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<p>A combination of agarose gel electrophoresis and a newly developed technique of electro-affinity transfer was applied to the detection of circulating immune complexes of human alpha-fetoprotein (AFP) and anti-AFP. After electrophoretic transfer to nitrocellulose membrane, to which affinity-purified polyclonal horse antibodies to human AFP were bound, the membranes were treated with or without rabbit immunoglobulins to human AFP, followed by overlaying with horseradish peroxidase-labeled goat anti-rabbit IgG for color development. Artificial complexes formed in vitro from human AFP and rabbit anti-AFP were clearly separated from free AFP by the agarose electrophoresis. The complexes were stained 20-40% as dark as the equivalent amount of free AFP by treatment with rabbit anti-AFP, and 10-20% as dark without the antibody treatment over a wide range of antigen-antibody ratios.</p>

Transaminative metabolism of L-cysteine was investigated using homogenates of guinea pig liver and kidney. L-Cysteine was transaminated in the presence of 2-oxoglutarate and the homogenate of either liver or kidney. S-(2-Hydroxy-2-carboxyethylthio)cysteine (HCETC) (3-mercaptolactate-cysteine disulfide) was formed by liver homogenate, but the amount was very small. On the other hand, a relatively large amount of HCETC was formed in the presence of kidney homogenate. Transamination between 3-mercaptopyruvate and certain amino acids was catalyzed actively by both liver and kidney homogenates in the presence of L-glutamate. However, more half-cysteine was formed by liver than kidney, and more HCETC was produced by kidney than liver. L-Glutamate was the most potent amino donor, and L-aspartate strongly inhibited the reaction. Results indicate that L-cysteine can be transaminated both in liver and kidney of the guinea pig, and that kidney is more active than liver. 2-Oxoglutarate is the most active 2-oxo acid for cysteine transamination. Oxaloacetate (and aspartate in the reverse reaction) is inhibitory to the reaction. These results are in agreement with the previous conclusion that cysteine aminotransferase is identical with aspartate aminotransferase.

1. Absorption maxima of hydrochloric biliverdins derived from the natural indirect bilirubin existed at 680 mμ and 375 mμ, but the maxima of biliverdins purified on the column of silica gel existed at 640 mμ and 390 mμ. 2. The natural salt-form bilirubin was oxidized by hydrochloric acid to biliverdin, of which absorption maxima existed at 685 mμ and 370 mμ in a methanolic solution as well as in 5% hydrochloric methanol, but the purified biliverdin in chloroform solution showed the maxima at 640 mμ and 390 mμ. 3. The natural ester-form bilirubin could be transformed into biliverdin by oxidation of its alcoholic solution in the presence of hydrochloric acid. The crude biliverdin had absorption maxima at 645 to 655 mμ, 600 mμ and 320 mμ, and the crude hydrochloric biliverdin had the maxima at 665 to 675 mμ, 620 mμ and in the near ultra-violet range, while the purified biliverdin in chloroform solution had the maxima at 640 mμ and 380 mμ. 4. The biliverdins derived from the indirect, salt-form and ester-form bilirubin had quite similar absorption maxima after purifications by adsorption chromatography.

Diacetyl can serve as an acetyl donor for the forlnation of citrate and the acetylation of sulfanilamide in the dog heart homogenate. Diphosphothiamine and coenzyme A are essential for these reactions.

Separation of both forms of the direct bilirubin were carried out from the dog's gallbladder bile, and further isolations of them were also done. 1. The natural salt-form bilirubin was isolated after separation on the column of aluminium oxide with a n-propanolic aqueous solution. 2. The natural salt-form bilirubin was obtained in amorphous yellow powders which were strongly hygroscopic and easily soluble in water and methanol but not in chloroform or carbon tetrachloride. An aqueous solution of these powders showed both the direct diazo and Gmelin reaction, but neither Ehrlich's aldehyde nor Schlesinger reaction. The salt-form bilirubin was transferred into chloroform only when some quantities of hydrochloric acid were added to a mixture of chloroform and an aqueous solution of it. 3. The absorption maxima of the natural salt-form bilirubin existed at 420 to 430 mμ in a methanolic solution and at 425 or 435mμ in 50% or 10% n-propanol. 4. The natural ester-form bilirubin was isolated after separating on the column of silica gel with a chloroformethanolic mixture. 5. The natural ester-form bilirubin was obtained in amorphous greenish yellow powders. It was further hygroscopic and easily soluble in water and methanol but not in chloroform or carbon tetrachloride. An aqueous solution of it showed the direct diazo and Gmelin reaction, but neither Ehrlich's aldehyde nor Schlesinger's reaction. No pigment was transferred into chloroform even if some quantities of hydrochloric acid were added to a mixture of chloroform and an aqueous solution of it, but did by saponification with 5% methanolic potash. 6. The absorption maxima of the natural ester-form bilirubin existed at 415 mμ in both methanolic and aqueous solutions.

