bluuebunny/crossref_metadata_2025 huggingface dataset:
Fine Structure of Strictly Anaerobic, Non‐Spore‐Forming, Gram‐Negative Bacilli
Authors: Ushijima Tsutomu
Published: November 2013

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.
Comparative Sensitivity of Various Tissue Cultures for Isolation of Coxsackievirus B3
Authors: Kawana Rinji , Matsumoto Ichiro
Published: November 2013

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.
Rapid Gas Chromatographic Analysis of Microbial Volatile Metabolites
Authors: Yoshioka Masanori , Kitamura Miyoshi , Tamura Zenzo
Published: November 2013

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, diacetyl, 1‐butanol, propionic acid, acetoin, butyric acid, 2,3‐butanediol and lactic acid were detected without any pretreatment. It was demonstrated that most strains of Bifidobacterium bifidum (Lactobacillus bifidus) produced both acetic acid and lactic acid which contrasts with results from Escherichia coli and other Lactobacilli.
Titration and Extensive Serial Passages of Yaba Virus In Vitro
Authors: Tsuchiya Yoshiki , Tagaya Isamu , Tsuruhara Takashi
Published: November 2013

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.
The Structure of Tobacco Mosaic Virus from Base Analogue Treated Tobacco Leaves
Authors: Nomura Akira , Taniguchi Takeshi , Hirai Tokuzo
Published: November 2013

Structure of TMV containing the base analogues such as 2‐thiouracil, 8‐azaguanine, and 5‐fluorouracil was studied. TMV containing these analogues had the same anion‐exchange chromatographic properties as had normal TMV, and 2‐thiouracil was incorporated into TMV particles, probably into TMV‐RNA. Cation‐exchange chromatographic properties of TMV containing these analogues had no significant difference as compared with those of normal TMV. TMV containing 2‐thiouracil was more stable to alkali degradation than normal TMV. Alkali‐degraded TMV containing 2‐thiouracil and normal TMV was characterized by chromatography. These results suggest that not surface structure but internal structure of TMV containing 2‐thiouracil was different from that of normal TMV.
Germination of B. anthracis Spores in the Peritoneal Cavity of Rats and Establishment of Anthrax
Authors: Hachisuka Yoetsu
Published: November 2013

Most of spores of B. anthracis introduced into the peritoneal cavity of rats could germinate when 5 mg of L‐alanine and 5 mg of adenosine were injected by the same route shortly before the spore injection. However the spores hardly germinated in untreated rats. To develop vegetative cells after spore germination, 1 ml of an aqueous solution containing 1% casamino acid and 1% of yeast extract was injected into the peritoneal cavity of rats before injection of 55×107 spores of B. anthracis. A few germinated spores and vegetative cells were seen in ascites, but their numbers were not enough to kill the rats. In order to produce a rapid germination and development to vegetative cells in a short time, 1 ml of solution containing 1% of casamino acid, 1% of yeast extract, 5 mg of L‐alanine and 5 mg of adenosine was injected before intraperitoneal administration of spore suspension. Spore germination and vegetative growth were tremendous and all of rats died within 24 hours. From these results it was concluded that the natural resistance of the rat for B. anthracis is partly due to unfavorable internal conditions for spore germination and subsequent vegetative growth.
Effect of Neonatal Thymectomy on Immunity of Mice against Salmonella Infection
Authors: Saito Kazuko , Takami Mamoru , Kawakami Masaya , Mitsuhashi Susumu , Fukai Kohji
Published: November 2013

Lowered peripheral lymphocyte count, reduced serum antibody response and partially reduced protection against infection with virulent strain of Salmonella enteritidis after immunization with live vaccine of S. enteritidis were observed in mice thymectomized at birth. However, the cellular resistance, which was observed in vitro in peritoneal macrophages from immunized animals, was not found to be reduced by neonatal thymectomy of mice. From these facts, the immunological dependence of cellular immunity on thymus in neonatal life was discussed.
Establishment of Candida albicans in the Alimentary Tract of the Germ‐Free Mice and Antagonism with Escherichia coli after Oral Inoculation
Authors: Nishikawa Takeji , Hatano Hitoshi , Ohnishi Nobuhiko , Sasaki Shogo , Nomura Tatsuji
Published: November 2013

To solve the problems of the multiplication and invasion of Candida albicans in the alimentary tract, germ‐free mice were orally inoculated with C. albicans and the antagonistic effect of superinfection with Escherichia coli was examined. C. albicans could easily be established in the alimentary tract of germ‐free mice by inoculation with less than 10 organisms, whereas in the alimentary tract of the SPF mice, having normal bacterial flora, C. albicans could not establish even after inoculation with an overwhelming dose. In germ‐free mice large numbers of the inoculated Candida were excreted in the feces throughout the observation period of 130 days, without affecting the conditions of the mice. By histopathological examination of these mice only one mouse showed microabscesses with leukocytic infiltration accompanied by hyperkeratosis and acanthosis in the epithelium of the forestomach. However even in this mouse no invasion of Candida cells into the acanthotic squamous cell layer was seen. After inoculation of the mice who had already E. coli in their gut as a monocontaminant with C. albicans, or even after inoculation of E. coli to the mice harboring Candida as a monocontaminant, E. coli always outnumbered C. albicans. In the former case Candida were even completely eliminated from the mice within few days. Thus, it appeared that in the alimentary tract of the mice, E. coli have the capacity to antagonize to C. albicans.
Cross Immunity of Mouse Mononuclear Phagocytes Induced by Immunization and by Transfer Agent
Authors: Saito Kazuko , Kurashige Satonori , Osawa Nobutaka , Kato Eisuke , Kawakami Masaya , Mitsuhashi Susumu
Published: November 2013

