1) An attempt was made to study variation of cholinesterase and free nitrogen of aminoacids at globus pallidus lesion produced by various methods, such as oil-wax-, etopalin-injections and electrical coagulation. Eight normal cats for each these three groups were utilized in this study.
2) Both cholinesterase and free nitrogen of aminoacids were decreased in value for the first postoperative week in case of the oil-wax injection. They would, however, both come back to the normal values within the third postoperative week.
3) In case of the etopalin injection, the values of both cholinesterase and free nitrogen of aminoacids were considerably decreased for the first postoperative week and would not come back to the normal values within the third postoperative week.
4) The electrically coagulated lesion manifested the same tendency of the variation of the two substances as the oil-wax method for the first week. However, the recovery is comparatively sharp in contrast to the latter.

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1) This study was carried out for the histological analysis of the lesions produced by the methods of oil-wax-, etopalin-injections and electrical coagulation.
2) Etopalin tends to produce an irregular and large degenerative necrosis at the perifocal area of the lesion. This is, therefore, not desirable technique for the purpose of stereotaxic surgery to make a small, localized lesion.
3) In case of the electrical coagulation, the lesion reveals an extensive reaction of the tissues and could be hardly expected to make as large as it will be expected in size.
4) Although the operation is difficult at the time of the injection, oil-wax can produce a necrotic lesion as the result of slow tissue reaction against the expanding pressure, the mildest lesion-making process can be expected.
5) In general, Etopalin manifests the severest reaction around the contact tissue and, in contrast, the mildest can be obtained in case of the oil-wax technique.

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1) In this study, cats were utilized to make a histochemical study of dehydroxyrase at globus pallidus lesion produced by the methods of oil-wax-, etopalin-injections and electrical coagulation.
2) In case of oil-wax-produced lesion, the immediately contact tissue is negative or slightly positive in the reaction of dehydroxyrase and, in contrast, the outer layer is normal in the value, showing a normal aerobic glycolysis.
3) The Etopalin method manifested a weak reaction or hypofunction of aerobic glycolysis in the comparatively extensive area around the absolutely necrotized contact border tissue.
4) A normal reaction is obtained at the tissue near the contact border tissue with a coagulating electrode, maintaining the sufficient aerobic glycolysis.

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It became recently known that there are two types of characteristic spikes in EMG at voluntary contruction of skeletal muscle; the one is of low voltage and long duration, and the other, of high voltage and short duration. In this study a detailed observation was done for analysis of these spikes.
1) They would not be changed in duration by alternation of the recording electrode position.
2) In the histograms of the voltage and duration of the spikes recorded separately from flexor digitorum profudus muscle and soleus muscle, it was revealed that the one from the first muscle was of 50-550 μV in voltage range with two peaks at 100-200 μV and 300-450 μV aud of 0.5-5.5 ms in duration range with also two peaks at 1.0-1.5 ms and 2.0-3.0 ms. The one from the latter muscle was ranged in similar manner as in the first, but the peak was found solely at 100-150 μV in voltage and 2.5-3.5 ms in duration.
3) In another figure drawn both voltage and duration, it is interesting to know that the first group was subgrouped into two; the one is of short duration and high voltage and the other is of long duration and low voltage. In contrast to it, the latter was of low voltage and long duration.
4) In the -s curve based on average discharge interval and standard deviation of its irregular variation of these spikes, the one of high voltage and short duration manifested a sharp increment aud the other of low voltage and long duration, a slow increment.
5) Two types of such characteristic diphasi spikes were also obtained in M wave of provocation EMG. The latency appeared to be short in the spike of high voltage and short duration compared with the other.
6) The reflactory periods, both in absolute and relative, were also found to be short in the spike of high voltage and short duration incomparison with the other.
7) It is to be noted that the intermediate spike between these two types was found, although relatively in few incidence, both at voluntary contruction and in M wave at provocation.
It seem to be concluded that the spike of high voltage aud short duration corresponds with the one of kinetic NMU and the other, with the one of tonic NMU.

