bluuebunny/crossref_metadata_2025 huggingface dataset:
Anaerobic Coryneforms Isolated from Human Bone Marrow and Skin
Authors: Saino Yushi , Eda Junji , Nagoya Takao , Yoshimura Yoko , Yamaguchi Masahito , Kobayashi Fujio
Published: November 2013

Eighteen isolates of anaerobic coryneforms from human bone marrow and skin and four type strains of Propionibacterium were studied chemically, biochemically and antigenically. All of the isolates were identified as Propionibacterium acnes; of the 18 isolates, 16 belonged to serotype I and two to serotype II. By means of gas liquid chromatography and mass spectral analysis, a large amount of iso‐type fatty acids, such as iso‐pentadecanoic and iso‐heptadecanoic acids were detected in whole cells of isolates and type strains. Antitumor and adjuvant effects of the isolates and type strains were found to differ considerably among the strains. One of the isolates, P. acnes C‐7, which showed potent biological activities was fractionated by hot phenol‐water extraction. The resulting insoluble middle layer was found the most effective in tumor protection, adjuvant action in immune response and phagocytic activity in mice.
Immunosuppressive Effect of Bacterial Lipopolysaccharide on Antibody Response
Authors: Nakano Masayasu , Tanabe Masao J. , Saito Tatsuo , Shimizu Tadayori
Published: November 2013

Injection of endotoxins (bacterial lipopolysaccharide: LPS) several days prior to immunization causes the suppression of antibody response. The suppressive effects of several kinds of LPS preparations on the plaque‐forming cell (PFC) antibody response in the spleen of mice were examined after immunization with sheep red blood cells (SRBC). Glycolipids obtained from heptoseless mutants (Re form) of salmonella or its lipid A preparation coupled artificially with bovine serum albumin (BSA) are capable, like LPS obtained from a wild type (S form) strain, of inducing suppression of the PFC response, while alkaline‐detoxified LPS can not. The refractory periods of the PFC response induced by LPS injection last only a few days. However, the use of cyclophosphamide (CY) together with LPS can extend the refractory periods of antigenic stimulation for several weeks. Injections of LPS and CY can also induce unresponsive states of OH agglutinin antibody response to antigenic stimulation with formalin‐killed organisms of Escherichia coli or Salmonella enteritidis (presumably both thymus‐independent antigens). These unresponsive states induced by LPS and CY are easily terminated by a transfer of syngeneic bone marrow cells but not by thymocyte transfer.
Type and Strain Variation in Induction of L Forms of Neisseria gonorrhoeae
Authors: Ota Fusao , Ashton F.E. , Diena B.B.
Published: November 2013

Strains of Neisseria gonorrhoeae were inoculated onto brain heart infusion (Difco) agar supplemented with penicillin and polyvinylpyrrolidone (PVP) as a stabilizer and cultivated in a candle extinction jar. L‐form colonies became visible after a few days. They continued to grow and were viable for up to 38 days. The extent of inducibility of L forms of N. gonorrhoeae was examined semiquantitatively. It was found to be constant for each type and strain and to depend only slightly on the amount of penicillin added to the medium. None of the types of one strain produced L‐form colonies. In another strain, avirulent types(T3, T4) showed more ability to produce L forms than virulent types (T1, T2) and no L forms were produced by the subtypes of T1 and T2–T1a and T2a. In a third strain, only T4 produced L forms. Electron microscopic studies showed that the L forms consisted of a number of membranous vesicles and a variety of cell types such as those completely lacking cell walls and those with only remnants of cell walls. The results indicate the existence of subtypes of T1 and T2 of gonococci and the intrinsic inducibility of gonococcal types and strains to produce L forms.
Interaction of Human Waldenström's IgM Proteins with Guinea Pig and Human Complement1
Authors: Ikeda Masaharu , Onoue Kaoru
Published: November 2013

The activity of purified human Waldenström's IgM proteins to fix complement of human and guinea pig origins was compared at different temperatures using the polystyrene latex particle‐adsorption method. It was shown that the interaction of the IgM proteins with complement differed depending on the source of complement and that a pronounced heterogeneity in complement‐fixing activity was observed among the IgM proteins when tested with guinea pig complement. Thus, by the use of guinea pig complement, six human IgM proteins examined were classified roughly into two groups, one having a high and the other a low activity at 3 C as well as at 37 C. With human complement, five proteins showed a rather uniform activity at 37 C. However, there was one protein with no detectable activity, suggesting the presence of non‐complement‐fixing protein in the IgM class. All the six proteins showed no significant activity with human complement at 3 C. No antigenic difference has been found as yet in the Fc or Cμ2 region among these IgM proteins examined.
Microorganism Capable of Decomposing N‐Acetylglucosaminyl Ribitol Teichoic Acid of Staphylococcus aureus
Authors: Iwata Sadako , Tochikubo Kunio , Kato Keijiro , Hirata Takanori , Kotani Shozo , Yagi Kunio
Published: November 2013

Enzyme(s) capable of decomposing N‐acetylglucosaminyl ribitol teichoic acid prepared from the cell wall of Staphylococcus aureus FDA 209 P was obtained from the culture supernatant of a gram‐negative, rod‐shaped, spore‐forming soil bacterium. Properties of the bacterium were very similar to those of Bacillus circulans.
Behaviors of a Vigorous Protease‐Producing Strain of Staphylococcus aureus in the Skin Tissue of Mice
Authors: Takeuchi Syotaro , Suto Tsuneji
Published: November 2013