Peritoneal macrophages from mice, which had received an RNA fraction extracted from spleens of mice immunized with live vaccine of Salmonella enteritidis, were found to resist in vitro infection with heterologous species of Salmonella as well as the homologous one, but were susceptible to Mycobacterium tuberculosis. Cellular antibody demonstrated in peritoneal exudate cells from the immunized mice and from the immune RNA‐treated mice was shown to react to these Salmonella but not to M. tuberculosis. The relation of cellular resistance and antibody in the restricted cross‐protection of immunized mice in experimental salmonellosis is thusly demonstrated.
Development and Transduction of Rsistance to Novobiocin and Nikkol‐SNP 7.5 A (an Anionic Surfactant) in Staphylococcus aureus
Authors: Onodera Toshinao , Ishihara Keizo
Published: November 2013

The susceptibility to Novobiocin (NB) and Nikkol‐SNP 7.5 A (NS), an anionic surfactant, was studied in 458 strains of coagulase‐positive Staphylococcus aureus isolated from clinical sources. Twenty three (4.9%) of these strains were resistant to NB with minimum inhibition concentration (MIC) of 1.6 μg per ml or more, 97 (21.2%) resistant to NS with MIC of 6.25 mg per ml or more, and 17 (3.7%) resistant to both drugs. Cross‐resistance to NB and NS was found in 74 per cent of 23 NB‐resistant strains and 17.5 per cent of 97 NS‐resistant strains. Nearly one half of NS‐resistant strains belonged to phase group I, while the remainder were non‐typable. The majority of the NB‐resistant strains were not phage typable. In S. aureus strain PS 53 used for propagating phage 53, resistance to 25 mg per ml of NS was attained rapidly by single step without accompanying that to NB, whereas resistance to 25 μg per ml of NB developed gradually by three successive steps and was accompanied by a rapid development of resistance to NS first by two steps. The transduction experiments in strain PS 53 showed that resistance to NB and NS was jointly transduced and the genetic loci responsible for resistance to both drugs are closely linked.
In vivo Uncoating of Tobacco Mosaic Virus after Infection of Tobacco Leaves
Authors: Maghida Haruhiko , Hayashi Takaharu , Abe Tetsuya , Kiho Yukio
Published: November 2013

In vivo uncoating of tobacco mosaic virus (TMV) was studied. As Shaw had reported, initiation of uncoating reaction takes place very efficiently. Coat protein is removed from the virus as a peptide which is precipitable with trichloroacetic acid. Short rod particles with partly exposed RNA are thus formed. Further uncoating to coat protein‐free TMV‐RNA (28S) seems to take place with very low efficiency which is comparable to that of formation of local lesions on the inoculated leaf. From the data on the intracellular distribution of these products of uncoating reaction, mechanisms and significance of these reactions are discussed.
Isolation and Properties of Two New RNA Phages SP and FI
Authors: Miyake Tadashi , Shiba Tadayoshi , Sakurai Toshizo , Watanabe Itaru
Published: November 2013

New RNA phages were isolated from feces of Siamang Gibbon and infants, and their characters were investigated. These phages, SP and FI were serologically unrelated to any of known RNA phages (group I, II and III) and were classified into groups IV and V. Several other characters, such as filtration and elution patterns through membrane filters, buoyant densities in CsCl, and UV sensitivities of SP and FI were similar to those of the group III phages, suggesting that a certain similarity between group III and IV or V might exist. Peculiar phage‐host relations found in phages of FI group were also discussed. From these results, we propose new possible schemata for the grouping of RNA phages.
Determination of Pseudomonas aeruginosa by Biochemical Test Methods
Authors: Arai Taketoshi , Enomoto Seiji , Kuwahara Shogo
Published: November 2013

A new test method was devised for microbial gluconate oxidation, using an ammonium molybdate reagent. One loopful (about 2 mg wet wt.) of the test organism, grown on a nutrient agar plate for 18 hr, is transferred into 1 ml of the test liquid medium consisting of (NH4)2SO4 0.5 mg, potassium gluconate 10 mg, NaCl 5 mg, KH2PO4 2 mg, MgSO4·7H2O 0.1 mg, and 1 ml of distilled water, incubated at 37 C for 6 hr without shaking, and then mixed with 3 ml of 1% aqueous solution of ammonium molybdate and 0.2 ml of glacial acetic acid. The mixture is heated in boiling water for 5 min, followed by abrupt cooling with running water. A deep blue colour appears in a positive result. A total of 39 strains of Pseudomonas aeruginosa showed positive results by this method, whereas Aeromonas, Vibrio, Proteus group, Klebsiella, Citrobacter and Enterobacter A group were all negative. Though some strains of Enterobacter B group showed a weak blue colour, it could be easily differentiated from the deep blue colour of Pseudomonas. Longer incubation of test microbes in the test medium, and longer heating of the reaction mixture gave unsatisfactory results.
Relationship between Immunity and Interferon Production in Macrophages
Authors: Azuma Masanobu , Yamada Morihide , Nishioka Rikichi
Published: November 2013

