Okayama University Medical School

1. Mitochondria isolated from hamster tumors induced by adenovirus type 12 possessed circular DNA fibers. 2. The mean value of the length of the highest frequency group of the circular DNA molecules was 4.92 ±0.38 μ. 3. Catenated dimer DNA molecules and small (less than 2 μ in length) circular DNA molecules were observed.

Transformation of Japanese encephalitis antibody from IgM to IgG in the sera of the experimental infected chicks with Japanese encephalitis virus and transmission of IgM or IgG from hen to chicks were examined by the gel filtration on Sephadex G-200 column. The following results were obtained. 1. Titer of hemoagglutination inhibiting antibody rose on seven days after inoculation of mouse brain homogenate infected with Japanese encephalitis, and that increased rapidly after the second inoculation of Japanese encephalitis. The maximum peak of antibody titer attained on 35 days after the first inoculation, on 7 days after the second inoculation and it maintained for a period of 2 months then decreased. Viremia was detected till 6 hours after the first inoculation. 2. IgM antibody by gel filtration appeared on 7 days after the first inoculation, kept on rising, reached the peak on 35 days after the first inoculation, then decreased, and disappeared on 120 days. IgG antibody appeared about 2 weeks after the IgM antibody appearance, and the titer of IgG antibody became higher than that of IgM antibody on 35 days after the first inoculation, then decreased gradually, and showed 1 : 16 of titer of peak on 150 days by gel filtration. 3. We could obtain the chicks by fertilization from experimentally infected hen, having IgM and IgG of hemoagglutination inhibiting antibody of Japanese encephalitis. And the localization of antibodies in the sera of its chicks was determined by Sephadex G-200 gel filtration. And IgG antibody was detected in chick serum, though IgM antibody was not detected by this method.

To obtain some information of the biological action of Kankohso 101dinicotinate and Kankohso 301-nicotinate, observations were made on the binding mode of these substances with protein, chondroitin sulfate and nucleic acids and the following results were obtained; 1. Kankohso 10 I-dinicotinate binds reversively with bovine serum albumin or serum r-globulin, resulting in metachromasia. By binding with proteins the absorption maximum of the dye shifts toward the long wave length side and the absorbance decreased distinctly. The data show that there are more than one kind of binding sites and the binding with bovine serum albumin is weak in acidic solution and strong in alkaline solution. 2. Kankohso 10 I-dinicotinate produces strong metachromasia with sodium chondroitin sulfate and the color of the solution changes from violet blue to reddish violet. The absorption maximum at 592 mp. decreases without shifting its wave length ,and the shoulder appears at 555 mp. be. comes distinct peak. The strongest metachromatical changes occurs at the concentration of the chondroitinsulfate whose sulfonate radicals is equal to the molecules of Kankohso 10 I-dinicotinate. 3. Kankohso IOI-dinicotinate produces metachromasia with nucleic acid, where absorption spectrum is shifted toward long wave length and absorbance is decreased at a certain concentration. 4. Kankohso 301.nicotinate binds weakly with bovine serum albumin, the binding of which is reversible and the maximum binding number is 1.1 per molecule of albumin. Metachromasia cannot be produced by binding. Kankohso 30I.nicotinate does not bind with bovine serum γ-globulin. This compund does not produce metachromasia with sodium chondroitin sulfate but produces weak metachromasia with nucleic acid, indicating some affinity to nucleic acid.

The body wall of the cercaria of Schistosoma japonicum is covered with a thin integument which is connected to epithel cells located under the uscle layer. On the outer and the basal surfaces of the integument are seen thin limiting membranes. In the matrix of the integument are distributed numerous dense granules, vacuoles and spines. The rootlet of the spine is attached to the basement membrane of the integument. The circular and longitudinal muscle layers, both underlying the integument, have smooth muscle fibers composed of thick and thin myofilaments. The cercaria possesses five pairs of secretion gland cells which are divided into two groups of three anterior and two posterior pairs. Both gland cells are filled with secretion balls. The tail of cercaria is likewise covered with a thin integumen t, whose structure is identically the same as the body integument. Beneath the integument are located thin circular and longitudinal muscle layers. The circular muscle cells have smooth muscle fibers, but the longitudinal muscle cells have striated muscle fibers. These muscle cells contain many large mitochondria. On observing the cross-sections of the tail at the flame cell level the arrangement of these muscle can be divided into four muscle groups and each muscle group reveals four or five muscle cells. The excretory system is well developed and has flame cells, excretory canal and bladder.