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The authors have experimented to detect the planktons from the drowned dead bodies under the special conditions. Rabbits were thrown into the ditch water after they were injured the caro id artery, during the terminal breathing by strangulation and concussion of the brain. It was tried to detect the planktons after the incinaration of organs and bones. Results are following;
1) Planktons could not be detected from organs and bones of rabbits that were put the ditch water into stomach with catheterization.
2) Planktons could be detected from the organs and bones of rabbits that were drowned to death after injured to the carotid artery, but longer the time after injured, lesser the planktons can be detected.
3) Excepting lung, there were no planktons in the organs and bones of the rabbits that were thrown into the river after death by bleeding.
4) From the organs and bones of the rabbits that were thrown into the river under standstill of therminal breathing by strangulation, several planktons could be detected though in small number.
5) Planktons could be detected from organs and bones of the rabbits that were drowned to death in the river under concussion of the brain.

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The authors have experimented to detect the vegetative planktons (diatoms) from organs and bones of the rabbits that were drowned to death under light and deep narcosis with chloroform, ether and Ravonal, under light, middle and deep drunkenness, under hypnosis by administration with Brovarin, Adorm and Valamin. Results are following;
1) Planktons could be detected from organs and bones of rabbits that were drowned to death under narcosis with chloroform, ether and Ravonal, under few degrees of drunkenness with alcohol, under hypnosis by administration with Brovarin, Adorm and Valamin.
2) Planktons could be detected almost same number in the experiments using the rabbits under narcosis with ether and Rabonal and light narcosis with chloroform, but smaller numbers under deep narcosis with chloroform.
3) As become deeper the drunkenness, smaller numbers of planktons could be detected in the experiments used alcohol.
4) In the experiments used rabbits under hypnosis by administration with Brovarin, Adorm and Valamin, total number of planktons detected from the organs and bones of rabbits lesser in case of using Brovarin, Adorm and Valamin orderly.

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An attempt was made to make a study on an incorporation of GABA with brain when it was administered to a mouse abdominal cavity.
1) When 0.5 mg per body gram weight of GABA was administered, the content in the mouse brain was not changed, although the one in the liver was rapidly and yet temporarily increa- sed.
2) A one week successive administration, 0.3 mg per body gram weight of daily dosage could not make any change of the content in the brain.
3) A study with ¹⁴C-GABA revealed a rapid temporary incorporation with the liver and the kidney and, in contrast, considerbly little with the brain.

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Studies were carried out to clearfy GABA metabolism in the brains of an epileptic, the latent cerebral local anaphylactic (L.C.L.A.) rabbit and ep-mouse administered with ¹⁴C-GABA.
1) Incubated with a brain slice, radioactivity of ¹⁴C-GABA was slightly detected in glutamic acid, aspartic acid and glutamine in the incubation medium.
2) In this case, most of the radioactive carbon of GABA was converted into ¹⁴CO₂.
3) The epileptic brain appeared to decreased in the ¹⁴CO₂-activity, compared with the non-epileptic. In case of the epileptic brain, the activity in the focus was lower than the one in the non-focus.
4) The study with L.C.L.A. rabbit and ep-mouse could not manifest any difference in GABA metabolism in contrast to the normal control groups.

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A genuine epileptic and experimental animals such as L.C.L.A. rabbit, ep-mouse and vitamin B₆ deficient mouse were utilized for this study to measure the transamilase activity.
1) The focus in the epileptic brain manifested the decrease of the transamilase activity compared with the non-focus and also the non epileptic.
2) There was, however, no significant difference between the transamilase activities of the epileptic non-focus and of the non-epileptic.
3) In both cases of L.C.L.A. rabbit and ep-mouse, no difference could be found in the activity in contrast to the control groups.
4) The transamilase activity of the VB₆ deficient mouse brain was seemed to be considerably decreased in contrast to the control.

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1) An attempt was made to analyse the pre- and postoperative EMG in order to correlate the physiological mechanism of pyramidal and extrapyramidal systems with clinical symptoms in course of stereotaxic surgery.
2) Globus pallidus, thalamocapsular area and ventrolateral nuclei of thalamus were selected as the main anatomical locations for target.
3) The thalamocapsular lesion manifesting the shortening of the silent period of the Hwave recovery curve signifies the increased activity of the α -system and a symptom of pyramidal system disorders.
4) The prolongation of the period in the cases of pallidal and VL lesions represents the decrement of α-motoneuron excitability secondary to the functional drop of γ-system.
5) It seems to be clear the silent period shortening the lesions are involved in internal capsule and the prolonging ones are in the nuclei of the extrapyramidal system.

