The attempt to type Coxsackie viruses isolated from the fecal specimens of cases with Guillain-Barré syndrome had disclosed relevant two strains Dohi (A-19) and Okumoto (A-2) in 1952. Further studies were made on 7 cases in 1953. Three strains Inaba (A-2), Kamada (A-2+A-5) and Saragai (B-1) were obtained in suckling mice. Pathogenicity of Inaba and Kamada strains revealed flaccid paralysis up to mice 10 days old with the finding of the generalized involvement of muscle. In contrast, Saragai strain exhibited spastic paralysis and tremor together with occasional cramp or jaundice up to mice 4 days old, and pathology revealed the involvement of fat tissue, pancreas, CNS, liver and muscle respectively.
Seven paired sera were concomitantly challenged with Inaba, Kamada, Kamada 2 (A-5), and Saragai strains in a measure of neutralization and complement fixation. Case 2 showed a rise in neutralizing antibody levels against A-2, and case 3 & 7 had the rise in B-1 titer. But case 1, 4 & 7 revealed a high titer against A-2 in the initial serum and a specimen taken in the convalescent stage showed the same titer. Case 5 & 6 being twin brothers of age 14 were with negative reading in titer. By and large, C-F antibodies appeared earlier in the acute sera and fall in the convalescent stadium. But in the cases with negative virus isolation, except for case 7 in Saragai strain, it seemed that the correlation of the results of C-F tests with those of the neutralization tests was obscure.
The significance of these agents in relation to the Guillain-Barré syndrome abundantly encountered in Okayama prefecture during 1952-53 would remain to await further investigation.

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Evidence of double infection of two discernible types of group A Coxsackie virus has been obtained from a case with typical Guillain-Barré syndrome associated with eruptions. The onset of the girl of age 4 years was characterized by the flaccid paralysis of both legs. Four days later the measles-like eruptions appeared on upper arm which covered whole upper trunk within following 48 hours.
Isolation of viruses was scheduled from the fecal specimens of 7 days after onset by the method developed by Beeman et at. with the positive results. Typing of isolated this Kamada srtain with Coxsackie virus group A type 1-4 and Kudo (A-5) strains revealed the puzzling double character of A-2 and Kudo type. To determine whether this is the case of mixed infection or of undisclosed entity the separation of the viruses was accordingly performed by the method described by Huebner et al.
The mixing virus type at 5th passage level was reciprocally eliminated by the addition of corresponding single antiserum, whereas two pure types were independently identified as A-2 and A-5. Similar attemts from the original fecal specimens were repeatedly successful. Further cross-neutralization test was made with Inaba, Kamada-2 (5th passage suspension derived from 10^-3 Kamada strain incubated with A-2 antiserum), Kudo and H-1 (A-5) strains.
The rise in antibody titer of paired Kamada sera was demonstrated in the reading from 1; 10 to 1; 100 against A-2 and equally 1; 100 against A-5.
The significance of the relevant double infection in this case to the symptome observed has not been obviated yet.

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Although the use of antiserum, combined with a course of vaccine, has been recommended recently in the prophylaxis of rabies, the experimental basis of this procedure is not yet obtained.
In studies reported here, effects of passively injected antiserum on the immunizing activity of UV vaccine were examined in experimental rabies in mice.
Ultraviolet inactivated vaccine and rabbit antiserum were injected into mice with various intervals, then antibody levels in the blood and the resistance to intracerebral challenge of fixed virus were followed up to 4 weeks.
The resistance of mice injected with vaccine and antiserum simultaneously, were much lower than those of control mice injected vaccine alone. The production of antibody was also completely depressed.
Even when antiserum were introduced 5 days before vaccination, the development of both resistance and blood antibody were completedly depressed.
Antiserum injected 3 and 7 days after vaccination, could not alter the development of immunity by vaccination.
From these results, some considerations on the problems of human antirabies treatment were discussed.

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Up to now, only two viruses are known to produce hemolytis in vitro, mumps and Newcastle disease. However, during the course of studies on Hemagglutinating Virus of Japan (HVJ), previously known as Newborn Pneumonitis Virus or Sendai Virus, hemolytic activity of the virus on chicken or guinea pig cells has been noticed. As two schools in this country have already ovserved the same fact, this paper chiefly concerns to further explanation of this property, especially from the view-point that whether the differentiation between the phenomenon of elution and that of hemoysis is possible or not.
On the purification procedure of the virus, using adsorption to and elution from chicken red cells, both hemagglutinin and hemolytic activities detected in the infected allantoic fluid have been recovered in the same manner. Specific immune serum against the virus inhibited the hemolysis as well as hemagglutinin. These results may suggest that the hemolytic activity is intrinsic character of the virus itself and is not due to other constituents of the allantoic fluid.
The mediation of the hemolysis through the same receptors of red cells as are concerned with hemagglutination has been also confirmed by the fact that cells could be rendered insusceptible to hemolysis by treatment with enzyme RDE or by the action of influenza virus PR 8. The result described above will indicate that the adsorption of virus to red cell is an indispensable step for viral hemolysis to follow.
Further kinetic studies on the hemolysis after adsorption, revealed the difference between the phenomenom of elution and that of hemolysis. When cells on the course of elution at 37°C was chilled, the elution will not follow so far, but the hemolysis will continue in the same manner as when these cells were kept at 37°C. Moreover, when adsorbed cells were kept under different temperature, the characteristics of elution curve was quite different from the curve of hemolysis.
In final experiment, eluting ability, enzymatic activity and hemolytic activity of both formalin treated virus and purified virus with methanol have been compared with that of untreated virus. Methanol treated virus did agglutinate chicken red cells, eluted from cells as well as that of control. Moreover, the treated virus had full activity of enzyme to destroy α-inhibitor. Never the less, hemolytic activity has completely lost after methanol treatment. Thus the dissociation of luting activity and hemolytic activity has been established with treated virus. The difference of heat stability found between the hemolytic property and hemagglutinin was also in favour of this conclusion.
However, by means of ultraviolet irradiation or by treatment with sodium periodate, the hemolytic property of the virus was not differentated from the eluting activity.

