bluuebunny/crossref_metadata_2025_split huggingface dataset:
Electron Microscopic Studies on Intracellular Multiplication of Rickettsia tsutsugamushi in L Cells
Authors: Urakami Hiroshi , Tsuruhara Takashi , Tamura Akira 
Published: November 2013

AbstractThe mechanism and kinetics of intracellular growth of Rickettsia tsutsugamushi were investigated by electron microscopic observations, parallel with quantitative analysis by counting the rickettsiae seen in electron micrographs and by plaque assay for infectivity of the culture. The observations demonstrated the existence of electron‐less dense and ‐dense types of rickettsiae in the early stage of infection, binary fission and the process of release of the microorganisms in the host cell cytoplasm and from the cell surface, formation of abnormally long rickettsiae, and the process of lysis of the host cell in the later stage of infection with vacuole formation between the inner and outer leaflets of the host cell nuclear membrane. Separate titrations of infectivity of the cells and the culture fluid showed a very slow increase in infectivity in the culture fluid compared with the intracellular titer, suggesting that the progeny rickettsiae stay in the cell or at the cell surface for a relatively long period. Doubling time of the rickettsia was found to be about 9 hr.
Passive Immunization of Mice with Monoclonal Antibodies to Glycoprotein gB of Herpes Simplex Virus
Authors: Kino Yoichiro , Eto Tatsuo , Ohtomo Nobuya , Hayashi Yoshinobu , Yamamoto Masahiro , Mori Ryoichi 
Published: November 2013

AbstractTo investigate the protective ability of monoclonal antibodies (MCAs) to viral glycoprotein in herpes simplex virus (HSV) infection, athymic nude mice were inoculated intracutaneously with HSV type‐1 (HSV‐1) in the midflank. Three hours after inoculation, one group of mice was passively immunized with one of a series of MCAs to glycoprotein gB of HSV‐1, and a control group of mice was given phosphate buffered saline alone. The control mice died within 16 days after infection, whereas the mice passively immunized with any of the MCA showed suppressed development of skin lesions. Three of six mice given MCA failed to develop any visible lesions and no HSV could be isolated from the lumbar dorsal root ganglia of these mice 60 days after the challenge. BALB/c mice were also protected from infection with HSV type 2 by passive immunization with MCA to HSV‐1 gB.
Possibility of Association and Activation of Glucose Dehydrogenase during Germination and Outgrowth of Bacillus subtilis Spores
Authors: Tochikubo Kunio , Yasuda Yoko 
Published: November 2013

AbstractBoth a salt‐dependent form and an active form of glucose dehydrogenase [EC 1.1.1.47] were isolated from germinated spores of Bacillus subtilis disrupted in deionized water and 100 mM phosphate buffer (pH 6.6), and most of the enzyme isolated from young vegetative cells was the active form regardless of the conditions for breakage by sonication. The molecular weight of the salt‐dependent form of the enzyme was about 55,000 and that of the active form was about 120,000. From the above results and the results on the glucose dehydrogenase extracted from resting spores disrupted in deionized water and 100 mM phosphate buffer (pH 6.6) reported in a previous paper, we propose that glucose dehydrogenase is present in resting spores as a monomeric form and is activated with association in vivo during germination and outgrowth.
Bioluminescent Assay as a Potential Method of Rapid Susceptibility Testing
Authors: Kouda Masakazu , Ouchi Yasuo , Takasaki Yuko , Maeda Takahiro , Matsuzaki Hiroko , Nakaya Rintaro 
Published: November 2013

AbstractA method of rapid susceptibility testing by bioluminescent assay was developed. Correlation between the 50% inhibition dose of antimicrobics for bacterial adenosine triphosphate measured by bioluminescent assay and the minimum inhibitory concentration obtained by the broth dilution method was satisfactory. In the bioluminescent assay the incubation time required was only 90 min.
Biosynthetic Processing of Human Interleukin‐2 Receptor (Tac Antigen)
Authors: Wano Yuji , Uchiyama Takashi , Yodoi Junji , Uchino Haruto 
Published: November 2013

