bluuebunny/crossref_metadata_2025 huggingface dataset:
Sensitivity of Various H Particles of Poliovirus to Alkaline Treatment
Authors: Aikawa Shogo
Published: November 2013

Four kinds of H antigenic particles (H particles) of poliovirus were prepared, (1) by mild alkaline treatment of virions (Alkaline‐H), (2) by separation from the virus infected culture (Natural‐H), (3) by ultraviolet irradiation of virions (UV‐H), (4) by heating of virions (Heat‐H). They were examined for sensitivity to an alkaline degradation (pH 10.5 at 40 C). Heat‐H was most stable and no degradation of capsid was observed after the treatment. UV‐H was most unstable, since it was completely disrupted into small components after the treatment. Alkaline‐H and Natural‐H were relatively stable but a minor amount of small component(s) was liberated therefrom by the treatment. These results suggest that the H particles induced by the four procedures are different from each other in the stability of capsid structure. Furthermore, it is suggested that the antigenicity(s) of the small‐size component(s) is different from either N or H antigenicity being ascribable to VP2 polymer.
Tumor‐Specific Transplantation Antigen Activity of Freshly Adenovirus‐Infected Cells
Authors: Nakajima Setsuko , Hamada Chuya , Uetake Hisao
Published: November 2013

Newborn hamsters were inoculated with human adenovirus type 12 (Ad12) within 24 hr of birth for tumor induction, and 15 days later, intercurrently immunized with Ad12‐infected cells (KB; HeLa; FL; HEK; MoE; HaE). Tumor development was then observed for 75 days thereafter. Tumor formation was prevented at a statistically significant level by immunization with any of the above‐mentioned infected cells. The immunization was effective even with abortively infected cells (HaE; MoE) or with cells infected in the presence of 5‐fluorodeoxyuridine. The induced immunity was Ad12‐specific, since neither cells infected with Ad2, Ad7 or Ad18 nor CV‐1 cells infected with SV40 were able to prevent tumor formation. The most plausible explanation to these findings could be that Ad12‐specific tumor‐specific transplantation antigen is induced on the surface of freshly virus‐infected cells and it is responsible for induction of specific cellular immunity. This gives an experimental support to our hypothesis on the mechanism of induction of cellular immunity against virus infections and to the hypothesis proposed by Habel and by Sjögren to explain the immunoresistance against tumor cells induced following viral immunization.
Chemical Studies on the Cell Walls of Leptospira biflexa Strain Urawa and Treponema pallidum Strain Reiter
Authors: Azuma Ichiro , Taniyama Tadayoshi , Yamamura Yuichi , Yanagihara Yasutake , Hattori Yukio , Yasuda Seiji , Mifuchi Ichiji
Published: November 2013

The preparation and chemical poperties of the cell walls of Leptospira biflexa Urawa and Treponema pallidum Reiter are described. Both cell walls are composed mainly of polysaccharides and peptidoglycans. The data of chemical analysis indicate that the cell wall of L. biflexa Urawa contains rhamnose, arabinose, xylose, mannose, galactose, glucose and unidentified sugars as neutral sugars, and alanine, glutamic acid, α,ε‐diaminopimelic acid, glucosamine and muramic acid as major amino acids and amino sugars. As major chemical constituents of the cell wall of T. pallidum Reiter, rhamnose, arabinose, xylose, mannose, galactose, glucose, alanine, glutamic acid, ornithine, glycine, glucosamine and muramic acid have been detected. The chemical properties of protein and polysaccharide fractions prepared from the cells of T. pallidum Reiter were also partially examined.
The Metabolic and Phagocytic Activities of Leukocytes from Patients Receiving Corticosteroid and Radiation Therapy, and Patients with Bacterial Infections
Authors: Okuda Kenji , Tadokoro Ichiro , Noguchi Yoshikuni
Published: November 2013

Peripheral blood leukocytes from patients given corticosteroid or radiation therapy, as well as patients with bacterial or viral infections, were studied with regard to the selected enzyme activities of the hexose monophosphate shunt (HMS). Glucose‐6‐phosphate dehydrogenase (G‐6‐PD), 6‐phosphogluconate dehydrogenase (6‐PGD) and reduced nicotinamide‐adenine dinucleotide phosphate (NADPH) oxidase were assayed spectrophotometrically on mixed leukocyte suspensions in isotonic glycerol. Enzyme activities of G‐6‐PD and NADPH oxidase in patients receiving corticosteroid or radiation therapy were significantly lower than the enzyme activity of 6‐PGD. In patients with bacterial infections, activities of the three enzymes increased but in patients with viral infections, only the activities of NADPH oxidase and G‐6‐PD were slightly decreased. Nitroblue tetrazolium (NBT) dye‐reducing activities of neutrophils from patients receiving corticosteroid or radiation therapy were attenuated which coincides with the reduced activities of HMS enzymes. From these results, it is likely that the reduced activities of intraleukocytic HMS enzymes of patients receiving corticosteroid or radiation therapy are correlated with intracellular bactericidal activities which might result from the attenuated level of hydrogen peroxide production.
Relation between Enhanced Antibody Responses Elicited by Adjuvant Injection and Those Elicited by Secondary Antigenic Stimulation
Authors: Tanabe Masao J. , Saito Kazuhisa
Published: November 2013

