bluuebunny/crossref_metadata_2025 huggingface dataset:
Sex‐ and Age‐Specific Prevalences of HBeAg and Anti‐HBe among HBsAg Carriers with or without Liver Function Abnormalities in Okinawa, Japan
Authors: Hayashi Jun , Kashiwagi Seizaburo , Ikematsu Hideyuki , Nomura Hideyuki , Kajiyama Wataru , Noguchi Akinori , Ikeda Kiyoshi
Published: November 2013

We investigated sex‐ and age‐specific prevalences of hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti‐HBe) in 1,163 carriers of hepatitis B surface antigen (HBsAg) in Okinawa, Japan, and followed them up for longer than one year in correlation with liver function abnormalities. Elevated serum transaminase levels were found in 160 (13.8%) of them. The prevalence of liver function abnormalities was significantly higher in male carriers (127/690, 18.4%) than in female carriers (33/473, 7.0%) at a P value of 0.001. In asymptomatic carriers, the prevalence of HBeAg was 13.2% and that of anti‐HBe 80.0%, significantly different from 41.9% and 54.4%, respectively, in carriers with elevated transaminase levels (P<0.001). In asymptomatic carriers, the mean age of HBeAg‐positive carriers was 16.7 years which was much lower than the 22.8 years in carriers with liver function abnormalities. The prevalence of elevated transaminase levels was significantly higher in carriers with HBeAg than in those with anti‐HBe (33.8% vs. 9.7%, P<0.001). Based on these results, the prolonged positivity for serum HBeAg would qualify as a predictor for deteriorating liver function among HBsAg carriers.
Selective Inhibition of High‐ but Not Low‐Affinity Interleukin 2 Binding by Lectins and Anti‐Interleukin 2 Receptor α Antibody
Authors: Fujii Masahiro , Sugamura Kazuo , Nakamura Masataka , Ishii Takehisa , Hinuma Yorio
Published: November 2013

The present study demonstrated that various reagents can specifically reduce the affinity of high‐affinity interleukin 2 receptor (IL‐2R) but not that of low‐affinity IL‐2R. They included lectins such as wheat germ agglutinin (WGA), concanavalin A and phytohemagglutinin, and a chemical cross‐linker, glutaraldehyde, in addition to anti‐IL‐2R monoclonal antibodies, HIEI and H‐47. The affinity of the high‐affinity IL‐2R was reduced when the cells were treated with WGA or H‐47 before, but not after, addition of 125I‐labeled interleukin 2 (IL‐2). Their inhibitory effects were also demonstrated by the chemical cross‐linking method. On treatment with the reagents, the IL‐2 binding to both IL‐2Rα and β chains was inhibited and these inhibitory effects were seen only when the reagents were added before IL‐2 addition, like their high‐affinity reducing effects. These results support a supposition that the high affinity IL‐2R is generated by assembly of the α and β chains, and suggest that the IL‐2 binding to IL‐2Rα and β chains could induce stable constitution of the high‐affinity state of IL‐2R, but these affinity modulating reagents could perturb the optimum association between α and β chains to generate the high‐affinity IL‐2R.
Immortalization of Murine Leukocytes by Oncogenes
Authors: Komatsubara Shin , Mori Kazuyasu , Hatanaka Masakazu , Amagai Takashi , Inaba Kayo , Shimizu Satoshi , Muramatsu Shigeru
Published: November 2013

In the course of study to obtain murine dendritic cell lines using oncogenic retroviruses, we have established several immortalized cell lines with characteristics different from those of dendritic cells. The transformants were mainly round nonadherent cells, capable of growing in soft agar, and negative for nonspecific esterase activity. Profiles of cell surface antigens were examined by indirect immunofluorescence technique. The cell lines were positive for Fc receptor (2.4G2), J11d (J11d.2), and B220 (RA3‐3A1/6.1) antigens and negative (or dull positive in small percentages) for Ia (M5/144.15.2), IL‐2 receptor (3C7), Thy‐1 (B5‐5), Mac‐1 (M1/70.‐15.11.5), and macrophage (F4/80) antigens. They were negative for both surface and cytoplasmic immunoglobulins. Several clones were established from these transformant cell lines and cell surface antigens were examined. Antigenic profiles of these clones were very similar to those of the parental cell lines. Some of these clones, however, seemed to increase their Ia antigen expression. The results suggest that the transformants originated from early B‐lineage cells.
Primary Role of NADH Formed by Glucose Dehydrogenase in ATP Provision at the Early Stage of Spore Germination in Bacillus megaterium QM B1551
Authors: Sano Keiji , Otani Mieko , Uehara Reiko , Kimura Masahiro , Umezawa Chisae
Published: November 2013

Metabolic events involved in energy metabolism were studied in order to evaluate the ATP‐forming ability of Bacillus megaterium QM B1551 spores at the very early stage of germination. When heat‐activated spores were germinated on glucose as a sole substrate, its oxidation into gluconate (catalyzed by glucose dehydrogenase, EC 1.1.1.47), the accompanying NADH formation, oxygen uptake, and RNA synthesis were initiated immediately after germination, even when anaerobic breakdown of 3‐phosphoglycerate (an ATP source for spores) and the subsequent glucose metabolism via the phosphorylating pathway were impaired by potassium fluoride (KF). In contrast, fructose metabolism and the accompanying metabolic events did not begin until a few minutes after triggering of germination, and those events were entirely abolished by KF, indicating that fructose metabolism is initiated exclusively via its phosphorylation by the ATP derived from endogenous 3‐phosphoglycerate. Thus those results provided further evidence for our previous proposal (Otani et al (1987) Microbiol. Immunol. 31: 967–974; Sano et al (1988) Biochem. Biophys. Res. Commun. 151: 48–52) that the first molecules of ATP in germinating spores can be efficiently generated via aerobic oxidation of NADH, which is formed by glucose dehydrogenase. Fluorescence monitoring of NADH in germinating spores also supported this conclusion.
Experimental Chronic Pulmonary Infection in Mice Caused by Klebsiella pneumoniae
Authors: Iizawa Yuji , Nishi Takeshi , Kondo Masahiro , Imada Akira
Published: November 2013

