Polysaccharides were separated from mycelia and culture filtrates of the filamentous fungi Aspergillus niger and Penicillium chrysogenum, and purified by column chromato-graphy on Sephadex G-50, DEAE- and CM-cellulose, successively. No nitrogen and phos-phorus were detected in the polymer, and the sugar components were observed to be galactose and mannose. The polysaccharides were confirmed to be galactomannans which were easily hydrolyzed by weak acid, liberating galactofuranose and oligosugar in dialyza-ble fractions.

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Using erythrocytes as antigen particles, number of antibody-forming cells was enumer-ated by immunocytoadehesion technique, in which formation of rosette was shown to be inhibited by anti-mouse immunoglobulin sera. This number increased in vitro after treatment of spleen cells of mice for 60min with RNA fraction extracted from spleen of mice immunized with erythrocytes used in the enumeration, and incubation of cells for 12 hr at 37C. Response of cells treated with immune RNA fraction was immunologically specific and was inhibited by puromycin or cycloheximide. The activity of immune RNA capable of converting nonimmune cells to antibody-forming cells was shown to be sensi-tive to ribonucleases but resistant to deoxyribonuclease and proteolytic enzyme.

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Digestion of rabbit streptococcal anti-M antibody with papain to produce univalent fragments resulted in the loss of its ability to fix complement. However, the digested univalent antibody still retained the indirect bactericidal activity in vitro, opsonic activity in vivo, and also the capability of protecting mice against the streptococcal infection. Thusly the biological activities of digested anti-M antibody appear to be similar to those of intact anti-M antibody.

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An investigation of the antibacterial action of decamethylenebis-(4-amino-5, 6, 7, 8-tetrahydroquinaldinium) acetate (THQ-Ac), a new antimicrobial compound, against Escherichia coli in comparison with the commonly available surface active agents was conducted. At high concentration, THQ-Ac inhibited synthesis of protein and nucleic acid in E. coli; its action, however, was not highly specific. Leakage of cellular constituents took place by the action of THQ-Ac but the amount of leakage was not as great as that induced by the same concentration of cetyl trimethylammonium bromide (CTAB). Cationic agents, such as CTAB and benzalkonium chloride, caused the lysis of the spheroplast of E. coli, whereas THQ-Ac did not. Additionally, adsorption of THQ-Ac to the cell-wall of E. coli also was investigated. According to our results, the site of action of THQ-Ac was considered to be the surface of bacterial cells, and it seemed to possess somewhat different activities from the other surface active agents.

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The pathomorphological changes and the multiplication process of varicella-zoster virus was studied in VERO cells infected with this virus isolated from the vesicle contents of varicella patient by Ogiwara and Nakamura [4]. The light microscopic observation of the virus infected VERO cell sheets showed at first the localized groups of cells which were round in shape and reflected the light strongly. These localized pathological changes gradually extended to the neighboring cells and finally such changes could be seen practically in the cells of the entire sheet. In the electron microscopic observation, the site where the pathological changes by varicella-zoster virus infection observed at first in the cell was the inside of the nucleus. Namely, the core particles of 70-90 mμ in diameter were made in the nucleus. And these core particles passed through the inner nuclear membrane, and then these particles in the vesicle of the outer nuclear membrane moved into the cytoplasm. The capsids of virus particles measured 90-110 mμ in diameter were made on the outside of core in the cytoplasm, and the thick membrane of the electron dense substances measured 190-220 mμ in diameter enveloped the virus particles with capsids. This membrane is one part of the envelope of virus particle. The mature virus particles are released from the host cell through the cell wall to the outside of the cell.

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The cell division of a strain of Bacteroides convexus was examined by the ultrathin sectioning and the electron microscopy. The cell division was initiated by the invagination of the cytoplasmic membrane from the opposite sites at the middle of the cell. The constriction of the cell wall also occurred simultaneously or soon after the initiation of the invagination of the cytoplasmic membrane. A short septum structure similar to those of gram positive bacteria originated within the base of mesosome. The two mesosomes arising from the opposite sites fused at the center of the cell. After the tips of invaginating outer membrane reached to the middle between cell center and original outer membrane, the mesosomes were reduced gradually and finally disappeared. In this stage of the cell division, a transverse septum was usually completed. The invagination of the outer membrane proceeded progressively and finally fused at the center of the division plane.

