In order to clarify the leukemogenesis by x-ray, observations were carried out on the peripheral blood pictures and on the histological changes of hematopoietic organs from time of X-irradiation to the onset of leukemia with lapse of time, and in addition, the developmental course of leukemia was pursued by Hiraki's clinical tissue cultures. As the result the following conclusions were drawn. 1. After one whole body irradiation of 350r on the RF strain of mice, leukemia developed in 77.3% of them. Out of them 75.4% proved to be lymphocytic leukemia while 1.8% myelogenous leukemia. In the former group some were of the thymoma type that presented swelling of the thymus and the others of the non-thymoma type without such swelling of the thymus. 2. After the X-irradiation, these mice showed hypoplasia of hematopoietic organs, which was followed by reactive proliferation, and in those of the thymoma type leukemic changes originated in the thymus whereas in those of the non-thymoma type the changes commenced in the spleen or in lymph node. 3. One month before the onset of leukemia, that is, in the so-called preleukemic stage, there could be observed a decrease in the leukocyte counts and an increase in the eosinophil counts, and in addition, the splean, lymph nodes or thymus showed such findings what might be histologically considered as leukemia, but by the clinical tissue cultures the findings rather suggested that it was in the stage showing hypoplasia. From these findings it has been concluded that leukemia induced by X-ray irradiation originates in the thymus in the case of lymphocytic leukemia of the thymoma type, while in the case of lympocytic leukemia of the non-thymoma type leukemia makes its appearance first in the spleen or in lymph nodes after passing a hypoplastic stage.
For the elucidation of leukemogenesis of mouse leukemia induced by X-ray, a study was carried out on various factors influencing the development of leukemia. Namely, an effort was made to see the relationship between such factors as the age of mice receiving X-irradiation, the dose and the method of irradiation, the strain and sex of mice on one hand and the rate of leukemia induction on the other. Also a comparative study was made of the types of mouse leukemia elicited by X-ray. The results of the study are stated as follows. 1. In mice of the RF strain given one whole body X-irradiation of 350 r, and in both C57BL and C3Hf mice given four whole body irradiations of 160 r each at the interval of 4 days to the total dosage of 600 r, the rate of leukemia induction was 77.3%, 88%, and 20% in the respective groups. 2. It has been demonstrated that the rate of leukemia induction was highest in mice of the RF strain 30-40 days old, and this rate decreased along with advancement in the age (several months old) of the mice. 3. As for the rate of leukemia induction in mice of the RF strain 30-40 days old, that received one whole body X-irradiation of 350 r, 22 (78.5%) out of 28 male mice developed leukemia while 19 (73.0%) out of 26 female mice, developed leukemia, showing no significant difference in sex. 4. It has been found that the rate of leukemia induction in the RF strain mice of 30-40 days old after one whole body X-irradiation increases with the dose of the irradiation, reaching its maximum at the dose of 350 r, but with the dose of 450 r the rate of leukemia induction rather decreases. 5. Of lymphocytic leukemia induced in the RF strain mice by X-irradiation, there were two types, that is, thymoma type and non-thymoma type, and there could be observed distinct differences in the peripheral blood pictures as well as in the patterns of cell infiltration into the liver between these two types. In addition, some of these lymphocytic leukemic mice presented a neutrophilic leukemoid reaction. There was only one mouse that developed myelogenous leukemia.
By means of phase-contrast microscopy, vital staining, and fluorescence culture method, cytological and tissue culture observations were carried out with the leukemic cells of RF strain mice after X-irradiation. As the result the characteristics of lymphoblasts and myeloblasts were confirmed and also from the growth patterns of the cells from various organs of the leukemic mice it was possible to differentiate the two classifications of the thymoma type and the non-thymoma type of lymphocytic leukemias as already described in a previous report (Part 2). The results of this study support the view of the author on the onset mechanism of leukemia induced in RF mice by X-irradiation; namely, in the case of thymoma type lymphocytic leukemia leukemic changes originate in the thymus, whereas in non-thymoma type lymphocytic leukemia the leukemic changes originate in the spleen or in lymph nodes.
