bluuebunny/crossref_metadata_2025 huggingface dataset:
Interferon and Cytotoxic Factor (Cytotoxin) Released in the Blood of Mice Infected with Mycobacterium bovis BĆG
Authors: Kato Nobuo , Nakashima Izumi , Ohta Michio , Naito Setsuko , Kojima Takashi
Published: November 2013

Interferon production stimulated by the active substance (neutral fraction) of the capsular polysaccharide of Klebsiella pneumoniae (neutral CPS‐K) in BCG‐infected mice was compared with that by bacterial lipopolysaccharide (LPS). Prior infection with BCG increased the responsiveness of mice to the lethal effect of neutral CPS‐K as well as to that of LPS. Associated with this, BCG‐infected mice showed a markedly enhanced ability to produce interferon after stimulation not only by LPS but also by neutral CPS‐K. In addition, a cytotoxic factor (cytotoxin) was found to be released in the serum of BCG‐infected mice after injection of these inducers. The kinetics of production of interferon and cytotoxin stimulated by neutral CPS‐K were very similar to those stimulated by LPS. The time pattern of cytotoxin production was not in parallel with that of interferon production. Interferon reached a peak 2 hr and cytotoxin 3 hr after injection with these inducers. Interferon and cytotoxin produced by neutral CPS‐K showed essentially the same stabilities to heating at 56 C and to treatment at pH 2 respectively as those produced by LPS. Interferon was inactivated by heating at 56 C more rapidly than cytotoxin. Cytotoxin was inactivated by treatment at pH 2 for 24 hr, whereas interferon activity was well preserved after this treatment. These results suggest that both activities are the result of different substances.
Further Studies on Generation of Macrophages in In Vitro Cultures of Mouse Spleen Cells and Its Inhibition by the Capsular Polysaccharide of Klebsiella pneumoniae
Authors: Yokochi Takashi , Nakashima Izumi , Kato Nobuo
Published: November 2013

Although the number of macrophages detected in cultures of mouse spleen cells at the start of the culture was very small, it markedly increased during further incubation. Macrophages were generated not only from the glass‐adherent cell fraction of spleen cells, but also from the nonadherent cell fraction obtained after removal of adherent cells either by incubating in glass petri dishes or by passing through a glass bead column. The generation of macrophages from the nonadherent cell fraction occurred even when it was separated as late as 48 hr after the start of the culture. The phagocytic activity of macrophages newly generated from the nonadherent cell fraction was relatively weak, but it was activated during further incubation. Based on these results, the maturation process of macrophages can be divided into at least the following four stages; glass‐nonadherent nonphagocytic precursor cells, glass‐adherent nonphagocytic precursor cells, immature macrophages with low phagocytic activity, and mature macrophages with full phagocytic activity. The addition of the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K) to cultures of spleen cells markedly suppressed the generation of macrophages. The suppressive effect of CPS‐K depended on its dosage, and the minimum concentration of CPS‐K showing a definite effect was 0.05 μg/ml. CPS‐K inhibited further generation of macrophages in either the nonadherent or adherent cell fraction at any time after the start of the culture. The suppressive effect of CPS‐K on the generation of macrophages could not be reversed by simple washing of spleen cells which had been kept in contact with CPS‐K for 3 hr. There was no evidence which showed that CPS‐K exhibited direct cytotoxic effects on spleen cells in the culture.
Effect of Polysaccharides from Ganoderma applanatum on Immune Responses
Authors: Nakashima Shoichi , Umeda Yukio , Kanada Taira
Published: November 2013

Prior intraperitoneal (i.p.) or oral administration of the polysaccharide preparation from a kind of mushroom, Ganoderma applanatum (Pers.) Pat. of Basidiomycetes, exerted an enhancing effect on the induction of delayed hypersensitivity (DH) to protein antigen as measured by the footpad reaction (FPR), and expanded the size of T cell memory for the IgG antibody response. One of the active principles was partially purified and found to be associated with a polysaccharide‐rich fraction. The induction of DH was enhanced by treatment with an appropriate dose of the mushroom extract, whereas increasing the dose resulted in almost complete loss of the enhancing activity.

The mechanism for the enhancing effect of the mushroom extract on the induction of DH was explored by the adoptive cell transfer technique. Although an i.p. injection of methylated bacterial α‐amylase (M‐BαA) in incomplete Freund's adjuvant (IFA) has been found to generate in the spleen the antigen‐specific suppressor T cells capable of inhibiting the induction of DH 5 days after immunization, the same treatment of mice given prior injections of the mushroom extract did not raise the suppressor cell activity, but transfer of these spleen cells (6 × 107) into syngeneic recipient mice which had been primed with a subcutaneous (s.c.) injection of M‐BαA in complete Freund's adjuvant (CFA) resulted in substantial amplification of the expression of DH. The absence of effector T cells for DH in the transferred spleen cells was confirmed by the failure to transfer DH into cyclophosphamide (CY)‐treated mice with the amplifying cells. The amplifying activity was antigen‐nonspecific and mediated by cells sensitive to treatment with anti‐θ antiserum plus complement. Therefore, the nonspecific enhancing effect of the mushroom extract could not be explained by the possibility that pretreatment with the extract eliminated the antigen‐specific suppressor T cells. Other adoptive cell transfer experiments revealed that nylon wool‐passed cells from mice unprimed but treated with the mushroom extract were able to exert an enhancing activity on the expression of effector T cells in DH. The results indicate that the treatment with an appropriate dose of the extract enhances the induction of DH by activation of the nonspecific amplifier T cells.
Partial Purification of Guinea Pig MIF by Affinity Column Chromatography Using Macrophages
Authors: Onozaki Kikuo , Haga Sinji , Miura Kaoru , Hashimoto Tatsuichiro
Published: November 2013