The effect of antiserum on the interferon (IF) production in macrophages by Newcastle disease virus (NDV) has been studied. Completely neutralized NDV lost its IF‐inducing ability, but under‐neutralized NDV revealed an enhanced IF‐inducing capability. This enhancement of IF production by unneutralized NDV was also observed in antiserum‐treated cells. Enhancement phenomena appeared to be based on immunological specificity. Under‐neutralized NDV was also adsorbed more rapidly and at a higher rate to normal cells than unneutralized NDV. Unneutralized NDV also adsorbed more efficiently to antiserum‐treated cells and to cells derived from immunized mice than to normal cells. The significance of these phenomena is discussed in relation to the induction mechanism of IF in macrophages.
Genetic Properties of a T Factor and T‐r Recombinants
Authors: Kameda Mitsuo , Suzuki Mitsue , Nakajima Takashi , Harada Kenji , Mitsuhashi Susumu
Published: November 2013

Genetic studies have shown that one of the transfer (T) factors, T95, carries the following genetic properties: autonomous replication (rep+), conjugal transmission (tra+), transmission of the host chromosome (mat+), inhibition of F‐mating (ifm+) or fi+), inhibition of superinfection with R factor (irs−), and suppression of phage proliferation (spp−). An Escherichia coli K12 strain carrying T95factor is insensitive to male phages (f1 and f2). These facts indicated that the T95 factor is a sex factor but different from the F factors. By the interaction between T95 factor and the non‐transferable resistance determinants; r31(tet) for tetracycline resistance and r3 (kan) for kanamycin resistance, recombinants, T‐tet, T‐kan, and T‐tet‐kan, factors were obtained. These are capable of conferring drug‐resistance and are conjugally transmissible as a single genetic unit. E. coli K12 strains carrying T95 or T‐tet factor were insensitive to male phages and T‐tet (probably T95) factor was transmitted sexually at about 10−5 transmission frequency after 1 hr of mixed culture. On the contrary, E. coli K12 strains carrying T‐kan or T‐tet‐kan factor were found to become sensitive to male phages and their transmission frequency was 102 to 103‐fold higher than that of T‐tet factor. As a result of these findings, we concluded that the pili formation of T95 or T‐tet factor is in a repressed state but the formation of a recombinant between T95 (or T‐tet) and r3(kan) determinants results in the derepression of pili formation.
Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue
Authors: Inaba Yuji , Tanaka Yoshio , Ishii Sukemitsu , Morimoto Tomiaki , Sato Kunihiko , Omori Tuneyoshi , Matumoto Minoru
Published: November 2013

Replication of Ibaraki virus was not inhibited by 5‐iodo‐2′‐deoxyuridine, indicating that the virus is an RNA virus. The virus was resistant to ether, chloroform and deoxycholate, sensitive to trypsin, very labile at acidic pH but stable at pH 6.4 or higher, and was resistant to repeated freezing and thawing. The virus was readily inactivated at 56 C or higher, was fairly stable at 37 C, and very stable at 4 C, while it rapidly lost infectivity when stored frozen at —20 C. The virus was readily sedimented by centrifugation at 40 000Xg for 60 min. It readily passed through membrane filters of 200 mμ pore size, passed through 100 μfilters but only with some titer loss and did not through 50 mμ filters. In these tests, the bluetongue virus used as a control behaved in the same manner as Ibaraki virus. These findings provide additional evidence for the similarity of Ibaraki virus to bluetongue virus which had been previously demonstrated on the basis of seasonal incidence, symptomatology and pathology of the diseases caused by these viruses and the behavior of the viruses in cell cultures, embryonated eggs and laboratory animals. The present study, however, provided no evidence for any serological relation between these two viruses. More Information is needed to reach a final decision on the classification of Ibaraki virus, particularly regarding the morphology of the virion, the doublestrandedness of the viral RNA and other basic features.
Autolytic Enzyme System of Clostridium botulinum
Authors: Kawata Tomio , Takumi Kenji
Published: November 2013

Isolated cell walls of Clostridium botulinum type A strain 190L released an autolysin during autolysis of the cell walls. The autolysin was isolated from the cell walls, and partially purified 18.6‐fold by ammonium sulfate precipitation, chromatography on DEAE‐cellulose and gel filtration through Sephadex G‐100. The purified preparation of the autolysin showed 2 major and 2 minor protein bands on Polyacrylamide gel electrophoresis. Some properties of the autolysin were examined using SDS‐treated cell walls of the organisms as a substrate. The autolysin was active over a pH range of 6 to 8, with a maximum near pH 6.8. The lytic activity was stimulated by 10−4 M each of Co++, Mg++ and Ca++ in the order, whereas it was inhibited markedly by Cu++. Mercaptoethanol (10−4–10−3 M) significantly activated the lytic action. Trypsin and nagarse (10 μg/ml) also stimulated the lytic activity. The lytic spectrum of the autolysin toward the SDS‐treated cell walls obtained from various types of C. botulinum and C. perfringens indicated a relatively high specificity. After treatment with hot formamide the cell walls of C. botulinum increased in susceptibility to the autolysin.
Genetic Mapping of aro, pyr, and pur Markers in Klebsiella pneumoniae
Authors: Matsumoto Hideki , Tazaki Tadakatu
Published: November 2013

