bluuebunny/crossref_metadata_2025 huggingface dataset:
Recombinants of a Defective Lysogen of Staphylococcal Strains
Authors: Wentworth Berttina B. , Romig William R.
Published: November 2013

When certain staphylococci are lysogenized with serotype A phages, a resident defective prophage is dislodged. The extent of the genome of this resident prophage was examined by studying the progeny from recombination between it and an invading phage. The recombinants appeared to have acquired several characteristics from the defective lysogen, suggesting that the resident prophage contains genetic regions other than that specifying immunity.
Multiplication of Tobacco Mosaic Virus in Suspension Culture of Tobacco Cells
Authors: Motoyoshi Fusao , Oshima Nobuyuki
Published: November 2013

The present study was attempted to establish an in vitro system for TMV infection and multiplication. A friable cell line Bf1 was isolated from an original callus line which which obtained from a pith of Nicotiana tabacum. Cells of the line were stirred with ordinary TMV by means of a grass rod to be mechanically infected, suspended in the liquid medium, and incubated on a reciprocal shaker. Infectivity of TMV in the cells was determined by the local lesion assay with the leaves of N. glutinosa. It was increased in the incubated cells, reflecting that TMV was able to multiply in the in vitro system. An exponential increase of infectivity was observed during 2 and 5 days after inoculation.
Transfer Agent of Immunity
Authors: Kawakami Masaya , Kitamura Kouichi , Mikami Hirokazu , Mitsuhashi Susumu
Published: November 2013

Using erythrocytes as antigen particles, number of antibody‐forming cells was enumerated 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 60 min with RNA fraction extracted from spleen of mice immunized with erythrocytes used in the enumeration, and incubation of cells for 12 hr at 37 C. 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 sensitive to ribonucleases but resistant to deoxyribonuclease and proteolytic enzyme.
Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue
Authors: Omori Tuneyoshi , Inaba Yuji , Morimoto Tomiaki , Tanaka Yoshio , Kono Michi , Kurogi Hiroshi , Matumoto Minoru
Published: November 2013

Several strains of a virus, designated as Ibaraki virus, were isolated in bovine cell cultures from natural and experimentally produced cases of the disease. In some of these cases, antibody titer rises were shown against Ibaraki virus. The clinical materials used for virus isolation induced in calves a mild illness similar to the natural disease, as well as neutralizing antibody against Ibaraki virus. Antibody titer rises were also shown in a high percentage of natural cases clinically diagnosed as having the disease. Antibody was not detected in any of the cattle in non‐epizootic areas, while many cattle in the epizootic areas had antibody after the outbreak. In the epizootic areas the antibody incidence after the outbreak was much higher among cattle diagnosed as having the disease during the outbreak than those not affected. These results leave little doubt that Ibaraki virus is the etiologic agent of these disease. Ibaraki virus became less virulent for calves upon serial passage in bovine cell cultures. The bovine cell cultures appeared less sensitive than the calf inoculation for the primary detection of the virus. The use of intermediary calf inoculation before inoculation into tissue culture may improve the isolation rate, but the tissue culture technique itself should be improved as calf inoculation is both expensive and cumbersome.
Antigenic Structure and Phase Variation in Bordetella pertussis
Authors: Nakase Yasukiyo , Takatsu Kuniyoshi , Kasuga Tadayoshi
Published: November 2013

Relationship between K agglutinogenic factors of Bordetella pertussis were analyzed by agglutinin‐adsorption tests. Factors 1 and 3 corresponded to L agglutinogen, while 2 and 4 corresponded to S agglutinogen. The experiment also indicates that B. pertussis phase I strains belong to a single serotype, while types 1–2–3, 1–2, 1–3 and 1 strains belong to the author's intermediate phase which probably arose from phase I parents. Strains isolated by us in 1952 to 1953 were type 1–2–3–4 which possessed both L and S, but a few strains isolated in 1962 and 1966 were type 1–3. The occurrence of such a defect in antigens was discussed. A single phase I vaccine seemed to give enough protection against various serotypes of pertussis organisms.
Unstable Mutants of R Factor
Authors: Hashimoto Hajime , Iyobe Shizuko , Mitsuhashi Susumu
Published: November 2013

Thirty mutants sensitive to tetracycline were obtained from an R100 factor capable of conferring resistance to tetracycline (TC), chloramphenicol (CM), streptomycin (SM) and sulfanilamide (SA). Among the TC sensitive mutants, three showed a high frequency of spontaneous loss from host strains. The genetic loci governing the stability of R factor in host bacteria were denoted as stb. The stb– R factors have lost many of the properties of a wild type R factor, such as, the capability to sexually transfer drug resistance and host chromosome, to confer superinfection immunity and to inhibit F function. All of these properties did not revert to a wild type phenotype, suggesting that these mutations are deletions including genetic determinants governing both TC resistance and stability of R factor. Recombinational analysis between stb– and stb+ R factors indicated that crossovers between the stb loci and those governing CM (or SM.SA) resistance took place at high frequency. No crossovers were detected between stb loci and those governing TC resistance, indicating that the stb loci are linked closely to the loci governing TC resistance.
Heat‐Labile Toxin of Bordetella Pertussis Purified by Preparative Acrylamide Gel Electrophoresis
Authors: Nakase Yasukiyo , Takatsu Kuniyoshi , Tateishi Masayoshi , Sekiya Kachiko , Kasuga Tadayoshi
Published: November 2013