First we have confirmed the previous observation that the macrophage migration inhibitory factor (MIF) was adsorbed on normal peritoneal macrophages when they were incubated at 4 C for 60 min. It was found that macrophages fixed with 2% glutaraldehyde gave more reproducible results than viable cells in terms of “adsorption” of guinea pig MIF. The adsorption was achieved more completely at 37 C than at 4 C, indicating that this reaction is a temperature‐dependent phenomenon. Using these glutaraldehyde‐fixed macrophages, a kind of cell‐affinity column was successfully developed. The guinea pig MIF preparation lost its activity when it was passed through this affinity column, and MIF adsorbed on the column was recovered by elution with 0.1 M (L)‐fucose or 0.1 M (D)‐glucose. Such MIF active eluate was found to be at least 30–40 fold more pure than the original MIF preparation which had been previously fractionated according to its molecular weight. Therefore, this type of macrophage‐affinity column may be useful for the purification of MIF.
Interaction between 30 S Ribosomal Components in a Viomycin Resistant Mutant of Mycobacterium smegmatis
Authors: Mizuguchi Yasuo , Suga Kiyoko , Yamada Takeshi
Published: November 2013

A high level viomycin resistant mutant of Mycobacterium smegmatis ATCC 14468 (AC16) was analyzed genetically and biochemically in an attempt to understand the mechanisms of expression of high level viomycin resistance and co‐resistance to kanamycin and streptomycin. Genetic analysis has shown that at least three different genes (vicC, str, and kan) were involved in the phenotypic expression of drug resistance in AC16, and high level resistance to viomycin was due to interactions between the products of these genes.
Effect of Nucleosides on Interferon Production and Development of Antiviral State Induced by Poly I·Poly C
Authors: Machida Haruhiko , Kuninaka Akira , Yoshino Hiroshi
Published: November 2013

Effect of nucleosides both on induction of antiviral state in chick embryo cells (CEC) or rabbit kidney cells (RK13) and on interferon production in RK13 or mouse fibroblast cells (L cells) by polyriboinosinic‐polyribocytidylic acid (poly I·poly C) was studied. Addition of inosine or a fifty‐fifty mixture of inosine and uridine at a final concentration of 0.1 mM to 10 mM to a growth medium enhanced development of antiviral state in CEC. The nucleoside effect was also observed in RK13 at 0.1 mM but not at a concentration higher than 1 mM. Interferon production in RK13 by superinduction (sequential treatment with metabolic inhibitors after exposure to poly I·poly C) was enhanced 1.5‐ to 4.0‐fold by addition of the nucleoside mixture to the growth medium. When RK13 was pretreated with 10 units per ml of interferon and then superinduced by inhibitors, the enhancing effect of nucleosides on interferon production was not observed. Interferon production in L cells was potentiated a little by addition of 1 mM of the nucleoside mixture to the growth medium. The effect of nucleoside was not observed when the nucleosides were added after exposure to poly I·poly C. The nucleoside effect may be applicable for production of high titered interferon.
Effects of Tannic Acid and Its Related Compounds upon Chikungunya Virus
Authors: Konishi Eiji , Hotta Susumu
Published: November 2013

The present report describes not only the effects of tannic acid (TA; belonging to hydrolyzable tannins) and its related compounds upon the infectivity of Chikungunya virus (CHIKV) but also the mechanism involved in this phenomenon. Our data show that TA inactivates CHIKV in vitro. Since the inactivating effect turned out to be pH‐dependent and was suppressed by bovine serum albumin, it is most probable that the virus‐inactivating capacity of TA is attributable to its preferential binding to proteins of virus particles. Examination on the virus‐inactivating capacities of some TA‐related compounds and comparison of their structures indicated that the active site of TA and its analogues might be the phenolic hydroxyl groups in their molecules. It seems that the active groups interact with the proteins of virus particles, resulting in a reduction or loss of viral infectivity. Discussion is made on the specificity of the actions of tannins and the possibility of application thereof to chemicals which are useful to investigate the nature and properties of viral proteins.
Deletion Mutants from R Plasmids with Increased Copy Number
Authors: Ike Yasuyoshi , Hashimoto Hajime , Mitsuhashi Susumu
Published: November 2013

Rms 201–12 and Rms 201–46 are R mutants with increased copy number, and are derived from a conjugative plasmid Rms 201 that encodes resistance to five drugs, ampicillin (Ape), tetracycline (Tc), chloramphenicol (Cm), streptomycin (Sm), and sulfonamides (Sa). The mutants expressed increased levels of resistance to Ape, Sm, and Cm, and a decreased level of resistance to Tc than those of the parent Rms 201 plasmid.

