bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunotyping of Chlamydia psittaci by Indirect Immunofluorescence Antibody Test with Monoclonal Antibodies
Authors: Takahashi Tatsufumi , Takashima Ikuo , Hashimoto Nobuo
Published: November 2013

Monoclonal antibodies against pigeon and budgerigar strains of Chlamydia psittaci were used to classify the immunotypes of C. psittaci strains by an indirect immunofluorescence antibody (IFA) test. Thirty‐three C. psittaci strains from pigeons and 24 from budgerigars were divided into three immunotypes (P‐I, P‐II, and P‐III) and (B‐I, B‐II, and B‐III), respectively. Two strains from human psittacosis patients were identified as P‐III and B‐I, coinciding with the epidemiological evidence of each human infection. Two strains from psittacine birds, a parrot and a parakeet, were identical to the B‐II immunotype. Other mammalian strains were quite distinct from avian strains in their IFA reaction with the monoclonal antibodies.
Role of a Guanine Nucleotide‐Binding Regulatory Protein in the Hydrolysis of Phosphatidylinositol 4,5‐Bisphosphate in a Human T Cell Line
Authors: Hasegawa‐Sasaki Hiroko , Lutz Frieder , Sasaki Terukatsu
Published: November 2013

The CD3(T3)/antigen receptor complex appears to function by transducing an antigen signal presented by macrophages into the hydrolysis of phosphatidylinositol 4,5‐bisphosphate [PtdIns(4,5)P2]. In order to find out how the CD3/antigen receptor complex regulates the hydrolysis of PtdIns(4,5)P2 to diacylglycerol and inositol trisphosphate, we investigated the possible role played by a guanine nucleotide‐binding regulatory protein in PtdIns(4,5)P2 hydrolysis in a human T cell leukemia line, JURKAT. JURKAT cells were made permeable to Al3+, F−, GTP, and a nonhydrolyzable GTP analogue, guanosine 5′‐O‐(3‐thiotriphosphate) (GTPγS), by treatment with pseudomonal cytotoxin. In the presence of AlCl3 NaF stimulated the release of inositol phosphates in the cytotoxin‐treated JURKAT cells. NaF plus AlCl3 induced increases in inositol tris‐, bis‐, and mono‐phosphates and decreases in PtdIns(4,5)P2, phosphatidylinositol 4‐phosphate, and phosphatidylinositol within 5 min after addition to the cytotoxin‐treated cells at 37 C. GTPγS stimulated, to some extent, polyphosphoinositide hydrolysis in the cytotoxin‐treated JURKAT. The cytotoxin‐treated JURKAT cells retained the ability to respond to anti‐Leu‐4 with polyphosphoinositide hydrolysis. It has been shown that Al3+ in the presence of F− modulates the activity of various guanine nucleotide‐binding regulatory proteins. Therefore, the results obtained in this study indicate that a guanine nucleotide‐binding regulatory protein regulates the polyphosphoinositide breakdown in JURKAT cells by influencing phosphodiesterase activity.
Characterization of Mouse Monoclonal Antibodies to Human Interferon‐Gamma
Authors: Yamamoto Yoko , Miyata Keizo , Ueda Masamichi , Kawade Yoshimi , Matsumoto Kayoko , Tsukui Kazuo
Published: November 2013

Mouse monoclonal antibodies (mAb) to human interferon‐gamma (HuIFN‐γ) were characterized. The mAbs studied—E4‐18, G4‐15, and SAT‐1—which are all IgGl‐type, reacted to all HuIFN‐γ molecular species, both glycosylated and non‐glycosylated. Affinity constants calculated of E4‐18 and G4‐15 didn't have considerable differences for both kinds of HuIFN‐γ (1–3 × 108 liter/mol), but SAT‐1 had a difference—a higher value (1010 liter/mol) for the former than for the latter (8 × 108 liter/mol). In epitope specificity, the results suggested that E4‐18 and G4‐15 recognized an overlapped region remote from the region of SAT‐1. Competition experiment using synthetic peptides suggested that epitope of G4‐15 is around N9‐26 of the HuIFN‐γ sequence. Those mAbs could be used for sandwich radioimmunoassay of HuIFN‐γ using double mAbs in two combinations, one (G4‐15/E4‐18) based on dimer forms of HuIFN‐γ and the other (SAT‐1/E4‐18) based on epitope difference. The mAbs are all neutralizing antibodies in which SAT‐1 neutralized at a lower concentration than did G4‐15, and at a much lower one than did E4‐18. The receptor binding of HuIFN‐γ was inhibited by mAbs G4‐15 and SAT‐1. Efficacy of G4‐15 and SAT‐1 for the inhibition correspond with that for neutralization.
Enhanced Neutralization of Flavivirus by Monoclonal Antibodies against Negishi Virus
Authors: Motohide Yukage , Takashima Ikuo , Hashimoto Nobuo
Published: November 2013

Ten monoclonal antibodies against Negishi virus were classified into 3 groups and 7 types according to the reaction patterns to Negishi virus by the hemagglutination inhibition (HI) test and by several kinds of neutralization tests. When kinetic neutralization was applied to these monoclonal antibodies at the same HI titer of 1:64, the initial slope and the persistent fraction of the kinetic curve was varied in each antibody. In the double‐kinetic neutralization test, neutralization did not proceed further when the second antibody was the same type as the first one. When the second antibody was a different type from that of the first one, neutralization proceeded further. The mixtures of 4 monoclonal antibodies classified as different groups and types remarkably enhanced neutralization in the kinetic assay. These results suggested that an assortment of antibodies is needed for effective neutralization of Negishi virus and that a multi‐hit model is likely operating in the neutralization of Negishi virus.
Replication of Human Immunodeficiency Virus in Two T‐Cell Lines and Their Application as Antigens for Immunofluorescence
Authors: Kawatani Toshio , Hattori Naohiko , Kurimura Takashi
Published: November 2013

