Group A streptococci produce extracellular esterase3 (STE), STE-A 1, and STE-A 11, and group B streptococci produce STE-B. A method to estimate quantitatively the antibodies to these STE (ASTE) was devised.
To separate the complex of STE and ASTE from superfluous enzyme and serum components, cell wall debris of staphylococcus aureus, strain Cowan 1, were used as adsorbent. To develop the enzymatic activity of STE bound with ASTE and staphylococcus cell wall, S-acetylthiophenol was used as substrate and 5, 5'-dithiobis (2-nitrobenzoic acid) as coloring reagent.
The present report deals with basic problems of this method and follow-up estimation of ASTE on sera from 88 patients suspected of suffering from scarlet fever. The results were compared with ASO titers estimated on the same specimens.
In 52 of 88 cases ASO titers elevated and in 55 cases levels of ASTE raised significantly. In 48 of 88 cases levels of both antibodies raised similarly. Of 55 cases in which ASTE raised, 43 cases showed the raise of ASTE-AI, and 12 cases the raise of ASTE-AIL
In 32 of 88 cases the levels of ASTE-B raised.

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The total of 94 school children at the first and second grades in Nikko City were followed-up on their carrier state of streptococci and T-antibody during the period of October 1973-December 1977. The sera in 1973, 1974, 1975, and 1977 were tested against T-1, T-3, T-4, T-6, T-12, T-22 and T-28, which were prevalent in the community.
The results were as follows:
1) The sero-positiveness increased in the course of time according to T-type and the year of testing. The positive-conversion rates against T-4 and T-12 were higher than the other types.
2) The total of 6 among 31 positiveness, which were positive-converted at the second year of the observation, were negative-converted after 3 years. However, the T-agglutinin titer of the remained were kept at enough titer, showing the usefulness of T-antibody as a sero-epidemiological tool.
3) The majority of the children, who were already positive at the beginning of the observation, kept the T-antibody quite well after 4 years, especially in case of T-4 and T-12, suggesting the duration of T-antibody positiveness for more than 4 years.

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Virucidal activities of Methanol, Ethanol, Iso-propanol and N-propanol against 6 types of hydrophilic virus and 5 types of lipophilic virus which consisted of 4 types of enveloped virus and one type of Adeno virus were investigated.
Viral inactivation was determined by mixing 0.9 ml of alcohols with O.1 ml of diluted virus. Influenza virus and Herpes simplex virus were diluted with PBS and the other viruses with distilled water. The viral titer of Adeno virus was 4⁶TCID₅₀/0.1 ml and those of other viruses were 10⁶TCID₅₀/0.1ml. After end of test period 9 ml of PBS or distilled water were added to stop further action of alcohols.
Virucidal tests against Influenza virus were carried out by inoculating the virus into the allantoic cavity of the chick embryo and testing for viral hemagglutinins in the allantoic fluids. In case of all other viruses viral inactivation was determined by inhibition of CPE on Hela cells.
The inactivating concentrations of alcohols recorded in the tables practically denote final percentages of alcohols diluted with virus solution.
Lipophilic viruses are readily killed by all kinds of alcohols and the virucidal action increases with the increase in the number of carbon of the alcohols.
It seemed that the sensitivity of lipophilic viruses to alcohols resembled to that of vegetative forms of bacteria. But in this group, Adeno virus which was no envelope, revealed rather strong resistance to ethanol and iso-propanol which required 120 minutes to inactivate it at 20°C.
Hydrophilic viruses are relatively resistant to alcohols and are inactivated only by methanol and ethanol. These two alcohols revealed virucidal action decreased with the increase of the number of carbon of alcohols on the contrary to lipophilic ones. It was recognized, however, AHC virus had less resistancy to alcohols and easily killed by n-propanol. We have observed in the past that the reaction of AHC virus to several germicides was different from other types of Enteroviruses, and in this report too, AHC virus showed different attitude to alcohols.

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The inactivation of viruses by ethanol in organic materials was investigated.
Viral inactivation in liquid materials was determined by mixing 0.9 ml of ethanol with 0.1 ml of Polio virus in undiluted materials. At the end of the test period 9.0 ml of distilled water was added to stop the action of ethanol and to avoid toxicity for cell culture. The percentages of ethanol recorded in the tables indicate final concentrations diluted with the virus material. Inactivation of Polio virus in a clotted rabbit blood and Hela cell sheet were carried out by filling a test tubes containing these materials with ethanol. At the end of sensitization, it was replaced immediately by 2.0 ml of M.M. and was frozen by aceton dry ice. After sonication these materials were centrifuged and the supernatant were used for estimation of surviving viruses. Inactivation of viruses in dry materials was determined by pouring 1.0 ml of ethanol on dry calfserum smear containing known titer of NDV and Vaccinia virus on the bottom of wide mouth bottle. At the end of test period 9.0 ml of distilled water was added for dilution of toxicity and cessation of the action of ethanol.
The virucidal activity of ethanol on viruses in either humoral or solid organic materials increased with the increase of concentration of ethanol, when these materials contained water sufficient for the action of ethanol. By 90% ethanol Poliovirus type 1 in an undiluted calfserum was inactivated so rapidly as in PBS, but 70% ethanol required more than ten times of sensitizing period, compared with in PBS. A similar phenomenon was observed in solid materials. Polio virus in an infected Hela cell sheet having titer of 10^7.8 TCD, reduced its infectivity to 10^1.5 TCD, by exposure to 99.5% ethanol for one minute, whereas by 70% ethanol it remained at the titer of 10^5.5 TCD. Polio virus in blood clot was inactivated almost similarly as in Hela cell sheet.
On the othei hand when viruses were in dry materials rather low concentration (40-60%) of ethanol was more effective than the higher concentrations, though the highest efficiency depended on the susceptibility of the each viruses to ethanol.

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In my previous paper, it was reported that the isolation rates of GBS in the vagina tend to increase by age. This study was done to assess whether it was an important facter for increase of the isolation rate of GBS so that lactobacilli became less predominant and the vaginal PH became higher by age. And the influences of lactobacilli on other flora and the variations of other flora by age were studied clinically and bacteriologically.
The subjects of this study were 218 normal, healthy women who visited the Department of Obstetrics and Gynecology at the Central Hospital of Japanese National Railways between October 1977 and September 1980. By discharge samples obtained from the vaginal lateral wall, aerobe, anaerobe and fungi were cultured and isolated. Vaginal PH was determined by a paper method using BCG and BTB as an indicator.
The distribution of microorganisms in 218 women has been shown in the descending order of frequency as Lactobacillus (53.7%), S. epidermidis (38.7%), E. coli (22.1%) and Corynebacterium (18.9%) were present as heavy growth. GBS was isolated in 6.5% of cultures.
GBS, E. coli, Bacteroides and S. aureus were isolated significantly higher from lactobacillus-negative women than from lactobacillus-positive women.
The isolation rates of lactobacilli tend to increase and vaginal PH tend to become significantly higher by age.
In our findings, age-influences on vaginal microbial flora were evident. Significantly higher rates of lactobacilli were detected among younger women than among older ones. But GBS were detected more frequently in older women than in younger ones.
No significant variations of isolation rates of E. coli and Bacteroides by age were recognized, but S. aureus were isolated frequently among women older than 50 years.
The growth of GBS and S. aureus was suggested to be restricted by lactobacillus. But candida albicans was isolated frequently among younger women and its isolation rates tend to decrease by age. It has been suggested that lactobacillus did not restrict the growth of candida. Lactobacillus and candida would be able to coexist in the vagina.

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