bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of Immunoglobulin Production from Human B Lymphocytes by Staphylococcus aureus: Effects of Monocytes and Con A‐Induced Suppressor Cells
Authors: Kasahara Tadashi , Harada Hirotomo , Ogata Kenji , Kawai Tadashi , Shioiri‐Nakano Kohei
Published: November 2013

Significant immunoglobulin (Ig) production by human peripheral blood lymphocytes was induced in vitro by stimulating the cells with pokeweed mitogen (PWM) and Staphylococcus aureus Cowan I (SpA CoI). IgG, IgM, and IgA were determined by a combination of the latex fixation test and radioimmunoassay. High levels (1,000 to 5,000 μg/ml) of IgG and IgM and a lesser amount of IgA were constantly produced during 7 to 8 days of incubation with both stimulants. Ig production induced by SpA Col stimulation was independent of the presence of T cells, while Ig production induced by PWM required T cells exclusively. Depletion of monocytes in the culture caused but a slight decrease in Ig production (particularly in the case of IgG). While the addition of a small number of monocytes enhanced IgG induction by both stimulants, coculture with an excess number of monocytes inhibited Ig induction (particularly IgG) by PWM stimulation but not by SpA CoI stimulation. Marked suppression of Ig production (IgG, IgM, and IgA) was observed in cocultures with Con A‐activated T cells. The phenomena of suppression were observed in both the SpA Col‐stimulated and PWM‐stimulated lymphocytes. These data indicate that Ig production from B cells stimulated with a polyclonal B cell activator, SpA CoI, was independent of T cells and relatively of independent of monocytes, but could be subjected to the regulation of the Con A‐induced suppressor T cells.
Comparison of Ciliostasis by Mycoplasmas in Mouse and Chicken Tracheal Organ Cultures
Authors: Araake Minako
Published: November 2013

The cilia‐stopping effect of mycoplasmas of human and various animal origin in mouse and chicken tracheal organ cultures was studied. From the results in mouse tracheal organ cultures, the mycoplasma strains tested were divided into three groups: Mycoplasma pulmonis m53, M. pulmonis JB, M. pulmonis OK, M. mycoides subsp. mycoides PG1 and M. gallisepticum S6 showed a strong cilia‐stopping effect; M. pulmonis PG22, M. mycoides subsp. capri PG3, M. meleagridis 19729, M. neurolyticum Type A and M. arthritidis PG6 showed a mild effect; and M. pneumoniae FH, M. salivarium Hup, M. hominis type 1‐C and M. orale N‐C of human origin and Acholeplasma laidlawii PG8 showed a weak effect. On the other hand, in chicken tracheal organ cultures, only M. gallisepticum S6 showed a strong effect, M. meleagridis 19729 was affected to a lesser degree, and other mycoplasma strains showed a weak or no effect. The results indicate that some murine and poultry mycoplasmas showed a cilia‐stopping tendency in mouse and chicken tracheal organ cultures, respectively, while human mycoplasmas showed weak or little effect in both organ cultures. In mouse tracheal organ cultures, M. pulmonis m53 treated with heat, trypsin or formaldehyde, and the sterile filtrate of an m53 broth culture showed no cilia‐stopping effect. The relationship of the pathogenicity of mycoplasmas for their natural hosts to that for cultured respiratory cells is discussed.
Properties of T Antigen‐Positive Phenotypic Revertants Isolated from SV40‐Transformed Rat and Mouse Cells
Authors: Maruyama Kazuo , Furuta Takahisa , Ueda Katsumoto , Oda Kinichiro
Published: November 2013

Four T antigen‐positive phenotypic revertants were isolated by negative selection with BUdR from SV40‐transformed rat and mouse cells which contain six and two viral genome equivalents per cell, respectively. Karyological analysis indicated that one rat and one mouse revertant had a hyperploid number of chromosomes, while the remaining two rat revertants had a subtetraploid number similar to those of the transformed parent cells. The hyperploid revertants were unable to grow in soft agar medium and were nontumorigenic in nude mice. One of the subtetraploid revertants formed large colonies at a very low frequency and induced tumors after a prolonged incubation period. These results indicate that there is a good correlation between the capacity of cells to grow without anchorage and the capacity to form tumors in nude mice and suggest that the revertant phenotype is stable in the presence of T antigen when the number of chromosomes is greatly increased as compared with that of the transformed parent cells.
Nonbacteremic Pseudomonas Pneumonia in Immunosuppressed Guinea Pigs
Authors: Otani Tsuyoshi , Katami Kazuei , Une Tsutomu , Osada Yasuaki , Ogawa Hidemasa
Published: November 2013

An experimental model of nonbacteremic pneumonia with a virulent strain of Pseudomonas aeruginosa was successfully established in guinea pigs immunosuppressed with cortisone acetate although the organisms were eliminated rapidly from the lungs without cortisone treatment. Using a pocket nebulizer, almost all the animals which received 106 organisms/g‐lung developed bronchopneumonia without any septic findings as long as 10 days after challenge. The lesions produced in such animals were characterized by dissemination of multiple purulogranulomatous changes. In the early stage of infection, infiltration of polymorphonuclear cells (PMNs) in the bronchiolar and alveolar spaces was diffuse, later showing multifocal accumulation with the formation of central spherical grains enclosing bacterial colonies. In the later stage, granulation tissue consisting of large mononuclear cells, fibroblasts and collagen fibers developed around the PMN accumulation. The animals which received 107 organisms/g‐lung, on the other hand, developed severe pulmonary hemorrhages and necrosis followed by septic death.
Flagellation of Pseudomonas aeruginosa during the Cell Division Cycle
Authors: Amako Kazunobu , Umeda Akiko
Published: November 2013

