A therapeutic and prophylactic effect of α-mercaptopropionyl glycine for toxicity of mercury and its compounds was tested in comparison with the effect of D-penicillamine. The nine subjects working in an industrial factory manufacturing insecticides made from mercury were selected for the investigation. The daily dosis of α-mercaptopropionyl glycine was 450 mg and it was 1000 mg of D-penicillamine. The results obtaind were as follows:
1. The average daily excretion of mercury in urine was increased 1.4 to 1.6 times (average, 1.5 times) of the amount estimated before the administration when the subjects were in exposure of mercury. The increase was more than that obtained with the use of D-penicillamine (average, 1.2 times).
2. The average daily excretion of mercury in urine was increased 2 to 3 times of the amount estimated before the administration when the urinary excretion of mercury was determined after the exposure. The increase was lasted as long as 12 days after the discontinuation of the exposure. α-Mercaptopropionyl glycine was proved to be more effective than D-penicillamine when it was tested serially in a same subject.
3. From the results obtained here, it was concluded that the administration of α-mercaptopropionyl glycine at the daily dosis of 450 mg was proved to be effective for the prophylaxis of mercury toxicity. It was especially true when the sujects were working in the exposure of mercury, therefore, it was recommended to use α-mercaptopropionyl glycine as long as the increase of urinary excretion of mercury was no more observed.
4. During the administration, there observed no appreciable side effects, and so the prophylactic use of α-mercaptopropionyl glycine was proved to be not hazardous.

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Δ⁴-Cholestenone-4-C¹⁴ metabolism in male Wister rat was studied. Δ⁴-Cholestenone was not esterified, was metabolized to cholestanone, and then cholestanol. This passway was investigated on thin layer chromatography and radioautogram. In tumor-bearing rat, the metabolism observed about 1.8 times than that of normal, and cholestanone was metabolized 1.5 times greater than in normal.

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Δ⁴-Cholestenone-4-C¹⁴ metabolisms were studied with the nuclear, mitochondrial, microsomal and supernatant fraction. Enzymic activity of cholestenone to cholestanone was higher in nuclear and mitochondrial fraction, and the activity was higher about 2-4 times in tumor-bearing rat. This suggests the enzyme block from cholestenone to cholestanone. In aceton powder, these enzymes were inactivated, and the enzymes were exsisted in the fraction precipitated with the concentration of (NH₄)₂SO₄ at 30 g/dl.

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1) A case reported of Wilson's disease confirmed by autopsy was presented.
2) The patient was a 17-year old girl. The disease developed with dysarthria having tremors like Parkinson's disease on the left hand and after passing subacute stage, the symptoms of extrapyramidal tract aggravated. Later in the terminal stage there occurred two attacks of clouding of consciousness and finally she died in the state of coma with high fever. However, Kayser-Fleischer ring was not detected.
3) The autopsy examination disclosed coarse nodular liver cirrhosis which is typical of Wilson's disease. Macroscopically, there was observed no marked change in the lentiform nucleus, but microscopically, there were such findings as Wilson's elements and pseudosclerotic elements that corresponded to histopathological changes of Wilson's disease.
4) Further discussion was made on the results of histochemical studies on copper, iron, polysaccharides and sugars.

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Clinical, cerebro-pathological, histochemical and biochemical studies were carried out on a case what appeared to be the special type (Inose) of hepato-cerebral disease, and the following results were obtained.
1) Clinically, there could be recognized no Kayser-Fleischer ring, and the liver disturbances were observed from early stage, edema and ascites being especially marked. In addition, characteristic tremors of the fingers, i.e. Wackel tremors, were observed. It was characteristic with this case that there occurred repeated clouding of consciousness for the four months prior to death.
2) There were observed abnormalities in the copper metabolism. Namely, despite the presence of copper deposition in the cytoplasm of liver cells, no such deposition could be detected in the brain tissue. Furthermore, tbe amount of copper excreted into urine proved to be as much as twice that in normal person.
3) From the pathology of the brain, Alzheimer type II glia could be seen everywhere in the brain, and globus pallidus as well as the boundary between the cerebral cortex and medullar had fallen into a marked spongy state. In addition, carmine-positive substance was found in the nuclei of liver cells and in Alzheimer type II glia nuclei. These finding correspond to those of the special type of hepato-cerebral disease as reported by Inose. However, there were also observed Opalski cells in putamen and globus pallidus, the finding said to be specific to Wilson's disease.
4) Pathologically, the liver was in the state of coarse, nodular and annular cirrhosis, and the fat degeneration of liver parenchyma was marked. By the iron staining both the liver and brain gave negative results,
5) Some comments were made on hepatic coma, Wilson's disease and the special type of hepato-cerebral disease, while referring to the available literature, and also discrepancy in regard to the findings in these diseases was pointed out.
6) Admitting that the abnormality in the copper metabolism, the pathological findings of the liver and the appearance of Opalski cells would raise some question of doubt, after due consideration on these points it seems more rational to classify this case as the special type (Inose) of hepato-cerebral disease.