Various aro, pyr, pur, and two other markers were located on the genetic linkage map of Klebsiella pneumoniae by employing a mating system in which genetic material was transferred by conjugation. These markers are widely scattered on the chromosome, and their locations were found to be similar to those of Escherichia coli. Biochemical and genetic investigations suggested that in K. pneumoniae, as well as in E. coli, the biosynthesis of 3‐deoxy‐D‐arabino‐heptulosonic acid‐7‐phosphate was catalyzed by 3 isozymes. A revised genetic linkage map of K. pneumoniae including these markers has been presented.
Replication and Transfer of the R Factor in a Synchronized Culture of Escherichia coli
Authors: Yabe Yoriko , Mitsuhashi Susumu
Published: November 2013

According to mutation mapping after nitrosoguanidine treatment of successive samples obtained from a synchronized culture of Escherichia coli K12 carrying an R factor, it was found that the majority of mutations of the R factor were produced after a maximum peak of galactose mutant production. According to the order of the chromosome replication and the initiation point of the replication, these facts suggest that the replication of the R factor occurs after the replication of the galactose gene. The transfer frequency of the R factor from a synchronized donor culture to the nonsynchronized recipient was the highest after the replication of the chloramphenicol gene on the R factor, probably after the replication of the R factor genome. The recipient ability of the R factor in a synchronized recipient culture from a nonsynchronized donor was the highest after a maximum peak of the galactose mutation, and probably after the cell division in the recipient.
Decrease in Accumulation of Macrolide Antibiotics as a Mechanism of Resistance in Staphylococcus aureus
Authors: Yamagishi Saburo , Nakajima Yoshinori , Inoue Matsuhisa , Oka Yoko
Published: November 2013

A marked difference of levels of accumulation of macrolide antibiotics (Mac) was found between Mac‐sensitive (E642‐1 and 209P) and Mac‐resistant (MS642, MS57, U9, and 606) strains of Staphylococcus aureus by means of qualitative and quantitative bioassay methods. Though the levels of accumulation of the antibiotics in cells of Mac‐resistant strains were only one‐tenth that of the Mac‐sensitives, satisfactory evidence for a relationship between the accumulation of the antibiotics and the degree of sensitivity to those drugs was established in a physiological sense from a kinetic measurement of Ks and ν in the accumulation reaction of the antibiotics in the cells. The experiments with tritiated erythromycin or josamycin also support these results. It is suspected, by estimation of the accumulation in heated or UV‐irradiated cells, that levels of accumulation of the antibiotics in cells of S. aureus could be reflected by a binding affinity of their ribosomes for these antibiotics. There was no system for inactivating the antibiotics seen in the cells of the Mac‐resistants, i.e., MS642, 606, and U9.
Rapid Formation of Protoplasts of Streptomyces griseoflavus and Their Fine Structure
Authors: Sagara Yasunobu , Fukui Komei , Ota Fusao , Yoshida Nagayuki , Kashiyama Tatsuo , Fujimoto Michimasa
Published: November 2013

Glycine, at a concentration of 5–10%, inhibited the growth of Streptomyces griseoflavus. The lysozyme sensitivities of the cell walls of the organisms cultured with and without glycine were the same. However, lysozyme caused 40% lysis of whole cells from cultures with glycine but had no effect on cells from cultures without glycine. The cell walls of organisms grown with and without glycine contained glucosamine, galactosamine, muramic acid, lysine, glutamic acid, glycine, alanine and DAP. Lysine and DAP were present in equimolar amounts. Very small amounts of asparagine, threonine and serine were found in both cell walls. Protoplasts were obtained in a high yield by treating cells from cultures containing glycine with 10mg/ml of lysozyme in 0.01 M phosphate buffer (pH 7.0) containing 0.1 MgSO4 and 1 M sucrose for 3 hr. The protoplasts were spherical and were surrounded by cytoplasmic membranes. The nucleoplasm appeared less dense with a clear network of nuclear fibers and mesosomes, abundant in whole cells, were scarcely seen. Rod‐shaped forms without cell wall materials were sometimes observed under the electron microscope.
Transfer Agent of Immunity
Authors: Mikami Hirokazu , Kawakami Masaya , Mitsuhashi Susumu
Published: November 2013

Ribonucleic acid fractions extracted from spleens of immunized mice were shown to convert non‐immune spleen cells to antibody‐forming cells. RNA fractions which possessed the converting activity appeared in the spleen 3 to 5 days after immunization. Sucrose density gradient separation of crude RNA revealed that most of the active materials were in the 8 to 12S fraction and not in other fractions. In methylated‐albumin‐Kieserguhr‐column chromatography, the activity was also recovered in a limited molecular size range. Specific activity of the RNA fractions was increased by purification procedures.
Enhancement of 32P Incorporation into Phospholipids in Cultured Cells by Sendai Virus of Parainfluenza Type 1
Authors: Shibuta Hiroshi , Yamaura Koichi , Hirano Koichi , Matumoto Minoru
Published: November 2013

Sendai virus infection induced enhancement of 32P incorporation into phospholipids in chick embryo, monkey kidney and bovine kidney cells, as previously observed in chorioallantoic membranes of chick embryos. These findings indicate that phospholipid synthesis is enhanced upon Sendai virus infection. Ultraviolet irradiation abolished the ability of the virus to induce the enhanced synthesis of phospholipids, a fact suggesting that the phenomenon depends upon infectivity of the virus. Gamma irradiation of host cells little affected the enhanced 32P labeling of cellular phospholipids, suggesting that the function of host cell DNA may not be directly involved in the phenomenon.
Drug Resistance of Staphylococci
Authors: Kono Megumi , O'Hara Koji , Nagawa Masatoshi , Mitsuhashi Susumu
Published: November 2013