Descriptions are carried on the method how to separate the indirect bilirubin from the chloroform extracts of the dried dog's gallbeadder bile by adsorption chromatography. 1. The optimal concentrations of the bilirubin content were 2 to 4 mg/100 ml when 1 ml of the sample was adsorbed on the Tswett tube of about 10 mm diameter. 2. Though several zones of the indirect bilirubin were separated on the column of silica gel when developed with various solvents, these zones were proved to be mingled with some oxidized or other intermediate products and the separation like this was thought to owe to the activity of the adsorbents. 3. The chromatogram of the crystalline bilirubin resembled to the one formed by the indirect bilirubin in the chloroform extracts. 4. The chromatogram of the chromatographically separated indirect bilirubin was similar to the former. 5. The absorption maxima of a chloroform solution of the natural indirect bilirubin existed at 450 mμ in the visible range, and it was the same as the maxima of the crystalline bilirubins.

<P>The effect of P³² in the experimental Japanese B Encephalitis infection was investigated. A remarkable delay of infection was observed, when a therapeutic dose of P³² was administered intraperitoneally to the mouse which had been inoculated intracerebrally with the virus suspension at the concentration of about LD50. Almost equal results were obtained by intraperitoneal administrations to the mice when they had deen inoculated intravenously or intraperitoneally.

1. Sinomenine and Irgapyrin, the two antirheumatics known to be capable of releasing histamine, caused a marked gastric secretion in the unanesthetized dog. 2. The facial edema and itching associated with histamine release by sinomenine was almost completely eliminated by NeoAntergan, but the gastric secretion was not suppressed, or rather increased - an observation also reported by Paton and Schachter with Compound 48/80. This indicates that the histamine release cannot be markedly prevented by antihistamine agents in this animal. 3. The gastric secretion induced by Irgapyrin was not suppressed by Neo-Antergan but Irgapyrin originally never caused other symptoms associated with histamine release. This is probably due to the antihistamine action inherent in this compound itself. 4. No such histamine-releasing activity, as determined by gastric secretion, could be observed in aminopyrine or butazolidine sodium, the components of Irgapyrin. 5. Sinomenine, differing from Irgapyrin and Compound 48/80, was ineffective by intramuscular injection.

Our methods of tissue culture of the bone marrow, lymphnodes, and peripheral blood were described. Furthermore, for the purpose of promoting wide clinical application of bone marrow tissue culture, our simple vital inspection method was also stated.

In decerebrated cats the impulse discharges were detected by means of an unipolar microelectrode from a single inspiratory neuron in the respiratory centers, and the change in discharge produced by administration of stimulating, as well as depressant agents, was studied. The results were summarized as follows. 1) Inhalation of the air containing all. excess of carbon dioxide, as well as the air deficient of oxygen and the intravenous administration of a small dose of caffeine, aminocordine and lobeline produced a) a remarkable increase of frequency of impulses in the stage of constant frequency of the volley, b) a rapid increase of frequency of impulses in the stage of crescent frequency of the volley and c) a shortening of the duration of the volleys, as well as of silent periods. 2) In narcosis by a moderate dose of morphine, as well as in the recovery stage from apnea produced by over-ventilation, there were observed the phenomena which were exactly opposite to those described in 1). 3) It was concluded that a most essential sign by which one can discern whether the activity of the respiratory Genters is raised or depressed, is the changes of the frequency of impulses produced from an inspiratory neuron. The expense of this research was defrayed from the grant in aid of the Ministry of Education.

1) As the boundary of the outgrowth zone of the bone marrow culture is sharply defined from the surrounding area and the cellular density becomes very high in leukemia, diagnosis can be made with ease and certainty. 2) The differential diagnosis between aleukemic leukemia and aplastic anemia becomes easy by application of the tissue cuIture method. 3) The vital observation of blood cells were used in classification of leukemia by cellular systems, and it was discovered that there were a lot of patients with monocytic leukemia and relatively many with acute lymphatic leukemia. 4) Because the cellular growth in the bone marrow culture of aplastic anemia declines extremely and abundant fat cells are observed, the diagnosis of this disease is easily made by employment of this method.