When the Rms 201–12+ or Rms 201–46+ cells were inoculated onto plates containing a high concentration of Tc, colonies developed on the plate at a frequency of 10−3 to 10−4 after overnight incubation. The cells grown on the Tc plate carried a tet (Tc gene)‐deleted R mutant besides tet‐possessing Rms 201–12 or Rms 201–46, and we isolated the tet‐deleted R mutant by purifying the R+ cells on drug‐free plates. On the other hand, various deletion mutants possessing tet were isolated by prolonged culture of the cells. We have presented a circular gene order of Rms 201 by comparing the genetic markers of all deletion mutants derived from Rms 201, Rms 201–46, and Rms 201–12. The gene(s) regulating the copy number was closely linked to the rep gene. The gene(s) specifying entry exclusion was jointly lost with the tra region.
Cell Wall‐Bridge Maintaining Three Dimensional Structure of Cell Packets Formed by the Localized Suppression of Cell Separation of a Micrococcus lysodeikticus (luteus) Mutant1
Authors: Monodane Toshihiko , Matsushima Yoshio , Kotani Shozo
Published: November 2013

Cell packets of Micrococcus lysodeikticus (luteus) mutant strain MT grown in medium supplemented with trypsin consisted of a tetrad as the unit structure. An interstice was observed between the unit‐tetrads, and a three dimensional structure of cell packets was maintained by the cell wall‐bridge along the rim of the cell packets which linked each unit‐tetrad. This unique structure of strain MT cell packets seemed to occur when the cell separation was suppressed locally, i.e., when the cross wall inside the initial site of cell separation was cut off, while the wall outside the initial site of separation was not cut off but remained as a joint of the daughter cells.

The mechanism of cell wall‐bridge formation is discussed in connection with cell separation.
Growth and Sporulation of Auxotrophs of Bacillus subtilis in a Medium with Limited Nutrients
Authors: Nishihara Tsutomu
Published: November 2013

Growth and sporulation were examined for 30 auxotrophs of Bacillus subtilis in a chemically defined medium with suboptimal amounts of nutrients. All strains except for some adenine‐requiring mutants could not overtake sporulation stage II when amino acids, vitamins, or bases were limited, whereas they sporulated fairly well without limitation.

Abnormal structures, a cell with thickened cell wall and a cell with several refractile bodies, were found in some strains after the vegetative growth stopped.
The Physical State of Herpes Simplex Virus DNA in Infected Human Cells
Authors: Hirai Kanji
Published: November 2013

Treatment of herpes simplex virus type 1 (HSV‐1)‐infected human embryo lung (HEL) cells with phosphonoacetic acid (PAA) resulted in complete inhibition of HSV DNA replication. DNA was extracted from PAA‐treated HEL cells infected with HSV‐1 and centrifuged in a neutral CsCl density gradient. The HSV DNA sequences in the nuclei of PAA treated cells at 24 hr post infection banded at the same density as free HSV DNA (1.725 g/cm3), but a significant amount of viral DNA sequences were detected in the regions of cell DNA (1.700 g/cm3) as well as in the intermediate fractions as determined by hybridization with 3H HSV complementary RNA. The viral DNA sequences of lower density did not change in density by recentrifugation in a CsCl density gradient, but did change to the density of free viral DNA after treatment with EcoR1 restriction endonuclease. When the DNA from the nuclei of PAA treated cells was analyzed in an alkaline glycerol gradient, more than 95% of the viral DNA sequences were found in the free viral DNA fractions. Since the viral and cellular hybrid DNA represented approximately 33% of the total viral DNA sequences, it is concluded that some of the HSV DNA sequences in PAA treated, infected cells are associated with cell DNA by alkali‐labile bonds.
Ultrastructure and Partial Characterization of a Regular Array in the Cell Wall of Lactobacillus brevis
Authors: Masuda Kuniyoshi , Kawata Tomio
Published: November 2013

The cell wall of Lactobacillus brevis was revealed by electron microscopy to have an outer layer composed of a regular array. The morphological unit of the regular array appeared to consist of four spherical subunits, each about 2 nm in diameter, which were arranged in a tetragonal pattern about 4.5 by 7.0 nm in dimension. The regular array was composed of the tetragonal units in rows in two directions at an angle of about 75° to each other. The average spacing between the rows was about 10 nm in one direction and about 7 nm in the other. The tetragonally arranged subunits were removed from the cell wall by treatment with guanidine hydrochloride, urea, or sodium dodecyl sulfate (SDS) but not by the action of ethylenediaminetetraacetate, nonionic detergents, or proteolytic enzymes except pepsin. The regular subunits were shown to be composed of a protein with a molecular weight of about 51,000 by SDS‐polyacrylamide gel electrophoresis.
The Role of Macrophages in the Early Resistance to Mouse Hepatitis Virus Infection in Nude Mice
Authors: Tamura Teruko , Kai Chieko , Sakaguchi Akiko , Ishida Takuo , Fujiwara Kôsaku
Published: November 2013

Nude mice which had received intraperitoneal injection of silica simultaneously with infection of mouse hepatitis virus, NuU strain, died of severe necrotic hepatitis within 2 weeks postinfection, whereas those having received no silica survived for 3 weeks or more after challenge. Silica given day 4 postinoculation had no effect. The virus titers of the liver and spleen at day 4 as well as serum interferon levels at day 2 were much higher in silica‐treated mice than those without silica treatment. At day 2 or 3 postinoculation, silica‐treated mice were found to have a considerable number of necrotic foci in the liver with some neutrophil and lymphocyte infiltration, and viral antigen was present in the cytoplasm of some hepatocytes around necrotic foci. In contrast, those without silica treatment showed only some necrotic foci with some lymphocyte infiltration. Viral antigen was detected only in a few littoral cells but not in hepatocytes. The role of macrophages in the resistance at early stage of infection in nude mice is discussed.
The Activation Mechanism of Human Complement System by Immune Precipitate Formed with Rabbit IgG Antibody
Authors: Fujita Teizo
Published: November 2013