AbstractBiosynthetic processing of the T‐cell surface receptor for interleukin‐2 was investigated in a cultured human T‐cell line MT‐1 by means of metabolic and cell surface radiolabeling followed by immunoprecipitation with a monoclonal anti‐receptor antibody (anti‐Tac) and analysis by one‐ and two‐dimensional polyacrylamide gel electrophoresis.The nascent precursor of the receptor (Mr = about 40,000, pI = 6.2–6.5) underwent a post‐translational modification giving rise to the mature receptor (IL‐2R; Mr = 60,000–65,000, pI = 4.2–4.7) within 2–4 hr. The post‐translational processing of IL‐2R caused a 20,000–25,000 increase in apparent molecular weight and a 2.0–2.5 acidic shift in the isoelectric point. The increase in molecular weight was attributable mainly to addition of sugar residues including glucosamine and galactose, and the charge shift to the addition of sialic acids. A carboxylic ionophore monensin completely blocked the maturation of IL‐2R at the mid‐stage of the processing. Fatty acid attachment appeared to comprise one of the steps of the post‐translational modification.Two‐dimensional analyses of IL‐2R biosynthesis enabled identification of the precursor of IL‐2R and its intermediate forms, from which it was partially possible to estimate reactions involved in the maturation of the precursor molecule.
Possible Involvement of Transglutaminase in Endocytosis and Antigen Presentation
Authors: Teshigawara Keisuke , Kannagi Reiji , Noro Nobuhiro , Masuda Tohru 
Published: November 2013

AbstractExperiments were carried out to determine as to whether or not internalization of antigen is necessary for subsequent antigen presentation by accessory cells using monoamines which are known as transglutaminase (TGase) inhibitors. It was found that endoeytosis for immune complexes via Fc receptors such as sheep erythrocytes coated with IgG class antibody (EA) was different from receptor‐independent endoeytosis for soluble protein such as horse radish peroxidase (HRP) in the sensitivity to monoamines; methylamine inhibited the receptor‐dependent endoeytosis of immune complexes at a concentration of over 20 mM and the receptor‐independent endoeytosis of HRP at 2 mM, while dansylcadaverine (DC) inhibited both at a concentration of 100 μM. It was noteworthy that antigen‐specific T cell proliferation to splenic adherent cells pulsed with DNP9,6‐ovalbumin (DNP9,6‐OVA) was blocked strongly by DC as well, but weakly by methylamine. These results suggest the possibility that antigen presentation requires internalization of antigen by a mechanism such as receptor‐dependent endoeytosis for the subsequent reexpression of antigen on membranes. Furthermore, it was confirmed that TGase activity is high in peritoneal exeudate and spleen adherent cells, both of which have accessory cell activities for lymphocytes, suggesting the possibility that TGase might be involved intimately in receptor‐dependent endoeytosis and subsequent antigen presentation.
Clostridium ramosum, an IgA Protease‐Producing Species and Its Ecology in the Human Intestinal Tract
Authors: Senda Shigeru , Fujiyama Yoshihide , Ushijima Tsutomu , Hodohara Keiko , Bamba Tadao , Hosoda Shiro , Kobayashi Kunihiko 
Published: November 2013

