On many experiments I conclude that there is not always close relation between infectivity on chickembryos or mice and CCA potency of influenza viruses. CCA activity, infectivity and immunizing ability have their own indifferent factor and they can be separated.

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When influenza viruses (A type PR8, A-prime type FM1 and Matsumoto, B type Lee) are denaturated by ultraviolet irradiation or urea, their infectivity is easy destroyed but their hemagglutinating activity is relatively reserved. On their immunizing ability we must study more on detail.

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It is the well established fact that after the recovery from the typical course of the Tsutsugamushi disease the patients acquire the solid immunity against it which was proved to last at least for a decade or thereover. However, in regard to the aspect of the immunity development in the case in which the disease was suppressed with antibiotics as soon as the specific symptoms became manifest, there remains much to be made clear.
The author, in order to make this aspect clear, made some trials on human beings. Namely the patients of general paralysis, which had to receive the fever treatment, were infected with the inoculation of the virus of the Tsutsugamushi disease, Pescadores strain, of certain titer of mouse LD 50, and received the drug therapy with antibiotics to recovery at the various stadium of the disease. The investigation of the immunity development just mentioned was performed upon them. A part of the results obtained is presented preliminary in this paper as the first report. Namely it can be described summarily as follows.
1. The experimental minimal infectious dosis (M. I. D) of the virus of the Tsutsugamushi disease to human beings is of approximate value to mouse LD 50 titer of it.
2. Below the M. I. D., the virus cannot elicit the immunity development at all in the human body, althogh it had certainly invaded into it.
3. The specific symptoms of the disease become just manifest when the invaded virus multiplied somewhere in the body to such an extent to elicit the rickettsiemia of approximately 10^-2 LD 50.
4. The patient is far more sensitive to the drug therapy with antibiotics at the height of the disease or still later than at the beginning of it.
5. The specific reaction of the site of the skin, where the virus had certainly been inoculated, can sometimes be failed, though the inoculated person contracted the disease typically.
6. A certain correlation can be observed between the length of duration of the disease and the degree of rise of OXK agglutinin titer.

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I. Brain and spinal cord of mouse infected and paralyzed with poliomyelitis virus Lansing strain were inserted into bananas and were orally administered to 9 cynomolgus monkeys. Tokyo strain virus (type 2) was also administered to 6 monkeys in the same way. None of of all the 15 monkeys manifested the signs of apparent infection by the oral administration of the virus.
II. Viremia was recognized on 6 of 15 monkeys during the period ranging from the very day of the virus administration to 4-5 days after it. Also of 3 among 7 groups the viruses were positive in stool specimens only on the next day of the virus administration.
III. Two monkeys which had inoculated with cow serum γ-globulin were both viremia negative, and their stool specimens were also virus negative.
IV. Noticable increase of neutralizing antibody of the monkey sera was not observed after the oral administration of the viruses.

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A poliomyelitis virus strain was isolated by intracerebral and nasal administration on monkey from the spinal cord of a paralytic poliomyelitis case died in Scptember, 1950 in Tokyo, which was stored in dry ice until the day of inoculation in March, 1952. The spinal cord of the sacrificed monkey was successfully transfered to mouse by intracerebral inoculation.
The mouse adapted Tokyo strain was applied to the neutralization test against each immune monkey serum of Lansing, Brunhide, Leon, B-34 (a type 2 poliomyelitis virus, recovered in Tokyo, 1948, using mouse only.) and also of Tokyo strain. The result of the neutralization test showed that the newly mouse adapted Tokyo strain was well neutralized by the immune sera of Lansing, Tokyo and B-34, and hardly neutralized by Brunhilde and Leon immune sera. Hence it is understood that mouse adapted Tokyo strain is to belong to type 2 together with B-34 and Kawasaki strains.
At the brain passage, the harmonic mean of the incubation period of this Tokyo strain is 5.4 days which shows 2 days shortening, comparing with that of Lansing strain which is 7.3 days.