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1) Utilizing involuntary movement patients, GP, thalamus and their neibouring structure were electrically studied by depth electrodes, at rest and activated states. In addition to it, stretch reflex and H-wave of EMG at stimulation were analyzed.
2) Electroencephalography of GP reveals 10-12 cps, α-band, sinewave at rest. However, thalamus and TC represent faster waves of nothing characteristic in resting EEG. It is to be noted that these EEG manifestation would not be altered by stimulation.
3) Stimulation of GP causes increased stretch reflex, but no alternation in H-wave. Stimulation of TC manifests increased amplitude of H-wave without causing any change in stretch reflex. Nothing could be found at stimulation of thalamus.
4) It appeares to be concluded that TC, involving pyramidal tract in internal capsule, causes the increment of α-system excitability at stimulation and, in contrast, GP stimulation represents an increment of γ-system excitability.

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1) Electrical stimulation being applied on peripheral nerves of four extremities, tremor inducing test was carried out on parkinsonian patients.
2) Tremor can not be abolished by peripheral γ-block and can be induced in that state.
3) Tremor can be abolished by total peripheral nerve block and can not be induced in that state.
4) In general anaesthesia, tremor can neither be observed nor induced as in case of sleep.
5) Tremor does not have anything to do directly with γ-system. Tremor and rigidity are independent each other so far as their functional mechanisms are concerned.
6) The mechanism of tremor is based on spinal anterior horn cells activated by upper center.
7) Intermediate small discharge between grouping voltages of a muscle in tremor is simply a conducted field potential from the antagonist.

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In the present paper an autopsy case report of Neuro-Behçet's syndrome has been made and patho-histological studies of the brain of the 8 autopsy cases reported previously as well as those of the present case have been conducted.
The present case was a 47-years old man. In the ninth years after the onset of symptoms in the oral cavity and in the genitalia he was troubled repeatedly with diplopia. Gradually there appeared such symptoms as the disturbance of his gait, forced laughing, disturbance of speech, ataxia, and the inflammation at the sites of injection. He had remission and aggravation of these symptoms. However, he died of respiratory paralysis 17 years after the onset of oral aphtha and 8 years after the onset of diplopia.
In the patho-histological observation of this case there were seen perivascular cell infiltration covering the entire cerebral parenchyma as well as a portion of meninges, revealing a picture of chronic disseminated panmeningoencephalitis and cerebral abscess of medulla oblongata.
In addition, there were fibrinoid degeneration of the blood vessel wall and such degenerative changes in the blood vessels as fibrosis and proliferation of collagen fibers; and also circumscribed foci of glia cell proliferation, softening cavities and abnomal proliferation of consumptive pigments. These pathlogical changes were recognized in the entire brain, but they were especially prominent in brain-stem. There were no pathologica changes in the optic nerves and in the spinal cord.
In the investigations of the patho-histological picture of the brain in our case as well as in the cases reported in literature, the common findings in Neuro-Behçet's syndrome are extensive disseminated perivascular cell infiltration of the brain, softning accompanied by a moderate proliferation of microglia cells and degenerative changes of the blood vessel wall. while these pathological changes are especially marked in the brain-stem, to a greater degree in the white matter than in the grey matter.
Neuro-Behçet's syndrome has been found to resemble periarteritis nodosa and clinically it shows many common pathologic changes as found in collagene disease. Hence, it is assumed that Behcet's syndrome belongs to the collagene disease. (Author's Abstract)

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With the purpose of investigation on changes in growth pattern, bone marrow tissue culture was conducted at various intervals in leukemic patients who finally arrived at remission by antileukemic treatment. The growth pattern specific to leukemia was observed to change as the patients impro ed by the treatment. Further, the relationship between the changes in the growth pattern, and the various signs and symptoms and life span was studied.
1. The growth patterns at remission were classified into those four types according to the grade of the changes: leukemic, intermediate, normal and hypoplastic patterns.
2. It was demonstrated as the most effective way that acute leukemia was treated until the bone marrow growth pattern changed into the normal pattern and chronic leukemia until it turned into the hypolast c pattern.