Flagellation of Pseudomonas aeruginosa during the cell division cycle was examined by scanning electron microscopy. A new flagellum grows on an old polar end located at the opposite position of the parental flagellum in the late stage of the cell cycle.
Effect of Histamine‐Sensitizing Factor of Bordetella pertussis on Pharmacologic Response of Rat Atria
Authors: Endoh Masahiko , Nakase Yasukiyo
Published: November 2013

The effects of sensitization with the histamine‐sensitizing factor (HSF) of Bordetella pertussis as well as Bordetella vaccines on a pharmacologic response in rat heart preparations were determined. In normal rats the spontaneous beating of atria in vitro through the positive inotropic action produced by the addition of epinephrine was inhibited immediately by addition of acetylcholine, whereas in the B. pertussis vaccine‐treated rats the exciting atria were scarcely inhibited by acetylcholine. Neither B. parapertussis nor B. bronchiseptica vaccines induced such an altered atrial response in rats. Of the B. pertussis cell components purified HSF induced the altered response at the minimal dose of 0.1 μg per rat, and a dose of 1 μg or more produced the maximal change. This altered atrial‐inducing activity of HSF was inactivated by heating at 63 C for 30 min, and was neutralized by anti‐HSF rabbit serum. The altered response rose quickly in 1 day after i.v. injection of 1 μg of HSF, reached a plateau in 3 to 5 days, which lasted at least 14 days, and disappeared completely 56 days later. HSF failed to produce directly any functional damage to the beating atria in vitro, and to induce the altered response of the normal rat atria by incubation with as much as 10 μg of HSF per bath (50 ml) for 4 hr. A trace stimulation was found in the normal rat atria as well as in perfused frog hearts, if HSF was given directly at a dose of 20 μg per bath.

The possible mechanism of the altered atrial response in the HSF‐sensitized rats is discussed.
Alterations in the Immunogenic Properties of Sheep Erythrocytes by Sonic Disruption
Authors: Nagase Fumihiko , Nakashima Izumi , Nagase Noriko , Kato Nobuo
Published: November 2013

A solubilized sheep red blood cell (SRBC) antigen (supernatant fraction obtained by centrifuging 107‐2 × 108 sonicated SRBC at 6 × 104 g for 30 min [Sup‐SRBC]), whose ability to inhibit anti‐SRBC plaque formation was 70% of that of the original sonicated SRBC, was unable to elicit a detectable antibody response in either unprimed or SRBC‐primed mice. However, Sup‐SRBC as well as intact SRBC antigens generated memory for the secondary response, which was transferable to irradiated syngeneic recipients by injection of immune spleen cells. The memory generated by Sup‐SRBC involved helper memory for anti‐trinitrophenyl group (TNP) response to challenge with TNP‐conjugated SRBC. Increase in the helper T cell memory in the spleens of Sup‐SRBC‐primed mice was also demonstrated by an in vitro culture experiment and by an adoptive cell transfer experiment. In contrast, no detectable B cell memory was generated by Sup‐SRBC. Repeated stimulation with Sup‐SRBC never induced significant antibody response but reduced the level of memory. A single injection of a low dose (106) of SRBC also failed to induce a definite primary antibody response generating memory for the secondary response. However, repeated stimulation with this dose of SRBC induced a high antibody response and generated good memory. From these results it is suggested that the intact structure of SRBC is required for the activation of B cells, but is not necessary for the stimulation of T cells.
Properties of Hg‐ and Cd‐Spores of Bacillus megaterium QM B1551
Authors: Imagawa Masayoshi , Ichikawa Tomio , Kondo Masaomi
Published: November 2013

Hg‐ and Cd‐spores of Bacillus megaterium QM B1551 were produced in Schaeffer's medium containing mercuric chloride and cadmium chloride respectively. Metals were added to the medium at 9 hr of incubation (Stage V) to give a final concentration of 50 μm. It was found by electron microscopic and biochemical studies that the coats of both Hg‐ and Cd‐spores were thinner than those of control spores. Of the total Hg and Cd in the spores, 77% of the Hg and 63% of the Cd were detected in the spore coats.

Hg‐ and Cd‐spores were less resistant to heat and more sensitive to germinants than control spores. Other properties of Hg‐ and Cd‐spores were similar to those of control spores. These results suggest that the spore coat has some relationship to the heat resistance and germinability of spores.
Regulatory Factors of Lymphocyte‐Lymphocyte Interaction: II. Characterization of Human Mitogenic Factor Derived from the Culture Supernatant of a Mouse‐Human Hybridoma
Authors: Kawano Michio , Namba Yuziro , Hanaoka Masao
Published: November 2013

Cell hybridization of phytohemagglutinin (PHA)‐stimulated human peripheral blood lymphocytes (PBL) with murine lymphoma (EL‐4) provided three hybridomas (MHH‐16, MHH‐20, and MHH‐22) which spontaneously produced human mitogenic factor (MF). MHH‐16 was serially subcloned by limiting dilution procedures, which resulted in maintaining two subclones producing human MF spontaneously for more than one year (PQL‐3 and PQL‐5 subcloned lines). Human MF (MHH‐MF) derived from supernatants of PQL‐5 line cultures had a molecular weight (m.w.) of about 26,000–30,000 daltons (the major peak) with a minor peak with an m.w. of 15,000 daltons on Sephadex G‐100 chromatography, and at a high concentration of NaCl (1 m), the activity of the 26,000–30,000‐m.w. fraction became weak and that of the 15,000‐m.w. fraction became predominant. MHH‐MF had an isoelectric point of pH 5.0–6.5. On DEAE‐cellulose chromatography, MHH‐MF was eluted at a fairly low salt concentration (sodium phosphate buffer 0.02 M, pH 8.0, NaCl 10 mm). After periodate treatment of this MHH‐MF, the mitogenic activity almost disappeared. MHH‐MF was relatively unstable to heating at 56 C for 20 min. In the presence of tunicamycin (0.3μg/ml), an inhibitor of N‐linked glycosylation, the synthesized MHH‐MF showed a decrease in m.w. as follows: the major peak shifted from 26,000–30,000 to 23,000 daltons and the minor peak from 15,000 to 10,000 daltons on Sephadex G‐100 chromatography.

