Isozyme of acid phosphatase in human leukocyte with a variety of hematological diseases were examined. Six isozymes, namely 1, 2, 3, 3b, 4 and 5 were separated by acrylamide disc gel electrophoresis. The study of these isozymes in preparations of various cell types indicates that some of the isozymes were cell type specific, i. e., isozyme 1, 2, 3 and 4 in neutrophils, 3 in lymphocytes and tumor cells of lymphoproliferative disorders, 4 in monocytes and 3b in blast cells irrespective of cell series.
Although leukocyte preparation of hairy cell leukemia showed isozyme 5 which is tartrate resistant, considerable quantity of this isozyme was also demonstrated in some of other hematological malignancy cells, especially of CLL and RCS cells.

View full abstract

Since 1973, a total of 27 children (18 with acute lymphocytic leukemia (ALL) and 9 with acute myelocytic leukemia (AML)) have been treated with “Total Therapy” protocol Study 7 and/or Study 8, which was presented by the Hematology Service in St. Jude children's Research Hospital, Memphis, Tenn. This treatment entails four phases, 1. remission induction by prednisone and vincristine, 2. preventive CNS therapy by ⁶⁰Co 2400 rads cranial irradiation with intrathecal methotrexate or craniospinal irradiation, 3. maintenance chemotherapy by 6MP, MTX and cyclophosphamide and 4. intensive chemotherapy by prednisone and VCR.
Complete remission was obtained in 94% of ALL and in 56% of AML. After successful remission induction, 17 patients received preventive CNS therapy and it was only 2 patients that developed CNS leukemia.
13 patients (11 in ALL and 2 in AML) have now been in initial continuous complete remission. The longest duration of complete remission is 42 months.
Toxicity of the preventive CNS therapy was observed in 86% of the patients, who complainted nausea, vomiting, headache, fever and CSF pleocytosis. 3 patients developed acute but tentative chemical miningitis due to MTX, a few hours after intrathecal injection.

View full abstract

Acid phosphatase cytochemistry was performed on the lymphocyte subpopulation obtained from human peripheral blood.
On cytochemical study, distinct difference was observed between E-Rosette forming cells (RFC) and non-E-rosette forming cells (non-RFC).
Acid phosphatase activity was stronger in RFC than in non-RFC. Few large coarse granules of reaction materials were observed about 41% of RFC in the vicinity of Golgi area, only 5% of non-RFC.
On the other hand, no or some small fine granules were observed in non-RFC and the reaction materials were dispersed in the cytoplasma.
The thymocytes obtained from open cardiac surgery, as T-cell model, have shown few localized large coarse granules in the cytoplasms.
On the cytochemical study, L(+)-Tartaric acid resistant acid phosphatase activity was shown on the lymphocytes stimulated in vitro with phytohemagglutimin, but never seen in peripheral lymphocytes by cytochemical and isoenzymatic study.
The fact may suggest that acid phosphatase activity is mainly concerned with cell cycle or cell function, not its origin.

View full abstract

Purified platelet aggregating factor (PAF) was obtained from bovine BaSO₄-adsorbed plasma through DEAE cellulose column after bentonite adsorption and polyethylene glycol fractionation. This purified PAF was stable to heat (54°C, 150 min.), and plasmin digestion (1 cu/ml, 120 min.), against the obserbations of Vermylen, J. et al. & Fukui, H. Using this purified PAF, PAF-induced platelet aggregation was studied on human platelets in comparison with Ristocetin-induced platelet agreegation (RIPA).
Five patients with hemophilia A showed normal platelet aggregation with both Ristocetin and PAF. Studied were PRP of three patients with von Willebrand's disease whose Ristocetin cofactor (VIII_VWF) was below 12.5% of normal. In two of them, whose AHF like antigen (VIII_AGN) level was 64%, 49% of normal, PRP showed no aggregation with Ristocetin, but normal aggregation with PAF. However, the other case with von Willebrand's disease, whose VIII_AGN level was not detected, showed hyperaggregation with PAF. It may be suggested that human VIII_AGN have some inhibitory effects on PAF-induced aggregation, through saturation of receptors on platelets.
In five cases with thrombasthenia, the intensity of platelet aggregation with both Ristocetin and PAF was reduced, and their PRP showed spontaneous disaggregation. Addition of ADP (10 μM) inhibited Ristocetin- and PAF-induced aggregation, and ATP (2.5 mM) overcomed these inhibitory effects of ADP. ATP also prevented disaggregation seen in thrombasthenic PRP as well as EDTA treated normal PRP. Although addition of ADP to formalin fixed normal platelets did not inhibit RIPA, aggregation was reduced when ADP was added before these EDTA treated platelets were fixed. ATP prevented this inhibitory effects of ADP, too. These findings support the reports of Cohen, I. et al. & Grant, R. A. et al, and might suggest that ADP and/or ATP bind to platelet membrane so that the receptors to von Willebrand factor, or PAF may become transformed.