The heat‐labile toxin (HLT) of Bordetella pertussis was purified by column chromatography on DEAE‐cellulose, salt fractionation and preparative acrylamide gel electrophoresis. The toxin obtained was confirmed to be free from hemagglutinins, protective antigens, histamin sensitizing factors, and K and O agglutinogens, and was shown to be homogenous by agar gel diffusion tests, ultracentrifugation, and electrophoresis. The minimal necrotic dose of the HLT in guinea pig was 0.01 μg dry weight. It was a protein in nature, contained a sugar moiety and possessed S value of 1.4. The toxicity of purified HLT was increased by mixing with a protein, such as rabbit serum.
Chemical and Biological Properties of the Purified O Antigen of Bordetella pertussis
Authors: Nakase Yasukiyo , Tateisi Masayoshi , Sekiya Kachiko , Kasuga Tadayoshi
Published: November 2013

A cell wall fraction obtained by sonic oscillation, yielded a Bordetella pertussis O antigen which possessed the ability to both adsorb O agglutinins and to produce the O agglutinins in animals. It was purified by a phenol water extraction and centrifugation method. Physico‐chemical and biological properties of the O antigen were similar to that of the so‐called endotoxic lipopolysaccharide. It contained 2.6% nitrogen, 1.2–1.5% phosphorus, 9.0–11.0% heptose, 0.3% 2‐keto‐3‐deoxyoctonate (KDO), 13.3–13.4% as total carbohydrate, 16–16.5% hexosamine and 25–32% lipid. Homogeneity of the purified O antigen was confirmed by ultracentrifugation, electronmicrograph and gel‐diffusion tests. It possessed significant pyrogenicity, adjuvant effect on antibody production, and toxicity to animals, but not protective antigenicity.
Use of a Diffusion Barrier Technique in Bioautography
Authors: Nakamura Hiroshi , Samejima Keijiro , Tamura Zenzo
Published: November 2013

A diffusion barrier was used in bioautography to prevent the diffusion of compounds separated by chromatography or electrophoresis in a seeded agar toward the separation direction. Two main advantages were demonstrated in the present method; (1) better separation and identification of compounds occurring closely positioned, and (2) more sensitive detection owing to compact growth zones. The present method has proved satisfactory and reliable for the routine detection of unknown growth factors.
Morphology and Chemistry of the Bacterial Cell Wall
Authors: Ushijima Tsutomu
Published: November 2013

The location of the mucopeptide in the cell wall of Bacteroides convexus was determined by electron microscope after enzymatic and chemical treatment (papain, pepsin, lysozyme and phenol). In the five layered cell wall the innermost electron dense layer (or a part of it) proved to be the mucopeptide. The molar ratio of amino sugar and amino acid components of purified mucopeptide was about 1:1:1:1:1:1 for glucosamine, muramic acid, L‐alanine, D‐glutamic acid, DL(meso)‐diaminopimelic acid and D‐alanine.
Ibaraki Virus, an Agent of Epizootic Disease of Cattle Resembling Bluetongue
Authors: Matumoto Minoru , Inaba Yuji , Tanaka Yoshio , Morimoto Tomiaki , Omori Tuneyoshi
Published: November 2013

Ibaraki virus multiplied and induced cytopathic effects in primary cell cultures of bovine, sheep and hamster kidney and chick embryo, and cultures of BHK21‐WI2 cells of baby hamster kidney origin and mouse fibroblastic L cells, but did not in primary cultures of horse and swine kidney cells and HeLa cell cultures. The virus was readily passaged serially in 4 to 5‐day‐old eggs using the yolk sac inoculation and incubation at 33.5 C. The viral growth was better in eggs incubated at 33.5 C than 37 C, and in younger eggs, with high yields in yolk, yolk sac and embryo. The virus was passaged serially in newborn mice by the intracerebral route. The virus multiplied in the brain of mice of any age, but younger mice supported better viral growth and developed encephalitis. As the age of mice increased, the morbidity and mortality became lower, no deaths being observed in 2 to 3‐week‐old mice. These observations in cell cultures, embryonated eggs and mice emphasize the similarity of Ibaraki virus to bluetongue virus. No evidence was obtained that young adult rabbits and weanling guinea pigs are susceptible to Ibaraki virus. The virus seemed to have little if any pathogenicity but infectivity of a low grade for sheep, while the virus is capable of inducing clinical illness, even severe in some instances, in cattle. This is in contrast to bluetongue virus which is highly pathogenic for sheep and much less so for cattle. Serial passages in embryonated eggs and suckling mice resulted in attenuation for cattle of Ibaraki virus.
Comparison of Penicillinases Produced by R Factors Isolated from Ampicillin‐resistant Gram‐negative Bacteria
Authors: Ooka Tadaaki , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Among 9365 Shigella strains and 1435 other gram‐negative bacteria of clinical origin, 88 (0.9%) and 242 (16.9%) strains were resistant to aminobenzyl‐penicillin (AB‐PC). Of these 330 AB‐PC resistant strains, 138 (57%) transferred their AB‐PC resistance by conjugation. More than 50% of AB‐PC resistant Escherichia coli or Klebsiella strains carried R factors capable of conferring AB‐PC resistance. These R factors were transferred to E. coli K‐12 strains and after were examined for the levels of resistance to cephaloridine (CER) and 5 penicillins; (AB‐PC, penicillin‐G (PC‐G), phenethicillin, 6‐aminopenicillanic acid (6‐APA), and cloxacillin). Concerning PC‐G, AB‐PC or CER resistance, there were found 3 levels of drug‐resistance, labelled by us as low, middle and high. From the patterns of the resistance levels to 5 penicillins and CER, R+ strains were classified into 4 major groups. Group I strains were highly resistant to all of the penicillins tested and group II strains showed high resistance only to 6‐APA. Penicillinase (PCase) produced by group II strains was similar to that reported previously, and group I PCase showed almost the same substrate specificity but has 20 times higher specific activity as that of the group II PCase. In group III or IV, the resistance levels to some penicillins were low. Group IV PCase was different from that which has been thus far reported.
Macrolide Resistance in Staphylococcus aureus
Authors: Shimizu Mikio , Saito Tetsu , Mitsuhashi Susumu
Published: November 2013