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Male Donryu rats were fed with 3-methyl aminoazobenzene (DAB). Phosphatidic acid m tabolism in the liver at 20, 40, 60, 80 and 100th day of DAB-feeding rats was studied and compared with the normal ones. The phosphatidic acid metabolism, especially, the pass way of diglyceride synthesis in DAB-feeding rat liver with the phosphatidic acid phosphatase studied in vitro. The fed with 3-methyl-DAB was induced hepatoma on the 40th day and the grades of hepatomas were gradually accelerated. At 80th day, livers were almost occupied with hepatomas. The enzymatic activity of phosphatidic acid phosphatase was most elevated at 80th day of DAB-feeding rat liver.

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Male Donryu rats were fed with 3 methyl diaminoazobenzen (DAB). The liver was homogenized and incubated with ³²P-phosphatidic acid at 20, 40, 60, 80 and 100th day. The distributions of ³²P freed from phosphatidic acid by phosphatidic acid phosphatase were studied on lysolecithin, sphingomyelin, phosphatidylcholin and phosphatidylethanolamin. The distribution for lysolecithin and sphingomyelin were no differences between normal and DAB-feeding rat liver. The distribution for phosphatidylcholin was elevated at 80th day of DAB-feeding rat liver than the normal one, and the distribution for phosphatidyl ethanolamin was bighn in DAB-feeding rat liver than the normal one at 20th day, but on the controry, it was changed at 40 and 60th day, and increased in normal than in DAB-feeding rat liver.

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Heinz bodies produced in vitro by X-irradiation or hydroxylamine hydrochloride were separated and collected. The lipids extracted from these Heinz bodies were analyzed as to their lipid and fatty acid composition by thin layer chromatography and gas-liquid chromatography. The purpose of the present experiment is to compare the composition of lipids and fatty acids in the Heinz bodies produced by the two different methods. In addition, the membranes of the untreated red cells were similarly treated and analyzed for comparison.
1. It was found that there were differences in the composition of lipids and fatty acids derived from Heinz bodies under different circumstances in which they were formed, even though they might appear morphologically identical on stained preparations. Namely, Heinz bodies contained a lipid fraction whose chemical composition was not necessarily the same and it was suggested that they were the degenerative products having the same characteristics.
2. Analysis of lipids of Heinz bodies indicated a marked decrease of the phosphatidyl ethanolamine ratio in comparison with the lipid fraction of the normal red cell membranes.
3. With respect to the fatty acid composition of Heinz bodies, there were a decrease of the stearic acid ratio and an increase of the oleic and linoleic acid ratios, resulting in a marked increase of the total unsaturated fatty acid ratio.
4. The development of X-irradiation anemia is due not only to an impaired erythropoiesis in the hematopoietic tissue but to an accelerated destruction of red cells in the peripheral blood. It is postulated that the peripheral red cells are rendered fragile by X-irradiation through an alteration of the lipid composition ratio of the cell membrane.

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Fatty acids extracted from Heinz bodies were added to Ehrlich ascites tumor cells and HeLa cells in tissue culture at various final concentrations of 0.2, 0.1, 0.05, 0.025%, and their effects on these cultured cells were morphologically observed.
It was shown that fatty acids derived from Heinz bodies exerted as nearly a potent cytotoxicity as OX substance on the cultured tumor cells. At the concentration of 0.2 and 0.1%, immediately following thier addition, almost all the cells showed marked degenerative changes such as bubbling and blister formation, lipid granulation in the cytoplasm and pyknosis of the nuclei and this was soon followed by cell floating and death. And even at the lower concentration of 0.05 and 0.025% the cell degeneration was observed in 3 hours and 6 hours respectively and became floating within 24 hours.
From these results, it is considered that the formation of Heinz bodies may be a biological reaction as the body defense mechanism,