In internal labeling experiments with [3H]leucine, the 3H‐labeled MF was partially purified, with mitogenic activity as a guide. This 3H‐labeled MHH‐MF fraction could be absorbed by PHA blasts but not by normal PBL. On SDS‐PAGE under reducing conditions, only the radioactive peak of the 15,000‐dalton fraction was recovered. MHH‐MF obtained from the hybridoma culture supernatants may be a dimer of the 15,000‐dalton fraction and a glycoprotein.
Isolation and Properties of Dextranases from Bacteroides oralis Ig4a
Authors: Takahashi Nobuyoshi
Published: November 2013

Extracellular dextranases were extracted from a dextran‐degrading microorganism, Bacteroides oralis Ig4a, which had been isolated from human dental plaque, and purified.

Crude enzyme preparations obtained from a broth culture supernatant by salting out with ammonium sulfate were subjected to column chromatography on DEAE‐cellulose and subsequent Bio‐Gel p‐100, followed by isoelectric focusing.

Two kinds of enzyme preparations, Enzymes I and II, with the ability to degrade soluble dextran were obtained. The optimal pHs of Enzymes I and II were 5.5 and 6.8, and the isoelectric points were pH 4.5 and 6.5, respectively. The molecular weights of Enzymes I and II were estimated by SDS‐PAGE to be 44,000 and 52,000. Both enzymes were inhibited by Pb2+ and Fe3+, but not by Ca2+, Mg2+, Zn2+, or Fe2+. Neither the presence of EDTA nor iodoacetamide had any appreciable effect on the enzyme activity. The enzyme activity was independent of any of these metal ions.

Enzyme I liberated glucose, isomaltose, maltotriose and higher oligosaccharides from dextran. In contrast, Enzyme II liberated only glucose from dextran and was assumed to be an exoglycosidase.

Neither of the enzymes degraded modified insoluble glucan, which is a partially oxidized mutan of S. mutans containing predominantly α‐(1, 3) linkages.
Glycerol‐Releasing Activity of Histamine‐Sensitizing Factor of Bordetella pertussis for Rat Adipocytes In Vitro
Authors: Endoh Masahiko , Nakase Yasukiyo
Published: November 2013

Histamine‐sensitizing factor (HSF) purified from Bordetella pertussis induced specifically the release of glycerol from rat epididymal adipocytes in vitro. The most sensitive and reproducible results were obtained by using 1 to 2 × 105 adipocytes/tube from rats weighing 150 to 200 g, and by incubation at 37 C for 180 min. After a lag period of about 60 min, HSF‐treated adipocytes released glycerol in increasing amounts between 60 and 240 min, depending on the dose of HSF. A close correlation between the glycerol‐releasing (GR) activity of HSF for adipocytes and histamine‐sensitizing or leukocytosis‐promoting activity in mice was observed. The GR activity was inactivated by heating at 56 C for 60 min, 63 C for 30 min or 96 C for 10 min. The adipocytes washed out with a Krebs‐Ringer bicarbonate buffer immediately after being exposed to HSF for 1 to 3 min manifested about 75% of the total GR activity induced by HSF, and those washed out after being exposed for 30 min or longer had full activity. Anti‐HSF serum neutralized the activity when it was added to adipocytes simultaneously with HSF, but did not when it was added 30 min after being exposed to HSF. By using both native and 125I‐labeled HSF, the ratio of binding of HSF to adipocytes was estimated to be 10 to 15% of the total HSF per 2 × 105 cells/tube, and to be about 1,000 molecules of HSF per cell to induce the release of glycerol. The GR activity induced with 10 ng of HSF was inhibited by addition of insulin at a dose of over 1 μIU/tube, but not by concanavalin A.
A Simple Bile‐Disk Method for the Identification of Bacteroides fragilis
Authors: Yamazaki Etsuko , Sugimoto Kiyoshi , Niwano Kiyoshi , Okada Jun
Published: November 2013

A simple method for the separation and identification of Bacteroides fragilis from other Bacteroides species through the use of oxgall‐impregnated filter paper disks is described. As in the antibiotic sensitivity test, filter paper disks containing defined amounts of bile salt were placed on a lawn of bacterial cells on Gifu anaerobic medium agar plates. Bile‐resistant bacteria were identified as strains of Bacteroides fragilis after incubation in anaerobic jars containing hydrogen and carbon dioxide generators at 35 C for 24 hr. The optimum concentration of bile salt, yielding results very similar to those of the conventional tube method, was determined to be 25 mg per disk, empirically. Since this method is easy to perform, time saving, economical and gives clear results, it may be readily used as a daily routine test in the clinical microbiology laboratory.
R483 and F Plasmid Genes Promoting RNA Degradation: Comparative Restriction Mapping
Authors: Akimoto Shigeru , Ohnishi Yoshinari
Published: November 2013

The gene promoting nucleic‐acid degradation (pnd) on IncIa plasmid R483 was cloned into pBR322. It is located on a 0.85 kilobase (kb) EcoRI‐SalI fragment and is close to Tn7. The pnd gene has similar properties to the srnB gene on the F plasmid. A cleavage map of the 0.85 kb pnd fragment was constructed and compared with that of the 1.18 kb EcoRI‐BamHI fragment containing the srnB gene. These two regions showed marked heterogeneity as evidenced by their distinctly different restriction maps. This result suggests separate paths of evolution of the two genes for stable RNA degradation.
Integration of Bovine Leukemia Virus DNA in the Genomes of Bovine Lymphosarcoma Cells
Authors: Onuma Misao , Sagata Noriyuki , Okada Kosuke , Ogawa Yasuki , Ikawa Yoji , Oshima Kanichi
Published: November 2013