It was demonstrated that spiramycin (SP)‐resistance could be related to the decrease in binding of ribosomes to SP and that the SP‐binding to ribosomes was related with inhibition of polypeptide synthesis by SP in a cell‐free system in staphylococcal strains. These facts were also observed in Mac (macrolide)‐inducible strains, in which resistance to Mac antibiotics is enhanced by prior treatment with subinhibitory concentrations of erythromycin. From these results, it was concluded that the mechanism of resistance to Mac antibiotics is accounted for by alteration of ribosomes in staphylococcal strains and that this alteration of ribosomes is caused not only by mutation but also by induction.
Isolation of R Factors Conferring Kanamycin Resistance from Tuberculous Patients Treated with Kanamycin
Authors: Hashimoto Hajime , Ike Yasuyoshi , Hosoda Tetsuji , Mitsuhashi Susumu
Published: November 2013

In a survey of 228 tuberculous patients who had received kanamycin (KM) treatment, KM‐resistant Escherichia coli or Escherichia freundii were isolated from 29. From the patients who had not received KM‐therapy, KM‐resistant strains were not isolated. Of the 29 KM‐resistant strains isolated, 5 E. coli strains were also resistant to other aminoglycoside antibiotics, i.e., paromomycin, fradiomycin and streptomycin. They were also resistant to chloramphenicol and sulfanilamide and all of these resistances could be transferred to E. coli strains by conjugation. The resistance was considered to be conferred by an R factor. The remaining 24 KM‐resistant strains were identified as E. freundii. In these level of KM‐resistance was rather low and conjugal transfer was not observed. All were resistant to paromomycin but sensitive to fradiomycin. When 381 KM‐sensitive strains were plated on an agar plate containing 25 μg of KM/ml, 23 KM‐resistant strains were isolated. The mutants thus obtained were similar to the second type of 24 KM‐resistant strains isolated from patients. Thus it was strongly suggested that the 24 strains carrying low levels of KM‐ and paromomycin‐resistance, that is, conjugally nontransmissible, appeared by spontaneous mutation in the intestines of patients who had received KM‐treatment.
Multiplication of Mycobacterium lepraemurium in Mouse Foot‐Pad Cell Culture
Authors: Matsuo Yoshiyasu
Published: November 2013

A limited multiplication of Mycobacterium lepraemurium was observed in cultures of mouse foot‐pad cells up to the third sub‐culture. After 96 hr of incubation at 33 C for phagocytosis, infected cells were washed to remove unphagocytized bacteria and transferred to new culture vessels. There was a 4.9‐fold increase in the number of bacteria during 70 days of successive cultures up to the third culture in flasks, giving an overall generation time of 30.4 days. Acid‐fast stained cover‐slips taken from the culture in Leighton tubes revealed intracellular multiplication of M. lepraemurium, frequently in bundles of hundreds of bacteria, and gave an average generation time of 16.4 days in the primary, 15.7 days in the secondary, and 8.1 days in the tertiary culture. Elongation of M. lepraemurium was observed as early as 6 days after infection. All attempts to grow the acid‐fast bacteria from the original bacillary suspension and the cell suspension at the end of the experiment on artificial culture media have failed. However, subcutaneous inoculations of these materials to mice produced the typical lesions caused by M. lepraemurium.
Reexamination and Characterization of the T‐Agglutination Complex or Pattern of Streptococcus pyogenes: Preparation of Anti‐T Factor Sera
Authors: Takizawa Kinjiro , Akiyama Shoichi , Miyamoto Yasushi
Published: November 2013

The T‐agglutination complex or pattern of Streptococcus pyogenes which cannot be strain typed by a single definite anti‐T monovalent serum but, because of the presence of a common T antigen, requires a set of two or more anti‐T sera. Of these complexes, strains belonging to the 3‐13‐B3264; 8–25‐Imp 19; and 5–27–44‐complexes from outside Japan were studied, and each could be typed as a definite T‐type. These discrepancies were attributed to the different procedures employed by us in adsorbing the original antisera. We observed the same patterns or complexes after complete removal, from our original antisera, of the non‐type‐specific or group‐specific antibodies by absorption. Consequently we postulate that these patterns merely represent cross reactions occurring in minor T antigens. Therefore we attempted in this investigation to improve the absorption procedures using a much larger amount of whole cells, to obtain separate factor sera. These were attained with little difficulty and the results given by these factor sera were clear cut. The present paper reports on the reexamination and characterization of the complex or pattern of the resulting T‐agglutination complexes.
Determination of Pseudomonas aeruginosa by Biochemical Test Methods
Authors: Arai Taketoshi , Otake Masako , Enomoto Seiji , Goto Sachiko , Kuwahara Shogo
Published: November 2013