Examination of mouse strain differences in susceptibility to experimental respiratory tract infection with Klebsiella pneumoniae 27 revealed that a chronic pulmonary infection model could be established using CBA/J mice. After 6 × 105 colony‐forming units of K. pneumoniae 27 were inoculated into the lung, the bacterial counts in the lungs changed with time showing four different phases: initial decrease, regrowth, steady‐state, and final increase leading to death. Throughout the course of the infection, the challenge bacteria were isolated mainly from the respiratory organs. Pulmonary gross lesions appeared on day 2 after infection and persisted thereafter. Lobar consolidation was the primary lesion and occurred mainly in the anterior and middle lobes of the right lung, and the median lobe. Mice began to die from 4 weeks after aerosol exposure. This model may be useful for investigating the pathogenesis of chronic pulmonary infection by Klebsiella and its therapy.
Effect of Recombinant Murine Interferon‐Beta on the Replication of Murine Cytomegalovirus
Authors: Nakamura Kazuo , Eizuru Yoshito , Minamishima Yoichi
Published: November 2013

Pretreatment of mouse embryo fibroblasts (MEF) with recombinant murine interferon‐β (rMuIFN‐β) induced a high level of intracellular 2′,5′‐oligoadenylate (2–5A) synthetase activity. However, murine cytomegalovirus (MCMV) replicated under such condition, indicating that MCMV is relatively insensitive in vitro to rMuIFN‐β. Thus, there was a dissociation of 2–5A synthetase activity and antiviral activity against MCMV. In contrast to MCMV, the two parameters were closely associated in the case of vesicular stomatitis virus (VSV).
Effect of Activation of the Epstein‐Barr Virus Genome on Expression of B Cell Differentiation Antigens of Burkitt's Lymphoma Lines
Authors: Yokochi Takashi , Inoue Yoshiko , Iwata Hitoshi , Miyadai Toshiaki , Kimura Yoshinobu
Published: November 2013

Using anti‐human B cell monoclonal antibodies prepared against B1 (CD20), B2 (CD21), B4 (CD19), and BB‐1 (B lymphoblast antigen‐1), we compared the expression of B cell differentiation antigens on a Jijoye‐P3HR‐1 cell line family of Burkitt's lymphomas. The expression of BB‐1 and B2 antigens was faint on P3HR‐1 K cell line which is an Epstein‐Barr virus (EBV) high producer. On the other hand, B1 and B4 antigens were strongly expressed on it. It was also found that BB‐1 expression decreased on P3HR‐1 cells after activation of intracellular EBV genes by treating chemically with tumor‐promoting agent (TPA) and n‐butyrate, or on Raji cells on superinfection with EBV. This decrease of BB‐1 was blocked by the additional treatment with retinoic acid, an inhibitor of virus replication. Dual immunofluorescence staining analysis showed that the individual cell expressing EBV‐associated antigens expressed BB‐1 antigen only marginally. The relationship between the change in phenotypes of host B cells and the activation of the EBV genome is discussed.
Immunoelectron Microscopic Studies on Spore Coat Proteins of Bacillus megaterium
Authors: Imagawa Masayoshi , Ohtsuka Mikiko , Nakatani Yoshihiro , Oku Yuichi , El‐Belbasi Hussein I. , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

An immunochemical staining technique for the spore coat proteins of Bacillus megaterium ATCC 12872 was developed using colloidal gold as a second antibody. For reducing the non‐specific immunogold binding and increasing the specific binding, the affinity‐purified IgG was used as a first antibody. In sporulating cells at t10, gold particles were found not only in the spore coat but also in the mother cell cytoplasm, suggesting that some coat proteins were synthesized in the cytoplasm. Use of the specific affinity‐purified antibody to 48K‐protein demonstrated that this protein was one of the components of the outer coat.
Role of the Protease in the Permeability Enhancement by Vibrio vulnificus
Authors: Miyoshi Shin‐ichi , Shinoda Sumio
Published: November 2013

The protease produced by Vibrio vulnificus enhances vascular permeability through histamine release from mast cells and activation of the plasma kallikrein‐kinin system which generates bradykinin when injected into the dorsal skin. V. vulnificus living cells also enhanced vascular permeability within a few hours after the injection into the dorsal skin. The permeability‐enhancing activity of living cells was greatly reduced by addition of soybean trypsin inhibitor, a specific inhibitor for plasma kallikrein‐kinin system, or anti‐protease IgG. Two protease‐deficient mutants induced by nitrosoguanidine treatment had only one‐tenth permeability‐enhancing activity of a wild‐type strain. These results indicate that V. vulnificus elaborates the protease in vivo and that the protease elaborated enhances vascular permeability through release of chemical mediators such as histamine and bradykinin and forms edema.
Induction of Murine B Cell Proliferation and Immunoglobulin Synthesis by Some Bacterial Ribosomes
Authors: Lafont Sergyl , Millet Isabelle , Kouassi Edouard , Jeannin Michele , Revillard Jean‐pierre , Normier Gerard , d'Hinterland Lucien Dussourd
Published: November 2013