The behaviors of a protease‐producing strain of Staphylococcus aureus, including the production of α‐ and β‐hemolysin, protease, and nuclease in the skin tissue of mice, were examined by the fluorescent antibody technique and hematoxylin and eosin staining. When about 106 viable cells were subcutaneously inoculated into a mouse, they were phagocytized by leukocytes and a suppurative focus developed in situ. No active extracellular substances other than α‐hemolysin were demonstrated in this mouse. When 107 viable cells were inoculated, most of them were also phagocytized by leukocytes, and a rather large suppurative focus was formed. In this focus a low degree of multiplication of the organisms was observed. Protease and α‐hemolysin could be detected in the neighborhood of the leukocytes. When 108 to 109 cells were inoculated, they multiplied transitorily in the subcutaneous tissue and produced the four substances mentioned above. The multiplication of bacterial cells as well as the production of extracellular substances ceased within a comparatively short period and the transitory state was similar to that in a batch fermenter. Necrotic and histolytic lesions were observed only in the mice inoculated with 108 to 109 viable cells. Similar dermatolytic lesions were also found in mice injected with 0.5 to 1.0 mg of pure protease.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Kato Nobuo , Nakashima Izumi , Ohta Michio
Published: November 2013

The neutral fraction (neutral CPS‐K) of Klebsiella pneumoniae capsular polysaccharide (CPS‐K) from type 1, Kasuya strain, has already been reported as the active substance responsible for the strong adjuvant effect of CPS‐K. The present results demonstrate that neutral CPS‐K exhibits further common biological activities with lipopolysaccharide (LPS) isolated from Salmonella enteritidis. The intensity of the lethality in mice of neutral CPS‐K by the intraperitoneal route is very similar to that of LPS. Its lethality for mice by the intravenous (i.v.) route is significantly stronger than that of LPS, because the degree of increase in the sensitivity to their lethality by i.v. challenge is smaller for LPS than for neutral CPS‐K. The intensity of the pyrogenicity of neutral CPS‐K in rabbits is approximately one‐tenth of that of LPS as judged by the minimal pyrogenic doses and fever indices. The skin‐preparatory potency of neutral CPS‐K for the dermal Shwartzman phenomenon in rabbits is also approximately one‐tenth of that of LPS compared on the basis of the minimal skin‐preparatory doses. When injected i.v., neutral CPS‐K exhibits a provocative effect on hemorrhagic reactions in skin sites prepared with neutral CPS‐K or LPS.
Studies on Delayed Hypersensitivity of Antigens Isolated from Mycoplasma pneumoniae Cells
Authors: Suzuki Masuko , Hayashi Yoshiro , Arai Sumio , Kumagai Katsuo
Published: November 2013

Cells of Mycoplasma pneumoniae Mac strain were fractionated into acetone‐soluble and insoluble fractions. Acetone‐insoluble fractions were digested with pronase and further purified by chromatography on Sephadex G‐75, yielding three water‐soluble fractions which were free from lipid and consisted mainly of polysaccharide‐protein complex. All these water‐soluble fractions possessed eliciting antigenicity to delayed hypersensitivity for M. pneumoniae as measured by skin reactions and macrophage migration inhibition tests, but not to complement‐fixing antibodies. In contrast, the acetone‐soluble fraction was reactive with the complement‐fixing antibodies but not for the delayed hypersensitivity.
Preferential Phosphorylation of NP‐Protein of Influenza A2 Virus by Virion‐Associated Protein Kinase
Authors: Sugiyama Kazuo , Kamada Tohru , Shimizu Kazufumi , Watanabe Yasushi
Published: November 2013

Influenza A2 virions were found to contain protein kinase activity which was stimulated, like in other virion‐associated kinases, with Mg++ and Nonidet‐P 40 but not with cyclic AMP. The kinase phosphorylated only the NP‐protein fraction of the influenza virions in the in vitro reaction. In contrast, none of the influenza virion proteins were phosphorylated significantly during the process of virus production in infected chorioallantoic membranes, The in vitro and in vivo phosphorylations of influenza viral proteins were compared with those of Sendai virus (HVJ).
Transition in the Character of Immunological Memory in Mice after Immunization
Authors: Takaoki Muneo , Kawaguchi Susumu , Katsura Yoshimoto , Muramatsu Shigeru
Published: November 2013

The immunological memory in antibody response of mice to bovine serum albumin was investigated at the level of IgM and IgG antibody‐forming cells. The antigen at a dose much lower than required for eliciting a detectable level of the primary antibody response could latently activate the immune machinery to an extent adequate for specific recall, whereas higher doses of antigen were effective in evoking strong anamnestic response. The potentiality to develop the anamnestic response was found even in the latent phase of the primary antibody response and was maintained for more than 2 months. The immunological memory acquired in an early phase after the primary immunization mainly involved IgM antibody response and late memory concerned IgG response.
Immune Response against Hamster Erythrocytes in the Low‐Responder Mouse Strains
Authors: Nomoto Kikuo , Harada Tatsushi , Koyanagi Rieko , Hosoi Masaharu , Takeya Kenji
Published: November 2013

Enhancing and suppressing effects of microbial adjuvants were studied in female mice of the C3H/He, AKR and SL strains. Propionibacterium acnes, Bordetella pertussis, BCG and yeast cell wall (YCW) were chosen as adjuvants. As antigens, we chose hamster erythrocytes (HRBC) which proved to be a weak antigen for mice. Adjuvants were given on day —7, day 0 or day 3, and HRBC were injected on day 0. The results were as follows. 1) P. acnes facilitated IgM and IgG antibody production in AKR mice and suppressed IgM antibody production in SL mice, when given on day —7. When P. acnes was given on day 0, they suppressed IgM antibody production in all of the strains used. 2) When B. pertussis was given on day 0, it exhibited enhancing effects on IgG antibody production in all of the strains and a suppressing effect on IgM antibody production in SL mice. 3) BCG suppressed IgM antibody production in all strains when given on day 0. 4) YCW showed no influence on antibody production in any combination used in this work. 5) SL mice were very sensitive to suppressing effects by adjuvants. Strain differences in the expression of enhancing and suppressing effects by adjuvants appear to be under some control independent of antigen‐specific immune response genes.
Inhibition of Intracellular Multiplication of Adenovirus by Interaction between Infected and Uninfected Cells
Authors: Imai Masao , Hamada Chuya , Uetake Hisao
Published: November 2013

