1. Electron microscopic observations were made on DNA fibers in the mitochondria of tumor cells induced by various kinds of DNA viruses, RNA viruses, and chemical carcinogens, and also in those of normal and regenerating rat livers and cultured liver cells. Intramitochondrial DNA fibers were observed most frequently in the sectioned specimens of adenovirus type 12-induced or SV 40-induced hamster tumor cells, but were hardly observed in normal and regenerating liver cells and other tumor cells.
2. The DNA fibers disappeared by treatment with DNase, but not with RNase.
3. The DNA fibers in adenovirus-induced tumor cells were easily isolated by osmotic shock and observed as circular DNA molecules by rotary shadowing with the electron microscope, while DNA molecules were hardly isolated from rat liver mitochondria by the same treatment.
4. On the ultracentrifugal fractionation of sonically disrupted adenovirus-induced tumor mitochondria, DNA were found to be contained mostly in the supernatant fraction by chemical analysis and by electron microscopic observation.
5. DNA fibers observed in the mitochondrial matrix in the sectioned specimens were proved to be identical with the isolated circular DNA molecules.
6. On the ultracentrifugal fractionation of sonically disrupted rat liver mitochondria, DNA were found to be contained mostly in the membrane fractions by chemical analysis. However, DNA molecules were difficult to observe with the electron microscope in the sectioned specimens of the rat liver mitochondrial membrane fractions. Nevertheless, they were isolated by phenol extraction from the membrane fractions, and were observed as circular DNA molecules with the electron microscope.
7. Appearance of the DNA fibers in the mitochondrial matrix seems to be associated with the division cycle of mitochondria, and in the mitochondria where DNA fibers are not observed in the sectioned specimens, the DNA molecules are supposed to be hidden by firmly attaching to the inner membrane, and to be isolated by chemical extraction.

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The diagnosis of disseminated parapancreatic calculus is relatively easy from its specific roentgenogram, but in those cases with little roentgenographic shadow or with a solitary shadow it becomes quite difficult. Hence in the latter cases it is necessary to distinguish it from bilestone, kidney stone, peritoneal calcified lymph node, splenic venous thrombus, or duodenal diverticulosis. Therefore, for the diagnosis of the parapancreatic calculus it is important to take a single roentgenogram of the entire abdomen as well as lateral views of the abdomen, the combination of which would afford us to identify the parapancreatic calculus from the density of its shadow, its site and its size. We have recently encountered three patients with pancreatic calculus and present our observations of the cases as well as some remarks on them from the aspects of roentgenology.

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A histochemical and electron microscopic study of the cytological features of the fiber types in normal human intercostal muscle is presented. From observations on succinic dehydrogenase, Sudan staining, and by electron microscopy, the human striated muscle fibers can be divided into three biber types; red, white and intermediate fibers. On the basis of mitochondrial content and distribution, each fiber type is characterized as follows.
1) The size and the composition of mitochondrial chains among the myofibrillar spaces: large and long in the red, absent in the white and medium in the intermediate fiber.
2) The size and shape of bracelet-like mitochondria which encompass the myofibrils at each I-band tend to be plump and multiform in the red, but slender and elongated in the white and the intermediate fibers.
3) Deposits of mitochondria beneath the sarcolemma, which are numerous in the red, scarce in the white and moderate in the intermediate fiber.
Attention has been further paid to the sarcoplasmic reticulum, lipid droplets and glycogen particles.

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An electron microscopic study was made on the fine structural differences in the motor endplates among the red, white and intermediate muscle fibers of the human intercostal muscles and the following results were obtained:
1) In the motor endplate of the red fiber, the junctional folds appear on the whole as simple comparatively unbranched holds, while in that of the white one they are branched and anastomosed in a complicated manner. In the motor endplate of the intermediate fiber, the character of the folds is intermediate between those of the red and white fibers.
2) The sarcoplasmic area where the axonbranchlet is situated is a apparently narrower in the red fiber than in the white or intermediate fibers.
3) The terminal branchlet in the red fiber is oval in shape but in the white one it has a concavity oriented towards the underlying muscle fiber. In the intermediate fiber the branchlet is intermediate between those of the red and white fibers.
Attention has been further paid to the myofiber structure particularly adjacent to the motor endplate.

