Evidences have been accumulated that cell membranes in a frozen state are split and its interior structures are revealed. It has been proved by double replica method th at there is a unique cleavage along membranes and that convex faces, which are referred as the A face, and concave faces, the B face, are reciprocal ones of fractured membranes. Elevations and depressions revealed on one face by fracture through a frozen tissue are matched to those on the other, but there are little pits on the exposed membrane surfac es complementary to particles on the reciprocal ones, except for the junctional are as and certain plasma membranes. It might be due to difficulties inherent in this technique to reveal small and shallow indentations. Preliminary impression replica, relief of which i s perfectly matched to that of the fractured face, may be of value in studying origin of imperfect matching of freeze-cleave images on the reciprocal fracture faces.
Rat brain was perfused through the heart with 25 per cent glycerol i n saline solution and small blocks were dissected from the cerebral cortex. Fractured face of a frozen tissue block was replicated without shadowing using the modified freeze-etch appa ratus of Bullivant and Ames type. The replica was backed with silver and collodion film to enforce and distinguish the back of the replica. After dissolvement of tissue the replica was shadowed on the side of tissue, and collodion film and silver were succes sively removed by amyl acetate and nitric acid respectively.
On the preliminary impression replica, relief reverse to the usual fracture faces is shown and its resolution is nearly as good as that of ordinary ones. Tissue structures shown on the preliminary impression replica are not different from those see n on the freeze-cleave replica, except for surface characteristics.
On the freeze-cleave replica the surfac e of the fractured cytoplasm is granular and coarse on one hand, it appears smooth and fine on the preliminary impression replica on the other hand. There are particles on freeze-cleaved membranes and on the preliminary impression replica those are shown as pits, and very few particles are present on membranes, indicating that there are very few matched depressions on fractured membrane faces, for small depressions, if present on the fractured face, are clearly demonstrable by small protrusions on the preliminary impression replicas.
From the present experiments, it is shown that relief per fectly matching with fracture face is different from that seen on the freeze-cleave replica when relation of particles to depressions is taken into account. As the reciprocal freeze-cleave replicas prepared by double replica method, appear quite alike and indistinguishable from each other, it is obvious that reciprocal fracture faces are no longer perfectly matching though these are generated by single fracture. Since elements which lose complementarity on the fracture face are of the protein nature, such as particles on the membrane face and microtubules, probably intracytoplasmic proteins also, the loss of complementarity may be explained by plastic deformation of proteins at the time of fracture.

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In order to clarify the functional significance of the afferent average evoked potentials caused by sciatic nerve stimulation, their fundamental experiments were made by using rabbits.
1. As the result of the sciatic nerve stimulation, responses were observed in the following manner,3 negative responses (N₁, N₂ and N₃) 1 positive response (P) and 2 negative responses (N₄, N₅) in cerebral cortex.
2. The intravenous injections of barbiturate, chlorpromazine, myanesin, morphine and pentazocine, and the application of procaine on the peripheral nerve displayed an inhibitory action on such early components as N₁, N₂ and N₃.
3. By the application of the sciatic nerve stimu lation,3 small negative responses (HN₁, HN₂ and HN₃), followed by 2 large negative responses (HMI, HN₅) w ere observed in the hippocampus.
4. The pain ting of procaine on the peripheral nerve produced a strong inhibitory action on all the components of the afferent average evoked potential in the hippocampus.
5. The painting of procaine hardly acted on H wave and M wave of the spinal reflex,. but strongly inhibited the evoked muscular discharge by nociceptive reflex.
6. The lesion of VPM and VPL showed an inhibitory effect on th e early components of the afferent average evoked potential recorded from the cerebral cortex.
7. The lesion of VA, DM, VM and CM showed an inhibitory effect on such late components as P, N₄ and N₅.
8. The lesion of CG in hibited not only the early components of the afferent average evoked potential, but also their late components.
9. All the components of the affere nt average evoked potential in the hippocampus were inhibited by the lesion of VA, DM, VM, CM, VPM, VPL and CG.
As the results of the above various experiments, considerations h ave been given to the fact that N₁ and N₂ components of the afferent average evoked potential in the cerebral cortex appear after going up the afferent pathway which transmits a perception In, f rom the peripheral to the central, while N₄ and N₅ components appear via the central nuclei belonging to the diffuse thalamocortical projection system, and that these nuclei are also concerned with the appearance of the afferent average evoked potential in the hippocampus.