Two T‐cell lines, TALL‐1 and CCRF‐CEM, were infected with human immunodeficiency virus (HIV), strain LAV, to explore the time course of the appearance of various virus specific antigens, and to establish an antibody assay system by indirect immunofluorescence (IF). These cells were infected with LAV at two different input multiplicity of infection (MOI). Antigens were tested by Western blot analysis (WB) and IF. Antigens for WB were extracted from the infected cells at various times after infection, but pooled sera of American HIV carriers could not recognize gp41 or gp160. Antigen expression was highest in CCRF‐CEM, but, as the antigen for IF, TALL‐1 infected at the MOI of 8.0 was the most suitable 7 days after infection, because it includes a fairly large number of uninfected cells, which served as the internal control.
Immunological Detection of 22K Protein in Sporulating Cells of Bacillus megaterium ATCC 12872
Authors: Nishihara Tsutomu , Sakagami Kimie , Tani Katsuji , Imagawa Masayoshi , Kondo Masaomi
Published: November 2013

The synthesis and deposition of 22,000‐dalton (22K) spore coat protein were examined immunochemically on the sporulating cells of Bacillus megaterium ATCC 12872 using the antibody to purified 22K spore coat protein. This antibody cross‐reacted with 44K and 25K proteins in immunoblot analysis of dormant spore coat proteins. Immunoblot analysis on the sporulating cells showed that 22K protein was detected from t8 in forespore coat protein fractions. Sandwich enzyme immunoassay revealed that 22K protein in the spore coat protein fraction appeared at t6 and reached a plateau at t9, and 22K protein in the mother cell cytoplasmic fraction was detected at only t7 and t8 at a very low level.
Upstream Region of Hepatitis B Virus S Gene Responsible for Transcriptional Stimulation by Dexamethasone
Authors: Masuda Michiaki , Lee Ganhon , Yuasa Tazuko , Yoshikura Hiroshi
Published: November 2013

Transcriptional regulation of hepatitis B virus (HBV) surface antigen (HBs Ag) gene was studied in human hepatoma‐derived cell lines. Treatment with dexamethasone (Dex; 1 μM) induced an increase in the smaller HBs‐mRNA initiated within Pre‐S region encoding S and Pre‐S2 proteins, but not the larger HBs‐mRNA initiated in the further upstream encoding Pre‐S1 protein. The Bg1II‐MstII fragment (map position 2425–3201) in the upstream of the S gene was used as a transcriptional promoter of chloramphenicol acetyltransferase (CAT) gene. The CAT activity brought about by this construct in the transient assay was elevated by 5‐fold in the presence of Dex. Deletion analysis localized the sequence required for the full response to Dex within a 590‐base pair fragment in the upstream of the transcriptional initiation site of the smaller HBs‐mRNA. And this fragment contained the binding site for the nuclear factor I (NF‐I), which might have some role in Dex‐dependent transcriptional stimulation.
Interaction of Herpes Simplex Virus Type 2 with a Rat Glioma Cell Line
Authors: Sakihama Kuniharu , Eizuru Yoshito , Minamishima Yoichi
Published: November 2013

The interaction between herpes simplex virus type 1 (HSV‐1) and type 2 (HSV‐2) and two neural cell lines, mouse neuroblastoma (N1E‐115) and rat glioma (C6‐BU‐1), was investigated. N1E‐115 cells were permissive to both types of HSV. In C6‐BU‐1 cells, on the other hand, all the HSV‐1 strains tested so far showed persistent infection, and the infectious virus of HSV‐2 strains disappeared spontaneously. The HSV‐2‐infected C6‐BU‐1 cells were positive for HSV‐2‐specific DNA sequences, virus‐specific RNA, HSV‐2‐specific antigens and thymidine kinase activity, when no infectious virus was detected. The HSV‐2 was reactivated from those C6‐BU‐1 cells by superinfection with murine cytomegalovirus (MCMV), but not with UV‐irradiated MCMV or human cytomegalovirus. The reactivated HSV‐2 was identical to the parental virus, when examined by restriction endonuclease cleavage analysis.
Sporadic Carriers of Human T‐Lymphotropic Virus Type I in Northern Egypt
Authors: El‐Farrash Mohamed A. , Badr Mohamed F. , Hawas Samia A. , El‐Nashar Nariman M. , Imai Joko , Komoda Haruko , Hinuma Yorio
Published: November 2013

Sera from 3,158 individuals living in northern Egypt were tested for the presence of antibodies against human T‐lymphotropic virus type I (HTLV‐I) by the newly developed particle agglutination (PA) test. Ten sera gave a positive reaction in the PA test. Eight of these sera were examined further by Western blotting and all of them gave several bands corresponding to HTLV‐I structural proteins. Two of the 8 sera gave positive results in the indirect immunofluorescence test. The results indicate the presence of HTLV‐I carriers in this area, although at very low incidence (0.063%).
Characteristics of a Colicin Plasmid in Escherichia coli Strain B177 Isolated from a Septicemic Calf
Authors: Harasawa Ryô , Miura Hiroyuki , Kaneuchi Choji
Published: November 2013

A colicin plasmid in Escherichia coli strain B177 isolated from a septicemic calf was characterized. The colicin type was identified as ColV by using reference ColV producers. The colicin plasmid was labeled with transposon Tn903 and subjected to conjugation. The transconjugants examined suggest that the colicin plasmid confers serum resistance. There was no difference in siderophore utilization ability between the transconjugants and host strain SF800. Bioassay for siderophore suggests that the colicin plasmid specifies the production of iron‐chelating compounds available for the host strain.
Restriction Endonuclease DNA Analysis of Antigenic Variants of Leptospires Selected by Monoclonal Antibodies
Authors: Yamaguchi Tsuyoshi , Ono Etsuro , Ito Toshihiro , Yanagawa Ryo
Published: November 2013