An attempt was made to determine if there is any common mechanism in the enhanced antibody response caused either by injection of adjuvant, such as bacterial endotoxin (LPS) and complexed polynucleotides, or by secondary antigenic stimulation. LPS inoculated in mice 4 days before injection of sheep red blood cells (SRBC) and polyA:U invalidated the adjuvant effect of polyA:U injected together with SRBC, and the hemolysin plaque‐forming cell (PFC) response of such mice was similar to that of the mice which received SRBC alone. When mice primed with SRBC 24 days in advance were injected with LPS and 4 days later re‐stimulated with SRBC, their PFC response to the secondary stimulation was suppressed to less than one tenth of the normal secondary PFC response. The suppressive effect of LPS on the secondary antibody response was abolished if the serum collected from mice injected with LPS was given to the primed and LPS‐injected mice at the time of the secondary antigenic stimulation. From these results we discussed the possibility that some common mediator might play a role in the enhanced antibody response elicited by either adjuvant injection or secondary injection of antigen.
Inhibition of Macrophage Migration by Virus‐Induced Interferon Preparations1
Authors: Imanishi Jiro
Published: November 2013

It was found that a preparation of mouse L cell interferon induced by Newcastle disease virus (NDV) possessed not only interferon activity but also inhibitory activity upon migration of guinea pig peritoneal macrophages (MIF activity). These activities were also observed in a preparation of human leukocyte interferon induced by NDV. The interferon and MIF activities shared common characteristics in the dose response, time course of in vitro production, thermal stability, sensitivity to trypsin and periodate, and elution pattern in CM‐Sephadex column chromatography. However, gel filtration pattern with Sephadex G‐100 showed two separate peaks. Fractions collected from the first peak, corresponding to a molecular weight of about 45 000, had only the MIF activity, while those collected from the second peak, corresponding to a molecular weight of about 30 000, had both the interferon and MIF activities. A preparation of mouse brain interferon induced by Japanese encephalitis virus had a much weaker MIF activity than the L cell interferon, although these preparations were equal in interferon activity (5000 units/ml).
Drug Resistance in Staphylococcus aureus
Authors: Ono Hideo , Inoue Matsuhisa , Mao James C.‐H. , Mitsuhashi Susumu
Published: November 2013

Antibacterial and inducer activities concerning inducible macrolide resistance in Staphylococcus aureus were investigated using 32 erythromycin, oleandomycin and other macrolide antibiotic derivatives and analogues. The macrolides were classified into five groups from very high to none according to their inducer activity.
Mutants of Escherichia coli Which Block Head Formation of Lambda
Authors: Hidaka Saburo , Matsubara Kenichi
Published: November 2013

Five mutants of Escherichia coli K12 (lam 24, lam 25, lam 26, lam 27 and lam 646) that block head formation of λ are described. In vitro complementation tests and electron microscopy demonstrated that in these bacteria phage tails were produced normally, whereas head formation was abnormal, aberrant head‐related structures being produced. In lysates prepared from lam 24, lam 25 and lam 26, monsters and empty heads without tail were the predominant structures, whereas in lysates from lam 27 and lam 646, petit λ and empty heads were the most common structures. The five lam mutations were located in two regions on the bacterial chromosome; lam 24, lam 25 and lam 26 were near the dnaB gene and lam 27 and lam 646 near the lac gene. It was suggested that the former three mutants are new isolates that belong to GroE mutants, whereas the latter two comprise a new group of mutants. Analyses of phage mutants (ov mutants) that overcome the interference by the lam 646 mutation revealed that this mutation blocks normal expression of the gene E of λ.
Effect of Bacterial Lipopolysaccharides on Anti‐Trinitrophenyl Antibody‐Producing Cells
Authors: Saito Tatsuo , Nakano Masayasu
Published: November 2013

The effect of lipopolysaccharide (LPS) on anti‐trinitrophenyl (TNP) direct plaque‐forming cells (PFC) in the spleen of mice and the affinity of antibodies produced by these PFC were examined. Simultaneous injection of bacterial LPS and TNP‐coupled sheep red blood cells(SRBC) induced an obvious increase in anti‐TNP PFC numbers and heightened the antibody affinity at cellular levels. The higher the doses of LPS, the greater the effects. Concomitant injection of LPS in TNP‐coupled homologous mouse red blood cells (MRBC) also elicited good anti‐TNP PFC response and slightly heightened the affinity. Priming with LPS and SRBC together 7 days prior to immunization did not enhance the anti‐TNP PFC response and it was difficult to alter the affinity. Preinjection with small amounts of TNP‐MRBC or ‐rabbit red blood cells and LPS simultaneously did not induce any significant increase in anti‐TNP PFC secondary response after reimmunization with TNP‐SRBC, but obviously heightened the antibody affinity. Injection of LPS simultaneously with the secondary immunization was effective for both the anti‐TNP PFC response and the alteration of antibody affinity. These results suggest that LPS affects the control mechanisms of anti‐TNP antibody affinity via the non‐thymus‐derived helper cell function, and the adjuvant action and alteration of antibody affinity induced by LPS are regulated by different mechanisms.
Multinucleated Giant Cell Formation in BHK‐21‐528 Cell Monolayers Infected with Japanese Encephalitis Viruses1
Authors: Ueba Noboru , Kimura Tomoaki , Kimoto Tatsuo
Published: November 2013

Formation of prominent multinucleated giant cells (MGC) was observed in monolayers of a clonal line of BHK‐21 cells (BHK‐21–528) when infected with Japanese encephalitis virus (JEV). MGC were first observed 3 to 4 days after infection and cytopathic changes proceeded thereafter. Formation of MGC is a typical cytopathic change in this clonal cell line. Virus titer in 50% tissue culture infective dose (TCID50) equaled that in 50% MGC‐forming dose. Virus titer in TCID50 was approximate to plaque‐forming units (PFU) in the same host cells. An ability of JEV to form MGC was maintained at least for six serial passages in BHK‐21–528. It was inactivated by heating at 56 C for 3 min. All JEV strains, except an attenuated live vaccine strain, induced formation of MGC in BHK‐21–528 cells. Red blood cells of several animal species were not adsorbed to MGC induced by JEV. The MGC‐forming ability of JEV was specifically neutralized by anti‐JEV serum. By fluorescence antibody technique, the MGC were specifically stained by anti‐JEV antibody conjugated with fluorescein isothiocyanate. Immunization of animals with lysates of the MGC resulted in production of antibodies against JEV, but no antibody against other viruses which have been reported to induce MGC formation. From these evidences, it was concluded that JEV induced formation of MGC in BHK‐21–528.
Characterization of Background Anti‐Trinitrophenyl Plaque‐Forming Cells Observed in Several Strains of Mice
Authors: Uchiyama Takehiko , Yamaura Noboru , Maeda Ryozo
Published: November 2013

