bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of Host Deoxyribonucleic Acid Synthesis after Infection of Ultraviolet‐Irradiated Measles Virus
Authors: Minagawa Tomonori , Nakaya Chikako , Oguma Keizi , Iida Hiroo
Published: November 2013

Measles virus suppressed cellular DNA synthesis two hours after infection but did not inhibit RNA or protein syntheses. The suppression of DNA synthesis was not dependent on the cellular growth phase but was dependent on the multiplicity of infection. This activity of measles virus was inactivated neither by ultraviolet‐irradiation nor by heating at 56 C for two hours, but was lost after treatment with a convalescent antiserum of a measles patient or with proteolytic enzymes (pronase and trypsin).
Virulence of Rifampicin‐Resistant Mutants ofShigellaand EnteropathogenicEscherichia coliwith Special Reference to Their Cell Invasiveness
Authors: Osada Yasuaki , Une Tsutomu , Nakajo Masayuki , Ogawa Hidemasa
Published: November 2013

1. Changes in virulence ofShigellaandShigella‐like enteropathogenicEscherichia coli, which occurred during their acquiring resistance to rifampicin were examined. Virulence was evaluatedin vitroby their cell‐invasive activity, infectivity to the eye of guinea pigs, and LD50values in intraperitoneally infected mice. 2. The mutants obtainedin vivoand those obtainedin vitrosomewhat varied in cell invasiveness. Ability to produce keratoconjunctivitis in guinea pigs was lacking or diminished in all mutants though the parent strains constantly showed high abilities to invade cultured cells or produce kerato‐conjuctivitis in guinea pigs. 3. Mutations to resistant strains were not accompanied by any changes in LD50values in mice.
The Effect of Beta‐Lipoprotein on Antistreptolysin‐O Antibody Titers
Authors: Baba Toru , Suzuki Mitsuru , Sonozaki Hidekichi , Schröter Gerhard , Torisu Motomichi
Published: November 2013

The relationship between antistreptolysin‐O (ASLO) titers and β‐lipoprotein were examined. The following results were obtained. 1. Fifty‐three children with unmeasurably low ASLO titers had variable β‐lipoprotein concentrations which were within normal ranges. 2. Streptococcal infections in the children correlated well with the elevation of ASLO titers, but β‐lipoprotein concentration in these sera did not show any correlation with the behavior of ASLO titers. 3. From the Sephadex G‐200 gel filtration chromatographical study, no ASLO activities were found in the fraction containing β‐lipoprotein except in some special cases that had the IgM ASLO antibody. 4. The inhibitory effect of β‐lipoprotein upon hemolysis by streptolysin‐O was incomplete and its effect decreased with a prolonged incubation time. 5. The partially purified β‐lipoprotein had no effect upon the hemolytic activity of streptolysin‐O, when the ASLO titers were measured by the method of 100% inhibition of hemolysis. From these observations, it was concluded that the ASLO antibody titers were truly related to streptococcal infection and not influenced by the presence of β‐lipoprotein if the ASLO titers were measured by the 100% inhibition of hemolysis.
A Lysin(s) in Lysates of Clostridium botulinum A190 Induced by Ultraviolet Ray or Mitomycin C
Authors: Mitsui Noriko , Kiritani Kazuyoshi , Nishida Shoki
Published: November 2013

When cells of the Clostridium botulinum A190 strain were subjected to treatment with mitomycin C or to irradiation with ultraviolet ray and then grown at 37 C, an induced lytic agent(s) was produced in the resultant lysate. Production of the lytic agent in the induced bacterial culture was inhibited in the presence of chloramphenicol. A crude preparation of the phage‐free lytic agent readily lysed freeze‐thawed or acetone‐treated cells of C. botulinum B‐NIH19, and heated or chloroform‐treated cells to a lesser extent at the optimal pH of 6.8. The lytic agent was stable at or below 37 C, but unstable above 37 C. A crude preparation of the lytic agent did not exhibit a detectable effect on the viability of vegetative cells.
Numerical Taxonomy of Clostridium botulinum and Clostridium sporogenes Strains, and Their Susceptibilities to Induced Lysins and to Mitomycin C
Authors: Kiritani Kazuyoshi , Mitsui Noriko , Nakamura Shinichi , Nishida Shoki
Published: November 2013

An attempt to classify fourteen strains of Clostridium sporogenes and thirty strains of Clostridium botulinum types A, B, C, D, E, and F by numerical taxonomy was made. All proteolytic strains of C. botulinum types A, B, and F and C. sporogenes were classified in the phenon I at a level of 86% S‐value. In the phenon I, these strains coexisted in a mixed‐up fashion, irrespective of conventional species or type of bacteria. A concentrated lysin solution, which was prepared from an induced lysate of a C. botulinum A190 culture, lysed vegetative cells of all proteolytic C. botulinum and C. sporogenes strains classified in the phenon I, but did not lyse the cells of the other Clostridia. However, cells of nonproteolytic C. botulinum F‐OSU and four out of five strains of Clostridium tetani were lysed by the concentrated lysin to a limited extent. The phenon II contained all nonproteolytic strains of C. botulinum B, C, D, E, and F, which were mutually linked with S values of more than 85%. Strains of Clostridium histolyticum used as a reference group formed the phenon III. Any strain grouped in one phenon was differentiated from strains grouped in the other phenons by low S‐values, that were less than 77%. A striking difference in the number of strains susceptible to mitomycin C was demonstrated between C. botulinum and C. sporogenes. In all thirty‐five cultures, except four substrains of C. botulinum B‐NIH, of proteolytic and nonproteolytic C. botulinum, bacterial lysis was consistently induced by the treatment of 1 μg per ml of mitomycin C, while only four out of thirty‐seven C. sporogenes strains were as sensitive to mitomycin C as C. botulinum. Toxigenic substrains of C. botulinum B‐NIH, NIH15 and NIH19, were sensitive to mitomycin C, but nontoxigenic strains, NIH5, and NIH‐NP, were not.
Development of Resistance to and Dependence on Streptomycin in Mycoplasma pneumoniae In Vitro
Authors: Kubota Hideo
Published: November 2013