Ribosomal preparations from Klebsiella pneumoniae, Haemophilus influenzae, Streptococcus pyogenes, and Streptococcus pneumoniae were investigated with respect to their activating capacity towards murine lymphoid cells. The proliferation of BALB/c spleen cells was induced in a dose‐dependent fashion (from 1 to 100 μg/ml) by ribosomes of K. pneumoniae, H. influenzae, and S. pyogenes with a peak activity at 48 or 72 hr of culture. The majority of the blast cells induced by these ribosomal preparations were positive for surface‐immunoglobulin (S‐Ig) and negative for Thy 1.2. Furthermore, K. pneumoniae, H. influenzae, and S. pyogenes ribosomes induced the synthesis of IgM and some IgA. Cell proliferation and induction of IgM production were also demonstrated with the 3 ribosomal preparations using spleen cells from athymic nude (nu+/nu+) mice, Lyb‐5‐defective CBA/N spleen cells, B cell‐enriched and T cell‐depleted BALB/c spleen cell suspensions, as well as spleen cells from the Ips gene‐deficient C3H/HeJ strain. Cell culture supernatants contained specific anti‐ribosome IgM antibodies. Antibodies of other specificities (anti‐sheep erythrocytes) were also demonstrated in supernatants from K. pneumoniae‐stimulated cultures. Evidence against a possible role of contamination of K. pneumoniae and H. influenzae ribosomes by lipopolysaccharide‐ or lipid A‐associated proteins in this effect is discussed. Ribosomes from S. pneumoniae did not induce 3H‐thymidine incorporation nor Ig production. None of the 4 ribosomal preparations was found to stimulate T cell blastogenesis or to induce interleukin‐2 production by naive BALB/c spleen cells. Finally, ribosomes from H. influenzae, S. pyogenes, S. pneumoniae but not those of K. pneumoniae stimulated interleukin‐1 production by adherent spleen cells, from BALB/c mice.
Dendritic Cell Activating Factor Produced by Mouse Macrophage Hybridoma
Authors: Ito Akira , Woo Hee Jong , Imai Yasuyuki , Osawa Toshiaki
Published: November 2013

A mouse macrophage (Mφ) hybridoma which produces a soluble factor responsible for the cooperation between Mφ and spleen dendritic cells (DC) was constructed. The antigen‐presenting activity and the stimulator cell activity in the allogeneic or syngeneic mixed leukocyte reaction of DC were significantly augmented when DC were incubated with the culture supernatant of the hybridoma treated with various stimulants including latex beads. The monokine present in the culture supernatant of the hybridoma, called dendritic cell‐activating factor (DCAF), augmented the production of lymphocyte‐activating factor by DC while Ia expression of DCAF‐treated DC was not altered. DCAF had no effect on the antigen‐presenting activity of peritoneal resident Mφ or B cell blasts while the antigen‐presenting activity of spleen Mφ was enhanced, but the degree of the enhancement was much less than that of spleen DC. DCAF was found to have the following properties: its pI value is between pH 4 and 5; it is stable at pH 2 to 10; and it loses its activity on incubation at 75 C for 30 min. When the culture supernatant of the hybridoma stimulated with latex beads was subjected to gel filtration, the DCAF activity was detected in the 20 Kd to 25 Kd, 30 Kd to 40 Kd, and 50 Kd to 60 Kd molecular weight regions. The 30 Kd to 40 Kd fraction, which is the major peak fraction, was further purified by ion‐exchange chromatography followed by gel‐filtration chromatography. When each fraction was subjected to SDS‐PAGE, a 30 Kd band corresponding to the DCAF activity was observed and DCAF was purified to about 90% purity.
Arachidonic Acid Release Is Closely Related to the Fcγ Receptor‐Mediated Superoxide Generation in Macrophages
Authors: Ida Eiichi , Sakata Atsuko , Tominaga Mari , Yamasaki Hisato , Onoue Kaoru
Published: November 2013

Stimulation of macrophages with IgG2 immune complexes induced dose‐dependently the O2− generation and the release of arachidonic acid and its metabolites. This FcγR‐mediated O2− generation was inhibited by a phospholipase A2 inhibitor, 4‐p‐bromophenacyl bromide (4‐pBPB), in parallel to the dose‐dependent inhibition of arachidonic acid release. The main arachidonic acid metabolites released were shown to be prostaglandin E2 and thromboxane B2 and blocking of the production of these metabolites by indomethacin did not inhibit the O2− generation. Inhibition of the FcγR‐mediated O2− generation and the arachidonic acid release by the C‐kinase inhibitor, 1‐(5‐isoquinolinesulfonyl)‐2‐methylpiperazine (H‐7), was less intense than by 4‐pBPB. These results support the previously proposed hypothesis that arachidonic acid acts as an intracellular activator of the FcγR‐mediated O2− generation in macrophages. Although the C‐kinase activation may also contribute to the activation of the O2−‐generating system, arachidonic acid release appears to play a major role in FcγR‐mediated O2− generation. In contrast, activation of C‐kinase seems to be contributing mainly in the induction of both the arachidonic acid release and O2− generation by 12‐o‐tetradecanoylphorbol 13‐acetate (TPA). Furthermore, suboptimal concentrations of TPA and arachidonate were found to act synergistically to stimulate O2− generation and the inhibition study suggested a positive synergism between C‐kinase and arachidonic acid release to induce O2− generation. This synergistic action may have general importance in receptor‐mediated O2− generation.
Characterization of Canine Distemper Viruses Adapted to Human Neural Cells
Authors: Morikawa Yuko , Yoshikawa Yasuhiro , Yamanouchi Kazuya
Published: November 2013

The biochemical characteristics of canine distemper virus (CDV) adapted to three human neural cells (glioblastoma, oligodendroglioma, and neuroblastoma cells) were compared with those of the unadapted original virus. The specific gravity of the virions and nucleocapsids of the original and the three adapted viruses were not different. The molecular weights of genomic RNA and messenger RNAs encoding H, F, P, and NP proteins of the adapted viruses as estimated by Northern blot hybridization were similar to those of the original virus. By T1‐resistant oligonucleotide analysis of the genomic RNA, the glioblastoma‐ and the neuroblastoma‐adapted viruses gave two more spots than the original virus; the oligodendroglioma‐adapted virus had a pattern identical to that of the original virus. By two‐dimensional gel electrophoresis of virion proteins, we found a difference in the isoelectric point of the viral envelope proteins H and F between the original and the adapted viruses. These results suggest that viral genomic changes occurred during adaptation, resulting in the alteration of viral envelope proteins.
Ecological Studies on Reovirus Pollution of Rivers in Toyama Prefecture
Authors: Matsuura Kumiko , Ishikura Mitsuhiro , Nakayama Takashi , Hasegawa Sumiyo , Morita Osayuki , Uetake Hisao
Published: November 2013

