In earlier papers hitherto published on this serial study, the author and the co-workers have reported that the Japanese B encephalitis virus can easily multiply in Ehrlich ascites tumor cells propagating in mice. Furthermore, they reported that the infection of these tumor cells with the virus could be established in a moment, and once the first step of the infection was accomplished, the cell infected virus instantly enters such a state, that it can not easily be neutralized by the hyperimmune serum of enough neutralizing potency against the virus, although the virus existing in a free state can readily be neutralized by the same serum. Such an infected virus can persist in a latent state for considerably a long period of time in the tumor cells that are existing in mice which contain the neutralizing antibody of significant potency.
In the present study the author performed experiments in order to investigate whether the infection of cells with the virus can also be established in a case as such, that the cells had previously been in contact with the neutralizing antibody for a certain period of time, and afterwards encounters with the virus that comes into this environment.
Experimentations are as follows; i.e. 1.0ml. of undiluted hyperimmune rabbit serum against Japanese B encephalitis virus and 1.0ml. of packed mass of washed Ehrlich ascites tumor cells of mouse were mixed and incubated at 37°C for 30min. After the incubation, to each of these mixtures was added each of decimal dilutions of Japanese B encephalitis virus, from 10^-1 to 10^-7 in amount of 1.0ml. respectively. The mixture consisting of the antiserum, the cells and the virus, was again incubated at 37°C for 30min. After that, the mixture was centrifuged. The neutralization of the virus existing in free state in the mixture of the respective virus dilution was testified by the direct intracerebral inoculation of the supernatant of each of them to mice. The cell adsorbed virus in each virus dilution was put to the demonstration test by the indirect in vitro cell virus infection method, described as follows. The cells in the sediment were washed three times and were inoculated each to three normal mice in certain amount intraperitoneally. Four to 7 days thereafter, the tumor fluid of the mice of each group was drawn, pooled and centrifuged respectively. The supernatant was put to the demonstration test of the virus with the direct method of mouse intracerebral inoculation. When the result of this was positive; i.e. if the virus was demonstrated in the ascitic fluid, the cells that had been intraperitoneally inoculated 4-7 days ago, were designated to have been infected with the virus, although the trials of demonstration of the free virus in the same respective mixtures had resulted in negative.
The results are as follows; i.e., the cell adsorbed virus could be recovered by the indirect method from the mixture consisting of the cells, the virus and the immune serum of rather surplus potency, while the free virus could never be recovered from it by the direct method. In a word, it was demonstrated that the Japanese B encephalilis virus could be adsorbed to the Ehrlich ascites tumor cells that had been suspended in immune rabbit serum or immune mouse ascitic fluid of sufficient potency against the virus and had been in contact with the antibody, and could establish the first step of the virus infection of the cells in this environment.
There remains much to be made clear as to the mechanism of this phenomenon, but standing upon the results obtained so far, it may be possible to conclude, that the first step of virus infection of cell can be established in the existence and under the action of the neutralizing antibody against the virus, so far as at least the Japanese B encephalitis virus and the Ehrlich mouse ascites tumor cells are concerned.

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OUT of 23 strains of Pseudomonas aeruginosa, 14 strains were pyocinogenic strains and 15 strains were sensitive strains to pyocin. However, there were no strain both no-pyocinogenic and insensitive to pyocin.
No sensitive strain was found a pyocin produced by bacteria that are sensitive to it.

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1) Quantitative methods of pyocin activity can be used by the colony counts method which was determined from the relative values of survival becterial colonyes after treatment with pyocin, and also can be determined from the values of the turtidities of logalithmic growth of sensitive bacteria after incubation with adequate amonunts of pyocin. Besides the semi-quantitative methods, spot test and serial dilution method, can be used for determination of pyocin activities roughly.
2) The enriched media can be used for pyocin production. That is, “Poly-peptone” external dialysate solution is added to glutamic acid-glucose-salts media. The maximum activity (units) was obtained in this culture fluid.
3) Purified pyocin can be obtained by the methods of ammonium sulphate precipitation, zone-electrophoresis and ultra centrifugation, and this sample shows homologus by means of electrophoresis and ultra centrifuge analysis. It contains 10.5 per cent of nitrogen and is 400, 000unit/mg. of activity, This activity is not destroyed by means of “Nagase” treatment.
4) Anti pyocin serum can be obtained by injection of pyocin to rabbits.
5) Pyocin is stable at the pH 7-8, but its activity is rapidly decreased at pH 2.2 even room temperature. Pyocin activity is not decreased at 55°C for 30min. but is decreased at 60°C.
6) Decrease of pyocin activity during the process of lyophilization can be protected by addition of glucose, sucrose or Koethoff's buffer (pH 7.2).