The possibility that virus multiplication may be inhibited by interaction of infected cells with uninfected cells was tested by experiments, using human adenovirus type 12 (Ad 12). Permissive human cells (human embryonic kidney = HEK, KB or HeLa) were infected and seeded on uninfected or infected “nonpermissive” cell (human embryonic lung = HEL) monolayers, and virus yields or proportions of viral antigen‐synthesizing cells were compared with each other. Both the virus yields and the proportions of viral antigen‐positive cells were not reduced significantly when seeded on infected HEL cells, while when seeded on uninfected HEL cells both of them were reduced remarkably, compared with the yield and the proportion of controls seeded on glass. Similar results were obtained regardless of the type of permissive cells, HEK, KB, or HeLa. Similar reduction of the yield was observed when seeded on HEL cells infected with Ad 12 inactivated by heat or by antiserum, and partial reduction was observed when seeded on HEL cells infected with UV‐inactivated Ad 12, depending on the extent of UV dosis. These experiments showed that intracellular virus multiplication may be inhibited by interaction of infected cells with uninfected cells, and this may be due to the difference in the cell surface structure.
Mechanisms of the Adjuvant Effect of Nystatin on In Vitro Antibody Response of Mouse Spleen Cells
Authors: Ishikawa Hiromich , Narimatsu Hisashi , Saito Kazuhisa
Published: November 2013

Adjuvanticity of nystatin, one of the polyenic antifungal antibiotics having as its primary target the membrane sterol of eukaryotic cells, was investigated by examining its effect on several functions of mouse spleen cells relevant to immunological phenomena in vitro. Nystatin was found to stimulate significantly DNA synthesis in thymus‐independent (B) cells but not in thymus‐dependent (T) cells. Like the other B‐cell mitogens such as bacterial lipopolysaccharide (LPS), nystatin elicited nonspecifically polyclonal antibody synthesis in mouse spleen cell cultures, and also restored antibody response of T cell‐deficient spleen cells of congenitally athymic nude mice to heterologous erythrocytes (RBC; thymus‐dependent antigen). Thus, nystatin and LPS appeared to cause similar changes in the functions of spleen cells relevant to immunological events. However, antagonism but no additive effect in the adjuvanticity was revealed between the two adjuvants. As an interesting finding, the polyclonal generation of anti‐RBC antibody‐forming cells (AFC) in the spleen cell cultures by stimulation with B‐cell mitogen, i.e., either nystatin or LPS, was not inhibited at all by inclusion of any anti‐RBC antiserum, whereas, as is well known, the generation of AFC by stimulation with the antigen was specifically suppressed by the corresponding antiserum, indicating a difference in the genesis between the mitogen‐induced AFC and the antigen‐induced AFC.
Difference in Genesis between Antigen‐Induced IgM Antibody‐Forming Cells and B‐Cell Mitogen‐Induced IgM Antibody‐Forming Cells in Mouse Spleen Cell Cultures
Authors: Ishikawa Hiromichi , Imanishi Kenichi , Saito Kazuhisa
Published: November 2013

To determine the mechanisms in the triggering of thymus‐independent lymphocytes (B cells) for development into antibody‐forming cells (AFC), genesis of IgM AFC elicited polyclonally by nonspecific stimulation with B‐cell mitogen, such as nystatin and bacterial lipopolysaccharide, was compared with that of IgM AFC specifically elicited by antigenic stimulation, using mouse spleen cell cultures as an experimental system and sheep erythrocytes (SRBC) as a test antigen. Considering that differentiation and proliferation are necessary cellular events for precursor B cells to develop into AFC, the effect of different antimetabolic agents on the generation of each type of AFC in spleen cell cultures was examined. The generation of anti‐SRBC IgM hemolysin plaque‐forming cells (PFC) in B‐cell mitogen‐stimulated spleen cell cultures was found to be less susceptible to X‐irradiation or mitomycin C than that in the SRBC‐stimulated cultures. These apparently paradoxical results were affiirmed using colcemid as an inhibitor of cell mitosis and hydroxyurea (HU) as an inhibitor of cellular DNA synthesis. Thus, when spleen cell cultures responding to either SRBC or B‐cell mitogen were exposed to colcemid or HU during a period from 2 days to 3 days after the stimulation, the exponential generation of anti‐SRBC IgM PFC in the cultures responding to SRBC was completely halted, whereas that in the cultures responding to B‐cell mitogen was not. Furthermore, N6, O2′ ‐dibutyryl adenosine 3′, 5′ ‐cyclic monophosphoric acid was found to halt the exponential generation of antigen‐induced anti‐SRBC IgM PFC but not that of the B‐cell mitogen‐induced anti‐SRBC IgM PFC. From these results it was suggested that B‐cell mitogen might stimulate precursor Bμ cells at a late stage in the differentiative pathway to develop into AFC without cell division, and that antigenic stimulation might stimulate relatively primitive precursor Bμ cells to proliferate and then differentiate into AFC. Based on this idea, mechanisms in the triggering of B‐cell activation are discussed.
Toxin Isolation from a Kanagawa‐Phenomenon Negative Strain of Vibrio parahaemolyticus
Authors: Sochard M.R. , Colwell R.R.
Published: November 2013