Reoviruses (reos) were isolated from river water in various areas of Toyama Prefecture. The frequency of reo isolation was higher in the river water, the basin of which has a larger human population. The degree of river contamination with reo paralleled that with the Escherichia coli group of organisms, and reos were frequently isolated from sewage, too. The high antibody‐positive (≥ 1: 8 or ≥ 1: 10) rates against reos in humans and other animals tested (swine, cattle, and field rodents) indicated their wide‐spread infection with reos. These results suggested that the major source of reos present in the river water may be the excretion by humans and other animals, especially the former. Survival experiments in which reos were added into the filtered or centrifuged river water and kept at various temperatures, revealed that reos survived for more than 3 years at 5 C. In the field experiment where reos suspended in cellophane tubes were kept in an agricultural water stream in winter (water temperatures below 10 C), they survived for 6 months until the water temperature rose above 20 C in summer.
Protective Immunity Induced by Porin in Experimental Mouse Salmonellosis
Authors: Matsui Katsuhiko , Arai Toshihiko
Published: November 2013

The induction of protective immunity to mouse salmonellosis by porin from Salmonella typhimurium LT2 was studied. The immunization with porin induced a high level of protective immunity to salmonellosis in BALB/c mice. Mice immunized with porin exhibited significant levels of delayed‐type hypersensitivity response and interleukin‐2 production, indicating that porin was capable of inducing cell‐mediated immunity (CMI). Furthermore, we found that both T cells and sera taken from the porin‐immunized mice could transfer the protection against salmonellosis into nonimmunized mice. These observations suggested that a high level of the protection to salmonellosis obtained by the porin immunization resulted from the induction of CMI in addition to humoral immunity.
Effects of Infection by HIV‐1, Cytomegalovirus, and Human Measles Virus on Cultured Human Thymic Epithelial Cells
Authors: Numazaki Kei , Goldman Hy , Bai Xin‐Qing , Wong Inés , Wainberg Mark A.
Published: November 2013

A tissue culture system for the growth of human fetal and infantile thymic epithelial (TE) cells has been established and characterized. We have investigated the effects of infection of these cells by human cytomegalovirus (CMV), measles virus, and human immunodeficiency virus type‐1 (HIV‐1). In the case of CMV, morphological changes were apparent by 2–4 days after viral inoculation of infantile TE cells. CMV‐related antigens were detected by immunofluorescence after 12 days, and progeny infectious CMV was recovered from culture media after 18 days. Following infection by measles virus, distinctive, multinucleated giant TE cells appeared in both cultures of fetal and infantile TE cells. Measles virus‐inoculated TE cells displayed an altered phenotype, as revealed by reaction with monoclonal antibodies with specificity for a variety of TE markers. Finally, infection of TE cells by HIV‐1 resulted in cellular disarrangement, increased numbers of Hassall's corpuscles, and multinucleated giant cells. An increase in the number of cells reactive with monoclonal antibodies, specific for Hassall's corpuscles, was observed in the case of cells infected by either measles virus or HIV‐1. These findings suggest that a variety of different viruses can successfully infect thymic epithelial tissue. Because of the important role of the thymus in development of the immune system, it is reasonable to conclude that viral infection of thymic tissue might play an important role in virus‐mediated suppression of immune responsiveness.
Endotoxic Properties of Chemically Synthesized Lipid A Analogs
Authors: Yoshida Masao , Hirata Michimasa , Inada Katsuya , Tsunoda Nobuko , Kirikae Teruo , Onodera Tsuyoshi , Ishikawa Yoshihito , Sasaki Osamu , Shiba Tetsuo , Kusumoto Shoichi , Galanos Chris , Lüderitz Otto , Kondo Seiichi , Hisatsune Kazuhito
Published: November 2013

Biological activities of two groups of synthesized lipid A analogs, the counterpart of biosynthetic precursor, Lehmann's Ia type, 406, and E. coli lipid A type, 506, as well as their non‐phosphorylated, and mono‐phosphorylated analogs were investigated. The activities employed included four bone marrow cell reactions in mice, mice skin reaction, leukocytes migration in rabbits' cornea, and hemagglutination. Compound 406 and 506 elicited bone marrow reactions in mice and hemagglutination of mouse RBC, although 406 failed to elicit hemorrhage and necrosis also in mice skin. Compound 406 did not elicit corneal reaction in rabbits. The results suggest that for elicitation of this reaction and mice skin reaction, acyloxyacyl structure is required. Cytotoxicity and thromboplastin production of four bone marrow reactions had been reported by us to be endotoxic reactions, since these had not been elicited by peptidoglycan of Lactobacillus and Staphylococcus (1981) and 300 series synthetized analogs (1984) which did not have endotoxic structures. From these results, it seems that these two marrow reactions and hemagglutination require, as does the limulus test, the lipid A part structure as is present in 406.
A Proposed Life Cycle Model of Spiroplasma mirum Based on Scanning Electron Microscopical Observations of Growth in Liquid Culture
Authors: Itoh Kikuji , Pan In‐Jen , Koshimizu Kaoru
Published: November 2013

