Okayama Igakkai Zasshi (Journal of Okayama Medical Association)

STUDIES ON THE PREVALENCE OF JAPANESE B ENCEPHALITIS IN OKAYAMA AND KAGAWA PREFECTURE IN 1956 AND ON NEWLY ISOLATED STRAINS, “OKAYAMA 56 E” AND “OKAYAMA 56 F”

Masuo Ogata, Hisanori Okuda

1956 Volume 68 Issue 11supplement Pages 1-5
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_1

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In the prevalence of Japanease B Encephalitis in Okayama and Kagawa Prefecture in 1956, the number of the patients increased with a sudden drop in temperature by typhoon No. 9.
In the past, the Japanease B Encephalitis spreaded in order to Kagawa Prefecture→south Okayama Prefecture→north Okayama Prefecture. While in the prevalence of 1956, this tendency could not be demonstrated, the epidemie occurred simultaneously. The two strains “Okayama 56 E” “Okayama 56 F” were isolated from the brain of the patients respectively, and this both strains were serologically identified with “Japanease B Encephalitis Nakayama strain”.

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STUDIES ON JAPANESE B ENCEPHALITIS VIRUS (1)

PRESERVATION OF JAPANESE B ENCEPHALITIS VIRUS BY LYOPHYLIZATION

Hisanori Okuda

1956 Volume 68 Issue 11supplement Pages 7-11
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_7

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1) The Japanease B Encephalitis virus prepared from the brain of the infected mouse was liophilized using dryice and alcohol in the vacuum degree 10^-3mmHg, thereafter kept in 4°-8°C. In such condition, the virus maintain its activity for eighteen to thirty months.
2) If the material kept in glycerol-saline solution for some days before lyophylization, the virus could similarly preserved its activity.
3) The five or twenty percent solution of dry skin-milk was suitable to the medium for lyophylization.

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STUDIES ON JAPANESE B ENCEPHALITIS VIRUS (2)

ON THE SEROLOGICAL REACTIONS BY THE LYOPHYLIZED VIRUS STRAINS AND THE HEMAGGLUTINATION REACTION STORED CHICK ERYTHROCYTES

Hisanori Okuda

1956 Volume 68 Issue 11supplement Pages 13-18
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_13

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1) Making a comparative study of the virus strains which were preserved by lyophilization and the strains serially passaged in the mice brains of several ten generation, the following results were observed.
The formcr strains completely retook their toxity passaging in the mice brains of serial 23 generations. Any difference could not be demonstrated between the both strains in the point of the infective titer to mice. Inoculating the both strains to mice respectively, the titer of the antigen for complement fixation reaction showed the approximately same value, while in the titer of hemagglutinin lyophilized strains showed higher value.
2) For the purpose of preserving the reactivity of chick erythrocytes in the hemagglutination reaction, the following medical solution mixed with chick blood. In case of 2% Na-oxalate, the reactivity was preserved best, 2.5% Na-citrate and 0.1% heparin next, and modified Alsever's solution worst. And yet in the last case, the reactivity was demonstrated after two or three weeks.

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STUDIES ON JAPANESE B ENCEPHALITIS VIRUS (3)

ON THE HEMAGGLUTINATION BY THE SENSITIZED ERYTHROCYTES WITH CHICK SERUM

Hisanori Okuda

1956 Volume 68 Issue 11supplement Pages 19-27
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_19

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1) Adding flesch chick serum to the chick erythrocytes which had lost their reactivity on account of preserving over six weeks in modified Alsever's solution, their reactivity for hemagglutination could be observed again.
2) Even the guinea pig erythrocytes sensitized with chick serum were agglutinated by Japanese B Encephalitis virus, and the hemagglutination was specifically inhibited by the antiserum.

