A low-molecular-weight proteinase inhibitor was isolated from healthy human plasma. After diafiltration using an Amicon YM 5 membrane, human plasma were concentrated by an Amicon YM 2 membrane. And then samples were chromatographed on a Sephadex G-25 column. After fractions of inhibitory activity against papain were pooled, they were passed through CM Cellulofine. The eluate was then transferred onto a DEAE Cellulofine column. The column was eluted with 200-ml linear gradient of NaCI. Tubes containing inhibitory activity were concentrated and rechromatographed on a Sephadex G-25 column. The isolated fraction showed a single band on 15 % polyacrylamide gel electrophoresis at pH 9.4, and inhibitory activity against papain and trypsin was detected in the band. Approximately 5μg of the isolated inhibitor caused a 50 % inhibition of 5μg papain, and 5μg resulted in a 50 % inhibition of 5μg of trypsin. The molecular weight of the inhibitor was estimated to be approximately 3, 200 daltons by the gelfiltration on Sephadex G-25.

View full abstract

The effect of atenolol on lipid metabolism in the spontaneous hypertensive rats (SHR) were investigated by the determination of lipid contents in the serum, liver and aorta. Male SHR at 5 weeks of age were used. Rats were given 20 and 100 mg/kg/day of atenolol orally for 10 weeks. Blood pressure and heart rate were inhibited in dependence on a dose of atenolol the whole of the experimental period. Serum triglyceride (TG) were significantly decreased by the administration of atenolol at a dose of 100 mg/kg. Other serum lipids were not changed by the administration of atenolol. The lowering of serum transaminase (GOT) by atenolol at a dose of 100 mg/kg were seen at 2, 4, 8 and 10 weeks of administration. Liver total cholesterol and TG tended to increase at the early weeks of administration at dose of 20mg/kg and later weeks of administration at a dose of 100 mg/kg. Aorta lipid contents were not changed after administration of atenolol. Atenolol caused a mild chages on the lipid metabolism than the other IS-blocking agent (e. g. propranolol) . These results suggest that the continuative treatment of atenolol has a rare posibility inducing the abnormality of lipid contents.

View full abstract

By using lymph node imprints we studied ANAE-positive cell increasing diseases, the both malignant and benign diseases, except malignant lymphoma and non-specific lymphadenitis. In the 6 cases of malignant disease ANAE-droplet positive lymphocytes had higher percentage value than that of chronic nonspecific lymphadenitis group, and 16 cases of benign disease were shown the same result. In general, at the peripheral area of metastatic tumors in the lymph nodes many reports are indicated that lymphocytes are apt to depletion. This report might be an exceptional case group that there is rather prominent esterase-positive T-cell population. In benign disease group they are consisted of 6 cases of tuberculosis, 3 Piringer's lymphadenitis, 2 syphilis and 1 case of eosinophil granuloma, sea blue histiocytosis, infectious mononucleosis, sarcoidosis, respectively. Special diseases in malignant group were included neuroblastoma and Ewing's sarcoma. It is still obscure that ANAE-positive Tcell may have a killer effect against malignant tumor and/or specific inflammatory organism.

View full abstract

By using ANAE staining we reported here its activities of tumor cells in some relative rare diseases of lymph node and spleen. Two cases of hairy cell leukemia were suggested B cell origin with granular positive reaction products.
Three cases of histiocytic sarcoma were shown diffuse reaction products in the whole cytoplasma seen as phagocytic staining pattern.
Five cases of prolymphocytic lymphoma, as seen hairy cell leukemias, were suggested B cell origin.
In five cases of T zone lymphoma were not shown distinct T-cell-like reaction products. It might be indicated that there is apt to decrease the esterase activities of lymphocytes in the process of neoplastic and/or blastic proliferation.

View full abstract

By using ANAE staining we report here the changes of T cells and plasma cell series in lymph nodes of 11 cases of tuberculous lymphadenitis, 9 cases of Piringer's lymphadenitis (Toxoplasmosis), 2 cases of sarcoidosis, 2 cases of syphilis and 3 cases of Morbus Pfeiffer.
In tuberculoses, toxoplasmoses and Morbus Pfeiffer T cells were prominent, particularly in Morbus Pfeiffer.
In sarcoidoses and lues both T and plasma cell series were increased.
In granuloma-forming diseases, such as tuberculosis, lues and sarcoidosis, it could be assumed that there are some different distributions of T and B cells between active granulomatous lesions and the other areas in the lymph node.
So, it should be needed that histological ANAE staining must be done.