Cells of Spiroplasma mirum strain SMCA were grown in PPG broth and examined by scanning electron microscopy. Comparison of the results at different time intervals allowed for a model of the life cycle of S. mirum to be proposed. Under favorable growth conditions, helical filament formation was initially observed, followed by the formation of small spherical structures originating from each filament. In old culture, large spherical bodies appeared from entangled helical filaments. From the larger spherical bodies, granular bodies representing the smallest reproductive units were produced to continue the life cycle.
Comparative Characterization of Spirosomes Isolated from Lactobacillus brevis, Lactobacillus fermentum, and Lactobacillus buchneri
Authors: Nomura Shuichi , Masuda Kuniyoshi , Kawata Tomio
Published: November 2013

Spirosomes, cytoplasmic fine spirals, were isolated and purified from Lactobacillus brevis ATCC 8287, L. fermentum F‐1, and L. buchneri ATCC 4005, and their morphological, biochemical, and immunological properties were investigated. The spirosomes of these lactobacilli were morphologically indistinguishable from one another, and they had the same buoyant density of 1.320 g/cm3 in CsCl. All of the spirosomes were composed of a single protein, spirosin, with an apparent molecular weight of about 95,000 for L. brevis and L. fermentum and of about 96,000 for L. buchneri as estimated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). The spirosins from the three lactobacilli were compared by peptide mapping on SDS‐PAGE after cleavage with N‐chlorosuccinimide and limited proteolysis with Staphylococcus aureus V8 protease. The peptide map of the L. brevis spirosin was identical with that of the L. fermentum spirosin, whereas it was markedly different from the L. buchneri spirosin. The amino acid composition of the L. brevis spirosin was almost similar to that of the L. fermentum spirosin, while it differed appreciably from the L. buchneri spirosin. Using antiserum against the L. brevis spirosin, immunodiffusion test revealed that the antigenicity of the spirosomes from L. brevis was identical with that from L. fermentum, whereas it was partially different from that from L. buchneri.
Effect of Calcium Ion on Lipid Peroxide Formation in Endotoxemic Mice
Authors: Sakaguchi Shuhei , Ibata Hiroshi , Yokota Katsushi
Published: November 2013

The present study was conducted to determine the possible role of intracellular Ca2+ in lipid peroxide formation in endotoxin‐poisoned mice. Leakages of LDH isozyme and acid phosphatase in serum of mice fed a Ca2+‐deficient diet were remarkably increased after administration of 200 μg of endotoxin compared to that in endotoxin‐nontreated Ca2+‐deficient mice. Superoxide anion generation in liver of Ca2+‐deficient mice and in mice fed a normal diet greatly increased after endotoxin administration. On the contrary, after endotoxin injection there was scarcely any difference in SOD activity of liver of Ca2+‐deficient mice as compared to that in endotoxin‐nontreated Ca2+‐deficient mice. In spite of an increase of superoxide anion generation there was little or no effect of endotoxin administration on lipid peroxide formation in mice given a Ca2+‐deficient diet. In the mice treated with a Ca2+‐deficient diet, free radical scavenger levels (α‐tocopherol and nonprotein sulfhydryl) in liver tissue after endotoxin injection were markedly decreased compared to those in Ca2+‐deficient diet alone. Mice fed a normal diet exhibited a significant decrease of lipid peroxide level in liver by injection of endotoxin together with verapamil (10 mg/kg, s.c.). When mice fed a normal diet were injected with endotoxin, the state 3 respiratory activity showed a 49% decrease, and respiratory control ratio (RCR) of endotoxemic mice liver mitochondria was 38% lower than normal liver mitochondria. No difference could be observed in levels of state 3 and RCR between the mice given verapamil plus endotoxin and the normal mice. These findings suggest the possibility that Ca2+ may participate in the free radical formation in the liver during endotoxemia and also that Ca2+ may play an important role in the damage of liver mitochondrial function in endotoxemic mice.
Solubilization of IgG‐Binding Proteins from Group A and G Streptococci
Authors: Goward Christopher R. , Barstow David A.
Published: November 2013

The release of IgG‐binding proteins from the cell surface of streptococcal strains AR‐1 and G148 with various proteolytic enzymes, acid, alkali or SDS was investigated. The IgG‐binding proteins were purified by affinity chromatography using IgG‐Sepharose Fast Flow. After SDS‐polyacrylamide gel electrophoresis and immuno‐electroblotting the major proteins identified varied in relative molecular mass from 15,000 to 65,000 depending on the solubilizing agent used. The results showed that solubilization with trypsin gave the highest yield of IgG‐binding proteins, that strain G148 yielded about twice the amount of protein as strain AR‐1, and that elastase released an IgG‐binding protein of high relative molecular mass of 65,000.
A Non‐Radioactive DNA Probe for the Detection of Mycoplasma pulmonis in Murine Mycoplasmosis
Authors: Matsuzaki Michiya , Harasawa Ryô , Kimizuka Fusao , Koshimizu Kaoru
Published: November 2013

A non‐radioactive DNA probe for the detection of Mycoplasma pulmonis was developed by using photobiotin. The probe detected 0.3 ng chromosomal DNA of M. pulmonis. No cross‐hybridization was observed between the probe and the other murine mycoplasma species, M. arthritidis and M. neurolyticum.
A Protein Associated with Prodigiosin Formation in Serratia marcescens
Authors: Kobayashi Nobuharu , Ichikawa Yoichi
Published: November 2013

A protein associated to prodigiosin formation was found in Serratia marcescens. The protein was not found in nonpigmented strains and was correlated with the pigment level. The protein was about 100 kilodaltons (kDa) and was also found in nonpigmented bacteria of the pigmented strain grown in glucose medium, at high temperature, or under anaerobic condition. The 100 kDa protein was found not in the outer membrane and the periplasm, but in the inner membrane and/or the cytoplasm. The protein was also found singly or dominantly in pigment‐protein complexes and pigment‐localizing vesicles described in previous reports. These results suggest that the 100 kDa protein is associated with prodigiosin formation.
In vivo Influence of the Anti‐B220 Monoclonal Antibody Administration on the T Cell Differentiation in Mice
Authors: Assensi Victor , Himeno Kunisuke , Kawamura Ikuo , Sakumoto Misao , Nomoto Kikuo
Published: November 2013