As one of the series of studies on radiation leukemogenesis, changes in the peripheral blood picture occurring with lapse of time were pursued with the Cb, D103, StA, and Zb strains of mice (30-50 days old) after a single whole body X-irradiation of 350 r. The results are summarized as follows. 1. Both erythrocyte counts and hemoglobin contents showed the minimum values 2 weeks after the irradiation, and after recovering once they tended to decrease again. Reticulocytes decreased to about 2-4‰ immediately after the irradiation but by the second week they increased to about twice the number in normal condition, and thereafter they returned to the normal count. This change was most marked in the D103 strain, but ultimately the changes in the erythrocyte series presented similar curves in all the four strains mentioned above, and these changes did not differ from those observed in RF strain. 2. The number of leukocytes decressed rapidly to as low as 2, 000 immediately after the X-irradiation. The susceptibility to X-ray irradiation was higher in the leukocyte series than in the erythrocyte series. 3. Lymphopoiesis was more sensitive to X-irradiation than myelogenous hematopoiesis, and the decrease of lymphocytes was greater than that of neutrophils, showing a relative neutrophilia for several weeks after the irradiation. 4. After once recovering, the leukocyte counts in the Cb, D103, StA, and Zb strains of mice all did not show such a transient decrease as observed in the preleukemic stage of RF strain about 13 weeks after the irradiation. In addition, lymphocytic leukemia did not develop. 5, In the Cb, D103, StA, and Zb strains irradiated with X-ray there could be recognized no severe eosinophilia such as observed in the RF strain that received X-irradiation. From these findings it has been confirmed that there can be recognized no marked differences in the peripheral blood picture immediately after a single whole body X-irradiation of 350 r between the RF strain of mice that develop leukemia by X-ray, and the Cb, D103, StA. and Zb strains that do not develop leukemia by X-irradiation, but in the observations conducted thereafter along with lapse of time there can be seen several distinct differences in the changes of the leukocyte series between them.
In the series of studies being conducted on radiation leukemogenesis, observations lasting for one year were carried out to see the changes in the hematopoietic organs with lapse of time, using 30-50 days old mice of the Cb, D103, StA, and Zb strians after a single whole body X-irradiation of 350 r. The findings of the study are presented in the following. 1. In the observations lasting for one year, there was no development of leukemia in 18 mice of the D103 strain, in 19 mice of the StA strain, and in 13 mice of the Zb strain. In one out of 20 mice of the Cb strain, however, leukemia did develop, and it was confirmed to be myelogenous leukemia from its peripheral blood picture and tissue culture. Cell transplantations from this mouse to other mice of the same strain were not successful. The changes in hematopoietic organs occurring with lapse of time after X-irradiation in aech strain of mice that did not develop leukemia are as described as follows. 2. Lymph nodes, spleen and thymus after the irradiation showed degeneration, necrosis and lymphoctyopenia, with a decrease in their weight. 3. Two weeks after the irradiation the spleen was transiently enlarged, showing an acceleration of splenic function. This was striking in mice of the D103 strain. 4. After the decrease in the weight of lymph nodes, spleen and thymus, the proliferation of reticuloendothelial cells and lymphocytes was accelerated, but this tendency in any organs was limited to the proliferatioan of mature cells only. There could also be recognized distortion of thymus in some. 5. The changes in the bone marrow were slight and the peripheral blcod showed a relative neutrophilia. 6. These changes were similar in each strain of mice, especially those changes in the initial stage were exactly identical with those observed in RF strain mice. In the RF strain mice, however, young cells proliferated at a relatively early stage whereas in the resistant strains such as the Cb, D103, StA and Zb strains the reactive proliferation of only mature cells could be observed. 7. Liver cells were greatly affected by X-ray, showing degeneration, necrosis, and vacuolization even 16 weeks after the irradiation, but no specific infiltration of leukemic cells as in the case of mice of the RF strain could be demonstrated. From these findings it is assumed that mice of the RF strain possess leukemogenic factor in the form of provirus and this is activated by X-irradiation, while mice of the Cb, D103, StA or Zb strains do not have provirus or has provirus in a state in which it is difficult to be activated by X-irradiation.