Integration of bovine leukemia virus (BLV) in the genomes of infected cells was investigated in cattle with enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL). Southern blot hybridization of BLV cDNA to Eco RI and Xba I restriction fragments of EBL tumor DNAs revealed that: 1) one to four or more copies of proviral DNA were integrated per genome; 2) the restriction pattern of the integrated proviral DNA was the same in two or three different tumors from the same animals; and 3) different patterns were observed among tumors from four different animals. These findings suggest the monoclonal origin of different tumors in an individual animal and the existence of multiple chromosomal integration sites of BLV provirus. DNAs from several SBL tumors were also analyzed with the same restriction enzymes, but with both representative and cDNA3′‐enriched's of BLV RNA. No hybridization bands reactive with representative BLV cDNA could be detected, while several bands appeared to hybridize with cDNA3′‐enriched.
Relationship between Serotype and Certain Biological and Biochemical Characteristics of Strains of the Mycobacterium avium and Mycobacterium intracellular Complex
Authors: Tsukamura Michio , Mizuno Shoji , Murata Hiroshi , Nemoto Hisashi , Yugi Hiroyuki
Published: November 2013

Some relationships among Schaefer's serotypes and biological and biochemical characteristics were observed in strains of the Mycobacterium avium‐Mycobacterium intracellulare complex.

Strains belonging to serotypes 2 and 16 lacked the capacity to utilize n‐ and iso‐butanols as the sole source of carbon in the presence of ammoniacal nitrogen. However, strains of serotype 2 grew at 45 C and lacked arylsulfatase activity (after 14 days), whereas strains of serotype 16 failed to grow at 45 C and showed positive arylsulfatase activity. Strains belonging to serotypes 8, 9, and 15 grew at 45 C, utilized n‐ and iso‐butanols, and showed arylsulfatase activity.
Mechanisms of Resistance in Escherichia coli to the Sideromycin Antibiotic No. 216: Isolation and Characterization of the Resistant Mutants
Authors: Miyakawa Satohide , Noto Takao , Okazaki Hiroshi
Published: November 2013

Escherichia coli easily developed resistance to a new antimicrobial agent of the sideromycin group, No. 216, by spontaneous mutation. Most of the No. 216‐resistant mutants tested proved not to be cross‐resistant to E. coli phages T1, T5, and φ80. On the other hand, these phage‐resistant mutants were cross resistant to No. 216. The initial site for binding of No. 216 to the sensitive cells was located, at the ton A gene product (Ton A‐protein) of the outer membrane. However, unlike the phage‐resistant mutants, ton A protein (78K‐protein) in most No. 216‐resistant mutants was intact and these mutants were possessed a particular 87K protein in the outer membrane. It is suggested that No. 216 is taken up by ton A protein and then penetrates into the cell by way of a particular transport system and that a highly mutable portion may exist in this reaction system.
pH‐Dependent Hemolysis and Cell Fusion of Rhabdoviruses
Authors: Mannen Kazuaki , Ohuchi Masanobu , Mifune Kumato
Published: November 2013

Human erythrocytes pretreated with fungal semialkali protease or trypsin became susceptible to hemagglutination by vesicular stomatitis virus (VSV) and rabies virus. Both viruses exhibited extensive hemolytic and fusion activities against erythrocytes pretreated with these enzymes. The hemolysis and fusion were pH dependent and the activities were most apparent at pH 5.0 and decreased with increase in pH. However, VSV still exhibited slight hemolytic activity at neutral pH. Hemolysis was also dependent on the dose of virus and was inhibited by treatment of the viruses with antiviral antibody. Results of sodium dodecyl sulfate polyacrylamide gel electrophoresis of erythrocyte membranes suggested that most of the carbohydrates were removed from the membrane proteins by the treatment with proteolytic enzymes.
Prevalence Rate and Age of Acquisition of Antibodies against JC Virus and BK Virus in Human Sera
Authors: Taguchi Fumiaki , Kajioka Jitsuo , Miyamura Tatsuo
Published: November 2013

A total of 480 serum samples from donors including 384 children up to 10 years of age were examined by the hemagglutination‐inhibition (HI) test for the rates of prevalence and age of acquisition of HI antibodies against JC virus and BK virus. Among 136 serum samples from various age groups, there were five (4%) with no detectable antibodies against BK or JC virus, 75 (55%) with antibodies against both viruses, 41 (30.1%) with antibodies against only BK virus and 26 (19%) with antibodies against only JC virus. The prevalence of antibodies against JC and BK viruses was 70.5% and 80.8%, respectively, and the mean HI titers (4 × 2n, n≥1) were 4.90 and 4.30. About 50% of the children had acquired antibodies against BK virus by 3 years of age and against JC virus by 6 years of age. These results indicate that dual latent infections with both viruses are common, although independent infections with either virus are predominant in the human population.
The Role of Humoral and Cellular Mediators in Enhanced Mammary Inflammatory Reactions to Staphylococcal Infection in Systemically Immunized Ewes
Authors: Colditz Ian G. , Watson Dennis L.
Published: November 2013