B220, a pan‐B marker, is known to be also expressed on immature T cells of MRL/1pr or other congenic 1pr mice and minor population of immature thymocytes but not on peripheral T cells. In this study, we investigated in vivo the possibility whether B220 is one kind of premature T or prethymic T precursor cell marker as well as a pan‐B cell marker. A monoclonal antibody against B220 glycoprotein was intravenously injected every 2 days into various strains of mice. After the third administration of this antibody, thymocytes decreased remarkably compared with those from the rat IgG‐treated group, and spleen cells were also reduced significantly. Further, the number of cells expressing Thy‐1, Ly‐1, Lyt‐2, and asialo GM1 (asGM1) in the spleen were significantly reduced. On the contrary, the number of cells expressing surface IgM (sIgM) or B220 were increased by this treatment, especially after the 8th treatment. Some T‐dependent immunological functions including mitogenic responses to lectins and cytotoxic T cell activity were markedly suppressed but mixed lymphocyte reaction (MLR) and natural killer (NK) activity were rather augmented. Thus, B220 may be expressed on some kinds of T cell progenitor. Taken together, in vivo treatment with anti‐B220 antibody may influence differentiating stages of some T cells from bone marrow progenitors before or just after their homing into the thymus.
Protective Effect of N‐Acetyl Chitohexaose on Listeria monocytogenes Infection in Mice
Authors: Tokoro Akio , Kobayashi Makiko , Tatewaki Nobuhiko , Suzuki Ko , Okawa Yoshio , Mikami Takeshi , Suzuki Shigeo , Suzuki Masuko
Published: November 2013

A water‐soluble oligosaccharide, N‐acetyl chitohexaose (NACOS‐6) was able to enhance the protecting effect of BALB/c male mice against Listeria monocytogenes infection, when administered intraperitoneally 24 hr before the challenge with this microbe. Significant decrease in number of microbes within the peritoneal cavity, spleen, and liver from the mice of NACOS‐6‐administered group was not observed 1 day after the infection but 4 days after the infection. Administration of NACOS‐6 enhanced the delayed‐type hypersensitivity response against sheep red blood cells (SRBC) or heat‐killed L. monocytogenes. Splenic T lymphocytes from mice administered NACOS‐6 released macrophage activating factor (MAF). These results suggested that NACOS‐6 was also able to elevate the function of cellular immunity. Macrophages treated with a combination of NACOS‐6 and the culture supernatant of splenic T lymphocytes from mice administered NACOS‐6, “NACOS‐6 sup,” were found to exert a fairly strong growth‐inhibitory effect on L. monocytogenes. Interferon‐γ (IFN‐γ) and interleukin 2 (IL‐2) were able to enhance the growth‐inhibitory effect on L. monocytogenes by the NACOS‐6‐treated macrophages.
Binding of F41 and K99 Fimbriae of Enterotoxigenic Escherichia coli to Glycoproteins from Bovine and Porcine Colostrum
Authors: Lindahl Mats
Published: November 2013

F41 and K99 fimbriae of enterotoxigenic Escherichia coli were found to bind to periodate‐sensitive oligosaccharides of glycoproteins from bovine and porcine colostrum. Only a minor component of casein fractions (κ‐casein) possessed receptors for one type of fimbriae (K99). Both whey and fat globule membranes were rich in glycoproteins with receptor structures. Porcine colostrum seemed to contain a higher quantity of receptors than bovine colostrum.
Plasmid Profiles of Mycobacterium avium Complex Isolated from Swine
Authors: Masaki Shunichiro , Konishi Takao , Sugimori Giichi , Okamoto Atuko , Hayashi Yoshiyuki , Kuze Fumiyuki
Published: November 2013

Twenty strains of Mycobacterium avium complex (MAC) isolated from swine and five strains from humans were examined for drug susceptibility and plasmid content. Four strains of swine origin and two strains of human origin harbored plasmid DNAs differing in molecular weights. No relationship between plasmid contents and drug resistance was observed. Southern DNA‐DNA hybridization showed that small plasmids from swine MAC strains were homologous to those from human origin at the nucleotide level.
Protection of OK‐432, a Streptococcus pyogenes Preparation, against Lethal Infection of Mice with Herpes Simplex Virus
Authors: Harada Yusuke , Kurokawa Eri , Endo Hisao , Kimura Takao , Saito Motoo , Sugawara Yutaka , Irie Hiroshi , Ito Kyogo , Fujii Masahiko , Shimizu Yoshinobu , Kumagai Katsuo
Published: November 2013

We have studied the protective effect of OK‐432, a biological response modifier (BRM) of Streptococcus pyogenes origin, on the lethal infection of mice with herpes simplex virus (HSV)‐1. A single intraperitoneal (i.p.) injection of more than 10 μg of OK‐432, when given at least two days before the infection, gave a marked effect yielding nearly 100% protection against ordinarily lethal infection. The protection was independent of the amount of infected virus inoculated. When given after the infection, the agent even at the maximal dose (100 μg), produced only a marginal effect. A single i.p. administration of OK‐432 augmented the natural killer (NK) activity of peritoneal exudate cells and spleen mononuclear cells in mice 2 to 3 days after injection of OK‐432, coinciding with the times when it induced a survival effect on HSV‐infection. Treating OK‐432‐treated mice with a combination of an anti‐macrophage agent, silica, and an anti‐NK cell agent, anti‐asialo GM1 serum, before infection diminished the antiviral effect of OK‐432. The OK‐432 protection against HSV infection was also markedly diminished in athymic nude mice. Thus, the protective effect of OK‐432 on lethal HSV infection seems to be based on the activation of NK cells, macrophages, and T lymphocytes.
Specific Immunoglobulin Response in Mice Immunized with Porins and Challenged with Salmonella typhi
Authors: Sharma Praveen , Ganguly Nirmal Kumar , Sharma Bal Krishan , Sharma Saroj , Sehgal Rakesh
Published: November 2013