The genome of antigenic variant CV(CT3)‐1 derived from Leptospira interrogans serovar canicola was compared by cleavage with restriction endonucleases with the parent and serovar bafani, to which the variant was serologically most closely related. No differences were observed between the parent and variant in DNA restriction endonuclease patterns using eight restriction endonucleases. Serovar bafani was different in the patterns from the parent and antigenic variant CV(CT3)‐1. The two antigenic variants derived from serovar hebdomadis, HV(H16)‐1 and HV(H19)‐1 which belonged serologically to serovars jules and hebdomadis, respectively, were compared by restriction endonuclease DNA analysis with the parent and serovar jules. No differences were observed between the parent and variants in DNA restriction endonuclease patterns using the same enzymes. But some differences were observed in DNA restriction endonuclease patterns between HV(H16)‐1 and serovar jules. Thus, the antigenic variant selected from the parent by the anti‐parent monoclonal antibody and serologically different from the parent, being identified either as a new serovar or as a known one, was found to be similar to the parent by the restriction endonuclease DNA analysis.
Detection and Specificity of a New Antigen in Candida tropicalis and Its Evaluation by Taxonomic DNA Analyses
Authors: Hamajima Kenji , Nishikawa Akemi , Shinoda Takako , Fukazawa Yoshimura
Published: November 2013

Monospecific factor serum for identifying Candida tropicalis was obtained either from rabbit antiserum to heated cells of C. tropicalis M 1519 (S 96) or from antiserum to C. tropicalis IFO 1400, by adsorption with heated cells of Candida albicans serotype A, or C. albicans (A) and Candida krusei, respectively. We designated this adsorbed serum factor t serum. The monospecific factor serum reacted with 31 out of 32 strains of C. tropicalis, only when tested on heat‐treated cell antigens, whereas it did not react with any of 72 strains of the six other medically important species of Candida. The morphological and physiological characteristics of the one strain of C. tropicalis that did not react with the factor t serum, designated the t−‐strain, were shown to be similar to those of the type strain of C. tropicalis by most of the methods employed for identifying Candida. Therefore, cell wall mannan from the t−‐strain was compared with that from several typical strains of C. tropicalis for its specificity by the precipitation reaction and also for its 1H‐nuclear magnetic resonance spectrum. The results showed that these mannans are similar to each other serologically and physicochemically, suggesting that the new antigen t is not mannan. Taxonomic characterization of the t−‐ and several typical strains of C. tropicalis was carried out by determining the mol% G + C of their DNA and also their DNA homology. Although the mol% G + C values of four typical strains of C. tropicalis were fairly similar (35.2 to 36.2 mol% by the Tm method and 35.5 to 36.4 mol% by the HPLC method), the t−‐strain had a G + C content of 44.1 (Tm) and 43.3 (HPLC) mol%. Furthermore, the DNAs of the t−‐strain and the type strain of C. tropicalis showed only 18.2% relatedness. These results suggest that the antigen corresponding to serum factor t exists only in the cell wall of C. tropicalis strains, not in those of the other medically important Candida, and that the t−‐strain should not be classified as C. tropicalis. In conclusion, the taxonomic value and usefulness of factor t serum is primarily for differentiating C. tropicalis from C. albicans serotype A serologically.
Method of Genomic DNA Cloning by the Combination of Cosmid Shuttle Vector and Monoclonal Antibody
Authors: Tagawa Masatoshi , Ito Toshihiro , Kanno Masamoto , Okitsu Akira , Sakamoto Tohru , Imai Kenji , Kuwabara Ichiro , Koseki Haruhiko , Matsubara Hisahiro , Lau Yun‐fai , Taniguchi Masaru
Published: November 2013

We report here the strategy to isolate the DNA fragment of any species origin which encodes cell surface antigen by using cosmid library transfection and cell sortings with a monoclonal antibody. We took the mouse melanoma antigen defined by monoclonal antibody as a model system and rescued the genomic DNA by in vitro packaging, showing the feasibility of this procedure.
Relationship between Susceptibility and Immune Response to Staphylococcal Exfoliative Toxin A in Mammalian Species
Authors: Machida Katsuhiko , Sakurai Susumu , Kondo Isamu , Ikawa Satio
Published: November 2013

Staphylococcal exfoliative toxin A (ETA) had a splitting effect at the granular layer of skin in humans and neonatal mice, but not in rabbits, guinea pigs, golden hamsters, or rats. Besides its splitting effect, ETA could stimulate productions of neutralizing antibody to ETA in rabbits, rats and B10D2 mice, but not in golden hamsters, guinea pigs, or ICR, HRS/J, and C57BL/10 mice. In our epidemiological investigation of human sera, the percentage of antibody to ETA in sera obtained from patients with impetigo (8%) was lower than those in sera of healthy males (23%) and females (29%). The relationship between susceptibility and immune response to ETA in these mammalians could be divided into three groups: the possession of resistant skin and high production of antibody to ETA; the possession of resistant skin and low production of antibody to ETA; the possession of sensitive skin and various titers of antibody to ETA.
The Caries Inhibitory Effects of GOS‐Sugar in vitro and in Rat Experiments
Authors: Ooshima Takashi , Fujiwara Taku , Takei Tsutomu , Izumitani Akira , Sobue Shizuo , Hamada Shigeyuki
Published: November 2013