Immune precipitate (Ippt) formed between egg albumin and rabbit IgG antibody activated both pathways of the human complement system. On incubation with diluted serum, Ippt combined with several factors in the serum to form a complex which acquired C3‐ and C5‐cleaving activities. In serum chelated with ethyleneglycol tetraacetic acid (EGTA), C3‐ and C5‐cleaving activities of properdin system enzymes were formed on Ippt. Kinetic studies on the formation and the decay of C3‐ and C5‐cleaving enzymes on Ippt revealed that C3‐ and C5‐cleaving activities were almost dependent on the properdin system enzymes. The experiments in which C3‐cleaving activity formed on Ippt was inhibited by anti‐properdin or anti‐B but not by anti‐C4 supported the above results. The participation of the classical pathway was considered to accelerate the assembly of the properdin system enzymes.
Mycoplasma pulmonis Arthritis in Congenitally Athmic (Nude) Mice
Authors: Tanaka Hideo
Published: November 2013

Congenitally athymic BALB/cA nu/nu mice were employed to elucidate the role of the thymus in experimental Mycoplasma pulmonis strain m53 infection, and nu/+ mice were used for comparison. Chronic polyarthritis was frequently produced in both of nu/nu and nu/+ mice by intravenous injection of the organisms. Macroscopically, nu/nu mice developed severer arthritis and a much lower grade of resolution than nu/+ mice. Periarticular abscess, conjunctivitis, and emaciation were observed in some of the nu/nu mice, but not in the nu/+ mice. Mycoplasmas were isolated from joints and other tissues (including periarticular abscesses and eyelids) of infected nu/nu mice at higher frequencies as well as in greater quantities, and did not show any elimination trends for at least 20 weeks after inoculation. However, in nu/+ mice, mycoplasmas were almost exclusively located in joints, and distribution of organisms to the other organs disappeared soon after the infection. Increases in complement‐fixing antibody titers were not related to the inhibition of mycoplasmal spread. Thymus‐dependent functions that may in some way prevent growth and spread of mycoplasmas in mice are discussed.
“Pseudolysogenization” by RNA Phage Qβ
Authors: Watanabe Itaru , Sakurai Toshizo , Furuse Kohsuke , Ando Asako
Published: November 2013

We isolated fairly stable lysogenic‐like bacteria from a lysogenic state established between an amber mutant for the maturation protein gene of RNA phage Qβ (Qβam205) and its nonpermissive host BE110. These bacteria contained few mature phages intracellularly (less than 10−3 plaque forming unit per cell), continued to grow with a potentiality to produce Qβam205 spontaneously, and showed an immunity‐like response against homologous phage infection. These characteristics were maintained by growth in liquid medium containing anti‐Qβ serum. We designated these cells as pseudolysogenic bacteria. The relative amounts of RNA genomes in these pseudolysogenic cells (about 102 infectious RNA strands per cell) indicated that the RNA genomes could replicate in nonpermissive cells and be distributed in daughter cells synchronizing well with cell division.
Regulatory Mechanism of Reagin Production in Mice at the T Cell Level
Authors: Takatsu Kiyoshi , Tominaga Akira , Hamaoka Toshiyuki , Kitagawa Masayasu
Published: November 2013

Helper T cell activities specific for purified protein derivative (PPD) generated by immunization with Mycobacterium tuberculosis (Tbc) or PPD were investigated concerning adoptive IgE and IgG antibody responses. It is interesting that preferential triggering activity of IgG antibody response was observed when PPD‐reactive cells from mice immunized with Tbc were used as a helper cell source. The selective triggering of IgG B cells by Tbc‐primed cells was consistently observed using DNP‐primed B cell populations from mice immunized with DNP‐carrier conjugate in either ICFA or alum. T cell dependency of helper activity was demonstrated by the fact that treatment of Tbc‐primed cells with anti‐Thy 1 antiserum plus complement abolished their helper activity. We also demonstrated that purified T cell populations selectively triggerred IgG B cells.

Selective triggering of IgG B lymphocytes by Tbc‐primed T cells may not be due to the influence of suppressor T cells supposedly present in Tbc‐primed cells since this selectivity was not affected by X‐irradiation of Tbc‐primed T cell populations which may inactivate suppressor T cells. Furthermore, passive transfer of Tbc‐primed cells into normal recipient mice, the condition which may detect the suppressor T cell effect much more sensitively in IgE production, or preimmunization with Tbc 2 weeks before, did not suppress primary anti‐DNP IgE antibody response to DNP‐PPD. Thus, the observations presented here are favorable to the concept of the presence of IgG class‐specific helper T lymphocytes.

Furthermore, PPD‐reactive T cells from mice immunized with PPD itself exerted their helper function for triggering B cells of both IgE and IgG classes. This may also indicate that some of the components associated with Tbc other than PPD might negatively affect the development of PPD‐reactive helper T cells specific to the IgE class.