Normal mice have a large number of background anti‐trinitrophenyl (TNP) antibody‐forming cells (AFC) in their spleens (about 40–50 anti‐TNP PFC/106 cells). We investigated this among several mouse strains, i.e., C57BL/6, C3H/He, Balb/c, ddd, and ICR mice, and found that all strains had a similar number of anti‐TNP PFC (plaque‐forming cells). Developmental aspects of background anti‐TNP PFC in the ontogenic process were also investigated. The number of anti‐TNP PFC increased logari thmically during the first few days of age, reached a peak on the 13th day and attained a constant value within 30 days. Neonatal thymectomy did not decrease the number of background anti‐TNP PFC but such treatment decreased the anti‐TNP PFC response to TNP‐HRBC (horse red blood cells) immunization. Germ‐free ICR mice had a number of background anti‐TNP PFC similar to that of conventional ICR mice. Avidity of background anti‐TNP PFC was compared among mice of several ages and it was shown that there were no differences among them. These results suggest that the occurrence of these background anti‐TNP PFC is not elicited by the immune response but by the natural maturation of precursors of AFC without antigenic stimulation.
Strain Differences in the Immunogenicity of Aggregated Human IgG and the Adjuvant Action of Lipopolysaccharide on the Low‐Responder Strain of Mice
Authors: Fujiwara Michio , Fujiwara Shigeko , Yoshizaki Chiho , Awaya Akira
Published: November 2013

Strain differences in the antibody response to human IgG (HGG) were observed when aggregated HGG was injected intravenously. Lipopolysaccharide (LPS) administered subsequently markedly enhanced the antibody response to HGG in low responder C57BL/6 mice as compared with that in high responder DDD, C3H/He or (C57BL/6 × DDD)F1 mice. Aggregate‐free preparation of HGG at a dose of 0.5 mg induced immunological tolerance in all strains of mice tested. LPS injected subsequently converted tolerogenic, aggregate‐free HGG into immunogen in DDD mice but not in C57BL/6 mice. To determine the correlation between adjuvanticity and mitogenicity of LPS, spleen cells from normal mice were cultured in the presence of LPS and 3H‐thymidine uptake was measured. Spleen cells of DDD mice incorporated three times as much 3H‐thymidine as those of C57BL/6 mice. There seems no strong correlation between both activities of LPS. The data obtained are discussed in terms of strain differences in the macrophage function for processing the antigen.
Effect of Capsular Polysaccharide of Klebsiella pneumoniae on Host Resistance to Bacterial Infections
Authors: Kato Nobuo , Kato Osamu , Nakashima Izumi
Published: November 2013

When Klebsiella pneumoniae capsular polysaccharide (CPS‐K) from type 1, Kasuya strain, was injected intraperitoneally (i.p.) immediately before i.p. bacterial challenge, the survival time of mice infected with Salmonella enteritidis NUB 1 (virulent strain) was shortened and the mortality rate for mice infected with S. enteritidis NUB 31 (avirulent strain) was enhanced. The promotion of infection with S. enteritidis NUB 1 by CPS‐K depended upon its dose, the effect of CPS‐K being demonstrable up to as little as 0.2 μg per mouse. In the case of S. enteritidis NUB 31, the effect of CPS‐K was detectable only when more than 20 μg per mouse was injected. As a result of enumeration of bacterial populations in the peritoneal washing, blood, liver and spleen, it was revealed that CPS‐K promoted in vivo growth of S. enteritidis NUB 1 and NUB 31. In addition, CPS‐K enhanced the mortality rate in mice infected with Streptococcus pyogenes or Streptococcus pneumoniae. The peak CPS‐K effect on infection with S. enteritidis NUB 1 was seen when given immediately before bacterial challenge. The active substance responsible for the infection‐promoting effect of CPS‐K was neutral CPS‐K, which is distinct from the O antigen and from acidic CPS‐K (the type‐specific capsular antigen). Preparations of neutral CPS‐K isolated from the other three strains of K. pneumoniae exhibited a marked infection‐promoting effect comparable with that of preparations from the Kasuya strain. Neutral CPS‐K, with identical antigenicity to that from the Kasuya strain, has already been found to exert a strong adjuvant effect on antibody responses to various antigens in mice. No parallelism exists between infection‐promoting activity and adjuvant activity of neutral CPS‐K.
An Improved Procedure of Specific Polysome Precipitation with Antibody
Authors: Ono Masao , Kawakami Masaya
Published: November 2013

The specific immunoprecipitation of polysomes prepared from a mouse myeloma, 31C, synthesizing an IgG1 immunoglobulin has been investigated. A reported method in which polysomes were coprecipitated by sequential addition of antibody to 31C myeloma protein, antigen (i.e., the 31C protein) and again the antibody, was used. Salt concentration greatly affected the immunoprecipitation of polysomes. In the presence of 100 mm KCl or NaCl, 10–20% of myeloma polysomes and only 1% of mouse liver polysomes were precipitated with the antibody to myeloma protein. On the other hand, 90% of the both polysomes were precipitated by the same antibody at a salt concentration of 10 mm. Triton X‐100 and sucrose had little effect on preventing nonspecific binding of immunoglobulin to ribosomes. Experiments were carried out to obtain an optimal ratio of the amount of polysome to that of antibody and antigen to be added for the coprecipitation of polysomes. To date we have tried 25 μg of the first antibody, 14 μg of antigen added second to the polysomes and 38 μg of the antibody added finally and these were found to precipitate most efficiently one A260 unit of 31C polysomes.
Mouse Virulent Strain of Staphylococcus epidermidis
Authors: Yoshida Kosaku , Ichiman Yoshitoshi , Ohtomo Toshichika
Published: November 2013