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The blood cells in the bone marrow of leukemia before and after antileukemic chemotherapy and those of hypoplastic anemia were observed morphologically by vital staining.
1. In acute myelogenous leukemia at remission, the mitochondria of the atypical promyelocytes displayed variation in size and those of the lymphocytes and the erythroblasts showed the same change. Some of neutral red granules of the young neutrophilic cells became larger in size.
2. In acute lymphocytic leuklemia at remission, the mitochondria of the lymphoblasts and the atypical lymphocytes exhibited a considerable variation in size and during the tissue culture, some of those mitochondria developed small vacuoles.
3. In monocytic leukemia at remission, the mitochondria of the promonocytes displayed a slight variation in size.
4. In chronic myelogenous at remission, the mitochondria of the young neutrophilic blood cells became varying in size and many neutral red vacuoles appeared in the cytoplasm of mature neutrophils. Appearance of many unstainable vacuoles around the nucleus of the myslocytes seemed to be characteristic in the chronic myelogenous leukemia at remission that showed the hypolastic pattern in bone marrow tissue culture.
5. In hypoplastic anemia, unstainable vacuoles which were observed in treated chronic mylogenous leukemia made no appearance.
In conclusion, the morphologic changes observed after treatment were considered to be the pictures of cellular degeneration that were superimposed over the cytologic characteristics of leukemic cells and accordingly, by this feature leukemia was able to be cytologically diagnosed even at remission.

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The behavior of the marrow megakaryocytes of leukemias before and after chemotherapy was observed by bone marrow tissue culture technic. Steroid hormone was used in acute leukemia and myleran in chronic for antileukemic chemotherapy.
1. Before treatment, the megakaryocyte count was less than normal level in acute leukemia and on the contrary, greater in chronic. The megakaryocyte function decreased both in acute and chronic leukmias.
2. After treatment, the megakaryocyte count was found markedly increased in chronic leukemia and frequently above normal level in acute leukemia. The megakaryocyte function also increased towards normal level both in acute and chronic leukemias and the increase was more remarkable in acute leukemia than chronic.
3. At relapse of acute lymphocytic and monocytic leukemias, both the megakaryocyte count and function showed no remarkable changes.
4. On the relation to the growth pattern, in acute leukemia, both the megakaryocyte count and function showed the tendency of decrease when the growth pattern proceeded from the intermediate pattern to the normal. In chronic leukemia, megakaryocyte function increased moderately and the count decreased markedly when the pattern proceeded from the normal pattern to the hypoplastic.
From the results obtained and on the consideration of the hemorrhagic diathesis, it is advisable that in acute leukemia, antileukemic treatment should be given to maintain the normal pattern in bone marrow tissue culture and in chronic leukemia, administration of antileukemic agents is to be quitted as soon as the hypolastic pattern is observed, and after a careful observation is paid on the bone marrow tissue clture findings and clinical signs and symptoms, the administration should be started again when the culture returns to the normal pattern.

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In the first paper, the results of experiments are presented, in which an attempt was made to develope methods of cytoloyical observation and tissue culture techniques suitable for the morphological studies on Yoshida sarcoma cells in tissue culture.
In the present experiments it was established that the method of cell-suspension culture was the most suitable one for such studies. Advantages of this method of tissue culture are as follows: 1) It enables us to observe minute cell structures by phase-contrast microscopy; 2) it is easy to count cell number; 3) it is possible to evaluate cellular activity; and 4) it enables us to take out cells in culture easily and to conduct various cytological studies, such as routine staining, peroxidase reaction, supra-vital staining and carbon particle phagocytosis of these cells.
Next, conditions of the medium necessary for the the survival and growth of Yoshida sarcoma cells were investigated. It was found that the suitable conditions of the medium were, pH 7.2-7.4, glucose solution at the concentration of 150mg/dl, 44% Hank's solution, 19% chick embryo extract, 29% horse serum, 8% Yoshida sarcoma ascites (extract), 0.8mg/ml RNA, and 0.8γ/ml vitamin B₁₂.
The pretreatment of the medium with Yoshida sarcoma cells eliminated the problem of lag phase, and also an addition of a small portion of crushed chick embryo tissue (containing fibroblasts) improved the cellular growth. In order to keep cells floating, roller tubes were placed in an incubator kept at 37 C.
Yoshida sarcoma cells cultured in such a medium under the conditions afore-mentioned increased in number at an early stage but later their population decreased gradually. Observation was possible for a certain period of time, the maximum being 12 days.