The caries inhibitory activity of GOS‐sugar (panose‐ and maltose‐rich sugar mixture) was examined and compared with that of sucrose, maltose, or glucose in in vitro and in vivo experiments. Streptococcus mutans MT8148R (serotype c) and Streptococcus sobrinus 6715 (g) did ferment GOS‐sugar and produce acid in a similar way as with maltose and glucose. However, GOS‐sugar could not be a substrate for the glucosyltransferases (GTases) of these mutans streptococci to synthesize the water‐insoluble glucan. Also, it significantly inhibited not only the synthesis of water‐insoluble glucan from sucrose by the crude GTases but also the sucrose‐dependent adherence of these cells to a glass surface. In particular, adherence of growing cells of 6715 was markedly inhibited by the presence of GOS‐sugar. GOS‐sugar was found to induce significant but minimal dental caries in SPF rats infected with either MT8148R or 6715. Furthermore, the replacement of half of the dietary sucrose content with GOS‐sugar resulted in a significant reduction of caries development in rats infected with strain 6715.
Genome Typing of Adenovirus Strains Isolated from Conjunctivitis in Japan, Australia, and the Philippines
Authors: Guo Deng Fu , Shinagawa Morikazu , Aoki Koki , Sawada Harumi , Itakura Shim , Sato Gihei
Published: November 2013

DNA restriction endonuclease analysis for genome typing of adenovirus (Ad) DNA was carried out on a total of 65 Ad isolates including serotypes Ad4, Ad7, Ad8, Ad11, Ad19, and Ad37 from patients with epidemic keratoconjunctivitis and acute conjunctivitis obtained in Japan from 1982 to 1986, Australia from 1973 to 1986, and the Philippines in 1984. All 4 isolates of Ad7 in Australia were Ad7b. Four of 6 Ad11 isolates obtained in Japan were typed as Ad11 prototype (Ad11p), and the remaining were identified to be new genome types, designated tentatively as Ad11c and Ad11d. An isolate of Ad11 obtained in Australia was typed as Ad11c. Nine Ad8 isolates in Australia and in the Philippines were typed as Ad8p, but 11 Ad8 isolates in Japan were Ad8b. Thirteen Ad19 isolates were identified as Ad19a. All 3 isolates of Ad37 in Japan and three isolates in Australia before 1982 were typed as Ad37p, however, 5 isolates in Australia after 1983 were identified as a new genome type, designated as Ad37d. In Japan, 10 isolates of Ad4 were identified as Ad4a.
Ultrastructural Study on Japanese Isolates of Spotted Fever Group Rickettsiae
Authors: Uchiyama Tsuneo , Uchida Takahiro
Published: November 2013

Japanese isolates of spotted fever group rickettsiae were observed under a transmission electron microscope. In Vero cells persistently infected with Japanese isolates, small numbers of intracytoplasmic rickettsiae were seen. On the other hand, moderate numbers of rickettsiae were found in the cytoplasm of productively infected BHK cells. The electron‐lucent, halo‐like zone was found to surround organisms in the cytoplasm of their host cells, which is a prominent characteristic of spotted fever group rickettsiae. Fine structural features of the cell wall revealed thin outer and thick inner leaflets like those observed in other spotted fever group rickettsiae.
Depression of T Cell‐Mediated Immunity and Enhancement of Autoantibody Production by Natural Infection with Microorganisms in Spontaneously Hypertensive Rats (SHR)
Authors: Takeichi Noritoshi , Hamada Jun‐ichi , Takimoto Masato , Fujiwara Kosaku , Kobayashi Hiroshi
Published: November 2013

We studied the effects of breeding conditions on the development of immunological abnormalities in spontaneously hypertensive rats (SHR) with congenital T cell depression. The depression of T cell functions, the production of natural thymocytotoxic autoantibody (NTA), and the development of polyarteritis nodosa were more evident in SHR reared under a conventional (CV) environment than in specific‐pathogen‐free (SPF) SHR bred in a semi‐barrier system. Enhancement of these immunologic abnormalities was also observed by the conventionalization of SPF‐SHR. A high frequency of antibodies to mouse hepatitis virus (MHV), Sendai virus, and Mycoplasma pulmonis was detected in CV rat sera, whereas no antibodies were detected in SPF‐SHR. The experimental infection of Sendai virus induced the enhancement of T cell depression and of NTA production in SPF‐SHR. We interpret these results to mean that the natural infection of microorganisms causes an acceleration of immunologic abnormalities in SHR reared in a CV environment.
Protection of Mice against Bacterial Infection by Oral Administration of Bacterial Lipopolysaccharide
Authors: Fujiki Tetsuro , Kutsukake Hiroshi , Imai Katsuyuki , Tanaka Atsushi
Published: November 2013

The effect of orally administered bacterial lipopolysaccharide (LPS) on host resistance against bacterial infections was studied. LPS orally given for 5 consecutive days prior to infection caused no apparent toxic effect and protected mice against Pseudomonas aeruginosa and Listeria monocytogenes infections.
Seroepidemiology of Herpes Simplex Virus in Yogyakarta, Indonesia
Authors: Haksohusodo Suwardji , Sakuma Shunji , Kino Yoichiro , Mori Ryoichi
Published: November 2013

Sera from 487 individuals in Yogyakarta, Indonesia, were tested for antibodies to herpes simplex virus (HSV) by a passive hemagglutination method. Age‐specific incidence rates for antibodies to HSV were calculated. For sera from persons other than prostitutes, in the age group from 10 to 19, the positive rate was 48% but in the age group higher than 20, it was more than 87%. Fifty of 59 pregnant women (85%) were positive. The positive rate and the distribution of antibody levels in prostitutes were higher than in the general population.
The Effect of GD3 Ganglioside Obtained from Bovine Lymphosarcoma on Bovine Normal Mononuclear Cell
Authors: Yasutomi Yasuhiro , Onuma Misao , Watarai Shinobu , Yasuda Tatsuji , Kirisawa Rikio , Kawakami Yoshimi
Published: November 2013