A rapid and simple test method for the detection of acylamidase activity of Pseudomonas aeruginosa was devised. One loopful of a nutrient agar overnight culture of a test organism was inoculated into 1 ml of a test medium consisting of 0.2% KH2PO4, 0.01% MgSO4·7H2O, 0.5% NaCl and 0.1% acetamide (final pH 6.8). After aerobic incubation at 37C for 6 hr, one drop of Nessler's reagent was dropped into the test medium. A reddish‐brown sediment appeared immediately if results were positive. Of 40 test strains of P. aeruginosa 39 gave strongly positive results. A strain showed a weakly positive result after 6 hr incubation, but the reaction became stronger after 18 hr culture. Other species of Pseudomonas, and various species of bacteria such as genera Vibrio and Aeromonas, and family Enterobacteriaceae were negative in this test. From these experimental results, the acylamidase test was considered to be highly specific for strains of P. aeruginosa, and therefore useful as a reliable method for the identification of this species.
Polysomes Containing Infecting Viral Genome in Tobacco Leaves Infected with Tobacco Mosaic Virus
Authors: Kiho Yukio
Published: November 2013

This study concerns the interaction between parental virus RNA and host cell components in tobacco leaves infected with tobacco mosaic virus (TMV). By using a gentle method for disruption, extracts were obtained from tobacco leaves infected with 32P‐labelled TMV and the fate of infecting parental 32P‐TMV‐RNA was studied. Sucrose gradient centrifugation analysis showed that the parental 32P‐RNA distributed into four regions: free RNA, partially uncoated TMV, TMV and a structure heavier than TMV. The heavier structure was considered to be polysomes carrying the parental TMV‐RNA as messenger RNA based on its size, sensitivity to RNase, dependence of its formation on protein synthesis and the kinetics of its appearance after infection. Control experiments were done to exclude the possibility that the structure is not an artifact aggregate produced by an interaction between partially uncoated virus or its RNA and host cell components. Polysomes containing TMV‐RNA were found as membrane bound forms. Based on these data, it is suggested that uncoating of TMV and formation of polysomes are closely related and evidence has been obtained which suggests that uncovering of the viral genome and its translation takes place hand‐in‐hand on a single virion.
Factors Affecting the Germination of Spores of Clostridium botulinum Type E
Authors: Ando Yoshiaki , Iida Hiroo
Published: November 2013

Germination of spores of Clostridium botulinum type E under varying conditions of Eh, pH, temperature, activation, and nutrition was studied. Germination in a complex medium consisting of thiotone, glucose and bicarbonato occurred at any level of initial Eh, regardless of the poising procedures employed. On the other hand, postgerminative development was affected by the initial Eh in the germination medium. Under an anaerobic condition, germination, outgrowth and division of cells were characterized by a peculiar profile of an Eh‐time curve. The optimal conditions for germination of unheated spores in the complex medium were incubation at 37 C and pH 6.6. The rate of germination increased with ageing time, the maximum rate being attained after storing an aqueous suspension of spores at 4 C for 20 days. The optimal heat activation was indicated when subjecting spores to heat‐shock at 60 C for 10 min. Germination in a casamino acids medium was markedly stimulated by a supplementation of lactate. The minimum requirements of compounds needed for the germination of spores of C. botulinum type E were l‐alanine, glucose, lactate, and bicarbonate (CO2). Germination occurred only in the following combinations of these compounds: l‐alanine+glucose+lactate, l‐alanine+glucose+bicarbonate, or l‐alanine+lactate+bicarbonate.
Studies on the Antigenic Activities of Yeasts
Authors: Sunayama Hiroyoshi , Suzuki Shigeo
Published: November 2013

In order to provide further information about the immunochemical differences between two mannans of Candida albicans serotype A and serotype B, quantitative precipitation‐inhibition tests of anti‐C. albicans serotype B serum were carried out in the present study. Oligosaccharides were prepared by acetolysis of a homologous mannan, and a1→2 linked di‐, tri‐, tetra‐, penta‐ and hexasaccharide were separated in chromatographically homogeneous states. The latter two oligomers contained a small amount of an isomer containing a1→2 and a1→3 linkages. In the precipitation‐inhibition tests of anti‐C. albicans serotype B serum with its homologous mannan and heterologous mannan of C. albicans serotype A, the inhibitory power of the oligomers was of the following order; hexa‐> penta‐> tetra‐> tri‐> disaccharide, and the amounts for 50%‐inhibition of the former 4 oligomers were 0.02–0.03, 0.05–0.07, 0.1–0.2 and 0.3–0.4 μmoles respectively, whereas disaccharide was very poor inhibitor. The lower oligomers, a1→2 linked tri‐ and tetrasaccharide, showed considerably strong inhibitory activities. The results obtained in the present study confirmed that the antigenic determinants of the mannan of C. albicans serotype B is the hexasaccharide moiety corresponding to the longest branched chains of mannan, and moreover, the a1→2 linked tri‐ and tetrasaccharide moieties play an important factor in dominating immunochemical specificity.
Inhibition of Phage Growth by an Antibiotic Rugulosin Isolated from Myrothecium verucaria
Authors: Nakamura Shinichi , Nii Fusako , Shimizu Masanao , Watanabe Itaru
Published: November 2013

It was found that (−)rugulosin, an antibiotic isolated from Myrothecium verucaria, had a potent anti‐phage effect on RNA phages (MS2, GA and Qβ) and DNA phages (δA and T4). The effect was almost independent of the host bacterial strains used. By a detailed investigation using MS2, it was revealed that the antibiotic did not affect the free phage or host bacterium alone but inhibited phage multiplication, and the degree of the inhibition depended on the multiplicity of infection. The inhibition was not mainly due to a drop in the burst size but rather was due to a decrease of the phage‐producing cells during the early stages of phage infection and replication.
Electron Microscopic Study of Rhinovirus Replication in Human Fetal Lung Cells
Authors: Kawana Rinji , Matsumoto Ichiro
Published: November 2013