Using 109 or 107 colony‐forming units of a strain of Staphylococcus epidermidis (strain 1142) in saline or 5% mucin, respectively, 90 to 100% of mice died within 24 to 48 hr after intraperitoneal challenge infection. These organisms gradually multiplied in the peritoneal cavity when injected intraperitoneally into mice, while the mouse avirulent strain (strain 1124) rapidly decreased and no organisms were found there 20 hr after injection. This strain was capable of inducing resistance against challenge with homologous strains. The resistance appeared as early as the first week and disappeared the 4th week after the immunization. However, no resistance was induced with strain 1124 against challenge with strain 1142. Also, hyperimmune rabbit serum prepared with strain 1142 passively protected against challenge with homologous strain in mice. The protective antibody was absorbed out with homologous organisms but not with strain 1124. Subsequently, a surface substance was obtained from strains 1142 or 1124 by the method of Morse. The 1142 surface substance was capable of inducing a resistance against challenge with the homologous strain but not with the 1124 surface substance. Also, this substance absorbed the protective antibody in hyperimmune rabbit serum prepared with the homologous strain but not with the 1124 surface substance nor with the Smith surface antigen extracted from the Smith strain of Staphylococcus aureus. Conversely, the protective antibody in rabbit anti‐Smith strain serum against challenge with the homologous strain was absorbed with the Smith surface antigen but not with the 1142 surface substance. In the agar diffusion test, the 1142 surface substance and the Smith surface antigen produced single precipitin lines only against homologous antisera. Biochemical analysis of the 1142 surface substance showed that the substance contained neither nucleic acids nor proteins but is composed of hexosamine, glycerol, phosphorus, alanine, glycine and phenylalanine.
Biological Activities of Endotoxins from Yersinia enterocolitica
Authors: Kanamori Masato
Published: November 2013

The chemical properties and the general biological activities of lipopolysaccharide (LPS) and Boivin‐type endotoxin obtained respectively by phenol‐water and trichloroacetic acid extraction from Yersinia enterocolitica serotypes 03 and 09 were studied. The yield of LPS from the O9 strain was about 10% of the O3 strain possibly because of the lower solubility of O9‐LPS in aqueous phase. However, the chemical composition of O9‐LPS was similar to that of O3‐LPS in the proportions of reducing sugar, glucosamine, heptose, KDO, and lipid A. In pyrogenicity and local Shwartzman reactivity in rabbits and lethality for mice, there was also no difference between O3 and O9‐LPS. The anthrone‐positive carbohydrate and lipid A contents of Boivin‐type endotoxin from O3 were higher than those of the endotoxin from O9. The biological activities of Boivin‐type endotoxin from O3 were also remarkably higher than those of the endotoxin from O9. It seems that endotoxin of Y. enterocolitica serotype O3 may play an important role in infection by this organism.
Quantitative Chemical Analyses and Antigenic Properties of Peptidoglycans from Clostridium botulinum and Other Clostridia
Authors: Takumi Kenji , Kawata Tomio
Published: November 2013

The cell wall peptidoglycans were isolated from Clostridium botulinum and some other species of the genus Clostridium by hot formamide extraction and their quantitative chemical composition and antigenic properties were determined. The peptidoglycan of C. botulinum type E was found to be a diaminopimelic acid (DAP)‐containing type composed of glucosamine, muramic acid, glutamic acid, alanine and DAP in the molar ratio of 0.76:0.78:1.00:1.88:0.81. All other types of C. botulinum and Clostridium sporogenes also belonged to the same peptidoglycan type. The peptidoglycans of Clostridium bifermentans and Clostridium histolyticum contained DAP but they differed from those of C. botulinum in the molar ratio of alanine to glutamic acid. The peptidoglycan of Clostridium perfringens was composed of glutamic acid, alanine, DAP and glycine in the molar ratio of 1.00:1.64:0.94:0.90. On the other hand, the peptidoglycan of Clostridium septicum was found to contain lysine instead of DAP and the molar ratio was 1.00:1.41:0.96 for glutamic acid, alanine and lysine. In spite of the difference in amino acid composition of peptidoglycans among the Clostridia, the quantitative precipitin test demonstrated that antiserum against C. botulinum type E peptidoglycan cross‐reacted with the peptidoglycans from other Clostridia as well as various types of C. botulinum.
Effects of BCG (Bacillus Calmette–Guérin) Vaccines on Immune Responses in Mice
Authors: Kitamura Yukikazu , Nomoto Kikuo , Torisu Motomichi , Takeya Kenji
Published: November 2013

The effects of killed and living BCG on antibody production against hamster erythrocytes (HRBC) and the 2, 4, 6‐trinitrophenyl (TNP) group were studied in SL mice. Killed and living BCG, each in doses of 0.008 mg, 0.08 mg, 0.8 mg and 8 mg per mouse, were intravenously inoculated 7 days prior to primary immunization with HRBC. Secondary immunization was carried out 28 days later with TNP‐HRBC. Anti‐HRBC and anti‐TNP antibodies were estimated by a hemagglutination test. The results showed that pretreatment with killed or living BCG enhanced the antibody production against both HRBC and TNP. Comparing the effects of these two BCG preparations, it was noted that killed BCG augmented the anti‐HRBC antibody production more effectively than living BCG. In regard to the anti‐TNP antibody production, living BCG exhibited a greater augmenting effect than killed BCG. This difference in the modes of action of killed and living BCG was remarkable when two groups given 8 mg of killed and living BCG were compared. In addition, it was shown that living BCG at a dose as high as 8 mg was able to augment the anti‐TNP antibody production, even in the absence of preceding immunization with HRBC.
In Vitro Studies on the Mechanism of Acquired Resistance to Tuberculous Infection
Authors: Muraoka Shizuko , Takeya Kenji , Nomoto Kikuo
Published: November 2013