Specific antiporin antibody (IgG, IgM, and IgA) response was studied in control, infected, immunized‐infected, and immunized mice. The activity of specific IgG immunoglobulins was found to be the highest in immunized and immunized‐infected groups in which 87.5% protection had been observed by laboratory potency test in mice; the next‐highest activities were those of IgM and IgA immunoglobulins. However, in the infected group the activity of specific IgM and IgG immunoglobulins was almost similar.
Characterization and Deposition of the Proteins in the Outermost Layer of Bacillus megaterium Spore
Authors: Takubo Yoshihiro , Okuda Masayuki , Takemura Isao , Haruna Fumiyo , Sawatari Akiko , Nishihara Tsutomu , Kondo Masaomi
Published: November 2013

It was proved that three spore coat proteins of 48, 36, and 22 kDa (P48, P36, and P22) were the components of the outermost layer (OL) of Bacillus megaterium ATCC 12872 spore by analysis of the isolated OL. And it was indicated that these proteins were deposited not by disulfide bond, but by ionic and/or hydrophobic bonds on the spore. Among them, P36 and P22 were expected to be located on the very surface of the spore by immunological analysis. In the OL deficient mutant of B. megaterium ATCC 12872, MAE05, whose spore was lacking in these OL proteins and galactosamine‐6‐phosphate polymer, both P36 and P22 were present in the mother cell cytoplasm and deposited on the forespores, but they disappeared with the lysis of mother cells. An OL protein‐releasing factor having proteolytic activity was detected in the culture supernatant at the late sporulating stage of both the wild‐type and the mutant strains. But the factor could not act on the proteins of the mature spores and the forespores at t10 (tn indicates n hr after the end of exponential growth) of the wild‐type strain. Moreover, P36 and P22 were found in the spores of a revertant of MAE05 which could form galactosamine‐6‐phosphate polymer, suggesting that this sugar polymer played the role in protecting the OL proteins against the protease‐like substance after the deposition.
Isolation and Characterization of Cytoplasmic Fibrils from Treponemes
Authors: Masuda Kuniyoshi , Kawata Tomio
Published: November 2013

Electron microscopy of Triton X‐100‐treated whole cells of an oral treponeme, Treponema sp. strain E‐21, revealed that six cytoplasmic fibrils (CFs) helically wound as a bundle in the cytoplasm. The CFs were isolated and purified by disruption and solubilization of the cells followed by CsCl density gradient centrifugation. The purified CF preparation contained mostly fibrils of about 9 nm in width and very small amounts of thinner strands of about 3 nm in diameter. The CFs were apparently seen to be a tubular structure, but the isolated CFs had narrowed sites of about 4–5 nm in width lacking lumen‐like images, possibly representing twisted sites. Thus, the CF did not seem to be a tubular structure. The purified CFs were composed of one major 82 kDa protein and a few minor proteins. The CFs were destructed by treatment with proteases, 8 m urea or 4 m guanidine hydrochloride. Very low tyrosine content (0.76 mol %) and lack of methionine were characteristic features for the 82 kDa protein. The CF preparations from the other five treponemes including Treponema phagedenis and T. denticola also had 82 kDa proteins as a major component, and the 82 kDa proteins of all of the treponemes had a common antigen when examined by using antiserum against the 82 kDa protein from Treponema sp. strain E‐21. Furthermore, the 82 kDa protein was demonstrated to be a principal component of the CFs of all the treponemes by immunoelectron microscopy.
Sugar Composition of the Polysaccharide Portion of Lipopolysaccharides Isolated from Non‐O1 Vibrio cholerae O2 to O41, O44, and O68
Authors: Kondo Seiichi , Hisatsune Kazuhito
Published: November 2013

The sugar composition of the polysaccharide portion of lipopolysaccharides (LPS) was determined for 42 serovars of non‐O1 Vibrio cholerae, i.e., from serogroups O2 to O41, O44, and O68. On the basis of their compositional sugar pattern, they were classified into 24 chemotypes. 2‐Keto‐3‐deoxyoctonate (KDO) was totally undetectable by the conventional color test (Weissbach's reaction) under the conventional hydrolysis conditions. Instead, a kind of KDO‐like substance, which was positive in the reaction but not identical to KDO, was found in serogroup O19. Fructose, a characteristic sugar constituent of O1 V. cholerae LPS, was found in 33 serogroups but was absent from nine serogroups, approximately 20% of the members of this group so far examined.
Acetylcholine Inhibition of Escherichia coli Chemotaxis for Aspartate
Authors: Shinozawa Takao , Fukunaga Shinya
Published: November 2013

Though acetylcholin per se was not attractant or repellent for Escherichia coli, it was found that acetylcholine inhibits the chemotaxis of the bacteria for aspartate. The inhibition appeared at 10−5 m of acetylcholine and at 10−2 m the inhibition reached 50%.
Demonstration of a Surface Antigen of Clostridium tyrobutyricum by Use of Immunoblotting with a Monoclonal Antibody
Authors: Gueguen Françoise , Robreau Georges , Talbot Françoise , Malcoste Roger
Published: November 2013

A monoclonal antibody, prepared against whole cells of Clostridium tyrobutyricum, recognized a surface antigen extracted by heat treatment or by hot phenol‐water treatment. This antigen, after analysis by polyacrylamide gel electrophoresis and immunoblotting, has been shown to present a regularly‐spaced ladder pattern similar to those shown by the lipopolysaccharide of many gram‐negative bacteria. The proteinase K has been shown to have no effect on the recognition of this epitope by the monoclonal antibody. On the contrary, the inhibition of the antigen reactivity to the monoclonal antibody after a mild periodate oxidation suggests the involvement of a carbohydrate moiety in the epitope. Moreover, the SDS‐PAGE analysis of phenol‐water extracts has shown an additional compound, detected by silver staining but not recognized by the monoclonal antibody.
Cell Surface of a Tetrads‐Forming Mutant of Micrococcus luteus: Chemical Treatment of the Cells and Teichuronic Acids on the Surface
Authors: Monodane Toshihiko , Tokunaga Michiko , Torii Mitsuo
Published: November 2013