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Cultures of Nocardia and Streptomyces were grown on agar film on glass slides and the mode of breaking up of their vegetative mycelium was studied. It was found that they broke into shorter filaments by two modes: fragmentation and autolysis. There were two types of fragmentation: one is when the divided cells are separated by empty splits and the other is when the cells are not separated but connected to each other, no splits thus being formed. When autolysis occurred at small intervals along the hyphae and then the intervening cytoplasm disappeared, the interstices appeared as empty splits which could not be distinguished from those caused by fragmentation. With the progress of autolysis, parts of the vegetative mycelia became ghost-like or beaded, which parts usually remained for a long time in Streptomyces cultures but tended to disappear in Nocardia isolates. The splits which had been caused by fragmentation or by autolysis were sometimes filled with newly developed filaments. Although both fragmentation and autolysis were found in Streptomyces as well as Nocardia, the former was more abundantly observed in Nocardia and the latter was predominant in Streptomyces.

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Attempted virus isolation from fecal specimens using four different cell cultures in parallel: primary human embryonic kidney (PHEK) cells, human embryonic lung diploid (DHEL) cells, human cancer HEp-2 cell line and African green monkey kidney VERO cell line. Of 266 rectal swabs obtained from children, ages 0 to 11 from a small town in northern Honshu during August 1966, 74 (27.8%) yielded viral isolates. These included 58 coxsackievirus B3, 7 coxsackievirus B4, 3 echovirus 6, 3 adenoviruses, and 3 unidentified isolates. Of special interest is that practically no clinical disease was associated with this evident outbreak of coxsackievirus B3 infection. Our results provide evidence that HEp-2 and VERO cells are most sensitive, PHEK cells are less sensitive, DHEL cells are the least sensitive for isolation of coxsackievirus B3, isolation rates were 19.2, 12.8, and 7.5%, respectively. Sensitivity differences were revealed by time of appearance and rapidity of progress of the cytopathic effect, and the infectious titer attained in the passages of the isolates.

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A direct injection of various culture supernatants into a column of Porapak Q was investigated as a rapid and simple technique for gas chromatographic analysis of volatile metabolites produced by microorganisms. The usual metabolites such as methanol, ethanol, formic acid, 2-propanol (or acetone), 1-propanol, acetic acid, diacety1, 1-butanol, propionic acid, acetoin, butyric acid, 2, 3-butanediol and lactic acid were detected with-out any pretreatment. It was demonstrated that most strains of Bifidobacterium bifidum (Lactobacillus bifidus) produced both acetic acid and lactic acid which contrasts with re-sults from Escherichia coli and other Lactobacilli.

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The antigenic mannan of Candida albicans was degraded by acid-hydrolysis and the resultant oligosaccharides were fractionated by a carbon-Celite and a subsequent cellulose-powder chromatography to yield four oligosaccharides, pentaose, hexaose, heptaose and octaose, which involved 2, 6-di-0- and 6-0-substituted mannopyranosyl residues as the common feature. These oligosaccharides showed lower precipitation-inhibition activity than that of the hexaose of acetolysate, the strongest inhibitor among the oligosaccharides described in the preceding study. The order of inhibitory powers of oligosaccharides was as follows: hexaose of acetolysate>heptaose>pentaose=octaose>hexaose. The μmoles requiring for 50%-inhibition were 0.025, 0.15, 0.20, 0.20 and 0.50 respectively. The results clearly indicate that the determinant groups of the mannan of C. albicans em-ployed this study are the hexaose moieties which constitute the branching parts of polysaccharide.

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A sensitive assay system of Yaba virus (YV) was established in a cynomolgus monkey kidney cell line, JINET, in which the virus caused multilayered cellular foci countable even with the unaided eye. The specificity of the foci induced by YV in these cells was demonstrated by (1) the focus-forming ability was destroyed by heating at 60 C for 12 min; (2) the focus formation was inhibited by specific antiserum; (3) specific fluorescence was detected only in cells composing the foci when tested by fluorescent antibody technique; (4) a linear relationship was observed between the virus concentration and the number of foci formed; (5) YV preparation passed 20 times in JINET cells still possessed "tumorigenicity" in cynomolgus monkeys. The sensitivity of JINET cells to YV was comparable to that of cynomolgus monkeys, and YV was successively propagated in JINET cells with 2 log increase in infectivity titer during over 40 serial passages. Application of this assay system to growth kinetic studies of YV and quantitation of neutralizing antibody to YV is also discussed.

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