Induction of chloramphenicol (CM) resistance in Staphylococcus aureus was investigated using 166 CM derivatives and analogues. It was found that 18 compounds of the samples used were able to induce resistance to CM in Staphylococcus aureus S1477 harboring an inducible CM‐resistance determinant. Most of the samples which had a nitrophenyl moiety and a D‐ or DL‐threo isomer in its steric configuration were found to have inducer activity for CM resistance. Competent inducers are D‐isomer CM derivatives which have another substituent in place of the hydroxyl group at carbon atom 3 in the propanediol of the drug.
Biochemical and Immunological Studies on Aspergillus
Authors: Azuma Ichiro , Kimura Hamako , Hirao Fumio , Tsubura Eiro , Yamamura Yuichi , Misaki Akira
Published: November 2013

Two kinds of polysaccharides, glucan and glycopeptide (galactomannan‐peptide), were obtained from Aspergillus fumigatus (IFO 5840) by extraction with 50% pyridine and were purified by fractional precipitation with acetone, by a column chromatography on Dowex‐50 and DEAE‐cellulose and by the gel filtration on Sephadex G‐50 and G‐200. The glycopeptide, designated APSK‐66 fraction, showed both an Arthus and delayed type (tuberculin type) skin reactions in sensitized rabbits and guinea pigs. By treatment with proteolytic enzymes, the delayed type skin reactivity of APSK‐66 fraction was reduced but the Arthus type skin reactivity was not affected. However, the Arthus type skin reactivity of APSK‐66 fraction was completely lost by periodate oxidation, and the delayed type skin reactivity of APSK‐66 fraction was retained. The APSK‐66 fraction showed precipitation, complement fixation and passive hemagglutination reactions with rabbit antisera against A. fumigatus. Glucan, designated as APSK‐33 fraction, did not show any immunological activity when tested in the present experiment. The chemical structures of the glucan and galactomannan were discussed.
Genetic Studies of Kanamycin Resistance in Staphylococcus aureus
Authors: Chin Yiu‐Man , Harmon Shirley A.
Published: November 2013

Attempts to isolate directly kanamycin resistant transductants in Staphylococcus aureus were unsuccessful, although genes determining heme synthesis could be transduced. Use of the replica plating method, however, gave evidence that kanamycin resistance had been co‐transduced along with the genes controlling the production of heme‐containing enzymes of the electron transport system. In the presence of kanamycin, bacteriophage was unable to produce plaques on phage‐susceptible cells, and this antiphage effect of kanamycin was probably a reason why the direct transduction of kanamycin resistance was not detected, although many attempts and variations in the procedure were made. Treatment of a kanamycin resistant culture of S. aureus with acridine orange or ethidium bromide indicated that kanamycin resistance was conferred by a chromosomal factor.
Double Infection, Recombination, and Segregation of Two R Factors, R 100 and Rts1, and Their Possible Bearing on the Genetic Structure of R 100
Authors: Yoshikawa Masanosuke , Sakai Katsuko
Published: November 2013

Applying the observation by Yokota et al (1969) that a cell doubly harboring an R factor (R100) and a temperature sensitive R factor (Rts1) produces segregant R factors with various resistance patterns, a total of 271 segregant R factors were obtained. There were 163 resistant to (sul, str, kan), 39 resistant to (sul, str, cml, kan), 62 resistant to (sul, str, tet, kan) and finally 7 resistant to (tet, kan). More than 90% of the former 3 segregants were fi+ and the remainder, including all of the (tet, kan) segregants, were fi−. Some fi− segregants with the former 3 resistance patterns and all of the (tet, kan) segregants were nontransmissible. All of these segregants were still temperature sensitive. Based upon the results of three experiments; (a) the growth at 43 C to observe linked loss of the kan gene and the genes derived from R 100, (b) a conjugal analysis of the relevant resistant markers, and (c) a transductional analysis of these same markers, several conclusions were made. The 2 R factors both consisting of a circle were supposed to have recombined to form a larger circle which then further resulted in the final formation of smaller circles. The possible bearing of these observations and conclusions on the genetic structure of R 100 was discussed.
Purification and Properties of Dihydrostreptomyein‐Phosphorylating Enzyme from Pseudomonas aeruginosa
Authors: Kobayashi Fujio , Yamaguchi Masahito , Sato Junko , Mitsuhashi Susumu
Published: November 2013

The distribution of the dihydrostreptomycin (DHSM)‐phosphorylating enzyme was investigated using DHSM‐resistant strains of Pseudomonas aeruginosa, indicating that this enzyme was demonstrated from all of 7 DHSM‐resistant strains examined but not from a DHSM‐sensitive one. The DHSM‐phosphorylating enzyme was isolated from P. aeruginosa TI‐13 and purified about 205‐fold using Sephadex G‐75 and DEAE‐Sephadex A‐50 column chromatography. The optimal pH for the DHSM‐inactivation was around 10.0, and both adenosinetriphosphate (ATP) and Mg++ were required for the inactivating reaction. It was found that this enzyme inactivated only DHSM but not other aminoglycosidic antibiotics such as kanamycin, aminodeoxykanamycin, neomycin, paromomycin, lividomycin and gentamicin.
Studies of Drug Resistance and R Factors in Bacteria from Pond‐Cultured Salmonids
Authors: Aoki Takashi , Egusa Syuzo , Yada Chiyo , Watanabe Tsutomu
Published: November 2013