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1) Purified pyocin was adsorbed onto sensitive bacteria, Pseudomonas aeruginosa P-2 (P-2), and these bacteria were killed by it, when they are mixed at room temperature.
Pyocin was also adsorbed onto sensitive becteria at 0°C but the bacteria were not killed by adsorbed pyocin at this temperature.
2) Both insensitive bacteria, Ps. aeruginosa P-1 (P-1), and pyocinogenic bacteria of this pyocin, Ps. aeruginosa P-15 (P-15), did not adsorbed any pyocin under the adsorbable condition of pyocin onto sensitive bacteria.
3) Specific pyocin inhibitor was found in the autolysate of sensitive bacteria (P-2), but same inhibitor did not found both in the autolysate of insensitive bacteria (P-1) and pyocinogenic bacteria (P-15).
This specific inhibitor did not precipitate at 20, 000G for 60min., but precipitated at 100, 000G for 90min., and this activity was heat stable one.

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While in infection of cells of Salmonella anatum 293 (=A) with ε¹⁵[A], phage ε¹⁵ establishes lysogeny in about 3 hours after infection, in infection of cells A carrying prophage ε³⁴ cells which survive infection continue to segregate phage-carrier and noncarrier cells in their progeny even for several hours after infection. In contrast, however, when cells A(ε³⁴) are infected with ε¹⁵ which are propagated on A(ε³⁴), lysogeny is established in about 3 hours, even at a multiplicity of infection of 5 or less. These suggest a correlation between host-controlled variation and lysogenization.
When cells A are infected at first with ε¹⁵ and then 30 minutes later with ε³⁴, ε¹⁵ establishes lysogeny in about 3 hours without being affected by ε³⁴, indicating that the genetic function of ε³⁴ may not be responsible for the interference with prophage establishment of ε¹⁵[A] in cells A(ε³⁴). On the other hand, in this experiment, ε³⁴ is interfered with rapid establishment of lysogeny. And in contrast, ε³⁴ establishes lysogeny soon after infection when ε¹⁵-infected cells are infected with ε³⁴ 180 minutes after ε¹⁵-infection and/or when cells A(ε¹⁵) are infected with ε³⁴, indicating that the rapid prophage establishment of ε³⁴ is mainly attributed to the genetic properties of the phage itself.
There is no remarkable difference in time required to establish lysogeny between in infection of cells A and of cells A(g₃₄₁) with phage ε¹⁵[A(g₃₄₁)], indicating that prophage g₃₄₁ does not affect ε¹⁵ on its establishment of lysogeny.
Similarly, g₃₄₁ establishes lysogeny rapidly after infection of cells A as well as cells A(ε^y). It suggests that the rapid prophage establishment is attributed to the genetic properties of phage g₃₄₁.
The above findings lead to the conclusions that the process of lysogenization may be affected by the presence of the genome of unrelated phage.
Notes: A(ε³⁴), A(g₃₄₁), A(ε¹⁵), and A(ε^y) indicate strain A lysogenized with ε³⁴, g₃₄₁, ε¹⁵, and ε^y, respectively.
ε¹⁵[A], ε¹⁵[A(ε³⁴)], and ε¹⁵[A(g₃₄₁)] indicate phage ε¹⁵ propagated on strain A, A(ε³⁴), and A(g₃₄₁), respectively.

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For the purpose to prevent the serious influenza infection among newborns, the following experiment to testify the possibility of the materno-fetal immunization against influenza were performed. Pregnant women at their 8-9th month of gestation were injected with commercial trivalent (A1, A2, B) type vaccine and adjuvant added bivalent (A2, B) type vaccine, specially prepared by Chiba Serum Institute for experimental purpose. These vaccine have the potency of 100CCA/ml and 200CCA/ml per each type, respectively. The mother bloods taken just before the injection and at delivery, and cord bloods and colostrum collected at delivery were tested for their contents of influenza CF-V, CF-S and HI antibodies.
As the results, the increase of CF-V and HI titer in the mother bloods were observed after vaccine injection, especially when subcutaneous 1ml injection following two intradermal 0.1ml injection of routine commercial vaccine, or when 0.5ml adjuvant added vaccine were injected subcutaneously. On the contrary, the increase of CF-S titers were not observed.
In the cord bloods somewhat higher titers of CF-V, somewhat lower titers of CF-S and nearly equal titer of HI antibodies were demonstrated, as comparing with those in the mother bloods.
In the mother colostrum, HI antibodies of lowere titers comparing with those of mother bloods were demonstrated.
From above data it became evident, that the materno-fetal immunity against influenza is completely established by active immunization of pregnant women at their 8-9th month of gestation.

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Myc. sp. Jucho in logarithmic phase was infected by phagus choremis, Hauduroy, and was incubated with vigorous aeration for about 24 hours at 37°C. The titer of thee lysate reached usually 1-2×10¹¹ particles per ml. Phage was concentrated by the three times centrifugation at 17, 000rpm for 60 minutes and was carefully purified by the DNase and RNase treatments.
This phage particle has the cocoon shaped head of ca 95×150mμ in size and tail of 180-190mμ in length.
Extraction of DNA was caried out following the Mandell et al's phenol method.
It was revealed that phagus choremis did not conain RNA, and DNA content per particle was calculated as about in the same order as T-even phage.
The base composition of this preparation was determined by the quantitative paper chromatography showing the base ratio of G:A:C:T=33.44:17.21:33.44:15.91. Ratio of (A+T)/(G+C) was quite similar to that of the host cell.
Relation of the base compositions between host cell and phage was discussed.

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