AbstractA bacterial strain isolated from feces of a patient with ulcerative colitis, which had been shown to produce a novel immunoglobulin A (IgA) protease (cleaving both the human IgA1 subclass and IgA2 subclass of A2m(1) allotype) extracellularly, was identified as Clostridium ramosum. By using a selective medium (proprion‐ate‐rifampicin‐gentamicin‐colimycin‐polymyxin medium) devised for C. ramosum, analysis of the population level of this organism was performed to determine its ecology in the human intestinal tract. C. ramosum was isolated in 20 of 25 fecal samples (80%) from patients with inflammatory bowel disease (I.B.D.) and in 112 of 135 samples (83%) from patients without I.B.D. (control group). C. ramosum was also isolated from 6 of 11 biopsy samples (55%) of the inflamed rectal mucosa from patients with ulcerative colitis and from five of 15 samples (33%) from the intact mucosa of the control group. The population levels of C. ramosum in most of the biopsy samples ranged from 2.3 to 5.0 log10 per gram. The IgA protease‐positive C. ramosum was found in only four of 135 fecal samples (3%) and one of 15 biopsy samples (6.7%) from the control group. These results indicate that IgA protease‐positive C. ramosum is not likely to play a role in the induction of I.B.D., unless the organism is first isolated from the patient with I.B.D.
Correlation of In Vitro Properties of Rhodococcus (Corynebacterium) equi with Virulence for Mice
Authors: Takai Shinji , Michizoe Toshitaka , Matsumura Kazuo , Nagai Motomi , Sato Hidefumi , Tsubaki Shiro 
Published: November 2013

AbstractTo study the virulence of Rhodococcus (Corynebacterium) equi, seven ATCC strains of different serotypes were tested for their LD50 in mice, clearance of the organism from the lungs and spleen following intravenous or intratracheal inoculation, and in vitro interaction with murine peritoneal macrophages. Strains ATCC 33704 and 33705 were virulent for mice and multiplied in the lungs and spleen, resulting in death of the animal in 5 days. The other five strains were avirulent for mice. The number of bacteria in the lungs and spleen of mice given these five strains decreased immediately. Pulmonary clearance of strains ATCC 33703, 33706, and 33707 was significantly more rapid than that of the virulent strains ATCC 33704 and 33705 12 hr after inoculation. Complete clearance of the avirulent strain ATCC 33707 occurred by day 14, while that of virulent ATCC 33704 and 33705 strains occurred by day 30. The virulent strains ATCC 33704 and 33705 were resistant not only to phagocytosis but also to intracellular killing by macrophages. Strains ATCC 33702 and 33706 were rapidly killed by macrophages although they were rather resistant to phagocytosis. Strain ATCC 33703 was easily phagocytized though resistant to killing by macrophages. The most avirulent strains, ATCC 33707 and 6939, were easily phagocytized and rapidly killed by macrophages. These results indicate that virulence appeared to be related to the ability of the organisms to resist clearance from the lungs and spleen and to resist phagocytosis and intracellular killing by macrophages.
Purification and Immunochemical Properties of a Protein Antigen from Serotype g Streptococcus mutans
Authors: Okahashi Nobuo , Koga Toshihiko , Hamada Shigeyuki 
Published: November 2013

AbstractA proteinaceous antigen (PAg) was purified from the culture supernatant of Streptococcus mutans 6715 (serotype g) by ultrafiltration, ammonium sulfate precipitation, DEAE‐Sephacel ion‐exchange chromatography, Phenyl‐Sepharose CL‐4B hydrophobic chromatography, and subsequent Sephacryl S‐300 gel filtration. A yield of 0.1 mg of PAg was obtained from a liter of culture supernatant. The isoelectric point and molecular weight of PAg were pH 4.6 and 210,000, respectively. It contained 35% sugar, which was identified as glucose by gas‐liquid chromatography. Amino acid analysis revealed that PAg contains 28% acidic and 11% basic amino acid residues. PAg retained its antigenicity after heating at 80 C for 10 min in deionized water, or after treatment with 0.1 M HCl or 0.1 M NaOH at 37 C for 1 hr. Immunodiffusion and Immunoelectrophoresis analyses revealed that PAg is serologically distinct from other cell‐surface antigens such as serotype‐specific polysaccharide and lipoteichoic acid. A cross‐reaction between PAg and a protein antigen similarly prepared from serotype c S. mutans was observed in immunodiffusion tests.
Mycobacterium fortuiturn Subspecies acetamidolyticum, a New Subspecies of Mycobacterium fortuiturn
Authors: Tsukamura Michio , Yano Ikuya , Imaeda Tamotsu 
Published: November 2013