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Sal. typhi, Sal. paratyphi A and Sal. paratyphi B which are the representative strains of group D, A snd B of Salmonella group respectively, have special characteristics of inducing typhoid fever in man. Their complex antigenic structures and biochemical characters are already well known.
The authors studied the difference, if any, in the lipid components among these strains, while comparing with the lipid components of other Enterobacteria.
1. The major components of phospholipid were found to be identical in all the three strains. Moreover, these were almost the same in E. coli and Sal. typhimurium, with an exception of a minor unknown component in E. coli.
2. Considering the composition of individual components, three strains contained a considerable amount of cardiolipin (10-13% of total phospholipid). On the other hand, they contained phosphatidyl glycerol in several to 10%.
3. Among the three strains, Sal. paratyphi B tended to show the quautities of cardiolipin and phosphatidyl glycerol very similar to those in E. coli B.
4. The fatty acid composition of total lipids in these three strains did not differ so appreciably from that of E. coli. Main components of the fatty acid were palmitic, palmitoleic and oleic acids.

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The carbohydrate metabolism in the carcinogenesis of DAB fed rat hepatoma was investigated by studying the G-6-Pase and Glucokinase activities to analyse the marked increase of glycolysis and decrease of respiration in the cancer tissue. And same time, these enzymes activities were measured in embrionic, new born, and regeneratig rat liver, which showed rapid growth and cell division, to study whether there was biochemical specificity in hepatoma.
The liver homogenate G-6-Pase activity decreased progressively along with DAB feeding and was almost absent in the hepatoma tissue, while the glucokinase activity increased gradually and about 4 times of normal activity in cancer tissue.
On the other hand, there was no change of glucokinase activity in the embrionic, new born, regenerating, or normal rat liver. And the G-6-Pase activity was almost absent in the embrionic rat liver and showed rapid increase after birth to maximum in 12th day born rat liver.
From these results, both enzymes have important significance in the elevated glycolysis in cancer, although only G-6-Pase seems rate limiting factor in the physiologically elevated glycolysis state of the liver. So difinite difference was recognized between hepatoma and these tissues biochemically.
The serum G-6-Pase activities in the carcinogenesis of DAB fed rat showed no significant change from normal rat.
For clinical application, the serum G-6-Pase activities of various patients were also measured. Patients with hepatic disturbance especially acute hepatitis or active hepatitis demonstrated significant elevation of the G-6-Pase activity while no other patient did not. So the measurement of serum G-6-Pase activity is very useful in clinical aspect.