Mycoplasma pneumoniae developed high degrees of resistance to streptomycin and dependence on streptomycin after serial subculture in broth media containing streptomycin. The streptomycin‐dependent organisms failed to grow, as indicated by a color change of the medium, in the absence of streptomycin but grew in limited concentrations of streptomycin in broth media. The minimum growth‐inhibitory concentration of streptomycin for the parent sensitive strain was very close to the minimum concentration of streptomycin which permitted the growth of the streptomycin‐dependent organism. The broth cultures showing streptomycin dependence were shown to consist of streptomycin‐dependent organisms or a mixture of streptomycin‐dependent and ‐resistant organisms, as tested on agar media containing varying concentrations of streptomycin.
Fate of Tobacco Mosaic Virus after Entering the Host Cell
Authors: Hayashi Takaharu
Published: November 2013

Tobacco leaves were inoculated with tobacco mosaic virus labeled with 32P or 35S. After various intervals, extracts of the leaves were prepared. In extracts from leaves infected for 5 to 360 min, about 40 to 60% of the virus retained on leaves was recovered in the pellet of the homogenate centrifuged at 12 000 × g. The virus associated with the 12 000 × g pellet was dissociable by treatment with pancreatic RNase, alkali or sodium dodecyl sulfate (SDS). The parental virus extracted by SDS from the pellet at 12 000 × g had a large amount of partially uncoated virus possessing naked RNA. Analysis by density gradient centrifugation suggested that, in addition to partially uncoated virus, some fragmented RNA was also associated with the 12 000 × g pellet. This fragmented RNA seemed to be derived from partially uncoated virus. Density gradient analysis of SDS extracts from the 12 000 × g pellet suggested that some of the virus underwent uncoating at the internal regions of the virus particle.
Antiviral Activity of Scillarenin, a Plant Bufadienolide
Authors: Sato Norio , Muro Toshio
Published: November 2013

Antiviral activity of scillarenin, a plant bufadienolide, and its mode of action were studied. Scillarenin selectively inhibited replication of picornaviruses, in particular, rhinovirus. At concentrations inhibiting rhinovirus replication, scillarenin had no effect on the synthesis of macromolecules in host cells. The action of scillarenin was not direct inactivation of virus. Scillarenin could inhibit virus replication when given at the latter half of latent period, although this treatment did not interfere with the normal level of viral RNA synthesis. The RNA synthesized in the presence of this compound showed the same sedimentation coefficient as normal viral RNA but possessed a reduced infectivity.
In Vitro Reconstitution of Anti‐Sheep Erythrocyte Antibody Response of T Cell‐Depleted Spleen Cells by Allogeneic T Cells or by Factors Derived from Them
Authors: Tadakuma Takushi , Mitsuma Takashi , Ishikawa Hiromichi , Saito Kazuhisa
Published: November 2013

Restoration of the impaired antibody response to sheep erythrocytes (SRBC) in cultures of mouse spleen cells, which were deprived of thymus‐derived lymphocytes (T cells) by treatment with anti‐mouse brain‐associated θ (BAθ) antiserum and complement, was studied by adding a small portion of syngeneic or allogeneic normal spleen cells in vitro. Allogeneic spleen cells had a far greater effect than syngeneic spleen cells on the restoration, as far as the normal spleen cells added were able to recognize the alloantigens on the anti‐BAθ serum‐treated spleen cells (bone marrow‐derived lymphocytes). Treatment of the allogeneic spleen cells with mitomycin C did not affect their activity in the restoration of the impaired antibody response. The possibility that the role of T cells in the antibody response to SRBC may be replaced by a nonspecific mediator derived from T cells reacting with allogeneic cells was proven by the finding that supernatant of the mixed allogeneic spleen cell cultures restored the impaired anti‐SRBC antibody response of the T cell‐depleted spleen cells. The effect of such culture supernatant on the restoration of the antibody response was greatest when it was added to the T cell‐depleted spleen cell cultures one day after cultivation with SRBC, suggesting that the effectiveness may result from triggering of the proliferation and differentiation of antibody‐forming cell precursors, which have already reacted with the antigen, to antibody‐forming cells.
Induction of Phage Production in the Lysogenic Escherichia coli by Hydroxyurea
Authors: Shimada Kazunori , Shibata Yosaburo , Takagi Yasuyuki
Published: November 2013

1) Hydroxyurea, a reversible DNA synthesis inhibitor, was used to study the mechanism of prophage λ induction in Escherichia coli K12. Induction of prophage was judged on two criteria: increase of phage‐producing cells and loss of colony‐forming ability of the cells. 2) Hydroxyurea induced an increase of phage‐producing cells only in lysogenic strains known to be inducible with ultraviolet irradiation for prophage development and not in strains such as E. coli K12 (λind–) or E. coli K12 recA (λ+). 3) When protein synthesis was inhibited, hydroxyurea did not increase phage‐producing cells of lysogenic strains; it showed a bacteriocidal effect on lysogenic recA+ strains, but not on nonlysogenic strains. 4) The sensitivity of E. coli K12 recA to hydroxyurea was independent of whether or not the cells were lysogenic. 5) From the results it is suggested that certain steps leading to loss of colony‐forming ability (i.e. prophage induction) do not require de novo protein synthesis but require the presence of the host recA+ gene.
Skin Reaction and Macrophage Migration Inhibition Tests for Polysaccharides from Aspergillus fumigatus and Candida albicans
Authors: Suzuki Masuko , Hayashi Yoshiro
Published: November 2013