View full abstract

There have been only a few reports on leukemia in married couples. Recently we encountered two couples with acute leukemia: the first couple was a husband with acute promyelocytic leukemia and his wife, still living, with acute monocytic leukemia, and the other a husband with acute monocytic leukemia and his wife with acute lymphocytic leukemia, who died 3.5 years prior to the onset of the disease of her husband.
At present it is very difficult to determine whether or not simultaneous occurrence of leukemia in couples is more than the chance. The accumulation of data of such cases will throw a light on the study of epidemiology of leukemia.

View full abstract

Two cases of erythroid hypoplssia caused by Tegretol treatment were reported. Case 1 was a 37-year-old man who showed serious anemia due to the therapy for traumatic epilepsy with Aleviatine (Diphenylhydantoin) and Minoaleviatine (Trimethadion). After discontinuation of the drugs the symptoms were relieved only by blood transfusion. Then Tegretol was administered 400 mg per day for 7 months to a total amount of 42.4 g. Case 2 was a 44-year-old man with aneurysm of anterior communicating artery. After clipping the aneurysm, Tegretol was administered 600 mg per day for 4 months to a total amount of 96.2 g, for preventing postoperative convulsion and suppressing psycological symptoms. In both cases, only the count of erythrocyte serious decreased but the count of leukocyte and platelet were mild decreased or almost normal in the peripheral blood. In the bone-marrow, only erythroblasts were selectively damaged. Within 4-6 weeks of the discontinuation of Tegretol, anemia was improved.

View full abstract

A 45-year-old man with Coombs-positive hemolytic anemia induced by α-methyldopa therapy for 21 months duration was described.
The clinical picture consisted of slight jaundice and splenomegaly. The laboratory examination disclosed Hb 11.6 g/dl, RBC 3.06×10⁶/mm³, reticulocyte 17.2%, serum bilirubin 6.4 mg/dl, erythroid hyperplasia of bone marrow (M: E 0.4), and positive direct and indirect Coombs tests.
The immunoglobulin class of the Coombs antibody was IgG of both κ and λ type, and Rh antigen specificity was not demonstrated.
The withdrawal of α-methyldopa brought a gradual improvement of anemia and jaundice, and the Coombs test became negative 368 days later.

View full abstract

A 68-year-old male was admitted to Okayama University Hospital because of leukocytosis and hepatosplenomegaly. During the four months prior to admission, the patient experienced malaise and easy fatigability. On examination the liver was palpable 3 cm and the spleen 5 cm below the costal margin. The peripheral leukocyte count was 57,600/mm³, 92% of which were large lymphocyte-like cells. Most of these abnormal cells exhibited many hairy cytoplasmic projections, when examined under the phase contrast microscope. These cells were negative for acid phosphatase as well as for tartrate-resistant acid phosphatase. The majority of leukemic cells had IgG immunoglobulin on the cell surface. Electron microscopy revealed the presence of a ribosome-lamellar complex in the cytoplasm. These hematological and physical findings have remained essentially unchanged for 14 months from the onset of symptoms. This case was, therefore, considered to represent a variant type of hairy cell leukemia in that hairy cells lacked acid phosphatase in the presence of all other clinical and cytological characteristics of hairy cell leukemia.

View full abstract

Peculiar inclusion bodies were demonstrated in the myeloblasts of both peripheral blood and bone marrow from a 17 year-old female, suffering from acute myeloblastic leukemia. This patient treated by Daunorubicine, Cytocine-arabinoside, 6-MP and corticosteroids, gained 11 months complete remission and survived 24 months after onset of the illness.
The light microscopy of the myeloblasts with May-Grunwald Giemsa stain showed small red granules, Auer rods, and several vacuoles in the cytoplasm.
By electron microscopy, 39% of the myeloblasts had one or several round vacuolar inclusion bodies, from 0.1μ to 4μ in diameter in the cytoplasm. They were located near the nucleus and had a limiting membrane, and contained fine granules, filaments, crystalloids, or a membranous substance besides a small amount of floccular material. In the cytoplasm of some myeloblasts another type of large electron dense granules were found and they were suggested to be giant azurophilic granules associated with acute granulocytic leukemia.
These vacuolar inclusions were suspected to be possibly secondary lysosomal in nature.