Immune lymph node cells were obtained from mice immunized with bovine gamma globulin (BGG) in complete Freund's adjuvant or allogeneic MH134 tumor cells. They showed the capacity of conferring bactericidal activity on macrophages infected with Mycobacterium tuberculosis, H37Rv, when they were incubated on macrophage monolayers together with the corresponding antigen, i.e., BGG or solubilized cellular antigen of the tumor cells. However, such capacity was lower than that of tubercle bacilli‐immune lymph node cells. Culture supernatants were harvested after incubation of tubercle bacilli‐immune, BGG‐immune or allogeneic tumor‐immune lymph node cells with the corresponding antigen for 24 hr. Macrophages were altered so as to suppress intracellular bacillary growth when macrophage monolayers were exposed to the supernatants for more than 2 days. When normal lymph node cells were incubated on normal macrophage monolayers together with a mitogen such as PHA or concanavalin A, growth of tubercle bacilli within the macrophages was slightly but difinitely suppressed. The mechanism of elicitation of cellular immunity to the infection with tubercle bacilli is discussed on the basis of results presented in this and the preceding paper.
Grouping Antigens of Four Lactobacillus Species and Their Characteristics
Authors: Shimohashi Hirotaka , Kodaira Shinji , Suegara Nobuo
Published: November 2013

Antigenic analyses of Lactobacillus bulgaricus, Lactobacillus lactis, Lactobacillus brevis and Lactobacillus buchneri were carried out by double immunodiffusion in agar. Antigens were extracted from whole cells and cell wall preparations with cold trichloroacetic acid. Most strains of the four species possessed antigen 9 in their cell walls. Another antigen, antigen 10, was found in the cell walls of all the strains of L. brevis and L. buchneri, and in some strains of L. lactis, but not in L. bulgaricus. Fractionation of the antigens was attempted using the cell wall extracts of L. lactis L‐10 with only antigen 9 and of L. brevis X‐1 with both antigens 9 and 10. The partially purified fractions of antigen 9 and of the complex of antigens 9 and 10 were obtained by zone electrophoresis. However, antigen 10 from the complex could not be separated by the same method or gel filtration on Sephadex G‐100 since the two antigens 9 and 10 of the complex always behaved together. The fraction of antigen 9 consisted almost entirely of glycerol and glucose as sugar components, the molar ratio being 2:1. The complex of antigens 9 and 10 also consisted of the same sugars, and the molar ratio of glycerol: glucose was 4:1. Inhibition tests indicated that the immunodominant component of antigen 9 was α‐methylglucoside (glucose), and most probably the determinant is a glucosylated glycerol teichoic acid. It was considered that the determinant of antigen 10 is a glycerol teichoic acid although glucosamine and galactosamine inhibited effectively the reaction between antigen 10 and its antibody.
Ineffectiveness of Lipopolysaccharide for Preventing the Tolerance Induction in Bone Marrow‐Derived Lymphoid Cells with Dinitrophenyl‐Poly‐l‐(Glutamic Acid, Lysine)
Authors: Uchiyama Takehiko , Yamaura Noboru
Published: November 2013

The effect of endotoxin or lipopolysaccharide (LPS) on tolerance induction in bone marrow‐derived lymphoid cells (B cells) was investigated. Dinitrophenylated amino acid copolymer‐l‐(glutamic acid, lysine) (DNP‐GL) acts as a potent tolerogen on normal and DNP‐primed B cells. LPS significantly enhanced the anti‐sheep red blood cell plaque‐forming cell (anti‐SRBC PFC) response that occurred after the immunization with a low dose of SRBC. LPS did not induce the primary anti‐DNP PFC response after the injection of DNP‐GL, nor did it prevent the tolerance induction in normal and DNP‐primed B cells that occurred after the administration of DNP‐GL.
Heterogeneity of Staphylococcal Enterotoxin A on Isoelectric Focusing and Disc Electrophoresis
Authors: Yamada Sumio , Igarashi Hideo , Terayama Takeshi
Published: November 2013

Heterogeneity of purified staphylococcal enterotoxin A, obtained from a culture supernatant of Staphylococcus aureus, strain 13N‐2909, was demonstrated by isoelectric focusing. The toxin was composed of three immunologically identical fractions with isoelectric points of 6.5, 7.0 and approximately 8.0. Heterogeneity of the toxin was also shown by disc electrophoresis. At pH 8.0 and 9.4 two major bands and a faint minor band were observed, while at pH 4.3 only one band was observed. The faster‐moving band for the anode in disc electrophoresis at pH 9.4 was found to correspond with the pI 6.5 component from isoelectric focusing, while the slower‐moving band corresponded with the pI 7.0 component. From the results of the electrophoretic migration tests of the toxin, the components corresponding to the two major bands found in disc electrophoresis at pH 9.4 were considered to be charge isomers.
Distribution of Clostridium botulinum Type C in Ishikawa Prefecture, and Applicability of Agglutination to Identification of Nontoxigenic Isolates of C. botulinum Type C
Authors: Serikawa Toshihiko , Nakamura Shinichi , Nishida Shoki
Published: November 2013