Guinea pigs sensitized with purified galactomannan from Aspergillus fumigatus and mannan from Candida albicans, each containing negligible quantities of nitrogen, were examined for their immunological responses against the corresponding polysaccharides with respect to the delayed‐type skin reaction and the macrophage migration inhibition phenomenon. In both cases, the delayed‐type skin reaction test and the macrophage migration inhibition test showed positive results. The reactivity was stronger in animals sensitized with polysaccharides in Freund's complete adjuvant than those sensitized with the same polysaccharides in Freund's incomplete adjuvant. Polysaccharides chemically modified by partial acid degradation or by periodate oxidation were found to be completely incapable of eliciting such immune responses. These results are also discussed in relation to the antigenic determinant of the polysaccharides in such immune responses and the precipitin reaction previously observed by us and other investigators.
The Effect of 2,4‐Dinitrophenol on the Growth of Bordetella pertussis in Chick Tracheal Organ Culture
Authors: Iida Tsuyoshi , Ajiki Yoko
Published: November 2013

Among various metabolic inhibitors tested, only 2, 4‐dinitrophenol inhibited the growth of Bordetella pertussis in chick tracheal organ culture at concentrations nontoxic both for bacterial organisms and for ciliary motility of the tracheal fragments. Although this effect of 2, 4‐dinitrophenol was reversible in its early stage, longer treatment with this inhibitor resulted in an irreversible inhibition of bacterial growth due to secondary damage of the tracheal fragments. From these observations, it was postulated that the energy required for bacterial growth might be derived from cellular metabolism sensitive to inhibition with 2, 4‐dinitrophenol.
Ribonucleic Acid‐Dependent Ribonucleic Acid Polymerase in the Immune Response
Authors: Saito Kazuko , Mitsuhashi Susumu
Published: November 2013

Ribonucleic acid (RNA)‐dependent RNA polymerase activity was demonstrated in the microsomal and ribosomal fraction from the spleen cells of immunized mice. The enzyme activity was solubilized by Triton X‐100 from the fraction and partially purified by Biogel A 1.5 M column chromatography. The RNA‐dependent RNA polymerase activity was eluted in a single peak from the column. High activity was demonstrated with an RNA preparation (iRNA) as template made from the spleens of immunized mice but very low activity was found with an nRNA preparation made from the spleens of normal mice. Incorporation of 3H‐UTP markedly decreased in the presence of RNase but not in the presence of DNase. DNA preparations made from the spleens of immunized mice were inactive as template for this enzyme. The iRNA preparation was fractionated by sucrose density gradient centrifugation. A fraction corresponding to 12–13 S was most active as a template. It was followed by a fraction corresponding to 6–7 S. Sucrose gradient analysis of the 3H‐UTP‐labeled product was attempted. Some properties of this enzyme are described.
Ultrastructure and Chemical Analysis of the Cell Wall of Pythium debaryanum1
Authors: Yamada Michihisa , Miyazaki Toshio
Published: November 2013

The ultrastructure and component polysaccharides of the cell wall of Pythium debaryanum IFO‐5919 were investigated. From results obtained by means of acid, alkali, Schweitzer reagent and β‐1, 3‐glucanase treatments and electron microscopy, it was concluded that 1) the acid‐extracted fraction was a 1,3‐linked branched glucan, 2) the alkali‐extracted fraction was a mixture of 1,3‐, 1,6‐, and 1,3,6‐linked highly branched two glucans, 3) the Schweitzer reagent‐extracted fraction was a β‐1, 4‐linked glucan, 4) the cell wall was constructed from two types of cullulosic microfibrils, as a frame and as a finer network, and amorphous β‐1, 3‐glucan including β‐1, 6‐linkage, 5) cellulosic microfibrils were covered by matrix material consisting of a mixture of amorphous β‐1, 3‐linked and β‐1, 6‐linked branching glucans.
Mitogenic Activity of the Cell Walls of Mycobacteria, Nocardia, Corynebacteria and Anaerobic Coryneforms
Authors: Azuma Ichiro , Taniyama Tadayoshi , Sugimura Kazuhisa , Aladin Aminkham A. , Yamamura Yuichi
Published: November 2013

The mitogenic activity of the cell walls prepared from Mycobacterium bovis BCG, Nocardia rubra, Corynebacterium diphtheriae PW8, and four species of Propionibacterium, Corynebacterium parvum ATCC 11829, Propionibacterium acnes C7, Propionibacterium granulosum ATCC 25564 and Propionibacterium avidum ATCC 25577, were investigated. These cell walls were active as mitogens on normal spleen cells, anti‐θ sera‐treated spleen cells, macrophage‐depleted spleen cells of C57BL/6J mice and cortisone‐treated thymocytes of C57BL/6J mice. It was also shown that these cell walls were mitogenic on spleen cells and macrophage‐depleted spleen cells of congenitally athymic (nude) mice. The above results suggest that the cell walls investigated in this study act as mitogens on both thymus‐derived lymphocytes (T‐cells) and bone marrow‐derived lymphocytes (B‐cells).
Alteration of the In Vitro Host Range of Rabies Virus after Serial Chick Embryo Cell Passage Using Alkaline Maintenance Medium
Authors: Hashimoto Mitsue , Yoshino Kamesaburo
Published: November 2013