With the purpose of studying a possibility of the so-called “two-stage mechanism” in radiation leukemogenesis, a series of studies was conducted to compare the rate of leukemia development, using RF strain mice divided into a set of groups of mice that received one whole body X-irradiation of 150 r, 250 r, 350 r, respectively, and another set of groups similarly irradiated with respective doses and repeatedly painted with 5 % croton oil in benzene solution after irradiation. 1. As a result it has been found that in the group receiving 350 r there was no significant difference in the rate of leukemia development between the group receiving the irradiation only and the group that was painted with croton oil solution in addition to the irradiation. 2. In the group that received 250 r, in the group that received croton oil painting immediately after the irradiation or one week after the irradition, there was observed a distinct increase in the rate of leukemia development, while there was no difference between the group that received croton oil painting before the irradiation and the group that received the X-irradiation only. 3. In the groups that received 150 r of the irradiation, leukemia did not develop but it developed only when croton oil was concurrently painted. This finding suggests that leukemia does not develop even when provirus is activated by X-irradiation, if the promoting agent, croton oil is not administered thereafter. 4. In the case when both X-irradiation and croton oil painting were given there could be recngnized leukemias of the so-called thymic type and the non-thymic type in about the same number. 5. In the group given one whoie body irradiation of 250 r and 100 mg of urethane in five separate doses, leukemia developed at a high rate, but in the group given urethane injection alone one of 8 mice developed leukemia. From these results it seems that the “two-stage mechanism” of cancer development as advocated by Berenblum is applicable to the radiation leukemogenesis, and it is thought that X-irradiation acts as an initiator and croton oil as its promotor. In addition, it is presumed that urethane as an initiator has ability to induce leukemia.
Clinlcal significances of the anti-lung antibody were studied on the course of the 287 cases of lung tuberuculosis patients. The results summarized as follows; 1. The sera of the patients with various kind of lung diseases were tested for anti-lung-antibody activity against human lung antigen, using the complement fixation reaction. The cases with the positive anti-lung-antibody were in 2 cases out of 4 cases of bronchoectasis, in 2 cases of 4 cases of lung abscess, in 9 cases of thoracic empyema, in 126 cases of 287 cases of lungc tuberuclosis. From the correlation between clinical types (Gakuken's classification in Japan) and anti-lung antibody the incidence of the positive anti-lung-antibody was as the flloowing ordet. A>B>F>C>D A: Exsudative Form B: Infiltrating-caseous Form C: Fibrose-caseous Form D: Fibrose Form E: Disseminated For F: Serious Mixed Form 2. In the positive anti-lung-antibody cases with lung tuberuculosis the cases with the positive anti-luny-antibody increased in titer gradually and the cases with the continuous or occassional positive anti-lung-antibody for a long period became week and the clinical signs worsened. On the other hand, the cases with the anti-lung-antibody decreased gradually in titer greatly improved. 3. On the course of the lung tuberuculosis circulating antilung-antibody increased in titer in parallel with the increased titer of complete tuberuculin antibody in peripheral blood. 4. After administration of anti-tuberculosis drugs and/or after surgical treatments in the cases with tuberuculosis of the lung the circulating anti-lung-antibody decreased in titer and was not detected in peripheral blood. 5. In the cases with tuberuculoma or lung cavity, the more the capsules of tuberuculoma or the walls of lung cavity were thin, the more the positive anti-lung-antibody cases increased in number, 6. From the distribution of the anti-lung-antibody or complete tuberuculin antibody in the lung of human tuberuculosis, it was found that the amount of complete tuberuculin antibody relatively increased in the focus of tuberuculous lesions supirior to that of anti-lung-antibody, but in the peripheral aroa of the lung tuberuculosis the amount of anti-lung-antibody was detected in higher titer that of complete tuberuculin antibody.
This attempt was made to examine the anti-lung antidody production in the experimental tuberuculosis of the lung in rabbits. The results summarized as follows; 1. Anti-lung-antidobodies were increased in titer in parallel with the development of the experimental lung tuberuculosis lesions. 2. In the experimental lung cavity formations rabbits were inoclated intracutaneously with human type killed tuberuculous bacillus (H37RV), ovalbumin and homologous lung emulsion and 1 week after inoculations with above described antigens, the injection was made aseptically into the lung of sensitized rabbits with the emulsion of 4mg human type killed tuberuculous bacillus suspended 1.0 ml incomplete djuvant. There were no significant differences in histological findings of the lung lesions among these experimentalanimals. 3. The close relationship between anti-lung-antibody and complete tuberuculin antibody was seen, and was similar to that ofhuman lung tuberuculosis patients.