Production of a toxin by Vibrio parahaemolyticus Kanagawa‐phenomenon negative strains was examined. Ammonium sulfate fractions of broth culture filtrates were dialyzed, concentrated by lyophilization, and tested for toxic effects by mouse intraperitoneal injection. One fraction, which we think is a toxin, was isolated from a broth culture filtrate of V. parahaemolyticus FC 1011 (a Kanagawa‐phenomenon negative strain) and consistently produced lethal effects in mice at high concentrations and diarrhea in lower concentrations. The toxin was assayed for mouse LD50 and ability to produce diarrhea via forced feeding in mice. V. parahaemolyticus FC 1011 toxin was found to be protein, to be inactivated by heat or trypsin hydrolysis, and to produce positive skin permeability reactions in rabbits. However, it failed to induce fluid accumulation in ligated ileal loops in rabbits.
Antigenic Variation of Poliovirus Caused by Antibody Components with Different Specificities
Authors: Urasawa Shozo , Ishizawa Fumihiko , Urasawa Tomoko
Published: November 2013

The possible role of antibody as a selective pressure on antigenic mutants of poliovirus in nature was investigated in vitro. A mutant resistant to a monospecific antibody with a definite specificity was readily obtained by several cycles of neutralization of Mahoney strain with a monospecific antibody and multiplication in monkey kidney (MS) cells. Mutants resistant to more than two different monospecific antibodies were also readily obtained in a similar manner. Studies on the antigenicity of these mutants by kinetic neutralization tests revealed that the Mahoney strain underwent a progressive serological variation as it became successively resistant to one to five different monospecific antibodies isolated from anti‐Mahoney serum.
Induction and Properties of DNP‐Specifie T Cells in Mice Sensitized to DNP‐Coupled Mycobacterium
Authors: Hirai Yoshikatsu , Kishimoto Tadamitsu , Suemura Masaki , Yamamura Yuichi
Published: November 2013

Splenic T lymphocytes from mice sensitized to 100 μg of DNP‐coupled mycobacterium (DNP‐Tbc) showed in vitro proliferative response against DNP‐or TNP‐conjugated protein antigens. The increased uptake of 3H‐thymidine induced by DNP‐HSA was partially inhibited by the addition of 10–4M DNP‐EACA. DNP‐AECM‐Ficoll did not induce any significant proliferative responses in DNP‐Tbc‐primed T cell population. However, priming with DNP‐Tbc augmented anti‐DNP IgG antibody response induced with DNP‐Ficoll. The augmentation of IgG response was not due to the presence of DNP‐primed B cells or anti‐DNP antibody. The results showed that the priming with DNP‐Tbc induced DNP‐reactive T helper cells which could be triggered with DNP‐Ficoll. The possible role of mycobacterium in the induction of hapten‐specific T helper cells is discussed.
Studies on the Pathogenicity of Yersinia enterocolitica
Authors: Une Tsutomu
Published: November 2013

Comparative studies on pathogenicity between Yersinia enterocolitica and Yersinia pseudotuberculosis were performed using experimental infection systems in vivo and in vitro.

All of thestra ins of both species successfully produced experimental enterocolitis in rabbits although the severity varied with the strains challenged. The changes were characterized by granulomatous lesions with necrobiotic centers in reticuloendothelial tissues of the intestine, mesenteric lymph nodes, liver and spleen.

These strains uniformly had the ability to penetrate HeLa cells and to survive or multiply within cultured rabbit peritoneal macrophages.

In addition, in infections with strain TP‐2 or PST‐III of Y. pseudotuberculosis, catarrhal inflammation all over the small intestine and/or focal necrosis and parenchymatous degeneration in the liver were observed, along with the granulomatous lesions. These strains, at the same time, exhibited cytotoxic effects on the cultured cells.

The pathogenic factors of Y. enterocolitica are discussed in comparison with those of Y. pseudotuberculosis.
Effects of Lectins on the Hemolysis of Rabbit Erythrocytes by Staphylococcal Alpha Toxin
Authors: Kato Iwao , Sakoda Keiko , Saito Masaru , Suzuki Yoshie
Published: November 2013

When concanavalin A (1 μg/ml) or wheat germ agglutinin (2 μg/ml) was preincubated with a suspension of 2% rabbit erythrocytes for 5 min at 20 C, the binding of [125I]‐labeled staphylococcal alpha toxin to these erythrocytes was greatly inhibited and the hemolytic action of alpha toxin was decreased. The inhibitory effect of concanavalin A on hemolysis by alpha toxin was completely reversed in the presence of 0.1 M α‐methyl‐D‐glucoside or α‐methyl‐D‐mannoside. Phytohemagglutinin‐P from Phaseolus vulgaris and soybean agglutinin inhibited hemolysis by the toxin at concentrations exceeding 20 μg/ml.

The effect of concanavalin A on alpha‐toxin hemolysis was studied further to ascertain the nature of the inhibition. Double reciprocal plots were made of hemolysis against alpha toxin concentrations, and the data suggested that inhibition of the initial rate of the hemolysis by concanavalin A is competitive in nature. This was probably due to an interaction with the alpha toxin binding sites on the cell membrane surface.
Electron Microscopic Study of Hemadsorption on Vaccinia Virus Infected Cells
Authors: Saburi Yasuo , Okuda Kenji , Takahashi Toyozoh , Tadokoro Ichiro
Published: November 2013

Hemadsorption (HAD) induced in HEp‐2 cells infected with vaccinia virus was observed. In ultrathin sections, binding of 36 red blood cells (RBCs) was examined in detail and 3 types of HAD were observed: (1) direct and close binding of RBCs to infected HEp‐2 cells (cyto‐HAD) was observed in cross sections of 27 RBCs, (2) binding of RBCs through microvilli of infected cells was found in 11 RBCs, and (3) five RBCs were distorted to form tentacle‐like projections by which they were bound to the HEp‐2 cell surface. Scanning electron microscopy revealed that more than 30% of the RBCs were bound to microvilli of vaccinia virus‐infected HEp‐2 cells, and that the number of microvilli twined round each RBC was over ten. RBCs were attached to certain microvilli through swollen sucker‐like tips which were not observable in non‐infected HEp‐2 cells. RBCs sometimes revealed a polygonal shape at regions of binding to microvilli. Virion‐mediated RBC‐HEp‐2 cell binding could not be observed.
Immunological Properties of Vibrio cholerae Lipopolysaccharides
Authors: Nakano Masayasu , Tanabe Masao J. , Hori Hideko , Kondo Seiichi , Hisatsune Kazuhito
Published: November 2013