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CLINICAL AND EXPERIMENTAL STUDIES ON JAPANESE B ENCEPHALITIS

2. CLINICAL APPLICATION OF THE NEW THREE DIAGNOSTIC METHODS TO JAPANESE B ENCEPHALITIS PATIENTS (HEMAGGLUTINATION INHIBITION TEST, SALICYLALDEHY D REACTION AND KOMAGOME REACTION)

Atsumaro KIYAMA

1956 Volume 68 Issue 11supplement Pages 29-44
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_29

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Recently the hemagglutination inhibition test with Japanese B encephalitis virus was discovered by Sabin; salicylaldehyde reaction and Komagome reaction in liquor cerebrospinalis from Japanese B encephalitis patients were reported. Thus the progress in diagnosis of Jap. B encephalitis is quite remarkable. Applying these three tests to the patients at Hiraki Department of Internal Medicine, I studied the specificality and clinical signjficance of these tests, and the following results were obtained:
1. Hemagglutination inhibition reactions in serum and liquor cerebrospinalis from Jap. B encephalitis patients become positive about three or four days after the onset of illness; and since then the percentage of positive instances and the inhibitory titers rise day by day, and this test is definite for the diagnosis of Jap. B encephalitis. The hemagglutination inhibition reactions begin to appear as early as in latter half of the first week, in which stage the complement fixation test, that is considered to be definite for the diagnosis of Jap. B encephalitis, remains still negative, and are positve even in the cases in which the complement fixation reactions are not yet positive after the first week, accordingly this hemagglutination inhibition test is more sensitive and useful than complement fixation test.
2. Salicylaldehyde reactions in liquor cerebropinalis from Jap. B encephalitis patients are extremely strong and show a high positive percentage during the first ten illness-days, but since then the positive percentage decreases rapidly. This reaction is not specific to Jap. B encephalitis patients, but this can be explained as a result of particular properties of. Jap. B encephalitis virus, which impairs glucose utilization in the organism. Therefore it can be concluded that this reaction is one of the most important supplementary diagnostic methods in the early stage of illness.
3. Komagome reactions, within the first ten illness-days, result, in not a few cases, in encephalitis type in liquor cerebrospinalis from Jap. B encephalitis patients, but the cases of encephalitis type decrease rapidly since then, and the cases of normal type increase. No definite tendency can be observed in the type change. Its basic causes are as yet uncertain, and further investigation will be necessary to define its clinical significance.

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CLINICAL AND EXPERIMENTAL STUDIES ON JAPANESE B ENCEPHALITIS

3. CHANGES OF OXYGEN CONSUMPTION AND AEROBIC, ANAEROBIC GLYCOLYSIS IN MOUSE LIVER TISSUES, INOCULATED WITH JAPANESE B ENCEPHALITIS VIRUS

ATSUMARO KIYAMA

1956 Volume 68 Issue 11supplement Pages 45-58
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_45

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It is widely known that the Japanese B encephalitis virus invades not only the central nervous system, but also various visceral organs. Among them the liver is the greatest organ, and the metabolism in liver is especially important to organism. Therefore I studied the oxygen consumption and aerobic, anaerobic glycolysis of liver tissues from mice inoculated with Jap. B encephalitis virus by means of Warburg's manometric method; and the following results were obtained:
1. The rates of aerobic, anaerobic glycolysis of liver tissues from mice inoculated with Jap. B encephalitis virus decrease remarkably after seventy two hours after virus inoculation, but the respiration is not impaired at this stage.
2. The Jap. B encephalitis virus impairs glucose utilization of the tissues of the organism, in which the virus invaded, and this impairment causes general ketosis, which is considered to be a cause of salicylaldehyde reaction.
3. The Jap. B encephalitis virus has no iufluence on Pasteur's ferment in mouse liver.
4. Types of mouse liver metabolism, which are classified by Meyerhof's quotient, Warburg's quotient and fermentation excess, are not altered even after the Jap. B encephalitis virus infection.