Prior systemic immunization with live Staphylococcus aureus vaccine enhances the early recruitment of neutrophils into nonlactating mammary glands infected with staphylococci. The study investigates the role of humoral and cellular mediators in this phenomenon. Intramammary infusion of bacteria suspended in immune sheep serum did not enhance the inflammatory response to infection in nonimmunized ewes despite the presence of complement in the infused serum. Infusion of complement activated by incubation with zymosan evoked a massive neutrophil influx into mammary secretions by 4 hr after infusion. Hemolytic complement activity was not detected in mammary secretions of immunized or nonimmunized ewes. These findings indicate that, despite the inflammatory effect of complement activation, humoral immune factors did not promote neutrophil migration into infected glands. Mammary glands of systemically immunized ewes stimulated 5 days previously with staphylococcal soluble antigens (SSA) supported larger neutrophil influxes during staphylococcal infection than contralateral glands stimulated with endotoxin 5 days prior to infection. Exudates of SSA‐stimulated glands had significantly higher cell concentrations, prior to infection, than endotoxin‐stimulated glands; however elevated cell concentrations in endotoxin‐stimulated glands of nonimmune ewes did not support enhanced inflammatory responses. These findings suggest that qualitative but not quantitative characteristics of mammary leucocytes influence the inflammatory response to infection in systemically immunized ewes.
Distribution of RNA Coliphages in Senegal, Ghana, and Madagascar
Authors: Furuse Kohsuke , Sakurai Toshizo , Inokuchi Yoshio , Inoko Hidetoshi , Ando Asako , Watanabe Itaru
Published: November 2013

The distribution patterns of RNA coliphages (phages) in Senegal, Ghana, and Madagascar were investigated by collecting sewage samples from domestic drainage in November, 1980. In Senegal, among 65 sewage samples collected mainly from Dakar and its vicinity, 14 (22%) contained RNA phages (16 strains). By serological analysis, 13 of the 16 strains were found to belong to group III. This is consistent with the distribution pattern of RNA coliphages in tropical and subtropical regions of Asia. In Ghana, however, among 106 samples collected from Accra, Suhum, and their vicinities, only seven (7%) contained RNA phages (seven strains) (groups I, II, and III [1:3: 3]). In Madagascar, among 124 samples collected from Antananarivo, Moramanga, and their vicinities, seven (6%) contained RNA phages (seven strains) (groups I, II, III, and IV [1:1:1:4]). In spite of the low isolation frequency, it can be said that Madagascar appears to have a unique distribution pattern (abundance of group IV phages) which differs from that of any other countries we have examined. The generality of the distribution pattern of RNA phages in the tropical region (abundance of group III phages) was thus verified at least in Senegal.
Inhibition of Macrophage DNA Synthesis by Immunomodulators
Authors: Nagao Shigeki , Tanaka Atsushi
Published: November 2013

Oil‐induced guinea pig peritoneal exudate macrophages were found to incorporate actively [3H]thymidine without any tissue fluids such as conditioned medium, lymphokines or inflammatory tissue exudates. The [3H]thymidine incorporation was markedly suppressed by macrophage stimulants such as muramyl dipeptide (MDP) or bacterial lipopolysaccharide (LPS), while glucosamine incorporation was simultaneously increased by these stimulants. The degree of suppression of thymidine incorporation depended on the cell density, the concentrations of the stimulants, and sera or culture media used. The exposure of macrophages to MDP for 30 min was sufficient to cause significant suppression.
Plaque Formation by Tyzzer's Organism in Primary Monolayer Culture of Mouse Hepatocytes
Authors: Kawamura Seiji , Taguchi Fumihiro , Fujiwara Kosaku
Published: November 2013

Tyzzer's disease organism propagated on primary monolayer cultures of mouse hepatocytes and produced definite plaques. In phase contrast microscopy, the organisms were motile in the plaques. Plaque formation was inhibited by antiserum. After serial plaque cloning the organisms still had virulence in mice. To establish a standard plaquing procedure, factors affecting plaque formation were studied. The critical factors in plaque formation were the culture period of host cells before inoculation, medium for suspending the organisms, and temperature and time of infection. A 24 to 36 hr‐preculture of host cells and trypticase soy broth (BBL) as the suspending medium gave the best results. The optimal conditions for infection were 37 C for 90 min. The plaquing efficiency was higher when a larger volume of inoculum was applied to host cell monolayers, suggesting that the organism played an active role in the initial stage of infection.
Production and Purification of Human MG63 Cell Interferon
Authors: Watanabe Yoshihiko
Published: November 2013

MG63 cells induced by Sendai virus were a convenient source of human interferon, which was of the β type in antigenicity. When analyzed by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE), the interferon migrated as a single component with a molecular weight of 22,000 daltons. It was purified to apparent homogeneity by chromatography on Blue Sepharose, followed by SDS‐PAGE, and estimated to have a specific activity of 4 × 108 international units/mg of protein.
Changes in the Intestinal Microflora in Association with Colon Tumorigenesis in Rats Treated with 1,2‐Dimethylhydrazine Hydrochloride
Authors: Morishita Yoshiyuki , Tsukada Taro
Published: November 2013

The present study was undertaken to determine whether the intestinal microflora change during the tumorigenic process in the colon of rats treated with 1,2‐dimethylhydrazone (DMH), and to compare the intestinal microflora of rats with colon tumors induced by DMH with that of rats with gastric tumors induced by N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG). When compared with those in the control animals, the numbers of streptococci and bacteroidaceae were moderately increased in the intestinal tract of DMH‐treated rats before the development of visible intestinal tumors. The DMH‐treated rats bearing small intestinal and colonic tumors were found to have markedly increased numbers of enterobacteriaceae, Clostridium perfringens, streptococci, bacteroidaceae, and bifidobacteria. In DMH‐induction the overgrowth of enterobacteriaceae and/or C. perfringens was found to correlate with the size and number of tumors in both the small intestine and colon. The increased number of streptococci in the DMH‐treated rats was principally due to an increase in the number of the streptococci which did not reduce triphenyltetrazolium chloride (TTC). On the other hand, in the rats with gastric tumors induced by MNNG the numbers of enterobacteriaceae and TTC‐reducing streptococci were remarkably increased in the intestinal tract of only the debilitated animals, and Pseudomonas aeruginosa was detected in all of them. The number of anaerobic gram‐positive cocci was significantly but not remarkably increased in the gastric tumor‐bearing rats compared with the controls. These results indicate that the intestinal microflora of rats may change depending on the gastrointestinal site where tumors develop and the degree of malignancy in tumorigenesis.
Characterization of Canine Distemper Viruses Adapted to Neural Cells and Their Neurovirulence in Mice
Authors: Yoshikawa Yasuhiro , Yamanouchi Kazuya , Morikawa Yuko , Sakaguchi Masahiro
Published: November 2013