HEP Flury strain of rabies virus maintained by 7‐day chicken egg passage (parent line) and the same strain serially passaged in primary chick embryo (CE) cells using alkaline maintenance medium (AM line) were inoculated to cells of various species. Growth was negative in primary mouse embryo, L and HeLa cells, and positive in primary hamster kidney and BHK21 cells with both lines. An all‐or‐none difference between the two lines was observed in primary monkey kidney and Vero cells. The parent line did not multiply in these monkey cells, whereas the AM line grew to high titers. In the case of Vero cells a unique cytopathic effect (CPE) was induced by the AM line. After five consecutive passages in Vero cells, the CPE‐inducing agent was identified as rabies virus by a neutralization test. It was infective to intracerebrally inoculated suckling mice but not to adult mice, and its Vero cell‐infective titer determined by CPE induction was about 1 log lower than the baby mouse‐infective and CE plaque‐forming titers. In contrast to the AM line, HEP Flury strain receiving 150 CE cell passages under neutral maintenance medium and three other strains receiving similar CE cell passages all failed to grow in Vero cells.
Isolation and Biological Characterization of a Measles Virus‐Like Agent from the Brain of an Autopsied Case of Subacute Sclerosing Panencephalitis (SSPE)
Authors: Makino Satoshi , Sasaki Keiko , Nakagawa Masaharu , Saito Mitsuji , Shinohara Yukito , Gotoh Fumio , Okabe Takashi
Published: November 2013

Isolation of a cytopathic agent causing formation of syncytial giant cells in co‐cultivated Vero cells from the brain of an autopsied case of subacute sclerosing panencephalitis (SSPE) is reported. The syncytia usually autolyzed from the center after growing to 1 to 2 mm in diameter and then detached from the culture vessels, and finally made macroscopically recognizable round plaques on the monolayer under liquid overlay. The agent was identified serologically as an agent related to measles virus, by both immunofluorescent tests and plaque reduction tests using anti‐measles sera. However, the infected cells did not produce either virions or hemagglutinin, and failed to show hemadsorption and hemolysis of African green monkey red cells even after the 55th passage through Vero cells. Newborn mice, adult mice and hamsters showed neurologic signs after intracerebral inoculations of the infected cells, and most of them died from acute encephalitis. Guinea pigs were unsusceptible. From the brain of the animals with neurologic signs, a similar agent to the inoculated one was recovered.

The new isolate appears to be a strain closely related to measles virus on the basis of serology, and was designated as SSPE‐“Kitaken‐1” strain.
Studies on the Neutralization of Herpes Simplex Virus
Authors: Yoshino Kamesaburo , Hashimoto Mitsue , Shinkai Kenkichi
Published: November 2013

Early and late IgG of rabbits immunized with herpes virus showed, respectively, 8‐fold and 2‐fold enhancement of neutralization endpoint in the presence of complement (C). Kinetic curve experiments employing an appropriate amount of virus revealed that both neutralization and sensitization followed first‐order reaction, and each IgG possessed a certain range of concentration where neutralization was negligible while sensitization was marked. Dose responses of neutralization and sensitization velocities demonstrated that the C enhancement of late IgG was about 7‐fold and that of early IgG more than 20‐fold. These facts suggested that the IgGs contained two different entities of complement‐requiring (CRN) and non‐requiring neutralizing (N) antibodies at different proportions, only the former being responsible for sensitization. The different CRN: N ratios obtained by the endpoint and kinetic methods may mean either that the two antibodies differ in avidity for the virus or that the number of critical sites per virion for CRN antibody is greater than that for N antibody. In this interpretation, sensitization by CRN antibody as well as neutralization by N antibody is thought to result from attachment of a single antibody molecule to the viral critical site. Alternative explanations, ascribing the mechanism of neutralization to steric hindrance of critical sites or to multiple hit of those sites by antibody, were denied by analyses of the present data.
Microbial Adjuvant and Autoimmunity
Authors: Nakashima Izumi , Yokochi Takashi , Kato Nobuo , Asai Junpei
Published: November 2013

The target organs of mice immunized with the respective syngeneic tissue extracts together with the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) as a powerful adjuvant were examined for production of lesions. In 15 out of 24 mice injected three times or more with syngeneic eyeball extracts and CPS‐K adjuvant at intervals approximately 30 days, severe eyeball lesions developed in which the normal structure was almost completely lost. A large part of the eyeball tissue of these mice was replaced by infiltration with cells such as lymphocytes, plasma cells and other mononuclear cells and by connective tissue. No definite eye lesions developed in mice injected with CPS‐K alone, eyeball extracts alone or eyeball extracts emulsified in complete Freund's adjuvant (CFA). In all of mice injected four times with thyroid gland extracts and CPS‐K at intervals of approximately 30 days, definite thyroid gland lesions were produced. In three out of five mice of this group, the thyroid lesions were so severe that the normal thyroid follicular structure was almost completely lost, and a large part of the thyroid gland was replaced by infiltration with lymphocytes, plasma cells and other mononuclear cells and in part by connective tissue. In only one out of five mice injected with thyroid gland extracts emulsified in CFA, definite but milder thyroid gland lesions developed. No definite thyroid lesions developed in the remaining four mice of this group and also in any of the mice injected with thyroid gland extracts alone or CPS‐K alone. Repeated injections of lymphoid tissue extracts and CPS‐K also induced pathological changes in the spleen and lymph nodes, although less marked than those in the cases of the eyes and thyroid gland. The most remarkable change was a decrease in numbers of small lymphocytes at the areas surrounding the central arterioles in the white pulp of the spleen and the post‐capillary venules in the cortex of the lymph nodes. From these results it has been concluded that our system can provide new and useful models for autoimmune diseases in man.
Studies on the Pathogenicity ofYersinia enterocolitica
Authors: Une Tsutomu
Published: November 2013

Analysis of the pathogenicity ofYersinia enterocoliticawas performed with an experimental model successfully produced in rabbits by intraduodenal inoculation with strains isolated from various sources.