Tetrads‐forming mutant MT cells of Micrococcus luteus, both treated with chemical reagents and non‐treated, were observed with a scanning electron microscope (SEM). The agglutinability of the cells with antiserum containing anti‐teichuronic acid antibody was examined. The binding of protein A‐gold particles to the cells, mediated with the antiserum, was also observed with SEM. A tetrad surface, not surface of each of four “unit monococci” constituting a tetrad, consisted of two or three smooth areas with borders. The difference in the surface features between M. luteus wild‐type IFO 3333 (Monodane et al, Microbiol. Immunol. 33: 165–174, 1989) and the mutant MT cells is discussed.
Isolation and Molecular Characterization of a Serotype 9 Human Rotavirus Strain
Authors: Nakagomi Toyoko , Ohshima Atsushi , Akatani Kaoru , Ikegami Nobuko , Katsushima Noriko , Nakagomi Osamu
Published: November 2013

A human rotavirus strain, designated AU32, that belongs to serotype 9 was isolated and was compared by RNA‐RNA hybridization with recently established two serotype 9 strains (WI61 and F45) as well as other prototype human strains. These three strains exhibited a very high degree of homology with one another and shared a high degree of homology with strains belonging to the Wa genogroup but not with strains belonging to either the DS‐1 or AU‐1 genogroup. These results suggest that genetic constellation of the serotype 9 strains is similar to that of the commonest human rotavirus despite the recent recognition of this serotype.
Arrest of DNA Replication of Macrophages in BCG Granuloma and Peritoneal Exudates by Bacteria
Authors: Kutsukake Hiroshi , Nagao Shigeki , Tanaka Atsushi
Published: November 2013

Previously we found that bacterial surface substances induced the suppression of DNA synthesis of macrophages. We examined in the present study whether DNA synthesis of macrophages could be similarly suppressed by whole bacteria themselves. For this purpose we isolated macrophages from BCG granuloma of guinea pigs and rats. The macrophages from both of the animals gave essentially the same results. No isolated macrophages containing bacilli were found to incorporate 3H‐thymidine when tested by autoradiography. Further, DNA replication of peritoneal exudate macrophages was markedly and rapidly suppressed in vitro upon phagocytosis of various kinds of bacteria but not of non‐bacterial preparations. A close correlation was found between granuloma formation and inhibition of 3H‐thymidine incorporation by MDP, its analogs, and various bacteria. These findings suggest that macrophages suppress their DNA replication when they phagocytose bacteria and that they can discriminate between bacteria and non‐bacterial preparations.
Enhancement of Cytotoxicity of Active Macrophages by Mycoplasma: Role of Mycoplasma‐Associated Induction of Tumor Necrosis Factor‐α (TNF‐α) in Macrophages
Authors: Arai Sumio , Furukawa Mayumi , Munakata Tetsuo , Kuwano Koichi , Inoue Hiroyuki , Miyazaki Teinosuke
Published: November 2013

Tumor necrosis factor‐alpha (TNF‐α) ‐inducing activity of several mycoplasmas including Mycoplasma pneumoniae, a causative agent in human respiratory infectious diseases, was investigated. Purified peritoneal macrophages from BALB/c mice markedly enhanced their cytotoxic activity to Meth A cells, when cultured with either viable or non‐viable mycoplasmas. The supernatants of the macrophage culture with mycoplasmas, M. pneumoniae and Acholeplasma laidlawii, showed the potent cytotoxic activity to TNF‐α‐sensitive L cells but not to TNF‐α‐insensitive L cells. Addition of anti‐TNF‐α antiserum inhibited completely the cytotoxic activity of these supernatants, indicating that a major part of the cytotoxic activity might be due to TNF‐α. Various other mycoplasmas, either glucose‐ or arginine‐utilizing species, as far as tested showed also the potent activity to produce TNF‐α. These results strongly suggest the possibility that mycoplasmas possess the activity of TNF‐α induction which might be responsible for a part of enhancement of cytotoxic activity of macrophages and resistance to infection with mycoplasmas in vivo.
Characteristics of Immunosuppressive Macrophages Induced in Host Spleen Cells by Mycobacterium avium Complex and Mycobacterium tuberculosis Infections in Mice
Authors: Tomioka Haruaki , Saito Hajime , Sato Katsumasa
Published: November 2013

The profile of generation and characteristics of immunosuppressive macrophages (Mφs). which suppress the ConA‐mitogenic response of spleen cells (SPCs), in host CBA/JN mice during the course of Mycobacterium avium complex (MAC) and M. tuberculosis (MT) infections were investigated. In both infections, a marked reduction in ConA mitogenic response of splenic T cells was seen around 2 weeks after infection, and this was accompanied by generation of potent immunosuppressive Mφs in the SPCs of infected mice. The suppressive activity was much stronger in MT‐infected mice than in MAC‐infected ones. In both infections, the large part of the suppressive Mφs exhibited suppressor activity that depended on the arachidonic acid cascade, particularly mediated by prostaglandins (PGs), and the remainder showed the suppressor action independent from PGs. The unique finding of this study is that the generation of IL‐2 reactive T cell populations in SPCs in response to ConA signal was markedly inhibited by the MAC‐ and MT‐induced immunosuppressive Mφs, whereas the suppressive Mφs failed to reduce the IL‐2‐producing ability of splenic T cells. In any case, the present results indicate a close similarity in immunosuppressive Mφs induced by MAC and MT infections.