Despite the fact that the pathologic changes of the foot in rheumatoid arthritis are quite specific and common, studies on statistics and pathomechanism of rheumatoid foot are relatively rare. In view of this, clinical and X-ray studies as well as study on pathomechanism of deformation of foot were carried out with 103 patients of rheumatoid foot (182 foot) that visited the outpatient ward of Orthopaedic Surgery of Okayama University for the period covering from January 1963 to March 1964. The results are in the following. It has been found that the rheumatoid patients having pathologic changes of the foot amounted to 56%, showing the incidence to be higher than what is generally thought. It is to be noted that the sites where pathologic change of ankle joint proved to be most severe are the anterior and posterior regions of medial and lateral malleolus, followed by Chopart's joint, and these proved to be the main factors for talipes planovalgus. Rep esentative deformations of the rheumatoid foot are eversion of heel, talipes planus, hallux valgus, and hammer toe. The onset mechanism of these deformations may be explained as to start with inflammation of synovial membrane due to rheumatism, which is followed by the weakening of ligaments and muscles constituting the joint capsule and the joint, then the subsequent accumulation of static and kinesiologic factor makes the pathologic deformation definitive. Differences between rheumatoid hand and rheumatoid foot are not only limited to local inflammation and anatomical elements, but also to the foot there arises an additional problem of weight bearing. With respect to functional disturbances in the case of a rheumatoid foot main complaints are less in spite of the injury of bone and deformation. when the comparison is made between Europeans and Americans on one hand and Japanese on the other, differences in the modes of living have a telling influence, and of Japanese people with rheumatoid feet they may experience some inconvenience in their routines of life but they are not so much bothered by their functional disturbance.
Increasing opportunities of applying heart-lung machine for open heart surgery, demands reducing an amount of whole blood by the addition of priming substituent. Among hemodiluents which have been studied, low molecular weight dextran (LMD) was reported to have a flow-improving effect in microcirculation by preventing or inhibiting the intravascular aggregation of red blood cells. To establish some theoretical support for this advantageous hemodilution by LMD, the author has performed clinical and experimental studies, particularly investigating mechanism of antisludging effect of LMD, and the result was summarised as follows. In hemodilution by LMD, uniform increase in electric negativity of both red blood cell and platelet was consistently observed in vitro, as well as improvement of suspension stability of blood. However, addition of each of glucose, polyvinylpyrrolidon, mannit, sorbit and amino acid to the whole blood as a diluent failed to cause any significant change in electric charge of blood cell, while improvement of suspension stability was obtained. In extracorporeal perfusion with whole blood as the priming vehicle, reduction of negative charge of blood cell was occurred. The increased negativity of blood cell in the presence of LMD was gradually decreased during perfusion, but it was kept higher than that before LMD was added, and this gave apparent benefit to the clinical extracorporeal circulation. LMD also decreased and normalized a positive charge in injured intima of blood vessel, which may probably attract negatively charged blood corpuscles, and this effect of LMD may prevent subsequent development of intravascular thrombosis. Addition of LMD to the whole blood has prevented both the decrease in electric negatitivy of blood cell and the increase in viscosity of blood and plasma, which were always observed during hypothermic procedures. Surveying the biochemical character of polimer, disadvantage in reduction of electrical negativity of the blood cell by addition of either polybrene or protamine was discussed, and possible future improvement of heparin neutralization was anticipated.
The nature of platelet clump formation following massive preserved blood transfusion was studied clinically and experimentally. A heparin-like substance was increased in the plasma of recipients following massive preserved blood transfusion, which seemed to be a nature of the platelet clump formation. The platelet clump formation is a transient reversible change, and not due to antigen-anti-body reaction. It was presumed that the decrease of platelet count in recipient's peripheral blood was caused by sequestration of platelet mostly in liver, spleen, lung and bone marrow.
The protective effects of Cysteine and MEA against X-irradiation were investigated, comparing the population of inoculated L-strain cells of the treated groups with that of the irradiated control groups on 2 days and 4 days after 800 r or 400 r irradiation. The results were as follows. 1) The protective effect of 10-4M Cysteine was observed significantly on 2 days after 800 r irradiation. 2) Cysteine of 10-5M was significantly effective on 2 days and 4 days after 800 r irradiation. 3) MEA seemed to be more effective than Cysteine (not significant).