Immunological effects of wall lipopolysaccharide (LPS) preparations obtained from Vibrio cholerae Inaba 569B, Ogawa NIH 41 and NAG 4715 strains by the hot phenol‐water procedure were examined in mice. Although these LPS lack KDO, which are basic components of the core region of most gram‐negative LPS, they still have potencies as B‐cell mitogens, adjuvants, immunosuppressants, polyclonal B‐cell activators and phagocytic stimulants for macrophages. The activities of these V. cholerae LPS on murine immune system seemed to be weaker than those of Salmonella typhimurium LT2‐LPS. Among these V. cholerae LPS, NAG 4715‐LPS showed the strongest mitogenic activity and phagocytic stimulation, while the potencies of this NAG 4715‐LPS for the induction of polyclonal B cell activation, adjuvant effects and immunosuppression did not seem to be greater to those of the other LPS.
Temperature Sensitive R Plasmids Isolated from Proteus Strains
Authors: Odakura Yoshihiro , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Out of 32 R plasmids isolated from Proteus strains, 17 were found to be temperature sensitive with respect to inheritance in E. coli cells. They were fi– and classified into incompatibility group T or V. Cells carrying T group Rms273 plasmid were temperature sensitive with respect to growth and conjugal transfer in both E. coli and Proteus. The V group YOR‐10 plasmid was stable in Proteus even at 42 C. However, the loss frequency of YOR‐10 plasmid in E. coli reached 100% after 4 hr of incubation at 42 C, in spite of stable inheritance at 25 C. Conjugal transfer of the YOR‐10 plasmid in E. coli was also strongly inhibited at 42 C. It has been concluded that instability of V group R plasmids in E. coli is due to their thermosensitive inheritance in the progeny cells at high temperatures.
Studies on the Minimum Reproducible Unit of Staphylococcal L‐Forms
Authors: Okuda Kenji , Takahashi Toyozoh , Tadokoro Ichiro , Eda Tohru
Published: November 2013

The minimum size of a reproducible unit of staphylococcal L‐forms was determined by filtration and electron microscopic methods. Ultrathin sections of an induced strain of Staphylococcal L‐forms (STA‐EMT‐1) in liquid medium revealed several types of structures, all of which were bound by a single membrane and most of which possessed ribosome‐like granules. Many of the small granules were less than 0.3 μm and were attached to the membrane of the large bodies. Using a serial filtration method, it was observed that viable L‐forms were still detected in 0.22 μm filtrate, but the viable cell count of L‐forms decreased in number with the decrease in pore size of membrane filters. A fractionation technique, using L‐forms filtered through a membrane filter with a 0.45 μm pore size, revealed that there were three classes of small bodies but only the first class with ribosome‐like granules over approximately 0.2 μm in diameter seems to be able to reproduce.
Immunological Properties of Propionibacterium Acnes
Authors: Nagoya Takao , Kobayashi Fujio , Nomoto Kikuo
Published: November 2013

Adjuvant activity of phenol‐treated cells of Propionibacterium acnes C‐7 in antibody response was investigated in ICR mice. Simultaneous administration (day 0) of P. acnes (i.p.) and sheep red blood cells (SRBC) (i.v.) enhanced the formation of direct plaque‐forming cells (PFC) on day 2, and the formation of indirect PFC response on day 7 and thereafter. Conversely, pretreatment from 11 to 14 days before antigen injection suppressed markedly the antibody response. The potentiation and the suppression of immune response depended on doses of antigen and of P. acnes, the timing of adjuvant injection and the time of assay. The two opposite phenomena caused by P. acnes were also confirmed in antibody response against hamster red blood cells (HRBC). Pretreatment with P. acnes 1 to 14 days before antigen injection suppressed markedly anti‐HRBC antibody response, whereas P. acnes injected simultaneously with HRBC or one day after injection of the antigen induced prolongation of antibody response and the production of 2‐mercaptoethanol‐resistant antibody.
In Vivo Formation of Transmissible Resistance Factor by Recombination between Nontrans‐missible Resistance Factor and Col B Factor
Authors: Kameda Mitsuo , Suzuki Kaname , Mitsuhashi Susumu
Published: November 2013

Germ‐free swine were artificially contaminated with tetracycline (TC) sensitive strains of Escherichia coli and Klebsiella pneumoniae. One of these strains, E. coli 3306, was infected with a plasmid carrying kanamycin (KM) resistance, i.e., T‐kan factor. Another strain, E. coli P‐5, carried a conjugally transferable Col B factor. Among the nine strains used, only E. coli P‐38 became TC‐resistant after TC administration. Three types of TC‐resistant E. coli P‐38 strains were found; (a) one strain carried nontransferable TC resistance and could not produce colicin, (b) one strain carried TC resistance with a high transmission frequency which could not produce colicin, and (c) one strain carried TC resistance with a low transmission frequency that could produce colicin B. Genetic studies disclosed that the transmissible TC resistance factors, i.e., Rms105 (group b) and Rms104 (group c), were formed by recombination between Col B factor and nontransmissible TC‐resistance (let) determinant which appeared in E. coli P‐38 mutants.
Acquired Immunity to Mycoplasma pneumoniae Pneumonia in Hamsters
Authors: Hayatsu Eizo
Published: November 2013