View full abstract

We investigated the quantitative changes of T-cell and plasma cell series in the lymph node imprints of 12 cases of nonlymphatic leukemia involving lymph nodes by using ANAE-staining. In general, ANAE activities were decreased in almost cases, except two cases of CML.
In two cases of AML granular positive cells were increased. It suggested that there happened the differentiation from B-cell to plasma cell.
A case of AMoL was shown an extreme decreasing of ANAE activities of lymphocytes with these lymphoblastic changes.
Many recent reports are indicated that the various types of composite leukemias and lymphomas could be occurred with blast crisis, spontaneously and/or by drug therapy.
Our report might reflect a few findings of recent reports.

View full abstract

By using Alpha-Naphthyl-Acetate-Esterase-Staining (ANAE), we investigated the rest population of T-cell in diffue lymphomas of B-cell origin, which consist of 15 cases of CLL, 30 cases of IC, 5 cases of PRL, 17 cases of CC and 16 cases of IML. The percentages of Droplet cells changed from case to case, but the highest percentage was seen in IML (19.90 %), on the contrary, the lowest CLL (7.80%) . It may be suggested that even diffuse lympho-mas of B-cell type have relatively many T-cell Rest Populations, considering there is seen the fact of absolute number increasing of Tγ-cells in B-cell neoplasms and in addition, some parts of T-cell are ANAE negative.

View full abstract

The quantitative changes of T-cell and plasma cell series of 28 cases of metastatic tumor in the lymph node imprints were investigated by using ANAE staining. Only 4 cases in 16 metastatic carcinoma lesions were increased T-cell, on the contrary, plasma cell series were relatively preserved and in several cases rather increased. Each one case of neuroblastoma and Ewing-Tumor was revealed T-cell predominance, but its mechanism was obscure. One case of medullary thyroid carcinoma was shown a relative increasing of plasma cell series. 5 cases in 6 undifferentiated tumor lesions were decreased ANAE activities, particularly of T-cells. As mentioned above, and according to other reports, the prognosis of malignant tumor might be, in some degree, related by the function of T-cells and/or paracortical condition in regional lymph nodes. We hope to clarify the defense mechanism against the proliferation and spread of malignant tumors.

View full abstract

Quantitation of plasma cells and demonstration of cytoplasmic immunoglobulins using the unlabelled antibody peroxidase-anti-peroxidase (PAP) method was performed on paraffin-embedded sections of human femoral bone marrow (BM) obtained from 117 autopsy cases, including those with normal as well as pathologically increased or decreased serum γ-globulin (γ-G) levels. 35 individuals of control group showed, in 3μ thick sections, 3937±1699/mm² (mean±S.D.) of nucleated BM cell count (NBMCC), 39±34/mm² of plasma cell count (PLC) and 0.99±0.66% of plasma cell rate per NBMCC (PLR), that were virtually the same figures as those previously reported in the sternal and iliac bone marrow. A ratio of PLC per γ-G amount was 39.7±32.2% in control group, which was roughly equal to that of the other groups, with the exception that liver cirrhosis with normal γ-G level and multiple myeloma showed markedly higher ratio. A relatively linear correlation between PLC and γ-G amount was revealed. PLC might be more proper than PLR for the purpose of detecting quantitative abnormality of BM plasma cells. In most cases, cytoplasmic immunogloblin of BM plasma cells are predominantly composed of IgG, secondly IgA and IgM, and scarcely IgD and IgE. In light chains, κ chain was equal to or a little higher than λ chain. Abnormal, but not really monoclonal, increase in IgA, IgM, IgE, κ and λ chains were observed in some cases, but could not be accompanied with increase in serum immunoglobvlin of the same immunoglobulin classes.