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The effects of CDP-choline on fetal rat lungs were investigated histologically and biochemically.
The first experiment was carried out in fetal rats to investigate the incorporation of CDP-choline-1.2-¹⁴C into pulmonary lecithin. Radioactive CDP-choline was injected int o each fetus in the right uterine horn of 19 days gestation intraperitoneally through the uterine wall, while their littermates in the left uterine horn were not treated as control. The fetuses, being delivered by cesarean section at 6,18,24,36, and 48 hours after the injection, were tested on the incorporation of radioactive CDP-choline into the rat lungs.
It was found that radioactivity of the pulmonary phospholipids of the fetuses in the right horn reached a peak 24 hours after the injection, and 0.31% of the given CDPcholine was incorporated. The radioactivity of the control group detected was very low,0.003% of the given CDP-choline. Radio thin layer chromatogram showed the spec ific radioactivity on lecithin fraction.
In the second experimen t, the effects of a single intraperitoneal injection of CDPcholine into the fetuses were studied. After laparatomy on the pregnant rats of 18,19and 20 days gestation, each fetus in the right uterine horn was administered intraperitoneally with CDP-choline (about 200 mg/kg), while a similar amount of saline was administered into each fetus in the left horn and served as control. The fetuses were delivered by cesarean section 24 hours later.
Pulmonary lecithin levels of the CDP -choline injected group were found to be little higher than those of the control group, and the number of granules in the pulmonary epithelial lining cells and lung maturation of both groups histologically showed no significant difference. The gas chromatographyic analysis indicated that lecithin was mainly composed of palmitic acid and no significant difference Was noted between both groups. Pulmonary palmitic acid levels of 21 days gestation were found to be significantly higher than those of 19 days gestation.
It can be concluded from th ese results that a single administration of CDP-choline to the fetal rat is not capable of accelerating the production of pulmonary lecithin of the lungs.

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Perinatal anoxia was experimentally produced in fetal rats to examine the effect of anoxia on pulmonary lecithin levels. The right uterus of adult Sprague-Dawley rats was exposed by laparatomy on the 19th day of pregnancy, and the right uterine artery was compressed for 2 minutes and then released for 1 minute. This procedure was repeated five times. Thus the rat fetuses in the right uterine horn experienced anoxia for a total of 10 minutes, while the littermates in the left horn were free from anoxia. The fetuses were delivered by cesarean section 24 hours after these treatments, and the lecithin content of the lungs was measured.
Pulmonary lecithin levels of the anoxic group were found to be significantly lower than those of the control group. Furthermore, it was found that body weight was significantly less and lung wet weight was significantly greater in the anoxic group. The lungs of the anoxic group showed partial atelectasis, interstitial edema, congestion and the presence of red cells and desquamated epithelial cells in the alveolar lumen.
A second experiment was conducted to examine the effects of CDP-ch oline and prednisolone upon the pulmonary lecithin levels of the anoxic fetuses. The fetuses in both uterine horns were exposed to anoxia by the same method used in the first experiment. The rats were divided into three groups. The fetuses in the right horn of the first group were injected intraperitoneally with CDP-choline (200mg/kg), the fetuses in the right horn of the second group with CDP-choline (200 mg/kg) and prednisolone (60mg/kg), and the fetuses in the right horn of the third group with prednisolone (30 mg/kg). The fetuses in the left horn of each group were injected with an amount of saline solution equivalent to the amount of medication received by the fetuses of the right horn, and served as the control group.
It was found that when CDP-ch oline was administered in this way (group one), there was no acceleration in the production of lecithin. But the newborn rats which had been injected with prednisolone (group three) showed a significantly higher titer of pulmonary lecithin than did the control group. As for group two, injection with a large dose of prednisolone did not result in an increase of lecithin production, but there was a significant weight loss.
It can be conc luded from these results that an anoxic episode during the perinatal period inhibits production of pulmonary surfactant, and is one of the causes of intrauterine growth retardation. The effects of anoxia cannot be corrected by a single administration of CDP-choline. An adequate prednisolone injection can accelerate pulmonary lecithin production, as already reported by other investigators. An overdose of prednisolone reduces the pulmonary lecithin levels, and retards weight gains of rat fetuses significantly.