For the purpo3e to determine exactly what stage of cell specialization the DNA level of erythroid cell nuclei begins to decline, the author observed the DNA level of erythroblasts in mitosis by microspectrophotometry and the DNA synthesis by flash labeling with H3-thymidine. The cell samples were obtained from the bone marrow of normal, blood-depleted and phenylhydrazine-treated animals, and the anemic animals received a mass red cell transfusion, all the animals being injected with colchicine 4 hours before obtaining the bone marrow sample. DNA level was measured on the smeared cells stained by Feulgen reaction and DNA synthesis by autoradiography on the smeared cells. Besides these, chromosome number was observed on the anemic rat erythroblasts at metaphase by air dry method. The observations indicated that the DNA level begins to decrease at polychromatic stage being accompanied by a decrease in TDH3-incorporation into DNA, reaching minimum level at orthochromatic cell both in DNA contents and synthesis. Chromosome numbers of erythroblasts of rat were irregular being distri buted between 42 to 20. The data have suggested that the DNA level of erythroblasts decreases only in the later stages of cell specialization, and at polychromatic stage the chromosome number may also decrease in rabbit at polychromatic stage by the cell division with an incomplete DNA replication. The high DNA level of the erythroblasts of rabbit, in severe anemia where most of the cells are denucleated at polychromatic and late basophilic stages, has been discussed from the view point of the insufficient DNA replication at polychromatic stages.

It is well known, that high-energy electrons can be used for tumor therapy. The so-called conventionel therapy with 100 through 250keV x· rays causes a great part of the x.rays to be scattered and absorbed in the sane tissue. In spite of the medicamental radiation prophylaxis additional radiation diseaes result by those compton scattered rays. By application of fast electrons and hard x.rays (so called gamma. rays) one tries to diminish those undesired side-effects and at the same time to increase the therapeutical effect of the ray treatment. As radiation source for fast electrons and hard gamma.rays one uses the Betatron, which was developed by NBRST in 1941 after preliminary operation of SLEPIAN, WALTON, WIDEROE and STEENDECK. The following statements are based on the references (1) through (6).

1. To have a rapid isolation of oligomycin-sensitive ATPase particles (OSA particles), 0.1 mg DOC per mg of protein and 72 g potassium chlo. ride per I were added to mitochondria suspended in a tris.sucrose-histidine solution, which was followed by addition of 2-fold volume of chilled water, and fractionated by a discontinuous sucrose density gradient centrifugation. As a result, it was possible to reveal the OSA particle structure, composed of the head piece, stalk and thread-like structure of a superficial portion of the base pieces, stripped off from the mitochondrial inner membrane, in a layer of density.l.lO. This fraction exhibited a remarkable activity of ATPase sensitive to oligomycin, approximately 15 ,lJ.moles Pi released per mg of protein per minute at pH 8.6 at 37° in a non-ATP regenerating assay system, and contained almost no cytochromes. 2. When the OSA particles thus isolated were heated in water bath at 65° for 2 minutes, the head pieces were detached with a concomitant loss of oligomycin-sensitivity and were purified from the supernatant by precipitation with ammonium sulfate. 3. Trypsin in low concentration slightly induced a rise in the ATPase activity of OSA particles but in higher concentration it inhibited the activity. 4. OSA particles were resistant to the treatment of urea, and it was difficult to detach the head pieces by this treatment. 5. The some fraction obtained by solubilization of thc crude OSA particles with cholate and fractionation with ammonium sulfate exhibited ATPase activity in a masked form, and the ATPase activity with oligomycin. sensitivity was restored on addition of phospholipid. 6. A discussion was made on the mode of assembly of the head pieces and associated components and biochemical properties of OSA particles.

MC-induced sarcomas produced under the skin on the back between　scapulas of C3H mice were transplanted　successively to the mice of the　same strain. Using the first and the second generation tumors, viable　tumor cells were prepared and with these tumor cells C3H mice were　inoculated. From these sensitized mice regional lymph　nodes were taken　out at certain intervals and lymph-node cells were prepared. These tumor　cells were mixed with regional lymph-node cells in the ratio of 1 : 10, and the mixed cells were transplanted subcutaneously on the back of C3H　mice, and the development and growth of tumors were observed at intervals.　As a result it was found that the inhibitory effect of these regional　lymph-node cells on the tumor growth was strong one to two weeks after　the transplantation, but beyond 3, or 4 weeks no inhibition　was observable.　In connection with the present in vivo experiments, some comments　were made on the available literature, and it has been demonstrated that　even in the cancer-bearing animal destined to die of tumors, at certain stage of cancer there is seen an inhibitory effect of the host on the tumor　growth by way of the lymphoid system and that such a response of the　host in vivo seems to be　correlated well with in vitro reaction.

