One of the authors (Akira) and his coworkers¹⁾ performed a study on the structure of a crystalline decomposition product of starch by bacteria. The present authors took up the study on the structure of polysaccharide as one component of bacterial cells of various microorganisms and also its relation to immunogen. The present experiments were conducted in order to study the structure of a polysaccharide obtained as a bacterial component of Bacillus pyocyaneus. In the study of the Flexner group of Bacillus dysenteriae, Walther, Goebel, Binkley and Perlmann²⁾ extracted one part of the dried bacterial cell with ten parts of 50% pyridine and put the extract to dialysis and electrodialysis. By the addition of acetone to its dialyzate to make a 66% solution, they obtained polysaccharides. Studies were conducted on these polysaccharides with amylase and amylopectin by Potter and Hassid³⁾, and by the sodium metaperiodate oxidation of the terminal methyl of glycogen by Akiya and his coworkers⁴⁾ to find out the number of the monosaccharides constituting polysaccharide and the type of their bonding, i.e. whether they are in a long-chain, ring form, net form, or in a dehydrated form. During the structural study on gums, Smith⁵⁾ used a mixture of pyridine and formamide as a reaction promoter and solvent, especially in acetylation. The present authors used the Tsuchijima strain of Bacillus pyocyaneus which was inoculated on an agar slant and incubated for two days at 37°C. The bacterial cells were then collected, treated with ether and alcohol and the residue thereby obtained was treated by the above method and a small amount of polysaccharides were obtained by Goebel's method. The substance thus obtained gave negative iodine reaction and was found to be in a chain form as a result of metaperiodate oxidation. The hydrolysate of this substance obtained with concentrated hydrochloric acid gave negative phloroglucinol and oricinol reactions, and positive orcinol-ferric chloride, indole, Pinoff and Molisch reacttons from which it is surmised that the polysaccharide is a single chain of hexasaccharide.
By examination with paper chromatography, the hydrolysate corresponds to glucose and fructose, and gives α-methylphenylhydrazone of m.p. 152°, α-benzylphenylhydrazone, m.p. 164°, and phenylosazone, m.p. 205°. These three compounds do not show any depression of the melting point on admixture with d-fructose-α-methylphenylhydrazone, d-glucose-α-benzylphenylhydrazone, and d-glucose-phenylosazone, respectively. Since the acetate of this polysaccharide gave mp. 180° with molecular weight of ca. 1200, it is assumed that glucose and fructose are bonded. The position at which monosaccharides are bonded to each other, and whether the monosaccharides and α-or-β-type will be dealt with in the future reports.

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This cercaria was observed by several Japanese workers referring to the study on lung fluke. Cercaria “hei” from Semisulcospira libertina in Gifu Prefecture described by Ando (1951) was the first report. Nakagawa (1915) described this as Cercaria XII from the same snail host in Shinchiku Province, Formosa. He had, at first, considered it as a cercaria of lung fluke, but later on correction was made by himself. Cercaria C described by Kobayashi (1917) from Korea and several areas of Japan and Cercaria C described by Yoshida (1917) from several areas of Japan are also identified with this species. Later, in his review of Japanese cercariae, Kobayashi (1922) characterized this cercaria as Xiphidiocercaria A, while Faust (1924) named it Cercaria nipponensis. Later, Yokogawa and Wakeshima (1934) described it from Formosa as Cercaria VI, Fukui and Shimizu (1936) reported it from Japan as Cercaria V. However, the description of this species by respective authors has yet been very incomplete. The writer recently obtained a large number of this cercaria from several areas in Japan which enabled him to examine its morphology in detail. The following is the redescription of the morphology of this cercaria.

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Numerous papers have been published on the quantitative estimation of amino acid by paper chromatography. One of them is the extracting method by Naftalin and Awapara et al., in which the colored spot is cut off from the paper, the amino acid on the spot is extracted with suitable solvent and the concentration is determined by usual colorimetric method. The other one is transmission method by Fosdick and Bull et al., in which the color density of the spot on the paper is directly measured by electronic transmission densitometer. The method by Fisher is based on a linear relation between the area of the colored spot and the logarithm of the concentration of samples. This method is considered to be the most convenient one. The measurable amount of amino-N by Fisher's method lies within the range of 0.3-3.0γ, but Fisher did not describe it in detail.
We attempted to estimate primarily the amount of aspartic acid and glutamic acid by Fisher's method, but without satisfactory results. After a good number of trials, we found that the reason of this failure was due to the larger area of colored spot than expected. Therefore, we took care of restricting the area of colored spot as small as possible. There are two ways to be considered to fulfill the requirement stated above, that is, the smallest possible amount of the drop on the original point and the selection of appropriate solvent of small Rf value.
We adopted a 1 μl pipette instead of 5 μl and n-butanol as the solvent instead of phenol for certain amino acids, (glycine and alanine which have larger Rf value in the case of phenol) .
Fisher employed a planimeter to measure the area of colored spot, but we used section paper method. The value by our method coincided well with that by planimeter method.

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Ultracentrifugation especially fractional ultracentrifugation is considered to be an excellent tool for purification and concentration of influenza virus, but does not always give satisfactory results, while absorption-and-elution methods with calcium phosphate or with red blood cells comprises some drawbacks beside many remarkable features. It is beyond discussion that the alcohol or methanol precipitation method plays an important role together with the above mentioned methods for the purification and the concentration of influenza virus because of its superiorities on certain view points.
There has been no work worth mentioning about the methanol-preci-pitation of influenza virus since Cox et al applied this method for the purification of this virus for the first time in 1947. But, later in 1950, a paper was again published by them about it, in which they stated that the method proposed previously by themselves had not always furnished goods results. Further experiments were performed extensively and some parts of their previous statement were corrected in the above mentioned paper according to their results. They also stated there that it is the ionic strength of an eluting medium that plays an important role, the study of which had been neglected previously.
Several doubtful points were found in this method during the study of the purification of influenza virus in our laboratory. The present publication is mainly the results of the experiments undertaken to elucidate these points and to qualify several factors for the purification of influenza virus by means of methanol precipitation.

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