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Cortical evoked responses after single or repetitive stimulations of the nucleus ventralis lateralis (VL), n. centrum medianum (CM) and n. ventralis anterior (VA) were observed in 61 cases of parkinsonism, 7 cases of intractable pain and a case of dystonia musculorum deformans from February, 1968 to December, 1969. Experimental studies of the thalamic stimulations with 15 cats were performed as well and the results were compared with that of the human. Evoked responses were analysed with averaging computer technique. Results were as followings.
1) Cortical evoked potentials after single stimulation of the VL nucleus of the human thalamus.
a) Evoked potentials appeared on the ipsilateral centro-frontal regions with weaker stimulation in EEG records. However, the evoked potentials showed clear bilateral distribution except the temporal region, predominantly on both centro-frontal regions, with stronger stimulation electroencephalogaphically.
b) The evoked potentials of the central site showed clear three negative waves within 200 msec after the stimulation, that is, the first negative wave (I-N) with the peak time of about 10 msec, the second (positive-) negative wave (II-PN) with the peak time of about 50 msec and the third negative wave (III-N) with that of 80-100 msec with averaging computer records. The peak time of the I-N wave and the negative phase of the II-PN wave were nearly constant, but that of the III-N wave considerably varied according to the back-ground activities of EEG. The evoked potentials showed no significant discrepancy between right and left patterns, and moreover, they maintained a similar pattern when the stimulating voltages were changed.
c) Bilateral evoked potentials were clearly seen with averaging computer technique even when evoked potentials were small and hardly identified on the contralateral scalp with EEG records. When the stimulating voltages were gradually decreased, the ipsilateral and contralateral evoked potentials disappeared approximately at the same time.
d) The latency of the I-N wave was measured to be 5-6 msec with averaging computer records, and the amplitude of the I-N wave increased proportionately with the increase of stimulating voltages.
e) The negative phage of the II-PN wave was the most dominant component of the three negative waves and the amplitude was highest when the stimulating point was thought to be in the center of the VL nucleus, and the II-PN wave disappeared when the point was outside of the thalamus.
f) Simultaneous recordings of the evoked potentials at the bilateral central surfaces as well as several intracortical deep points were performed. I-N wave reversed its polarity in the depth of the cortex, namely a clear positive wave, whose peak time was corresponded to that of the I-N wave on the cortical surface, was seen in the depth of the cortex. Positive and negative phase of the II-PN wave on the cortical surface reversed their polarity after passing through so called “The zone of isopotentiality” as well.
2) Augmenting and recruiting responses after low frequency stimulations of the human thalamic nuclei.
a) Low frequency stimulation (6-13c/sec) of the VL nucleus of the human thalamus usually evoked clear bilateral augmenting responses, prodominantly on both centro-frontal regions.
b) There were particular two parkinsonian patients who showed no evoked responses at single stimulation of the VL nucleus with 10 volts and more but showed clear recruiting-like responses at low frequency stimulation of the VL nucleus.
c) Low frequency stimulation of the CM nucleus usually evoked bilateral, wide distributed menophasic negative responses, predominantly on both parietal regions, that is, the CM nucleus showed typical recruiting responses. Low frequency stimulation of the VA nucleus evoked the recruiting responses. with predominant appearance on both centro-frontal regions.

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The synthesis of fatty acids and their esterification by blood from 12 cases of liver cirrhosis was studied in terms of incorporation of ¹⁴C from 1-¹⁴C acetate into individual fatty acids in comparison with those found in the groups of 5 chemical diabetics, 5 overt diabetics, 6 cases of acute hepatitis, and 11 normal subjects. Of the patients of liver cirrhosis 6 cases were abnormal in glucose tolerance test (GTT) in spite of uncertain complication of primary diabetes and the rest was clinically considered to have been complicated with primary diabetes.
The results were as follows.
1. It was proved in such major lipid classes as triglyceride (TG), Phospholipid (PL), and free fatty acids (FFA) that the depression of absolute distribution of the labelled fatty acids synthesized was found in every group of diseases as compared to normal controls, however in both TG and FFA fractions the most striking depression was found in the groups of liver cirrhosis with abnormal GTT, overt diabetics, and acute hepatitis. While the remarkable decrease in PL fraction was found in every group of liver diseases particularly in the groups of liver cirrhosis.
2. It was found in the diabetic groups that the percentage of synthesized fatty acids distributed to TG was lower and that distributed to PL was higher than normal group, on the contrary the percentage distributed to TG was higher and that distributed to PL was lower than normal group in both groups of liver cirrhosis.
3. Both groups of liver cirrhosis showed the striking increase of percentage distribution in 18: 1 as compared to not only normal but diabetic groups and that this change was found in every fraction of TG, PL, and FFA. However the increase of the percentage distribution in 18: 1 was most strikingly reflected to TG fraction, because the esterification of 18: 1 was most specific for TG fraction and on the other hand the percentage of this fatty acid in the diabetic groups showed rather lower values as found in the normal group. In patients of liver cirrhosis the striking increase of the percentage distribution in 18: 1 was weakened to some extent by complcating the primary diabetes in all lipid classes examined, nevertheless the pattern was rather analogous to that of liver cirrhosis than diabetics.
The most part of 14: 0 and 16: 0 remained unesterified and the percentage distribution of these fatty acids in FFA or PL fraction was lower in the groups of liver cirrhosis and diabetics than that of normal controls or acute hepatitis. The esterification of fatty acids with retention time corresponding to 20: 0 or longer was most specific for both TG and PL fractions and in these fractions the percentage distribution of these fatty acids increased in the diabetic groups and decreased on the contrary in the groups of liver diseases as compared to normal group. No particular differences were found among each group in the percentage of 16: 1 and 18: 0 in every lipid classes.
4. In acute hepatitis the fatty acid synthesis by blood showed some quantitative decrease, little qualitative changes in the percentage distribution of individual fatty acids were found in every lipid class as compared to normal group.