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Five nitrogen mustards; methyl bis (beta-chloroethyl) amine hydrochloride, tris (beta-chloroethyl) amine hydrochloride, amyl-bis (beta-chloroethyl) amine hydrochloride, diethyl (beta-chloroethyl) amine hydrochloride and N, N'-tetrakis (beta-chloroethyl) ethylen diamine hydrochloride, and N-oxide of each (N, N'-dioxide with the last amine) were examined in vitro concerning VH or virus inactivating effect (The compounds are referred to only by their initial letters. and vaccinia hemagglutinin is referred to as VH.).
Tetrakis was found to be the most effective in inactivating vaccinia hemagglutinin, whereas diethyl. ox was scarcely effective even at the concentration of 1/25 M. VH inactivating activity appeared to depend on the number of betachloroethyl radicals and the N-oxide of each compound was in general less effectiev than the original compound with the exception of tris and its N-oxide. Amyl, which has the same structure as bis except that the former has an amyl-radical in place of methyl-radical in the latter, was far more effective than the latter. A gross parallelism was observed between the VH (inactivating effect) and viral infectivity inactivating effect of the compounds, but diethyl and diethl. ox were effective in inactivating viral infectivity in vitro at the concentration (where VH was hardly inactivated).
The order of the compounds in regard to VH inactivating activity will be described as follows: tetrakis>amyl>tris. ox>tetrakis. ox>amyl. ox>tris>bis>bis. ox>diethyl>diethyl. ox.
The effect of potassium periodide, hydrogen peroxide or Fenton's reagent on VH was also examined and it was proved that VH was inactivated by potassium periodide, whereas hydrogen perioxide or Fenton's reagent was ineffective under the same conditions.

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Vaccinia hemagglutinin, either so-called thermolabile or thermostable, was inactivated by ultraviolet irradiation.
Inactivation was retarded by a certain dialysable substance contained in normal or infected chorioallantoic membrane and was accelerated by sodium chloride in the suspension.
Sodium chloride was also proved to be an important factor which inactivate vaccina hemagglutinin when heated at 56°C for 30 minutes.

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Potency of Japanese B encephalitis vaccine is tested as follows: On 1st and 4th day, intraperitoneal injection with O.1cc of vaccine respectively, into 30 mice of 3 to 4 week-old, and on 8th day, the test animals as well as 30 mice in control are respectively challenged intraperitoneally with 0.2cc of 10^-1 dilution of mouse brain saline suspension infected with Nakayama strain of Japanese B encephalitis virus. On 22nd day, the test animals must survive over 50% under the conditions of virus infectivity in control being that the LD₅₀ is over 10^-3, for example 10^-26, and also the mortality in 10^-1 is over 90%. When the LD₅₀ is under 10^-4, the test must be retested. When the test animals survive over 50%, the vaccine is good.
By using this potency test, authors researched on the antigenic variation of Nakayama strain of Japanese B encephalitis virus.
Mouse brain passaged virus (M: Standard line) was serially passaged through embryonated eggs, and the virus of 2nd, 10th or 20th generation of passage was respectively designated as E₂ E₁₀, or E₂₀.
The vaccine prepared from E₂, E₁₀, or E₂₀ was respectively designated as E₂Vac, E₁₀Vac or E₂₀Vac.
The virus of egg line was serially passaged through mouse brain, and the virus of 2nd., 10th., or 20th, generation of passage was respectively designated as CM₂, CM₁₀, or CM₂₀. The vaccine prepared from CM₂ CM₁₀, or CM₂₀ was respectively designated as CM₂Vac, CM₁₀Vac, or CM₂₀Vac.
The vaccine prepared from M was designated as MVac.
Cross immune tests between E₂Vac, E₁₀Vac, E₂₀ Vac, or MVac and E₂, E₁₀, E₂₀, or M are carried on.
In the case of challenge with M, the potency of vaccines are as follows: E₂ Vac is highest, E₂₀ Vac lowest and E₁₀ Vac intermediace.
That is, the potency decreases parallel with the increase of aggpassage generation. In the case of challenge with E₂, E₁₀, or E₂₀, however, the potencies of E₂ Vac, E₁₀Vac and E₂₀ Vac are almost the same.
Cross immune tests between CM₂ Vac, CM₁₀ Vac or MVac and CM₂, CM₁₀, CM₂₀ or M are carried on.
The potency of MVac is highest in the case of challenge with M or CM₂₀, lowest in the case of challenge with CM₂ and intermediate in the case of challenge with CM₁₀.
CM₂₀ Vac shows the same tendency as MVac.
The potency of CM₂ Vac is highest in the case of challenge with CM₂, and decreases in due order of CM₁₀, CM₂₀ and M.
From the results mentioned above, it is concluded as follows: the antigenicity of mouse brain passaged line of Nakayama strain of Japanese B encephalitis virus varies by serial passage through embryonated eggs and the variation is revresible.
As the antigenic change is caused by host-alternating, it is considerad as “host-controlled variation.”