Interaction of the Onderstepoort strain of canine distemper virus (CDV) with three established human neural cells, i.e. IMR‐32 neuroblastoma, 118‐MGC glioma and KG‐1 oligodendroglioma, was examined, and adaptation of CDV to these cells was also attempted. The unadapted virus was found to grow at relatively low titers in the three neural cells inducing moderate to minimal cytopathic effects (CPE). The virus was successfully grown at high titers in these cells after 8 to 10 passages. Biological characteristics such as growth rate, morphology of CPE and plaque size changed after adaptation. Analysis by SDS‐polyacrylamide gel electrophoresis, however, failed to show any difference in the molecular weight of component proteins among the unadapted and three adapted viruses.

Inbred DDD strain of mice developed clinical signs after intracerebral inoculation with the unadapted virus but most of them survived with histological lesions of encephalitis. Neuroblastoma‐adapted virus induced only transient clinical signs in some animals with mild encephalitic lesions in the gray matter. Increases in neurovirulence were found for viruses adapted to glioma and oligodendroglioma cells. Almost all mice inoculated with these two viruses at 3 weeks of age died within 8 days with histological lesions consisting of hyperemia, edema, severe degeneration of nerve cells and a few giant cells. Demyelinating lesions in the absence of inflammatory changes were observed in the cerebellum, pons and medulla oblongata of animals inoculated with oligodendroglioma‐adapted virus.
Macrophage Activation by Lactobacillus casei in Mice
Authors: Kato Ikuo , Yokokura Teruo , Mutai Masahiko
Published: November 2013

Effects of Lactobacillus casei YIT 9018 (LC 9018), which has antitumor activities against allogeneic and syngeneic murine tumors, on macrophage functions were examined.

By intraperitoneal (i.p.) injection of LC 9018, acid phosphatase activity and phagocytic activity of peritoneal macrophages were enhanced significantly compared with those of normal peritoneal macrophages. The phagocytic activities showed peaks 2–3 days after the LC 9018‐injection. LC 9018 accelerated the phagocytic function of the reticuloendotherial system in ICR mice tested by the carbon clearance test. The cytostatic activity of peritoneal exudate cells (PEC) induced by i.p. injection of LC 9018 into C57BL/6 mice against EL4 cells was also enhanced. On the other hand, PEC induced by L. fermentum YIT 0159, which has no antitumor activity, did not have cytostatic activity. These observations showed that LC 9018 was able to activate macrophages in mice.
The Klebsiella O3 Lipopolysaccharide Isolated from Culture Fluid
Authors: Hasegawa Takaaki , Ohta Michio , Mori Masashi , Nakashima Izumi , Kato Nobuo
Published: November 2013

In a series of our earlier studies, the O3 antigen isolated from culture supernatant of Klebsiella pneumoniae strain Kasuya (O3:K1) (KO3) was shown to exhibit very strong adjuvant activity in mice. KO3 obtained was homogeneous in analyses by either gel filtration or ultracentrifugation. Its molecular weight determined by ultracentrifugal analysis was greater than 2 × 106. It contained 37.9% C, 6.20% H, 0.24% N, and less than 0.1% P. KO3 was degraded into the polysaccharide moiety and lipid moiety (about 20%) by hydrolysis with 1% acetic acid at 100 C for 1 hr. The molecular weight of the polysaccharide moiety obtained by the hydrolysis was 16,200 as determined by the Somogyi‐Nelson method. Chemical analyses using methylation analysis and Smith degradation as the principal methods indicated that the polysaccharide moiety consisted of a mannan which has a pentasaccharide repeating unit of α‐mannosyl‐1,3‐α‐mannosyl‐1,2‐α‐mannosyl‐1,2‐α‐mannosyl‐1,2‐α‐mannose joined through α‐1,3‐mannosyl linkages. The number of repetitions was less than 20. The fact that minor components such as 2‐keto‐3‐deoxyoctonate and glucose were detected suggests the presence of a core oligosaccharide, but its precise structure is unknown.
Effect of Disruption by Sonication under Different Conditions on the Activity of Glucose Dehydrogenase from Resting Spores of Bacillus subtilis
Authors: Tochikubo Kunio , Yasuda Yoko
Published: November 2013

The activity of glucose dehydrogenase present in resting spores of Bacillus subtilis varied strikingly with the conditions for disrupting the spores by sonic treatment, namely, the time and strength of sonication, and the type and pH of the solution used for suspending the spores. When the resting spores were sonicated for 30 min at a current of 1.45 A in 100 mm phosphate buffer in the range of pH 6.0 to 6.6 or in deionized water, the enzyme activity of the former suspension was approximately 10 times higher than that of the latter suspension. However, the enzyme activity of the latter was markedly stimulated in the presence of sodium chloride. The glucose dehydrogenase from resting spores disrupted in 100 mm phosphate buffer (pH 6.6) was a salt‐independent, active enzyme with a molecular weight of about 120,000, whereas the enzyme from resting spores disrupted in deionized water was a salt‐dependent, inactive one with a molecular weight of about 55,000. A high concentration of dipicolinic acid strongly inhibited activation by a salt of inactive glucose dehydrogenase from resting spores in deionized water, suggesting one of its several important roles in vivo.
Preparation of Membrane Fraction from Herpes Simplex Virus‐Infected Cells which Induce Cytotoxic T Lymphocytes
Authors: Hitsumoto Yasuo , Sonoda Shunro , Okuyama Masaaki , Miki Yoshiharu , Utsumi Sayaka
Published: November 2013