View full abstract

Pernicious anemia (PA) is rare in individuals younger than 20 years of age. The adult form of PA is common in Europeans, particularly in Scandinavian, English, and people of Irish ancestry, but is rather rare in Orientals including Japanese. Nevertheless, PA in childhood is seen in higher rate among Japanese than among Europeans. Up to now, 15 cases under 20 years old have been reported in Japan.
A 14-year-old junior high school girl was admitted to our hospital because of an extreme anemia and a high fever. The anemia was macrocytic and hyperchromic. Bone marrow showed typical megaloblastic erythropoiesis. Serum vitamin B₁₂ level was as low as being not detected with a radioassay. Schilling test was done without and with hog intrinsic factor and showed that the deficiency of vitamin B₁₂ was due to the lack of intrinsic factor. Gastric juice after stimulation with tetra-gastrin contained no free hydrochloric acid nor intrinsic factor activity. Antibodies against intrinsic factor and parietal cells were positive in the serum. No other autoantibodies nor endocrinopathy were detected.
Sixteen cases, including 15 cases ever reported and the present case, were divided in two groups according to the age of onset; the former 8 cases under 4 years and 8 months, and the latter 8 cases from nine to eighteen years. The former included two definitely proved cases of Imerslund syndrome and of “congenital” type of McIntyre et al., respectively. Six cases, including the present case, of the latter group appeared to belong to the “juvenile” type of McIntyre, et al. Five out of these cases showed achlorhydria after stimulation with histamin or tetra-gastrin; three of the five cases were confirmed to be deficient of intrinsic factor with Schilling test. It is an interesting feature that all of these six cases were female and showed no endocrinopathy, which is one of the components of this type.

View full abstract

A 37-year-old male was admitted on May 17, 1973, because of purpura over the extremities. As the platelet count was 2000/mm³ and immature-looking megakaryocytes were increased in the bone marrow, a diagnosis of thrombocytopenic purpura was made. The treatment was initiated with corticosteroid which did not effect much and thereafter thrombocytopenia remained. A month after admission, insomnia developed and thyroid functions revealed T₃ 46.4%, T₄ 12 μg/dl and BMR+13%. Having an episode of subarachinoidal hemorrhage on Aug. 27, 1974, he was readmitted. The further examinations performed after recovery from this episode, revealed that antithrombocyte antibody was negative, antithyroglobulin antibody 100X positive, antimicrosome antibody negative and LATS negative. Tri-iodothyronin supression test showed poor supression and pneumomediastinography showed the enlarged thymus in the anterior mediastinum. Hyperthyroidism fluctuated from relapse to remission without antithyroid treatment and this fluctuation of the disease seemed to correlate with the dose of corticosteroid. The possible relationship between thrombocytopenic purpura and hyperthyroidism was briefly discussed.

View full abstract

The clinicopathological, immunohistochemical and ultrastructural findings of an autopsy case of thrombotic thrombocytopenic purpura (TTP) is reported. A 31-year old woman was hospitalised with jaundice, purpura, fever, albuminuria and hemoglobulinuria. Laboratory examinations revealed hemorrhagic diathesis due to thrombocytopenia and microangiopathic hemolytic anemia characterized by fragmented red blood cells. Soon after hospitalisation, mental disturbance was noted. FDP and fibrinogen in the blood were within normal level, excluding DIC from the consideration. The diagnosis was TTP. High doses of steroid, heparin and dipyridamole were administered. Since the renal dysfunction rapidly progressed, hemodialysis was carried out. However, she was deteriorated progressively and died three weeks after the onset of symptoms. The characteristic histologic features of TTP were obtained by autopsy. The multiple vascular occlusions were found to be present in various organs, especially in the heart and brain. The vascular occlusions were due to the deposits of homogeneous PAS-reaction positive and PTAH staining negative materials in the subendothelial space of mainly arterioles. The aggregated platelets were frequently seen in the vascular lumens. The immunofluorescence and ultrastructural studies demonstrated that the occlusive materials were stained intensely with rabbit anti-human fibrinogen serum and consisted of homogeneous substance intermingled with degraded platelets. Polymerized fibrin was not observed. The granular deposits of IgM and beta-1C globulin were found intramurally in some arterioles.
Ultrastructual study revealed the electron dense granular deposits distinct from the occlusive materials in some arteriolar walls, suggesting the presence of immune-complexes associated with IgM and beta-1C globulin.

View full abstract

A case of 11-year old boy with combined deficiency of factor IX & VIII with prolonged bleeding time, the first case in Japan, was reported. He was admitted because of frequent hemorrhagic episodes such as hemarthrosis, ecchymosis or hematuria since his late infancy. There were on hemorrhagic diathesis found in his family, and parents were not consanguinous marriage.
He had hemarthrosis on both knee joints on admission, when laboratory data revealed markedly prolonged bleeding time (15 minutes) and partial thromboplastin time (247.6 sec.) and characteristic marked decrease of factor IX associated with moderate decrease of factor VIII was found. Ristocetin induced platelet aggregation was poor (5.4%). Factor VIII-related antigen was decreased (25%). Platelet adhesiveness was almost normal. Factor VIII increased more than desired value and its value stayed for over 24 hours following infusion of AHF-MIDORI with improvement of prolonged bleeding time and also elevation of factor VIII and improvement of bleeding time was seen following infusion of PPSB-NICHIYAKU. Factor IX & VIII increased more than desired value after infusion of normal plasma but factor IX decreased rapidly in a short time. In family study mother had the low value of factor IX and showed the same responce as patient for normal plasma infusion. After infusion of normal serum factor IX of patient was not influenced. The results may indicate that this case is suitable to be angiohemophilia AB.

View full abstract