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1. On the studies of the relatives of patients with SLE, a family was found in which two sisters were affected with SLE and other two showed positive tests for ANF. In another family, two were positive for ANF, other two families had one positive to ANF. In summery the number being positive for ANF was six (four females and two males).
2. It was emphasized on the basis of the results of various clinical and experimental conditions, that not only genetic factors but also functional unbalance among the genital glands, adrenortex and antibody-producing tissues participated in the onset of autoimmune diseases, especially SLE.
3. Serum γ and α₂ globulin levels were high in majority of SLE. In the course of treatment, there were two cases in which hypergammaglobulinemia became normal and beta-globulin was increased. This hyper-beta-globulinemia may indicate the increased complemental compoments in the serum.
4. All of clinically diagnosed 11 cases of SLE showed a positive LE cell test. The two-hour clotting method was the most sensitive and the cover-glass method followed. The LE factor had been shown to be 7S and 19S globulins by the direct fluorescent antibody technique.
5. ANF was positive in all cases of SLE, even those in remission. The most cases were showed to be heterogenous in all three immunoglobulin classes. IgG ANF was the highest in the titer and persisted for a long period though its titer gradually became low. On the contrary, IgM or IgA ANF (especially the latter) gradually diminished during remission. There were a poor correlation between fluorescent patterns and the types of nuclear antigens.

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1. There were no changes suggestive of SLE in the rabbits that had received hydralazine in daily dossage of 200mg per kilogram body weight for as long as 4 months.
2. Of the rabbits sensitized with egg white and, in addition, administered with hydralazine, many died suddenly after injections of egg white, but a few rabbits showed anemia, probably hemolytic in type. No LE cell phenomenon or antinuclear factor (ANF) was revealed in this group.
3. Slight leucopenia, albuminuria, hypergamma-globulinemia and increased IgG were found in the rabbits injected with rat anti-rabbit cell nuclei serum. Four rabbits showed nephritis, productive in form at necropsy, with deposition of γ-globulin in glomerular capil laries as revealed by the direct FAT. Weak LE cell phenomenon and ANF were positive in one rabbit.
4. It is presumable that, as in human cases, hereditary factor is impcrtant to produce the models of auto-immune diseases in animals.

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The regional lymph nodes of mice transplanted with Ehrlich ascites tumor have an inhibitory effect for cultured Ehrlich ascites tumor. The supernatant of homogenized regional lymph nodes were fractionated on Sephadex G-200 column chromatography. The elution curve was fractionated in main three peaks, and the later half of the first peak and second peak were effective for the suppression of Ehrlioh ascites cell growth. These two fractions were agreed electrophoretically protein.

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The supernatant of the regional lymph node of mice transplanted with Ehrlich ascites tumor were fractionated on DEAE Sephadex A-50 column chromatography with linear gradient elution. The eluted peaks were examined electrophretically. They were γ, β. α and albumin. β and α fractions were effective for suppression of Ehrlich ascites cell growth. Minimum effective β fraction was 300μg/ml. The sedimentation coefficient of β fraction was 6.5 S 20 w, and that of α fraction was 7.4 S 20 w.

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Disturhances of the cerebral blood flow were induced by compressing the tube connected to the carotid arteries or depressing the perfusion pressure by a modification of the cat brain perfusion method devised by Geiger et al. In these experiments the amount of cerebral blood flow was rapidly and drastically decreased and then restored to normal flow rate, and changes taking place in EEG during these experiments were studied. The results are briefly summarized as follows.
1. When the cerebral blood flow is rapidly and drastically decreased, within several to 10 odd seconds EEG turns to low voltage, then high voltage slow activities appear followed by flattening. When the cerebral blood flow is restored there appear bursts of high voltage slow activities at first, then fast activities gradually make their appearance, but the high voltage slow activities gradually diminish, finally returning to the original EEG These changes coincide well with those changes in EEG clinically observable as in the case of circulatory disturbance of the brain.
2. Aspects of the EEG changes differ according to the rate of decrease in the cerebral blood flow, and in the decrease of about 20-24% cerebral blood flow, EEG shows apparently no appreciable changes, but the frequency autoanalyzer reveales a slight decrease in the fast activities and an increase of slow activities.
3. In the instance where the cerebral blood flow is decreased 21-35%, EEG increases high voltage slow activities.
4. Where the cerebral blood flow decrease amounts to over 37% or it has transiently ceased, EEG takes the foregoing course and then is flattened.
5. Under the diminished conditions of cerebral blood flow, the decrease in the fast activities and the increase in the slow activities are found. The faster the EEG activity, the greater is its decrease, and the slower the EEG activity, the greater is its increase.
6. At the recovery stage of the cerebral blood flow, the rate of increase in the faster activities is high and slow activities are diminished
7. At the time when the cerebral blood flow is decreasing and also at the recovery stage when there appear high voltage slow activities, some of EEG's take the pattern of spike and wave complex.