An inactivated Mycoplasma pneumoniae vaccine was prepared from a culture in a liquid medium supplemented with water extract of egg yolk. Vaccinated Syrian hamsters were exposed to virulent M. pneumoniae aerosol and were examined for the retention of mycoplasmas and for histopathological changes in the respiratory tracts. When a vaccine prepared with strain FH was administered intramuscularly or by inhalation in aerosol, no significant resistance was shown with respect to mycoplasma proliferation. An increased resistance, however, was observed when an aluminium phosphate‐adsorbed vaccine, and when a plain vaccine (although to a lesser degree) prepared with hamster 24‐passaged strain FH, was administered intramuscularly. Histopathologically, lung lesions were markedly suppressed in groups showing high resistance. A correlation between the serum antibody titer and the resistance to infection was observed. Hamsters which received a hyperimmune rabbit antiserum intracordally showed a high resistance to M. pneumoniae infection. The suppression of histopathological changes also coincided with high complement‐fixing antibody titers of either actively or passively immunized hamster serum. The results suggest that humoral immunity plays an important role in resistance to M. pneumoniae pneumonia in hamsters.
Behavior of a Vigorous α‐ or β‐Hemolysin‐Producing Strain of Staphylococcus aureus in the Cutaneous Tissue of Mice
Authors: Takeuchi Shotaro , Nakajima Yasuyuki , Shoya Shigemi , Suto Tsuneji
Published: November 2013

Two strains of Staphylococcus aureus were examined for behavior in the cutaneous tissue of mice by the fluorescent antibody technique, hematoxylin and eosin staining. When about 108 viable cells of an α‐hemolysin‐producing strain (Wood 46) were inoculated subcutaneously into a mouse, they multiplied in the subcutaneous tissue of the mouse and gradually entered the corium to produce α‐hemolysin and nuclease. Edematous and necrotic lesions were observed in the cutaneous tissue where the organisms had multiplied. When 108 viable cells of a β‐hemolysin‐producing strain (Kitami 3–9D) were inoculated into a mouse, they multiplied within a narrow extent surrounded mainly by infiltrating leukocytes and produced mainly β‐hemolysin. The changes of cutaneous tissue were weaker in mice inoculated with Kitami 3‐9D strain than in mice inoculated with strain Wood 46. When 106 viable cells of both strains were inoculated into mice, they were phagocytized by leukocytes. Neither multiplication of organisms nor production of any active extracellular substance was observed in these mice. Edema, degeneration, and necrosis were also noticed in the cutaneous tissue of mice inoculated with α‐ and β‐hemolysin. In addition, the infiltration of leukocytes was inhibited mainly by α‐hemolysin.
Immune Reconstitution in the Immunodeficient Chicken by Transfer of Embryonic Lymphoid Cells
Authors: Moriya Osamu , Ichikawa Yoichi
Published: November 2013

Embryonic chickens were rendered immunodeficient by in ovo injection of homologous IgM on the 10th embryonic day. The immunodeficient embryos were intravenously given lymphoid cells taken from normal embryonic bursa, spleen or thymus on the 15th embryonic day. Gain of splenic or bursal weight, natural antibodies to sheep red blood cells (SRBC), frequencies of rosette forming cells (RFC) binding to SRBC or dinitrophenyl‐SRBC (DNP‐SRBC) and of immunoglobulin bearing cells (IBC) in the bursa and the spleen were investigated to assess the effect of transferred cells during the embryonic stage. Transferred bursal and splenic cells showed an ability to restore the frequency of RFC or IBC in the recipients. However, reversion from the immunodeficient state was not observed in the thymic cell transfer. These findings suggested that the cells derived from embryonic bursa and spleen contained stem cells which developed into RFC and also into precursors of IBC.
Age Related Differences in Cellular Immune Response to Vaccine Induced Rubella Infection
Authors: Chiba Yasuo , Fitzpatrick Paul , Patel Pratibha , Scott Ronald , Ogra Pearay L.
Published: November 2013

Employing the techniques of in‐vitro lymphocyte transformation (LTF) and using Putnam strain of rubella virus as the antigen, the development of rubella specific cellular immune response was studied in different age groups of rubella seronegative normal subjects at various intervals after subcutaneous administration of HPV‐77/DE5 live attenuated rubella vaccine. The rubella specific lymphocyte response in children ranging in age from two to twelve years was characterized by significant LTF activity at two months, followed by a gradual decrease. The response in adult subjects 18 to 35 years of age showed a slight delay initially in the appearance and the maximum LTF activity appeared to be 3–4 fold lower (P < 0.01) than observed in the children. No difference was observed in the maximum antibody titers to rubella virus between these two groups of subjects. These observations suggest that the age related differences in the lymphoproliferative responses might be associated with adverse effects which are known to occur more frequently in adolescent and older patients than in childhood population after vaccine induced rubella infection.
Genetic Control of Granuloma Response to Oil‐Associated BCG Cell Wall Vaccine in Mice
Authors: Yamamoto Ken‐Ichi , Kakinuma Mitsuaki
Published: November 2013

Intravenous injection of mice with BCG cell wall vaccine induces granuloma in the lung. The granuloma induced as measured by lung weight showed marked strain differences; C3H/HeMs bred in this Institute (C3H), C3HeB/FeJ, P/J and A/He were low responders, whereas C57BL/6 and AKR/He were high responded. When C57BL/6 and C3H were compared, the responsiveness appeared to be inherent in each strain of mice providing strong evidence for genetic control in this phenomenon. Breeding experiments suggested that multiple loci, including a MHC‐linked locus, were involved although the role of the MHC‐linked loci was marginal.
Cell‐Mediated Immunity to Mouse Adenovirus Infection
Authors: Inada Toshiki , Uetake Hisao
Published: November 2013

Cell‐mediated immunity (CMI) to mouse adenovirus (M‐Ad) infection was studied by macrophage migration inhibition test (MMI) as one of in vitro correlates of CMI.

