Okayama University Medical School

1. The mixed lymphocyte culture test where live lymphocytes of female mice are cultured with supersonicated cell homogenate of isogeneic male mice does not reflect the difference in H-Y histocompatibility antigen alone. 2. When non-H-2 antigens are cumulated in various combinations, the rate of blastformation becomes greater than the combination of C3H ♀ themselves. 3. When non-H-2 antigens and H-Y antigens are added in various combinations, the rate of blastformation becomes significantly higher than in the combination of C3H ♀ themselves.

For the purpose of forecasting the prevalence ofJapanese encephalitis in Japan, we tried to find out the correlation of factors between median and mode dates of epidemic time curve of prevalence on one hand, and average atmospheric temperatures of prefectures in June and July (T6,7 in short) (X¹), the time when HI reaction of swine became positive to the degree of 50 per cent (D. pos. swine in short) (X²), the latitude (x³) and longitude (x4) in respective prefectures (in 1965 and 1967). On the other we also estimated the median and mode dates of this epidemic curve of the prevalence in 1968 and 1969, from the regression equation of one variable and multiple regression equation from the above factors using an electronic computer. The usefulness of adding factors concerned with mosquitoes to the above four factors is proven by the accuracy of estimation. And the following results were obtained. 1) Phenomenally speaking, the prevalence of Japanese encephalitis follows the principle of "advancing of prevalence towards the north and east" and essentially speaking, it depends upon high atmospheric temperature and the outbreak of many hazardous mosquitoes by the high atmospheric temperature. 2) To estimate median date (y) and mode rate (z) of the epidemic time curve of the prevalence, we can use the next equations; The regression equations to estimate y and z from T 6,7(X) are as follows. y = - 3. 75X¹ + 144.47 σ = 12.4.·. [1] z = - 3. 80X¹ + 157 .26 σ = 14.9.. · [1]' The regression equation from D. pos. swine (X²) are as follows. y = 0. 68X² + 31. 82 σ = 9.2· .. [2] z=0. 76X² +40. 71 σ= 12.0 .. · [2]' The multiple regression equation from T6 ,7 and D. pos. swme are as follows. y = -1. 07X¹ +0 .62x² +59. 37 σ= 9.7 ... [3] z= -0. 79x¹ +0. 71x² +61.02 σ= 12.0· .. [3]' The multiple regression equations from T 6•7, D. pos. swine, latitude and longitude are as follows Y= -1.01x¹ +0.58x² -0.26x³+0 .37x4 + 18.50 σ= 9.8･･･ [4] z = -0. 32x¹ +0. 52x² +2 .05x³ +0 .54x4 -87. 81 σ= 11.8 [4]' 3) We Obtained the estimated value of median date in 17 prefectures in Kyushu, Chugoku, Shikoku, Kinki and Kanto provinces in 1968 and in 13 prefectures in 1969 from [l] or [2] or [3] or [4] equation. Nine prefectures out of 17 by [l], 12 prefectures by [2], 13 by [3J and [4] in 1968. [4] could be estimated with about 10 days error or less. And in 1969, 9 out of 13 by [3] and 7 out of 13 by [4] could be accurately esti· mated. The estimation by the multiple regression equation using many factors is most useful for the calculation. 4) The time when the number of patients increases at maximum can be pointed out by the lower limit of prediction region obtained from data in each prefecture. And the lower limit was the estimated median value minus about 20 days by [1] and about 16 days by [2] or [3] or [4] under the next condition; α = 0. 1, N= 75. 5) The mode dates in 17 prefectures out of 19 were estimated by [1]', [2]', [3]' and [4]'. 12 prefectures out of 17 by [1]', 7 by [2]', 10 by [3]' and 13 by [4]' could be estimated with about 12 days error or less in 1968 and 9 out of 13 was correctly estimated by [3]' and [4]' in 1969. The estimation by the regression line of one factor was s~mewhat different from each other, but when multiple regression line of four factors was used the estimation became more correct. Judging from these results, it is adequate to use the multiple regression equation of [4] and [4]' when we want to forecast the median date or mode date ofJapanese encephalitis time cure. 6) In the case of adding two factors concerned with mosquitoes to T6,7 (X¹), D. pos. swine (x²), latitude (x³), longitude (x4), multiple regression equations become as follows. y= -1.46x¹+0.14X²+0.068x5+89.03 σ= 6.9.. ·[5] z= -3. 29x¹+0 .13x²-0. 010x5+ 143.63 σ= 18.6··· [5]' y=-4.20x¹+0.35x²+0.29x6 + 53.70 σ= 4.2 .. ·[6] z=-2.56x¹-0.0lx²-0.02x6 +128.96 σ=11.4 [6]' y= 4.76x¹+0.41x²+0.13x5+0.22x6-72.78 σ= 4.5 [7] z = - 2. l0x¹ + 0. 05x²+ 0. 11 x5 - 0. 08x6+ 113.4 σ= 10. 7.. · [7]' where x5 is the time when the number of mosquitoes (C. T. collected by light trap reached the maximum and X6 is the time when hazardous mosq uitoes were dected. In the case of median date, 5 prefectures out of 6 prefectures by [5], 2 out of 6 by [6] and 2 out of 5 by [7], and in the case of mode date, 5 out of 6 by [5]', 4 out of 5 by [6]' and 4 out of 5 by [7]' could be accurately estimated in 1969.