Pathogenic strains easily penetrated the epithelial linings of the intestinal mucous membrane into the target reticuloendothelial tissues of the intestine, such as the lamina propria and lymph follicles, where they multiplied within mononuclear cells and produced granuloma.

Granuloma, in severe infections, underwent necrobiosis and sometimes progressed to ulceration accompanied by colony formation of the organisms.

In mild infections, granulomatous lesions were localized in lymph follicles and never progressed to ulceration.

Nonpathogenic strains were rapidly excreted without penetration of epithelial linings.

Y. enterocoliticashould be within the category of invasion type enteropathogenic bacteria such asShigellaandSalmonella. Pathogenic behavior ofY. enterocoliticais discussed in comparison with that ofShigellaandSalmonella.
Intracellular Development of Membrane Protein of Influenza Virus
Authors: Maeno Koichiro , Yoshii Saiji , Yoshida Tetsuya , Iinuma Masao , Kawamoto Yasuko , Matsumoto Toshisada
Published: November 2013

The intracellular development of membrane protein (MP) of influenza A virus was investigated by immunofluorescent staining. Monospecific antiserum was prepared by immunizing rabbits with MP eluted from SDS‐polyacrylamide gels of SDS‐disrupted NWS virions. In the productive infection in clone 1‐5C‐4 cells, MP antigen was first detected over the whole cell at 4 hr after infection, concomitantly with the appearance of hemagglutinin (HA) antigen in the cytoplasm, and bright nuclear fluorescence was then observed. Nucleoprotein (NP) antigen was detected in the nucleus prior to the appearance of fluorescence of MP antigen and thereafter the cytoplasmic fluorescence developed. Late in infection, all of these three antigens were observed predominantly in the cytoplasm with stronger fluorescence at the cell surface. Essentially similar findings were obtained in the abortive infections in L cells and BHK cells.

The above results suggest that the membrane protein of influenza A virus is present in the nucleus as well as in the cytoplasm of infected cells.
Regulation of Anti‐Hapten Antibody Response by Chemically Modified Carrier Antigen Preferentially Provoking Delayed‐Type Hypersensitivity (DTH)
Authors: Machida Atsuhiko , Kumazawa Yoshio , Mizunoe Kimifusa
Published: November 2013

Experiments were carried out to determine whether or not the cell populations involved in DTH and in the suppression of antibody response are identical. The effects of four treatments, i.e., adult thymectomy (ATx), X‐irradiation, anti‐mouse thymocyte serum (ATS) and hydrocortisone (HC) on the induction of DTH and on the carrier‐specific suppression of antibody response were observed in mice immunized with chemically modified antigen, dodecanoyl‐BSA (d‐BSA), emulsified with complete Freund's adjuvant (CFA), with the following results: 1) DTH induced by immunization with D‐BSA remained constant in adult thymectomized mice, whereas the suppression of antibody response was not inducible in these animals. 2) Injection of low doses of ATS caused the depression of DTH in mice primed with D‐BSA, but did not affect the suppressive activities of their spleen cells. 3) Sublethal X‐irradiation 1 week prior to D‐BSA priming inhibited the generation of suppressor cells but did not affect the generation of cells mediating DTH. The suppressive effect was also abrogated by sublethal X‐irradiation given 2 days after immunization with DNP‐BSA (14 days after priming with D‐BSA). 4) The treatment of animals with HC 2 days before the footpad challenge or immunization with DNP‐BSA depressed the ability of animals to induce both DTH and the suppression of antibody response. However, the latter was more sensitive to HC than the former.

In addition to these results, it was also found that D‐BSA‐primed spleen cells were capable of suppressing anti‐DNP response, but not of inducing DTH‐reactivity upon transfer to recipient mice. These results suggest that DTH‐reactivity and the carrier‐specific suppression of anti‐hapten antibody response induced by injection of D‐BSA are mediated by different cell populations.
Reduction of Population Levels of Some Indigenous Bacteria by Lactobacilli in the Gastrointestinal Tract of Gnotobiotic Rats
Authors: Watanabe Tsugio , Morotomi Masami , Kawai Yasuo , Mutai Masahiko
Published: November 2013

Effects of each of three indigenous Lactobacillus groups on other bacterial populations were separately investigated in gnotobiotic rats. In the wall of the non‐glandular part of the stomach, contents of the stomach and contents of the upper part of the small intestine, some pre‐associated indigenous bacteria were reduced to conventional population levels by introducing three groups of lactobacilli: Group I (Lactobacillus acidophilus and related strains), Group II (L. fermentum) and the groups mixed. However, no obvious reduction in cell numbers of the pre‐associated bacteria occurred in the case of Group III (L. murini).
Macrophage Dependency of T‐lymphocyte Mitogenesis by Nocardia rubra Cell‐Wall Skeleton
Authors: Sugimura Kazuhisa , Uemiya Masanao , Azuma Ichiro , Yamawaki Mikio , Yamamura Yuichi
Published: November 2013

The mitogenic activity of the cell‐wall skeleton (CWS) of Nocardia rubra on purified splenic T‐cells (thymus‐derived lymphocytes) was investigated. N. rubra CWS showed remarkable mitogenic activity on normal spleen cells of C57BL/6J mice at concentrations ranging from 10 to 100 μg per milliliter of culture medium, while, on purified splenic T‐cells, N. rubra CWS did not act as a mitogen at any concentration. However, mitogenic activity of N. rubra CWS on T‐cells was restored if purified splenic T‐cells were reconstituted with X‐irradiated peritoneal exudate cells (macrophages). The above results suggest the necessity of macrophages for T‐lymphocyte activation by N. rubra CWS as well as PHA‐P or Con A.
Mechanism of Defective Lysogenization by Phage P1 in a lon− Mutant of Escherichia coli K‐12
Authors: Takano Toshiya
Published: November 2013