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The brain was perfused with artificial blood containing [U-¹⁴C] glucose, and then induced the disturbances of cerebral blood flow as in the previous report. The changes in the glucose metabolism of the brain were studied by measuring oxigen consumption, carbon dioxide formation, glucose uptake and lactic acid output as well as the contents of glucose metabolites and their radioactivities. The results are presented as follows.
1. In the case where the cerebral blood flow is decreased 20% or less, the cerebral oxygen consumption maintains approximately a constant level; where the decrease is over 40% it decreases; and in the decrease of 20-30% both of the above phenomena can be observed.
2. As for carbon dioxide formation, likewise when the decrease is over 40%, the amount of carbon dioxide formed is decreased.
3. The glucose uptake is 0.44 μmole in average before the cerebral blood flow is decreased, it is 0.12 μmole 32-39 minutes after the cerebral blood flow is decreased, showing the decrease in proportion to the lapse of time. In addition, when the decrease of cerebral blood flow is over 40%, there is observed a decrease in the glucose uptake. With the lactic acid output there is observed no fixed tendency.
4. In the brain with blood flow disturbances the glycogen content is decreased to 1.65 μmole.
5. The lactic acid content of the brain shows a tendency to increase when the cerebral blood flow is low.
6. The relative specific activity of metabolites in the brain under the diminished blocd flow, is found to be glucose 100%; lactic acid 45%; and in comparison to 63-75% of lactic acid in standard brain perfusion, the decrease in the radioactivity of lactic acid is marked in the brain with blood flow disturbances. This means that the contribution of non radioactive endogenous substances to the glycolytic process has been potentiated as compared with that in normal state.
7. The relative specific activity of glutamic acid, aspartic acid, glutamine and respiratory carbon dioxide are 30%, 32%, 17% and 20% respectively. In the standard perfusion these values are high in the high level of EEG, respective values being 80%, 75%, 61% and 55%, and it is known that the lower the EEG level, the lower is such ratios. However, these values in the brain with blood flow disturbances, agree approximately with those values in the brain with low function.
8. The relative specific actiyity of γ-amino butyric acid is 14%, being lower than that in the standard brain perfusion where the brain function is low.
9. The relative specific activity of glycogen, while it shows a big standard deviation, is about 22%, giving a considerably higher value than what has been expected. Even from the decrease of the glycogen content in the brain, it can be surmised that the involvement of glycogen in the energy metabolism in the brain under the blood flow disturbance is hightened.
10. These results seem to indicate that in the glucose metabolism of the brain with blood flow disturbances, there occur general disturbances of glucose uptake, glycolytic pathway and the citric acid cycle.

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For the purpose to reveal the mechanism of the fatal injuries induced by X-ray the auther observed the biological effects of the compound-phospholipids from the irradiated animal, which proved to have a powerful hemolytic activity. Intravenous introduction of the phospholipids from the irradiated animal to the normal rabbit induced a severe anemia accom paning a marked leukemoid reaction, the degenerative changes of liver parenchymal cells and the hemorrhage in several organs, which are the main and important pathologic changes seen by the X-ray irradiation.
The findungs suggest that the cytolytic activity of the hemolytic lipids formed by X-ray irradiation is largely concerned to the tissue injuries seen after the irradiation.

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The literature dealing with the nature of irradiation disturbances is replete, but there seems to be no established theory on this phenomenon. It is needless to say that the elucidation of this condition requires further, extensive studies from biochemical, histological and functional aspects, because the irradiation disturbances are not by any means a single entity. Yamamoto of our laboratory states that the compound-phospholipids extracted from tissues of the rabbits exposed to total body irradiation possesses hemolytic and cytolytic effects similar to the effects of irradiation itself. In view of this, the present experiment was conduced in order to observe what effects such an extract would have on the growth of Ehrlich tumor culture cells (JTC 11, strain maintained in the laboratory of Cancer Institute, Okayama Univ.).
These cells were cultured in the media containing the aforementioned phospholipids extract in the concentration of 0.1% or 0.5%, and for the control the phospholipids extracted from non-irradiated rabbits was added to the medium in the same concentration. The percentage of degneration of Ehrlich tumor cells was calculated at intervals of one, 3, and 6 hours after the addition of the extract. Two days after the start of culture, the extract was added to the medium, and its effect on the proliferation of Ehrlich cells was studied on the fourth day of the addition of extract. Similarly, morphological changes of Ehrlich cells were observed by electron microscopy at the intervals of 3, 12, 24 and 48 hours after the addition of the extract.
As a result it was found that the phospholipids substance extracted from the irradiated rabbit showed a greater degenerative effect and stronger inhibition on the culture cells as well as it elicited more marked morphological changes than the extract from non-irradiated rabbit. From this, it is obvious that the extract of the irradiated rabbit has the effect to inhibit the cell growth and destroy the cells just as has been demonstrated in Part 1, which is similar to the disturbances brought about by irradiation.