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.

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.

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.

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.

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.

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.

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).

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.

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.

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.

For the purpose to obtain the information of the mechanism of protein uptake by the tumor cells, some cytochemical and electron microscopic observations were carried out on Ehrlich ascites tumor cells incubated with horseradish peroxidase (basic hemoprotein, molecular weight approximately 40,000) in vitro. In the earlier periods of the incubation peroxidase was found to be adsorbed on some area of surface of the tumor cells forming a thin protein layer, where an active pseudopodia formation was observed. With the lapse of time, the protein was taken in the deep cytoplasm by the infoldings of the cell membrane and accumulated in the cytoplasmic vesicles having limiting membrane. Concerning the accumulation of the protein into the vesicles, small tubular structures in the cytoplasm connecting the cell surface and the vesicles, were considered to participate in the intracellular transportation of peroxidase taken up. In cold environment (2°C), the formation of pseudopodia and deep inward infoldings of the cell membrane was inhibited and simultaneously the uptake of peroxidase stopped. Iodoacetate and sodium fluoride also effected to suppress the pseudopodia and infoldings formation moderetely, as well as uptake of peroxidase, though they did not stop completely. These facts have indicated that horseradish peroxdase is taken up by Ehrlich ascites tuimor cells through pinocytosis which involves energy-requiring process dependent upon glycolytic metabolism of the tumor cells.

With the purpose to clarified the mode of localization and mechanisms of activation of ATPase in the mitochondrial membrane, analyses were made on the properties of mitochondrial ATPase from the structural and functional aspects. The activation of ATPase by DNP and Mg++ and the oligomycin sensitivity were investigated in a series of inner membrane fragment samples obtained by ultrasonic irradiation and those samples obtained in the processes of isolation and purification of ATPase from rat liver mitochondria and beef heart mitochondria in parallel with electron microscope observations. As a result it has been found that the membrane fragments obtained from rat liver and beef heart mitochondria by ultrasonication exhibited high respiratory activity and unmasked ATPase activity which was charac· terized by remarkable stimulation by Mg++ and inhibition by oligomycin and azide. Therefore, mitochondrial ATPase seems to be bound fairly closely to the inner mitochondrial membrane. In the membrane fragments prepared by ultrasonication of intact mitochondria, ATPase activity was stimulated by DNP, but in the supernatant fractions was not. On the other hand, the supernatant fraction obtained from BHM and inner membrane fragments by severe sonication exhibits a marked ATPase activity and the activity incresed in each step of the purification on the treatments with acid, protamine and heat. Especially in the case of membrane fragments the protamine treatment can be omitted. Electron microscope observation of the fractions in each step of the purification proved the head pieces to be ATPase. The ATPase activity of solubilized head pieces is insensitive to oligo. mycin and coincides with the soluble ATPase of PULLMAN etat. (8) in the points of its cold labile property and optimum pH, but it shown no accele. ration of ATPase activity by DNP.

The role of -SH groups in mitochondrial energy transfer reaction was studied by observing the reduction of a disulphide, 5, 5'-dithiobis (2-nitrobenzoic acid), DTNB, a specific analytical agent for the estimation of -SH groups in biological materials, by addition of it to the isolated rat liver mitochondria in various respiratory states, as defined by CHANCE and WILLIAMS. 1. In the various respiratory states, states 1 to 5, the reduction of DTNB proceeds most rapidly at state 5, and most slowly at state 3. DTNB reduction at state 5 is suppressed by the partial oxidation of respiratory carriers with oxygen (state 4) and the addition of respiratory substrate does not affect the DTNB reduction. 2. The retardation in the reduction rate at state 3 is relieved partially by a respiratory inhibitor, KCN, and is intensified markedly by oligomycin, an inhibitor of oxidative phosphorylation. An uncoupler for oxidative phosphorylation, DNP, does not affect the reduction rate at state 3. At state 4 the reduction is stimulated by DNP and KCN, but is unaffected by oligomycin. The results suggest that the alteration in the functions of the energy transfer reaction in mitochondria is accompanied by changes in the occurrence and the functioning of -SH groups which can be detected by the reactivity with DTNB. The data suggest also that there are at least two kinds of -SH groups reacting with DTNB: the one is the -SH group which reacts DTNB actively when the respiratory carriers are kept reduced, and the other is the one which reacts actively when the respiratory carriers are kept oxidized, participating in the phosphorylating system and its reactivity with DTNB diminishes in the actively phosphorylating states (states 2 and 3).

