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In the serological assay of insulin antibody, the author studied on the detection of optimal conditions of the bis-diazo-benzidine hemagglutination technique by Arquilla & Stavitsky. With this method, the demonstration of insulin antibody in the sera of immunized rabbits and insulin treated patients were performed. The sera of 11 rabbits which immunized with insulin in Freund's adjuvant & alumprecipitated insulin had weak neutralizing activity to exogenous ox insulin, whereas hemagglutination titres were generally low. Eight rabbits were immunized with insulin-conjugated sheep erythrocytes by use of bis-diazo-benzidine. High hemagglutination titres and no ability to neutralize the physiological action of insulin were demonstrated in all antisera. With this result, it was suggested the hemagglutination titres and the neutralizing activity to insulin were presented independently. The sera of 46 insulin treated patients were tested by the hemagglutination technique for insulin antibody. Only 2 cases were found to have the hemagglutination titres of 1/64 and 1/256. There was no correlation between hemagglutination titres and daily requirement of insulin. Atempts to demonstrate precipitin band by agar gel diffusion method in antisera of rabbits and patients were all unsuccessful.

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The antigenicity of insulin in guinea-pig was studied. A group of ginea-pigs was immunized with insulin conpled sheep erythrocytes by bis-diazotized benzidine and an other group was done with insulin in Freund's adjuvant. The antibodies were demonstrated by BDB-hemagglutination test, Schultz-Dale's reaction, agar gel diffusion and two tests for insulin neutalization, in which the neutralization of exogenous ox insulin was demonstrated by mouse convulsion test and the neutralization of endogenous insulin in mouse was measured by hyperglycemia after injection of the antiserum. Postive Schultz-Dale's reactions and negative agar gel diffusion tests were resulted in both antisera. Antiserum immunized with insulin in Freund's adjuvant had low BDB-hemagglutination titer, positive mouse convulsions test and positive neutralizing activity of endogenous insulin in mouse. High correlation was observed between the BDB-hemagglutination and the neutralization titer of exogenous insulin in mouse. On the other hand neutralization titer of exogenous insulin was independant. The antiserum immunized by BDB-method had extremely high BDB-hemagglutination titer, moderate neutralization titer of exogenous insulin in mouse, and no neutralizing activity of endogenous insulin in mouse, no correlation between the results of three methods was found. These discrepancies suggested that BDB conjugated insulin had few biological activity and that guinea pig antisera reacted in a different way two endogenous mouse and exogenous ox insulin.

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The present investigation was carried out for the purpose of presenting information about the relationship between the morphological changes and the results of functional tests of the liver. Based upon the the theory of "Intravascular antigen antibody reaction", anti-rabbit red cell guinea pig serum (hemolytic titer: from 64 to 32) were injected through the hepatic artery or portal vein of rabbits. Both histological changes in the liver and changes in various liver function tests caused by this reversed anaphylaxis were simultaneously investigated at intervals of 7, 15, (Group IV) 30, 40, (Group III) 60, (Group II) and 90 days (Group I) after a single provocative injection of antiserum. In the histological examination of the liver in rabbits, 15 days after the injection, the granulation tissue with giant cells were seen remarkably surrounding the large area of focal necrosis and fresh fibrosis were developed. Various degree of degeneration of liver cells were found. In a few cases the productive endoarteritis and the fibrinoid swelling of arterial walls were discovered in the interlobular connective tissue. Thirty days after, the small necrotic foci and thrombosis were presented. Ninety days after, the major changes were remarkable fibrosis and the liver showed the postnecrotic cirrhosis at least in part. But in other part of the liver lobules localized limited inflammatory processes were remaind, and the cords of parenchymal cells were somewhat dissociated. On the contrary no significant changes were found in the control animals by injecting the normal guinea pig serum. Then the electrophoretic pattern of serum protein fractions were as follows: a) decrease of total serum protein in Group I and Group II, b) declination of albumin in Group I, c) increase of gamma globulin in Group I and Group II. The results of the liver function tests were positive in Group III only by the Cephalin-cholesterol flocculation test. The significant correlation between these patho-histological changes and the grade of functional disturbances of the liver were discussed.

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Intradermal tests with house dust extract (Torii & Co, Ltd.) were carried out in 53 bronchial asthmatic patients and 5 urticaria patients. Positive results were obtained 62.2% in bronchal asthma, 40.0% in urticaria. Of these positive patients, 18 bronchial asthmatic patientis and 2 urticaria patients have been treated with the repeated injection of house dust extract. Two urticaria patients have all cured. Of 18 bronchial asthmatic patients, 12 patients were estimated; no effective in 3 cases, moderately effective in 6 and markedly effective or cured in 3. No side effects were observed in all cases.

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