AbstractMycobacterium fortuiturn subspecies acetamidolyticum is a new subspecies of M. fortuiturn and has an intermediate growth rate. It is a nonphotochromogenic mycobacterium. It does not utilize glutamate but utilizes acetamide as a simultaneous nitrogen and carbon source. It is able to utilize acetate, malate, pyruvate, fumarate, glucose, fructose, and n‐propanol as the sole sources of carbon in the presence of ammoniacal nitrogen, but does not utilize them in the presence of glutamate‐nitrogen. It is easily differentiated from all rapidly growing mycobacteria by its inability to utilize glutamate as a simultaneous nitrogen and carbon source, and from all slowly growing mycobacteria by its capacity to utilize acetamide as a simultaneous nitrogen and carbon source. Its mycolic acid pattern is different from that of M. fortuiturn. However, its deoxyribonucleic acid showed 94% relatedness with that of M. fortuiturn. In view of the above findings, it has been designated as a new subspecies of M. fortuiturn. The organism was isolated from sputum of a 56‐year‐old patient with lung disease and is considered to be a lung pathogen. The type strain is ATCC 35931 (NCH El 1620).
Purification of Scrapie Agent from Infected Animal Brains and Raising of Antibodies to the Purified Fraction
Authors: Takahashi Kaname , Goto Hitoshi , Shinagawa Morikazu , Doi Satoshi , Sato Gihei , Sasaki Satoshi 
Published: November 2013

AbstractThe fraction (P4) containing scrapie infectivity was obtained by treatment of scrapie‐infected mouse brains with the detergent sarcosyl, differential centrifugation, and proteolytic enzyme digestion. Scrapie infectivity in the P4 fraction was purified 239–2,390 times with respect to protein. Similar fractions were also prepared from the brain of a sheep naturally infected with scrapie. Morphological observation of the P4 fractions revealed that the main components were unique rods of 3–5 × 60–200 nm, which resembled scrapie‐associated fibrils (SAF) or prion rods. The P4 fractions formed three major broad bands of polypeptides with molecular weights (MWs) of about 24.5K, 21K, and 17K dalton (Kd) in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, and some low MW polypeptides were also present in the fraction. Rabbits immunized with this fraction prepared from mouse brains raised antibodies against the three major polypeptides.
Antichlamydial Activity of Ofloxacin
Authors: Hirai Katsuya , Une Tsutomu 
Published: November 2013

AbstractThe in vitro activity of ofloxacin, a new pyridone‐carboxylic acid, against 11 strains of Chlamydia trachomatis and six strains of Chlamydia psittaci was determined. All test strains of both species were inhibited by 0.39 μg of ofloxacin per ml. The antichlamydial activity of ofloxacin was comparable to that of doxycycline and four‐ to eightfold less than that of minocycline. The results of this susceptibility test, coupled with those of previous pharmacokinetic analyses of ofloxacin, warrant further evaluation of its clinical usefulness against chlamydial infections.
Microbial Adjuvant and Autoimmunity
Authors: Yokochi Takashi , Nakashima Izumi , Kato Nobuo , Asai Junpei , Miyadai Toshiaki , Kimura Yoshinobu 
Published: November 2013