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In order to obtain a potent RDE preparation, biological studies on the 4Z strain of V. cholerae, on production media and chemical studies on the purification procedure have been conducted. The conclusions will be summarized as follows:
1) Original strain of 4Z was plated out on nutrient agar and colony selections have been repeated. Characteristic S form colony which has been seleted out produced more potent RDE than RS or R type. Moreover, constant titer of RDE was always obtainable with the selected strain.
2) In order to find out the best production medium, 1% nutrient agar, 1% peptone water, meat extract peptone broth and meat infusion broth were examined. The former two was more favourable and among them peptone water was more convenient for further purification.
3) Different kind of peptone was tested for the production of RDE in peptone water, and with Neo-peptone and Teruuchi-peptone, highest titer was obtained.
4) The addition of hog stomach mucin to the peptone water increased the production of RDE as much as 20 times when compared to the plain peptone water. Optimum concentration of mucin was 15-25mg/ml and in this medium maximum titer was obtainable after 16-18 hours incubation.
5) Instead of mucin, egg-white was also available for the stimulation of production.
6) Purification procedure from culture filtrate was examined. Adsorption to and elution from chicken red cells was most important step for purification.

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Two hepatotropic viruses, i.e. mouse hepatitis virus isolated by Buescher' and Ectromelia virus, were inoculated into mice intraperineally and histopathological changes of mice were pursued with particular emphasis on the features of the liver.
In addition to these studies, the level of liver glycogen and blood sugar of mice was estimated day by day after infection. One of the hepatotoxic substance, toyocamycin, which has been isolated in our laboratory from a streptomyces filtrate, was also tested just for comparison. The results obtained in such studies are the subject of this communication and will be summarized as follows:
1). All of the mice injected with two different viruses and toyocamycin died almost within 72hrs. On the histopathological examination, conspicuous changes were chiefly limited to the liver. These changes were characteristic with each agent.
2) Both the suger content of the blood and liver glycogen decreased gradually after inoculation of the above two viruses. However, with toyocamycin injection, liver glycogen decreased without any detectable changes of blood sugar content.
3). When glucose tolerance test was employed, the drop down of reserve activity of liver glycogen have been recongnized with these three groups.
4). In glucose tolerance test, administration of dl-methionine or glucuronic acid, particularly the latter, was favourable for improving the liver function with toyocamycin injected mice, but no effect was observed with the mice infected with viruses.
From the above observation, the auther concluded that with viral hepatitis, low blood sugar content would be partly explainable with lowered activility of liver function, but the disorders in other parts of body, which would participate in controlling the blood sugar level should be also considered. On the other hand, toxicity of toyocamycin was rather limited to the liver.

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Characteristic lesions on the endodermal layer of the chorioallantois removed from chicken embryos inoculated with Sendai virus has been already reported. These changes were rather proliferative than degenerative, such as bridge formation between cells of endodermal lining. Later, similar pathological features were also found on the bronchial cell of infected mice.
Futher histopathological studies of the bronchial cell revealed inclusion bodies related to that described by Harford et al. with influenza virus. As Sendai virus is in particular position among the family of influenza virus, these cytoplasmic changes have been pursued both histopathologically and by electron microscopy with correlation to the viral growth in mouse lung, in order to get further information on the characteristics of this virus.
In sections stained with hematoxylin and eosin, these bodies usually appeared 3 days after infection. At this early stage, they were basophilic and shaped as round or ovoid, within a day, the inclusions were surrounded by envelopes of acidophilic material. Though the small basophilic segments were still detectable after enlargement of the latter acidophilic matrix, final features of eosibophilic inclusions had no inner structure and seemed to be just homogeneous. These mature bodies were almost as large as nucleus.
When these sections were stained by pyronine and methylgreen, basophilic part was stained by methylgreen and eosinophilic part by pyronin. However, mature bodies were Feulgen negative.
These characteristic bodies were not detectable by instillation of normal allantoic fluid. Moreover, serotherapy of infected mice by injection of anti-HVJ serum prevented the appearance of the body. On the other hand, treatment of mice with xerosin had no effect on the appearance of these bodies. In this case though the lung lesions have been fairly retarded, virus growth was not affected at all. Thus the detection of inclusion bodies were correlated to the viral growth, but not to the inflammatory changes.
Identification of inclusion bodies which have been demonstrated in ciliated cells of infected bronchial epithelium by electron microscopy with that found by optical microscopy was not studied in detail.
In summary, the characteristics of cytoplasmic inclusions which have been detected in the bronchial epithelium of mice infected with Sendai virus, are quite similar to that reported by Harford et al. with influenza virus, but the criticism of Morgan et al. on the latter work, must be also considered in this study.

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