to retroperitoneal lymphnodes, liver, spleen and bone marrow. These findings may represent a specific pattern in prolymphocytic leukemia of T-cell origin, rather than the result from therapy.

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A 72-year-old woman was admitted with complaints of nasal bleeding, visual disturbances and general malaise. Physical examination demonstrated lymphadenopathy and hepatosplenomegaly. Moderate anemia and moderate thrombocytopenia were present. Small lymphoid cells were increased in the peripheral blood and in the bone marrow. Paper electrophoresis of the serum revealed a monoclonal peak in the φ region. A diagnosis of macroglobulinemia was made by an increase of IgM in the serum and M-bow against anti-μ on immunoelectrophoresis. Plasmapheresis equivalent 1200 ml of blood was performed during 21 days of hospitalization with little clinical benefits. Incubation of serum at 56°C for 30 min. resulted in formation of opaque gelatinous material, which did not dissolve at 100°C. Cryoglobulin was also demonstrated when the serum was incubated at 4°C for 24 hrs. When pyroglobulin and cryoglobulin were removed from the serum, the M-component disappeared from the resulting supernatant, while it was found in cryoglobulin. Ultracentrifugation analysis demonstrated that the serum M-component and cryoglobulin were 19S macroglobulin. In addition, separate and distinct peak were found to exist both in the serum and in the cryoglobulin which were suspected to be an polymer of 19S globulin. It was suspected that the polymer was related with formation of the pyroglobulin and the cryoglobulin.

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The 8-year-old boy had been repeatedly hospitalized since November 1968 because of intermittent high fever, associated with exanthem. Every admission failed to establish a definite diagnosis and to improve clinical symptoms inspite of steroid administration. To evaluate further his disease, he was admitted to Nagoya University Hospital in October 1973. Hepatomegaly, splenomegaly, moderate hepatic dysfunction and leukocytosis (11,300/cmm) including 48% atypical lymphoid cells without anemia or thrombocytopenia were the remarkable findings in physical and laboratory examinations. On May 20 1974, splenectomy and liver biopsy were performed because of rapidly increasing splenomegaly and development of pancytopenia. The portal pressure was normal. The histology of the spleen showed atrophy of the white pulp and infiltration of lymphoid cells as well as of histiocytes, plasmacytoid cells and “reticulum cells” mainly in the red pulp, with erythrophagocytosis in some of the histiocytes. Liver biopsy also revealed diffuse scattering of mainly lymphoid cells in the sinusoids and portal areas. Soon after the surgery, high fever developed and persisted. Liver was found to enlarge rapidly, accompanied by generalized exanthem. At that time, atypical lymphoid cells were increased gradually in the peripheral blood as well as in the bone marrow. Cytological studies including electron-microscopy and phase contrast microscopy were done. The skin biopsy showed infiltration of the same atypical cells as those in the peripheral blood. In July 1974, combination chemotherapy (VEMP and VENP) was started. He was discharged at the time when a transient remission was achieved clinically, not hematologically. On June 4 1975, he died unexpectedly after a short period of high fever.
The case was presented as a form of “leukemic reticuloendotheliosis” of relatively chronic course developed in a child, which exhibited rather unusual features in many respects.

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