AbstractThe role of humoral and cellular immune responses in the initiation and maintenance of autoimmune thyroiditis was investigated in mice immunized with syngeneic thyroid extract and Klebsiella O3 lipopolysaccharide (KO3 LPS) as an adjuvant. The transfer of spleen cells from hyperimmunized mice to 400R‐irradiated syngeneic mice produced definite lesions in the thyroid glands, whereas the transfer of immune sera failed to do so. No lesions were induced in normal intact mice by the same transfer of sera and spleen cells from hyperimmunized mice. It was suggested that the induction of thyroiditis by immunization using KO3 LPS adjuvant is primarily due to cell‐mediated immunity and that pretreatment of mice by X‐irradiation is essential for production of the lesions. The role of X‐irradiation in the induction of thyroiditis was discussed.
Dissection of the Role of Macrophages in Triggering T Lymphocytes for Interleukin 2 Production by Monoclonal Antibody OKT3
Authors: Fujimoto Kouji , Yamamoto Yusei , Ohmura Takafumi , Kawakami Kazuyoshi , Onoue Kaoru 
Published: November 2013

AbstractUnfractionated human peripheral blood mononuclear cells produce a small amount of interleukin 2 (IL 2) by stimulation with a monoclonal anti‐T3 antibody (OKT3) in vitro. The IL 2 production could be greatly augmented by the addition of a phorbol ester, 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA).In the presence of TPA, the T cell enriched fraction deprived of macrophages did not produce IL 2, but the T cells pulse‐incubated with OKT3 and reconstituted with macrophages efficiently produced IL 2 in subsequent culture in the presence of TPA as did T cells reconstituted with OKT3‐pulse‐incubated macrophages. The stimulating effect of OKT3 in the presence of macrophages was inhibited dose‐dependently by the addition of immunoglobulins, particularly by mouse IgG2a which is the same isotype as that of the OKT3 antibody, showing that it inhibits by blocking the binding of OKT3 to Fc receptors on macrophages. The same extent of IL 2 production was induced in T cells when paraformaldehyde‐fixed macrophages were substituted for intact macrophages. Remarkable IL 2 production was also induced by OKT3 when latex beads coated with rabbit anti‐mouse IgG2a antibody and TPA were added to the culture. It was confirmed that the production induced by these stimulations was due to an increase of IL 2 mRNA. These results show that effective signals for IL 2 production are generated by efficient crosslinking of T3 molecules which results from multi‐interaction of T3 molecules on the T cell membrane and anti‐T3 antibody molecules on macrophage membrane or on the surface of the latex particle.
Serum Antibodies to Individual Cytomegalovirus Structural Polypeptides in Renal Transplant Recipients during Viral Infection
Authors: Landini M.P. , Mirolo G. , Coppolecchia P. , Re M.C. , La Placa M. 
Published: November 2013

AbstractIn a longitudinal study we examined by immunoblotting (IB) the development and the evolution of the humoral immune response against individual cytomegalovirus (CMV) structural polypeptides in a total of 80 serum samples from 13 renal transplant recipients showing serological evidence of CMV infection and five renal transplant recipients with an anti‐CMV antibody level unchanged over the observation period. The results showed that the IB reactivity at the time of transplantation may be a good index of the host's humoral immune status against CMV; by using this procedure it is possible to identify a seroconversion by the detection of antibodies reacting with some intermediate molecular weight proteins in sera examined at high dilution. Furthermore, IB is a very sensitive procedure also for IgM detection as it anticipates the positivity of the enzyme immune assay for IgM.
Mechanism of Anti‐DNA Antibody Formation
Authors: Nakamura Kunie , Akahoshi Tohru , Kashiwazaki Sadao 
Published: November 2013