Human embryo lung cells infected with rhinovirus type 2 (HGP strain) were sampled at intervals during viral growth and thin sections examined by electron microscopy. Infectivity assay showed an increase of intracellular virus 7 to 8 hr postinoculation and a peak titer at 16 to 18 hr. Changes resulting from infection, as revealed by electron microscopy, comprised (i) the appearance in the cytoplasm, at 8 hr postinfection, of large ribosomal clusters, membrane‐enclosed bodies which proliferated extensively in the central zone of the cell, and viroplasm, aggregates of irregularly dense, coarse, granular material, and (ii) the appearance in the cytoplasm, at 16 hr postinoculation, of recognizable progeny virus particles. The particles were aligned along parallel fibrils or were found singly in some instances. As incubation continued, the particles became increasingly more spherical, and formed parallel long rows, and (iii) eventually, 24 hr after inoculation, formed closely packed viral crystals of a rectangular or hexagonal lattice. The mature progeny particles were 26 to 28 mμ in diameter. These changes were reminiscent of those reported previously for the polio and other enteroviruses.
Morphological and Biological Changes Induced by Thallium Acetate in Escherichia coli
Authors: Nakamura Masahiro , Kohguchi Kumiko
Published: November 2013

Morphological changes in Escherichia coli C‐600, from an elongated to a spherical form, induced by addition of 500 μg thallium acetate (TAL)/ml of nutrient agar are described. Properties of the TALR E. coli (resistant to TAL) were studied, comparing to those of original E. coli: the TALR E. coli cells were more fragile than those of the original cells in water and 3.5% NaCl solution, and more sensitive to heating, antibiotics except penicillin, and lysozyme. However, there were no differences of cell wall structure observed by an electron microscope and glucosamine contents of cells measured by chemical analysis between TALR E. coli and original one. A few cells of TALR E. coli were elongated by platinum ions, whereas most of the cells of original E. coli were elongated. Multiplications of coliphage T2 in TALr E. coli cells were faster and more numerous than in those of the original ones. Therefore, it might be presumed that morphological changes in E. coli C‐600 induced by thallium acetate would be caused by less rigidity in the cell wall constitution, without any loss of cytoplasmic function.
Experimental Infection with Pseudomonas aeruginosa in Mice
Authors: Kobayashi Fujio
Published: November 2013

Comparative studies on the virulence of 22 clinical isolates of Pseudomonas aeruginosa were made by means of intraperitoneal inoculation in mice. The LD50 values of these strains for mice ranged from 105 0 to 107.5 viable cells per mouse and the average was 106.6. The virulence of certain strains was significantly enhanced when these strains were inoculated mixed with mucin. The highly virulent strains were often found among the strains which were serologically untypable though no relationship could be found between the virulence of test strains and their other biological characteristics such as pigments, hemolysins and extracellular enzymes. The facts suggest that pigments and extracellular enzymes play no important role in the pathogenicity of P. aeruginosa for mice. Moreover, no difference was seen on virulence among the strains isolated from the patients and healthy carriers. The susceptibility of ICR, ddN and CF#1 mice for P. aeruginosa was investigated. There was no clear difference in susceptibilities to P. aeruginosa infection.
Slime Production by Pseudomonas aeruginosa
Authors: Goto Sachiko , Enomoto Seiji , Takahashi Yoshio , Motomatsu Reizo
Published: November 2013

Adequate experimental conditions for slime production by Pseudomonas aeruginosa were investigated using a cellophane plate method. Definite slime production was observed on heart infusion agar, brain heart infusion agar, yeast extract agar and synthetic agar, but not on nutrient agar. The addition of phosphate to the nutrient agar above 0.05% caused visible slime formation. Incubation at 37 C resulted in a higher yield of slime than at 25 C. Longer incubation seemed more favorable for slime production, while the pH reaction of the test media did not effect the slime yield. All the test cultures of P. aeruginosa produced large amounts of slime by this procedure. Cultures of Pseudomonas fluorescens, other Pseudomonas spp. and certain vibrios also produced slime under these experimental conditions.
Studies on the Persistent Infection with Measles Virus in HeLa Cells
Authors: Minagawa Tomonori
Published: November 2013

HeLa/MV cells which have previously been found to contain measles antigen in more than 90% of cells were very similar to normal HeLa cells in their morphology and growth. Although almost all of HeLa/MV cells may be infected, only 10% of them released infectious virus particles. The hemadsorption test, however, showed that most of the infected cells produced by hemagglutinin. Two kinds of clones were obtained by cloning HeLa/MV cells in the presence of anti‐measles serum. One was the virus‐releasing clone and the other the uninfected clone which did not contain any measles antigen. The proportions of virus‐releasing clones to all clones varied between 20 to 54%, and did not show any increase even after recloning of the virus‐releasing clones. The susceptibility of the uninfected clones to the standard measles virus was not different from that of normal HeLa cells.
Studies on the Persistent Infection with Measles Virus in HeLa Cells
Authors: Minagawa Tomonori
Published: November 2013