The protective effects of AET, Cystamine and HCT against irradiation were studied, comparing the cell population of the treated groups with that of the irradiated control groups on 2 days and 4 days after 400 r or 800 r irradiation. The results were as follows. 1) The protective effect of 10-4M AET was observed significantly in both 800 r and 400 r irradiated groups. 2) The protective effect of 10-3M AET was observed on 2 days and 4 days after irradiation. 3) The protective effects were not observed in Cystamine and HCT treated groups.
The abnormal filament formation of Sal. typhi was observed on penicillin containing nutrient agar in 1u/ml. Characters of the filament cells were compared with normally harvested cells in the view points of oxidative activities to some substrates, acid agglutinating property, glucosamine content and serological properties. No difference was observed about the oxidative activities, the glucosamine content and the serological properties. With respect to the acid agglutinating property, as comparaed with the normal cells, the filament cells exhibited a property to agglutinate in broader PH range.
1. The abnormally elongated filament cell formafion was observed on Mitomycin C containing agar in 1 gamma per ml. In the filment cells, by cell wall staining method, no septum formation was observed notwithstanding cell wall structure was retained. By a method of chromatinic body staining, regularly arranged abundant chromatinic bodies were observed. 2. By a method of manometric technique, the filament cells exhibited low oxidation activities to the substrates on TCA cycle. 3. With respect to acid agglutination test, the filament cells were agglutinated at the different range as compared with normal cells. 4. In the course of the filament formation, RNA synthesis proceeded without inhibition but DNA synthesis was slightly inhibited.
Assays of the β-glucuronidase activity in sera and in liver tissues of patients with liver diseases were carried out by the method of Masuya and Tsukamoto, using p-nitrophenyl-β-D-Glucopyranosiduronic acid as substrate. The results were as follows: 1. The mean enzyme activity in normal sera was 885 units. The average activity for men was rather higher than that for women. 2. Serum β-glucuronidase activity showed a distinct increase in acute hepatitis, compensated liver cirrhosis and hepatic carcinoma and a slight elevation in recurrent chronic hepatitis, active chronic hepatitis, precirrhosis and obstructive jaundice. On the contrary, the activity was almost normal in inactive chronic hepatitis and abnormally low in decompensated cirrhosis. 3. In acute hepatitis, serum β-glucuronidase activity was markedly high during the acute phase and returned gradually and simultaneously with transaminase to the normal level as a sign of recovery. In chronic hepatitis, precirrhosis and cirrhosis of the liver, serum β-glucuronidase activity fluctuated independently of transaminases and the period of low β-glucuronidase activities was in accordance with that of hepatic failure. A rise of the activity following this period was an early sign of recovery and a return to the normal level coincided with a further clinical improvement. 4. β-Glucuronidase in liver tissues of patients with hepatic diseases showed not only an increase but also a decrease of the activity. The activity alterations were closely related to degeneration, necrosis and regeneration of hepatic cells, while cases with severe histological changes exhibited a reduced activity. 5. There was a close correlation between the activity changes of β-glucuronidase in serum and those in liver tissues. 6. These data suggest that in liver diseases the alterations of serum β-glucuronidase activity reflect sensitively active histological changes of the liver including cellular damage and repair.
Assays of the N-acetyl-β-glucosaminidase activity in sera and in liver tissues of patients with hepatic disorders were carried out by the method of Walker, using p-nitrophenyl-N-acetyl-β-glucosaminide as substrate. The results were as follows: 1. The normal activity of serum N-acetyl-β-glucosaminidase was 5.7 units in average. There was no significant difference in sex. 2. Serum N-acetyl-β-glucosaminidase activity showed a distinct increase in acute hepatitis, recurrent chronic hepatitis, precirrhosis and active cirrhosis. In active and inactive chronic hepatitis and inactive cirrhosis, the activity was slightly elevated. 3. In liver diseases the activity of serum N-acetyl-β-glucosaminidase changed independently of that of serum β-glucuronidase. 4. N-acetyl-β-glucosaminidase activity in serum showed a close correlation with that in liver tissues and such histological findings as “degeneration of liver cells”, “Kupffer cell activation”, “cell infiltration”, “proliferation of connective tissue” in the liver. 5. The elevation of serum N-acetyl-β-glucosaminidase activity has a close relation to the proliferation of connective tissue in liver diseases, while it has a close relation to the acute destructive change such as degeneration and necrosis of the hepatic cells. 6. In obstructive jaundice and hepatic carcinoma, remarkably elevated activities of serum N-acetyl-β-glucosaminidase were observed.