The generation of such IgG‐specific T cell activity in the presence of Tbc will be discussed in the light of the T cell population involved in the regulation of antibody responses of different immunoglobulin classes.
Antiplague Antibodies against Yersinia Pestis Fraction‐I Antigen in Serum from Rodents either Experimentally Infected or Captured in Harbor Areas of Japan, 1971–19771
Authors: Suzuki Sosuke , Hotta Susumu
Published: November 2013

Antiplague indirect hemagglutination tests were investigated with Yersinia pestis fraction‐I antigen and sera from experimentally infected rodents. The same tests with sera treated with acetone or acetone‐2‐mercaptoethanol were performed in parallel. Technical conditions were determined to insure the specificity and reproducibility of the reactions. Utilizing the methods studied, we carried out serological surveillance of plague with sera from wild rats (Rattus rattus and R. norvegicus) captured in the harbor areas of Japan during the period from 1971 to 1977. Four samples (all from R. norvegicus) out of 1,645 obtained in Kobe were positive. The public health significance of the data obtained is discussed.
Reactivity of Paul‐Bunnell Type Heterophile Antibody in Sera from Infectious Mononucleosis Patients with the Surface of Lymphoid Cells Carrying Epstein‐Barr Virus Genomes
Authors: Maeda Mitsuyoshi , Sairenji Takeshi , Hinuma Yorio
Published: November 2013

The possible presence of Paul‐Bunnell (PB) antigen on Epstein‐Barr virus (EBV)‐transformed lymphocytes was investigated. Of 23 EBV genome‐positive lymphoblastoid cell lines tested all but one absorbed PB type antibody from the serum of an infectious mononucleosis patient. The one EBV‐negative B cell line tested did not absorb the heterophile antibody. PB antibody, purified by an immunoadsorbent procedure using beef cell antigen, reacted with the EBV producer P3HR‐1 cell line in an indirect membrane immunofluorescence test and was shown to be IgM antibody. Titers of heterophile agglutinin and reactivity with the cell surface were reduced to the same degree by absorption with beef cell antigen but not with guinea pig kidney antigen. PB antibody was distinct from IgM antibody against the EBV‐determined membrane antigen, since the latter was not absorbed by beef cell antigen. PB antibody was also reactive with other EBV‐positive B cell lines (QIMR‐WIL, NC‐37, and Raji) which were free of surface IgM. No reaction occurred with the nonproducer P3HR‐1 line, a null cell line, and two T cell lines. The results suggest the presence of PB antigen on most EBV‐transformed B lymphocytes, and its appearance in each of the transformed lymphocytes of patients with acute infectious mononucleosis.
Hierarchy of T Cell Dependency in Antibody Response among Different Antigens
Authors: Amagai Takashi , Muramatsu Shigeru
Published: November 2013

T cell dependency of antibody response to polyvinylpyrrolidone (PVP), sheep red blood cells (SRBC), bovine gamma globulin (BGG), and bovine serum albumin (BSA) was examined. PVP and the other three are known as a T cell‐independent antigen and T cell‐dependent antigens respectively. Adult mice were thymectomized, X‐irradiated, reconstituted with syngeneic bone marrow cells (TxXB mice), with bone marrow cells plus thymus cells (TxXBT mice), or with bone marrow cells treated with anti‐Thy‐1.2 serum and complement (TxXB‐0 mice) and used as experimental animals. The anti‐PVP response of TxXBT mice was significantly lower than that of TxXB mice, suggesting that T cells exerted a suppressive effect on the response to PVP. Both IgM and IgG responses to SRBC and BGG occurred even in TxXB‐θ mice with the aid of bacterial lipopolysaccharide (LPS). However, a significant response to BSA was not observed in TxXB mice even in the presence of LPS or several other adjuvants. These results indicate that the T cell dependency of antigens is different among so called thymus‐dependent antigens, that antibody response less dependent on the helper action of T cells can be supported by LPS in the absence of T cells, and that anti‐BSA response seems to be extremely T cell dependent.
The Local Immune Response in the Mammary Gland of the Sow Following Infusion of a Protein Antigen
Authors: Bennell M.A. , Watson Dennis L.
Published: November 2013

A study was made of the local immune response in the udder of the sow following infusion of a soluble antigen. Four mammary glands of each of four pregnant sows were infused with ferritin prepartum. Samples of blood, colostrum, and milk were collected during the following lactation; animals were slaughtered and mammary tissue removed for immunohistology. Blood, colostrum, milk, and mammary tissues were similarly collected from nonimmunized (control) sows. Colostral and milk whey from immunized sows contained higher levels of immunoglobulins than whey from control sows. There was an increase in numbers of IgA‐containing plasma cells and total lymphoid cells in mammary tissue of immunized sows compared with controls. The results suggested that the local immune response was at least as great in non‐infused glands as infused glands of immunized sows.
Experimental Candida‐Induced Arteritis in Mice
Authors: Murata Hisao
Published: November 2013

An extract of Candida albicans isolated from a patient with typical mucocutaneous lymph node syndrome (MCLS) can produce coronary arteritis in a mouse when injected intraperitoneally. An unusual feature of this arteritis is that it is granulomatous, shows no fibrinoid change and is confined to the coronary arteries. These characteristics are quite similar to those found in patients with MCLS.
Cariogenic Virulence of Streptococcus mutans AHT Mutants with Altered In Vitro Adherence Abilities in Hamsters
Authors: Kishimoto Etsuo , Koga Toshihiko , Inoue Masakazu
Published: November 2013

Plaque‐forming ability and cariogenic virulence of Streptococcus mutans AHT mutants having varying in vitro adherence abilities were studied in hamsters reared on a high sucrose diet over a period of 64 days. The parent wild‐type strain produced heavy plaque and extensive carious lesions on molars. However, mutants M1, M2, and M13, which have lost the ability to form adhesive deposits on glass when grown in sucrose broth, and mutant M9, which adheres firmly to glass but in an extremely small amount, could not be recovered from the mouths of animals even after repeated inoculations at weekly intervals throughout the experimental period. Mutant M35, which produces a large amount of loosely adhering deposits in vitro, became established and persisted on molars, but its recovery was extremely low. None of the mutants, including M35, significantly produced plaque and caries in hamsters. Possible causes of this inability to form plaque and the avirulence of mutants are discussed.
Mode of Subacute Sclerosing Panencephalitis (SSPE) Virus Infection in Tissue Culture Cells
Authors: Ohuchi Masanobu , Ohuchi Reiko , Homma Morio
Published: November 2013