Rats were depleted of skin histamine by more than 80 % by intraperitoneal injections of sinomenine with daily increasing doses for 6 days. In these rats, egg-white edema induced in the hind paws was inhibited by 68 % of control. The weight of the wall of granuloma pouch made by croton oil was also evidently smaller in the rat treated similarly with sinomenine than that of control. This suggests an important role of histamine participating in the inflammation. It has been observed that a variety of non-steroidal anti-inflammatory drugs inhibited both degranulation and histamine release induced by compound 48/80 of mast cells isolated from rat peritoneal fluid. The degranulation inhibiting actions of anti-inflammatory drugs were markedly decreased in the presence of glucose as in cases of dinitrophenol, dicumarol and warfarin which are known uncouplers of oxidative phosphorylation. Also, prevention of edema provoked by anti-rat serum is roughly correlated to a potency of degranulation inhibiting effect of anti-inflammatory agents. These observations suggest that there is a common mechanism between these two phenomena, and the prevention of mast cell degranulation by the anti-inflammatory agents is, at least, partially due to their uncoupling effects. A working hypothesis explaining the process of edema formation at the inflammatory site has. been made based on the data of the present experiment and other ob3ervations: a leakage of plasma into the tissue space from the gap between two adjacent endothelial cells which are contracted by released histamine may activate a kinin-forming system in the plasma, and kinin(s) may further aggravate a leakage. The mechanism of action of anti-inflammatory agents, which interfere with the histamine effect in inflammation, should be understood in twofold: one is prevention of histamine release from the tissue, mainly by inhibiting mast-cell degranulation, and the other is prevention of the contraction of endothial cells by their uncoupling activities.

Die Autoren berichten Uber die Forschungen mit Hilfe von Kernreaktoren auf dem Grenzgebiet der Medizin und Strahlenphysik (Reaktorphysik). Es wurden kurz ihre eigene bisherige Ergebnisse, das Ziel dieser Forschungen und einzelne Zukunftsmoglichkeiten erwahnt. Nachdem wurden in mehreren Teilen die Kernreaktoren beschrieben. 1m erstem Teil werden die allgemeinen physikalischen Grundlagen eines Kernreaktors dargestellt. Mit Hilfe der Kernkrafte und der Bindungsenergie wird gezeigt, wieso eine Kernspaltung auftreten kann und wieso aus den nicht spaltbaren Isotopen U-238 die spaltbaren Isotope Pu-239 bzw. U-233 "erbrutet" werden konnen. -Das Zustandekommen einer Kettenreaktion, die die Grundlage einer Energiegewinnung aus der Kernspaltung ist, wird erlautert, wobei sich auf die Wechselwirkungen zwischen Atomkernen und Neutronen hingewiesen wird. 1m AnschluB daran werden die Vorgange im Kernreaktor im Einzelnen naher verfolgt. Dabei werden mehrere Moglichkeiten beschriebenk die es erlauben, die Neutronenzahl, die fur die Kettenreaktion entscheidend ist, zu erhohen. Einer allgemeinen Dbersicht uber die verschiedenen Reaktortypen und ihreu Einsatz in Forschung und Industrie schlie13t sich eine Beschreibung verschiedener deutscher Reaktoren und ihrer speziellen Eigenschaften und Vorteile an. Zum SchluB wird noch besonders auf die groBen Projekte der Entwicklung von Thorium-Brutern und von schnellen Brutern eingegangen.

Exfoliative cytologic studies on normal oral cavity and on the postexodontic wound healing of infants were carried out and the following results were obtained: 1) The keratinization of cells was found marked in such regions as buccal mucosa, mucobuccal fold, gingiva and palate in that order. 2) As for changes in the distribution of cells and leucocytes, the cell distribution in the period of 15-21 postexodontic days proved to be identical with that of normal exfoliated cells. Namely, the wound healing from exfoliative cytologic aspects takes place during the period of 15-21 postexodontic days.