Considerable fractions of gram‐negative bacilli, Aeromonas liquefaciens, A. salmonicida, Pseudomonas, Hafnia and unidentified Enterobacteriaceae, isolated from the intestinal tracts of cultured Amago (Oncorhynchus rhodurus macrostomus) from 23 Amago‐ponds and of Yamame (O. masou ishikawae) from one pond as well as the water of 3 ponds in Shiga, Gifu and Nagano Prefectures and Tokyo Metropolis in Japan were found to be multiple‐drug‐resistant. Of these drug‐resistant strains, A. salmonicida and A. liquefaciens carried R factors at high frequencies and to be prevalent in many Amago ponds throughout Japan. These R factors had markers of resistance to SA, SA.TC, SA.SM.CM. All the R factors belonged to fi– type.
Bovine Respiratory Syncytial Virus
Authors: Inaba Yuji , Tanaka Yoshio , Sato Kunihiko , Omori Tuneyoshi , Matumoto Minoru
Published: November 2013

A large epizootic of an acute respiratory disease of cattle occurred in Japan during the months from October 1968 to May 1969. A virus was recovered in primary cultures of calf kidney and testicle cells from nasal swabs of affected cattle. Neutralization tests revealed the virus to be closely related to the Long strain of human respiratory syncytial virus. The virus induced cytopathic changes including the formation of syncytia and acidophilic‐cytoplasmic inclusions in calf kidney and testicle cell cultures. A calf inoculated with the virus by the respiratory route developed an illness resembling the natural disease. Most cattle clinically diagnosed as having the disease showed significant rises of neutralizing antibody titer for the isolated virus, whereas none or only small fractions of those animals showed serological evidence for recent infection with bovine ephemeral fever virus, infectious bovine rhinotracheitis virus, Ibaraki virus, bovine diarrhea virus, bovine adenovirus Type 7 and parainfluenza virus Type 3. Neutralization tests on paired sera revealed a wide dissemination of the isolated virus among cattle in many areas of the country during the epizootic. All these findings leave no doubt that the epizootic was caused by bovine respiratory syncytial virus. This is the first study that ever shows the presence of infection of cattle with this virus in Japan.
Cell Wall Synthesis in Staphylococcus aureus in the Presence of Protein Synthesis Inhibitory Agents
Authors: Nαkαo Masafumi , Kitanaka Eiryo , Ochiai Kazuyori , Nakazawa Shozo
Published: November 2013

Electron‐microscopic and biochemical studies on morphological changes in Staphylococcus aureus following exposure to protein synthesis inhibitory agents such as lincomycin (LCM), clindamycin (CLM), erythromycin (EM), and spiramycin (SP) are presented in this paper. It was demonstrated that bacterial cell walls became extremely thickened usually with the formation of multilayers, when exposed to each of the above‐mentioned antibiotics. Furthermore, electron density of the cytoplasm was higher in those cells exposed to drugs than in intact control cells. Incorporations of 14C‐labeled l‐lysine into the cell‐wall fraction and the protein fraction were measured for biochemical elucidation of these phenomena. Labeled lysine was selectively incorporated into the cell‐wall fraction when the test organism was exposed to the respective antibiotics. Uptake at 15 min after exposure was about twice as large as that of intact control cells. SP and CLM inhibited protein synthesis while they stimulated cell‐wall synthesis. The evidences for thickening of, and formation of multilayers in the bacterial cell walls following exposure to drugs were closely related to the stimulating action of these antibiotics on the cell‐wall synthesizing system. Morphology of resistant clinical isolates following such antibiotic exposure was also investigated using two staphylococcal strains, one resistant to EM alone and the other completely cross‐resistant to all the macrolides.
Serological Survey of Rhinovirus Infections in Japan
Authors: Kawana Rinji , Matsumoto Ichiro , Imai Takenobu
Published: November 2013

Serological evidence for dissemination of rhinovirus serotypes 3, 5, 13, 14, 17, 26, 30 and 41, and a Japanese isolate 149N, which has not been typed as yet, was obtained by measuring neutralizing antibody in a total of 750 serum samples collected in Iwate Prefecture, a northern part of the main island Honshu of Japan, in 1965 to 1969. Judging from the age‐specific antibody incidence curves obtained by testing sera from outpatients at Iwate Medical College Hospital, the circulation of types 13, 14, 30 and 41 seems more active than that of types 3, 5, 17 and 26. Comparison in the age‐specific antibody incidence between two groups of sera collected from outpatients at the hospital over different periods, October 1968–June 1969 and February–August 1968, revealed very active circulation of types 13, 14 and 30 over the period between the collections as compared with type 41. Some differences in the age preference were shown among the serotypes. The results of tests in different cities showed marked differences in the antibody distribution by age depending upon the place and serotype. Seroconversion was shown in some individuals who were tested one year apart. These findings are interpreted to indicate that there is a constant flux in the serotypes infecting any given population, and that several serotypes are usually circulating almost simultaneously.
The First Successful Isolations and Identification of Yersinia enterocolitica from Human Cases in Japan
Authors: Zen‐Yoji Hiroshi , Maruyama Tsutomu
Published: November 2013