Since deaths of waterfowls have frequently been observed in Lake Kahoku near Kanazawa city, Japan, we attempted an ecological study on Clostridium botulinum type C in four other lakes as well as Lake Kahoku. One hundred and twenty‐nine (56%) of 230 soil samples collected gave rise to lethal toxicity in mice with the characteristic “wasp‐waist” symptom. All of the 51 samples arbitrarily selected were neutralized by C. botulinum type C antitoxic serum. A further seasonal study throughout the year at a given shore area of Lake Kahoku disclosed that nearly all samples gave rise to toxicity due to C. botulinum type C during the autumn season when the most waterfowls congregate. Toxigenic strains of C. botulinum type C were isolated together with nontoxigenic strains that were culturally and biochemically similar to the toxigenic strains. Both the toxigenic and nontoxigenic strains were equally agglutinable by an antiserum prepared against one of the nontoxigenic strains. Further extensive studies on the specificity of the agglutination method for identification were performed with 112 strains of 46 clostridial species. None of the strains used except some strains of C. novyi type A and a strain of C. botulinum type D was agglutinable. Based on the findings for cultural, biochemical, and agglutinable properties, the nontoxigenic strains were identified as C. botulinum type C. Also, C. novyi type A isolates showing colonies covered with a small pearly layer zone but surrounded by an aberrantly wide lecithinase zone are discussed.
Effect of Various Reagents on the Susceptibility of Target Cells to the Cytotoxicity of Guinea Pig Lymphotoxin (GLT)1
Authors: Sawada Jun‐Ichi , Kishie Takao , Osawa Toshiaki
Published: November 2013

The cytotoxic activity of guinea pig lymphotoxin (GLT) was increased when the target cells were treated with the following metabolic inhibitors: puromycin, actinomycin D, NaN3, NaF, and 2,4‐dinitrophenol, whereas calf serum, N‐acetyl‐D‐galactosamine, colchicine, and vinblastin inhibited GLT‐cytotoxicity. Puromycin, an inhibitor of protein synthesis, exhibited the most potent enhancing effect on GLT‐cytotoxicity. It seems likely that there are certain cellular metabolic processes depending on cellular protein synthesis which antagonize GLT‐cytotoxicity. Taking advantage of this effect of puromycin, a time‐saving assay method for GLT‐cytotoxicity was developed.
An Analysis of Factors Influencing the Isolation Rate of Herpes Simplex Virus
Authors: Tada Aiko , Serine Noriyoshi , Toba Masanori , Yoshino Kamesaburo
Published: November 2013

Attempts were made to improve the rate of isolation of herpes simplex virus (HSV) from clinical specimens by minimizing loss of virus infectivity during transportation and employing the most sensitive cells for isolation. Basical analyses using standard strains of type 1 and type 2 HSV indicated that virus titer decrease was marked even at low temperatures in environments free of proteinous stabilizer such as normal serum or tissue extract, negating the generally held concept that HSV is stable in distilled water. YLE (Earle‐lactalbumin hydrolysate‐yeast extract) medium containing 20% inactivated calf serum was determined to be a transport medium of choice, because degradation of suspended virus during storage and freeze‐thawing was negligible and loss of virus during Millipore filtration was minimal. Special coating of the membrane could also be obviated by the use of this solution. In a cell susceptibility test using clinical specimens, secondary rabbit kidney (SRK) cells were the most sensitive, showing a quick development of cytopathic effect. Vero and RK‐13 cells were the second best, whereas monkey kidney, HeLa and L cells were far less sensitive. A total of 136 specimens from suspected cases, sent by dermatologists, were tested using SRK cells, and 99 strains of type 1 and 15 strains of type 2 HSV were isolated. Excluding one case from which vaccinia virus was isolated, the isolation rate of HSV was 84.4%.
Characterization of Simian Virus 40 Transformed African Green Monkey Cells (CV‐1)
Authors: Hirai Kanji
Published: November 2013

Several clones of SV40 transformed CV‐1 cells have been characterized for the production of T‐ and V‐antigens and for the state of viral genome. The transformed CV‐1 cells failed to produce infectious virions as assayed after sonication or cocultivation and fusion with normal CV‐1 cells, and were resistant to superinfection by SV40. Some clones of the transformed cells contained V‐antigens. The population of V‐antigen positive cells varied from 0 to 100% depending on the passage number while the T‐antigen positive cells were always 100%. The virions isolated from the transformed cells were similar in morphology to complete SV40, but lighter in density than complete SV40. In one clone, a small amount of SV40 DNA was detectable in a free state while a large proportion of the DNA hybridizable with SV40 3H cRNA was linearly integrated into the cell DNA. The free SV40 DNA was noninfectious, closed circular DNA with a size smaller than infectious SV40 DNA component I. Since the cell extracts of the transformed cells contained an agent(s) which induced T‐ and V‐antigens in normal CV‐1 cells, it was suggested that the SV40 transformed CV‐1 cells contained free as well as integrated defective SV40 genomes responsible for the synthesis of T‐ and V‐antigens.
Antiviral Effects of Double‐Stranded RNA from Rice Dwarf Virus on Infection of Mice with Western Equine Encephalitis Virus
Authors: Takehara Manabu
Published: November 2013

A single administration (1 to 10 mg/kg) of rice dwarf virus RNA (RDV‐RNA) prior to virus challenge reduced the mortality of mice infected with western equine encephalitis (WEE) virus. The protective effect of RDV‐RNA was significantly higher than that of polyinosinic‐polycytidylic acid. However, when these RNAs were given after virus infection, the protective effect was negligible. The titer of circulating interferon in mice reached a peak about 5 hr after injection of these RNAs and remained at this level for about 24 hr. Viremia in mice infected with a lethal dose of WEE virus was markedly suppressed by the treatment of mice with these RNAs. A pathological examination of mice treated with a lethal dose of RDV‐RNA revealed marked changes including degeneration and karyorrhexis in the lymphoid tissues of the spleen.
Studies on the Pathogenicity ofYersinia enterocolitica
Authors: Une Tsutomu
Published: November 2013