HeLa cells were caused to degenerate by infection with the standard measles virus, while those infected with the virus released from the HeLa/MV culture (carried virus) readily established a carrier state. The carried virus had a longer latent period and a lower growth rate in both HeLa and Vero cell cultures as compared with the standard virus. Plaques by the carried virus on Vero cell cultures were delayed in appearance as compared with those by the standard virus. However, when the carried virus was passed once through Vero cells, the virus became able to form plaques as readily as the standard virus. Also the passed virus was found to have lost the ability to establish a carrier state. These results may suggest the possibility of a host‐controlled variation, though the mechanism of establishment of the HeLa/MV culture which harbors such a variant remains obscure.
Erythrocytes of Japanese Quail for Hemagglutination by Rubella Virus
Authors: Soda Kenji , Matumoto Minoru , Tabata Eizo
Published: November 2013

Erythrocytes (RBC) of adult Japanese quail, Coturnix coturnix japonica, were found to be as sensitive as day‐old chick RBC for rubella virus hemagglutination (HA). Various factors involved in the HA were studied with quail RBC as well as with adult pigeon and day‐old chick RBC. Pigeon RBC, unlike chick and quail RBC, tended to show, without hemagglutinin, a pattern of sedimented RBC resembling the HA pattern by the virus at 4 C, to a lesser extent at room temperature, in 0.85% NaCl solution buffered with m/100 phosphate (PBS), and were prevented from doing so by addition of a small amount of bovine plasma albumin to the diluent. A small amount (about 10−3 m) of CaCL2 in PBS gave higher HA titers. The HA titer was higher at 4 C than at room temperature and much reduced at 36 C with chick and quail RBC. With pigeon RBC, addition of bovine plasma albumin to the diluent tended to reduce HA titers. The HA titer was highest at pH 5.8 to 6.8, and was inversely proportional to the RBC concentration. Under the conditions established on the basis of these findings, chick and quail RBC gave similar HA titers, but pigeon RBC gave consistently higher titers. However, these types of RBC gave no significant difference in HA‐inhibiting antibody titer when 4 units of hemagglutinin as determined with the homologous RBC was used.
Lipids of Sendai Virus and Changes in Cellular Phospholipid Synthesis Caused by Infection
Authors: Hirano Koichi , Yamaura Koichi , Shibuta Hiroshi , Matumoto Minoru
Published: November 2013

The labeled lipid composition of Sendai virus grown in chick embryo (CE), cynomolgus monkey kidney (MR) and bovine kidney (BK) cells differed from that of these host ceils. In 20 hr labeling after infection with 32P‐orthophosphate, 14C‐acetate or 14C‐palmitic acid, virions had a high content of labeled phosphatidylethanolamine (PE) and a lower content of phosphatidylcholine (PC), while the host cells showed a reverse result. Virions also had a higher proportion of sphingomyelin than the host cells. Similar phospholipid patterns were also obtained by infecting cells labeled with 32P‐orthophosphate prior to infection. CE and MK cells labeled with 32P‐orthophosphate before infection followed by a 20‐hr labeling with 3H‐acetate revealed that lipids containing fatty acids newly synthesized after infection were incorporated preferentially into virions. The cause of these differences in lipid composition between the virion and the host cell should be sought. Relevant to this question was the finding that Sendai virus infection enhances not only total phospholipid synthesis, but also the PE and PC synthesis and presumably the pathway from phosphatidylserine to PE. These metabolic changes induced by infection may affect the lipid composition of the plasma membrane, hence that of the viral envelope.
Studies on the Antigenic Activities of Yeasts
Authors: Suzuki Shigeo , Hatsukaiwa Hisanori , Sunayama Hiroyoshi , Honda Naoko
Published: November 2013

Five antigenic mannans isolated from the cells of Candida albicans serotype A, C. albicans serotype B, C. tropicalis, C. stellatoidea and Saccharomyces cerevisiae were examined for their reactivities against concanavalin A, the size of the combining site has been estimated to be relatively small, up to 4 hexopyranosyl residues. In the quantitative precipitation reaction, all mannan‐concanavalin A systems afforded nearly the same amounts of nitrogen or mannan precipitated, and the ratios of precipitation‐inhibition with α1→2 linked manno‐oligosaccharides, from biose to tetraose, were also equal regardless of the structural differences of these mannans. Furthermore, in agar‐gel double diffusion analysis, all the systems gave a corresponding precipitation arc which completely fused with the adjacent ones. These behaviors of mannan‐concanavalin A systems resemble those of antigen‐antibody systems consisting of the same mannans and anti‐C. albicans serotype B serum. It also provided evidence that the previous interpretation on the lesser serologic specificity of this serum compared to that of anti‐C. albicans serotype A was due to the smaller size of combining sites for antibodies of the former than of the latter serum.
Intestinal Resistance in the Experimental Enteric Infection of Mice with a Mouse Adenovirus
Authors: Hashimoto Kazuo , Yoshikawa Masako , Sugihara Yumiko , Sasaki Shogo
Published: November 2013