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In tissue cultures of rabbit, guinea pig or mouse tissues, either adult or embryonic, multiplication of the Newcastle Disease virus has never been evidenced. In these cultures, the end titer of virus after different periods of incubation was always found lower. than the virus titer at the start. In the presence of rabbit brain, however, the end virus titer after a few days incubation was usually a little higher than in the control culture which contained no tissue. This is considered to be due to a favourable influence of the tissue on the survival of the virus in the medium. Such an effect of tissue remained unaltered after the tissue was dipped in the boiling water bath for five minutes.
In the culture with chicken embryo tissue fragments (brain mostly used) simply suspended in Tyrode solution, the virus multiplied easily, and the culture fluid reached the virus titer of 10^-6.5-10^-7.5 (chicken embryo MID) within 3 days of incubation. In this culture, the same level of maximum virus titer was obtained from different amounts of starting virus, i.e. 10², 10⁴ and 10⁶ MID. Addition of embryo extract and/or serum, or fixation of tissue fragments in blood plasma or on a paper piece did not appear to increase the maximum virus titer of the culture. When the tissue previously dipped in the boiling water bath was used, the virus never multiplied, but maintained the same level titer as in the rabbit tissue culture.
In the culture with abult chicken tissue fragments (spleen mostly used) suspended in Tyrode solution, multiplication of the virus was demonstrated by periodical titrations of the culture fluid, though the maximum titer (about 10^-4.5) never reached the level so high as in the culture of the embryo tissue. In this tissue culture, however, multiplication of the virus was remarkably increased, when chicken serum was added to the medium, or the, tissue fragments were fixed on the paper. By the combination of these two conditions, the virus titer became comparable with that which was obtained in the embryo tissue culture.

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1. 膿病蚕血液上清免疫家兎血清は病蚕血液中に含まれるウイルスを中和する. またこの混合液を蒸溜水で稀釈した場合dilution phenomenonはみとめられなかつた.
2. 膿病蚕血液上清免疫家兎血清中の中和抗体は病蚕血液上清によつて吸収されるが正常蚕血液上清では吸収されない.
3. 膿病蚕血液上清を材料としてフオルマリンワクチンを調製すると多少の免疫効果を示すが, この免疫蚕血液上清中に中和抗体は殆んどみとめられない.

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1. 膿病蚕血液上清疫家兎血清, 多角体溶解液免疫家兎血清は共に多角体を凝集し一方の血清の凝集素は他方の抗原によつて吸収される.
2. 正常蚕血液上清免疫家兎血清は多角体を頭集しないしまた正常蚕血液上清免疫家兎血清及び多角体溶解液免疫家兎血清中の凝集素を吸収しない.
3. 病蚕血液上清を材料としてフオルマリンワクチンを調製し蛹を免疫したが多角体凝集素の増加は殆どみられなかつた.
4. 従つてフオルマリンワクチンを蛹に接種する場合多少の免疫効果を示すのは体液の中和抗体や多角体凝集素のみからは説明しえないことについて考察を加えた.