View full abstract

Antigenicity of HLA antigens in human cell-mediated lympholysis (CML) was studied by using phytohemagglutinin-induced lymphoblasts (PHA-blasts) and Epstein-Barr virus-induced lymphoblastoid cell lines (EBV-LCL) as target cells. 1) In CML in families using PHA-blasts as targets, effector cells stimulated with the haplotype, which was present on the stimulator cells and absent on the responder cells in mixed lymphocyte reaction (MLR), killed the target cells from the family members possessing the haplotype. 2) Effector cells stimulated with the unrelated cells carrying different HLA-B antigen killed the third party target cells having the identical HLA-B antigen with the stimulator cells. However, the effector cells generated with the stimulation of HLA-A antigen were weakly or not cytotoxic to the targets. 3) Effector cells, generated with HLA-A, B, C-identical and DR-different combinations killed EBV-LCL targets which shared no DR antigens with stimulator cells, in addition to EBV-LCL targets with shared DR antigens. Because of this extra reaction it was not concluded that HLA-DR antigens were the real targets in CML. 4) The extra reaction might be caused by natural killer cells (NK) augmented in vitro culture, because some of the EBV-LCL used as targets had NK sensitivity to unprimed peripheral blood lymphocytes. 5) EBV-LCL possessing different HLA-D antigens, DYT and DKT2, and DR4 antigen in common were used as targets in CML to examine whether HLA-D region antigens were indeed the target antigens. Effector cells stimulated with the cells carrying DYT antigen killed the target cells with DYT antigens but not LCL with DKT2 antigen. On the other hand, effector cells stimulated with DKT2 specificity lysed the target cells with DKT2 specificity but not LCL with DYT specificity. Therefore, it was clarified HLA-D antigens are the actual target antigens in human CML.

View full abstract

Weakened Trimeresurus Flavoviridis with 70 % ethyl alcohol or other substances was intravenously administered to mice and rabbits to examine its toxic effect on the Langerhans' islands acinar cells of the pancreas, experimentally applying the method to induce specific hemorrhage mainly at the Langerhans' islands. The main target organ of hemorrohage in intravenous administration of toxoid Trimeresurus flavoviridis in this study was the lung, and most of the treated animals died of pulmonary hemorrhage, without manifestation of much effect on the pancreas either with internal or external secretion. The target organ which developed hemorrhage by the administration of weakened Trimeresurus flavoviridis with 70 % ethyl alcohol was specifically changed to the Langerhans' islands in the pancreas in stead of the lung. The amount of alcohol toxoid T, flavoviridis required to cause hemorrhage in the L. 's islands was 1.00 mg-2.00 mg per mouse weighing 22 g±2 g ; more than this level caused death, while less didn't cause hemorrhage. No hemorrhage was observed in the L.' island in the lower dosage group of 0.50 mg/mouse. However, widespread vacuolar degeneration and vacuolation were found in the acinar cells of the pancreas, and most of the vacuoles showed positive stain with amylase staining by the PAP method, suggesting that Trimeresurus flavoviridis induces hypersecretion of the pancreas in terms of immunohistology. From the result stated above, it is presumed that the pancreatic impairment model produced by the use of weakened Trimeresurus flavoviridis can be used as a model of the L. 's islands hemorrhage, if attention is paid to the hemorrhagic factors, and as modes for cell involvements with attention to the cytotoxic proteinase, and that this is of a great value in the development of experimental models of the pancreatic diseases.

View full abstract

The five major histone fractions in Ascaris lumbricoides var. seam were isolated and purified. The histone fractions were identified by polyacrylamide gel electrophoresis and by amino acid analysis. Studies on amino acid composition indicated that the histones were homologous, if not identical, to the H 1, H 2 A, H 2 B, H 3 and H 4 histones of calf thymus.The yield of histone was 5.34 mg per g of wet weight of the parasite and the value was slightly larger than for mammalian tissues. The yield of DNA was 2.50 mg per g of wet weight and, cosequently, the ratio of histone to DNA was estimated to be 2.14. The relative amounts of histone to the total histone in H 1, H 2 A, H 2 B, H 3 and H 4 fractions were 27.0, 8.8, 44.8, 5.4 and 4.0%, respectively. This finding on the remarkably low proportions of H 2 A, H 3 and H 4 and the high proportions of H1 and H 2 B of Ascaris suum appeared to be species as well as tissue specificity-dependent. The electrophoretic behaviors of each histone fraction were quite similar to those from calf thymus. Species difference was particularly noticeable in H1 fraction of Ascaris lumbricoides var. suunt, but evidence has been presented that this parasite contained five major histone fractions which were homologous to those of higher eukaryotes. The present study strongly confirms earlier observations on the conserved properties of histone proteins in nature.