The immunogenic capacity of herpes simplex virs (HSV)‐infected cells and their subcellular membrane fractions was investigated by assessing the anti‐HSV cytotoxic T lymphocyte (CTL) response in cultures of spleen lymphocytes from HSV‐primed BALB/c mice. Methylchloranthrane‐induced fibrosarcoma (Meth A) cells infected with HSV (HSV‐Meth A) were fixed either with glutaraldehyde or by heating at 56 C to preserve their immunogenic competence and then used as a stimulator. Microsomes and plasma membranes were prepared from HSV‐Meth A and their immunogenic activities were determined. Though the recovery of stimulatory activity in the plasma membrane fraction was half of that in the microsome fraction, the activity in the former was much more stable than in the latter and the plasma membrane fraction proved to be well qualified as an immunogen for anti‐HSV CTL induction. Upon purification, the specific activity of the membrane fraction, on the basis of protein concentration, increased 43‐fold.
Suppression of Cell Mediated Immunity to Cytomegalovirus and Tuberculin in Pregnancy Employing the Leukocyte Migration Inhibition Test
Authors: Tanaka Akira , Hirota Kiyokata , Takahashi Katsuyuki , Numazaki Yoshio
Published: November 2013

To clarify the mechanism of reactivation of cytomegalovirus (CMV) in pregnancy, cell mediated immunity to CMV was investigated in 108 pregnant and 29 postpartal women employing the leukocyte migration inhibition technique. It was demonstrated that CMV‐specific cell mediated immunity was suppressed as gestation progressed in 20% of the seropositive women during the first trimester, in 78% during the second trimester and in all at term. The suppression of cell mediated immunity during pregnancy ceased 8 weeks after parturition. The results suggested that reactivation of CMV in pregnancy was probably caused by the suppression of CMV‐specific cell mediated immunity.
Drug Resistance and Conjugative R Plasmids in Fecal Escherichia coli Strains Isolated from Healthy Younger Animals (Chickens, Piglets, Calves) and Children
Authors: Kanai Hisashi , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

A total of 11,777 Escherichia coli strains were isolated from 90 chickens, 103 piglets, 96 calves, and 104 children in 1979 in Gunma Prefecture and tested for drug resistance and the presence of conjugative R plasmids. The percentages of individuals that excreted drug‐resistant strains were: chickens, 100%; piglets, 99%; calves, 100%; and children, 64%. The frequency of isolation of drug‐resistant strains among the total isolates was: chickens, 98%; piglets, 93%; calves, 94%; and children, 41%. Frequency of isolation of R plasmids among the strains tested was: chickens, 48%; piglets, 33%; calves, 38%; and children, 10%.

Resistance patterns of the strains isolated most frequently among the four groups were tetracycline (TC), sulfonamides (SA) in single resistance, TC.SA in double resistance, TC.streptomycin (SM).SA in triple resistance and TC.SM.SA. kanamycin (KM) in quadruple resistance.

R plasmids were isolated frequently from animals (over 33%) but infrequently from children (about 10%).

The high frequency of isolation of drug‐resistant strains and R plasmids from animals was caused by the heavy use of chemicals in the period of growth of younger animals.
Transformation of Human Embryonic Kidney Cells by a Viable Deletion Mutant of BK Virus
Authors: Hara Kazuya , Yogo Yoshiaki , Uchida Seijiro
Published: November 2013

Infection by a viable deletion mutant of BK virus (pm‐522) of human embryonic kidney (HEK) cultures originating from different fetuses generated cell lines, designated pmHEK, resistant to superinfection by the virus. However, infection of HEK cultures by a cloned wild‐type BK virus (wt‐501) did not generate a cell line. In medium with 10% calf serum all pmHEK cells reached saturation densities significantly higher than those of HEK cells and could grow in medium containing 1% serum. They did not form colonies in soft‐agar medium, and had limited life‐spans greatly extended beyond that of HEK cells. These results suggest that pmHEK cells are partially transformed. T antigen was uniformly expressed by all pmHEK cells, while V antigen was present in only a small minority of the cells. Thirty to 5,000 copies of the viral DNA per cell were detected, primarily in a nonintegrated form, in all pmHEK cell lines and the clones isolated from one of them. Since at least in two pmHEK lines, the actual quantities of free viral DNA per cell were sig‐nificantly greater than those estimated on the basis of the assumption that free viral DNA was produced only by the V‐antigen‐positive cells, it can be concluded that free viral DNA is also present in the V‐antigen‐negative cells predominant in the cell populations. Although the existence of a few copies of integrated viral sequences in pmHEK cells was not ruled out, the large amount of free viral DNA present in each cell probably plays a prominent role in the production of T antigen required for maintenance of transformation.
Immunogenic Dialyzable Factor Derived from a Ribosomal Fraction of Salmonella typhimurium
Authors: Kita Eiji , Matsuura Hiroshi , Masuda Shuji , Tomihata Shigeru , Kashiba Shuzo
Published: November 2013