In order to ascertain whether black-crowned night herons (BCNH), white heron (Plumed Egrets (PE)) and domestic fowls are infected by JE virus and they serve as infection source ofJE, hemoagglutination inhibiting antibody and its 2·ME sensitive antibody in the sera of these birds were determined. Physico-chemical nature of fowl's antibody of JE produced by natural infection and their maternal antibody in the sera of chicks were examined. The results are briefly summarized as follows. 1) As to the herons captured in Tsudaka Town, two out of six adult night herons and three out of the four chicks showed positive HI reaction. On the other hand, HI reaction in the sera of two adult white herons and three chicks were negative. 2) As to the herons captured in Okayama City, twenty out of thirtytwo adult night herons and seven out of seventy white herons showed positive HI reaction in 1966 around the time when JE was prevalent in Okayama Prefecture. And six out of eleven night herons and one out of seven white herons showing positive HI reaction, responded positively to 2-ME sensitivity test. 3) The results indicate that white herons can be also infection source ofJE though less than in the case of night herons. 4) In the domestic fowls (white leghorn) kept at Takahashi District, eight out of twenty-seven fowls showed positive HI reaction. And six out of seven domestic fowls showing positive HI reaction responded positively to 2-ME sensitive reaction. 5) Transformation of JE antibody in the serum of hen from IgM to IgG was recognized. 6) Domestic chicken's sera having 1 : 640 of HI titer in the original serum and 1 : 320 of HI titer after 2-ME treatment were fractionated by gel filtration on Sephadex G-200 and the antibody activities present in the various fractions were determined. HI antibody activities occurred in both IgM and IgG classes of immunoglobulins. 7) Maternal HI antibodies reacting with JE virus were found in newly hatched domestic chickens from the eggs laid by hens with natural infection ofJE. And half life of HI antibodies in chicks was four days. 8) HI antibodies of JE in the serum of maternal immune-hens and chicken having maternal antibody were located in r-globulin fraction by starch block electrophoresis. 9) The results from 4) to 8) indicate the presence of natural infection ofJE in the domestic fowls. And domestic fowls can be infection source ofJE.

1. Both EACA and AMCHA clearly showed an anti-inflammatory effect, by intravenous, intramuscular, or oral route, against inflammatory edema produced in rats by intracutaneous injection of rabbit's anti-rat serum, carrageenin, histamine, serotonin, or bradykinin, as tested by the punch method. 2. The two compounds also showed inhibitory action against the cotton pellet granuloma when used in a larger dose. 3. There was virtually no difference between the two compounds in their anti-inflammatory activity, in spite of the fact that antiplasmin activity of AMCHA is evidently greater than that of EACA. In addition, there was no increase in fibrinolysis at the site of antiserum inflammation in rats. Therefore, it would be difficult to presume that the anti-inflammatory action of these compounds is due to their antiplasmin activity. 4. Salicylates, pyrazolidine derivatives, and non-steroidal antiinflammatory agents like flufenamic acid inhibited degranulation of isolated rat mast cells induced by compound 48/80 and also inhibited ATP-32Pi exchange reaction in rat liver mitochondria, but such actions were not observed in EACA or AMCHA. 5. Anti-inflammatory effect of EACA and AMCHA did not decrease after adrenalectomy but did become weak in hypophysectomized rats. EACA did not increase blood sugar level in normal rats so that its antiinflammatory action is not due to hyperglycemia, and the effect of hypophysectomy may not be correlated to carbohydrate metabolism. 6. Anti- inflammatory effect of EACA and AMCHA appeared more rapidly after intramuscular or oral administration than by intravenous injection but the effect was not fortified after their in vitro incubation with tissues of stomach, intestine, or liver.

A unique low density lipoprotein was obtained from the tumor transplanted with a cultured cell line of Ehrlich ascites tumor, JTC-ll cell. The tumor low density lipoprotein electrophoretically migrated as a single band, and the mobility was different from that of other organs. The chemical composition of lipid, cholesterol and phospholipids in tumor low density lipoprotein were characteristic. The flotation rate was Sf 5.9, and thus the molecular weight was estimated to be about 130 x 104. The inhibitory effect on tumor growth of the immune serum was most elevated at 25th day after the intraperitoneal administration of tumor low density lipoprotein. The main fraction effective for inhibition of tumor growth existed in γ-globulin.