View full abstract

The activities of branched-chain amino acid aminotransf erase (L-branched-chain amino acid : 2-oxoglutarate aminotransf erase : EC 2.6.1.42) and its isozymes of intestine, ovary and muscle of Ascaris lumbricoides var. suum have been studied. The activity of transaminase was found to be approximately 10-fold in the intestine than that in ovary, while muscle showed negligible activity. The best substrate among three amino acids tested, leucine, isoleucine and valine, was found to be depending on the tissue used. In the intestine and ovary, transaminase activity in the supernatant fraction was found to be about 5-fold higher than that in the mitochondrial fraction. The optimal pH of the enzyme was 8.5. 2-Mercaptoethanol was one of the most effective enzyme activators. Enzyme activity in the intestine of Ascaris corresponds to approximately one-third of that in rat pancreas and stomach, which were thought to be highest. DEAF-cellulose column chromatography of supernatant fractions from intestine, ovary and muscle revealed that ovary and muscle contained only one isozyme, enzyme I, while intestines contained two isozymes, enzyme I and II . The latter isozyme was previously reported by others to be found only in normal rat liver.

View full abstract

Pharmacodynamics of catecholamines and its' effect on T wave area of ECG were studied in healthy subjects. Noradrenaline (Nad : 0.5 mg), adrenaline (Ad: 0.4 mg), isoproterenol (Isop: 0.05 mg) and saline were injected subcutaneously. Blood pressure, heart rate, symptom and ECG were compared with those before and after drug administration. Myocardial oxygen consumption was estimated from Katz index (mean blood pressure×heart rate) and Pressure Rate Product index (PRP index : systolic blood pressure×heart rate) . Ad and Isop caused increase in systolic blood pressure and decrease in diastolic blood pressure and decrease in heart rate. Increase in Katz index and PRP index and decrease in T wave area were observed. While, Nad caused increase in systolic and diastolic blood pressure, and decrease in heart rate. Katz index and PRP index did not change and T wave area slightly increased. There were significant negative correlation between changes in PRP index and T wave area. These results suggest that decrease in T wave area by Ad and Isop is partly related to relative ischemia in endomyocardium due to increase in myocardial oxygen consumption.

View full abstract

Two cases of dermatomyositis associated with internal malignancy (I.M.) and a typical case of childhood dermatomyositis were reported. Case 1: A 65-year-old woman had a typical Gottron's sign and a purplish-red heliotrope erythema and myalgia of both upper arms and thighs. X-ray examination revealed lung cancer, which had no longer the indication for an operation. She died about 13 months after the onset of the disease. Case 2: A 49-year-old woman had a slight erythema of the face and indurated erythema of the lumbar region and stiffness of the muscles of neck and thighs. Colon cancer was found. Her skin and muscle symptomes were improved after its surgical exision. Case 3: A 8-year-old girl had an abrupt onset of muscle and skin symptomes. Steroid therapy was effective for her as it was in children's cases and her muscular and cutaneous conditions improved rapidly. The 84 cases of dermatomyositis associated with I.M, available in the Japanese dermatological literature for the recent 10 years showed that the cases whose skin symptomes preceded I.M, were 86 % and that the cases whose I.M, preceded skin symptomes were, to the contrary, 13 %. Among the associated I.M., gastric cancer was the highest (44 %) in the frequency. The time that elapsed until the skin lesion developed preceding I.M, was one month to 3 years and that until I.M, was diagnosed preceding the skin lesions was one month to 30 years. In the 110 cases of childhood dermatomyositis, skin symptome developed as the first sign in about 50 %, but the elevation of specific myogenic enzyme activity was shown in less than 50 %.

View full abstract