The methods include; 1)DEAE-cellulose column chromatography at pH 6.2, 2)Isoelectric precipitation at low ionic strength, 3)Elimination of C5 site generation inhibitor and C7 by repetition of DEAE- and CM-cellulose column chromatography, and 4)Iso-electrofocusing. C3 preparation purified by either of these methods was found to contain no detectable contamination of their complement components including C7 and a recently discovered complement inhibitor, C5 site generation inhibitor; both of which are frequently contained in C3 preparation purified by conventional preparation methods. Furthermore, immunochemical analyses such as radioimmunoelectro-phoresis confirmed that these purified C3 contained no serum proteins other than complement components, either.

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For the purpose of investigation of localizing mechanism in allergic reaction, experiments of sensitization with Streptococcus hemolyticus, mixed with anti-tissue serum, were performed. Experiments of long tern sensitization with Strept. hemolyticus were also performed. The results obtained are as follows: 1) Animals were previously sensitized with Strept. hemolyticus and divided into three groups. The first group received a provocating injection of small quantities of anti-kidney (nephrotoxic) serum, antiskin serum or anti-tendon serum. The second group was provocated Strept. hemolyticus alone and the thind group received the combined antigens of Strept. hemolyticus and the anti-tissue sera. Although in three groups some renal changes were found. The changes were most pronounced in the combined antigen group (the third group). However, definite rheumatic changes could not be found anywhere. 2) The long tern sensitization of dog for 900 days with the cultures of Strept. hemolyticus was performed. Dogs showed proteinuria, microscopic hematuria and azotemia. Pronounced histologic changes were found in the kidney: Capillary tufts of many glomeruli were adhered with the capsules, exsudation, cell-prolife ration or cressent formation were also found in some glomeruli. Bowman's capsules showed fibrous proliferation. Some glomeruli were extensively degenerated and remained as hyaline or fibrous masses. Interstitial fibrosis were found in some area. Summarizing the results, it may be concluded that typical clinical or histological changes corresponding to subacute or chronic glomerulonephritis could be produced by long term sensitization with streptcoccal antigens.

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Besed upon the Hamamoto's theory of "Intravascular Antigen-Antibody reaction", were investigated the histological changes in the liver caused by the active anaphylaxis of Rabbit-fibrinogen. On the experimental method, rabbits were sensitized with the homologous rabbit fibrinogen intra-venously in 7 times and then 7 days after sensitizing injection, the provocative injection was made into the portal vein or the marginal ear vein of the rabbits. Rabbits were sacrificed by the total bleeding in 7 days after the provocative injection, and the histological changes of the liver were observed. The risults obtained were as follows: 1) various degree of degeneration of hepatic cells, 2)parenchymal necrotic foci, 3)cell infiltration in the sinusoids, 4)dilatation of central vein and sinusoids. No significant changes were found in the control animals. Changes of clinical examination of present experiment were as follows: 1)decrease of red cells, white cells and platelets, 2)shortening of blood coagulation time, 3)increase of serum fibrinogen. The significant correlation between these patho-histological changes of the liver were discussed.

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In the present report, were investigated the histological changes in the liver caused by the active immunization of auto-fibrin extracts. On the experimental method, immunization in rabbits were performed with auto-fibrin suspension which were obtained from thir own blood, each animals were received ten intraperitoneal injection, these rebbits were sacrificed and the histological changes of the liver were observed. The results obtained were as follows: 1)Various degree of degeneration of hepatic cells, 2)paren-cymal necrotic foci, 3)cell infiltration in the sinusoids, 4)dilatation of central vein and sinusoids, No significant changes were found in the control animals. Changes of the clinical examination of present experiment were as follows: 1)increase of red cells, 2)Prolongation of blood coagulation time. The significant correlation between these patho-histological changes of the liver and liver function were discussed.

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Air-born pollen survey was carried out in Sagamihara city during 1965-1968, according to the standards of the American Academy of Allergy for air-born allergen surveys in USA. 1) Japanese cedar (Cryptomeria japonica), white cedars (Chamaecyparis spp), and pines (Pinus spp), occupied a majority of all pollens. 2) Predominant tree pollens in angiosperms were oaks (Quercus spp), (Zercova serrata), hornbeams (Calpinus spp), alder (Alnus spp), and hackberry (Celits sinensis). Other tree pollens were a few willows (Salix spp), maples (Acer spp), sycamores (platanus spp), chestnut (Castanea spp), chinquapins (Castanopsis spp), walnut (Juglans), cherry (plunus spp), Euptelea and so on. 3) Pollens of grass family were observed abundantly in 2 seasons of may-june and september-october. In the latter season, grass pollens were more abundant than those in the former season. But in the former season, grass pollens were rather going to increase year after year. 4) Abundant weed pollens were sagebrush (Artemisia spp.) short and giant ragweed (Ambrosia elatior and trifida), japanese hop (Humulus japonicum), and freabane (Erigeron spp.) plumpoppy (Macleaya cordata), and common plantain (Plantain asiatica) were also observed frequently. 5) Pollens of Japanese cedar, pine and oak showeda little change on each count every year. On the other hand, pollens of white cedar and japanese elm changed remarkably from year to year. 6) Pollinating periods of pine, oak and japanese elm in spring showed remarkable change according to the atmospheric temperature. But sagebrush in autumn showed little change every year in pollinating period.

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Incidence of sensitization with air born oak pollens was studied at Sagamihara National Hospital. 1) Air born oak pollens were abundant secondary to pollens from Japanese ceder, white ceder and pine in Sagamihara city and there were little difference of those pollen counts according to the year. Oak pollens were from mainly oak KONARA (Quercus serrata), oak KUNUGI (Q. acutissima) of deciduous type (subgenus Lepidbalanus) and oak ARAKASHI (Q. glauca) and oak SHIRAKASHI (Q. myrsinaefolia) of evergreen type (subgenus Cyclobalanopsis). 2) Among 201 patients with allergic disease, the rate of positive intradermal reactions for oak pollen extracts showed 10.4% to oak KONARA, 9.5% to oak KUNUGI and 12.4% to oak ARAKASHI during four months after oak pollinating period. The correlation between each two among these three reactions to oak pollen extracts were very high. (correlation coefficient 0.9) 3) Four cases which revealed positive intradermal and mucus membrane reactions and passive transfer reactions to oak pollen and two suspicious cases which revealed negative passive transfer reactions and positive intradermal and mucus membrane reactions to them were reported. One of the four cases reacted to oak ARAKASHI only, the other cases reacted to 3 oak species. it was presumed from the results of neutralization tests in vivo that they recognized these three pollens as the same antigen. 4) Asthma attacks in five cases of hay asthma due to oak pollen occurred in its pollinating period (April-June) and repeated till July or September in some cases. Two cases had the attacks again in October to December and they also reacted to house dust extract. 5) Four patients with oak hay asthma reacted with till 10^3 to 10^7 time dilution of oak pollen extracts by intradermal tests and PK titers were 1 to 125. These thresholds were lower than the thresholds in patients with weed (sage, ragweed, Japaneses hop, etc) pollinsis. 6) The rate of occurrence that four cases of oak pollinosis were found out among 201 patients with allergic diseases was estimated to be not so low because these kinds of extracts have not been widely used in Japan. Oak pollen sensitive patients might be detected further more following interest and attention in this disease and practical application of these pollen extracts.

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