By the aid of freezing and thawing, cell‐free infectious virions were detected from an apparently nonproductive Vero cell line infected with Niigata‐1 strain of subacute sclerosing panencephalitis virus. The production of infectious virions was limited in amount and such virions were detectable only during a limited period after cell subculture. The infectious virions were filtrable through a 0.65μ membrane filter and neutralized completely by an antiserum against measles virus. The virions were banded at the density of 1.132, while Edmonston strain of measles virus banded at 1.164 in potassium tartrate density gradients. Infectious virions were also released from infected Vero cells by treatment of the cells in a hypotonic solution to an amount comparable to that obtained by freezing and thawing. Infection of normal culture of Vero cells with the infectious virions readily established a virus‐cell interaction identical to that in the original infected culture from which the virions were recovered.
Cell‐Mediated Immunity in Human Pregnancy
Authors: Fujisaki Syunichi , Mori Norimasa , Sasaki Toshio , Maeyama Masao
Published: November 2013

The function of thymus‐dependent lymphocytes (T lymphocytes) was studied in women during pregnancy and labor and postpartum by evaluating the blastogenesis of peripheral lymphocytes, which were stimulated with phytohemagglutinin‐P (PHA‐P) in both whole‐blood semimicroculture and purified lymphocyte culture. Data from 353 random samples (203 women) and 50 serial specimens from 10 women revealed that PHA‐P induced‐lymphocyte blastogenesis was significantly (p<0.005) reduced during pregnancy and labor but rapidly returned to normal several days after artificial termination in the early stage of pregnancy as well as after full‐term delivery. These results indicate that the T‐lymphocyte function in maternal peripheral blood is depressed by causes related to pregnancy. It seems very likely that depressed T‐lymphocyte function during pregnancy is caused by inhibitory factors in the blood plasma derived from the feto‐placental unit. Questions relating to the inhibitory factors in maternal plasma are discussed.
In Vitro Studies on Distribution of Indigenous Lactobacilli of the Gastrointestinal Tract of Rats
Authors: Watanabe Tsugio , Kawai Yasuo , Mutai Masahiko
Published: November 2013

To learn the biochemical mechanisms controlling the distribution of indigenous lactobacilli in the gastrointestinal tracts of rats, the effect of pH and stomach and cecal contents on lactobacillus distribution was investigated in vitro with a mixed culture of three lactobacillus strains isolated from the rat intestine. The pH of the growth medium affected the growth of lactobacilli strongly, irrespective of the lumenal contents. Lactobacillus fermentum outnumbered L. acidophilus and L. murini at low pH (pH 4.5; average pH of stomach contents of conventional rats) but at near neutral pH (pH 6.5; average pH of cecal contents of conventional rats), the growth of L. murini was predominant with all strains. More lactic acid was formed by lactobacilli in medium consisting of stomach contents than in cecal contents medium. L. murini grew in the nondialyzable fraction of the stomach contents and L. fermentum grew in the dialyzable fraction, but L. acidophilus did not grow in either fraction. L. murini grew in the nondialyzable fraction treated with hyaluronidase. In contrast, the nondialyzable fraction treated with pronase or chondroitinase did not allow L. murini to grow at all.
IgM Antibodies to Epstein‐Barr Virus‐Associated Membrane Antigen in Sera of Infectious Mononucleosis Patients
Authors: Harada Masataka , Sairenji Takeshi , Takaki Kazutaka , Hinuma Yorio
Published: November 2013

By the indirect immunofluorescence technique, IgM antibodies to the cell surface of an Epstein‐Barr virus (EBV) producer cell line, P3HR‐1, were detected in sera from infectious mononucleosis (IM) patients but not in sera from patients with Burkitt lymphoma or nasopharyngeal carcinoma nor in sera from healthy adult donors having antibodies to EBV‐specific viral capsid antigen (VCA). Titers of the IgM antibodies were higher in the earlier stages of IM, a pattern similar to that for IgM antibodies to VCA. The IgM antibodies to the cell surface were identified as being those against the EBV‐specific membrane antigen (MA) by the following criteria: (1) The antibodies were reactive to MA‐positive cell preparations but not to MA‐negative cell preparations. (2) Titers of the IgM antibodies were not significantly affected after absorption of sera with sheep red blood cells which could completely eliminate heterophil antibodies in the same sera. Detection of the IgM antibodies to MA may have a particular diagnostic value for providing evidence of a recent EBV infection.
Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response
Authors: Yokochi Takashi , Nakashima Izumi , Kato Nobuo
Published: November 2013