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A neuropharmacological study was made in an attempt to clarify the relationship between the higher central nervous function and cerebral circulation, especially taking notice of muscular discharge, by using the Doppler ultrasonic flowmeter and an electromagnetic flowmeter.
The resu lts obtained were as follows.
1) Blood flow velocity was d etermined to be a fixed quantity through the use of the probe of a Doppler ultrasonic flowmeter which I specially adapted for this experiment.
2) In the case of the arousal reaction and the recruiting response, the effect upon the cerebral blood flow velocity and cerebral blood flow volume increased in the case of the arousal reaction, and decreased or did not change at all in the case of the recruiting response.
3) It wa s made clear that the cerebral blood flow velocity and the cerebral blood flow volume increased not only in the natural spontaneous state, but also in the case of the arousal or the recruiting response during the appearance of muscular discharge.
4) It was determined that the Doppler ultrasonic flowmeter was more sensitive to the cardiovascular dynamics than an electromagnetic flowmeter, when studied from the standpoint of the effects of therapeutic agents on the cerebral blood flow in cases of brain injury.
5) C oncerning the action of therapeutic agents in cases of brain injury, such as ATP, Cytochrome C, CDP-choline and so on, these increases of the cerebral blood flow volume were not observable in so far as no muscular discharge was revealed.
6) Following the administration of phenoxybenzamine and propranolol, it was demonstrated that the increase of the cerebral blood flow volume was related with the a-receptor when the evoked muscular discharge was obtained through the use of an electric stimulation on the midbrain reticular formation.
7) Following the studies of the momentary, instantaneous cerebral blood flow volume accompanied with an electric stimulation of the midbrain reticular formation or nucleus ventralis anterior in the thalamus, the influence of the evoked muscular discharge was most remarkable in the external carotid artery followed secondarily in the vertebral artery, while the tendency of the increased of the cerebral blood flow volume on the internal carotid artery was least detectable.
8) It was noted that the mom entary, instantaneous cerebral blood flow volume by the stimulation of the midbrain reticular formation or nucleus ventralis anterior increased to the maximum within 5 seconds after stimuli and decreased gradually thereafter.
Additional study was made concerning the alteration on the R-R interval of ECG. Also, the systemic blood pressure was examined and the increase of the cerebral blood flow volume indicated by the appearance of muscular discharge was discovered to be related to the increase of 1 cardiac stroke volume.

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The present experimental study is designed to investigate the effect of massive dosages of corticosteroid on the cerebral function electrophysiologically in rabbit by means of electroencephalography and electromyography under normal hemodynamic condition and under hemorrhagic shock.
The shock was pro duced by means of an induced hemorrhage; the mean arterial blood pressure during the induced hemorrhagic shock was maintained at 50 mmHg.
A massive dosage of corticosteroid has an inhibiting effect on the cerebral function under normal hemodynamics, whereas it has facilitating effect on the cerebral function under hemorrhagic shock. As concerns when to administer corticosteroid, its administration after the hemorrhage was more effective than before the hemorrhage. However, corticosteroid administered after the critical point, point where the depressed cerebral function on electroencephalogram is demonstrated, is not effective. The critical point of the depressed cerebral function on the electroencephaloglam was observed about 120minutes after the fall of the mean arterial blood pressure to 50 mmHg and the threshold value rose up to 200 per cent. After this critical point comes the irreversible shock phase.
It may be correct to say that the effect on the brain of corticosteroid administered in shock could be opposite to that of corticosteroid administered in circulatory normalcy, and that it might effectively accelerate the central nervous function.

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In recent years, venous grafts or synthetic grafts are the most commonly used substitutes for small peripheral arteries, particularly those less than 4 mm in diameter. However, the result with these prostheses have not been satisfactory due to frequent thromboses and inadequate flow.
From the previous experim ental or clinical works on reconstruction of peripheral arteries by arterial or venous allografts, it was evident that these grafts were often occluded by early thrombosis.
In recent years, cathode ray irradiated allografts provided by a Van de Graaff generator have been used in the transplantation of abdominal aorta, bones, peripheral nerves, etc, because of adequate sterilization and reduced foreign body reaction.
The purpose of this report is to describe the use of cathode ray irrad iated arterial allografts on the reconstruction of small peripheral arteries.
The following experiments were conducted:
1. The fate of irradiated arterial allografts implanted subcutaneously in rats was compared histologically with that of allografts which were fresh, frozen or stor e d in 70% alcohol.
2. Follow-up studies were made of 10 dogs in which irradiated arterial allografts had been implanted in the femoral artery to evaluate
i) long term patency by arteriography,
ii) gross and histologic ch anges of the implanted allografts,
iii) the luminal surface of the implanted allografts by scanning electron microscopic studies.
3. For the purpose of antigenic studies, extracts of canine arteries, irradiated, frozen and stored in 70% alcohol, were prepared and tested by the following immunolog i cal methods; precipitin, Ouchterlony gel diffusion, passive cutaneous anaphylaxis.
From the results obtained in these experiments, it can be concluded that irradiated arterial allografts are the most suitable substitute for the reconstruction of small arteries because of the paucity of morphological damage and inflammatory reaction compared with fresh or frozen grafts and those stored in 70% alcohol. The excellent and long term patency of the canine femoral artery implanted with irradiated arterial allografts was proved. On the basis of immunological studies, it is evident that cathode ray irradiation to the arterial allografts can greatly reduce their antigenicity.