The effect of immunological function of GD3 on normal bovine lymphocyte was examined with various in vitro assay systems. The GD3 level in sera from enzootic bovine leukemia (EBL) cattle was significantly increased compared with that of normal cattle (EBL: 0.62±0.24 μg/ml; normal cattle: 0.33±0.09 μg/ml, P<0.05). Lymphocyte blastogenesis elicited by concanavalin A was inhibited by addition of a 50 μg/ml concentration, or more, of GD3. Inhibitory effect of GD3 in IL‐2‐dependent T cell line and EBL tumor cell line was hardly observed compared with normal peripheral blood mononuclear cells. GD3 also inhibited mixed lymphocyte reaction and allo cytotoxic T lymphocyte reaction.
Genomic Difference of Herpesvirus of Turkeys at Low and High Passage Levels in Culture of O1 and FC126 Strains
Authors: Hirai Kanji , Ikuta Kazuyoshi , Mikami Takeshi , Kato Shiro
Published: November 2013

Serial passages in culture of herpesvirus of turkeys resulted in structural genomic changes at the regions common to two virus strains with loss of their protective ability against Marek's disease.
Characterization of Putrescine Production in Nongrowing Vibrio parahaemolyticus Cells in Response to External Osmolality
Authors: Yamamoto Shigeo , Yamasaki Keiko , Takashina Kimiko , Katsu Takashi , Shinoda Sumio
Published: November 2013

Nongrowing Vibrio parahaemolyticus cells rapidly produced putrescine (Put) from added arginine when subjected to a low osmotic stress. This phenomenon was characterized in connection with a regulatory mechanism of the responsible enzymes, arginine decarboxylase (ADC) and agmatine ureohydrolase (AUH). NaCl, KCl, LiCl, sucrose, and glycerol were used as solutes to prepare the resuspending media with various osmolalities. Regardless of whether the solutes were electrolytes or non‐electrolytes, exposure of cells to low osmolality brought about instantaneous increases in both intra‐ and extracellular Put contents without significant changes in the contents of other polyamines. This acceleration in Put production was accompanied by no increases in the specific activities of ADC and AUH. On the other hand, when cells were exposed to the osmolality equivalent to 2 or 5% NaCl, all solutes except for glycerol did not cause a remarkable variation in the intracellular Put content, while the amount of Put in the medium varied depending on the solute used; sucrose and glycerol still greatly prompted Put production, as judged by high Put contents in the media, even at the osmolality equivalent to 5% NaCl. The cation efflux from cells, measured as the K+ release, was observed whenever the increase in Put production occurred. Furthermore, in vitro experiments showed that NaCl and KCl inhibited ADC to a similar extent, about 70% inhibition being observed at 200 mm. However, AUH was not affected by these compounds. These results suggest that the reduction in the concentrations of Na+ and K+ predominantly present in cells may cause the increase in activity of the preexisting ADC, which leads to the enhancement of Put production.
Subcellular Distribution of the Transmissible Agent in Creutzfeldt‐Jakob Disease Mouse Brain
Authors: Tamai Yoichi , Kojima Hisako , Ohtani Yoshio , Uchida Kikue , Taguchi Fumiaki , Kawaguchi Takeshi , Miura Sadanori , Tateishi Jun
Published: November 2013

To determine the intracellular localization of the Creutzfeldt‐Jakob disease (CJD) agent in mouse brain, cerebrum tissue of the mouse brain affected with the Fukuoka‐1 strain was separated into six subcellular fractions (microsome, nerve ending, myelin, mitochondria, nucleus, and soluble fractions) by differential sucrose density gradient, and then the CJD infectivity of these fractions was examined. Serially diluted samples of each subfraction were inoculated intracerebrally into groups of BALB/c mice, and the infectivity was determined as to end point titration value, incubation period, and number of affected mice. On the basis of the protein content, the highest CJD infectivity was observed in the microsomal fraction. The nerve ending (synaptic plasma membrane) and myelin fractions were also infective. The mitochondria and nucleus fractions showed the lower infectivity. The infectivity of the soluble fraction was the lowest among the six subcellular fractions. From the findings obtained in this study two possibilities as to the intracellular localization of CJD agent were suggested: 1) the transmissible agent of CJD is closely associated with surface membranes of neuronal and/or glial cells, including their processes; 2) the CJD agent is diffusely present intracellularly, including in the surface membranes, but for manifestation of infectivity the agent needs membrane components as prerequisite factors.
Influence of Transglutaminase on the Functions of Mouse Peritoneal Macrophages
Authors: Ishitani Kouki , Suzuki Masuko
Published: November 2013

Influence of transglutaminase on the production of interleukin‐1 (IL‐1) and on the release of active oxygen from mouse peritoneal macrophages was examined using cystamine and methylamine, an enzyme inhibitor and a substrate inhibitor, respectively. Casein‐elicited or lipopolysaccharide (LPS)‐elicited macrophages have higher levels of transglutaminase activity in comparison with resident macrophages, and there exists a definite correlation between endocytosis of erythrocytes and transglutaminase activity in either group of macrophages. The release of IL‐1 by resident macrophages stimulated with LPS in vitro was significantly inhibited by the treatment with both transglutaminase inhibitors. However, these inhibitors were not able to inhibit the release of IL‐1 from casein‐elicited macrophages stimulated with LPS in vitro. The production of active oxygen from LPS‐elicited macrophages was inhibited in a dose‐dependent manner by the treatment of macrophages with cystamine, but was not by the treatment with methylamine. However, the treatment of LPS‐elicited macrophages with cystamine did not inhibit the uptake of glucose into macrophages. These results suggest that transglutaminase activity in mouse peritoneal macrophages is an important factor for macrophage functions.
Possibility of Bacteroides gingivalis Hemagglutinin Possessing Protease Activity Revealed by Inhibition Studies
Authors: Nishikata Makoto , Yoshimura Fuminobu , Nodasaka Yoshinobu
Published: November 2013