AbstractThe mechanism(s) of anti‐DNA antibody formation was comparatively investigated using in vitro human and murine B‐cell culture systems. T‐cell homogenate (TH) from SLE patients converted normal human B‐cells to anti‐DNA specific antibody‐forming plasma cells (anti‐DNA‐SPC) when cultured with calf thymic native DNA as antigen. TH of normal human donors suppressed the formation of anti‐DNA‐SPC from normal human B‐cell cultures even when SLE TH and DNA were added to the cultures. B‐cells derived from SLE patients were insensitive to normal human TH, and resulted in the formation of many anti‐DNA‐SPC. TH of young and old NZB mice stimulated the formation of anti‐DNA‐SPC from not only NZB but also C57BL/6 murine bone marrow cultures in the presence of DNA antigen. Human and murine TH, and both B‐cell cultures were reciprocally combined to test whether xenogeneic TH stimulated B‐cell cultures from different species. Xenogeneic TH was effective in triggering differentiation of xenogeneic B‐cells with respect to anti‐DNA‐SPC. The elimination of helper T‐subsets (Th) resulted in the generation of fewer anti‐DNA‐SPC, whereas the elimination of suppressor T‐subsets (Ts) caused the formation of many anti‐DNA‐SPC. Among organ homogenates, e.g., liver, kidney and, brain, and T‐cells from old NZB mice, TH was most effective in the stimulation of anti‐DNA‐SPC. The effective substance was sensitive to RNase‐A, but resistant to pronase and DNase‐I. Phenol extracted T‐cell RNA retained its activity. We concluded that the functional modulation of helper T‐cells, which reflects RNA molecules, could be the main etiology of autoantibody formation against DNA by both human and murine B‐cells.
Conjugal Transfer of Beta‐Lactamase‐Producing Plasmids of Neisseria gonorrhoeae to Neisseria meningitidis
Authors: Ikeda Fumiaki , Tsuji Akiyoshi , Kaneko Yasuko , Nishida Minoru , Goto Sachiko 
Published: November 2013

AbstractTwenty clinical isolates of beta‐lactamase‐producing Neisseria gonorrhoeae from Japanese sources were studied to define their ability to serve as donors for their plasmids in conjugation with Neisseria meningitidis. These twenty strains of N. gonorrhoeae harbored the 4.5‐megadalton (Mdal) beta‐lactamase‐producing plasmids and the 24.5‐Mdal conjugative plasmids. We found that only three of twenty N. gonorrhoeae strains showed a detectable conjugation frequency (>10‐5) with N. meningitidis as the recipient although all strains were capable of mobilizing beta‐lactamase‐producing plasmids to N. gonorrhoeae and to Escherichia coli. The 4.5‐Mdal beta‐lactamase‐producing plasmid was maintained in N. meningitidis, but the large 24.5‐Mdal conjugative plasmid has not been found in N. meningitidis transconjugants.
Role of L3T4 Antigen in the Activation of Various Functions of Lyt‐1+2‐ T Cells against Vaccinia Virus
Authors: Aoyama Atsuo , Yoshioka Takayuki , Sato Soichiro , Mizushima Yumiko , Ogata Masato , Ueda Shigeharu , Kato Shiro , Fujiwara Hiromi , Hamaoka Toshiyuki 
Published: November 2013

AbstractThe present study defines assay systems for vaccinia virus‐reactive Lyt‐1+2‐ T cells mediating various functions and investigates the positivity of L3T4 antigen on these Lyt‐1+2‐ T cells as well as the role of L3T4 antigen in the activation of these T cells with respect to their functions. C3H/He mice were immunized against vaccinia virus by inoculating viable virus intraperitoneally (i.p.). Antivaccinia virus reactivity in lymphoid cells from these immunized mice was assessed by a) proliferative response, b) helper T cell activities involved in cytotoxic T lymphocyte (CTL) and B cell (antibody) responses, c) delayed type‐hypersensitivity (DTH) response, and d) production of lymphokines such as interleukin 2 (IL2) and macrophage‐activating factor (MAF). The results demonstrate that all of the above anti‐vaccinia virus responses were mediated by Lyt‐1+2‐ T cells and that these Lyt‐1+2‐ T cells expressed L3T4 antigens on their cell surfaces. Moreover, such anti‐vaccinia Lyt‐1+2‐ T cell responses were inhibited in the presence of anti‐L3T4 antigen antibody. These results indicate that there is a reciprocal relationship between Lyt‐2 and L3T4 markers, and that L3T4 antigen is closely related to the activation of various functions of anti‐vaccinia virus Lyt‐1+2‐ T cells.