Eine Salieyl-Chinin-Lithium-Kombination wurde im Siemens Unterriehtsreaktor SUR 100 BE des Instituts fur Kernteehnik der Teehnisehen UniversiUit Berlin mit Neutronen aktiviert. AnsehlieBend erfolgte die Messung der spektralen Verteilung der Gamma-Zahlrate zu zwei versehiedenen Zeitpunkten naeh der Aktivierung. Es wird festgestellt, daB diese Salieyl-Chinin-Lithium-Kombination nur sehwaeh im SUR-Reaktor aktivierbar ist, vergliehen mit einer Goldsonde etwa 200 mal sehwaeher. Trotzdem konnten mit Hilfe eines 400-Kanal-Analysators die spektralen Verteilungen der Gamma-Energien dieses Praparates noch sehr genau bestimmt werden. Ein ausgepragter Gamma-Peak tritt auf bei der Energie 0.53 MeV. Er klingt ab mit einer Halbwertszeit von 1.6 h und ist vermut· lich auf ein Element der Tablettierhilfsstoffe zurUckzufUhren. Vom strahlenphysikalischen Gesichtspunkt ist das Praparat fUr die Human. medizin gut geeignet.

1) It has been found that the peripheral blood of hybrid adult dogs contains about 29.4 % of lymphocytes in average. When such a blood is passed through the glass wool column the leucocytes containing 68-90 % (81.6% in average) lymphocytes are obtained. 2) In the single culture of such lymphocytes alone and mixed culture both live lymphocytes and sonicated lymphocytes in the presence of 1% (vIv) PHA, the peak of the blastformation of lymphocytes is observed at culture hour 72. 3) In the abscence of PHA both single culture and mixed culture of lymphocytes show hardly any blastformation. 4) In the mixed culture of live lymphocytes with homogenate of sonicated lymphocytes with addition of 1% (vIv) PHA the rate of blast. formation observable at culture hour 72 and the rejection of kidney trans· plant 7 days after its transplantation of hybrid adult dogs show a direct correlation, demonstrating that the mixed lymphocyte reaction reflects accurately the difference in donor histocompatibility antigens against the recipient.

For the purpose to reveal the changes in the metabolism of erythroblast in varied specialization stages the author observed the Feulgen DNA level of rabbit erythroblasts by microspectrophotometry. Observations were made on normal and anemic animals, and those receiving a mass red cell transfusion at the recovery stage of anemia where the early denucleation is stimulated. Observations have revealed that in normal erythropoiesis the DNA contents are kept at n to 2 n level from the proerythroblast to late basophilic stage, but in later stages, polychromatic and orthochromatic, DNA level per cell decreases gradually with advance of the cell specialization reaching the minimum level, nearly haploid level, at orthochromatic stage where most cells are believed to be denucleated. In blood depleted animals nearly the same pattern of DNA level was observed in connection with erythroid specialization as that in normal animal, except a relatively high DNA level in the later specialization. In the cases of the hemolytic anemia a similar tendency has been observed but the minimum level of DNA remains at a higher level, hypo-diploid level, in poly- and orthochromatic stages. Twenty-four hours after the mass red cell transfusion by which severe anemia has been recovered to the original level within one hour, the pattern of the DNA level of the erythroblast returns to the normal one showing a very low DNA level at the polyand the orthochromatic stages. The data indicate that the DNA synthesis of erythroblast kept at n to 2 n levels until the late basophilic stage begins to decline at polychromatic stage and reaches nearly haploid level at orthochromatic stage, but in active hemopoiesis the DNA synthesis is stimulated and the DNA contents are kept at a high level even in the late specialization stages, showing no relation between the denucleation and the low DNA level.

Localization of IgM and IgG sypylitic antibodies in the sera of patients and the experimental syphylitic rabbits was examined by the gel filtration on Sephadex G.200 column. I) In the case of late syphylitic patients; OGATA test-reactive antibodies were contained in IgM and IgG fractions. On the other hand, RPCF test-reactive antibody was contained only in IgG fraction. This discrepancy may be due to the difference in antigens; Cardiolipin.resicin and T. P. Reiter protein. 2) In the case of the experimental syphylitic rabbits; The results were as follows. a) Variation in the level of the titer. The peaks of the titer were seen 3-4 weeks after inoculation of T. P. Nichols by OGATA test, VDRL test and RPCF test, thereafter titers decreased. On the other hand, the titer kept on rising up to 2 months and maintained high level during the periods of 3, 4 and 5 months by TPHA test. b)Transformation of antibody from IgM to IgG. Transformation of antibody from IgM to IgG was seen 3-4 weeks after inoculation by all four tests; OGATA test, VDRL test, RPCF test and TPHA test, and such a transformation was completed 3 months after inoculation.