Inhibition of hemagglutinin (HA) activity in a membrane fraction of Bacteroides gingivalis was examined using various compounds. Leupeptin and antipain inhibited the HA activity at nm order. This potency was lost when the aldehyde group of leupeptin was converted to an alcohol moiety. Irreversible protease inhibitors, tosyl‐l‐lysine chloromethyl ketone (TLCK), p‐chloromercuribenzoate (PCMB), and N‐ethylmaleimide (NEM) were also inhibitory. From the inhibition experiments, we speculate that the HA possesses protease activity and that the same site of the molecule participates in the erythrocyte binding and the substrate binding.
Electron Microscopic Observations on Ciliated Epithelium of Tracheal Organ Cultures Infected with Bordetella bronchiseptica
Authors: Sekiya Kachiko , Futaesaku Yutaka , Nakase Yasukiyo
Published: November 2013

Using mouse tracheal organ cultures, the pathogenic effect of Bordetella bronchiseptica to epithelial cells was studied by electron microscopy. The ultrastructure of epithelial cells in uninfected tracheal rings was preserved well for longer than 3 days. In mouse tracheal rings infected with graded doses (3 × 105 to 107 CFU/ml) of phase I B. bronchiseptica, the colonization in the interciliary spaces of ciliated epithelial cells was observed after a 20‐hr infection period. The infected tracheal rings showed swelling of nonciliated cells as well as ciliated cells, rupture of cell membrane of cilia, swelling and disappearance of cilia, and atrophic cytomorphosis of epithelial cells. The severity of these changes occurred depending on the infection doses. These changes were essentially similar to those observed previously in the tracheal epithelia of the B. bronchiseptica‐infected mice. The usefulness of this in vitro model was suggested for studying the pathogenesis of Bordetella infection.
Cell Surface of Micrococcus luteus: Chemical Treatment of the Cells and Teichuronic Acids on the Surface
Authors: Monodane Toshihiko , Kusamichi Masako , Tokunaga Michiko , Torii Mitsuo
Published: November 2013

Micrococcus luteus IFO 3333 cells, 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. The surface of a M. luteus cell consisted of two or three areas with borders—the rough and the smooth areas, or the rough, the slightly rough, and the smooth areas; fluffy materials were clearly seen in the rough area. Gold particles were observed uniformly and densely on the whole cell surface. However, either mild acid treatment or mild Smith degradation of the cells altered the fluffy rough area to a rough one, and extremely decreased the agglutinability and the binding of protein A‐gold particles. Teichuronic acids appeared to be distributed uniformly on the whole cell surface of M. luteus IFO 3333.
Comparison of Virulence between Two Strains of Rattus Serotype Hemorrhagic Fever with Renal Syndrome (HFRS) Virus in Newborn Rats
Authors: Zhang Xian‐kui , Takashima Ikuo , Mori Fumiaki , Hashimoto Nobuo
Published: November 2013

Two strains of hemorrhagic fever with renal syndrome (HFRS) virus from Rattus, SR‐11 and KI‐262, showed virtually identical antigenicity but differed from prototype strain Hantaan 76–118 (Apodemus origin) in a neutralization test. Wistar newborn rats inoculated intraperitoneally (i.p.) with SR‐11, which was isolated from a laboratory rat associated with an outbreak of HFRS, developed clinical signs such as ataxia and limb paralysis and died at about 18 days after inoculation. The LD50 of SR‐11 in 1‐day‐old rats was 101.2 focus‐forming units (FFU). In contrast, the animals inoculated i.p. or intracerebrally with 104 FFU of KI‐262, which was from a wild rat in a dumping‐ground area—an enzootic focus where no human cases have been recorded—did not show any significant clinical signs. The susceptibility of rats to SR‐11 fatal infection was age‐dependent. Virus titers in brains, lungs, kidneys, and livers of the rats inoculated with SR‐11 were significantly higher than those in the same organs of the animals infected with KI‐262. Necrosis of neurons in the brain tissue occurred in the rats infected with SR‐11, while it was mild in the animals infected with KI‐262.
Isolation of Line 10 Hepatoma‐Cell Membrane by the Water Extraction Method and Immunochemical Analysis
Authors: Ahsan Chowdhury Rafiqul , Sasaki Jin‐ichi
Published: November 2013

Water extraction method was applied to isolate the cell membrane from line 10 hepatoma cells and normal liver cells in strain 2 guinea pig. The materials isolated by this method were further analyzed by different immunochemical techniques including SDS‐PAGE, crossed immunoelectrophoresis and crossed affino immunoelectrophoresis to demonstrate the major components and their antigenicities. Five major glycoproteins of apparent molecular weights of 44, 46, 62, 64, and 68 kDa were prominent in line 10 tumor cell materials, whereas one band of molecular weight of 82 kDa was prominent in the materials from normal liver cells. Also four minor components from line 10 tumor cells were found to be glycoprotein in nature.
Characterization of Activation of a Partially Defective B Cell Subpopulation with Immunocompetence Restricted to IgM and IgA
Authors: Nakaniwa Hiroshi , Kajikawa Taketsugu , Bito Yukio
Published: November 2013

A B‐cell subpopulation (BM‐A cell) responding to an antigen with the production of IgM and IgA plaque‐forming cells but not of IgG plaque‐forming cells was isolated from neonatally bursectomized chickens and was examined for the mode of activation by B‐cell mitogens. The BM‐A cells did not elevate both glucose consumption and protein synthesis with the B‐cell mitogens, in striking contrast to normal B cells. The BM‐A cells were also not activated by an activator of protein kinase C, phorbol myristate acetate. Both anti‐Ig and a calcium ionophore, A23187, however, primed the BM‐A cells to increase intracellular free calcium ion as well as normal B cells. From these results it is conceived that the lack of protein kinase C activation may be responsible for the failure of activation of BM‐A cells.
Induction of Resistance with Heat‐Killed Unencapsulated Strains of Staphylococcus epidermidis against Challenge with Encapsulated Strains of Staphylococcus epidermidis
Authors: Ichiman Yoshitoshi , Yamada Tetsuo , Yoshida Kosaku
Published: November 2013