From March 1971 through February 1972 isolation attempts of Yersinia enterocolitica were made on 442 specimens, consisting of 292 feces from patients hospitalized with symptoms of suggestive dysentery, 22 from sporadic cases of diarrhea in children, 43 specimens of ileum resected from autopsy cases which exhibited pathological changes in their intestine, and 85 appendix vermiformis which had been excised. A simultaneous survey on the normal distribution of the bacterium in healthy persons was conducted on 30587 fecal samples from adults and 402 specimens from newborns. As a result of the survey, 12 strains of Y. enterocolitica were isolated; two strains from the ileum of two autopsy cases, five from excised appendix vermiformis, two from feces of hospitalized patients suspected of dysentery, and three from feces of sporadic cases of diarrhea in children. No strains of this bacterium were isolated from healthy adults and newborns. Major pathological findings of autopsy cases were conspicuous hemorrhagic necrosis and edema of the small intestine, limited to within 1 meter of the ileocecum. Biotype and serotype of 12 isolates were as follows; one isolate from an excised appendix vermiformis was biotype 2 and serotype 5B, while all of the remaining 11 were biotype 4 and serotype 3. These results suggested that strains of a specific biotype and serotype were likely to be playing a very important role in the development of this illness in man.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Yamada Hiroyasu , Muraoka Shizuko , Takeya Kenji
Published: November 2013

The production of anti‐hapten antibody after immunization with trinitrophenylated (TNP) hamster erythrocytes (HRBC) or sheep erythrocytes (SRBC) was determined in high‐ and low‐responder mouse strains against HRBC antigen. 1) Anti‐TNP antibody was detected in sera of high‐responder DDD and CF1 mice after primary immunization with TNP‐HRBC, but not in those of low‐responder C57BL/6 mice. 2) Anti‐TNP antibody was detectable in sera of all the strains after primary immunization with TNP‐SRBC. 3) Production of anti‐TNP antibody was elicited after a booster injection of TNP‐HRBC in low‐responder C57BL/6 mice pre‐sensitized with HRBC in Freund's complete adjuvant. These results suggest that functions of thymus‐derived cells specific for HRBC antigen are deficient in low‐responder mice.
Comparison of the Chemical and Immunological Properties of the Pili of Three Types of Corynebacterium renale
Authors: Kumazawa Norichika , Yanagawa Ryo
Published: November 2013

Pili of Corynebacterium renale strains 115 (type I) and 42 (type III) were purified, and compared chemically and immunologically with those of strain 46 (type II). Purification of the pili of the two strains was more complicated than that of the pili of strain 46. Pili of the three types of C. renale were dispersed in 1mM ATP and were thermostable. Stability in antigenicity and morphology in 0.1n HCl and 0.1n NaOH was different, and the antigenicity of the pili was different among the pili of the three types of C. renale.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Nakashima Izumi , Ohta Fujito , Kobayashi Takashi , Kato Osamu , Kato Nobuo
Published: November 2013

Study was made to clarify the experimental conditions for the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) to exhibit maximum adjuvant effect on antibody production to bovine serum albumin (BSA) in mice. To obtain the maximum primary antibody response and also the strongest priming for a secondary response to BSA, 1000 μg of CPS‐K had to be injected intramuscularly into the same or adjacent site of BSA injection within the period of 1 hr before to 6 hr after the BSA injection. The optimum amount of BSA giving the maximum antibody response and also the strongest priming under these experimental conditions was 15 mg. In mice thus primed, an extremely high secondary response was induced by injecting 0.5 mg of BSA 30 days after the initial injection. The minimum amount of CPS‐K, to exhibit a strong adjuvant action, was 100 μg, which was equal to the minimum amount to induce immunologic paralysis to a homologous antigen. Extremely large amounts, such as 100 to 300 mg per mouse of BSA, were also strongly immunogenic when injected together with paralyzing doses of CPS‐K. In vitro admixture of BSA and CPS‐K before injection did not strengthen adjuvant action of CPS‐K. Alum‐precipitated BSA mixed with CPS‐K was not more immunogenic than native BSA mixed with CPS‐K. Addition of Freund's complete adjuvant to an injection of BSA and CPS‐K mixture did not enhance the adjuvant effect of CPS‐K.
Slime Production by Pseudomonas aeruginosa
Authors: Goto Sachiko , Murakawa Takeo , Kuwahara Shogo
Published: November 2013

Using Pseudomonas aeruginosa culture IFO 3445, the nutritional requirements and cultural conditions suitable for slime production were investigated. A synthetic medium was established from the experimental results, which was composed of sodium glutamate, glucose, phosphate and magnesium salt. When a cellophane plate method was used, incubation at 37 C for 3 days attained the highest relative viscosity. In the presence of an oxidizable carbohydrate the relative viscosity of the culture fluid was reduced with the acidic reaction, and recovered if the reaction was adjusted to pH 7–8.
Characteristics of Group A Streptococcal Variants Treated with Mitomycin C
Authors: Hirano Toyo , Itoh Takako , Tomura Tsuneko , Yoshioka Morimasa
Published: November 2013