In order to find out whether the mouse adenovirus‐neutralizing substance, which appeared in the intestinal tract of mice orally infected with mouse adenovirus, was an immunoglobulin, examinations were carried out for the status of 3 classes of immunoglobulin, IgA, IgG, and IgM, in the intestinal tract as well as in the serum of the mouse. In infected mice, as in uninfected mice, the serum contained much IgG, a moderate amount of IgA, and a small amount of IgM, whereas the intestinal wall showed a moderate amount of IgA, a small amount of IgG and no IgM, and the intestinal contents contained a moderate amount of IgA. Secondly, DEAE‐cellulose chromatography or Sephadex G‐200 gel filtration was done in order to know whether the virus‐neutralizing activity was recoverable in the fractions containing some class of immunoglobulin. The result indicated that a large part of the activity in the serum was recovered in the fractions of IgG and a small part in those of IgA. In the case of the intestinal wall, a large part of the activity was found in the fractions of IgA, and only a small part in the fractions containing both IgG and IgA. In the intestinal contents, the activity was detected solely in the fractions containing IgA. Finally, when the substance from the intestinal wall was purified by DEAE and Sephadex, a parallel increase of both IgA and the virus‐neutralizing activity per protein content was observed. Thus, it became clear that the mouse adenovirus‐neutralizing substance in the intestinal tract was an antibody against the virus, and that it mostly belongs to IgA.
Interferon‐Inducing Activity of Acidic Polysaccharides
Authors: Suzuki Shigeo , Suzuki Masuko , Maya Masanori I , Chaki Fumio
Published: November 2013

Readily available polysaccharides, amylopectin, amylose, dextrin, and yeast mannan, were chemically phosphorylated using polyphosphoric acid in the presence of a tertiary amine, and the resultant phosphates were examined for their interferon‐inducing activity in rabbits employing an assay system consisting of a primary culture of rabbit kidney cells and vesicular stomatitis virus. All the phosphates were shown to be active as interferon inducer, and, especially, the activity of those containing more than 2% phosphorus were quite strong. Interferons evoked by the above phosphates resembled those induced by bacterial endotoxin, e.g., the viral inhibiting activity was susceptible to heat treatment, low pH and tryptic digestion. Since all the parent polysaccharides showed no interferon‐inducing activity, it is reasonable to assume that the active center of these inducers might reside or be due to the anionic phosphate groups.
In Vivo Induction of Resistance to Macrolide Antibiotics in Staphylococci with Erythromycin
Authors: Kurashige Satonori , Inoue Matsuhisa , Yamaguchi Nobuo , Mitsuhashi Susumu
Published: November 2013

In vivo induction of resistance to macrolide (Mac) antibiotics was investigated using mice infected with Staphylococcus aureus MS537 followed by treatment with either erythromycin (EM) or josamycin (JM). An EM‐inducible strain MS537, in which only EM was an active inducer for Mac resistance, acquired in vivo resistance to both Mac antibiotics and lincomycin (LCM) 2 hr after an EM injection but the resistance of the induced cells was lost 2 days thereafter. Consequently, EM was found to have no therapeutic effect on MS537 infection and the number of resistant bacteria recovered from the kidney increased even after 6 intraperitoneal injections of 0.2 nig each of EM; the number of bacteria recovered from the kidney showing the same level as the controls without treatment. JM, one of the non‐inducers for Mac resistance, by contrast, showed a therapeutic effect on MS537 infection and the number of organisms recovered from the kidney decreased to one‐thousandth of that obtained with EM treatment, resulting in the survival of mice.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Mashiba Harukazu , Takeya Kenji
Published: November 2013

Antibody response against hamster red blood cells (H‐RBC) was examined in inbred strains of C57BL/6, AKR, C3H/He, DDD and SL mice, and outbred CF1 mice. 1) There were strain differences in antibody response after a primary intravenous injection of H‐RBC. DDD, SL and CF1 mice belonged to high‐responder strains, while C57BL/6, AKR and C3H/He to low‐responder strains. In the spleens of immunized CF1 and SL, 40 to 70 times as many plaque‐forming cells (PFC) as those in C57BL/6 mice were detected. The magnitudes of the response were: CF1 ≒ SL>DDD>>C3H/He ≑ AKR>C57BL/6. 2) 2‐mercaptoethanol resistant (MER) antibody was detected in neither low‐ nor high‐responders after a primary intravenous antigen‐injection. 3) After a secondary intravenous antigen‐injection, MER antibody was detected in all the SL mice, but only in 30 to 50% of AKR and C57BL/6 mice. 4) A subcutaneous injection of H‐RBC in Freund's complete adjuvant (FCA) did not elicit antibody production within 10 days. When mice pre‐sensitized 7 days in advance wTith H‐RBC in FCA were intravenously injected with H‐RBC, enhanced antibody production of the primary type was observed in all the mouse strains. 5) In pre‐sensitized mice, the extent of the enhancement of antibody production was the highest in low‐responder C57BL/6 mice and the lowest in high‐responder SL and CF1 strains. Thus, there was no strain difference in antibody titers or the numbers of PFC after the booster.
Comparative Studies on Large‐ and Small‐Plaque‐Forming Clones of Newcastle Disease Virus (Miyadera Strain)
Authors: Kato Nobuo , Ohta Fujito , Kato Osamu
Published: November 2013

The Miyadera strain of Newcastle disease virus (NDV) consisted predominantly of virus particles forming small plaques on monolayers of chick embryo fibroblasts (CEF), and contained small amounts of virus particles forming large plaques. These large‐ and small‐plaque‐forming clones of this virus (NDV‐L and NDV‐S) were isolated. The small size of the NDV‐S plaques did not appear to be due to an agar inhibitor. NDV‐L produced a much higher yield of infective virus particles in CEF and they were released more completely from the infected cells than were those produced by NDV‐S. The yield of infective virus of NDV‐L per cell from cultures of CEF was comparable to the yield from the allantoic cells. The infectivity/hemagglutinin ratio for NDV‐L from CEF was as high as the ratio for virus from the allantoic cells, but the ratio for NDV‐S from CEF was lower. NDV‐S demonstrated an autointerference phenomenon in CEF when infected at high multiplicities, but NDV‐L did not. Contrary to virus multiplication, NDV‐S exhibited a more rapid and marked cytopathic effect on monolayers of CEF than NDV‐L. In the allantoic cavity of eggs NDV‐S produced slightly higher virus yields than NDV‐L. No correlation existed between plaque size of the two viruses and the capacity to induce interferon synthesis or the susceptibility to the action of interferon. The properties of both distinctive plaque isolates were stable on egg passage.
Lethal but Non‐Pyrogenic Products of Endotoxin Obtained after Hydrolysis with Lithium Hydroxide
Authors: Konno Seishi , Yoshioka Morimasa
Published: November 2013