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EXPERIMENTAL STUDIES ON JAPANESE B ENCEPHALITIS WITH THE RADIOACTIVE P³²

1. INFLUENCE OF P³² UPON THE NUCLEIC ACID QUANTITIES IN THE ORGANS OF MICE INFECTED INTRAPERITONEALLY WITH JAPANESE B ENCEPHALITIS VIRUS, WITH SPECIAL REFERENCE TO THE BRAIN, LIVER

MITSUO MATSUHISA

1956 Volume 68 Issue 11supplement Pages 59-68
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_59

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In this experiment were used the Mice injected intraperitoneally with 25μc of P³² twenty four hours after they were inoculated with Japanese B Encephalitis Virus.
And periodically they were quantitatively investigated. Seven days and ten days after the infection, certain quantities were collected out of each of these organs; and they were fractioned by Schmidt and Thannhauser's method into DNA-P and RNA-P; whose quantity was calculated by Fiske- Subbarow's method.
DNA-P and RNA-P in each organ of healthy mice were used as control objects for this experiment.
When healthy mice were injected intraperitoneally with the same amount of P³², RNA-P decreased only in liver; but, DNA-P remarkably decreased in each organ.
The result due to administration with 25μc of P³² to the mice which were inoculated intraperitoneally with Japanese B Encephalitis Virus previously, was similiar to that of healthy mice
RNA-P decreased a little only in liver, and DNA-P decreased remarkably in each organ. It was also found out that the time of death and of onset were both postponed.
It is widely known that DNA-P increased in each organ when mice are infected with the Virus. But in my investigation DNA-P decreased. So I cannot help concluding that β ray of P³² prevented the multiplication of Japanese B Encephalitis Virus.

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EXPERIMENTAL STUDIES ON JAPANESE B ENCEPHALITIS WITH THE RADIOACTIVE P³²

2. INVESTIGATION OF NUCLEIC ACID METABOLISM BY STHMIDT AND THANNHAUSER'S METHOD IN THE ORGANS, BRAIN AND LIVER, OF MICE INOCULATED INTRAPERITONEALLY WITH JAPANESE B ENCEPHALITIS VIRUS

MITSUO MATSUHISA

1956 Volume 68 Issue 11supplement Pages 69-78
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_69

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Nucleic acid metabolism in brain and liver of mice inoculated intraperitoneally with Japanese B Encephalitis Virus was investigated by Schmidt and Thannhauser's method. P³² was used chiefly as the tracer throughout this investigation.
P³² was intraperitoneally injected 66 and 138 hours after the mice were inoculated with the virus.
After 6 hours, certain quantities of brain and liver were collected to be fractionated by Schmidt and Thannhauser's method into DNA- and RNA-fractions.
The P³² activity and quantities of these nucleic acid fractions were investigated in this experiment.
As the control objects, the healthy mice injected intraperitoneallywith P³² were used.
In the incubation period, an increase of the P³² activity in liver was recognized both in the DNA- and the RNA-fractions, especially in the latter.
This fact shows that, the nucleic acid metabolism of the liver suffers a marked change in the incubation period.
On the other hand, at the acme stage of infection, an increase of the P³² activity was observed in the DNA fraction of the brain. And then, quantitatively, during the incubation period, a marked change was not seen in the DNA-P and the RNA-P, but at the acme stage of infection, both the DNA-P and the RNA-P increased; while no increase was observed in the fraction of control objects.
It is quite interesting that the results of this investigation was the same as those of investigation made by the mice inoculated intracerebrally with Japanese B Encephalitis Virus.

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ON THE INCIDENCE OF JAPANES ENCEPHALITIS AND THE MOSQUITOES COLLECTED BY LIGHT TRAP AT OKAYAMA

SATYU YAMAGUTI, SEIITI INATOMI, MITIYA KIMURA

1956 Volume 68 Issue 11supplement Pages 79-82
Published: November 30, 1956
Released on J-STAGE: March 30, 2009

DOIhttps://doi.org/10.4044/joma1947.68.11supplement_79

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Observations made on the mosquitoes collected by light trap in the Okayama University Medical School campus during a period of six years (1950-55) showed that the endemic of Japanese encephalitis usually breaks out in this area about two weeks to one month after the peak population of Culex tritaeniorhynchus in August, though a few sporadic cases have been reported from within Okayama Prefecture in May through June and September through October. This unusual occurrence of encephalitis suggests that some other vectors like Culex pipiens may take place of Culex tritaeniorhynchus. On an epidemiological basis Culex pipiens has been suggested as a vector. But an attempt to isolate the virus either from Culex tritaeniorhynchus or from Culex pipiens by intracranial inoculation of filtered emulsion of all the mospuitoes collected during the past two years (1954-55) has failed.

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