Changes in the number of cells and the weight of various lymphoid organs of mice, such as the regional lymph node (right inguinal node), spleen, thymus, bone marrow, and peripheral blood, were followed after the subcutaneous injection of the capsular polysaccharide of Klebsiella pneumoniae (CPS‐K). For comparison, the changes after injection of various polyclonal lymphocyte activators (PLA) including various preparations of bacterial lipopolysaccharide (LPS) were concurrently studied. The number of cells of all of the lymphoid organs tested and that of nucleated cells in the peripheral blood decreased significantly within a few days after injection of CPS‐K, and increased later. Above all, the increase in the number of cells and in the weight of the regional lymph node was most prominent (about 10 times larger than that of the normal control). Such a marked increase in the number of cells of the regional lymph node was not induced by the injection of any preparation of LPS or any other PLA tested. The initial decrease in the number of cells after CPS‐K injection was most marked and long lasting in the thymus. Although LPS prepared by Westphal's method from Escherichia coli O55 or Salmonella enteritidis exhibited a stronger decreasing effect on the number of cells of the thymus, the effect of LPS prepared by Westphal's method from E. coli O111 or that by Boivin's method from E. coli O55 was similar to that of CPS‐K. It is concluded therefore that CPS‐K has the ability to decrease the number of cells of various lymphoid organs, especially that of the thymus, initially after injection, which is a property in common with LPS, and CPS‐K has a unique ability to increase markedly the cells of various lymphoid organs, especially those of the regional lymph node, at later stages after injection. Considering that CPS‐K exhibits a much stronger adjuvant effect on the antibody response than does LPS or other polyclonal lymphocyte activators, it is suggested that this extraordinarily potent activity of CPS‐K in increasing the number of cells of the regional lymph node is closely related to its strong adjuvant action.
Properties of a Macrophage Cell Line Transformed by Simian Virus 40
Authors: Katsumoto Tetsuo , Takayama Hisao
Published: November 2013

A mouse macrophage clone (line NH‐1) transformed by simian virus 40 (SV40) was examined by electron microscopy. In the growing phase of the cultures, NH‐1 cells were non‐phagocytic and SV40 T antigen‐positive, and contained a large number of filament sheaths within their pseudopodia. In the late stationary phase, they became phagocytic, SV40 T antigen‐negative and contained a filamentous network within their psudopodia. In addition, NH‐1 cells in the late stationary phase were very similar to normal macrophages in other morphological properties.
Studies on the Sulfite Reduction Test for Clostridia
Authors: Kawabata Norihiko
Published: November 2013

Peptone‐yeast extract (PY) medium containing 0.035% ferric ammonium citrate as an indicator, 0.05% sulfite as a substrate, 0.05% cysteine as a reducer and 0.5% glucose was found to be suitable for observing the sulfite reduction test. The effect of added cysteine on the test was suppressed by the addition of glucose. In cultures of bacteria grown for 2 days at 37 C in medium containing the above ingredients, 121 among 132 strains of Clostridia, including 86 strains of Clostridium perfringens, gave a positive reaction. Although some strains of Salmonella and Proteus were positive, the specificity of the test for Clostridia was thought to be relatively high. Positive reactions in a resting cell system were limited to some species of Clostridia.
Reassembly of the Regularly Arranged Subunits in the Cell Wall of Lactobacillus brevis and Their Reattachment to Cell Walls
Authors: Masuda Kuniyoshi , Kawata Tomio
Published: November 2013

The reassembly of tetragonally arranged subunits in the cell wall of Lactobacillus brevis and the reattachment of the subunits to cell wall fragments were investigated by electron microscopy. The subunits dissociated from the cell wall with guanidine hydrochloride (GHCl) reassembled into the same regular array as seen in the native cell wall after dialysis against neutral buffer even in the absence of specific cations. The subunits could also reattach to the cell wall fragments from which they had been removed by treatment with GHCl, sodium dodecyl sulfate or cold trichloroacetic acid but not to those treated with hot formamide. Heterologous reattachment of the subunits occurred on cell wall fragments obtained from L. fermentum but not on those from L. plantarum or L. casei subsp. casei. On the basis of these observations and chemical analyses of the cell wall fragments, the subunits of L. brevis appeared to be bound by hydrogen bonds to a neutral polysaccharide moiety in the cell wall but not to peptidoglycan or teichoic acid.
Electron Microscopic Observation of New Transposable Elements Inserted into P22 Phage Genome from R Plasmids
Authors: Suzuki Kenji , Mise Katsutoshi , Nakaya Rintaro
Published: November 2013

By using phage P22spl, a deletion mutant of phage P22, the structures of two new transposons on P22 genomes were studied by the electron microscopic heteroduplex method. One of these was the Cm (chloramphenicol) transposon derived from an R plasmid, NR1, and the other the Km (kanamycin) transposcn from pNR502. The heteroduplex between P22 phage DNAs with and without the Cm transposon revealed that the Cm transposon was similar in structure to the Tn9 element, a well‐known Cm transposon derived from the R plasmid pMS14. On the other hand, the Km transposon of pNR502 was quite different in structure from other Km transposons reported previously. This transposon consists of a 6.8 kilo‐base (kb) segment of DNA, in which a short inverted repeat is contained. The heteroduplex experiments showed that a 4.5 kb segment of DNA was deleted from the P22 genome in the P22spl genome. Because of a shorter unit length of the genome, phage P22spl is considered to be useful for assaying various kinds of transposable elements.
In Vitro Proliferative Activity of Spleen Cells in Mice Infected with Attenuated Salmonella typhimurium
Authors: Saito‐Taki Tatsuo , Asou Hiroaki , Nakano Masayasu
Published: November 2013

Proliferative activity of cultured spleen cells obtained from mice 1 to 5 weeks after infection with attenuated strains of Salmonella typhimurium was examined in the presence or absence of lipopolysaccharide (LPS) or concanavalin A (Con A). Spontaneous uptake of 3H‐thymidine (TdR) by cells taken from infected mice at the 2nd and 3rd weeks was obviously lower than that by cells from uninfected, control mice. Cells from infected mice at the 4th and 5th weeks also showed a lower proliferative response to LPS than that of the controls. However, the responses of the cells to Con A remained virtually unchanged during the entire period. Furthermore, the reduction of spontaneous 3H‐TdR uptake by the cells could be achieved also by the injection of heat‐killed instead of living organisms.