An endotoxin was extracted from Salmonella typhosa, strain O‐901, with trichloroacetic acid and phenol‐water (TCA‐PW ET). TCA‐PW ET was treated with 0.5 N lithium hydroxide at 37 C for 30 min. The hydrolysate was chromatographed in a Sephadex G‐150 column. The absorbancy at 485 mμ showed five successive peaks (Frs. 1 through 5). Analytical centrifugation indicated that these fractions were composed of diverse sizes. The sedimentation coefficient of Fr. 5 was the smallest being approximately 6.5 S. The composition of each fraction was shown to be different by chemical analysis. Fraction 5 consisted mostly of hexosamine and fatty acids and some neutral sugars. With gas chromatography Fr. 5 showed a distinct difference when compared with either TCA‐PW ET or Frs. 1 to 3 in its reduced lauric acid as well as increased β‐hydroxymyristic acid contents. Rabbit pyrogenicity of Fr. 5 as compared with TCA‐PW ET was approximately one thousandth, while chick embryo lethality was almost the same. Biological activities of the other fractions were intermediate in relative potencies. Fr. 5 was further chromatographed in Sephadex G‐75 and G‐50 columns and six fractions were obtained. All of these fractions were highly lethal with slight differences. Phosphorus, heptose, 2‐keto‐3‐deoxyoctonate and glucosamine contents of the endotoxin products were relatively high, while the fatty acid contents were partly decreased. Relationship between lethality and chemical constituents of the endotoxin products is discussed.
Effect of the Live and Heat‐Killed Cells of Staphylococcus aureus on the Resistance of Mice to the Infection with Nocardia asteroides
Authors: Uesaka Ichiro , Kobayashi Hiroshi
Published: November 2013

When living cells of Staphylococcus aureus were introduced into mice by various routes within a few hours before or after infection with Nocardia asteroides, a marked increase in mortality and enhanced severity of lesions were observed. This effect was presumably caused by a heat‐labile substance(s) liberated from the cells of S. aureus. No increase in resistance occurred when S. aureus was given 2 to 7 days before the nocardial infection, contrary to the effect of E. coli as reported previously. An increased resistance was noted when heat‐killed cells of E. coli were administered two to four days before concurrent infection with N. asteroides and S. aureus.
Electron Microscopic Studies on the Replication of Ibaraki Virus
Authors: Saito Yasuji , Suzuki Yujiro , Nakagawa Susumu
Published: November 2013

Ibaraki virus is an agent of epizootic cattle disease resembling bluetongue. The virus was purified from infected BHK‐21 cells and bovine kidney cells. The particle negatively stained was icosahedral and had a diameter of 45–50 μ. By electron microscopy, the virus was found in the cytoplasm of cells, assembling in lysosomal or matrix‐like dense bodies. In early stages of infection, 2 to 7 hr, virus particles were located in vesicles, simulating a viropexis, whereas at 24 to 30 hr, after infection, a large number of cores of progeny virus were scattered in matrix‐like bodies or packed and crystallized in lysosomal bodies bounded by a membrane. Sometimes peculiar tubular structures were found near the matrix‐like bodies. The relationship between these structures and virus replication was not clear. The morphological features of the particles and virus development in cells resembled those of viruses of bluetongue, African horse‐sickness, Colorado tick fever and reo.
Cell‐Cooperation in Anti‐Sheep Red Blood Cell Antibody Response in Mouse Spleen Cell Cultures
Authors: Tadakuma Takushi
Published: November 2013

Restoration of impaired antibody response to sheep red blood cells (SRBC) in spleen cell cultures from mice treated with heterologous antilymphocyte globulin (ALG) was studied by adding normal cells from various sources, to explore the problems of cell‐cooperation in anti‐SRBC antibody response and the target of ALG. When spleen cells from ALG‐treated mice were separated into macrophage‐rich and lymphoid cell‐rich subpopulations, only the latter was found to be impaired in the ability for anti‐SRBC antibody response. Addition of even a small number of normal allogeneic spleen cells sufficiently restored the impaired anti‐SRBC antibody response of the spleen cells from ALG‐treated mice. By use of allo‐antisera, most hemolysin plaque‐forming cells (PFC) generated in such cultures were proved to be derived from the cells of ALG‐treated mice. Restoration was also achieved by adding thymus‐derived cells, which were obtained from spleens of mice heavily irradiated and repopulated with syngeneic thymus cells, or lymphoid cells directly collected from thymuses. All results indicate that ALG selectively depletes the thymus‐derived antigen reactive cells (ARC) in the spleen cell population, and that ARC supplied from normal spleen or thymus can interact with plaque‐forming cell precursors (PFCP) that remain intact in the spleen cell population of ALG‐treated mice. The results also suggest that a single ARC interacts with more than one PFCP and makes them develop into PFC.