Active immunization of mice with high doses of heat‐killed unencapsulated strains of Staphylococcus epidermidis, which were grown in brain heart infusion media, protected mice against challenge with encapsulated strains of S. epidermidis. The unencapsulated strains were capable of absorbing the protective antibody in rabbit hyperimmune sera prepared with the encapsulated strains. Also, mice treated with rabbit hyperimmune sera prepared with the unencapsulated strains were protected against challenge with the encapsulated strains. The protective activities of these rabbit hyperimmune sera were assumed to be essentially identical to those of the protective antibody induced by the encapsulated strains.
Characterization and Reassembly of a Regular Array in the Cell Wall of Clostridium difficile GAI 4131
Authors: Masuda Kuniyoshi , Itoh Masumi , Kawata Tomio
Published: November 2013

The cell wall of Clostridium difficile GAI 4131 was revealed by electron microscopy to have an outer layer composed of a nearly square array and contained the two major proteins with molecular weights of 38 kDa and 42 kDa. The properties and reassembly of the two major proteins into the regular array were investigated. When the isolated cell walls were treated with hydrophobic bond‐disrupting agents or a chelating agent specific for Ca2+, the two major proteins were effectively removed and the regularly arranged outer layer disappeared. The amino acid composition of the two major proteins differed from each other. The two major proteins also gave different peptide maps from each other upon proteolysis with Staphylococcus aureus V8 protease. The major proteins solubilized from the isolated cell walls with 8 m urea or 4 m guanidine hydrochloride could be reassembled into open‐ended cylinders possessing the native regular pattern by dialysis against neutral buffer containing 5 mn CaCl2. The reassembled cylinders purified by centrifugation on a Percoll density gradient were composed of almost equal amounts of the 38 kDa and 42 kDa proteins and freed from the other proteins. These results suggest that the regular array in the outer cell wall layer is constructed from the two major cell wall proteins and requires Ca2+ for its assembly.
Role of Pertussigen (Pertussis Toxin) on the Mouse Protective Activity of Vaccines Made from Bordetella Species
Authors: Munoz John J. , Peacock Marius G.
Published: November 2013

Pertussigen [pertussis toxin (Ptx)] from Bordetella pertussis, when detoxified, induces protection in mice to intracerebral challenge (ic) with virulent B. pertussis. In its native form, minute nonprotective doses promote the development of immunity induced by other antigens of B. pertussis. As little as 4 ng of Ptx, given with a nonprotective dose of 8 × 107 killed cells of the phase III Sakairi strain, promoted detectable protection to ic challenge. Native Ptx in doses of 0.4 to 400 ng did not protect mice, and vaccines made from strains not producing Ptx induced only weak protection. The marked enhancing action of Ptx was also observed with 5 μg of purified filamentous hemagglutinin and with vaccines made from other species of the Bordetella genus, such as B. parapertussis and B. bronchiseptica, but it was not observed with B. pertussis endotoxin. In addition, Ptx was still effective when given as late as 7 days after the vaccine. Antibodies to surface antigens of the challenge strain were demonstrated in sera of mice immunized with vaccines prepared with the different Bordetella species tested, but antibodies to Ptx were detected only in the sera of mice immunized with the wild‐type B. pertussis strains. Glutaraldehyde detoxified Ptx does not have this action. Pretreatment of normal mice with Ptx, also enhanced the protective action of a mouse antiserum to a wild‐type strain of B. pertussis. These observations show that antigens other than Ptx are responsible for the protection, and that Ptx acts non‐specifically to enhance the mouse protective action of those antigens.
Presence of Proteins Derived from the Vegetative Cell Membrane in the Dormant Spore Coat of Bacillus subtilis
Authors: Fujita Yuko , Yasuda Yoko , Kozuka Satoshi , Tochikubo Kunio
Published: November 2013

To confirm the presence of the outer spore membrane in dormant spore coats of Bacillus subtilis, the proteins from vegetative cell membrane and dormant spore coat fractions were compared by immunoblot assay with antibodies prepared against both preparations. The spore coat fraction contained at least 11 proteins antigenically identical to those in the vegetative cell membranes. Further, the cytochemical localization of the proteins derived from vegetative cell membrane in dormant spores was examined by an immunoelectron microscopy method with a colloidal gold‐immunoglobulin G complex. The colloidal gold particles were observed in the coat region and around the core region of dormant spore. These results have provided evidence that some proteins from vegetative cell membrane remain in the dormant spore coat region of B. subtilis, although it is not clear whether the outer membrane persists as an intact functional entity or not.
Characterization of a Strain of Mycoplasma hominis Lacking 120 kDa Membrane Protein Isolated from Vero Cell Culture
Authors: Sasaki Tsuguo , Sasaki Yuko , Matsumura Toshiharu , Oyama Noriko , Koshimizu Kaoru
Published: November 2013

A strain of Mycoplasma hominis lacking a major membrane protein of 120 kDa was isolated from a Vero cell culture. This strain showed very slow growth rate and formed nipple‐less colonies on agar medium.
Spreading of Streptomycin Antibiotic Resistance Gene in Escherichia coli Plasmids Demonstrated by Southern Blot Analysis
Authors: Hajnal Anna , Falus Andras
Published: November 2013

Plasmids from E. coli strains of 38 donors were transconjugated to common recipient SY663 Escherichia coli K12. The restriction patterns of the isolated plasmids were highly heterogenous. However, the streptomycin (Sm) resistance genes of the plasmids were identical or closely homologous in 29 of the 33 plasmids conferring Sm resistance. These data were based on Southern blot analysis, using the Sm resistance gene (encoding aminoglycoside phosphoryl transferase) as probe cut out from pBPl plasmid. Our data suggest an extensive spreading of streptomycin resistance gene of this type.
Characterization of a Nalidixic‐Acid‐Resistant Mutant of Escherichia coli as a Strict Aerobe
Authors: Yoshizawa Yukio , Yamamoto Nobuto
Published: November 2013