Transfers of Streptococcus pyogenes strain T12 in Todd–Hewitt broth containing stepwise increases in amounts of mitomycin C (MC) gave rise to slight changes of their colonial appearances. Variants thus obtained were examined for antibiotic and bile resistances; production of streptolysin‐S, ‐O and deoxyribonuclease; growth in alkaline medium, high salt concentration, and at 10 C and 45 C; sugar fermentations, and precipitin reactions. Four strains retained group A antigen, but some of them lost the ability to produce hemolysins and deoxyribonuclease, and acquired resistance to bile, penicillin and streptomycin as well as MC, and to physical environments. Four other strains lost group A antigen and acquired new antigens common to cells of group C, group D, or highly antibiotic‐resistant mutants reported previously. A variant which reacted with group C antiserum contained galactosamine, but not glucosamine, while the parent strain showed the reverse pattern. Many other variants contained both hexosamines. Even a variant, strain TL3‐2, reacted strongly only with group A antiserum, but contained glucose and both hexosamines. These strains having galactosamine possessed uridine diphosphate (UDP)‐N‐acetylglucosamine‐4‐epimerase activity which converted the substrate into UDP‐N‐acetylgalactosamine, while the parent strain failed to demonstrate the existence of this enzyme. The variants were discussed with respect to the group A streptococcal variations possessing no more original characteristics.
Studies on the Neutralization of Herpes Simplex Virus
Authors: Yoshino Kamesaburo , Kishie Taka
Published: November 2013

Herpes virus was reacted with an early rabbit antiserum containing predominantly complement‐requiring neutralizing (CRN) antibody to produce CRN‐antibody‐sensitized virus (SV), and the action of complement (C') upon SV was studied. Reduction of infectivity due to C' was about equal with undiluted and 1000‐fold diluted SV. Even higher dilutions which contained about 10 to 100 infectious units per 0.05 ml were also completely inactivated by C'. Kinetic experiments revealed that the velocity of titer reduction in the presence of C' of 100‐fold diluted SV was not slower than that of undiluted SV. When SV was first treated with C' and then diluted 100‐fold, the surviving virus showed but a slightly reduced efficiency of filtration through the 0.45 μ Millipore membrane as compared with SV first diluted 1: 100 and then treated with C'. The titer reduction of SV–C'1 complexes in the presence of C'4 followed a one‐hit curve. These results indicated that the reduction of infectivity of SV due to C' was not a result of immunoaggregation of infectious SV. Alternative possible mechanisms of the action of C' are discussed.
Field Trial with a Further Attenuated Live Measles Virus Vaccine
Authors: Makino Satoshi , Sasaki Keiko , Nakamura Naoyoshi , Nakagawa Masaharu , Kasahara Shiro , Mizunoe Kimifusa , Hashimoto Takashi , Ebihara Tutomu , Nakazawa Susumu , Oka Hide , Sato Hazime , Watanabe Osamu , Niino Kenji , Koga Fumiwaka , Onuma Masayuki , Kagami Kazuyuki , Kimura Mikio , Ishiguro Yoshio , Yoshida Zengi , Fugii Hiroshi
Published: November 2013

A live measles virus vaccine was prepared with a virus strain attenuated by passage in the primary sheep kidney cell culture. All of 209 initially seronegative children showed seroconversion after subcutaneous inoculation of one dose of the vaccine. The geometric mean of neutralizing antibody titers following vaccination was 26.6. Axillary temperature of 37.5 C or higher was recorded in 23.6% of 199 vaccinees with adequate clinical records, but only 4% had fever over 39.0 C. The average maximal temperature in feverish children was 38.3 C. Fever usually lasted for only one or two days. Neither febrile convulsion nor other toxic reactions was observed.
Ribonucleic Acid Dependent Ribonucleic Acid Replicase in the Immune Response
Authors: Saito Kazuko , Mitsuhashi Susumu
Published: November 2013

An immune ribonucleic acid (iRNA) preparation was made using phenol extracts of spleens of mice previously immunized with Salmonella tennessee flagella. An enzyme, also prepared from the spleens of these mice, induced the incorporation of 3H‐UTP into the acid‐insoluble fraction in a cell‐free system in the presence of this RNA. The enzyme activity could be demonstrated from the spleens of immunized mice but not from normal ones, and this activity was also inhibited by two derivatives of rifamycin. Treatment with ribonuclease or heating at alkaline pH resulted in a loss of activity in added RNA. The 3H‐uridine‐labeled product was found resistant to ribonuclease treatment but became sensitive when the product was subjected to heat treatment. However, actinomycin D, mitomycin C or bleomycin A2 did not inactivate the enzyme activity. These results suggest that this enzyme induces the incorporation of UTP into the acid‐insoluble fraction using iRNA as a template and the product may be a newly synthesized RNA which forms a hybrid with iRNA. This enzyme activity may play a role in the antibody formation process, and may account for the in vivo replication of iRNA by this enzyme, viz., probably an RNA‐dependent RNA replicase.
Japanese Quail Embryo Cells as a Host of Group A Arboviruses
Authors: Takayama Naohide , Simizu Bunsiti
Published: November 2013

Japanese quail embryo (QE) cells were compared to chicken embryo (CE) cells with regard to the production of some group A arboviruses. The virus yield in QE cells was about the same as that in CE cells. The sensitivity of plaque assay of the viruses in QE cells was nearly equal to that in CE cells, although a certain range of fluctuation among individual culture bottles of QE cells was observed. QE cultures were found to possess some advantages for production and titration of group A arboviruses when compared CE cell system.