1. As reported previously, histopathological changes of adrenal glands in Wistar Rats (male, 100-200g) were observed with the injection of purified histamine-sensitizing-factor (HSF): Zona glomerulosa, the outer layer of zona fasciculata, and their lipid content were constant. Marked cytolysis in zona glomerulosa. Extraordinary depletion of lipids in the inner layer of zona fasciculata. Zig-zag formetion from the beneath the capsule reaching the deeper region of the inner layer of zona fasciculata (Fig. 1 and 2).
It is well-known that diphtherial exotoxin induces marked hemorrhages of adrenal gland. In refference to this, histamine-sensitizing-factor (an exotoxin) of Bordetella pertussis might cause any change in the adrenal gland. Under this hypothesis, this experiment was carried out. Consequently, the changes mentioned above (Fig. 1 and 2) were recognized. These changes imply disturbance of secretion of hormones (Cortisol, Aldosteron, Adrenal Androgens). I drew inferences from these facts that these were reported as the results of “stress”. I got a lot of letters from everywhere in the world. Unexpectely, among them, the comment of Prof. M.S. Zakharova at Moscow, USSR, was valuable. She critisized that my findings are not only results of stress, but also they are functional changes of the whole host body. It is of the very importance for establishment of prevention against whooping cough infection. This suggestive comment gave me a hint.
No doubt, the changes (Fig. 1 and 2) express only static stage, followed by rapid repairement, accompanying hyperfunction and production of antitoxin. So that, the forced definition “preventive antigen” is completely useless.Recently, Arai & Munoz could crystallize HA (Fimbrial Hemagglutinin), whick lacks completely preventivity. Also, Arai & Munoz could succeed in crystallization of “pertussigen” (Package of HSF, LPF & thermo-labile lethal antigen). The future of pertussigen deserves the keen interest.
2. Is IC (INTRACEREBRAL CHALLENGE) of mice the one and only as a test?
IC method is spread all over the world like the Bible. Oblivious experiments by Standfast whose work I have learned from Dr. Margeret Pittman of NIH, Md, NSA. Standfast's experiments (1) Usual pertussis vaccine gave good effect on IC mouse protection as well as field trial. (2) Contrary to this, pertussis vaccine heated at 100C for one hour do not any effect on IC mouse protection at all. However, this heated vaccine gave good effect on IN (INTRANASAL CHALLENGE) mouse protection as well as field trial (human being). This undeniable facts are of the very importance, because packkage-pertussigen (a group of exotoxins-thermolabile) was vanished away by heating. Hence, one can obtain the most safe pertussis vaccine without any side effect. It reminded of my heat-stable neurotoxin.
I do not know whether it is an exotoxin or not. In spite of unknown character, it shows
charactristic whooping cough syndromes. My working hypothesis suggests that “heat-stable
antigen” by Standfast might coincident with my “heats-table neurotoxin”.

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Moraxella bovisの臨床分離株およびtype strainの溶血物質について検討し,次のような結論を得た。
1. 耳漏由来の臨床分離株20株はウマ,ヒツジ血液に対し100%,ヒト血液に対しては80%,ウサギ血液に対し75%の菌株が溶血性を示した。
2. ハートインヒュジョン培地に酵母エキスを添加することによりM. bovisの増殖および溶血活性は良好となるが,酵母エキスに溶血物質産生促進作用があるとは考えられなかつた。
3. M. bovisの増殖至適pHは7.0∼8.0でありpH 7.5でもつとも増殖が良好であつた。pH 6.5∼7.5の範囲内では溶血物質産生状況に差異は認められなかつた。
4. M. bovisの溶血物質産生能はFe⁺⁺, Mg⁺⁺,還元剤,酸化剤,界面活性剤,レシチン,グルコースなどの影響はほとんど受けなかつたが,トリプシンを菌接種時添加することによつて抑制された。
5. M. bovisのsmooth形集落はrough形集落より溶血物質産生能が大であつた。
6. M. bovisの溶血性は培養後4Cに24時間放置した場合に大であり,とくに30Cで培養した後に低温処理した場合が活性が大であつた。
7. M. bovisの溶血物質は水溶性で菌体内には認められず,耐熱性,ニンヒドリン陽性,塩基性を示す低分子物質である。

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Lactobacillus caseiをStreptomyces globisporus 1829株の培養上清より単離精製されたendo-N-acetylmuramidase (M-1酵素)で処理し生理活性を保持したプロトプラストを短時間にかつ高率に産生する条件を検討した。L. caseiはM-1酵素に対して高い感受性を有し,この感受性はプロトプラストの浸透圧調節物質として使用した蔗糖やラクトースの共存下において著しく亢進した。この観察に基づき,0.9M蔗糖加緩衝液に懸濁したL. caseiをM-1酵素で処理すると,37C, 10分間のインキュベートで処理菌の95%以上がプロトプラスト化した。調製されたプロトプラストの膜構成タンパク質の組成は未処理細胞のそれと変化がなく,また生理活性の指標としたアミノ酸能動輸送能も未処理細胞の輸送能に比べてまつたく差異が認められなかつた。以上の事実から,本実験で調製したプロトプラストは良くその生理活性を保持しており,今後のアミノ酸能動輸送機構の解析や,プロトプラスト融合法を用いた遺伝解析に道を開くものと思われる。

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