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1. 膿病蚕血液上清免疫家兎血清を蛹に注射したのち種々時間にウイルスで攻撃したが被働免疫は成立しない.
2. ウイルス接種後種々の時間にまた回数をかえて上記免疫血清を注射して血清療法を行つたが成功しなかつた.
3. ウイルスを50℃或は60℃に加熱すると活性は著しく低下しまた蛹は加熱しても膿病の発生はみられないので, ウイルス階段稀釈液を蛹に接種したのち直ちに50℃ 15分処理したが稀釈ウイルス液の場合でも加熱による療法は成功しなかつた.
4. ペニシリン (900u/ml及び9000u/ml). ストレプトマイシン (953mcg/ml) 水溶液とウイルスを等量に混合し室温30分放置したのち蛹に接種したが潜伏期間の延長, 発病蚕数の減少はみとめられなかつた.
5. 病毒血液上清を材料としてフオルマリンワクチンを調製しこれを接種したのちウイルスで攻撃した. この場合ワクチンを接種したとぎからウイルスで攻撃するまでの時間. ワクチン接種回数をかえて検討したがワクチン接種による予防効果をみとめた.
6. 被働免疫, ワクチン実験, 血清療法等の結果をみると昆虫においてウイルス免疫は細菌免疫の場合とかなり趣を異にしているものと思われる.

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純化したTMVに紫外線を照射しそれがこのウイルスの病原性及び抗元性に与える影響について研究した. ウイルスの感染力はNicotiana glutinosaへの半葉法による接種試験で調べ, 抗元性のテストには沈降反応 (混合法) を用いた.
紫外線照射による活性TMVの残存量と照射量との間には指数凾数的な関係が認められた. 即ち一方に活性ウイルス残存量の対数を取り他方に照射線量をとつた半対数グラフに於けるウイルスの紫外線による障害曲線は大体単純な直線となつた. この結果から, 紫外線照射によるTMVの不活性化は個々のウイルス粒子が少くともsingle ultraviolet quantumを吸収すれば起り得ると推定される. TMVの紫外線による障害曲線は照射時のTMV濃度が低い時その傾斜はより急になつた. これらの曲線から得た不活性化線量 (37%線量) は大体0.2%のTMV濃度の際には2.5×10³μW, 0.02%の時は1.5×10³μWであつた.
抗元性は感染力喪失後も同様に認められたが13.0×10⁵μW以上の線量では完全に失われた. 抗元性喪失の寸前に5.2×10⁵μW照射のものを中心にしてその前後に比較的明瞭な抗元性の増強が認められた.

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Lysogenic strains, E (α₁, β) and R (α₂) could be obtained from their original strains, E (β) and R, respectively by lysogenization caused by infection with Phage α liberated from lysogenic strain A (α).
Endotoxins of each strains were prepared in electrophoretically homogeneous states for precipitin tests. The endotoxin of strain E (α₁, β) was found to react with the anti-serum against the endotoxin of strain A (α), which didn't show any reaction with the E (β) cndotoxin; that is, the endotoxin of strain E (β) was considered to be so modified by the lysogenization with phageα as to possess a common serological specificity with that of strain A (α).
The similar relationship was obtained between the A (α) endotoxin and that derived from R (α₂) strain.
Our previous studies have shown that the purified endotoxin of Pseudomonas aeruginosa seemed to be mainly composed of polysaccharidenucleoprotein complex and that desoxyribonucleic acid (DNA) component could be readily split off by peptic digestion.
It was found that the split endotoxin had lost the aquired common serological specificity, suggesting the DNA fraction might be related to the modification of antigenic properties of the endotoxin.

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