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The fatty acid compositions of whole blood from 12 cases of liver cirrhosis were studied from the view of total and fractionated major lipid classes in comparison with those of 5 overt diabetics, 6 cases of acute hepatitis, and 11 normal subjects. The patients of liver cirrhosis were classified into two groups; the one group of 6 cases was clinically considered to have been complicated with primary diabetes and the rest were uncertain whether the diabetes had complicated or not in spite of having abnormal glucose tolerance.
The results were as follows.
1. It was found in the group of liver cirrhosis with abnormal glucose tolerance that the increase of percentage composition in 16: 1 and decrease in 18: 2 were most specific changes, besides not only the tendency of increase in 16: 0 and 18: 1 but that of decrease in 20: 4 was also found in the total lipids. The similar changes were observed in the groups of liver cirrhosis with primary diabetes and acute hepatites. On the other hand, the main changes found in the diabetic group were increase of percentage compositions in 16: 0 and 18: 1.
2. The changes of fatty acid composition in cholesterol ester were found to have most directly reflected to those found in the total lipids in the group of liver cirrhosis, while the changes in the triglycerides reflected directly to those of total lipids.
3. Such tendencies were found in the groups of liver cirrhosis with abnormal glucose tolerance as the increase in 16: 1 and 18: 1 as well as the decrease in 18: 2 and 20: 4 in the percentage composition of cholesterol ester, while the increase in 16: 0 and the decrease in 18: 2 in triglycerides, the decrease in 18: 0 and the increase in 18: 2 in phospholipid, and the decrease in 20: 4 in free fatty acids. The almost same tendencies were also observed in the groups of liver cirrhosis with primary diabetes or acute hepatitis.
The main change found in the cholesterol ester of diabetic group was increase of percentage composition in 18: 1 and the almost same changes as in the groups of liver diseases were revealed in other lipid classes.

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Antigenicity of ribosomal digest of Ehrlich ascites tumor was studied in vivo. Antitumor activity was observed by intraperitoneal transplantation of Ehrlich ascites cells, comparing with body weight between control group and immunized ones, which were treated with tumor ribosomal digest.
The body weight increasing curve in case of 100500×10⁴ Ehrlich ascites cells inoculum was no visible differences between control group and immunized ones.
The antitumor effect was visible in case of 50×10⁴ tumor cells inoculum, the LD50 of immunized group was 16 days and that of ontrol 14 days. The effect was more clearly observed in case of 10×10⁴ tumor cells inoculum. The LD50 of immunized group was 17 days and that of control 14 days.
No visible effect were in mice transplanted with Ehrlich ascites cells on the back between control and immunized group.
Neutralization experiment with rabbit anti-serum prepared by Ehrlich ascites tumor ribosomal digest was observed by multiple dilution method. The effect was most strong at 0.2ml of anti-serum. for 50×10⁴ tumor cells.

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Immuno-electrophoresis and precipitin Reaction were performed by using rabbit antiserum and ribosomal digest of Ehrlich ascites tumor. A single precipitin line was observed by double diffusion test (Ouchterlony Method) between tumor ribosomal digest and ribbit antisesum.
The precipitin line was immunoelectrophoretically moved from β₂ to γ₁ position.
The titer of rabbit antiserum was 16x and precipitin line was most clearly appeared at 1-0.5 mg/ml protein concentration.
Equivalent region of antiserum protein concentration was 0.7 mg/ml in quantitative precipition reaction. γ globulin of rabbit antiserum was prepared by DEAE-Sephadex A 50 colum, and the sedimentation coefficient was 17.05 S_20w in Ultracentrifugal analysis. Electrophoretically the γ globulin was moved from β₂ to γ₁ position. Therefore, this γ globulin is to be Ig M.
A single precipitin line was observed in Ouchterlony method between γ globulin and ribosomal digest. This γ globulin would be immunologically active form.

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