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Reported: Electromyograms (EMGs) of 18 cases of SMON during the stationary stage were analysed quantitatively, i. e. their ampltude (AMH) and interval histogram (INH)were made using an ATAC 501-20 type computer. These histograms were compared with those of normal controls.
1) The AMHs and INHs of the normal controls showed their respective regular hyperbolical and bowl-shaped curves CN- (Normal) typeD, while the AMHs and INHs of the SMONs showed irregular curves with several peaks, classified by the author as the S-(Spinal cord), P-(Peripheral), M-(Muscular), and C-(combined) types. Among the se types, the P-type appeared most frequently.
2) In most cases, these finding s corresponded well with both the clinical picture and the routine EMG.
3) In som e cases, the routine EMGs remained within normal although the AMHs and INHs were abnormal.
4) Follow up: One or two years after onset, the patients were again given both clinical and EMG examinations. In one case, the EMG had clearly improved from a S-type to a P-type. On the other hand, a slight case, who had been at the stationary stage during the second year after onset, still complained of numbness on her lower extremities, and her EMG histograms showed a slight P-typs lesion and no remarkable differences from the histograms taken one year before. On the whole, almost all of the cases were improving very slowly.
5) EMGs of patients complaining of numbness: As almost all of the SMON patients complained of numbness of the extremities during the stationary stage, EMG histograms were taken of hysteria and thinner poisoning cases complaining of numbness on the extremities and compared with those of SMON cases.: The histograms of the hysteria cases were N-type and those of the thinner poisoning cases were P-type like those of the SMON cases.
Discus sed: The conclusion is that EMG analysis can discover organic damages much more efficiently by the author's method than by the routine method.

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Reported: Analyses of the EMG activities in diabetic patients were done with an ATAC 501-20 type computer and compared with the neurological findings in those patients by means of a grading system. The resnits were as follows:
1) Every diabetic patient was found to have a more or less abnormal EMG; even a patient displaying no neuropathy or no remarkable change on a routine EMG would present abnormal patterns on his amplitude and interval histograms.
2) The muscle groups affected were in the following frequency order: M. gastrocnemius)M. quadriceps femoris>M. tibialis ant.>M. biceps brachii>M. triceps brachii.>M. flexor carpi. ul.>Ist M. interossei.
3) The types of the histogr ams, obtained by author's method, were in the following frequency order: P-type>N-type>P+M-type>M-type.
4) Some correla tions were observed between the EMGs and clinical neurological findings.
5) The sum total of the grades given on the amplitude and interval histograms and those given on the neurological signs appeared to be a reasonable parameter for the severity of neurological disorders in diabetic patients.
Although affected by blood sugar level and disease duration, the correlation between diabetic neuropathy and aging was found to be high, especially over the age of fifty.

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Reported: Three cases of the Neuro-Behget Syndrome: One of the cerebro-brainstemspinal type (a 25 year old male) and two of the cerebro-brainstem-cerebellar type (a 35year old male and a 42 year old female).
Statistically observed (on 190 cases appearing in the literature plus our own 3 cases): This syndrome is much more common in males than in females, and onset usually occurs during the third or fourth decads. Onset is seen to be acute or subacute with fever and some of the triad of the syndrome. The initial signs are headche, spastic paraplegia, mental deterioration, and sensory impairment. During course, the following symptoms and signs appear successively: the brain stem syndrome, emotional disturbance, and sphincter disorder as well as meningeal, extrapyramidal, and cerebellar signs. Such a variety of clinical signs seems to be one of the characteristics of this disease. Motor disturbances, especially sever spastic paraplegia with laterality are common, but sensory disturbances are rare and occur only during the exacerbated phase. Most cases display alternate remission and relapse but some cases prove consistently progressive.
The direct causes of the death, confirmed by autopsy, are bu lbar palsy, infection, cerebral damage, heart insufficiency, and bowel perforation; and the most significant abnormalities, found by laboratory examination, are those of the cerebrospinal fluid and the EEGs.
The conclusion is that for the differential diagnosis of etiologically unknown neurological disease, the Neuro-Behget Syndrome should always be considered.

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