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This experiment was aimed at finding out what time interval and dose of irradiation would the most useful inhibitory effects on experimental tumor. The tumor used was solid type Ehrlich carcinomas experimentally induced in mice, and the total dose of each irradiation was 5, 000R. The effect of irradiation was determined by the survival time of the tumor bearing animals (Table 1).
The interval of 72 hours which is thought to be most appropriate was taken as the standard of the interval. As a result it was found that the irradiation of 5, 000R at the interval of 72 hours with additional 100 R at every 24 hours lengthened the survival time of the tumorbearing animals to 4.0 days in average. The survival time of the controls given no irradiation was 20.4 days in average (Tables 2-4, Fig. 1). Further, in those groups given additional dose of 200 R or 300R every 24 hours, the length of survival days was rather shortened.
Besides these, some investigations were carried out to see what therapeutic effect such additional irradiation would have on the regional tumor and what systemic side-effects. The effect on the regional tumor was determined by the activity of succinic dehydrogenase system in the tumor and systemic irradiation damages in the blood picture (Tables 5-13, Figs. 2-6). The results revealed that in comparison to the group simply irradiated with 5, 000 R at interval of 72 hours, the group given additional 100 R showed the recovery of the succinic dehydrogenase activity to be inhibited but the blood picture did not exhibit any appreciable change. In the groups given additional dose of 200 R or 300 R the succinic dehydrogenase activity was diminished and also the damage to blood picture was in such a state that it could not be lightly taken of.
These findings indicate that when the irradiation is done at the interval of 72 hours and additional dose is given at the intermission period, such additional irradiation is effective at a certain appropriate dose. In this instance, the additional dose should be such as to inhabit the growth of tumor but not to cause much side-effect. In other woras, divided doses of irradiation should be within the range which would serve the very purpose of irradiation best.

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The total 23 cases were diagnosed as encephalomyelitis with abdominal disturbances in the Ibara districts from January, 1966 till August, 1967. Observations were made on incidences and characteristic clinical symptoms of the disease.
1. The number of the patienfs of adult females was greatest and more than twice as that of males. Onset of the disease was mostly found in the warm season. At present, particular age difference was not discernible in the incidences.
2. The patients with the disease were at first observed around 1965 and the number of those has a tendency to increase.
3. Characteristics of clinical manifestations were as follows:
(a) Abdominal pain and constipation were frequent as prodromal signs, whereas diarrhea was not frequent. The tender point at the abdominal wall was located approximately at D₁₀.
(b) As encephalomyelic nervous disorders at the lower limbs, a majority of the cases exhibited disturbance of foot flexion, tenderness and contraction of thigh adductor muscles. Ataxia, together with subjective and objective sensory disturbances as well as increased patellar reflex, was observed in all cases. By contrast, disorders at the upper limbs were not common. As optic nerve disturvances, about 40% of the cases showed weakness of vision, narrowness of visual field, etc.

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Drugs of adrenal corticosteroid and vitamine complex of B₁, B₆ and B₁₂ were therapeuti-cally administered on 21 cases out of the total 23 cases diagnosed as encephalomyelitis with abdominal disturbances in the Ibara districts from January, 1966 till August, 1967. Complementarity, administrations of antibiotics, ACTH, ATP, thioctic acid, etc. were added.
1. Treatment by adrenal corticosteroid improved both abdominal signs and nervous dis-orders in 48% of the cases. Such effects were also observed in cases showing recurrence.
2. Administration of vitamine B₁, B₆ and B₁₂ complex was effective to improve the nervous disorders, such as sensory or locomotor disturbances, in about 30% of the cases, although ineffective on the abdominal signs. The therapeutical effect of the vitamine complex, therefore, was lower than that of corticosteroid. However, in some cases showing no effect by corticosteroid treatment, symptoms were improved by administration of the vitamine complex. The complex was ineffective on cases showing recurrence.

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