Both direct and indirect tests showed clearly that migration of packed peritoneal exudate cells (PEC) (immune mouse or nonimmune guinea pig) was remarkably inhibited; MIF was produced by interactions between immune PEC and infected cell extracts and between immune spleen cells and infected cells or their extracts. The antigen(s) responsible for the above MMI was demonstrated in 6‐ to 12‐hour infected ME cells, and FUdR‐treated infected ME cells. Since under these conditions there is S antigen(s) synthesis but not capsid antigen synthesis, the antigen(s) concerned must be an S antigen(s). T cells sensitized to infected cells were shown to be required to induce MMI.

The MMI is specific for M‐Ad, since no cross MMI was observed between M‐Ad and SV40 systems. Time course study of the development of CMI to M‐Ad by MMI tests showed that CMI became detectable 4 days post‐infection (pi), reached its peak level about 10 days pi, and faded away rapidly in about 10 days thereafter.
Studies on the Experimental Cardiac Involvement and Cell‐Mediated Immunity in Germ‐Free Mice Sensitized with Killed Group A Streptococci
Authors: Iwaku Tateo
Published: November 2013

Electrocardiographic changes and cell‐mediated immunity in germ‐free mice sensitized with formalin‐killed group A streptococci were studied. In germ‐free mice sensitized with formalin‐killed group A streptococci, a marked prolongation of the PQ interval in electrocardiogram (ECG) was observed at the fourth and seventh week of the sensitization, and significant increases of [3H] thymidine uptake of lymphoid cells by stimulation with phytohaemagglutinin (PHA‐M), pokeweed mitogen (PWM) and streptococcal mucopeptide were observed at the second, third and sixth week of the sensitization. Moreover, in macrophage migration inhibition test, migration inhibition of macrophages by streptococcal mucopeptide antigen was seen at the third, fifth and sixth week, and by heat‐killed streptococci at the first and third week of the sensitization. In contrast, no electrocardiographic change and slight increases of [3H] thymidine uptake of lymphoid cells by stimulation with mitogens or streptococcal antigens were observed in non‐sensitized germ‐free mice and also in germ‐free mice sensitized with formalin‐killed staphylococci or treated with physiological saline.

In germ‐free mice sensitized with streptococci, their body weight did not increase and cyanosis on lips or extremities of the mice was noted throughout the experimental period.

Light microscopic histological examination could not reveal any changes that suggest damage of the myocardial tissue.
Inhibition by Potassium Ion of the Pregerminative Response to L‐Alanine of Unactivated Spores of Bacillus cereus T
Authors: Shibata Hirofumi , Takamatsu Hiroaki , Minami Masako , Tani Isamu
Published: November 2013

The effect of potassium ion on L‐alanine‐inosine‐induced germination of unactivated spores of Bacillus cereus T was studied. Unactivated spores germinated in 0.1 M sodium phosphate buffer (NaPB), but not 0.1 M potassium phosphate buffer (KPB), at pH 8.0 and at 30 C. Inhibition of germination was also observed on incubation of unactivated spores in NaPB containing potassium chloride. Previously it was demonstrated that germination of unactivated spores involves at least two steps, one induced by L‐alanine, and the other by inosine. Potassium ion seems to inhibit the response of the spores to inosine, because: (1) Spores that had been preincubated with L‐alanine in NaPB or KPB, germinated in NaPB but not KPB in the presence of inosine. (2) During germination in NaPB, incorporation of L‐[14C] alanine showed bimodal kinetics with a rapid first phase and a second continuous phase, but in KPB the second phase of incorporation did not occur.

The events occurring before germination of unactivated spores are discussed with reference to the initiation of germination.
Process of Consecutive Cell Divisions and Separations in a Regular Tetrads‐Forming Mutant of Micrococcus lysodeikticus (luteus)1
Authors: Monodane Toshihiko , Matsushima Yoshio , Kotani Shozo
Published: November 2013

A mutant MT of Micrococcus lysodeikticus (luteus) IFO 3333, whose minimum growing unit is not a single cell, but a tetrad unlike the wild‐type divides by binary fission of each monococcus, and then separates first into two daughter tetrads, second into four tetrads and third into eight tetrads. The three planes of either the cell division or the cell separation are equivalent to one another and oriented at right angles in three dimensions, respectively. The process of consecutive cell divisions and separations of the mutant tetrads was schematically illustrated.
Electron Microscopic Studies of Tumor Viruses
Authors: Miyamoto Kaneatsu , Sairenji Takeshi , Hinuma Yorio
Published: November 2013

Entry of Epstein‐Barr virus into human lymphoblastoid cells (Daudi cells) was studied by electron microscopy. At the site of viral attachment, two distinct interactions conducive to penetration of the virus occurred between the viral envelope and cell membrane, namely, (i) simultaneous dissolution of both the envelope and cell membrane, presumably resulting in passage of viral capsids into the cytoplasm and (ii) dissolution confined to the cell membrane with resulting penetration of enveloped virus. In the latter case envelope dissolution appears to occur subsequently in the cytoplasm with release of capsids. Fusion of the viral envelope with the cell membrane was not observed.