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By means of cell-suspension culture as described in Part 1, morphological changes in Yoshida sarcoma cells were studied by phase-contrast microscopy during the cultivation.
Those Yoshida sarcoma cells in ascites as well as those immediately after the start of tissue culture were well delineated. Their cell margin swayed slightly and slowly like monocytes, but the cells themselves did not move at all. Most of the nuclei were kidney-shaped and some were lobulated. Nucleoles were ovoid in shape and usually these were one or two distinct ones. Their nuclear membranes were likewise clearly visible.
At the recess of the nucleus there could be recognized a Golgi field on both sides of which there were mitochondria that exhibited slight motility. Cell mitosis could be observed in about 2 to 4 per cent of them.
After one day of culture, there was an increase in the number of cells and mitotic figures but no marked morphological chaeges were obseved.
After 4 days, the nuclei became lobulated or irregular in shape, nucleoles grew indistinct, mitochondria turned granular, and vacuolization of Golgi field occurred.
After eight days, the cells that had been swollen by the 7th day were completely destroyed, and among these disintegrated cells there were a few, scattered, new cell with high celluar activity.
After 12 days, even these newly-fosmed cells were mostly destroyed and only a few survived.
When these few surviving cells together with the culture medium were injected into the abdominal cavity of rats, 11 days later the animals developed ascites containing many Yoshida sarcoma cells.
Apart from these experiments, various other cytological studies were carried out during the various stages of tissue culture and particularly fluorescence microscopy provided unique findings on the cultured cells.

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In this paper are presented the results of experiments conducted to see the influences of 5 different anticancer agents (Colsemid, Carcinophillin, Glucosamin, Nitromin and OX substance i.e. unsaturated fatty acid fraction extracted from the liver of rabbits given total body irradiation) on cellurlar morphology, activity and cell count of Yoshida sarcoma cells in cell-suspension culture. These are summarized as follows:
1. In the case with Colsemid (dose of 10 and 100γ/ml, incubated for 12 hours): After 12 hours, granulation of mitochondria and an increase in the number of pair cells were observed. Cellular damages were greater with the dose of 10 than with 100γ.
2. In the case with Carcinophillin (dose of 50 and 100 units/ml, incubated for 12 and 24 hours): There was marked shrinkage of the cells; shrinkage of the nuclei, nucloles and cytoplasm, with an increase in the number of shrunken, degenerative granules. Despite such a high degree of cellular damages there was no significant decrease in the number of cells. It may have been due to the fact that the shrunken cells are hard to be destroyed.
3. In the case with Glncosamin (dose of 1 and 10 mg/ml, incubated for 24 hours): In the dose of 1 mg/ml of Glucosamin no marked damage of the cells was recongnized. However, with the dose of 10 mg/ml, 12 hours after incubation, the cells presented an entirely different picture as compared with the case of 1 mg/ml. The cells were filled with many round, solid-looking vacuoles, and the nucleus and mitochondria were barely visible among these vacuoles. After 24-hour of incubation, masses of vacuoles and degenerative granules completely replaced the cells.
4. In the case with Nitromin (dose of 100 and 500γ/ml, incubated for 12 and 24 hours): After 12 hours, swelling of the cells was marked, and the general struture of the cells appeared indistinct. Some of the cells had non-motile, peculiar pseudopodia. A few so soft-looking vacuoles were recognized. Mitochondria became granular and further they were transformed into small vacuoles.
After 24 hours, only aggregations of large and small vacuoles were seen floating instead of live cells.
5. In the case with OX substance (dose of 0.1 and 0.4 ml/ml, incubated for 6, 24 and 48 hours): After 6 or 24 hours of incubation, there could be recognized shrinkage and. disintegration of the nuclei and lipoid granules, or swelling and disintegration of the cell margin. These changes were quite similar to the simultaneous effects of Carcinophyllin and Nitromin upon the cells.
In the case with 0.4 ml/ml of OX, after 24 hours of incubation, the cells were completely destroyed, and with 0.1 ml/ml of OX, after the same period of incubation afterwads the majority of the cells were dead, leaving a few surving cells among the dead ones.
In the case where the medium was supplemented with unsaturated fatty acid fraction extracted from the liver of rabbits not irradiated with X-rays, the cell growth was greatly enhanced and from the viewpoint of cellular morphology and cell count, there could be obseved no injurious effects of this fraction on the cells.

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