Analysis of the pathogenicity ofYersinia enterocoliticawas performed by means of cell culture studies on the interaction of the organisms with HeLa cells and rabbit peritoneal macrophages based on observations of the pathogenic behavior of the organismsin vivo(11). The pathogenic strains, which successfully produced experimental enterocolitis in rabbits (11), had the ability to penetrate HeLa cells and to survive or multiply within the macrophages. The nonpathogenic strains, lacking the ability to produce pathological changes in rabbits (11), failed to penetrate HeLa cells, except for one strain, and also to survive totally or multiply within the macrophages. It was evident that the abilities of the organisms to penetrate epithelial linings which serve as the barrier of intestinal mucosa and to survive or multiply within the host cells appears to be closely related to the pathogenicity ofY. enterocolitica.
Restitution of Hemagglutinating Activity to Spikeless Particles of HVJ (Sendai Virus) by Glycoprotein Components of Newcastle Disease Virus
Authors: Iinuma Masao , Maeno Koichiro , Matsumoto Toshisada , Hoshino Munemitsu
Published: November 2013

Spikeless particles of HVJ (Sendai virus) lacking in hemagglutinating (HA) activity were obtained by enzymatic digestion of virions with trypsin followed by centrifugation through a sucrose gradient. When they were mixed with glycoprotein components of Newcastle disease virus (NDV) obtained by treatment of purified virions with deoxycholate (DOC), the mixture showed hemagglutination reaction, which was inhibited by anti‐NDV serum, but not by anti‐HVJ serum. Sedimentation profile of the HA active agents was then examined by centrifugation of the mixture of spikeless particles of HVJ (labeled with 3H‐uridine) and glycoproteins of NDV (labeled with 14C‐amino acid mixture). The results showed that the peak of HA activity had both of the radioactivities, and that the sedimentation rate of the HA was faster than that of spikeless HVJ but slower than that of intact HVJ. Electron micrographs of such HA active structures showed that they were morphologically closely similar to intact virion of HVJ, although they had neither hemolytic activity nor infectivity. The mixture of spikeless HVJ and glycoproteins of HVJ or NDV which were removed from virions by proteolytic enzymes, on the other hand, did not show any detectable hemagglutinating activity.
Gelation of Limulus Lysate by Synthetic Dextran Derivatives
Authors: Suzuki Masuko , Mikami Takeshi , Matsumoto Tatsuji , Suzuki Shigeo
Published: November 2013

The Limulus test has been considered specific for the presence of bacterial endotoxins. To synthesize a simple model of endotoxin, palmitoyldextran phosphate was prepared by modification of dextran by palmitoylation and phosphorylation. The present studies indicated that a variety of polysaccharide derivatives, such as palmitoyldextran phosphate, palmitoyldextran, and dextran phosphate, give a positive Limulus test and show pyrogenic activity, except for low molecular dextran derivatives. On the other hand, polysaccharides, such as dextran, starch (soluble), chitosan, xylan, and lentinan, were negative in these assays. The gelation reaction of Limulus lysate by modified dextran derivatives may depend on the molecular weight or modification of polysaccharides by palmitoylation and/or phosphorylation to a great extent.
Immunochemical Isolation of γ‐Globulin mRNA and Estimation of Immunoglobulin Gene Reiteration
Authors: Muto Masahiro
Published: November 2013

The polyribosomes synthesizing γ‐globulin have been isolated by the achievement of specific precipitation using bentonite‐treated anti‐IgG antibody. The RNA extracted from the immunochemically precipitated polysomes was tested for its ability to direct the synthesis of proteins in a cell‐free system. The specific γ‐globulin‐synthesizing activity (cpm of γ‐globulin synthesized/μg RNA) of this RNA was 10‐fold greater than that from total polysomes. γ‐globulin mRNA (messenger RNA) isolated by immunoprecipitation was more than 89% pure with respect to contamination by other species of mRNA. The products synthesized by the cell‐free system were also analyzed by sodium dodecyl sulphate(SDS)‐polyacrylamide gel electrophoresis.

This RNA has been hybridized with mouse myeloma DNA. The estimation of immunoglobulin gene reiteration was carried out using hybridization kinetics with consideration given to the DNA/RNA ratio since the estimation from the “half Cot value” is not accurate. The results suggest that in the mouse there are about 20 copies per subgroup of genes coding for the variable region of the H and L chains.
Effect of Dihydroxymethyl Furatrizine on Cell Division of Escherichia coli
Authors: Iida Kyoko , Koike Masaatsu
Published: November 2013

Antibacterial activities of 3‐di(hydroxymethyl) amino‐6‐[2‐(5‐nitro‐2‐furyl)vinyl]‐1, 2, 4‐triazine, (dihydroxymethyl furatrizine) were investigated using mutant strains of Escherichia coli lacking repair systems for DNA damage, i.e. polA, uvrA, uvrB, uvrC, recA, recB, recC and uvrArecA. All of the mutant strains were more sensitive to the drug than the parent strains, as was the case with the sensitivity to UV‐irradiation. These results indicate that the drug acts lethally on sensitive bacteria by damaging their DNA, and parts of the damaged DNA are repaired by excision and recombinational repair systems. Filamentous cell formation was induced in all strains except the uvrArecA strain by sublethal concentration of the drug, as well as by UV‐irradiation. It is possible that the occurrence of the short period of “unbalanced growth” induced by such DNA damaging agents leads to filament formation. In the cells of the double mutant, filament formation was induced by the drug but not by UV‐irradiation, and the majority of the filamentous cells formed were multinucleated. This suggests that, in this double mutant, the drug directly reacts with the septation machinery of the cell envelope, resulting in filament formation. This hypothesis is supported by the electron microscopic observations that septation is interrupted in the filamentous cells induced by the drug.
Comparative Studies on Cytopathic Effects Induced by Vesicular Stomatitis Virus in Two Cell Types
Authors: Nishiyama Yukihiro , Ito Yasuhiko , Shimokata Kaoru , Nagata Ikuya , Kurachi Noriko , Sugiura Yasuo
Published: November 2013