The preparation, properties, and immunogenicity of the dialyzable factor from a ribosomal fraction of Salmonella typhimurium are described. The ribosomal fraction was purified to eliminate O‐antigenic components, by affinity chromatography (Sepharose‐anti‐O antibody conjugates used as immunoadsorbent). The dialyzable factor was obtained in the concentrated dialysate of the purified ribosomal fraction which was alternately frozen in dry‐ice acetone and thawed in an 80 C water bath, for a total of five or six cycles. When this preparation was tested for its ability to protect mice against challenge with 1,000 LD50 of the homologous bacteria, it afforded 100% protection at a dose equivalent to 5.0 μg of RNA. The protection conferred by this factor was mainly cell mediated but immune serum enhanced this immunity despite the fact that no antibodies were detected in it. The protective activity of this factor was sensitive to RNase digestion but resistant to proteolytic enzymes. Ion exchange chromatography of this factor with DEAE‐Sephadex A‐25 (in 7 M Urea‐0.02 M Tris‐HCl buffer, pH 7.5) resulted in a single A260 peak which was found to be immunogenic. Chemical analysis of this peak after it was concentrated and desalted revealed that this immunogenic fraction was composed mainly of mixed nucleotides. The data indicate that protective immunity conferred by a ribosomal vaccine is associated with RNA but may not require the intact RNA molecule.
Typing of Isolates of Mycoplasma pneumoniae and Mycoplasma salivarium by Growth Inhibitors Derived from Mycoplasmal Cells Treated with Chloroform
Authors: Nakamura Masahiro , Itoh Tohru
Published: November 2013

Isolates of Mycoplasma pneumoniae and M. salivarium could be subclassified at the strain level by inhibitors (ch‐Mcin) derived from mycoplasmal cells treated with chloroform. Sixty‐one isolates of M. pneumoniae obtained from oral cavities of patients were divided into three types by their ch‐Mcin: a definite type which completely inhibits the growth of M. fermentans PG18, an indefinite type and a no‐inhibition type. Sixty‐seven isolates of M. salivarium were also divided into similar types by their ch‐Mcin which does or does not inhibit the growth of M. salivarium Hup127. In the case of isolates of M. pneumoniae and M. salivarium belonging to the indefinite type which gave ambiguous patterns in the ch‐Mcin typing, it was demonstrated that they could be clearly typed by further testing after repeated cloning.
Effect of Yersinia enterocolitica ST on Cyclic Guanosine 3′, 5′‐Monophosphate Levels in Mouse Intestines and Cultured Cells
Authors: Inoue Takashi , Okamoto Keinosuke , Moriyama Takeshi , Takahashi Tetsuomi , Shimizu Kiyoji , Miyama Akio
Published: November 2013

We determined the effect of heat‐stable enterotoxin produced by Yersinia enterocolitica (Y. enterocolitica ST) on cyclic nucleotide levels in the intestines of 6‐day‐old mice and in cultured cell line cells. The concentration of cyclic guanosine 3′,5′‐monophosphate (cyclic GMP) in homogenates of the intestines increased four‐ to fivefold by 3 min after intragastric administration of 10 units of purified Y. enterocolitica ST. This increase continued for 60 min, and then the concentration of cyclic GMP fell toward the levels of the controls. On the other hand, fluid accumulation in the intestines was not evident until 60 min after administration of the toxin. Thus, the increase in intestinal cyclic GMP concentration preceded measurable fluid accumulation. The effect on both cyclic GMP levels and fluid accumulation was abolished by treatment of the ST with either alkali solution (pH 10.7) or 2‐mercaptoethanol. Likewise, cyclic GMP levels in cultured cells (CCL‐6, HeLa, L, and Mm‐1 cells) increased dose‐dependently by 10 min after incubation of the cells with the ST. Cyclic adenosine 3′,5′‐monophosphate levels in both intestines and cultured cells were not affected by the toxin.
Analyses of Idiotypes on Anti‐α‐1, 6‐Dextran Antibodies in Mice
Authors: Haba Seiji , Hayashi Masue , Matsuoka Yuji
Published: November 2013

When mice of strains C57BL/6, C3H/He, and BALB/c were immunized with native dextran B512, only a small amount of IgM antibody was produced, but a substantial amount of anti‐dextran antibody of IgG class was produced after immunization with a conjugate of dextran T10 and keyhole limpet hemocyanin regardless of the mouse strain used. Isoelectric focusing (IEF) spectra revealed limited heterogeneity of anti‐dextran antibody of IgG class with strict consistency in all individual sera from C57BL/6 mice, even after secondary immunization, whereas antibodies from C3H/He and BALB/c mice showed more heterogeneous IEF spectra with some individual variations.

Rabbit anti‐idiotypic (Id) antibodies were raised by immunization with a subfraction of anti‐dextran antibody of IgG class from C57BL/6 mice, which showed major bands focused at around pH 7.7 upon IEF. It was found by using the anti‐Id antibodies that virtually all anti‐dextran antibody molecules of both IgG and IgM classes from C57BL/6 mice possessed common Id determinants which can be classified into two specificities, one specific for antibody from C57BL/6 mice and the other cross‐reactive with antibodies from BALB/c and C3H/He mice. About 80% of the antibody molecules from BALB/c and less than 20% of those from C3H/He mice were positive for the interstrain cross‐reactive Id. Both Id determinants seemed to be closely related to the antigen binding sites, or at least to reside in the vicinity of the antigen binding sites of anti‐dextran antibody.
Variation in Virulence of Coxsackie Virus B3 in the Hearts of Mice
Authors: Komatsu Toshihiko , Hashimoto Isao , Kohara Teruko
Published: November 2013

Virulence of Coxsackie virus B3 (CB3) was compared in mice between strain SK‐74 isolated from a patient and strain T‐70 isolated from a healthy child as well as between prototype strain Nancy and its mouse‐passaged derivative strain PMH. Strain SK‐74 showed a high mortality (40–80%), while strain T‐70 did not induce deaths in mice (mortality: 0%). Strain PMH showed a high mortality (65–85%), but strain Nancy did not cause deaths in the mice. In agreement with the mortality trend, virus titers in the heart and other organs of mice inoculated with strain SK‐74 and PMH were higher than those in mice inoculated with T‐70 and Nancy strains. Since virus titers in the heart remained higher than those in other organs, the heart was regarded as the main organ in which CB3 could replicate and induce cell degeneration. The present results suggest that a marked variation in virulence in the hearts of mice among CB3 strains occurred and that passage through the hearts of mice enhances the virulence of CB3 in the hearts of mice.