Phage P1 cannot lysogenize a lon– mutant of Escherichia coli K‐12, which is defective in the regulation of cellular division cycle to result in snake formation (14). P1 mutants, called P1pla, can lysogenize the lon– host. These mutations have been classified into two complementation groups: one is cis‐dominant; the other is trans‐dominant. A temperature‐sensitive lon– mutant was isolated, which exhibited the lon– phenotype at 42 C but not at 33 C. A temperature‐shift experiment of the P1‐lysogenic derivative of the lon– ts mutant showed lysis of the culture and induction of the phage production. It is proposed that P1 plasmid may be under a certain regulatory circuit of the division cycle of the host bacterium by indirectly regulating the production of P1 immune repressor, or alternatively by directly derepressing the functions of P1 prophage.
The Effect of the DNA‐Suppressing Factor (DSF) on Host DNA Synthesis in Synchronized Cell Cultures
Authors: Minagawa Tomonori , Fujii Nobuhiro , Yamamoto Tomoko K. , Iida Hiroo
Published: November 2013

Purified host DNA‐suppressing factor (DSF) produced into culture fluid of HeLa C‐9 cells infected with measles virus inhibited cellular DNA synthesis in HeLa cells. When purified DSF was added into cultures of synchronous HeLa cells at the early G1‐phase, cellular DNA synthesis was irreversibly inhibited. However, DSF did not affect the stability of native double‐stranded DNA nor the chain‐elongation of single‐stranded DNA in cells of the S‐phase.
Improved Reversed Passive Hemagglutination for Simple and Rapid Detection of Staphylococcal Enterotoxins A~E in Food
Authors: Yamada Sumio , Igarashi Hideo , Terayama Takeshi
Published: November 2013

Detection and identification of staphylococcal enterotoxins in food or culture filtrates were performed using the reversed passive hemagglutination (RPHA) technique, with formalized sheep red blood cells (FSRBC) sensitized with immunoglobulins of anti‐A, B, C, D, and E rabbit hyperimmune sera fractionated by affinity chromatography. The FSRBC sensitized with anti‐A~E immunoglobulins showed a high level of reactivity and specificity in RPHA, against homologous types of purified enterotoxins and culture filtrates of toxin‐producing strains. No non‐specific reactions with various ingredients in foods nor cross‐reactions among enterotoxin types were observed. The minimum amount of enterotoxins in foods detected by RPHA was calculated to be 0.01 μg/g without concentration, and the recovery rate of experimentally added toxins was calculated to be about 80%.

Under routine laboratory practice, detection and identification of enterotoxins from incriminated foods of five food poisoning outbreaks were performed by RPHA within 3 hr after reception of the specimens. Among them, three were determined to be enterotoxin A food poisoning, one to be toxin C and the rest to be intoxication of A and D. The concentration of the toxins was between 0.014 and 3.65 μg per gram of food.
Persistent Infection with Mouse Hepatitis Virus of Low Virulence in Nude Mice
Authors: Tamura Teruko , Taguchi Fumihiro , Ueda Katsumoto , Fujiwara KÔSaku
Published: November 2013

A persisting type of infection with wasting syndrome was established in congenitally athymic nude mice after intraperitoneal inoculation with a mouse hepatitis virus which was not fully pathogenic for heterozygous haired littermates. From the liver, spleen, lymph nodes, and brain of most infected nude mice, the virus was detected at high titers during a period from 6 to 35 days postinfection, occurrence of degenerative and necrotic lesions being correlated with virus titers in these organs. The titer of serum neutralizing antibody remained undetectable or very low in most diseased nude mice, whereas some animals resisting the infection could produce antibody at a later stage. In heterozygous haired mice, some lesions were detectable at a very early stage of infection in the spleen and liver, but they seemed to disappear with a marked elevation of the neutralizing antibody titer. Nude mice were able to resist the virus infection when they had previously received transfer of thymocytes from weanling heterozygous littermates.
Immunological Studies of Heat‐Labile Virus Inhibitors
Authors: Kitamura Takashi , Tanaka Yukie , Ogata Momoko
Published: November 2013

Heat‐labile virus inhibitor (HLI) in normal sera of various mammalian species capable of neutralizing variola (VRV) and Newcastle disease viruses (NDV) was studied immunologically. After sucrose density gradient centrifugation of guinea pig serum, the HLI activity against VRV and that against NDV were both demonstrated in the same region sedimenting fastor than IgM. Absorption with partially purified VRV or NDV removed the HLI activity on the homologous virus but not that on the other. Prior saturation of virions with specific antibody blocked the absorption of HLI, suggesting a specific competition for binding site(s) between specific antibody and HLI.

The HLI level against variola virus was checked in connection with immunization with vaccinia virus. In human primary vaccination, the HLI level rose sharply within 4 weeks after vaccination, turning to decline gradually to settle at a level higher than that of the conventional neutralizing antibody (NA). In cases of human revaccination, a sharp rise of HLI started 4 days after vaccination and reached the highest level within 7 days, preceding the rise of conventional NA level which occurred about 3 days later. Three rabbits with negative HLI activity prior to vaccinia immunization obtained an HLI activity within 2 weeks, which showed a sharp rise up to 6–8 weeks. One rabbit with a positive prior HLI activity also showed a sharp rise of the HLI activity after immunization. In all rabbits the final HLI level was identical with that of conventional NA.

Groups of guinea pigs were immunized with either VRV or NDV. Rises of the HLI level after immunization were observed in all animals, the activity being restricted to the homologous virus used in the immunization. Complement‐requiring NA was detected during the course of immunization but its behavior was different from that of HLI.