The capsids exhibited two distinct structural forms—one dense, the other translucent or light in appearance. The former disrupted near the cell membrane with release of viral cores into the cytoplasm whereas the light capsids containing dense cores migrated toward the nucleus and accumulated in the perinuclear region. Apparently the process of releasing deoxyribonucleic acid (DNA) from the light capsid is slowed down or prevented in Daudi cells. A hypothesis is presented concerning the manner in which these two types of capsids initiate infection.
New Plasmid‐Mediated Phosphorylation of Gentamicin C in Staphylococcus aureus
Authors: Kawabe Haruhide , Naganawa Hiroshi , Kondo Shinichi , Umezawa Hamao , Mitsuhashi Susumu
Published: November 2013

A clinical isolate of Staphylococcus aureus resistant to gentamicin, kanamycin and 3′,4′‐dideoxykanamycin B contained two enzymes capable of inactivating gentamicin, i.e., an aminoglycoside 2″‐phosphotransferase and aminoglycoside acetyltransferase.
Studies on the Pathogenicity of Human‐Origin Parainfluenza Virus in the Brain of Mice
Authors: Shimokata Kaoru
Published: November 2013

The pathogenicity of parainfluenza type 2 (croup‐associated) virus known to cause infections of the respiratory tract in the early life of man was studied in the brain of mice. One‐ to 4‐day‐old sucklings and 4‐week‐old mice were inoculated intracerebrally with the virus. The virus multiplied in sucklings, but not in adults. Most mice inoculated intracerebrally with the virus appeared healthy. Histological examination showed minimum inflammatory changes, although moderate hydrocephalus developed in three of twenty‐one sucklings by 6 weeks post‐infection. Immunofluorescent study in sucklings showed viral antigens in ependymal lining cells and choroid plexus epithelium during the first two weeks, and parenchymal cells for more than two months. Virus specific antibody response was observed in adults, but not in sucklings. One interesting finding was that viral antigens persisted in six out of 11 suckling brains for one to two months.
In Vitro Induction of Antibody‐Dependent Cytotoxic Macrophages by the Local Anesthetic Lidocaine
Authors: Kurisu Mariko , Yamazaki Masatoshi , Mizuno Den'ichi
Published: November 2013

Normal macrophages were activated to antibody‐dependent cytotoxic effector cells by in vitro treatment with the local anesthetic lidocaine. Experiments on the dose‐response and time course of the effect of lidocaine showed that incubation of normal macrophages with 10 mM lidocaine for 10 min at 28 C was enough for induction of antibody‐dependent cellular cytotoxicity. The activation by lidocaine was accompanied by enhanced phagocytosis of sheep red blood cells (SRBC) sensitized with anti‐SRBC antiserum, but not enhanced ingestion of polystyrene latex particles (PLP). These findings suggest that lidocaine, which has various effects on cell membranes, induces some perturbation of macrophage membranes, resulting in activation of Fc receptor functions in antibody‐dependent cytotoxicity and phagocytosis.
Effect of Intestinal Flora on Megaenteron of Mice
Authors: Itoh Kikuji , Maejima Kazuyoshi , Ueda Katsumoto , Fujiwara KÔSaku
Published: November 2013

CF#1 germfree (GF) and conventional (CV) mice as well as offspring of conventionalized GF (GF‐CV) mice were orally inoculated with Escherichia coli 0115a, c: K(B), a causative agent of megaenteron in mice. Although CV and GF mice showed no clinical signs and survived, all of the GF‐CV mice died with diarrhea by day 14 after inoculation. Thickened wall of the large intestine, microscopically showing proliferation of crypt type cells, was seen in GF and GF‐CV mice but not in CV mice. In addition, in GF‐CV mice, hemorrhage and severe erosion with marked inflammatory reactions were observed. While the inoculated E. coli could not colonize in CV mice, a level of 108 cells/g feces was maintained in GF mice from day 1 after inoculation to the end of examination (on day 28) and in GF‐CV mice from day 5 to the time of death. Newly prepared germfree (GF‐CV‐GF) mice obtained hysterectomy from GF‐CV mice showed a low sensitivity as comparable to that in GF mice. On the other hand, ex‐germfree mice produced by oral administration of feces of GF‐CV mice showed severe infection as comparable to that seen in GF‐CV mice. These results suggest that the intestinal flora may play roles both on protecting from the infection of pathogenic E. coli and on enhancing the infection.
Induction of C‐Type Virus in Cell Lines Derived from Calf Form Bovine Lymphosarcoma
Authors: Onuma Misao , Okada KŌSuke , Yamazaki Yooko , Fujinaga Kei , Fujimoto Yutaka , Mikami Takeshi
Published: November 2013

For attempt to detect an etiological agent, cultures from bovine lymphosarcoma cases (adult form (ALS), calf form (CLS), and thymic form (TLS)) were maintained in vitro for over a 18 month period. In two cultures from ALS, bovine leukemia virus (BLV) antigen was constantly detected. On the other hand, BLV antigen remained negative in cultures from two CLS and one TLS cases up to 40 passages. The RNA dependent DNA polymerase activities in these cultures were also negative.

Treatment of a culture from CLS (3178) originated from liver tumor with 5′‐iodo‐2′‐deoxyuridine (IdU) and dexamethasone (DXM) resulted in production of an agent serologically and morphologically similar to BLV and in alteration of cell morphology. No virus was detected in culture from TLS after treatment with IdU and DXM.
In Vitro Assay for Responsiveness of Lymphocytes to Transfer Factor by a New Leukocyte Migration Inhibitory Test
Authors: Miyagawa Yukiaki , Kawasaki Akiko , Komiyama Atsushi , Akabane Taro
Published: November 2013

Transfer factor (TF) causes nonimmune lymphocytes to produce leukocyte migration inhibitory factor (LMIF) in the presence of purified protein derivative (PPD). The activity of TF was measured by leukocyte migration inhibitory test (LMIT). The LMIT was a modification of the conventional agarose droplet method. To express the activity of LMIF quantitatively and simply, LMIF titer was introduced. The LMIF titer was obtained from the combination of two factors, LMIF dilution and cell migration diameter, and therefore this made the LMIT much more sensitive as compared to the conventional LMIT. The responsiveness of lymphocytes from acute lymphoblastic leukemia (ALL) and from cell‐mediated immunodeficiency in children to TF was assayed by LMIT. In ALL, the lymphocyte responsiveness was poor in relapse but improved with remission. The responsiveness was remarkably well in 3 patients with cell‐mediated immunodeficiency. This method appears useful for the in vitro evaluation of responsiveness of lymphocytes to TF.