Infection of mouse L cells with vesicular stomatitis virus (VSV) leads to an extensive cell fusion, while porcine kidney stable (PS) cells infected with VSV show only cell rounding. Therefore, comparative morphological studies on the infection of the two cell lines were carried out using a transmission or scanning electron microscope and an immunofluorescence microscope. PS cells infected with VSV contrasted to L cells infected with the same virus in the following two points; (1) the principal site of VSV maturation was the intracytoplasmic vacuolar membrane in PS cells and the plasma membrane in L cells. However, it was found that viral glycoprotein was present on the cell surface of infected PS cells; (2) the morphological changes at the cell surface of infected PS cells occurred much earlier and were severer than those at the cell surface of infected L cells. From these observations, we discuss the possibility that the surface membrane of PS cells is too sensitive to the VSV‐induced cell damage to cause cell fusion.
Cellular Elements in the Resistance to Candida Infection in Mice
Authors: Miyake Tsunenori , Takeya Kenji , Nomoto Kikuo , Muraoka Shizuko
Published: November 2013

Live organisms (cfu) of Candida albicans per organ were counted 1 hr and 1 to 20 days after an intravenous inoculation into various groups of mice which had distinct levels of immunologic or non‐immunologic defense mechanisms. a) The number of cfu in the liver decreased progressively in normal mice, but those in the kidney maintained a constant level during the observation period. b) The number of cfu in the liver decreased progressively also in nude mice. In their kidneys, however, cfu increased progressively at a late stage of infection. c) In lethally irradiated AKR or nude mice in which phagocyte functions were severely depressed, the number of cfu increased progressively in both liver and kidney from the initial stage of infection. d) In immunized AKR mice, growth of C. albicans was suppressed at late stages of infection. Such protective immunity could be transferred partly with immune lymphoid cells but not with hyperimmune serum in the experimental system employed. In protection against Candida infection, non‐immune phagocytosis and T cell‐mediated immunity appear to be required at the early and late stages of infection, respectively.
Growth of Measles Virus in Various Human Lymphoid Cell Lines
Authors: Minagawa Tomonori , Sakuma Takashi
Published: November 2013

Neurovirulent TYCSA strain and attenuated Schwarz strain of measles virus and Halle strain of subacute sclerosing panencephalitis (SSPE) virus replicated in cultures of human lymphoid cell lines of the T‐cell type, MOLT‐3, MOLT‐4 and CCRF‐CEM. TYCSA and Halle strains grew rapidly, but Schwarz strain grew slowly in these cell lines. Furthermore, these three strains established persistent infection in CCRF‐CEM cells but not in the other cell lines. In these persistently infected cultures an almost entire population of cells were shown to be infected and infectious virus was produced constantly for over 100 days. Cells persistently infected with Schwarz strain contained nucleocapsid structures in both the nucleus and cytoplasm and produced low titered infectious virus, whereas nucleocapsid structures were observed only in the cytoplasm of cells persistently infected with either TYCSA or Halle strain and the titers of infectious virus produced from these cells were high.
Antibacterial and Inducer Activities for Tetracycline Resistance by Its Derivatives and Analogues
Authors: Inoue Matsuhisa , Kazawa Toshio , Mitsuhashi Susumu
Published: November 2013

Antibacterial and inducer activities of thirteen TC derivatives were investigated for tetracycline (TC) resistance in staphylococcus aureus. Four compounds of the TC derivatives were not able to induce the resistance to TC in Staphylococcus aureus MS3937 rms7 (TC)+ harboring an inducible TC resistance determinant located on a plasmid. The 12a‐hydroxy position seems to be essential for the inducer activity.
Resistance Mechanism of Chloramphenicol in Streptococcus haemolyticus, Streptococcus pneumoniae and Streptococcus faecalis
Authors: Miyamura Sadao , Ochiai Hiroshi , Nitahara Yoshiyuki , Nakagawa Yoji , Terao Michinori
Published: November 2013

The chloramphenicol resistance of Streptococcus haemolyticus, Streptococcus pneumoniae and Streptococcus faecalis isolated from clinical materials was proved to be due to an inactivating enzyme produced by these bacteria. The inactivated products of chloramphenicol were identified as 1‐acetoxy, 3‐acetoxy and 1,3‐diacetoxy derivatives by thin‐layer chromatography and infrared spectroscopy. The responsible enzyme was thus confirmed to be chloramphenicol acetyltransferase. The enzyme was inducible. It was partially purified by ammonium sulfate precipitation, DEAE‐cellulose chromatography and gel filtration on Sephadex G‐150. The enzymes obtained from S. haemolyticus, S. pneumoniae and S. faecalis have been compared with the conclusion that they are identical with respect to molecular weight (approximately 75,000–80,000), optimum pH and heat stability.
Detection of R Plasmids in Naturally Occurring Fish‐Pathogenic Bacteria, Edwardsiella tarda
Authors: Aoki Takashi , Arai Toshihiko , Egusa Syuzo
Published: November 2013

The conjugative R plasmids were detected from fish‐pathogenic Edwardsiella tarda isolated from cultured eels. These R plasmids were resistant to sulfonamide and tetracycline, or sulfonamide, streptomycin, chloramphenicol, tetracycline and kanamycin. There were no inhibitions in growth of phages W‐31, P1, T1, T3, T7, and λ. They were classified into incompatibility group A.