The above observations were interpreted to suggest a ubiquitous presence of HLI as a specific reactive agent and its role at an earliest stage of immune response.
Complement‐Dependent Cytotoxicity of Sera Obtained from Subacute Sclerosing Panencephalitis Patients
Authors: Kuwajima Shigeru , Minagawa Tomonori , Fujii Nobuhiro , Iida Hiroo
Published: November 2013

Human lymphoid cells (NC‐37) persistently infected with either measles virus (Schwarz and TYCSA strains) or subacute sclerosing panencephalitis (SSPE) virus (Halle and Mantooth strains) were destroyed in the presence of complement by anti‐measles sera as well as by sera from SSPE patients. The cytotoxic activity was demonstrated in both IgG and IgM fractions of measles convalescent sera, but only in IgG fraction of SSPE sera. Measles convalescent sera completely lost the cytotoxic activity to all the cell lines, when absorbed with any one of the cell lines, indicating that the viral surface antigens of these cell lines infected with measles or SSPE virus are identical. On the other hand, the cytotoxic activity of SSPE sera could not be readily absorbed with these cells. Thus, the affinity of SSPE sera for the viral surface antigens might be lower than that of measles convalescent sera.
Studies on the Neutralization of Herpes Simplex Virus
Authors: Yoshino Kamesaburo , Isono Noriko
Published: November 2013

Early and late sera of rabbits immunized with herpes simplex virus were fractionated into IgG and IgM, and the minimal concentration of complement (C) required for full enhancement of neutralizing activity was determined for each by the plaque reduction method. In tests employing simultaneous mixing of virus, antibody and C, C‐requiring neutralizing (CRN) antibody in IgM required 2–8 times more C than that in IgG. When virus‐antibody mixtures were incubated at 0 C overnight before addition of C, a marked enhancement of CRN endopoint especially of late IgG and IgM was exhibited, in contrast to materially unchanged titers of the ordinary neutralizing antibody. This result suggested an abundance of slow‐reacting CRN‐virus complexes. The CRN antibody so detected required about 4 times more C than that detectable by the usual test in the case of late IgG and IgM. When virus sensitized with late IgG at 0 C overnight was further incubated at 37 C for 1 hr, the C requirement changed but slightly without showing any more increase of the endpoint, whereas sensitization at 0 C for 2 to 3 days further increased the CRN antibody endpoint but the C requirement was equal to that after 1 day's sensitization at 0 C. Based on these and earlier findings, a hypothesis is proposed that binding of a single antibody molecule with virus may cause a series of changes of the virus particle or part of those changes depending on the nature of antibody and on the sensitization condition, and C added to such complexes at an appropriate stage of the changes can accelerate the procession of the changes leading eventually to inactivation.
Nature of Antigenic Determinant of Serovar‐Specific Antigen of Leptospira interrogans Serovar hebdomadis
Authors: Adachi Yoshikazu , Yanagawa Ryo
Published: November 2013

The nondialyzable delipidized serovar‐specific main antigen (NDTM antigen) of Leptospira interrogans serovar hebdomadis strain Hebdomadis (a variant which can grow in a synthetic medium) showed a strong inhibition of the complement fixation between the serovar‐specific main (TM) antigen of this strain and the homologous antiserum. The inhibitory effect of the NDTM antigen was completely lost by treating the antigen with proteolytic enzymes, and the fractions of TM antigen containing amino sugar, neutral sugars, and lipids did not show any inhibition of complement fixation, indicating that the antigenic determinant of this strain is related to proteins. NDTM antigen contained more hydrophilic amino acids than hydrophobic amino acids, whereas TM antigen contained more hydrophobic amino acids than hydrophilic amino acids. The amino acid compositions of NDTM antigens of hebdomadis strain Hebdomadis (variant) and kremastos strain Kyoto, which belonged to the same serogroup, were considerably similar. Difference was found in the amounts of methionine, arginine, lysine and glutamic acid.
Modification of Immune Response in Nude Mice Infected with Mouse Hepatitis Virus
Authors: Tamura Teruko , Machii Kenji , Ueda Katsumoto , Fujiwara KÔSaku
Published: November 2013

In nude mice experimentally infected with mouse hepatitis virus (MHV), the numbers of early and later plaque forming cells (PFC) to sheep red blood cells (SRBC) generated in the spleen were 7 to 20 times and 2 to 163 times, respectively, greater than those in non‐infected nude mice, when SRBC were given at day 0 to day 21 postinfection. Splenic theta‐positive lymphocytes in infected nude mice were shown to increase only at day 10 or more postinfection. PFC response to bacterial lipopolysaccharide, a T cell‐independent antigen, was not modified in MHV‐infected nude mice.
Murine Model for Immunoprophylaxis of Cytomegalovirus Infection
Authors: Minamishima Yoichi , Eizuru Yoshito , Yoshida Aichi , Fukunishi Ryo
Published: November 2013

Murine cytomegalovirus was utilized as a model for human cytomeglovirus, which had no experimental animal, to study immunoprophylaxis of the cytomegalovirus infections.

Murine cytomegalovirus (MCMV) serially propagated in mouse embryonic fibroblasts had lost pathogenicity for weanling mice including neonatally thymectomized mice.

The cell culture‐adapted MCMV was effective as a “live, attenuated virus vaccine” against challenge by virulent, mouse‐passaged MCMV.

The immunization via intraperitoneal route protected mice from every parameter of MCMV infection. These included clinical signs, virus replication, histopathology and mortality.