1. A. simple hot-plate apparatus, with comparatively good accuracy in the control of temperature, chiefly composed of a circular lead plate embedded with nicrome-wire heater was devised in order to improve the complications and inconveniences of the hot-plate apparatus used to date in algesimetric determination with mice. 2. The reaction times measured with this hot-plate maintained at 55°C with 500 normal mice showed an average of 9.96 ± 1.58 seconds and about 90% of the total mice exhibited reaction time of 7-13 seconds. With animals showing reaction time within this range, the daily mean reaction time did not vary with measurement once a day for consecutive days and the effect of drugs with comparatively weak analgesic effect was well reproduced with small number examples, such as 12 mice to a group.

The idiopathic thrombocytopenic purpura was described by Werlhof as an independent disease first in 1738. Kaznelson reported the excellent effect of splenectomy for its chronic type in 1915. For the genesis of its thrombocytopenia, there have been many theories to be concluded into the followings, 1) the development of an auto-immune mechanism resulting in platelet destruction, 2) increased platelet destruction in the spleen, 3) the inhibition of platelet production from the marrow megakaryocytes by a humoral factor produced in the spleen, 4) both increased destruction and decreased production of the platelet. Among the above four theories, the third one is the most popular in the chronic type.

1) When guinea pig serum was warmed at about 65°C, part of its protein became denatured resulting in the formation of a colloidal component C, which has approximately the same electrophoretic mobility as that of α-globulin. 2) The electrophoretic examination shows that the serum heated at 65°C after adding the glucose is devoid of colloidal component C. 3) The inhibitory action of sugar and polyatomic alcohol on the heat inactivation of complement is galactose> saccharose > glucose> sorbit > mannit > glycocoll> glycerin in descending order. The inhibitory action of sugar and poly atomic alcohol on molecular aggregation by heating is of the same order as complement activity.

1) The fate and rate of degradation of I131 labelled rabbit γ-globulin, which retained its native antigenicity and antibody specificity was studied in the guinea-pigs. 2) Blood elimination rate of heterologous γ-globulin is higher than that of homologous γ-globulin. 3) Denatured and digested γ-globulin departs from the blood more rapidly than the native one, and urinary excretion rates of denaturated and digested γ-globulin are higher than that of the native one. It is inferred, therefore, that the denatured and digested γ-globnlin is more liable to be resolved and decomposed in the reticulo-endothelial organs than the native one. And the value obtained from the urinary excretion reflects the rate of protein break down in some cellular compartments. 4) Following the plasmaphresis the increase in antigen elimination was lessened and delayed as compared with control animals. 5) The organ distribution of heterologous I131-γ-globulin is to the lymphnode> the spleen> the liver> the lung> the kidney> the intestine in descending order. Heterologous I 131 -γ- globulin is deposited in greater quantity in the reticulo-enclotherial organ than other single organ. 6) Following the intravenous injection of I131 labelled antigens, the ratio of the specific activity of mitochondria and microsom to that of whole liver homogenate was determined over a period from 15 minutes to 3 hours in guinea-pigs, and following results were obtained. a) Organ and intracellular distribution of Il3l labelled homologous γ-globulin shows no great difference compared to that of heterologous one. b) The intracellular distribution of heterologous γ-globulin is in mitochondrial> microsomal> nuclear fraction in descending order. c) The heterologous γ-globulin quantity of mitochondrial fraction or microsomal fraction in the spleen is higher than that of the liver. 7) The antibody distribution of intracellular glanules measured in terms of radioactivity with a Geiger-Muller counter, after the reaction of I131 labelled antigen. The quantity of distribution of intracellular glanules decreases in mitochondrial fraction> microsomal fraction> nuclear fraction in descending order.

HST virus which was isolated in 1950 from the roshida ascites tumor by Hamazaki and his associates is a pantropic virus which creates a unique inflammatory granulation in mice. When virus of an acute infections disease was inoculated on embryonated eggs, not only the egg membrane but also the chick embryo were infected more or less, and when the number of virus increased the chick embryo died, terminating the development of the egg. However, the tumor inducing virus which represents the Rous virus does not cause heavy disturbances in the embryo and it is well known that chick hatched from this egg can long maintain health unless it is subjected to a provocative factor. HST virus is no exception to this example and though it is inoculated on an embryonated egg it does not cause any serious disturbance on the embryo. The tissue changes of the chorio allantois infected by the "Virus were the focal proliferation and necrosis of ectodermal epithelium, the proliferation of the mesenchymal cells of the mesodermal layer adjacent to these foci, accompaning infiltration of lymphoid cells and leukocytes with edema, especially eosinophilic leukocytes. By these tissue changes a terrace-shaped thickening of the membrane was the result. In the viscera of the chick embryo a special change in the liver was seen, i. e., along the edge of the liver greyish white nodules submacroscopic to miliary in size appeared. The principal pathologic change of the foci is the coagulation necrosis of the liver parenchyma and only a slight infiltration about the periphery of the foci was observed. Moreover, proliferation of mesenchymal cells occurred next to the walls of the large blood vessels of the liver (principally, the portal veins) and with the added infiltration of a small number of lymphoid cells and leukocytes sharply defined nodular foci were formed. Though this was a rare instnace, similar pathologic changes were seen also in the walls of the blood vessels of the cerebrum stem of the embryo and along the periphery local gliosis was observed.