An Escherichia coli mutant, C18, which plates at an efficiency of 5.0 × 10−4 under anaerobic condition, was isolated among spontaneous nalidixic‐acid‐resistant mutants. This strict aerobic mutation was mapped by Pl cotransduction with a gyrA linked transposon Tn10 and found to be at the gyrA gene. A low degree of superhelicity of pBR322 DNA isolated from C18 was demonstrated by agarose gel electrophoresis with various concentrations of ethidium bromide. The superhelical density of pBR322 isolated from C18 was 80% of the value of pBR322 isolated from wild‐type bacteria cultured under aerobic condition, and 50% cultured under anaerobic condition. These results lead us to conclude that a certain mutation of the gyrA gene causes a decrease in DNA superhelicity and prevents anaerobic growth.
Evaluation of Production and Characterization of Monoclonal Antibodies to Human IgG of Four Subclasses
Authors: Harada Shigenori , Nishimura Shinji , Saiga Akihiko , Hosoi Susumu , Mikawa Haruki
Published: November 2013

Human IgG of four subclasses, semi‐purified from pooled human serum by a series of DEAE ion exchange and protein A affinity chromatographies, were used as immunogens and initial screening antigens to produce subclass‐specific and ‐restricted monoclonal antibodies (McAbs). These McAbs were bound to CNBr‐activated Sepharose 4B and utilized in immunoaffinity chromatography to prepare four polyclonal human IgG subclasses of satisfactory purities, which were then used as final screening antigens. Subclass‐specific McAbs thus chosen were further evaluated for subclass‐ and especially allotype‐specificity using a panel of monoclonal IgG myeloma proteins with representative Gm markers for each subclass in micro enzyme‐linked immunosorbent assay (ELISA). A total of 10 clones of subclass‐specific McAbs (one for anti‐IgG1, three anti‐IgG2, two anti‐IgG3, four anti‐IgG4) were established. Among them, IgG2‐specific clones of HG2‐30F and HG2‐56F, IgG3‐specific HG3‐7C and HG3‐32C, and IgG4‐specific HG4‐53G McAbs were superior to the corresponding specificity standard McAbs chosen by the Human Immunoglobulins Subcommittee of the WHO/International Union of Immunological Societies (IUIS) in 1985. As allotype‐specific McAbs, HG1‐1E for Glm(az) and HG3‐3B for G3m(b) were obtained. In micro ELISA of this study as well as all protocols of the previous WHO/IUIS collaborative study, antigens (myeloma IgG subclasses) were immobilized or fixed to a solid phase, resulting in possible variations in their epitope expressions. We developed a new assay system, micro radioimmunoassay (RIA), in which reactivities of McAbs against free IgG subclasses in solution can be evaluated. HG2‐30F, having extremely high reactivities to coated IgG2 in micro ELISA, remarkably reduced its reactivities to free IgG2 in solution in micro RIA. Two other clones also showed some different reactivities in micro RIA and micro ELISA. We believe that this micro RIA is valuable for evaluation of McAbs reactivities against native human IgG subclasses in solution.
Electron Microscopic Studies of Viruses Labeled with Magnetite
Authors: Mizutani Hiromichi , Mizutani Hiroko , Nozaki Kiyoe , Fujiwara Koichi
Published: November 2013

We were able to develop a method with which to successfully and specifically detect virus particles under the electron microscope by using magnetite. This method was devised on the principle that magnetite‐labeled antibody or magnetite coupled with protein A selectively bind virus or antibody‐treated virus particles on the electron microscope grid by the action of an electromagnet. Another advantage characterizing the technique is the possibility of detection of a small number of virus particles. This is done through a process of concentration and purification of the reaction complexes trapped rigidly by magnetic force.
Frequency of Cell Transformation by the Small DNA Tumor Viruses: Infection of Proliferating Cells and Quiescent Cells
Authors: Matsuzaki Akinobu , Okuda Atsuyuki , Tamura Hideaki , Ohtsu Masumi , Kimura Genki
Published: November 2013

Small DNA‐containing tumor viruses (simian virus 40, mouse polyomavirus, and adenoviruses) malignantly transform fibroblasts of the susceptible rodents. Fibroblasts can exist, in vitro and in vivo, in either of the two states: the proliferating state or the quiescent state. In the present study, we examined whether the state of fibroblasts at the time of exposure to these DNA viruses affects the frequency of transformation. Dense‐focus formation in monolayer culture of rat 3Y1 fibroblasts was used to quantitate transformation. Results show that the frequency of transformation by simian virus 40 and mouse polyomavirus was reduced when cells were in the proliferating state at the time of virus inoculation as compared to cells in the quiescent state, whereas that by adenovirus type 12 was similar in the two cellular states. The reduction of the frequency of transformation in proliferating cells infected with simian virus 40 was also observed in BALB/c 3T3 mouse cells. Mechanisms underlying the difference between the two cellular states and the difference between the papovavirus and adenovirus in this aspect of transformation remain to be investigated.
Immunopathology of Chronic Progressive Hepatitis in Nude Mice Infected with Low‐Virulent Mouse Hepatitis Virus
Authors: Midoro Katsuo , Nakanaga Kazue , Kyuwa Shigeru , Fujiwara Kôsaku
Published: November 2013

Progressive hepatitis in athymic nude (nu/nu) mice due to a low‐virulent mouse hepatitis virus, MHV‐2 cc, was examined for involvement of immunocytes and serum antibodies. At 3 to 6 weeks postinoculation (p.i.) a considerable number of Mac 1‐ and asialo GM1‐positive cells were accumulated in the affected liver and spleen. There were also some Thy‐1‐positive cells. Later than 2 weeks p.i., serum IgG and IgM antibodies were detected in parallel with virus‐neutralizing activity, while the IgG levels were lower than those of infected euthymic (nu/ +) littermates. By transfer of the infected nu/nu mouse serum, the recipient euthymic mice acquired resistance to lethal challenge infection with a virulent virus, MHV‐2.