Antigenicity of the peptide of ribosomal digest in Ehrlich ascites tumor was studied. The peptide was purified by DEAE Sephadex A-50 column chromatography. The peptide was electrophoretically basic, single, and 1.32 S20w sedimentation coefficient with poor content of tyrosine and phenylalanine. The maximum absorbancy was at 267 mμ. Mice and rabbits were immunized with the mixture of the purified peptide with Freund's complete adjuvant. The inhibitory effect of immune γ-globulin on the tumor growth was demonstrated in vitro cultured JTC-11 cells. A single precipitin line was observed between rabbit antiserum and tumor cell extract of Ehrlich ascites cells in ouchterlony double diffusion chamber and immunoelectrophoresis. The sedimentation coefficient of the effective fraction in immune-serum was 17 S20w. The precipitin line was observed at β2-γ region in immunoelectrophoresis.

For the purpose to reveal the relation between cell death and nuclear stainability by supravital staining with basic dyes observations have been made on the cells of bone marrow, peripheral blood and lymph node from anemic and non-anemic rabbit, rat, mouse and chicken, and thymus from young mouse. The cells were stained supravitally in blood serum, isotonic saline, calcium chloride and sucrose solutions with the dyes; brilliant cresyl blue (B. C. B), Nile blue (N. B.), neutral red (N. R.), Janus green (J.G.) and eosin (E.). The following results were obtained: 1. In the presence of blood serum all the living cell nuclei observed were not stained supravitally, except some mature erythroblasts and nucleated red cells. 2. In isotonic saline, CaCl2 and MgCl2 solutions all the erythroid cell nuclei were stained deep by B. C. B., N. B., N. R., slightly by J. G. but not by E. In stainability the younger the cell is the deeper in its nuclear staining. The nuclei of other cell strains were not stained. 3. In isotonic sucrose the nuclei of mature granulocyte were also stained by B. C. B. and N. B. but not by other dyes. The nuclei of lymphoid cells and myeloid cells appeared pale without being stained by any dyes. The nuclei of erythroblasts in sucrose solution were stained deeper with B. C. B. and N. B. than those in isotonic saline. The differences between supravital stainability of the nuclei among the cells belonging to different strain and among those of the same strain but in different maturation stage and the nuclear staining after cell death have been discussed from the possible dissociation of DNA from histone.

Antigenicity of clozapine was studied in rabbits, comparing with that of chlorpromazine as control. The results indicate that chlorpromazine produces antibody in rabbit as revealed by passive hemagglutination test, giving the titer of 1 : 2, 000 or higher in all the five cases observed, though specific precipitin lines has not been obtained and PCA test proves to be negative. Clozapine failed to produce anti.clozapine antibody giving negative passive hemagglutination test, passive cutaneous anaphylaxis and precipitin reaction, in all forms tested. Some remarks were made on the possible close relation between the antigenicity of the drug and its affinity to protein.

Urethral duplication is a rare congenital anomaly. Although a number of theories have been proposed to describe the embryology of the condition, the actual mechanism of the disorder is still unclear. We report here a case of urethral duplication in a 11-year-old boy complaining of a double stream, and review the current literature on this rare entity.

The rate of transport of blood glutamic acid into the brain and the rate of metabolic conversion of the amino acid in the brain were derived by the use of the brain perfution method in vivo and in situ with [D.HC] ·Lglutamic acid. The net uptake of glutamic acid by the brain was observed. Most of the radioactivity released from the brain into the cerebral venous blood was found to consist of glutamine. Small but significant amounts of output as radioactive GSH and CO2 were also found. Glutamic acid transport and its rate of metabolism were lowered in the glucose. free condition. The size of the compartment of the small glutamic acid pool, which was related closely to the blood glutamic acid, and that of the large glutamic acid pool, which was related closely to the blood glucose, were calculated and compared with each other.

1. Marsupialization was performed on 23 cases of cystic lesions of the jaw at the Department of Oral Surgery, Okayama University Hospital in the recent five years. 2. These patients were divided into 4 groups according to their age; namely, group A of those under IS years old, group B of those between 15 and 30 years old, group C of those between 31 and 60 years old, and group D of those over 61 years old, and the results of postoperative findings were compared with those of preoperative ones. 3. In group A of the four groups the most favorable results were obtained after marsupialization and reduction of the tumor was remarkable, even in a case of ameloblastoma, as compared with groups B, C and D. 4. Cure of lesions took somewhat a longer period of time in group B than in group A, but all the cystic lesions were reduced favorably after operation. 5. The reduction in cysts in groups C and D was markedly slower when compared with that in groups A and B, but the marsupialization surgery seems to be desirable in some cases.