Histopathological investigations were carried out on five fatal cases of a type of polyneuritis of unknown etiology diagnosed as Landry-Guillain-Barre syndrome, which endemically occurred in children in the regions surrounding the Inland Sea of Japan. The most characteristic pathologic feature in the nervous system was pronounced patchy degenerative changes with slight or moderate degree of inflammatory cell response of focal type in the peripheral and cranial nerves, predominantly in the nerve fibers of the spinal and cranial roots. In the spinal cord, medulla, pons, and in some portions of the cerebrum and cerebellum, engorgement of the small blood vessels as well as edema and the less predominant scattered degenerative changes of ganglioncells and nerve fibers with extremely slight degree of glial response and sparse perivascular cell collections were encountered. The cerebrospinal meninges displayed edema and congestion of the pial blood vessels with focal collections of a small number of lymphocytes and/or monocytes. No advanced involvement of the anterior horn of the spinal cord in a strict sense of anterior poliomyelitis was, however, recognized. These changes may lead the histopathologic diagnosis of the present disease to infectious encephalomyelo-polyradiculoneuritis or a type of infectious polyneuritis. The main histopathologic changes in the visceral organs were a moderate degree of engorgement of the small blood vessels, degeneration of parencymatous organs such as the liver and kidney, hyperplasia or follicular atrophy of the lymphatic tissues, interalveolar pneumonia, focal myositis, and slight degree of round cell infiltrations in the interstitial tissues of the other viscera, such as the liver, heart, and gastrointestinal canal. Based upon the observations on the histopathological changes as well as clinical manifestations, discussions were made on the pathogenesis and etiologic factor of the present endemic disease with critique on the literatures.

For the purpose of obtaining the dibasic acid indirect bilirubin in a pure state from the dried canine cholecystic bile, an optimal developing solvent was selected by paper partition chromatography as a preliminary experiment, and it was isolated on cellulose column as an applied experiment. 1. The dibasic acid indirect bilirubin was separable at the starting point in a pure state by paper chromatography under development with the top layer of a n-butanol, acetic acid, water mixture (4:1:5). 2. The dibasic acid indirect bilirubin formed a fixed band at the upper starting place on cellulose column under development with the top layer of a n-butanol, acetic acid, water mixture (4:1:5), and no other substance could be detected there. 3. The dibasic acid indirect bilirubin existing in the fixed band could be eluted out into chloroform with a 1% acetic acid solution. An orange yellow powder was obtained from the eluate by evaporating the solvent in vacuo. 4. Thus separated orange yellow powder agreed well with the crystalline bilirubin in the solubility into organic or inorganic solvents and in the spectrochemical characteristics as well as in the chemical properties.

Blood cells of schizophrenics differ in many points from those of normal subjects. First of all the shape of them is flat and thin. This tendency is more marked in old group than in new group; the volume is small; flat corpuscles are more numerous in them in the normal; and the blood resistance against diluted saline solution is stronger than that of the normal. It has long since been known that the rate of corpuscle sedimentation is being accelerated in schizophrenics. A simple physical cause that blood corpuscles are flat and numerous can explain this phenomenon. It is said that there is an antisphering substance among the factors controlling the thickness and roundness of blood corpuscles. Yet it has not been determined whether this substance on the surface of the blood of schizophrenics is large or small. Blood corpuscles are said to lose their peculiar disc-shape and to be completely destroyed at the pH of 9.2 when the antisphering substance is removed from the surface of blood corpuscles. The lower the pH is the better is the absorption of this substance on the surface of blood corpuscles; and it seems that the more this substance attaches itself to blood corpuscles the greater is the degree of flatness and in this connection it is interesting to note that the pH of schizophrenic blood is low in low in general. On the other hand, however, sphericity is increased at the time when the acidity of blood is increased due to a sudden movement of acidic substances immediately after ECT. Again in the case of coma of insulin treatment, blood tends to be alkaline and even then an increase in the sphericity of corpuscles is indicated. Consequently it seems tnat the roundness of blood corpuscles is not solely dependent upon antisphering substance and pH.