The T‐ and B‐lymphocyte populations of these spleen cells were examined by the dye exclusion cytotoxic test using rabbit anti‐mouse T‐ and anti‐mouse B‐lymphocyte sera, respectively. There was some alteration of the populations in the cells, but it did not correlate with the reduction in 3H‐TdR uptake.

Results of expriments with cultured cells reconstituted with lymphocytes and macrophages isolated from spleen cells suggested that the spontaneous reduction of proliferative activity observed in cells taken from the infected mice could be attributed to the dysfunction of macrophages.
Superoxide Anion‐Generating Activities of Macrophages as Studied by Using Cytochalasin E and Lectins as Synergistic Stimulants for Superoxide Release
Authors: Kayashima Kohji , Onoue Kaoru , Nakagawara Akira , Minakami Shigeki
Published: November 2013

Treatment of macrophages with cytochalasin E in combination with a lectin was found to stimulate the generation of superoxide anions (O2‐) very efficiently. The macrophages stimulated with concanavalin A, phytohemagglutinin or wheat germ agglutinin released superoxide, but cells pretreated with cytochalasin E released much greater amounts of superoxide, without notable lag time, upon stimulation with the lectin. Wheat germ agglutinin was found to be the most efficient stimulant among the lectins tested. Superoxide generation in guinea pig macrophages was shown to be dependent largely on cytoplasmic glucose metabolism and to some extent on mitochondrial respiration, since the superoxide release was largely but not totally inhibited by 2‐deoxyglucose and to a lesser extent by antimycin A or KCN. The method presented is sensitive and allows rapid assay of the superoxide‐generating activity with only 1–5 × 105 macrophages for a single determination. In application of this technique, elevation of the superoxide‐generating activity was shown with macrophages elicited by chemical inflammation or those obtained from mice after treatment with tubercle bacilli.
Gene Expression of Ampicillin Resistance Transposons, Tn2601 and Tn2602
Authors: Yamamoto Tomoko , Katoh Rieko , Shimazu Akemi , Yamagishi Saburo
Published: November 2013

To establish the mode of gene expression specified by transposon, we investigated the correlation among the homology of the DNA sequence, the extent of transposon‐specific transcription, the specific activity of penicillinase (PCase) per cell, and the transposition frequency by using two ampicillin resistance transposons (TnAs), Tn2601 and Tn2602. Although both the TnAs specify the so‐called type I PCase, Tn2602 always conferred 10–to 20‐fold higher PCase activity per cell than Tn2601 regardless of the kind of replicon carrying TnA. The transposition frequency of Tn2602 also was 8 to 50 times higher than that of Tn2601 in all combinations of donor and recipient plasmids examined. As a result, the transposability expressed by the TnA was thought to correlate with the productivity of PCase in the cell specified by the corresponding TnA. The level of TnA‐specific transcription of Tn2602 was noticeably higher than that of Tn2601, whereas the two TnAs shared a high degree (more than 90%) of DNA sequence homology. These results suggest that the difference in rates of transcription of the two transposons plays a key role in determining the difference in the productivity of PCase and the transposition‐protein(s) of Tn2601 and Tn2602.
Location of Elements in Ashed Spores of Bacillus megaterium
Authors: Nishihara Tsutomu , Ighikawa Tomio , Kondo Masaomi
Published: November 2013

The structure of the skeleton of spores of Bacillus megaterium was examined after ashing in a plasma asher and the elemental composition of the ashed whole spores was determined with an analytical electron microscope. All spores were ashed in situ although they shrank by about 15%. Even P and S, in addition to metals, were recovered well from ashed samples. Ash was rich in the core and the coat, and poor in the cortex. Ca, P, S, and Mg were detected in the core and coat of the spore of B. megaterium QM B1551. Ca in the core was markedly decreased by germination or autoclaving. In the spore of B. megaterium ATCC 19213, almost all of the ash was detected in the core and its elemental composition was similar to that of the core of the strain QM B1551 spore.

These results suggest strongly that the core is the site of Ca associated with dipicolinic acid.
Coliphage HK243: Biological and Physicochemical Characteristics
Authors: Dhillon Tarlochan S. , Dhillon Elvera K.S. , Toyama Sakuji , Linn Stuart
Published: November 2013

Coliphage HK243 can form plaques on Escherichia coli C and K‐12, but not B. The plaques are 1–2 mm in diameter and are opaque areas which clear upon exposure to chloroform vapor. During one‐step growth, the eclipse and the latent periods are 20 and 30 min, respectively. Phage‐infected cells continue to produce cell‐free plaque‐forming units for as long as 80 min after the end of the latent period, although at high multiplicities of infection (MOI) most cells lyse. No lysogenic bacteria have been found among survivors, so HK243 is considered a virulent phage. Some of the cells surviving a high MOI challenge are maltose negative and resistant to both HK243 and coliphage lambda. This fact has made possible the isolation of lambda‐resistant mutants of lambda‐lysogens. However, no serological cross‐reaction between the phages lambda and HK243 has been detected. Genetic data involving three essential loci and a locus controlling plaque morphology suggest a circular linkage map. The virions are tadpole‐shaped with an icosahedral head 68 nm long which is attached to a flexible tail 131 nm long. The phage has a linear, duplex DNA genome of molecular weight approximately 44 × 106 and a base composition of 33% adenine, 31% thymine, 16% guanine, and 20% cytosine.