The protective immunity was active against the virulent MCMV which was not neutralized by the rabbit anti‐attenuated MCMV serum.
Mechanism of Glucan‐Induced Agglutination in Streptococcus mutans*
Authors: Suginaka Hidekazu , Ebisu Shigeyuki , Kotani Shozo
Published: November 2013

The binding of radioactive glucan to Streptococcus mutatis cells, which are agglutinated by dextrans, was examined. The glucan was synthesized from sucrose by extracellular glucosyltransferases from S. mutans FA‐1 and was highly branched at C‐3 and C‐6 of D‐glucose residues, containing 17% of α(1→3)inter‐chain residues. Binding of glucan to whole cells of S. mutans OMZ‐176, which were agglutinated by addition of glucan or Dextran T2000, was irreversible and followed saturation type kinetics; saturation was achieved at approximately 110 ng of glucan per ml. About 14 ng of glucan were bound per mg of the cells at the saturated concentration. The heated cells of this organism, however, had a relatively low ability of glucan‐binding, compared with the freshly prepared and lyophilized cells. Binding to the heated cells was entirely of a non‐saturation type. Binding of Dextran T2000 or T10 was determined by competition between the labeled glucan and unlabeled Dextrans for the binding site(s). Both Dextrans and glucan from S. mutans FA‐1 were bound to the same site(s). Other organisms, which did not undergo glucan‐ and Dextran‐induced agglutination, had a relatively lower ability of glucan‐binding than S. mutans, which was agglutinated.
Effect of Double Infection of Cowpox Virus‐Infected Cells with Paramyxovirus (Sendai Virus) on Formation of Cowpox Virus‐Specific Cell Surface Antigen
Authors: Tanaka Junji , Ogura Hisashi , Fukuda Shizuo , Hatano Motoichi
Published: November 2013

The formation of cowpox virus‐specific cell surface antigen (CPV S‐ag) was significantly enhanced by double infection with HVJ (Sendai virus). Simultaneous double infection, superinfection with HVJ and superinfection with CPV of cells persistently infected with HVJ similarly enhanced the formation of CPV S‐ag, while pre‐infection with HVJ was ineffective. To be effective, cells must be infected at a m.o.i. of ≧ 1.0 and HVJ gene functions had to be expressed.

The HVJ‐infected cell extracts had an ability to accelerate uncoating (or degradation) of CPV, causing an early increase and a subsequent decrease in the infectivity of CPV. This activity reached a maximum 4–6 hr after HVJ infection, the increase paralleling enhancement of the total activity of several cellular enzymes. Addition of puromycin abolished the increase of these activities and the formation of CPV S‐ag.

Thus, the double infection with HVJ of CPV‐infected cells induces an enhancement of CPV S‐ag formation presumably as a consequence of activation of cellular enzymes which in turn accelerates uncoating of CPV.
Difference in Antibody Production to Heterologius Erythrocytes in Conventional, Specific‐Pathogen‐Free (SPF), Germfree and Antigen‐Free Mice
Authors: Taniguchi Tom , Suzuki Tatsuo , Miake Shunji , Nomoto Kikuo , Hashimoto Kazuo , Goda Akira , Takeya Kenji
Published: November 2013

The expression of antibody‐producing capacities against hamster erythrocytes (HRBC), known to be weakly immunogenic in mice, was compared among conventional, SPF, germfree and antigen‐free mice.

ICR strain germfree and antigen‐free mice showed antibody production to HRBC comparable to that in conventional or specific‐pathogen‐free (SPF) mice.

In the NC strain, some of the conventional mice produced low titers of antibody after a single injection of HRBC, but none of the germfree mice showed such a transient antibody production. In the ICR‐KIG strain, which was selected from the colony‐bred ICR strain, antibodies with high titers were produced after a single injection of HRBC under both conventional and germfree conditions. The onset of conversion from 2‐mercaptoethanol (2‐ME) sensitive to 2‐ME resistant antibody after a single injection of HRBC was not delayed in the germfree mice when compared with the conventional or SPF mice.

Antibody production to sheep erythrocytes (SRBC), known to be highly immunogenic in mice, was not influenced by exogeneous stimulation from microorganisms or diet. No differences in antibody production to SRBC were detected irrespective of the maintenance conditions of the mice.

Stimulation with microorganisms or diet may not be required as essential elements for the maturation of antibody‐producing capacities, but such a stimulation appears to modify the antibody response. The modifying effect was more prominent in the antibody response against weakly immunogenic antigens than against highly immunogenic ones.
Some Physicochemical Characteristics of an Immune Lymphocyte Product Which Inhibits the Multiplication of Toxoplasma within Mouse Macrophages
Authors: Shirahata Toshikazu , Shimizu Kiheiji
Published: November 2013

In response to antigenic stimulation, the splenic lymphocytes from Toxoplasma‐infected mice produce a factor which is called by the authors the Toxoplasma growth inhibitory factor (Toxo‐GIF) and which inhibits the multiplication of Toxoplasma within nonimmune macrophages in vitro. In this study, partial characterization of murine Toxo‐GIF was examined using Sephadex G‐100 gel‐filtration, isoelectric focusing, zonal electrophoresis and heat and enzymatic treatment.

Peak activity of Toxo‐GIF was found in a Sephadex G‐100 fraction with a similar molecular size to that of the ovalbumin. The molecular weight of Toxo‐GIF was calculated to be between 38,000 and 55,000. Toxo‐GIF was stable to heating at 56 C for 30 min but lost its activity at 80 C for 30 min or by exposure to pH values of 5 and 2. Exposure of Toxo‐GIF to water‐insoluble chymotrypsin or neuraminidase markedly decreased its ability to induce enhanced microbicidal activity of cultured macrophages, suggesting that Toxo‐GIF was a glycoprotein. Furthermore, Toxo‐GIF migrated in a region cathodal to mouse albumin on agar zone electrophoresis. Isoelectric focusing of active Sephadex fractions showed a well‐defined peak of Toxo‐GIF activity with an isoelectric point of pH 4.9 to 5.9.