The significance of free amino acids within the cell has been clarified by Gale. While free amino acids, e.g. glutamic acid, lysine and etc. could be found quite easily within the cell of Gram-positive bacteria, such free amino acids could not be found within the cell of Gramnegative bacteria. It has been made clear that the amount of free glutamic acid increases within the cell of staphylococci in accordance with the grade of resistance against sulfathiazole.
We found previously glutamic acid as an essential metabolite of dysentery bacilli. It was anticipated that the amount of free amino acids within the cell of sulfathiazole resistant dysentery bacilli would be varied from that of normal cells in view of the results obtained by Gale. The experiments reported in this paper were designed to determine the free amino acids either from sulfathiazole sensitive or resistant strains. Above all, we examined the variation in the amount of free amino acids within the cell of sulfathiazole resistant strain cultivated in the medium containing sulf athiazole.
Since the amount of amino acid to be tested in this report is less than that tested by Gale, who determined the minute amount of glutamic acid by means of glutamic acid decarboxylase, we estimated it by paper chromatography. The method for quantitative analysis we employed was certain modification of the method by Fisher et al. Report on this modification is now being planned for publication sometime in future.

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Now it is well known that viruses which cause paralytic poliomyelitis are not immunologically unique but are divided into at least three immunologically different types, Brunhilde, Lansing and Leon (Bodian) . Recent investigations on non-paralytic polio revealed that polio viruses in a broader sense may include Coxsackie viruses, Col-SK group viruses and some other groups of viruses in addition to the above mentioned 3 types of virus.
Among these 3 paralytic polio viruses, the existence of Lansing type polio virus in Japan was confirmed by positive neutralization test of human sera, collected in various parts of Japan, against the polio virus Lansing strain, and by the isolation of a new Lansing type polio virus “ B 34” strain in Tokyo, which was identified with Lansing strain. The existence of the other types of polio viruses in Japan will be reported in a separate paper.
As for the specificity of the neutralizing substances against polio virus contained in the sera tested, it may reasonably be assumed that most of them were produced specifically after invasion of the virus; this problem will be discussed later in this report. If the above assumption is correct, detection of neutralizing antibodies in animal sera will reasonably serve to support the presumption of the existence of polio virus in Japan, and also to estimate the epidemiological significance of these animals in human poliomyelitis.
The present study was planned to give some explanations on these problems, and also to discuss the specificity of neutralizing antibodies.

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The authors described the structure of a polysaccharide obtained as a bacterial component of Bacillus pyocyaneus in the previous report. The present studies were undertaken for the same purpose with Haemophilus pertussis, which was incubated in Verway liquid medium for three days at 37°C. After examination of grown bacteria, the liquid was centrifuged to remove active charcoal in the medium and bacterial cells collected by de Laval's centrifuge. Fractional extraction of these cells by Goebel's method yielded a g lucoprotein at a concentration of 66% acetone. This glucoprotein was hydrolysed by Morgan's method's by which polysaccharides were obtained. This substance contained no nitrogen or protein, gave negative iodine reaction, and was assumed to be branched by the result of metaperiodate oxidation and the molecular weight of the acetate. The hydrolysate of this polysaccharide obtained with concentrated hydrochloric acid gave negative phloroglucinol, naphthoresorcinol and orcinol reactions but positive Molisch, Dial, indol and Pinoff reactions which allows the assumption of hexasaccharides bonded in a branched chain. By the examination with paper chromatography, the hydrolysate of this polysaccharide corresponds to mannose, galactose and fructose, and gives phenylhydrazone of m.p. 194°, as phenylosazone of m.p. 200°, a-benzylphenylhydrazone of m.p. 156° and a-metylphenylhydrazone of m.p. 145°. The first three showed no depression of the melting points when fused respectively with d-mannose-phenylhydrazone, and d-galactose-a-benzylphenylhydrazone. The last named gave a depression of the melting point upon admixture with d-fructose-a-methylphenylhydrazone. Detailed studies on this point will be made later due to the minute amount of material available. The amount of mannose and galactose seems to be about equal but the amount of fructoselike substance is very small. Time acetate of the polysaccharide showed m.p. 97° and a molecular weight of ca. 3000 from which it is surmised that mannose and galactose are bonded. Detailed studies on the bonding positions of each monosaccharide and their types will be reported in the future.

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Antibiotic studies originated from the observation of antagonistic relation of microorganisms. However, according to Gottlieb and others, the role of antibiotics in antagonism in nature is rather doubtful. There is almost no observation on antagonism between rickettsia and streptomyces, yet the products of the latter, chloramphenicol, aureomycin and terramycin, inhibit the multiplication of the former. Existence of many antibiotics and vitamins in the culture of microorganisms suggests that microorganisms produce various substances which influence a certain course of metabolites processes and inhibit or promote the growth of microorganisms. Among path.ogenes, virus is the particular one in the point of the modus of the multiplication. It multiplies only in the living cell, therefore, its multiplication seems to require some energy-rich metabolic products which can not be taken out of the cell, or to depend upon certain metabolic reactions of the living cell. Though the modus of the multiplication is a particular one as a whole, each of the separate processes may not be particular but it may resemble a certain course of the metabolic processes found in rickettsia and other microorganisms. Microorganisms, which have been found to produce substances inhibiting the growth of other microorganisms or rickettsia, may produce substances which influence a certain chemical process necessary for the virus multiplication.
Several literatures concerning antiviral effects of B. subtilis, A. aerogenes and yeast are cited in the book written by waksman. Horsfall and others found that a polysaccharide of Friedlander bacillus causes inhibition of multiplication of pneumonia virus of mice and mumps virus, but not influenza viruses and Newcastle disease virus. The antiviral effects of bacterial and other polysaccharides are summarized by Horsf all and Ginsberg. It is interesting that their effects are specific to a certain group of viruses. The specific relation of certain substances to viruses is also reported by Kochi and others, and Groupé and other. An antibiotic, viscosin, produced by Pseudoinonas viscosa, which inhibits the growth of acid fast bacteria, inhibits markedly the virus of infectious bronchitis of chickens, and weakly the virus of murine influenza. Ehrlicin, a product of an organism resembling Streptomyces lavendulae, inhibits viruses of influenza and infectious bronchitis but not vaccinia virus. These studies indicate that a certain product of microorganisms is inhibitorily effective to a certain group of viruses.
Among fungi, yeasts, streptomyces and bacteria, Streptomyces is the most fruitful source of antibiotics, particularly of antirickettsial substances. Rickettsia is the pathogene most closely related to virus. Therefore, the writers examined the culture filtrate of streptomyces strains for the antiviral effect both in vitro and in vivo and, as the result, thirteen strains producing a substance effective to western equine encephalomyelitis virus were discovered. Among these thirteen strains, twelve were found to be one same new species and another one was found to belong to Streptomyces rubescens. However, the antiviral substances obtained from these two species were extracted by the same process, appeared in the same fraction of the chromatography, showed the same color reactions, and consequently they were recognized as one same substance. We named this antiviral substance “Abikoviromycin”. When this substance is mixed with the virus and injected to mice intracerebrally, it inhibits the infection at the dilution of 1 in approximately 8, 000, 000. It is effective to viruses of western equine encephalomyelitis and eastern equine encephalomyelitis, but not those of Venezuela equine encephalomyelitis and Japanese B encephalitis. In this paper, characters of the strains and the extraction and nature of the antiviral substance are described.

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